# Sticky  CCD/Neonicotinoid Data (Studies, Articles, Links)



## Barry

*Pathogen Webs in Collapsing Honey Bee Colonies*

Pathogen Webs in Collapsing Honey Bee Colonies


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## camero7

*Re: Pathogen Webs in Collapsing Honey Bee Colonies*

Temporal Analysis of the Honey Bee Microbiome Reveals Four Novel Viruses and Seasonal Prevalence of Known Viruses, Nosema, and Crithidia

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3110205/


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## Barry

*Effects of neonicotinoid insecticides on mammalian nicotinic acetylcholine receptors.*

Mammals and Neonics


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## camero7

J Econ Entomol. 2010 Oct;103(5):1517-23.

Weighing risk factors associated with bee colony collapse disorder by classification and regression tree analysis.

VanEngelsdorp D, Speybroeck N, Evans JD, Nguyen BK, Mullin C, Frazier M, Frazier J, Cox-Foster D, Chen Y, Tarpy DR, Haubruge E, Pettis JS, Saegerman C.
Source

Bureau of Plant Industry, Pennsylvania Department of Agriculture, 2301 North Cameron St., Harrisburg PA 17110, USA.
Abstract

Colony collapse disorder (CCD), a syndrome whose defining trait is the rapid loss of adult worker honey bees, Apis mellifera L., is thought to be responsible for a minority of the large overwintering losses experienced by U.S. beekeepers since the winter 2006-2007. Using the same data set developed to perform a monofactorial analysis (PloS ONE 4: e6481, 2009), we conducted a classification and regression tree (CART) analysis in an attempt to better understand the relative importance and interrelations among different risk variables in explaining CCD. Fifty-five exploratory variables were used to construct two CART models: one model with and one model without a cost of misclassifying a CCD-diagnosed colony as a non-CCD colony. The resulting model tree that permitted for misclassification had a sensitivity and specificity of 85 and 74%, respectively. Although factors measuring colony stress (e.g., adult bee physiological measures, such as fluctuating asymmetry or mass of head) were important discriminating values, six of the 19 variables having the greatest discriminatory value were pesticide levels in different hive matrices. Notably, coumaphos levels in brood (a miticide commonly used by beekeepers) had the highest discriminatory value and were highest in control (healthy) colonies. Our CART analysis provides evidence that CCD is probably the result of several factors acting in concert, making afflicted colonies more susceptible to disease. This analysis highlights several areas that warrant further attention, including the effect of sublethal pesticide exposure on pathogen prevalence and the role of variability in bee tolerance to pesticides on colony survivorship.

PMID:
21061948
[PubMed - indexed for MEDLINE]


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## camero7

Iridovirus and Microsporidian Linked to Honey Bee Colony Decline

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2950847/


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## squarepeg

great idea barry, thanks!

(all by randy oliver, and written from may 2012 - january 2013)

on ccd:

http://scientificbeekeeping.com/sick-bees-part-18a-colony-collaspse-revisited/

http://scientificbeekeeping.com/sick-bees-part-18b-colony-collapse-revisited/

http://scientificbeekeeping.com/sick-bees-part-18c-colony-collapse-revisited/

http://scientificbeekeeping.com/sick-bees-part-18d-colony-collapse-revisited/

http://scientificbeekeeping.com/sic...llapse-revisited-genetically-modified-plants/

http://scientificbeekeeping.com/sick-bees-part-18f-colony-collapse-revisited-pesticides/

on neonics:

http://scientificbeekeeping.com/neonicotinoids-trying-to-make-sense-of-the-science/

http://scientificbeekeeping.com/neonicotinoids-trying-to-make-sense-of-the-science-part-2/

http://scientificbeekeeping.com/a-new-large-scale-trial-of-clothianidin/


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## camero7

J Invertebr Pathol. 2012 Oct;111(2):106-10. doi: 10.1016/j.jip.2012.06.008. Epub 2012 Jul 20.

*Asymptomatic presence of Nosema spp. in Spanish commercial apiaries.*
Fernández JM, Puerta F, Cousinou M, Dios-Palomares R, Campano F, Redondo L.
Source

Apicultural Reference Center in Andalusia (CERA), Spain. [email protected]
Abstract

Nosemosis is caused by intracellular parasites (Nosema apis and Nosema ceranae) that infect the midgut epithelial cells in adult honey bees. Recent studies relate N. ceranae to Colony Collapse Disorder and there is some suggestion that Nosema spp., especially N. ceranae, induces high mortality in honey bees, a fact that is considered as a serious threat for colony survival. 604 samples of adult honey bees for Nosema spp. analysis were collected from beekeeping colonies across Spain and were analysed using PCR with capillary electrophoresis. We also monitored 77 Andalusian apiaries for 2 years; the sampled hives were standard healthy colonies, without any special disease symptoms. We found 100% presence of Nosema spp. in some locations, indicating that this parasite was widespread throughout the country. The two year monitoring indicated that 87% of the hives with Nosema spp. remained viable, with normal honey production and biological development during this period of time. The results of these trials indicated that both N. ceranae and N. apis could be present in these beehives without causing disease symptom and that there is no evidence for the replacement of N. apis by N. ceranae, supporting the hypothesis that nosemosis is not the main reason of the collapse and death of beehives.

Copyright © 2012 Elsevier Inc. All rights reserved.

PMID:
22820066
[PubMed - indexed for MEDLINE]


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## camero7

Pretty interesting paper on bees on sunflowers and soybeans in Uruguay. Really explains why bees do poorly on sunflowers treated with neonics. This is the one study I've found that really proves neonics hurt bees. But these hives were also hurt by organophosphates.

*Detection of Pesticides in Active and Depopulated Beehives in Uruguay*
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3210585/


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## camero7

*Colony Collapse Disorder in context*

Geoffrey R Williams,1,2,*,* David R Tarpy,3 Dennis vanEngelsdorp,4 Marie-Pierre Chauzat,5 Diana L Cox-Foster,4 Keith S Delaplane,6 Peter Neumann,7,8 Jeffery S Pettis,9 Richard E L Rogers,10 and Dave Shutler2

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3034041/


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## squarepeg

here is a definition of ccd from the usda/ars:

"The defining characteristic of CCD is the disappearance of most, if not all, of the adult honey bees in a colony, leaving behind honey and brood but no dead bee bodies. This definition has recently been revised to include low levels of Varroa mite and other pathogens, such as Nosema, as probable contributing factors. "

from "Colony Collapse Disorder: An Incomplete Puzzle" updated july 2012

http://www.ars.usda.gov/is/AR/archive/jul12/colony0712.htm


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## squarepeg

here's a good one:

http://www.nature.com/news/bees-lies-and-evidence-based-policy-1.12443

from the letter:

"As a scientist involved in this debate, I find this misinformation deeply frustrating. Yet I also see that lies and exaggeration on both sides are a necessary part of the democratic process to trigger rapid policy change. It is simply impossible to interest millions of members of the public, or the farming press, with carefully reasoned explanations. And politicians respond to public opinion much more readily than they respond to science."


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## camero7

Will banning neonicotinoids save pollinators?

Comments:

Lynn Dicks provides some background information and references to support her World View article, published this week in Nature.


On 25 February 2013, the European Union (EU)’s Standing Committee on the Food Chain and Animal Health votes on a proposal to ban the use of three widely used agricultural insecticides on crops attractive to bees. They are neonicotinoids - clothianidin, thiamethoxam and imidacloprid. They’ve been around since the mid-1980s. Often, they’re applied as seed treatments and remain in crop plant tissues throughout the plant’s life. This means their use is what you might call prophylactic, rather than being in response to pest attack.

The underlying driver for the sudden policy change is the emergence of new evidence showing substantial sublethal effects of neonicotinoids on honey bee Apis mellifera and bumblebee Bombus terrestris colonies at field-realistic doses [1,2,3]. These experiments imply particularly serious implications for wild bumblebee colonies, such as an 85% reduction in new queen production, if they are exposed in the wider environment at the levels tested [3].

Such effects are not considered acceptable risks by the European Food Safety Authority [4]. Even more importantly, they would not have been picked up by the existing regulatory system in Europe, which focuses entirely on honey bees, although new guidance to be published this spring is expected to extend the risk assessment to cover bumblebees and solitary bees.

We know that where measured, wild flower-visiting insects (bees, hoverflies, butterflies and moths) are declining in diversity and many are declining in abundance [5]. We know managed honey bees have suffered serious unexplained health problems leading to substantial colony losses in the US and parts of Europe [5]. And there is clear evidence that honey bees are exposed to neonicotinoids at levels similar to those tested, via crop plants, dust from seed planters and even weeds growing near treated fields, which seem to pick up residues from the soil [6].

The key question is, to what extent are the sublethal effects demonstrated in laboratories responsible for observed declines? This is where the scientific evidence flounders. Proving causal links between pesticide use and either bee declines or honey bee health problems is difficult. We still have no data on the actual exposure of wild pollinators to neonicotinoids, or to multiple pesticides including neonicotinoids, in their natural environment. The foraging behaviour and life histories of flower-feeding insects mean that reported levels of pesticide residue in crop plant nectar and pollen do not equate to actual exposure [7]. Most flower-feeding insects are generalists and opportunists. They feed on a range of available resources, including wild plants and crop plants. Landscape-scale field trials are needed, with treatment and control plots substantially larger than the standard 1 ha (100 m x 100 m), separated by a greater distance than the foraging range of bee colonies, which can be several kilometres. Such research is starting to happen. I know of at least one study recently commissioned in Sweden, but it will take years to come up with results. And so it should.

Current scientific opinion is that pollinator declines are caused by multiple interacting pressures rather than any single threat [5,8,9,10]. Habitat loss, disappearance of floral resources, climate change and disease may all play a part. Pesticides are one of these multiple, interacting pressures. There is no reason to believe that simply removing one group of insecticides, without addressing the other pressures, will solve the problem. It’s one step in the right direction.

Lynn Dicks is a Knowledge Exchange Fellow at the University of Cambridge, funded by the UK Natural Environment Research Council

Follow @LynnDicks on Twitter
1 Gill, R. J., Ramos-Rodriguez, O. & Raine, N. E. Combined pesticide exposure severely affects individual- and colony-level traits in bees. Nature 491, 105-108 (2012).
2 Henry, M. et al. A common pesticide decreases foraging success and survival in honey bees. Science 336, 348-350, doiOI 10.1126/science.1215039 (2012).
3 Whitehorn, P. R., O'Connor, S., Wackers, F. L. & Goulson, D. Neonicotinoid pesticide reduces bumble bee colony growth and queen production. Science 336, 351-352, doi:10.1126/science.1215025 (2012).
4 European Food Safety Authority. Conclusion on the peer review of the pesticide risk assessment for bees for the active substance clothianidin, imidacloprid and thiamethoxam. EFSA Journal 11, 3066-3068 (2013).
5 Potts, S. G. et al. Global pollinator declines: trends, impacts and drivers. Trends in Ecology & Evolution 25, 345-353, doiOI 10.1016/j.tree.2010.01.007 (2010).
6 Krupke, C. H., Hunt, G. J., Eitzer, B. D., Andino, G. & Given, K. Multiple Routes of Pesticide Exposure for Honey Bees Living Near Agricultural Fields. PLoS ONE 7, e29268 (2012).
7 Brittain, C. & Potts, S. G. The potential impacts of insecticides on the life-history traits of bees and the consequences for pollination. Basic Appl. Ecol. 12, 321-331, doiOI 10.1016/j.baae.2010.12.004 (2011).
8 Brown, M. J. F. & Paxton, R. J. The conservation of bees: a global perspective. Apidologie 40, 410-416, doiOI 10.1051/apido/2009019 (2009).
9 Szabo, N. D., Colla, S. R., Wagner, D. L., Gall, L. F. & Kerr, J. T. Do pathogen spillover, pesticide use, or habitat loss explain recent North American bumblebee declines? Conserv. Lett. 5, 232-239, doi:10.1111/j.1755-263X.2012.00234.x (2012).
10 Vanbergen, A. J. & Insect Pollinators Initiative. Threats to an ecosystem service: pressures on pollinators. Frontiers in Ecology and the Environment doi: 10.1890/120126 (In press).


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## Stromnessbees

*Effects of neonicotinoid dust from maize seed-dressing on honey bees *

Fabio S GOLASTRA 1, Teresa RENZI1, Stefano DRAGHETTI1, Piotr MEZYCKI2, Marco LODESANI, Stefano MAINI, Claudio PORRINI1
Dipartimento di Scienze Agrarie - Entomologia, Università di Bologna, Italy 
http://www.bulletinofinsectology.org/pdfarticles/vol65-2012-273-280sgolastra.pdf 
*
Abstract*

In Northern Italy from 2000 to 2008, many spring bee mortalities were clearly linked to sowing of maize seeds dressed with insecticides. In the present study, we investigated the effects on honey bees of clothianidin derived from maize seed-dressing (Poncho®) in laboratory (test by indirect contact) and in semi-field conditions. Despite the reduction of dust dispersion due to the application of the best available sowing techniques (pneumatic seeder equipped with deflector, improvement of seed-dressing quality) our results showed negative effect s on honey bees at individual level. In semi-field study, no effect was observed at the colony level despite the high bee mortality rate for 2-3 days after dust application. However, we can expect a colony decline and low honey production if this high forager mortality rate lasts for longer than 10 days. Such a situation is possible if the sowing period lasts several days as in the Po Valley, where the landscape is characterized by extended maize cultivation. Specific methodologies to assess the effects of dust have never been included in the official guidelines for the evaluation of side-effects of plant protection products on honey bees. For this reason, suitable and standardized methods for testing in laboratory and in semi-field conditions the effects on honey bees of contaminated dust dispersed during sowing were evaluated.


... This study is one of those that triggered the call for a ban of neonics in the EU.


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## Stromnessbees

DAFNE Project Nr. 100472
*MELISSA*

Investigations in the incidence of bee losses in corn and oilseed rape growing areas of Austria and possible correlations with bee diseases and the use of insecticidal plant protection products

Projektstart	01.03.2009
Projektende	15.03.2012

AuftragnehmerIn	Österreichische Agentur für Gesundheit und Ernährungssicherheit GmbH
WissenschaftlicheR ProjektleiterIn	Hofrat Dipl.-Ing. Leopold Girsch


Summing up, the results of the MELISSA-project give evidence that in Austria regional clustered bee damages had occurred in the years 2009 – 2011, which were *frequently associated with the use of maize and oilseed pumpkin seeds coated with insecticides, as proved by residue analysis*. The strong local component and the accumulation in areas with small-scale structured agriculture indicated special environmental conditions resulting in an increased exposition of honey bees to the identified insecticidal plant protection substances in the affected areas.
Regulatory measures to prevent honey bee losses due to the exposure of bees to insecticidal seed dressing substances have significantly improved the situation. However, repeatedly observed incidences of honey bee mortality in defined regions suggest their systematic correlation with local factors contributing to increased exposure of bees. In addition to considering environmental factors, all measures to mitigate risks have to be implemented invariably and with discipline.

http://www.dafne.at/dafne_plus_home...plus&content=result&come_from=&&search_fields[title_ger]=&search_fields[projektleiter]=&search_fields[antragsteller]=&search_fields[research_objective]=&search_fields[beauftragungsjahr]=&search_fields[offer_number]=100472&search_fields[keywords]=&search_fields[antragsteller_2]=&project_id=2909

Please enter the full title to your search engine to find this document, of go to www.dafne.at and search for document Nr.100472

English version half way down the page.


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## Stromnessbees

*In situ replication of honey bee colony collapse disorder
*
Chensheng LU 1, Kenneth M. W
ARCHOL 2, Richard A. CALLAHAN 3

Department of Environmental Health, Harvard School of
Public Health, Landmark Center West, Boston, MA, USA

Worcester County Beekeepers Association, Northbridge, MA, USA
Worcester County Beekeepers Association, Holden, MA, USA


*Abstract*

The concern of persistent loss of honey bee (Apis melliferaL.) colonies worldwide since 2006, a phenomenon referred to as colony collapse disorder (CCD), has led us to investigate the role of imidacloprid, one of the neonicotinoid insecticides, in the emergence of CCD. CCD is commonly characterized by the sudden disappearance of honey bees (specifically worker bees) from hives containing adequate food and various stages of brood in abandoned colonies that are not occupied by honey bees from other colonies. This in situ study was designed to replicate CCD based on a plausible mechanistic hypothesis in which the occurrence of CCD since 2006 was resulted from the presence of imidacloprid, one of the neonicotinoid insecticides, in high-fructose corn syrup (HFCS), fed to honey bees as an alternative to sucrose-based food. We used a replicated split-plot design consisting of 4 independent apiary sites. Each apiary consisted of 4 different imidacloprid-treated hives and a control hive. The dosages used in this study were determined to reflect imidacloprid residue levels reported in the environment previously. All hives had no diseases of symptoms of parasitism during the 13-week dosing regime, and were alive 12 weeks afterward. However, 15 of 16 imidacloprid treated hives (94%) were dead across 4 apiaries 23 weeks post imidacloprid dosing. *Dead hives were remarkably empty *except for stores of food and some pollen left, a resemblance of CCD. Data from this in situ study provide convincing evidence that exposure to sub-lethal levels of imidacloprid in HFCS causes honey bees to exhibit *symptoms consistent to CCD 23 weeks post imidacloprid dosing*. The survival of the control hives managed alongside with the pesticide-treated hives unequivocally augments this conclusion. The observed delayed mortality in honey bees caused by imidacloprid in HFCS is a novel and plausible mechanism for CCD, and should be validated in future studies. 

http://stream.loe.org/images/120406/Lu final proof.pdf


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## Stromnessbees

*Influence of dinotefuran and clothianidin on a bee colony *

Toshiro Yamada Kazuko Yamada Naoki Wada

Graduate School of Natural Science & Technology, Kanazawa University

http://moraybeedinosaurs.co.uk/neon...otefuran and clothianidin on a bee colony.pdf


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## Stromnessbees

*Multiple Routes of Pesticide Exposure for Honey Bees Living Near Agricultural Fields*

Christian H. Krupke,1,* Greg J. Hunt,1 Brian D. Eitzer,2 Gladys Andino,1 and Krispn Given1

*Abstract*

Populations of honey bees and other pollinators have declined worldwide in recent years. A variety of stressors have been implicated as potential causes, including agricultural pesticides. Neonicotinoid insecticides, which are widely used and highly toxic to honey bees, have been found in previous analyses of honey bee pollen and comb material. However, the routes of exposure have remained largely undefined. We used LC/MS-MS to analyze samples of honey bees, pollen stored in the hive and several potential exposure routes associated with plantings of neonicotinoid treated maize. Our results demonstrate that bees are exposed to these compounds and several other agricultural pesticides in several ways throughout the foraging period. During spring, extremely high levels of clothianidin and thiamethoxam were found in planter exhaust material produced during the planting of treated maize seed. We also found neonicotinoids in the soil of each field we sampled, including unplanted fields. *Plants visited by foraging bees (dandelions) growing near these fields were found to contain neonicotinoids as well*. This indicates deposition of neonicotinoids on the flowers, uptake by the root system, or both. Dead bees collected near hive entrances during the spring sampling period were found to contain clothianidin as well, although whether exposure was oral (consuming pollen) or by contact (soil/planter dust) is unclear. *We also detected the insecticide clothianidin in pollen collected by bees and stored in the hive*. When maize plants in our field reached anthesis, maize pollen from treated seed was found to contain clothianidin and other pesticides; and *honey bees in our study readily collected maize pollen*. These findings clarify some of the mechanisms by which honey bees may be exposed to agricultural pesticides throughout the growing season. These results have implications for a wide range of large-scale annual cropping systems that utilize neonicotinoid seed treatments.

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3250423/


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## Barry

You forgot this one on the same page:
*Exposure to clothianidin seed-treated canola has no long-term impact on honey bees.*

Cutler GC, Scott-Dupree CD.
Source

Department of Environmental Biology, Ontario Agricultural College, University of Guelph, Guelph, Ontario, Canada N1G 2W1. [email protected]
Abstract

We conducted a long-term investigation to ascertain effects on honey bee, Apis mellifera L., colonies during and after exposure to flowering canola, Brassica napus variety Hyola 420, grown from clothianidin-treated seed. Colonies were placed in the middle of 1-ha clothianidin seed-treated or control canola fields for 3 wk during bloom, and thereafter they were moved to a fall apiary. There were four treated and four control fields, and four colonies per field, giving 32 colonies total. Bee mortality, worker longevity, and brood development were regularly assessed in each colony for 130 d from initial exposure to canola. Samples of honey, beeswax, pollen, and nectar were regularly collected for 130 d, and the samples were analyzed for clothianidin residues by using high-performance liquid chromatography with tandem mass spectrometry detection. Overall, no differences in bee mortality, worker longevity, or brood development occurred between control and treatment groups throughout the study. Weight gains of and honey yields from colonies in treated fields were not significantly different from those in control fields. Although clothianidin residues were detected in honey, nectar, and pollen from colonies in clothianidin-treated fields, maximum concentrations detected were 8- to 22-fold below the reported no observable adverse effects concentration. Clothianidin residues were not detected in any beeswax sample. Assessment of overwintered colonies in spring found no differences in those originally exposed to treated or control canola. The results show that honey bee colonies will, in the long-term, be unaffected by exposure to clothianidin seed-treated canola.

http://www.ncbi.nlm.nih.gov/pubmed/17598537/


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## Stromnessbees

*Bee Deaths in France:* video 

http://youtu.be/9boueJGtLPY


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## Stromnessbees

*Honeybee Colony Collapse Disorder:* video from Japan

http://www.youtube.com/watch?v=BaqPNmk1kZY&list=PL3BEADE9DB257C5D1&index=30


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## Stromnessbees

*A Spring Without Bees *

by Michael Schacker

http://www.amazon.com/Spring-without-Bees-Collapse-Endangered/dp/B0058M8WSW#reader_B0058M8WSW

and here on video:

http://youtu.be/_ge8kFmujv0


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## Stromnessbees

*Honey bee survival rate better in west of Scotland *


> ...
> A further study led by Dr Connolly analysed colony failures over winter across the country. Of 89 colonies that had fed on oilseed rape, 27 failed, a death rate of 30 per cent. By contrast, 13 out of 82 colonies which had not fed on oilseed rape died – a smaller failure rate of 16 per cent.
> 
> Dr Connolly believes nicotine-based pesticides, neonicotinoids, may be contributing to the deaths of bees feeding on the crop, which is more commonly grown in the east.
> 
> He said: “All oilseed rape is treated with neonicotinoids, you can’t buy it without it being pre-treated with neonicotinoids.”
> ...


http://www.scotsman.com/news/enviro...val-rate-better-in-west-of-scotland-1-2807966


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## Stromnessbees

*Comparative Sublethal Toxicity of Nine Pesticides on Olfactory Learning Performances of the Honeybee Apis mellifera*

A. Decourtye, 1J. Devillers,2E. Genecque,3K. Le Menach,4H. Budzinski,4S. Cluzeau,1M. H. Pham-Del(gue3

*Abstract.*

Using a conditioned proboscis extension response (PER) assay, honeybees (Apis mellifera L.) can be trained to associate an odor stimulus with a sucrose reward. Previous studies have shown that observations of conditioned PER were of interest for assessing the behavioral effects of pesticides on the honeybee. In the present study, the effects of sublethal concentrations of nine pesticides on learning performances of worker bees subjectedto the PER assay were estimatedand compared. Pesticides were tested at three concentrations. The highest concentration of each pesticide corresponded to the median lethal dose value (48-h oral LD50), received per bee and per day, divided by 20. Reduced learning performances were observedfor bees surviving treatment with fipronil, deltamethrin, endosulfan, and prochloraz. A lack of behavioral effects after treatment with k-cyalothrin, cypermethrin, s-fluvalinate, triazamate, and dimethoate was recorded. No-ob-served-effect concentrations (NOECs) for the conditioned PER were derived for the studied pesticides. Our study shows that the PER assay can be usedfor estimating sublethal effects of pesticides on bees. Furthermore, comparisons of sensitivity as well as the estimation of NOECs, useful for regulatory purposes, are possible.

http://www.environmentalexpert.com/Files\6063\articles\4909\QM245Q254G1T6X0R.pdf


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## j.kuder

http://www.extension.org/pages/65034/neonicotinoid-seed-treatments-and-honey-bee-health


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## Stromnessbees

*Quantification of Imidacloprid Uptake in Maize Crops*

J. M. Bonmatin ,*† P. A. Marchand ,† R. Charvet ,† I. Moineau ,† E. R. Bengsch ,† and M. E. Colin ‡
Centre de biophysique moléculaire, CNRS (Centre National de la Recherche Scientifique) & Université d'Orléans, 45071 Orléans Cedex 02, France, and Laboratoire de Pathologie Comparée des Invertébrés, Université de Montpellier II, 34095 Montpellier Cedex 5, France
J. Agric. Food Chem., 2005, 53 (13), pp 5336–5341
DOI: 10.1021/jf0479362

Publication Date (Web): June 2, 2005

*Abstract*

The systemic imidacloprid is one of the most used insecticides in the world for field and horticultural crops. This neurotoxicant is often used as seed-dressing, especially for maize, sunflower, and rape. Using a LC/MS/MS technique (LOQ = 1 μg/kg and LOD = 0.1 μg/kg), the presence of imidacloprid has been measured in maize from field samples at the time of pollen shed, from less than 0.1 μg/kg up to 33.6 μg/kg. Numerous random samples were collected throughout France from 2000 to 2003. *The average levels of imidacloprid measured are 4.1 μg/kg in stems and leaves, 6.6 μg/kg in male flowers (panicles), and 2.1 μg/kg in pollen.* These values are similar to those found previously in sunflower and rape. These results permit evaluation of the risk to honeybees by using the PEC/PNEC ratios (probable exposition concentrations/predicted no effect concentration). PEC/PNEC risk ratios were determined and ranged between 500 and 600 for honeybees foraging on maize treated with imidacloprid by seed dressing. *Such a high risk factor can be related to one of the main causes of honeybee colony losses.*

http://pubs.acs.org/doi/abs/10.1021/jf0479362


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## Stromnessbees

*The corn pollen as a food source for honeybees*

http://www.scielo.br/pdf/asagr/v33n4/20.pdf 

DOI: 10.4025/actasciagron.v33i4.10533
Acta Scientiarum. Agronomy Maringá, v. 33, n. 4, p. 701-704, 2011

Darclet Teresinha Malerbo-Souza

*ABSTRACT*

This experiment was conducted on the campus of the University Center Moura Lacerda, Ribeirão Preto, in 2009 an d 2010 with the objective of verifying the attractiveness of corn pollen for the Africanized honeybees Apis mellifera. The frequency of these bees, and the foraging behavior and stability were obtained by counting the first 10 min. of each hour, from 7.00 to 18.00, on three different days. Africanized honey bees collected only pollen in male flowers of maize, with a peak frequency of 9.00 in 2009 and between 16.00 and 17.00 in 2010. The corn pollen was very collected by Africanized honey bees, *an important food source for these bees.*


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## Stromnessbees

*Evaluation of the nutritive value of maize for honey bees*

Nicole Höcherl
Reinhold Siede
Ingrid Illies
Heike Gätschenberger
Jürgen Tautz
a
BEEGroup, Biozentrum Universität Würzburg, Am Hubland, D-97074 Würzburg, Germany
b
LLH Bieneninstitut Kirchhain, Erlenstraße 9, D-35274 Kirchhain, Germany
c
LWG, Fachzentrum Bienen, An der Steige 15, D-97209 Veitshöchheim, Germany

Received 23 September 2011
Received in revised form 1 December 2011
Accepted 2 December 2011
Available online 7 December 2011

*abstract*

In modern managed agro-ecosystems, the supply of adequate food from blooming crops is limited to brief periods. During periods of pollen deficiencies, bees are forced to forage on alternative crops, such as maize. However, pollen of maize is believed to be a minor food source for bees as it is thought to be lack ing in proteins and essential amino acids. This study was conducted to verify this assumption. In maize, a strikingly low concentration of histidine was found, but the amount of all other essential amino acids was greater than that of mixed pollen. The performance and the immunocompetence of bees consuming a pure maize pollen diet (A) was compared to bees feeding on a polyfloral pollen diet (B) and to bees feeding on an artificial substitute of pollen (C). Consumption of diets A and C were linked to a reduction in brood rearing and lifespan. However, no immunological effects were observed based on two parameters
of the humoral immunity.

http://www.hobos.de/fileadmin/Publikationen/145_Hoecherl....pdf 



> ...
> Mixed pollen was collected by bees in June 2009 during the offbloom period of maize using commercial pollen traps. The pollen loads were removed daily in the evening and frozen to 18 C. Before the pollen was fed to the bees (colonies and caged bees) the pollen loads were ground and later mixed with honeydew honey (fir tree) to create a paste (ratio 2.5:1, wt/wt). *Maize pollen (variety ‘‘Athletico’’ KWS, Einbeck, Germany)* was collected by hand, ...


----------



## squarepeg

*eu vote*

http://www.hortweek.com/Retail/arti...ched-todays-meeting-eu-member-states-experts/


----------



## squarepeg

*Re: eu vote*

http://euobserver.com/environment/119446


----------



## squarepeg

*Re: eu vote*

the latest:

http://www.americanbeejournal.com/site/epage/81673_828.htm


----------



## squarepeg

*Re: eu vote*

peter l. borst writes on bee-l:

"What really happened:

Several European countries have suspended the use of certain pesticides in response to incidents involving acute poisoning of honey bees. To EPA's knowledge, none of the incidents that led to suspensions have been associated with Colony Collapse Disorder. The following are the countries in which pesticides have been suspended, the pesticides in question, and the current registration status for the pesticide:

France - Sunflower and corn seed treatments of the active ingredient imidacloprid are suspended in France; other imidacloprid seed treatments, such as for sugar beets and cereals, are allowed, as are foliar uses.

Germany - The use of a number of seed treatment pesticides was temporarily suspended following an incident in May 2008 in which many bees were inadvertently poisoned. However, after investigating the factors contributing to the situation, Germany lifted the suspensions with the exception of the neonicotinoid clothianidin, which remains suspended as a seed treatment for corn.

Italy - Certain imidacloprid and other neonicotinoid seed treatment uses were suspended temporarily, but foliar uses are allowed. This action was taken based on preliminary monitoring studies in northern and southern regions of Italy showing that bee losses were correlated with the application of seeds treated with these compounds; Italy also based its decision on the known acute toxicity of these compounds to pollinators.

Slovenia - Neonicotinoid seed treatments for maize and oil seed rape (canola) were temporarily suspended. The suspension was based on poor seed treatment methods resulting in release of dust during the seed sowing process. In August 2008, the suspension for oil seed rape seed treatments was lifted due to improved seed treatment methods and seed sowing equipment.

SOURCE:
http://www.epa.gov/pesticides/about/intheworks/ccd-european-ban.html "


----------



## squarepeg

*Re: eu vote*

for those who haven't seen it yet:

http://gallery.mailchimp.com/5fd2b1..._Happened_to_the_Bees_This_Spring2013_opt.pdf


----------



## borderbeeman

Central Archive for all Recent Papers on Neonics, Bee-Deaths and Bird Deaths associated with Neonics

http://smallbluemarble.org.uk/research/

Here is a sample:

http://smallbluemarble.org.uk/research/existing-scientific-evidence-of-neonicotinoids-bees/
This policy note from the EU Department for Economic and Scientific Policy contains some useful summaries of relevant research, including soil persistence rates, and defers in relation to pesticide authorisations to the on-going review by the European Food Safety Authority. The policy concludes that “ the precautionary principle in accordance with the Regulation (EC) No 1107/2009 should be applied when using neonicotinoids.”

http://smallbluemarble.org.uk/wp-content/uploads/2011/11/Mineau-Palmer-2013-The-Impact-of-the-Nations-Most-Widely-Used-Insecticides-on-Birds.pdf
The Impact of the Nation’s Most Widely Used Insecticides on Birds – American Bird Conservancy, March 2013. This report shows that similar errors in risk assessment have been made with regard to birds as were made to insects – the chronic/reproductive toxicity of neonicotinoids to birds is high. This was recognised very early on in the regulatory reviews of the various active ingredients. Yet high reproductive toxicity in birds is typically ignored in the pesticide review process.

http://smallbluemarble.org.uk/wp-content/uploads/2013/03/Hatjina2013.pdf
Sublethal doses of imidacloprid decreased size of hypopharyngeal glands and respiratory rhythm of honeybees in vivo


----------



## BlueDiamond

borderbeeman said:


> Central Archive for all Recent Papers on Neonics, Bee-Deaths and Bird Deaths associated with Neonics


If they wanted to, Bayer and Syngenta could hire a professional film crew and ecologists to make a pro-grade film documenting:

1) No food crops in the USA have suffered yield losses due to a shortage of honeybees.

2) Wildflowers growing in the vicinity of neonic treated crops in the USA continue to set alot of seed because both honeybees and wild pollinators like hover flies and butterflies continue to be common along the edges of the neonic treated crop fields.

3) Bats continue to be common in the evening in the vicinity of farm buildings that are surrounded by neonic treated crops because there are still lots of insects flying around in the evening.

4) Leopard frogs continue to be common along the margins of fields of Roundup Ready Corn and Soybeans in the Midwest USA.

5) Birds continue to be common in the tree hedgerows and shelter belts that are surrounded by neonic treated crops because there still lots of earthworms and other invertebrates for them to eat in the neonic treated crop soil plus they can find wildflower seeds along the crop margins.


----------



## borderbeeman

bluediamond said:


> if they wanted to, bayer and syngenta could hire a professional film crew and ecologists to make a pro-grade film documenting:
> 
> 1) no food crops in the usa have suffered yield losses due to a shortage of honeybees.
> 
> 2) wildflowers growing in the vicinity of neonic treated crops in the usa continue to set alot of seed because both honeybees and wild pollinators like hover flies and butterflies continue to be common along the edges of the neonic treated crop fields.
> 
> 3) bats continue to be common in the evening in the vicinity of farm buildings that are surrounded by neonic treated crops because there are still lots of insects flying around in the evening.
> 
> 4) leopard frogs continue to be common along the margins of fields of roundup ready corn and soybeans in the midwest usa.
> 
> 5) birds continue to be common in the tree hedgerows and shelter belts that are surrounded by neonic treated crops because there still lots of earthworms and other invertebrates for them to eat in the neonic treated crop soil plus they can find wildflower seeds along the crop margins.


please see moderator's introduction to this section, it is *not for discussion or debate *- only links to research papers


----------



## bearkarting

One of my co-worker received the following in an email.

FOR IMMEDIATE RELEASE
May 2, 2013

USDA and EPA Release New Report on Honey Bee Health


WASHINGTON -- The U.S. Department of Agriculture (USDA) and the U.S. Environmental Protection Agency (EPA) today released a comprehensive scientific report on honey bee health. The report states that there are multiple factors playing a role in honey bee colony declines, including parasites and disease, genetics, poor nutrition and pesticide exposure. 

"There is an important link between the health of American agriculture and the health of our honeybees for our country's long term agricultural productivity," said Agriculture Deputy Secretary Kathleen Merrigan. "The forces impacting honeybee health are complex and USDA, our research partners, and key stakeholders will be engaged in addressing this challenge."

"The decline in honey bee health is a complex problem caused by a combination of stressors, and at EPA we are committed to continuing our work with USDA, researchers, beekeepers, growers and the public to address this challenge," said Acting EPA Administrator Bob Perciasepe. "The report we've released today is the product of unprecedented collaboration, and our work in concert must continue. As the report makes clear, we've made significant progress, but there is still much work to be done to protect the honey bee population."


In October 2012, a National Stakeholders Conference on Honey Bee Health, led by federal researchers and managers, along with Pennsylvania State University, was convened to synthesize the current state of knowledge regarding the primary factors that scientists believe have the greatest impact on managed bee health. 

Key findings include:

Parasites and Disease Present Risks to Honey Bees:

The parasitic Varroa mite is recognized as the major factor underlying colony loss in the U.S. and other countries. There is widespread resistance to the chemicals beekeepers use to control mites within the hive. New virus species have been found in the U.S. and several of these have been associated with Colony Collapse Disorder (CCD).
Increased Genetic Diversity is Needed:

U.S. honeybee colonies need increased genetic diversity. Genetic variation improves bees thermoregulation (the ability to keep body temperature steady even if the surrounding environment is different), disease resistance and worker productivity. 
Honey bee breeding should emphasize traits such as hygienic behavior that confer improved resistance to Varroa mites and diseases (such as American foulbrood). 
Poor Nutrition Among Honey Bee Colonies:

Nutrition has a major impact on individual bee and colony longevity. A nutrition-poor diet can make bees more susceptible to harm from disease and parasites. Bees need better forage and a variety of plants to support colony health.
Federal and state partners should consider actions affecting land management to maximize available nutritional forage to promote and enhance good bee health and to protect bees by keeping them away from pesticide-treated fields.
There is a Need for Improved Collaboration and Information Sharing:

Best Management Practices associated with bees and pesticide use, exist, but are not widely or systematically followed by members of the crop-producing industry. There is a need for informed and coordinated communication between growers and beekeepers and effective collaboration between stakeholders on practices to protect bees from pesticides. 
Beekeepers emphasized the need for accurate and timely bee kill incident reporting, monitoring, and enforcement.
Additional Research is Needed to Determine Risks Presented by Pesticides:

The most pressing pesticide research questions relate to determining actual pesticide exposures and effects of pesticides to bees in the field and the potential for impacts on bee health and productivity of whole honey bee colonies.

Those involved in developing the report include USDA's Office of Pest Management Policy (OPMP), National Institute of Food and Agriculture (NIFA), Agricultural Research Services (ARS), Animal and Plant Health Inspection Service (APHIS), National Resource Conversation Service (NRCS) as well as the EPA and Pennsylvania State University. The report will provide important input to the Colony Collapse Disorder Steering Committee, led by the USDA, EPA and the National Agricultural Statistics Service (NASS).

An estimated one-third of all food and beverages are made possible by pollination, mainly by honey bees. In the United States, pollination contributes to crop production worth $20-30 billion in agricultural production annually. A decline in managed bee colonies puts great pressure on the sectors of agriculture reliant on commercial pollination services. This is evident from reports of shortages of bees available for the pollination of many crops.

The Colony Collapse Steering Committee was formed in response to a sudden and widespread disappearance of adult honey bees from beehives, which first occurred in 2006. The Committee will consider the report's recommendations and update the CCD Action Plan which will outline major priorities to be addressed in the next 5-10 years and serve as a reference document for policy makers, legislators and the public and will help coordinate the federal strategy in response to honey bee losses. 

To view the report, which represents the consensus of the scientific community studying honey bees, please visit: http://www.usda.gov/documents/ReportHoneyBeeHealth.pdf


----------



## Stromnessbees

*Neonicotinoids pose serious risk to aquatic ecosystems*

*Macro-Invertebrate Decline in Surface Water Polluted with Imidacloprid *

Tessa C. Van Dijk, Marja A. Van Staalduinen, Jeroen P. Van der Sluij

http://www.plosone.org/article/info:doi/10.1371/journal.pone.0062374 


Abstract

Imidacloprid is one of the most widely used insecticides in the world. Its concentration in surface water exceeds the water quality norms in many parts of the Netherlands. *Several studies have demonstrated harmful effects of this neonicotinoid to a wide range of non-target species.* Therefore we expected that surface water pollution with imidacloprid would negatively impact aquatic ecosystems. 

Availability of extensive monitoring data on the abundance of aquatic macro-invertebrate species, and on imidacloprid concentrations in surface water in the Netherlands enabled us to test this hypothesis. Our regression analysis showed a significant negative relationship (P<0.001) between macro-invertebrate abundance and imidacloprid concentration for all species pooled. A significant negative relationship was also found for the orders Amphipoda, Basommatophora, Diptera, Ephemeroptera and Isopoda, and for several species separately. The order Odonata had a negative relationship very close to the significance threshold of 0.05 (P = 0.051). However, in accordance with previous research, a positive relationship was found for the order Actinedida. 

We used the monitoring field data to test whether the existing three water quality norms for imidacloprid in the Netherlands are protective in real conditions. Our data show that macrofauna abundance drops sharply between 13 and 67 ng l−1. For aquatic ecosystem protection, two of the norms are not protective at all while the strictest norm of 13 ng l−1 (MTR) seems somewhat protective. In addition to the existing experimental evidence on the negative effects of imidacloprid on invertebrate life, our study, based on data from large-scale field monitoring during multiple years, shows that* serious concern about the far-reaching consequences of the abundant use of imidacloprid for aquatic ecosystems is justified.*


----------



## borderbeeman

*Is the solution to bees survival to ban more pesticides?*

http://www.sciencedaily.com/releases/2013/05/130503094140.htm










May 3, 2013 —
*Bees Survival: Ban More Pesticides?*
*Neonicotinoids are under intense scrutiny. But a ban on a broad variety of pesticides may be required to protect bees, humans and the environment.*

The European Commission, on 29th April 2013, slapped a two-year ban on insecticides suspected of killing-off bee colonies. This follows the European Food Safety Authority finding that they pose a high acute risk to honey bees. Studies suggest that the nicotine-like compounds fry bees' navigation systems and leave them unable to learn, while weakening their immune system.

But scientists now warn that other nerve-agents targeting insect pests may also be harming bees and other pollinators.

_*"These neonicotinoids are just one of hundreds of compounds being used and I would be surprised if it was all down to just these chemicals,"*_ says Christopher Connolly, a neuro-scientist at the University of Dundee, UK. He argues that we should not allow farmers spray a toxic soup of chemicals onto their crops.

* Pesticides not adequately tested*

Connolly exposed bee brains to these pesticides and organo-based pesticides and reported that the nerves spun into hyperactivity and then stopped working. A combination of these two pesticides types had a stronger impact, suggesting the combined soup of pesticides could be causing more serious harm.

_*"I don't understand how this was missed. As a neuroscientist it just seemed blindingly obvious. The biggest effect was hyper-activation of the major learning centre, which was completely predictable,"* _Connolly said.

The nerve agents effects were missed because safety-screens only looked to see how many honey bees die after four days exposure to the pesticide in question. But harm to the bees is only evident over a period of two weeks in bumblebees and is only seen when you look at entire colonies.
_*

"So the safety test is all wrong. The thing that concerns me is that this throws a question mark over several hundred pesticides, all tested by inadequate safety screens," * _says Connolly. He suggests that we should be tracking pesticides use in the environment, just like we monitor drug use in patients.

Not collecting such data might even pose health issues for people.
_*
"Bear in mind we have lots of 'idiopathic' diseases (diseases of unknown origin) in humans, which we don't know the cause of and given that we don't know what pesticides are used in what combinations and when, we don't know if these pesticides may be contributing to some or even all these unknown diseases,"*_ Connolly warns.

*More research needed*

Connolly argues that we need to carry out research to find out which pesticides are the least harmful. 
If neonicotinoids are the least toxic, then we should go with them. He says governments have under-funded this research area partly because it is inconvenient to find pesticides are dangerous. 

Dave Goulson, Professor of Biological Science at the University of Stirling, UK agrees:

_* "there haven't been nearly enough studies of all pesticides or interactions between them." *_ 
He recently published a study showing neonicotinoids hit bumblebee colony growth and queen production. 
He also said: _*"beneficial insects such as ladybirds and bees are exposed to lots of different chemicals and we have a really poor understanding of what it does to them."*_ He also points out that we need to be concerned with what we replace these nerve agents with.

More research may be helpful, but industry criticises extrapolation of lab studies to field conditions. Julian Little, spokesperson for Bayer Cropscience, based in Norwich, UK, says the evidence against these pesticides has all been lab based, essentially taking a social insect and force-feeding it insecticide. It says the results cannot be replicated in the environment.

But he also agrees more monitoring of pollinators is needed. _*"Where you do get large-scale bee deaths not enough has been done to know exactly what has happened,"*_ Little commented. He says pests and loss of feeding sites and nesting sites are most likely behind bee declines. _*"France has had restrictions [of neonicotinoids] over the last ten years, yet the bees there remain as bad if not worse than they are in the UK."*_Avoidance of pesticide use

A possible solution to preserve bee populations further would be to restore the principle of avoidance of pesticide use.

_*"The whole ethos of pest management has gone in the wrong direction," Goulson argues. Whereas integrated pest management sought to use as few pesticides as possible, the neonicotinoids are a preventive strike. 

"A simple analogy is that it's like taking antibiotics in case you get ill rather than when you get ill. Everyone knows that is a silly idea, as it results in bacteria rapidly developing resistance. It is the same with these pesticides."*_​
However, opponents believe that the neonicotinoids ban is unlikely to decrease pesticide use. Quite the opposite.
Julian Little of Bayer warns that farmers may now have to resort to spraying insecticides up to four times a year, now that they cannot coat seeds in neonicotinoids.

But other experts do not agree. There are several alternatives to using neonicotinoids, and other pesticides, according to Simon Potts, professor of biodiversity and ecosystem services at Reading University, UK.
_* "This is a great opportunity for farmers to adopt these practices to protect bees and other pollinators".*_Indeed, he believes farmers will benefit from healthy pollinator populations as they provide substantial economic benefits to crop pollination.

_* "Few people would disagree that we need to protect our food production, but it shouldn't be at the cost of damaging the environment," Potts said, adding:
"A short-term decision to keep using harmful products may be convenient, but will almost certainly have much greater long-term costs for food production and the environment."*_​


----------



## borderbeeman

*"canada wrestles with bee-killing pesticides"*

http://www.cbc.ca/news/canada/edmonton/story/2013/05/03/science-bee-neonicotinoid-pesticides.html

*CBC Edmonton: May 3, 2013 2:18 PM MT*

*Canada wrestles with bee-killing crop pesticides
Government recommends mitigation measures, not ban*









*Canadian government scientists have found evidence that neonicotinoid pesticides were linked to mass bee deaths during the spring corn planting in Ontario and Quebec in 2012. (Heinz-Peter Bader/Reuters)*

Canadians beekeepers, farmers and regulators are wrestling with how to protect bees from popular pesticides that were partially banned in Europe this week.

The European Commission announced Monday that it would go ahead with a partial two-year ban on three kinds of neonicotinoid pesticides that have been linked to bee deaths. The pesticides are used to coat most commercial corn seeds and protect them from pests such as seed-eating insects.

Canadian government scientists have found evidence that neonicotinoid pesticides were linked to mass bee deaths during the spring corn planting in Ontario and Quebec in 2012, Health Canada's Pest Management Regulatory Agency confirmed in a report.

Read more about Health Canada's findings HERE:
http://www.cbc.ca/news/canada/windsor/story/2013/04/29/wdr-ontario-farmers-bees-corn-planting.html

*To ban or not to ban?*

That has some people, such as *Dan Davidson, president of the Ontario Beekepers' Association*, calling for the use of the neonicotinoid pesticides to be restricted in Canada also.



> *"I think the best for beekeepers would be a ban," he told CBC's The Current. "We have to call for replacement of these chemicals.
> We won't be able to keep going on if they continue to be used at the rates they're being used now." *


*Listen to the full interviews on 'The Current' *
http://www.cbc.ca/thecurrent/episode/2013/05/03/reversing-the-collapse-of-the-honey-bee-industry/

The environmental advocacy group Sierra Club Canada is similarly calling for a Canada to take the pesticides off the market until they have been proven safe.

However, Kevin Armstrong, a farmer who grows corn, wheat and soybean south of Woodstock, Ont., said neonicotinoid pesticides are essential for protecting corn seeds and seedlings during their crucial first month.

"It is a kind of insurance policy for us," he told The Current. "The vigour of the whole plant is assured for the whole season."

Armstrong said neonicotinoids are largely responsible for a 15 per cent increase in Ontario corn yields over the past 15 years, and so a ban on them could cause a significant loss. A loss of 10 per cent translates into about $100 an acre, he said. If Ontario farmers plant 2.3 million acres of corn as expected, that could amount to a $230-million loss.

"It works out to a significant economic setback for us."

*2012 mass bee-deaths unprecedented*

Mary Mitchell, director-general of the environmental assessment directorate with Health Canada's Pest Management Regulatory Agency, said neonicotinoid pesticides have been registered in Canada for 10 to 15 years and mass bee deaths linked to them had never been reported before last year.

_*"So we do think the weather may have been a factor,"*_ she said, noting that it had been an unusually early, warm dry spring.

She said regulators are working to prevent that happening again, but she did not mention any talk of restrictions on the use of the pesticides.

Instead, she said the government is encouraging farmers to communicate better with beekeepers and to using planting equipment that minimizes the production of dust, which is thought to be a major way bees are exposed to the pesticides.

The government is also working with the agricultural industry on ways to get the pesticide coating to stick better to the seed so it can't come off and harm the bees.

Tracy Baute, who leads the field crop entomology program at the Ontario Ministry of Agriculture and Food, said more studies are underway to find out exactly how bees are exposed to neonicotinoid pesticides.

However, in the meantime, she recommends that farmers:

Let nearby beekeepers know when they are planting so the beekeepers can move hives if necessary.
Consider planting in the early morning or the evening, when bees are less active.
Consider using seeds that aren't treated with pesticides in fields at a lower risk of attack by pests.


The reports of mass bee deaths in Ontario and Quebec in 2012 took place around the time that two scientific studies were published showing that bees can be harmed by even low levels of neonicotinoids.

Many bee species have been declining in North America and Europe, and some have even gone extinct or are believed to be close to extinction. Meanwhile, honeybees have been reported dying or disappearing en masse since 2006. In addition to pesticides, there is evidence that fungi, viruses, or parasites may play a role.


----------



## borderbeeman

*The science behind the eu ban on neonics: Bbc radio programme*

This is an excellent BBC Radio discussion of the Science behind the ban on neonics in Europe.
It is available online in America via this link, which should work in the USA.

*Professor Dave Goulson* (bumblebee expert at Sussex University) is good and *Professor Lynn Dicks of Cambridge University* (entomologist: moth and butterfly expert) is good on the regulatory process.


http://www.bbc.co.uk/programmes/b01s4sz8

*BBC Radio: 'MATERIAL WORLD': Bees and pesticides;*

Duration: 10 minutes

First broadcast: Thursday 02 May 2013"EU states have voted in favour of a proposal to restrict the use of certain pesticides that have been linked to causing serious harm in bees. Neonicotinoid chemicals in pesticides are sprayed onto seeds and spread throughout the plant as it grows. There has been a lot of concern about this systemic approach, with some scientists arguing that it is comparable to using antibiotics 'prophylactically' - every day of your life (in case you get a sore throat). 

Professor Dave Goulson from the University of Sussex and Dr. Lynn Dicks from the University of Cambridge discuss the scientific evidence currently available on these pesticides as well as the limited data available on the population and health of hundreds of other pollinating insect species."​


----------



## borderbeeman

*Re: The science behind the eu ban on neonics: Bbc radio programme*

*Reduction in homing flights in the honey bee Apis mellifera
after a sublethal dose of neonicotinoid insecticides*

Takashi MATSUMOTO
*
Honey Bee Research Unit, NARO Institute of Livestock and Grassland Science, Tsukuba, Japan*

*Abstract*
The negative effects of a commonly applied systemic insecticide, neonicotinoid, on the honey bee Apis mellifera L. are of great concern worldwide, as the use of the chemical is expanding. Recently, special attention has been paid to the sublethal effects of insecticides. An increasing number of studies has identified sublethal effects on the honey bee in the laboratory or in experimental cages, but so far, few studies have examined sublethal effects in the field. To reveal sublethal effects under field conditions, I examined whether the proportion of successful homing flights by foraging honey bees during 30 min after release decreased after bees were topically exposed to insecticides. Honey bees were treated with two types of neonicotinoid insecticide (clothianidin, Dinotefuran) and two types of previously common insecticide (etofenprox [pyrethroid] and fenitrothion [organophosphate]) at five different doses (one-half, one-fourth, one-tenth, one-twentieth, and one-fortieth of their median lethal dose - LD50). 

Then the bees were released 500metres from their hives in the field. The proportions of successful homing flights by bees exposed to neonicotinoids and pyrethroid decreased with doses of one-tenth LD50 (2.18 ng/ head for clothianidin, 7.5 ng/ head for dinotefuran) or more and one-fourth LD50 (32.5 ng/ head for pyrethroid) or more, respectively, whereas bees exposed to organophosphate did not significantly show a response at any sublethal dose though the trend in decline appeared to.

Flight times were not significantly different among treatments at any dose. These results indicate that neonicotinoid and pyrethroid exposure reduced successful homing flights at doses far below the LD50 in the field. Moreover, neonicotinoid caused reductions at relatively lower exposure than pyrethroid.

Key words: nonlethal, pollinator, insecticide, clothianidin, dinotefuran, pyrethroid, etofenprox.


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## VodoBaas1

*CCD in the news again.*

Nothing new, but was a quick read.

http://finance.yahoo.com/news/mass-...Nob21lBHB0A3BtaAR0ZXN0A05hY2VsbGVfT2Zm;_ylv=3


----------



## Barry

*New Science study links bee-killing neonics to mass death of waterlife*

Originally posted by borderbeeman

http://m.guardian.co.uk/environment/...ebrate-die-off



Damian Carrington

The world's most widely used neonicotinoid insecticide is devastating dragonflies, snails and other water-based species, a ground-breaking Dutch study has revealed. http://www.plosone.org/article/info%...l.pone.0062374

On Monday, the insecticide and two others were banned for two years from use on some crops across the European Union, due to the risk posed to bees and other pollinators, on which many food crops rely.

However, much tougher action in the form of a total worldwide ban is needed, according to the scientist who led the new study.

*"We are risking far too much to combat a few insect pests that might threaten agriculture," said Dr Jeroen van der Sluijs at Utrecht University. "This substance should be phased out internationally as soon as possible."*

The pollution was so bad in some places that the ditch water in fields could have been used as an effective pesticide, he said.

Van der Sluijs added that half the 20,000 tonnes of the imidacloprid produced each year is not affected by the EU ban. It is used not to treat crops, but to combat fleas and other pests in cattle, dogs and cats. _* "All this imidacloprid ends up in surface water," *_he said.

The research, published in the peer-reviewed journal PLOS One, found that _*70% less invertebrate species were found in water polluted with the insecticide *_ compared to clean water. There were also far fewer individuals of each species in the polluted water. 

_*"This is the first study to show this happens in the field,"*_ van der Sluijs said. _*"As well as killing mayflies, midges and molluscs, the pollution could have a knock-on effect on birds such as swallows that rely on flying insects for food[/B*_*]", he added.*​*


"Bee-harming pesticides are now leaking into water where they are affecting wildlife," said Friends of the Earth's Paul de Zylva. "This study shows safety levels for chemicals are being routinely breached. Apart from not being properly tested for their risk to bees and other wildlife, pesticides are being used significantly above safe levels and without proper enforcement."​
Julian Little, spokesman for Bayer Cropscience, which manufactures imidacloprid, said:
"There doesn't appear to be anything hugely surprising in this article. It shows the presence of high levels of insecticide in water can have effects on aquatic insects and other invertebrates. Should we have strong stewardship of insecticides to minimise any contamination of water? Yes we should and yes we do."​

The research combined results from wildlife and water pollution surveys at 700 sites across the Netherlands conducted between 1998 and 2009. It found a very strong correlation between high levels of imidacloprid pollution and low numbers of invertebrates. In water exceeding the Dutch national pollution limit, just 17 species were found on average, whereas 50 species were found in cleaner water.

Van der Sluijs said it was highly likely the insecticide was causing the invertebrate die-offs, because imidacloprid was already known to be acutely toxic to these species and is by far the greatest pollutant in the waters.
"Of all the chemicals, it is one of the prime suspects and when you look at the level of exceedence - often 100 times above national limits - it is suspect number one," he said.

The scientists found several cases of extreme pollution, with imidacloprid levels 25,000 times the limit.

"The field-water contained so much insecticide that it could actually be used directly as a lice-control pesticide," van der Sluijs said. "A bee or bumblebee drinking that water would die within a day."​
The extreme cases were all found close to greenhouses, in which imidacloprid is added to the water used to water the plants.

The EU standard for imidacloprid pollution is five times higher than the Dutch limit - 67 nanogram per litre versus 13 ng/l - but even water meeting this standard proved toxic for many species. Water meeting the EU standard has 50% less species that were found in the cleaner water.

Van der Sluijs said the imidacloprid pollution appeared to break existing EU law:

"In my view the present use of imidacloprid is not consistent with what the law says: that the product should not have unnacceptable impacts on non-target organisms."​
He blamed the underlying problem on imidacloprid's extreme potency in killing invertebrates and its long persistence in soil and water. He said there was also a "system error" in the way that pesticides are authorised in the EU, which, for example, assesses only their effect in individual crops, not any cumulative impact.

A recent report by MPs on the UK parliament's green watchdog, the environmental audit committee, concluded that the EU approval process for pesticides was flawed and opaque. "The entire pesticide approval process needs an urgent overhaul," said de Zylva.*


----------



## borderbeeman

USGS has created interactive pesticide-usage maps for the 459 most-used pesticides in the USA - a stupendous piece of work.

hphttp://water.usgs.gov/nawqa/pnsp/usage/maps/compound_listing.php









Imidacloprid use for USA - 2009

Henk Tennekes has commented that there is a correlation between the annual use of Imidacloprid and Clothianidin with the geographical occurrence of CCD; I cannot verify this as I am on vacation and only have intermittent laptop use but it would be a nice project for someone to see if there is a geographical correlation.


----------



## BlueDiamond

*Re: New Science study links bee-killing neonics to mass death of waterlife*

A few days ago Randy Oliver told us why this Dutch study didn't really demonstrate the neonics were responsible for the invertebrate die-offs:
http://community.lsoft.com/scripts/wa-LSOFTDONATIONS.exe?A2=ind1305&L=BEE-L&D=1&O=D&P=122164

Excerpt: "The authors were not out to determine the causes of species decline, but rather only looked to see whether imidacloprid appeared to have any statistical correlation. The authors make this clear. The point being that since imidacloprid is associated with bulb growing, one would expect to find higher concentrations of it in bulb-growing areas (the southwest of the Netherlands). However, in those same areas one would also expect to see high concentrations of some of the other up to 600 different compounds monitored, including insecticides, herbicides, surfactants, fertilizers, etc. The study did not address the contribution by any of these other compounds. I personally have been involved in stream monitoring in California, and have plenty of experience in seeing streams nearly devoid of aquatic life. No pesticides need be involved--simple exposure to fertilizers or road runoff may be enough."


----------



## borderbeeman

*"Unintended Consequences of Field Crop Seed Treatments on Honeybees"*


http://www.soils.wisc.edu/extension/wcmc/2012/ppt/Krupke_2.pdf

Excellent PDF/ Powerpoint from Dr Christian Krupke at Purdue University, Indiana - on the issues centred on neonic-treated field crops and their impact on honey bees through multiple routes of pesticide exposure.


----------



## BigDawg

*Re: Monsanto GMO products contribute to ccd*

http://topinfopost.com/2013/05/28/russia-warns-obama-monsanto

"The shocking minutes relating to President Putin’s meeting this past week with US Secretary of State John Kerry reveal the Russian leaders “extreme outrage” over the Obama regimes continued protection of global seed and plant bio-genetic giants Syngenta and Monsanto *in the face of a growing “bee apocalypse” that the Kremlin warns “will most certainly” lead to world war."*


----------



## BigDawg

*Re: Monsanto GMO products contribute to ccd*

Japan cancels large US wheat order due to recent discovery of GMO contamination of non-GMO wheat fields in Oregon:

http://www.marketwatch.com/story/ja...on-gmo-fear-report-2013-05-30?dist=beforebell


----------



## BigDawg

*Re: Monsanto GMO products contribute to ccd*

Japan has already canceled a large US wheat order today based upon fears of GMO contamination of the US wheat supply and Korea is expected to do the same...

http://www.reuters.com/article/2013/05/30/us-wheat-asia-idUSL3N0EB1JC20130530

"State agriculture department Director Katy Coba said 85 to 90 percent of the Pacific Northwest's soft white wheat crop is exported to Japan, South Korea, Taiwan and other nations, where it's used to make noodles and crackers. Oregon's wheat crop is valued at $300 million to $500 million annually, depending on yield and price.

"Clearly there's a concern about market reaction," Coba said. "Japan and Korea jump out. They do not want genetically-engineered food, they do not want genetically-engineered wheat. They could shut off the market to us."

"A 2005 study estimated that the national wheat industry could lose $94 to $272 million annually if GE wheat were introduced, because many markets oppose or prohibit modified crops, according to the Center for Food Safety."

http://www.oregonlive.com/business/index.ssf/2013/05/genetically_engineered_wheat_f.html#comments


----------



## melliferal

*Re: Monsanto GMO products contribute to ccd*



BigDawg said:


> http://topinfopost.com/2013/05/28/russia-warns-obama-monsanto
> 
> "The shocking minutes relating to President Putin’s meeting this past week with US Secretary of State John Kerry reveal the Russian leaders “extreme outrage” over the Obama regimes continued protection of global seed and plant bio-genetic giants Syngenta and Monsanto *in the face of a growing “bee apocalypse” that the Kremlin warns “will most certainly” lead to world war."*


This entire incident appears to have been made up.


----------



## BlueDiamond

*Re: Monsanto GMO products contribute to ccd*



melliferal said:


> This entire incident appears to have been made up.


And the discovery of GMO wheat in Oregon may have been caused by an act of sabotage: http://finance.yahoo.com/news/monsanto-modified-wheat-isolated-occurrence-160529439.html

"Sabotage is a possibility, said Robb Fraley, Monsanto chief technology officer." "We're considering all options and that's certainly one of the options," Fraley said.


----------



## BigDawg

*AHPA Speaks Out Against GM Bees, Calls For More Research On GMO's*

"Whereas genetically engineered honeybees could have devastating economic implications to the value and the marketability of honey and honeybee products, and enormous economic costs to the bee industry due to the intellectual property rights laws of genetically engineered organisms (beekeepers may not be able to openly breed their own bees because of royalties to a genetic monopoly), therefore be it resolved that the AHPA goes on record as strongly opposing the testing, development, and release of genetically engineered honeybees."

https://c.ymcdn.com/sites/ahpanet.site-ym.com/resource/resmgr/AHPA_Documents/AHPAResolutions2013.pdf


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## Barry

*Are engineered foods evil?—A reply to Scientific American’s David H. Freedman*

*Are engineered foods evil?—A reply to Scientific American’s David H. Freedman*

http://www.geneticliteracyproject.o...ific-americans-david-h-freedman/#.Um6gCSRQ24g


----------



## Barry

*Labels for GMO Foods Are a Bad Idea - Mandatory labels for genetically modified foods*

Labels for GMO Foods Are a Bad Idea - Mandatory labels for genetically modified foods are a bad idea

http://www.scientificamerican.com/article.cfm?id=labels-for-gmo-foods-are-a-bad-idea


----------



## jim lyon

*Re: Labels for GMO Foods Are a Bad Idea - Mandatory labels for genetically modified f*

I think some in the anti-gmo movement have overplayed their hand a bit. Greenpeace's stand on "golden rice" seems to have really fractured that organization. 
http://news.nationalpost.com/2013/1...ified-golden-rice-costing-thousands-of-lives/


----------



## Adrian Quiney WI

*First the bees then the birds?*

This link is from Allen Dick's website. It is about research on the impact of neonics persisting in wetlands and the effect of them on non-targeted water bugs and the birds that feed on them.
http://www.cbc.ca/news/canada/saska...minating-prairie-wetlands-scientist-1.2482082


----------



## Josh Rollins

*Re: First the bees then the birds?*

List of common products anyone can buy from box stores to spray around their house and in the garden. Probably not a full list.

http://www.beyondpesticides.org/pollinators/documents/pesticide_list_final.pdf


----------



## Hilltop

Some interesting new research about plant viruses: http://acsh.org/2014/01/latest-buzz-bee-colony-collapse-disorder-virus-pesticide-problem/


----------



## Barry Digman

Sub-lethal exposure to neonicotinoids impaired honey bees 
winterization before proceeding to colony collapse disorder 



http://www.bulletinofinsectology.org/pdfarticles/vol67-2014-125-130lu.pdf


----------



## Barry Digman

Just to remind folks about this particular thread:




Barry said:


> Use this thread to post Articles, Studies or Links that apply to the topic of CCD.
> This will allow members to have all supporting data in one place.
> 
> This thread is NOT for discussion.
> Post supporting data only. Discuss data in other threads.


----------



## DrStickles

Honeybees are not butterflies, of course. But given the lack of research about the possible effects of herbicides on bee brood, I thought this study of the effect of two herbicides on butterfly larvae might be worthy of note:

*****************************************************************

Journal of Insect Conservation
February 2010, Volume 14, Issue 1, pp 53-63
Date: 29 Apr 2009

*Effects of grass-specific herbicides on butterflies: an experimental investigation to advance conservation efforts*
Cheryl Russell, Cheryl B. Schultz

ABSTRACT
Encroachment by invasive plants is a leading threat to rare butterflies. Restoration plans increasingly recommend herbicides to control invasive plants within butterfly habitats. Few studies address the effects of these herbicides on at-risk butterflies. The effects of two graminicides (fluazifop-p-butyl and sethoxydim) and a surfactant (Preference®) were evaluated on Icaricia icarioides blackmorei and Pieris rapae. The effects on butterfly larvae were assessed by mimicking recommended timing and mixture rates of field applications. Differences in survival to adult eclosure, development time, biomass, sex ratio and adult morphology were assessed. Survival of P. rapae was reduced by 32% with sethoxydim and 21% with fluazifop-p-butyl. Wing size and pupal weights of P. rapae were reduced by herbicide treatments. Icaricia icarioides blackmorei experienced a 21% reduction in development time from the date of treatment to eclosure. These results highlight the importance of careful consideration in the use of herbicides in habitats harboring at-risk butterfly populations.


----------



## squarepeg

on crop losses in the uk for the first season under the 'precautionary principle':

http://risk-monger.blogactiv.eu/201...cautionary-fail-who-is-to-blame/#.VFyuyGeKVps


----------



## BernhardHeuvel

Similiar losses have been reported by the lobbyists in Germany. Interesting - did they really ban neonics in the UK? I just wonder, since this is new to me. 




> Published data indicate that in most cases there is_ no_difference in soybean yield when soybean seed was_treated with neonicotinoids versus not receiving_any insect control treatment. Furthermore,_neonicotinoid seed treatments as currently applied_are only bioactive in soybean_foliage for a period_within the first 3-4_weeks of planting, which does_not overlap with typical_periods of activity for some target pests of concern.[]
> In most cases, these alternatives are comparable in cost to one another and to neonicotinoid seed treatments. The cost of application was considered in this comparison, although because these alternatives can be tank-mixed with other chemicals that are typically applied to soybeans, additional passes over a field would not be necessary. In comparison to the next best alternative pest control measures, neonicotinoid seed treatments likely provide $0 in benefits to growers and at most $6 per acre in benefits (i.e., a 0%-1.7% difference in net operating revenue). Some neonicotinoid seed treatment usage could provide an insurance benefit against sporadic and unpredictable pests, particularly in the southern United States. However, *BEAD did not find information to support the real-world significance of this benefit*, and overall evidence indicates that any such potential benefit is not likely to be large orwidespread in the United States.


from:
*Benefits of Neonicotinoid Seed Treatments to Soybean Production*
UNITED STATES ENVIRONMENTAL PROTECTION AGENCY 
Peer Review Date: October 3, 2014 
http://www2.epa.gov/sites/productio...d_seed_treatments_to_soybean_production_2.pdf


----------



## squarepeg

looks like germany had a good year for rapeseed according to reuter's:

http://in.reuters.com/article/2014/07/30/grain-germany-harvest-idINL6N0Q53XE20140730


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## camero7

New post on Bee-L this morning. worth the read IMO.
http://www.science20.com/jon_entine...s_neonics_may_actually_help_bee_health-149615


----------



## Barry

http://us1.campaign-archive2.com/?u=5fd2b1aa990e63193af2a573d&id=fe62c70514&e=771dfa6e48



> Concern about the survival of the European honeybee has blossomed into a media frenzy during the past several years, with activists declaring, “Beepocalypse”! Beekeepers have seen see some of their honeybee hives disappear in recent years, and concerned observers have blamed the losses on everything from cell phones to genetically modified crops. The most frequently alleged culprit, though, is a class of pesticides known as neonicotinoids. But such alarmism is not supported by the facts. ​



​


----------



## Shumpitron

Naturwissenschaften. 2012 Feb; 99(2): 153–158.
Published online 2012 Jan 13. doi: 10.1007/s00114-011-0881-1
PMCID: PMC3264871

*Pesticide exposure in honey bees results in increased levels of the gut pathogen Nosema*

Jeffery S. Pettis,corresponding author1 Dennis vanEngelsdorp,2 Josephine Johnson,3 and Galen Dively4


http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3264871/


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## BernhardHeuvel

On a German beekeeper and scientist convention the following has been presented. Some studies on neonics and varroa. Findings:


 *Insecticides affect the population dynamics of mites!*
 Insecticides cause stress!
 Significant effects in the fall if varroa + pesticides
 *varroa load is higher in the A and B group* (which was fed with sublethal doses of neonics)
 Feeding of neonicotinoids leads to the reduction of hemocytes.
 Neonics affect wound closure (wound may not heal well).
 For thiacloprid this effect occurs only after high doses. For Clothianidin the effects occur after low doses.

Original document (in German)
http://imker.cwsurf.de/index.php/downloads/category/3-dokumente?download=5:tagung


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## camero7

http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0118748

Conclusions To our knowledge, this study is the first to examine the chronic sublethal effects on whole honey bee colonies subjected to worse-case scenarios as well as normal dietary exposure (5μg/kg) to imidacloprid. We used spiked diet patties placed within colonies to deliver continuous direct exposure over multiple brood cycles to imidacloprid residues that were generally higher than levels found in bee-collected pollen and nectar under field conditions. Our results provide evidence that imidacloprid exposure doses up to 100 μg/kg had no significant effects on foraging activity or colony performance during and shortly after 12 weeks of exposure. However, several colony performance endpoints showed dose-response patterns, particularly higher _Varroa_ infestations with increased dose, though not all patterns were statistically significant. The major finding was the higher rates of queen replacement and resulting broodless periods during the late summer in colonies exposed to 20 and 100 μg/kg of imidacloprid, which led to weaker colonies going into the winter. These exposure regimes sublethally affected colony health and significantly reduced overwintering success. However, the question remains as to whether doses of 100 or even 20 μg/kg exposed for 12 continuous weeks realistically represent imidacloprid residues in bee-collected food under agriculture settings. In certain field situations, residues of imidacloprid can reach or exceed 100 μg/kg in pollen of treated crops during several weeks of flowering [35] or in guttation droplets exuded from treated corn seedlings [45,46]. However, it is uncommon for honey bees to be exposed to these doses for extended periods. Furthermore, bees generally forage on different water and floral sources simultaneously and not all sources will contain residues; thus their foraging behavior tends to reduce the concentration of imidacloprid in food stored in the colony. The within-hive fate experiment demonstrated that imidacloprid residues of 100 μg/kg in diet patties or 20 μg/kg in sucrose syrup became diluted or non-detectable due to the processing of beebread and honey and the rapid metabolism of the chemical by bees. Given the weight of evidence presented here, we conclude that chronic exposure to imidacloprid at the higher range of field doses (20 to 100 μg/kg) in the pollen of certain treated crops could contribute to reduced overwintering success but the most likely encountered field doses of 5 μg/kg, especially relevant for seed-treated crops, have negligible effects on honey bee colony health. Currently there is wide agreement that sublethal exposure to imidacloprid can cause adverse effects on honey bees in laboratory studies [77] but no evidence that this widely used insecticide is the major stressor causing colony declines. Our findings agree with a causal analysis by Staveley et al. [37] that judged neonicotinoid pesticides to be an unlikely sole cause of colony declines. Finally, this study makes evident the importance of conducting risk assessment studies on honey bee colonies over longer periods to reveal the chronic sublethal effects on queen health and bee behaviors that can ultimately impair colony performance


----------



## suburbanrancher

I don't think I've seen this one posted here: 
http://phys.org/news/2015-02-neonicotinoid-insecticides-impair-bee-brains.html


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## camero7

New study on neonics.

http://msucares.com/news/print/agnews/an15/20150617_1.html


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## camero7

A well-balanced paper on neonics and honey bees.
http://www.nature.com/nature/journal/v521/n7552_supp/full/521S52a.html


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## Barry

Effects of Sublethal Doses of Imidacloprid on Young Adult Honeybee Behaviour

http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0140814


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## camero7

With all due respect, this is another cage study of forced poisoning. I really don't believe it correlates to the real world IMO.


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## Barry

It doesn't!


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## BernhardHeuvel

*Honeybees Produce Millimolar Concentrations of Non-Neuronal Acetylcholine for Breeding: Possible Adverse Effects of Neonicotinoids*
Ignaz Wessler , Hedwig-Annabel Gärtner, Rosmarie Michel-Schmidt, Christoph Brochhausen, Luise Schmitz, Laura Anspach, Bernd Grünewald, Charles James Kirkpatrick
Published: June 10, 2016
http://dx.doi.org/10.1371/journal.pone.0156886
http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0156886


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## camero7

Poor study IMO. Nothing here.


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## BernhardHeuvel

Poor answer. No reasons pointed out for me to understand. Don't you want people to understand why you think, this is a poor study?


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## camero7

test tube study, nothing that relates to the real world. Not realistic dosages of neonics.


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## BernhardHeuvel

Ok, fine. I just wait for the absolute definition of realistic dosages found outside in the field. 

At least I find it interesting that ACh is in use outside the neural system. And the importance for the development of the larvae.


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## camero7

There is no doubt that neonics are poisonous to bees. It's just my opinion that they are less deadly than anything that can replace them right now. I don't delude myself that farmers are going to give up pesticides and having seen the devastation to hive from the organophosphates, I'm pretty happy that I have never seen that from neonics. The only hives I've lost to pesticides is from a idiot spraying apples during the day while in bloom. Even then the hives survived for a while. My father lost about 50 hives in about 20 minutes when I was a kid. Dead bees everywhere, you could scoop them up with a shovel.


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## BernhardHeuvel

*Performance of honeybee colonies located in neonicotinoid-treated and untreated cornfields in Quebec*

M. Alburaki1,2,†,*, B. Cheaib1, L. Quesnel1, P.-L. Mercier1,2, M. Chagnon3 andN. Derome1,4
Version of Record online: 2 JUN 2016

DOI: 10.1111/jen.12336

"Overall, our results show that forager bees collected 20% of corn pollen containing variable concentrations of neonicotinoids. Colonies located in treated cornfields expressed higher varroa loads and long-term mortality than those in untreated cornfields."
http://onlinelibrary.wiley.com/doi/...nticated=false&deniedAccessCustomisedMessage=


----------



## johno

Bees that are collecting feed corn pollen are starving.
Johno


----------



## BernhardHeuvel

Johno,

that's true. But bees forage for corn pollen here on a regular basis. I already posted pictures and videos on beesource some time ago. 


*Neonicotinoid-contaminated pollinator strips adjacent to cropland reduce honey bee nutritional status*
Christina L. Mogren & Jonathan G. Lundgren

Scientific Reports 6, Article number: 29608 (2016) doi:10.1038/srep29608
Published online: 14 July 2016

citation: 
"Increasing concentrations of clothianidin in bee bread were correlated with decreased glycogen, lipid, and protein in workers. This study shows that small, isolated areas set aside for conservation do not provide spatial or temporal relief from neonicotinoid exposures in agricultural regions where their use is largely prophylactic."
http://www.nature.com/articles/srep29608


----------



## Michael Palmer

BernhardHeuvel said:


> " Colonies located in treated cornfields expressed higher varroa loads and long-term mortality than those in untreated cornfields."


I can't dispute the finding, but my apiaries are surrounded by clothianadin corn and I'm not seeing what they say they are finding. The states of New York and Vermont have been testing my colonies, looking for varroa and nosema. Varroa loads in July have been 0-2. 

My cell building apiary is a good example. Surrounded by corn. 35 very strong colonies. 64 very strong brood factory nucs. Made enough queen cells to produce about 1200 mated queens. Brood factories gave me enough brood to set up 56 cell builders, and about 200 4 frame nucs for wintering. 

Now, I've used this apiary for years as my cell building yard and as a brood source for most of my nucs. Don't you think, if treated corn was a real problem for my bees, I would know? And please, don't play the sublethal card.


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## BernhardHeuvel

Do you feed your cell builders and supporting colonies? I mean, during the season.


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## BernhardHeuvel

A good beekeeper can make up for a lot of problems. And you are a good beekeeper, Michael Palmer. 

I visited a friend some days ago, he's a master beekeeper (and professional beekeeper, 3rd generation). What we found is this: 










Upper part of the body is more brown than shown in this archive picture. Abdomen is white.

What would you say this is? We'd say it is chilling of the brood. But how come? Formerly thriving hives, boiling with bees. Suddenly spotty broodnests, and the pupae look like this. We opened almost every single broodcell in those hives. We did not found any mites, absolutely zero.

What is it? What's wrong with those hives?










Even queens.










You see the difference between the cappings of the outer and inner brood?


----------



## jim lyon

I don't know, are you suggesting it could only be neonics? I have seen some fungicide issues that resembled this. I do know it's something any good beekeeper would want an answer for.


----------



## Michael Palmer

BernhardHeuvel said:


> Do you feed your cell builders and supporting colonies? I mean, during the season.


No, only some feed to the cell builders when I graft, but not otherwise. In fact, this year the sumac flow was so intense that my breeder colonies filled the grafting comb with nectar as soon as I gave it to them. Made it difficult to find graft-able larvae.


----------



## BernhardHeuvel

jim lyon said:


> I don't know, are you suggesting it could only be neonics?


No, of course not. In this case shown above that are pictures from the poisoning incident some time ago. Bees were tested in the state labs and they found clothianidin as the causing pesticide. Bayer paid compensation to the beekeepers and this company usually doesn't do this quickly. It simply was the fault of that pesticide. 

So if you find this like you see in the pictures, this should be some sort of poisoning. We have seen this before with the pesticide Insegar, so yes, it is not only neonics. But too often neonics were involved. We don't only talk, we get it laboratory tested and thus proofed. Problem is, that doesn't help much with the overall situation. Only thing we can do, is to prevent the foraging on neonic crops as good as we can. A friend took his bees into the forest, only to find out, that the christmas trees grown there were sprayed with Karate Forst, another neonic. He lost 50 hives. 

It is heart breaking when you open your hives and find this sh*t shown above. You do your best to keep the bees in a perfect shape. And you end up with trouble over and over again. Trouble you can't really escape.


----------



## Michael Palmer

Bernhard, I'm not trying to disprove what you say. I'm only wondering out loud why I don't see such clothianadin poisoning in my bees. Is it that the poisoned colonies had nothing to forage on but corn?


----------



## johno

I have never seen neonics sprayed in my area, only seed coatings are used. I would suspect that neonics could be sprayed onto cotton south of my location and that would probably cause bee losses. However I am in a dearth at the moment until maybe September and I check the corn and soybean fields and have never seen bees feeding in either. So Bernhard you should ask yourself what are your farmers and beekeepers doing in Germany that is different to what most successful beekeepers are doing in the USA. I have seen references made about the Europeans use a far higher dosage of neonics than the American farmers do.
Johno


----------



## BernhardHeuvel

The difference is landscape. We don't have the open country and wildnerness you have. We are crowded here and little to no nature is left. Also rain and ground water is different here.

At the time corn is flowering there is not much else out there for the bees. No nectar flow at all and only little pollen from other plants. At times corn pollen makes up 70-90 % of total pollen consumption. A steady flow of light yellow pollen. 

Being so crowded here, we have the following situation. 

I) Germany has 357,167.94 km² - that are 357,167,940,000 m². 

II) *100 tons* of one neonic alone: Imidacloprid are applicated here each year - that equals 100.000.000.000 mg per year. 220462.2621848776 pounds.

III) Divide this all over Germany and you get 0.279980336421 mg per square meter.

IV) 0.279980336421 mg are 279,980.336421 ng. Per square meter!

V) The chronic-lethal dosis (LD50 chronic) for bees is 0.01-1 ng per honeybee. (Suchail et al. 2001)

And that is only one neonic.

That is simply an "overkill". Probably the stuff dilutes into your open space country where you live. Here it builds up and concentrates more and more.

How much is totally applied in your country, let's say Imidacloprid and what is the overall acreage?

There must be differences because of the different observations. Sure. But we know exactly what is causing our problems. We did our homework.


----------



## crofter

Here is a link to newspaper article on Canadian ovewinter losses. I was surprised to see starvation listed as #1 cause. Not necessarily very scientific.

http://kitchener.ctvnews.ca/16-8-of-honeybee-colonies-lost-over-winter-survey-finds-1.2998795


----------



## johno

This is another story that does not seem to support the theory that neonics are the cause of all the bee problems. A great deal of Canadians put their bees into neonic raised canola without many bee losses, but then again canola pollen is high in protein and is supposed to be good for bee nutrition. Neonics might just speed up the demise of under nourished bees, just a thought.
Johno


----------



## johno

Bernhardt, From a thread you posted on another forum in which you mention the amount of your harvest makes it hard to believe you have starving bees feeding on corn pollen. http://www.beesource.com/forums/sho...arm-Control-Made-Very-Easy-–-Apparently/page6 
I have one harvest a year with nectar flow of about 6 weeks in the spring and still my bees do not feed on corn pollen.
I know our bees will go to sweetcorn but there are no neonics used here on sweetcorn.
Johno


----------



## BernhardHeuvel

Michael Palmer, I just saw your presentation on wintering in the North. Well, what shall I say. You say, we don't have a clue what wintering in the North means. True. But I was not only amazed about the snow pile pictures and morning doves, I also was in awe about the blooming meadows and fields and how long your nectar season is compared to ours.

You sure not know what it means to keep bees in the green deserts as we don't know not much about wintering in the far North.

PS: Your comment about insulation: Don't forget that somewhere else there is winter but no snow. Just frost and freezing, but no snow. With all the snow piled up on the hives, you have tons of insulation. Low temperatures and no snow = no insulation is a different matter.


----------



## squarepeg

http://www.tandfonline.com/doi/abs/10.1080/00218839.2016.1162978


----------



## BernhardHeuvel

*Combined neonicotinoid pesticide and parasite stress alter honeybee queens’ physiology and survival*
Claudia Dussaubat, Alban Maisonnasse, Didier Crauser, Sylvie Tchamitchian, Marc Bonnet, Marianne Cousin, André Kretzschmar, Jean-Luc Brunet*& Yves Le Conte
Scientific Reports 6, Article*number:*31430 (2016)
doi:10.1038/srep31430
Published online: 31 August 2016

http://www.nature.com/articles/srep31430

Abstract
Honeybee colony survival strongly relies on the queen to overcome worker losses exposed to combined stressors like pesticides and parasites. Queen’s capacity to withstand these stressors is however very little known. The effects of the common neonicotinoid pesticide imidacloprid in a chronic and sublethal exposure together with the wide distributed parasite Nosema ceranae have therefore been investigated on queen’s physiology and survivorship in laboratory and field conditions. Early physiological changes were observed on queens, particularly the increase of enzyme activities (catalase [CAT] and glutathione-S-transferase [GST] in the heads) related to protective responses to xenobiotics and oxidative stress against pesticide and parasite alone or combined. Stressors also alter the activity of two other enzymes (carboxylesterase alpha [CaE α] and carboxylesterase para [CaE p] in the midguts) involved in metabolic and detoxification functions. Furthermore, single and combined effects of pesticide and parasite decrease survivorship of queens introduced into mating hives for three months. Because colony demographic regulation relies on queen’s fertility, the compromise of its physiology and life can seriously menace colony survival under pressure of combined stressors.


----------



## BernhardHeuvel

*Neonicotinoid pesticides severely affect honey bee queens*
Geoffrey R. Williams, Aline Troxler, Gina Retschnig, Kaspar Roth, Orlando Yañez, Dave Shutler, Peter Neumann*& Laurent Gauthier
Scientific Reports 5, Article*number:*14621 (2015)
doi:10.1038/srep14621
Published online: 13 October 2015

http://www.nature.com/articles/srep14621


----------



## BernhardHeuvel

A new study has found neonicotinoids, the world’s most commonly used insecticide, cause queen honeybees to lay as much as two-thirds fewer eggs, jeopardizing the health and stability of entire bee colonies. 

http://e360.yale.edu/digest/insecticide_neonicotinoids_queen_bee_eggs/4801/


----------



## BernhardHeuvel

"Our results strongly suggest that neonicotinoids can negatively affect honeybee drone sperm quality."

*Sperm parameters of honeybee drones exposed to imidacloprid*

Ciereszko, A., Wilde, J., Dietrich, G.J. et al. Apidologie (2016). doi:10.1007/s13592-016-0466-2
http://link.springer.com/article/10.1007/s13592-016-0466-2

Abstract
The objective of this study was to evaluate the effects of chronical exposure of honeybee drones to environmental (5 ppb) and non-environmental concentration (200 ppb) of imidacloprid (IMD) on sperm concentration, motility, viability, and mitochondrial membrane potential measured in semen obtained from 180 drones originating from 18 colonies. The results demonstrate that IMD exposure did not affect sperm concentration; however, there were significant differences in concentration within colonies. IMD exposure was associated with reductions in sperm motility, which also varied within colonies. Statistically significant interactions between IMD exposure and colony were found for active mitochondria and sperm viability. Our results strongly suggest that neonicotinoids can negatively affect honeybee drone sperm quality. It is important to emphasize that IMD actions can be strongly modulated according to the colony.


----------



## BernhardHeuvel

*Effects of sublethal concentration of imidacloprid on the insulin/insulin-like signaling pathway in the honey bee, Apis mellifera*

http://db.koreascholar.com/article?code=317933

Abstract
Colony collapse disorder (CCD), a phenomenon of honeybees disappearance, has been reported since 2006. Chronic exposure to neonicotinoid insecticides, particularly imidacloprid, has been suggested to impair forager’s ability for foraging and be a main cause of CCD. Recently, it has been reported that imidacloprid induces insulin resistance in animal cell line by blocking glucose uptake. Similarly to human insulin, insulin-like peptide (ILP) of insects is involved in maintaining blood glucose contents in hemolymph by regulating the concentration of trehalose and glycogen. Therefore, we have hypothesized that sublethal concentration of neonicotinoid may affect the metabolic pathway of honey bees as well. We investigated the transcription levels of the genes involved in the insulin/insulin-like signaling (IIS) pathway, such as AmILP and AmInR, following an acute or a chronic dietary exposure of sublethal concentrations of imidacloprid to foragers. In both experiments, honeybees showed increased expression levels of ILP and InR in a dose-dependent manner. Our results suggest that sublethal dose of imidacloprid likely upregulates IIS pathway, thereby rendering honey bees to become resistant to insulin.


----------



## BernhardHeuvel

Another scientific review on neonics:
https://www.degruyter.com/downloadpdf/j/jas.2016.60.issue-2/jas-2016-0024/jas-2016-0024.xml


----------



## BernhardHeuvel

*Sublethal doses of neonicotinoid imidacloprid can interact with honey bee chemosensory protein 1 (CSP1) and inhibit its function*
Hongliang Li, Jing Tan, Xinmi Song, Fan Wu, Mingzhu Tang, Qiyun Hua, Huoqing Zheng, Fuliang Hu
Biochemical and Biophysical Research Communications, Available online 14 March 2017, ISSN 0006-291X, http://dx.doi.org/10.1016/j.bbrc.2017.03.051.
(http://www.sciencedirect.com/science/article/pii/S0006291X17305090)

Abstract: Abstract
As a frequently used neonicotinoid insecticide, imidacloprid can impair the chemoreceptive behavior of honey bees even at sublethal doses, while the physiochemical mechanism has not been further revealed. Here, multiple fluorescence spectra, thermodynamic method, and molecular docking were used to study the interaction and the functional inhibition of imidacloprid to the recombinant CSP1 protein in Asian honey bee, Apis cerana. The results showed that the fluorescence intensity (λem = 332 nm) of CSP1 could be significantly quenched by imidacloprid in a dynamic mode. During the quenching process, ΔH > 0, ΔS > 0, indicating that the acting forces of imidacloprid with CSP1 are mainly hydrophobic interactions. Synchronous fluorescence showed that the fluorescence of CSP1 was mainly derived from tryptophan, and the hydrophobicity of tryptophan decreased with the increase of imidacloprid concentration. Molecular docking predicted the optimal pose and the amino acid composition of the binding process. Circular dichroism (CD) spectra showed that imidacloprid reduced the α-helix of CSP1 and caused the extension of the CSP1 peptide chain. In addition, the binding of CSP1 to floral scent β-ionone was inhibited by nearly 50% of the apparent association constant (KA) in the presence of 0.28–2.53 ng/bee of imidacloprid, and the inhibition rate of nearly 95% at 3.75 ng/bee of imidacloprid at sublethal dose level. This study initially revealed the molecular physiochemical mechanism that sublethal doses of neonicotinoid still interact and inhibit the physiological function of the honey bees' chemoreceptive system.
Keywords: Apis cerana; Chemosensory protein; Neonicotinoid imidacloprid; Binding interaction; Functional inhibition


Consequences of the impairment?


----------



## Bob Nelson

*Documentation of low level neonicotinoids in treated domestic water*

Media report:

https://www.washingtonpost.com/news...des-in-drinking-water/?utm_term=.b8cbd77212d7

Abstract:

http://pubs.acs.org/doi/abs/10.1021/acs.estlett.7b00081

The paper:

http://pubs.acs.org/doi/pdf/10.1021/acs.estlett.7b00081


----------



## GreenbirdAcres

*Re: Effects of neonicotinoid insecticides on mammalian nicotinic acetylcholine recept*

404'd, Barry. Bad link :scratch:


----------



## BernhardHeuvel

*Re: Effects of neonicotinoid insecticides on mammalian nicotinic acetylcholine recept*

Sci Rep. 2017 Apr 26;7(1):1201. doi: 10.1038/s41598-017-01361-8.

*A common neonicotinoid pesticide, thiamethoxam, impairs honey bee flight ability.*


Tosi S1,2,3, Burgio G4, Nieh JC5.
Abstract



Pesticides can pose environmental risks, and a common neonicotinoid pesticide, thiamethoxam, decreases homing success in honey bees. Neonicotinoids can alter bee navigation, but we present the first evidence that neonicotinoid exposure alone can impair the physical ability of bees to fly. We tested the effects of acute or chronic exposure to thiamethoxam on the flight ability of foragers in flight mills. Within 1 h of consuming a single sublethal dose (1.34 ng/bee), foragers showed excitation and significantly increased flight duration (+78%) and distance (+72%). Chronic exposure significantly decreased flight duration (-54%), distance (-56%), and average velocity (-7%) after either one or two days of continuous exposure that resulted in bees ingesting field-relevant thiamethoxam doses of 1.96-2.90 ng/bee/day. These results provide the first demonstration that acute or chronic exposure to a neonicotinoid alone can significantly alter bee flight. Such exposure may impair foraging and homing, which are vital to normal colony function and ecosystem services.

https://www.ncbi.nlm.nih.gov/pubmed/28446783
​


----------



## Oldtimer

*Re: Effects of neonicotinoid insecticides on mammalian nicotinic acetylcholine recept*



BernhardHeuvel said:


> These results provide the first demonstration that acute or chronic exposure to a neonicotinoid alone can significantly alter bee flight. Such exposure may impair foraging and homing, which are vital to normal colony function and ecosystem services.


I'll bet acute or chronic exposure to ANY insecticide can alter bee flight. Or may impair foraging and homing.


----------



## camero7

*Re: Effects of neonicotinoid insecticides on mammalian nicotinic acetylcholine recept*

You'd think, after all these years of neonic hysteria, people would figure out that varroa, nosema and virus are the real problem.


----------



## BernhardHeuvel

*Chronic exposure to neonicotinoids reduces honey bee health near corn crops*

*Chronic exposure to neonicotinoids reduces honey bee health near corn crops *

N. Tsvetkov1, O. Samson-Robert2, K. Sood1, H. S. Patel1, D. A. Malena1, P. H. Gajiwala1, P. Maciukiewicz1, V. Fournier2, A. Zayed1,* + See all authors and affiliations Science 30 Jun 2017: Vol. 356, Issue 6345, pp. 1395-1397 DOI: 10.1126/science.aam7470 

http://science.sciencemag.org/content/356/6345/1395​
Abstract

Experiments linking neonicotinoids and declining bee health have been criticized for not simulating realistic exposure. Here we quantified the duration and magnitude of neonicotinoid exposure in Canada’s corn-growing regions and used these data to design realistic experiments to investigate the effect of such insecticides on honey bees. Colonies near corn were naturally exposed to neonicotinoids for up to 4 months—the majority of the honey bee’s active season. Realistic experiments showed that neonicotinoids increased worker mortality and were associated with declines in social immunity and increased queenlessness over time. We also discovered that the acute toxicity of neonicotinoids to honey bees doubles in the presence of a commonly encountered fungicide. Our work demonstrates that field-realistic exposure to neonicotinoids can reduce honey bee health in corn-growing regions.


----------



## BernhardHeuvel

*Re: Effects of neonicotinoid insecticides on mammalian nicotinic acetylcholine recept*



camero7 said:


> You'd think, after all these years of neonic hysteria, people would figure out that varroa, nosema and virus are the real problem.


All those scientists are wrong, plain wrong. Varroa, varroa, varroa is the problem, right. :scratch:


----------



## johno

*Re: Effects of neonicotinoid insecticides on mammalian nicotinic acetylcholine recept*

Ja Bernhard, especially the scientists in the link you provided as the link states page not found. Hard to check the facts. No!
Johno


----------



## Oldtimer

*Re: Effects of neonicotinoid insecticides on mammalian nicotinic acetylcholine recept*

Try http://science.sciencemag.org/content/356/6345/1393


----------



## johno

*Re: Effects of neonicotinoid insecticides on mammalian nicotinic acetylcholine recept*

OT that is a different study about oilseed rape in 3 different countries, bees did badly in the UK and Hungary but somehow did well in Germany. The one mentioned by Bernhard was carried out in Canada in cornfield areas. This might be in that province where losses were very high and I remember one of those beekeepers mentioning his bees bringing in pollen from the corn. It was suggested if that was the case then in all probability his bees were probably starving. 2017 has been the poorest spring I have seen in the 7 years I have kept bees with just about 4 weeks of flow and then only about 5 acres of crops I have planted for them in my home area. Took in very little honey from 4 outyards and the bees have very little stores so will have to feed heavily to get them stores for winter. However I have hundreds of acres of corn all around my home area and have yet to see any of my bees in the cornfields.
Johno


----------



## Oldtimer

*Re: Effects of neonicotinoid insecticides on mammalian nicotinic acetylcholine recept*

Oops 

I've left the link just so there is context to your interesting comments.


----------



## 1102009

*Re: Effects of neonicotinoid insecticides on mammalian nicotinic acetylcholine recept*



johno said:


> bees did badly in the UK and Hungary but somehow did well in Germany.


@bernhard
I´m not able to read all the scientific work.
Do you know why this differences are? Is it the mixture of neocotinoides with other chemicals or how much is used or the time of day it is used or the technic?
Thanks, Sibylle


----------



## johno

*Re: Effects of neonicotinoid insecticides on mammalian nicotinic acetylcholine recept*

Some would say "location location location " all beekeeping is local. The opinions are that poorly nourished bees do not fare well against stresses produced by insecticides and mite vectored viruses and other diseases. Diversity in bee forage appears to be key to a great deal of bee problems.
Johno


----------



## 1102009

*Re: Effects of neonicotinoid insecticides on mammalian nicotinic acetylcholine recept*



johno said:


> Some would say "location location location " all beekeeping is local. The opinions are that poorly nourished bees do not fare well against stresses produced by insecticides and mite vectored viruses and other diseases. Diversity in bee forage appears to be key to a great deal of bee problems.
> Johno


Could be but locale changes in the same country too.


----------



## BernhardHeuvel

*Re: Effects of neonicotinoid insecticides on mammalian nicotinic acetylcholine recept*

Rather local politics. Certain pesticide company headquarters are located in Germany. It is said, they have some influence on certain scientific institutions (and politicians, journalists,...)
I heared. :shhhh:


----------



## gww

*Re: Effects of neonicotinoid insecticides on mammalian nicotinic acetylcholine recept*

bernhard
That is the position that the pbs new broadcast took when they reported on it last night. They said bayer paid for the study in germany.
Cheers
gww


----------



## 1102009

*Re: Effects of neonicotinoid insecticides on mammalian nicotinic acetylcholine recept*



BernhardHeuvel said:


> Rather local politics. Certain pesticide company headquarters are located in Germany. It is said, they have some influence on certain scientific institutions (and politicians, journalists,...)
> I heared. :shhhh:




I heared Randolf Menzel was involved. He is known but I don´t know which side he is on. He seems to go against the neonics though.
Since the deal bayer-monsanto some problems seem to develop for bayer because of california law. ( since they are under US law now). Who knows maybe it does´t pay in future for them.


----------



## johno

*Re: Effects of neonicotinoid insecticides on mammalian nicotinic acetylcholine recept*

I believe Bayer and Syngenta paid for most of the whole study and that the reason that the German bees were not effected was because they were healthier bees!
Johno


----------



## 1102009

*Re: Effects of neonicotinoid insecticides on mammalian nicotinic acetylcholine recept*



johno said:


> I believe Bayer and Syngenta paid for most of the whole study and that the reason that the German bees were not effected was because they were healthier bees!
> Johno


Nice to know. 
Maybe the observed ones were TF survivors.:scratch:
I have yet to see treated hives here which survive one season being TF. Health could really be an illusion.


----------



## johno

*Re: Effects of neonicotinoid insecticides on mammalian nicotinic acetylcholine recept*

From what I can gather from B-L archives the paper involved is behind a pay wall and not many have actually read the paper, what has been read are the articles published by organizations that have a biased opinion based on their own agenda. There are some who feel that the conclusions reached are within colony variables and in fact state that the UK side was of not much value as most of the controls as well as the neonic fed bees did not survive the winter. So in fact the results like beauty is in the eye of the beholder.
Johno


----------



## johno

*Re: Effects of neonicotinoid insecticides on mammalian nicotinic acetylcholine recept*

SiWolke, yes perhaps the colonies in the UK were TF survivors. And yes I have yet to see a TF hive survive a winter in my area.
See what I mean about the eye of the beholder.
Johno


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## BernhardHeuvel

*Gene expression changes in honey bees induced by sublethal imidacloprid exposure duri*

*Gene expression changes in honey bees induced by sublethal imidacloprid exposure during the larval stage *

Ming-Cheng Wua, Yu-Wen Changa, Kuang-Hui Lub, En-Cheng Yanga

Insect Biochemistry and Molecular Biology
https://doi.org/10.1016/j.ibmb.2017.06.016

Abstract

Honey bee larvae exposed to sublethal doses of imidacloprid show behavioural abnormalities as adult insects. Previous studies have demonstrated that this phenomenon originates from abnormal neural development in response to imidacloprid exposure. Here, we further investigated the global gene expression changes in the heads of newly emerged adults and observed that 578 genes showed more than 2-fold changes in gene expression after imidacloprid exposure. This information might aid in understanding the effects of pesticides on the health of pollinators. For example, the genes encoding major royal jelly proteins (MRJPs), a group of multifunctional proteins with significant roles in the sustainable development of bee colonies, were strongly downregulated. These downregulation patterns were further confirmed through analyses using quantitative reverse transcription-polymerase chain reaction on the heads of 6-day-old nurse bees. To our knowledge, this study is the first to demonstrate that sublethal doses of imidacloprid affect mrjp expression and likely weaken bee colonies.

http://www.sciencedirect.com/science/article/pii/S0965174817301005


----------



## BernhardHeuvel

*Re: Gene expression changes in honey bees induced by sublethal imidacloprid exposure*

Oh...just another one...

Bulletin of Environmental Contamination and Toxicology, pp 1–6

*Impact of Thiamethoxam on Honey Bee Queen (Apis mellifera carnica) Reproductive Morphology and Physiology*

Ivana Tlak Gajger, Martina Sakač, Aleš Gregorc

Abstract

High honey bee losses around the world have been linked in part by the regular use of neonicotinoids in agriculture. In light of the current situation, the aim of this study was to investigate the effects of thiamethoxam on the development of the reproductive system and physiology in the honey bee queen. Two experimental groups of honey bee queen larvae were treated with thiamethoxam during artificial rearing, applied via artificial feed in two cycles. In the first rearing cycle, honey bee larvae received a single treatment dose (4.28 ng thiamethoxam/queen larva on the 4th day after larvae grafting in artificial queen cells), while the second honey bee queen rearing cycle received a double treatment dose (total of 8.56 ng thiamethoxam/queen larva on the 4th and 5th day after larvae grafting in artificial queen cells). After emerging, queens were anesthetized and weighed, and after mating with drones were anesthetized, weighed, and sectioned. Ovary mass and number of stored sperm were determined. Body weight differed between untreated and treated honey bee queens. The results also show a decrease in the number of sperm within honey bee queen spermathecae that received the double thiamethoxam dose.

https://link.springer.com/article/10.1007/s00128-017-2144-0​


----------



## Oldtimer

*Re: Gene expression changes in honey bees induced by sublethal imidacloprid exposure*

A developing queen bee larva is fed poison, and later after it has mated it looks like the poison had a negative effect. Why would that be a surprise?


----------



## BernhardHeuvel

*Re: Gene expression changes in honey bees induced by sublethal imidacloprid exposure*

Not to me. But for some it is still a surprise that a highly toxic and systemic insecticide group is poisonous to bees in field application.

As Bayer said: chemical plus nature.


----------



## 1102009

*Re: Gene expression changes in honey bees induced by sublethal imidacloprid exposure*

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4284386/



> Furthermore, their mode of action enables new strategies for pest control that profit from the existing synergies between these substances and either other chemicals or microorganisms. As a result, there are a wide range of uses available, including seed coating and root bathing, as invertebrate pest control in agriculture, horticulture, orchards, forestry, veterinary applications, and fish farming. However, these same properties have led to problems. Specifically, their widespread (Main et al. 2014) and prophylactic use, their systemic properties in plants, their broad spectrum of toxicity in invertebrates, and the persistence and environmental fate of parent compounds and metabolites renders them potentially harmful to a broad range of non-target organisms.


----------



## 1102009

*Re: Gene expression changes in honey bees induced by sublethal imidacloprid exposure*

http://www.nature.com/articles/s41559-017-0260-1


----------



## BernhardHeuvel

*Planting of neonicotinoid-coated corn raises honey bee mortality and sets back colony*

*Planting of neonicotinoid-coated corn raises honey bee mortality and sets back colony development.*

Abstract 
Worldwide occurrences of honey bee colony losses have raised concerns about bee health and the sustainability of pollination-dependent crops. While multiple causal factors have been identified, seed coating with insecticides of the neonicotinoid family has been the focus of much discussion and research. Nonetheless, few studies have investigated the impacts of these insecticides under field conditions or in commercial beekeeping operations. Given that corn-seed coating constitutes the largest single use of neonicotinoid, our study compared honey bee mortality from commercial apiaries located in two different agricultural settings, i.e. corn-dominated areas and corn-free environments, during the corn planting season. Data was collected in 2012 and 2013 from 26 bee yards. Dead honey bees from five hives in each apiary were counted and collected, and samples were analyzed using a multi-residue LC-MS/MS method. Long-term effects on colony development were simulated based on a honey bee population dynamic model. Mortality survey showed that colonies located in a corn-dominated area had daily mortality counts 3.51 times those of colonies from corn crop-free sites. Chemical analyses revealed that honey bees were exposed to various agricultural pesticides during the corn planting season, but were primarily subjected to neonicotinoid compounds (54% of analysed samples contained clothianidin, and 31% contained both clothianidin and thiamethoxam). Performance development simulations performed on hive populations’ show that increased mortality during the corn planting season sets back colony development and bears contributions to collapse risk but, most of all, reduces the effectiveness and value of colonies for pollination services. Our results also have implications for the numerous large-scale and worldwide-cultivated crops that currently rely on pre-emptive use of neonicotinoid seed treatments.

Samson-Robert O, Labrie G, Chagnon M, Fournier V. (2017)
PeerJ5:e3670 https://doi.org/10.7717/peerj.3670


----------



## Baja

Thanks for the great selection of scientific studies. Here is a collection of about 100 more studies confirming the negative impact of neonicotinoid pesticides on bees. http://strathconabeekeepers.blogspot.ca/p/the-beekeepers-library.html#pesticidesandbees


----------



## BernhardHeuvel

*Sublethal effects of imidacloprid on targeting muscle and ribosomal protein related genes in the honey bee Apis mellifera L.*

Yan-Yan Wu, Qi-Hua Luo, Chun-Sheng Hou, Qiang Wang, Ping-Li Dai, Jing Gao, Yong-Jun Liu & Qing-Yun Diao
Scientific Reports 7, Article number: 15943 (2017)
doi:10.1038/s41598-017-16245-0

https://www.nature.com/articles/s41598-017-16245-0

Abstract
A sublethal concentration of imidacloprid can cause chronic toxicity in bees and can impact the behavior of honey bees. The nectar- and water-collecting, and climbing abilities of bees are crucial to the survival of the bees and the execution of responsibilities in bee colonies. Besides behavioral impact, data on the molecular mechanisms underlying the toxicity of imidacloprid, especially by the way of RNA-seq at the transcriptomic level, are limited. We treated Apis mellifera L. with sublethal concentrations of imidacloprid (0.1, 1 and 10 ppb) and determined the effect on behaviors and the transcriptomic changes. The sublethal concentrations of imidacloprid had a limited impact on the survival and syrup consumption of bees, but caused a significant increase in water consumption. Moreover, the climbing ability was significantly impaired by 10 ppb imidacloprid at 8 d. In the RNA-seq analysis, gene ontology (GO) term enrichment indicated a significant down-regulation of muscle-related genes, which might contribute to the impairment in climbing ability of bees. The enriched GO terms were attributed to the up-regulated ribosomal protein genes. Considering the ribosomal and extra-ribosomal functions of the ribosomal proteins, we hypothesized that imidacloprid also causes cell dysfunction. Our findings further enhance the understanding of imidacloprid sublethal toxicity.


----------



## BernhardHeuvel

http://www.db.zs-intern.de/uploads/1512373695-Birds.pdf

Imidacloprid and chlorpyrifos insecticides impair migratory ability in a seed-eating songbird 

Scientific Reports | 7: 15176 | DOI:10.1038/s41598-017-15446-x 1

Margaret L. Eng 1, Bridget J. M. Stutchbury2 & Christy A. Morrissey3,4


----------



## jonsl

*Re: Gene expression changes in honey bees induced by sublethal imidacloprid exposure*



BernhardHeuvel said:


> Not to me. But for some it is still a surprise that a highly toxic and systemic insecticide group is poisonous to bees in field application.
> 
> As Bayer said: chemical plus nature.


Probably not a suprise to these people, just an inconvenient fact.


----------



## Eduardo Gomes

*Re: Gene expression changes in honey bees induced by sublethal imidacloprid exposure*

"I was definitely surprised,” he said. “Fungicides have been largely overlooked.” However, a few lab-based studies have shown that fungicides can make nosema much worse in bees, probably by killing beneficial gut microbes."

https://www.theguardian.com/environ...-bee-declines-revealed?CMP=Share_iOSApp_Other


----------



## Buzz-kill

*Re: Gene expression changes in honey bees induced by sublethal imidacloprid exposure*



jonsl said:


> Probably not a suprise to these people, just an inconvenient fact.


“It is difficult to get a man to understand something, when his salary depends on his not understanding it.” -Upton Sinclair

Big Tobacco, Big Oil, Big Agriculture. All use the same playbook. Question and obfuscate and sow confusion in the public mind as to whether the science has proven their products are a detriment to civilization and at the same time buy the politicians.


----------



## clyderoad

*Re: Gene expression changes in honey bees induced by sublethal imidacloprid exposure*



Buzz-kill said:


> “It is difficult to get a man to understand something, when his salary depends on his not understanding it.” -Upton Sinclair
> 
> Big Tobacco, Big Oil, Big Agriculture. All use the same playbook. Question and obfuscate and sow confusion in the public mind as to whether the science has proven their products are a detriment to civilization and at the same time buy the politicians.


Good grief, Big Grief. 
Bees Buzzy, Bees. How are your Bees Buzzy? any problems with neonics or fungicides in your Bees?


----------



## Buzz-kill

*Re: Gene expression changes in honey bees induced by sublethal imidacloprid exposure*



clyderoad said:


> Good grief, Big Grief.
> Bees Buzzy, Bees. How are your Bees Buzzy? any problems with neonics or fungicides in your Bees?


Huh? I assume you have reading comprehension issues.


----------



## clyderoad

*Re: Gene expression changes in honey bees induced by sublethal imidacloprid exposure*



Buzz-kill said:


> Huh? I assume you have reading comprehension issues.


After reading your posts I think a reading comprehension issue might not be a bad thing.
But maybe the issue is yours, from post #1 in this thread:

"Use this thread to post Articles, Studies or Links that apply to the topic of CCD.
This will allow members to have all supporting data in one place.

This thread is NOT for discussion.
Post supporting data only. Discuss data in other threads."


----------



## BernhardHeuvel

*Re: Gene expression changes in honey bees induced by sublethal imidacloprid exposure*

*Effect of Sublethal Doses of Clothianidin and/or V. destructor on Honey Bee (Apis mellifera L.) Health, Behavior and Associated Gene Expression*

Autor:	Morfin Ramirez, Nuria
Department:	School of Environmental Sciences
Program:	Environmental Sciences
Advisor:	Guzman-Novoa, Ernesto

Abstract
Little is known about the effects of sublethal doses of neonicotinoids on honey bee (Apis mellifera L.) behaviors and mortality, and whether those effects are altered with parasitism by V. destructor. This study examined the effects of multiple exposures to field-realistic sublethal doses of clothianidin with and without V. destructor on adult bees and newly emerged bees treated as larvae. For adult exposure, memory retention decreased with each stressor alone, but weight and sugar consumption decreased only by the effect of V. destructor. For larval exposure, haemocyte counts increased with clothianidin but decreased with V. destructor, clothianidin reduced hygienic behavior and the number of foraging trips of the adults that emerged. Interactions between the stressors were observed as decreased weight of newly emerged bees with larval exposure, an increased mortality in adult bees, and a decreased intense grooming behavior with adult exposure. The relative expression of several immune and neural related honey bee genes showed an interaction between the stressors using two-way ANOVA in many cases. Also, the dose response of gene expression often revealed a non-linear pattern, implying hormesis, although hormesis was not detected for any of the biological measurements. For example, AmpUf68 expression in newly emerged bees showed an interaction between the stressors with a J-shaped dose response to clothianidin and no dose response to clothianidin plus V. destructor, while AmDef-2 expression in adults showed an interaction between stressors with an inverted U-shaped dose response to clothianidin and a sigmoidal dose response to clothianidin plus V. destructor. RNAseq analysis of bees with the highest sublethal doses of clothianidin with and without V. destructor showed no changes in the magnitude of expression but reduced numbers of differentially expressed genes with the combined stressors compared to each stressor alone. However, novel differentially expressed genes were also observed with the combined stressors. The combined stressors appeared to both change and inhibit the numbers of differentially expressed genes compared to each stressor alone. In general, clothianidin and V. destructor have different effects on bee health and behavior that was only rarely affected when combined, whereas gene expression mostly had reduced and unpredictable, rather than additive, effects with the combined stressors.

URI:	http://hdl.handle.net/10214/12547
https://atrium.lib.uoguelph.ca/xmlui/handle/10214/12547


----------



## BernhardHeuvel

*Re: Gene expression changes in honey bees induced by sublethal imidacloprid exposure*

European Food Safety Authority:
*Neonicotinoids: risks to bees confirmed*
http://www.efsa.europa.eu/en/press/news/180228


----------



## BernhardHeuvel

*Re: Gene expression changes in honey bees induced by sublethal imidacloprid exposure*

https://www.ncbi.nlm.nih.gov/pubmed/29499530

Sci Total Environ. 2018 Feb 27;630:487-494. doi: 10.1016/j.scitotenv.2018.02.258. [Epub ahead of print]

*Enhancement of chronic bee paralysis virus levels in honeybees acute exposed to imidacloprid: A Chinese case study.*

Diao Q1, Li B1, Zhao H2, Wu Y1, Guo R3, Dai P1, Chen D3, Wang Q1, Hou C4.

Abstract
Though honeybee populations have not yet been reported to be largely lost in China, many stressors that affect the health of honeybees have been confirmed. Honeybees inevitably come into contact with environmental stressors that are not intended to target honeybees, such as pesticides. Although large-scale losses of honeybee colonies are thought to be associated with viruses, these viruses usually lead to covert infections and to not cause acute damage if the bees do not encounter outside stressors. To reveal the potential relationship between acute pesticides and viruses, we applied different doses of imidacloprid to adult bees that were primarily infected with low levels (4.3×105 genome copies) of chronic bee paralysis virus (CBPV) to observe whether the acute oral toxicity of imidacloprid was able to elevate the level of CBPV. Here, we found that the titer of CBPV was significantly elevated in adult bees after 96h of acute treatment with imidacloprid at the highest dose 66.9ng/bee compared with other treatments and controls. Our study provides clear evidence that exposure to acute high doses of imidacloprid in honeybees persistently infected by CBPV can exert a remarkably negative effect on honeybee survival. These results imply that acute environmental stressors might be one of the major accelerators causing rapid viral replication, which may progress to cause mass proliferation and dissemination and lead to colony decline. The present study will be useful for better understanding the harm caused by this pesticide, especially regarding how honeybee tolerance to the viral infection might be altered by acute pesticide exposure.


----------



## little_john

*Re: Gene expression changes in honey bees induced by sublethal imidacloprid exposure*



Buzz-kill said:


> Big Tobacco, Big Oil, Big Agriculture. All use the same playbook. Question and obfuscate and sow confusion in the public mind as to whether *the science has proven* their products are a detriment to civilization and at the same time buy the politicians.


But science never gets to *prove* anything ...

Surprisingly to some, science does not deal in proof, in spite of the word being associated with science a whole lot more than perhaps it should be. https://rationalwiki.org/wiki/Proof

We can demonstrate, suggest, and convince ourselves that a scientific truth is valid. But proof? That's an impossibility for science. https://www.forbes.com/sites/startswithabang/2017/11/22/scientific-proof-is-a-myth/

While the phrase "scientific proof" is often used in the popular media, many scientists have argued that there is really no such thing. For example, Karl Popper once wrote that "In the empirical sciences, which alone can furnish us with information about the world we live in, proofs do not occur, if we mean by 'proof' an argument which establishes once and for ever the truth of a theory,". https://en.wikipedia.org/wiki/Scientific_evidence
LJ


----------



## 1102009

This thread is not for discussion:



Barry said:


> Use this thread to post Articles, Studies or Links that apply to the topic of CCD.
> This will allow members to have all supporting data in one place.
> 
> This thread is NOT for discussion.
> Post supporting data only. Discuss data in other threads.


----------



## johno

SiWolke, there has been discussion taking place in all 8 pages of this thread, even by yourself. Perhaps you would like only discussion favouring only your point of view? As a beekeeper surrounded by agriculture using neonics I generally have less than a 10% loss each year. However I had a small yard where I could not keep my colonies alive and one day while visiting that yard I saw a neighbor spraying in his vegetable garden so I wandered over there and asked what he was spraying with to which he replied Sevin, well I no longer keep bees there as I know why I could not keep them alive.
Now the moral of the story is Neonics could harm bees, it is an insecticide but some of the stuff the Ag sector used to use before Neonics was a lot worse and killed a lot more bees. And the story about proof, who needs proof when we have a consensus of scientists just like the consensus of scientists who believed the world was flat before Gallileo came along well you know that story. It appears that today the consensus of scientists is mainly a consensus of political science graduates.
Johno


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## little_john

SiWolKe said:


> This thread is not for discussion:


I wasn't 'discussing' - as a graduate in both Organic Chemistry and General Biology, I was simply correcting a false assumption. One which is frequently applied to the issues of CCD, Neonicotinoids - and many others currently related to beekeeping.
LJ


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## 1102009

johno said:


> SiWolke, there has been discussion taking place in all 8 pages of this thread, even by yourself.


Yes but I learned. I did not realize. Never again. It´s hard to find the links if you want to use them again if there is discussion between.

There are many other threads in this sub forum with discussion about the topics.

Little_john
that´s interesting, please post your links if they are contradiction

Bernhard, many thanks for keeping us updated.


----------



## psm1212

*Re: Gene expression changes in honey bees induced by sublethal imidacloprid exposure*



BernhardHeuvel said:


> Not to me. But for some it is still a surprise that a highly toxic and systemic insecticide group is poisonous to bees in field application.
> 
> As Bayer said: chemical plus nature.


Field application? Wouldn't field application be accomplished by nurse bees eating pollen and honey laden with neonics and then feeding the queen larva royal jelly? By "field application" are you referencing the Gajger, Sakač, & Gregorc study that you cite above? 

From the abstract "honey bee larvae *received a single treatment dose* (4.28 ng thiamethoxam/queen larva on the 4th day after larvae grafting in artificial queen cells), while the second honey bee queen rearing cycle *received a double treatment dose* (total of 8.56 ng thiamethoxam/queen larva on the 4th and 5th day after larvae grafting in artificial queen cells)."

I do not see how injecting thiamethoxam into a developing queen cell ever occurs in the "field."


----------



## BernhardHeuvel

*Re: Gene expression changes in honey bees induced by sublethal imidacloprid exposure*

*Influences of acephate and mixtures with other commonly used pesticides on honey bee (Apis mellifera) survival and detoxification enzyme activities*

Jianxiu Yaoa, Yu Cheng Zhua, John Adamczyk, Randall Luttrell
https://www.sciencedirect.com/science/article/pii/S1532045618300218
https://doi.org/10.1016/j.cbpc.2018.03.005

Abstract
Acephate (organophosphate) is frequently used to control piercing/sucking insects in field crops in southern United States, which may pose a risk to honey bees. In this study, toxicity of acephate (formulation Bracket®97) was examined in honey bees through feeding treatments with sublethal (pollen residue level: 0.168 mg/L) and median-lethal (LC50: 6.97 mg/L) concentrations. Results indicated that adult bees treated with acephate at residue concentration did not show significant increase in mortality, but esterase activity was significantly suppressed. Similarly, bees treated with binary mixtures of acephate with six formulated pesticides (all at residue dose) consistently showed lower esterase activity and body weight. Clothianidin, λ-cyhalothrin, oxamyl, tetraconazole, and chlorpyrifos may interact with acephate significantly to reduce body weight in treated bees. The dose response data (LC50: 6.97 mg/L) revealed a relatively higher tolerance to acephate in Stoneville bee population (USA) than populations elsewhere, although in general the population is still very sensitive to the organophosphate. In addition to killing 50% of the treated bees acephate (6.97 mg/L) inhibited 79.9%, 20.4%, and 29.4% of esterase, Glutathione S-transferase (GST), and acetylcholinesterase (AChE) activities, respectively, in survivors after feeding treatment for 48 h. However, P450 activity was elevated 20% in bees exposed to acephate for 48 h. Even though feeding on sublethal acephate did not kill honey bees directly, chronic toxicity to honey bee was noticeable in body weight loss and esterase suppression, and its potential risk of synergistic interactions with other formulated pesticides should not be ignored.


----------



## BernhardHeuvel

*Re: Gene expression changes in honey bees induced by sublethal imidacloprid exposure*

Have a quick look at: 

http://www.agrofog.com/brochures/bayer-premise200sc.pdf

That's an ad from the manufacturer of the pesticide. Read carefully what a neonic does. It boils down to: chemical plus nature. The rest is self-explaining.

Good luck to you all. Gonna be tough times, for both: bees and beekeepers that make a living from their bees.


----------



## jonsl

*Re: Gene expression changes in honey bees induced by sublethal imidacloprid exposure*



BernhardHeuvel said:


> Have a quick look at:
> 
> http://www.agrofog.com/brochures/bayer-premise200sc.pdf
> 
> That's an ad from the manufacturer of the pesticide. Read carefully what a neonic does. It boils down to: chemical plus nature. The rest is self-explaining.
> 
> Good luck to you all. Gonna be tough times, for both: bees and beekeepers that make a living from their bees.


The one in the link is of particular concern: half life of 355 days.


----------



## Varroa Apiary

*Re: Gene expression changes in honey bees induced by sublethal imidacloprid exposure*

Carreck, Norman and Ratnieks, Francis (2014) The dose makes the poison: have "field realistic" rates of exposure of bees to neonicotinoid insecticides been overestimated in laboratory studies? Journal of Apicultural Research, 53 (5). pp. 607-614. ISSN 0021-8839
|"Recent laboratory based studies have demonstrated adverse sub-lethal effects of neonicotinoid insecticides on honey bees and bumble bees, and these studies have been influential in leading to a European Union moratorium on the use of three neonicotinoids, clothianidin, imidacloprid, and thiamethoxam on "bee attractive" crops. Yet so far, these same effects have not been observed in field studies. Here we review the three key dosage factors (concentration, duration and choice) relevant to field conditions, and conclude that these have probably been over estimated in many laboratory based studies."
https://www.tandfonline.com/doi/abs/10.3896/IBRA.1.53.5.08

Three years of banning neonicotinoid insecticides based on sub‐lethal effects: can we expect to see effects on bees?
Tjeerd Blacquière Jozef JM van der Steen
First published: 03 April 2017 https://doi.org/10.1002/ps.4583 Cited by: 4
Although no direct relationship between colony losses and the decline of numbers of honey bee colonies could be shown (see section 4), losses of a high number of colonies over winter are considered to be a problem in itself for beekeepers, and have stimulated the foundation of the Coloss network.14 So could honey bee colony losses experienced by beekeepers be attributed to the use of neonicotinoids? Smith et al.11 point out that the evidence is not strong for the case. Even a very extensive 4‐year monitoring project set‐up with the intention to shed light on the possible factors involved in honey bee colony losses, and specifically focusing on residues of chemicals,19 was not able to show any relationship with these, but did show effects of infestation with the Varroa mite, some viruses and the age of the queen: actually all being part of the management choices of the beekeeper, a decisive factor often overlooked.
https://onlinelibrary.wiley.com/doi/full/10.1002/ps.4583

Environ Health Perspect; DOI:10.1289/EHP515
Effects of Neonicotinoid Pesticide Exposure on Human Health: A Systematic Review
Andria M. Cimino,1 Abee L. Boyles,2 Kristina A. Thayer,2 and Melissa J. Perry1
To the authors’ knowledge, this is the first systematic review of the literature on human health effects of neonicotinoids. As reviewed here, four studies reported low rates of adverse health effects from acute neonic exposure. Even the most severe outcomes, including two fatalities, may have been mediated by other factors (age, underlying health conditions, undetected coexposures). The acute poisoning studies did, however, elucidate clinical findings important for the diagnosis and treatment of acute neonic exposures, including a better understanding of neonic toxicokinetics in humans. The other four studies reported associations between chronic neonic exposure and adverse developmental outcomes or a symptom cluster including neurological effects. The findings of animal studies support the biological plausibility for such associations (Abou-Donia et al. 2008; Gibbons et al. 2015; Gu et al. 2013; Kimura-Kuroda et al. 2012; Li et al. 2011; Mason et al. 2013; Tomizawa 2004).

Although the studies in this review represent an important contribution to the literature, particularly given the lack of any general population chronic exposure studies prior to 2014, there remains a paucity of data on neonic exposure and human health. Given the widespread use of neonics in agriculture and household products and its increasing detection in U.S. food and water, more studies on the human health effects of chronic (non-acute) neonic exposure are needed.
https://ehp.niehs.nih.gov/ehp515/


----------



## 1102009

*Re: Gene expression changes in honey bees induced by sublethal imidacloprid exposure*

Glyphosate perturbs the gut microbiota of honey bees.

http://www.pnas.org/content/early/2018/09/18/1803880115


----------



## 1102009

*Re: Gene expression changes in honey bees induced by sublethal imidacloprid exposure*

Effects of sublethal doses of glyphosate on honeybee navigation

http://jeb.biologists.org/content/218/17/2799


----------



## 1102009

*Re: Gene expression changes in honey bees induced by sublethal imidacloprid exposure*

http://www.pnas.org/content/pnas/115/41/10305.full.pdf


----------



## Eduardo Gomes

*Re: Gene expression changes in honey bees induced by sublethal imidacloprid exposure*

https://thoughtscapism.com/2018/06/11/no-glyphosate-is-not-a-threat-to-bees/


----------



## crofter

*Re: Gene expression changes in honey bees induced by sublethal imidacloprid exposure*



Eduardo Gomes said:


> https://thoughtscapism.com/2018/06/11/no-glyphosate-is-not-a-threat-to-bees/


Thanks, Eduardo; 

A good example of very weak research designed to support an ideological opinion.

People susceptible to this kind of persuasion are generally not inclined to dig deep enough to spot the issues that could suggest very different causes for the observations. 

Precise science is not needed to appeal to emotions!


----------



## 1102009

*Re: Gene expression changes in honey bees induced by sublethal imidacloprid exposure*

Scientists are scientists.
Look at the dates.

I´m very sad that links are still discussed here.
Well, Barry left and moderation changed.


Threads
148,053
Posts
1,578,298
Members
34,726
*Active Members
1,984 *

says it all.


----------



## little_john

*Re: Gene expression changes in honey bees induced by sublethal imidacloprid exposure*



SiWolKe said:


> Threads
> 148,053
> Posts
> 1,578,298
> Members
> 34,726
> *Active Members
> 1,984 *
> 
> says it all.


Hello Sibylle - I may be having one of my really 'thick' days today - but what do you mean by "says it all" ? You've flagged-up nearly two thousand active members, so I guess that's part of the point you're making ... 

Maybe I've missed something important within the last few posts ?
'best
LJ


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## Fivej

*Re: Gene expression changes in honey bees induced by sublethal imidacloprid exposure*

Neonic Study On Bumblebees: https://www.npr.org/2018/11/09/6656...ees-see-harmful-effects-of-common-insecticide


----------



## BernhardHeuvel

*Re: Gene expression changes in honey bees induced by sublethal imidacloprid exposure*

*Effects of clothianidin exposure on semen parameters of honey bee drones.*
Abstract : Many problems have been reported on honey bees colonies including fertility problems of queens resulted in production failure. Pesticides can be the cause of this failure in connection with the quality of sperm drone. Thus, the aim of this study was to assess the influence of exposure to syrup contaminated with clothianidin at 0.1 µg/L on semen parameters of drones. Results showed a significant decrease of semen volume and sperm concentration and an increase in sperm mortality rate. As for the energetic state, clothianidin increased cell redox potential, the ATP content of spermatozoa as well as the lactate dehydrogenase activity (LDH). It was concluded that exposure to clothianidin during the sexual maturity of drones could affect the semen quality.
https://www.cabdirect.org/cabdirect/abstract/20193066560

*Parameterization and sensitivity analysis of a honey bee colony dynamics model for neonicotinoid exposure events using Markov Chain Monte Carlo methods*
Abstract
Honey bee (Apis mellifera) colony losses have increased in recent decades in both Europe and North America. While multiple stressors to honey bee colonies appear to be driving this decline (including disease, nutrition, genetics), direct exposure to pesticides has been identified as a factor leading to increased bee declines. The simulation model VarroaPop is currently being modified by the USDA and USEPA to predict honey bee hive dynamics in response to pesticide exposure. However, applying this model to pesticides is complicated due to a lack of parameterization information from the supporting literature for many variables, especially those related to in-hive pesticide dynamics. Here, we utilize data from a field study which measured residues of several neonicotinoid insecticides in pollen and tracked population dynamics of exposed hives to improve our estimation of colony simulation model parameters relevant to VarroaPop and the new model components related to pesticides. We use Markov Chain Monte Carlo methods to sample the probability distribution of model parameters and examine the likelihood of each parameter combination, given the field-derived population data. Through this procedure, we obtain posterior distributions which represent the most likely parameter values given a realistic neonicotinoid exposure scenario. We use these pesticide-optimized parameter distributions to run a global sensitivity analysis for the updated posteriors in order to contrast with a sensitivity analysis based on the priors. This helps determine what factors are most important in driving hive success or failure following exposure events.
https://scholarsarchive.byu.edu/iemssconference/2018/Stream-G/7/

*Assessing the Impact of the Conservation Reserve Program on Honey Bee Health*
Key Findings (2014–18)
• More than one-sixth of all honey bee yards in North Dakota and South Dakota (the top two honey-producing States) meet the key foraging requirements of honey bees just based on the existence of CRP grasslands alone. Thus, the CRP plays a considerable role in supporting the carrying capacity of honey bee colonies in this region (Otto and others, 2018).
• Honey bee colonies in areas surrounded by grassland, such as those enrolled in the CRP, are 10–15 percent larger than colonies kept in areas surrounded by mostly row crops. A USGS study determined that a bee yard surrounded by mostly grassland can yield an extra $4,100 in annual revenue to a beekeeper compared to a bee yard surrounded by mostly row crop. A beekeeper who manages several hundred bee yards may incur sizable economic gains through nearby CRP and other conservation grass- lands (Smart and others, 2018).
• The USGS led the first large-scale assessment of native bees on CRP grassland that documented what native bees were present on CRP grasslands and what flowers they used (Otto and others, 2017).
• The USGS developed a genetic sequencing strategy to rapidly quantify pollen grains collected from bees (Smart and others, 2017). This novel technique allowed the USGS to determine what flowering plants were impor- tant bee food. Knowing what flowers constitute good
bee food is an important step in improving the cost- effectiveness of the CRP.
• The USGS launched the “Pollinator Library,” a decision- support tool that can be used by USDA staff for evalu- ating seeding mixes for the CRP (https://www.npwrc. usgs.gov/pollinator/). This website provides users with information on which plants are favored by honey bees and native bees. USGS scientists published a paper demonstrating how USDA staff can use the Pollinator Library to evaluate the cost-effectiveness of CRP seeding mixes (Otto and others, 2017).
https://pubs.usgs.gov/fs/2018/3082/fs20183082.pdf

*Hazard of a neonicotinoid insecticide on the homing flight of the honeybee depends on climatic conditions and Varroa infestation*
Abstract
The paradigm for all toxicological bioassays in the risk assessment of pesticide registration reflects the principle that experimental conditions should be controlled to avoid any other factors that may affect the endpoint measures. As honeybee colonies can be frequently exposed to bio-aggressors in real conditions, often concomitantly with pesticides, co-exposure to pesticide/bio-aggressors is becoming a concern for regulatory authorities. We investigated the effects of the neonicotinoid insecticide thiamethoxam on the homingperformances of foragers emerging from colonies differentiated by health status (infestation with Varroa destructor mites, microsporidian parasite Nosema spp. and Deformed Wing Virus). We designed a homing test that has been recently identified to fill a regulatory gap in the field evaluations of sublethal doses of pesticides before their registration. We also assessed the effect of temperature as an environmental factor. Our results showed that the Varroa mite exacerbates homing failure (HF) caused by the insecticide, whereas high temperatures reduce insecticide-induced HF. Through an analytical Effective Dose (ED) approach, predictive modeling results showed that, for instance, ED level of an uninfested colony, can be divided by 3.3 when the colony is infested by 5 Varroa mites per 100 bees and at a temperature of 24 °C. Our results suggest that the health status of honeybee colonies and climatic context should be targeted for a thorough risk assessment.
https://www.sciencedirect.com/science/article/pii/S0045653519303534


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## BernhardHeuvel

*Alterations in honey bee gut microorganisms*
Honey bees are associated with gut microorganisms essential for their nutrition and health. The composition of the microbial community can be used as a biological health indicator and is characterized using biomarker fatty acids. Commonly, gut microorganisms are exposed to pathogens and to an array of chemical and biological pest control methods.


We found a strong negative effect on microbial gut community composition when exposed to the bee pest control chemicals oxytetracycline, oxalic acid and imidacloprid, and when inoculated with the bee pest Nosema spp. and the potential bee pest biocontrol agent Lactobacillus plantarum. Results from the in vitro test with bee pest chemicals showed a differential response of Lactobacillus spp. At the community level, some taxonomic groups were more affected depending on treatment, but sharp changes in the microbial structure were caused by compounds generally considered as bee safe.


Our results show that pests such as Nosema spp. and pest control methods alter the composition of bee gut microorganisms, which may have severe consequences for pathogen defense, physiology and general honey bee health. In addition, L. plantarum has potential as a biocontrol agent against Nosema spp. © 2018 Society of Chemical Industry
https://www.ingentaconnect.com/content/jws/ps/2019/00000075/00000003/art00035


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## BernhardHeuvel

*Sublethal Effects of the Insecticide Pyrifluquinazon on the European Honey Bee (Hymenoptera: Apidae)*
James M Wilson Troy D Anderson Thomas P Kuhar
Journal of Economic Entomology, toz014, https://doi.org/10.1093/jee/toz014

Abstract
Pyrifluquinazon (PQZ) is an Insecticide Resistance Action Committee (IRAC) Group 9 insecticide that has recently been registered for use in the United States for control of soft-bodied sucking insect pests. Although it has been classified as practically nontoxic to honey bees, Apis mellifera L. (Hymenoptera: Apidae), based on acute contact bioassays, additional information on sublethal effects of this insecticide on honey bees is lacking. Using a combination of laboratory assays with video movement tracking software and near-field evaluations of colonies foraging in a high-tunnel experiment, we determined that, when fed PQZ at a concentration of 84 mg active ingredient (ai)/liter (= ppm) in sugar water, a reduction in overall movement by the foraging worker bees was observed. However, when provided with honey reserves in the hive, honey bees rejected the PQZ-treated sugar water. These results indicate that, if ingested at levels of 84 mg ai/liter, PQZ could have a negative effect on honey bee behavior; however, honey bee workers appear to be able to detect the presence of PQZ in their food and reject it.
https://academic.oup.com/jee/advance-article-abstract/doi/10.1093/jee/toz014/5320965


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## BernhardHeuvel

*Honey Bee Exposure to Pesticides: A Four-Year Nationwide Study*
Abstract
Pollinators, including honey bees, are responsible for the successful reproduction of more than 87% of flowering plant species: they are thus vital to ecosystem health and agricultural services world-wide. To investigate honey bee exposure to pesticides, 168 pollen samples and 142 wax comb samples were collected from colonies within six stationary apiaries in six U.S. states. These samples were analyzed for evidence of pesticides. Samples were taken bi-weekly when each colony was active. Each apiary included thirty colonies, of which five randomly chosen colonies in each apiary were sampled for pollen. The pollen samples were separately pooled by apiary. There were a total of 714 detections in the collected pollen and 1008 detections in collected wax. A total of 91 different compounds were detected: of these, 79 different pesticides and metabolites were observed in the pollen and 56 were observed in the wax. In all years, insecticides were detected more frequently than were fungicides or herbicides: one third of the detected pesticides were found only in pollen. The mean (standard deviation (SD)) number of detections per pooled pollen sample varied by location from 1.1 (1.1) to 8.7 (2.1). Ten different modes of action were found across all four years and nine additional modes of action occurred in only one year. If synergy in toxicological response is a function of simultaneous occurrence of multiple distinct modes of action, then a high frequency of potential synergies was found in pollen and wax-comb samples. Because only pooled pollen samples were obtained from each apiary, and these from only five colonies per apiary per year, more data are needed to adequately evaluate the differences in pesticide exposure risk to honey bees among colonies in the same apiary and by year and location. 
https://www.mdpi.com/2075-4450/10/1/13


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## BernhardHeuvel

Insects 2019, 10(1), 19; https://doi.org/10.3390/insects10010019
*Initial Exposure of Wax Foundation to Agrochemicals Causes Negligible Effects on the Growth and Winter Survival of Incipient Honey Bee (Apis mellifera) Colonies*
Abstract
Widespread use of agrochemicals in the U.S. has led to nearly universal contamination of beeswax in honey bee hives. The most commonly found agrochemicals in wax include beekeeper-applied miticides containing tau-fluvalinate, coumaphos, or amitraz, and field-applied pesticides containing chlorothalonil or chlorpyrifos. Wax contaminated with these pesticides negatively affects the reproductive quality of queens and drones. However, the synergistic effects of these pesticides on the growth and survival of incipient colonies remain understudied. We established new colonies using frames with wax foundation that was pesticide free or contaminated with field-relevant concentrations of amitraz alone, a combination of tau-fluvalinate and coumaphos, or a combination of chlorothalonil and chlorpyrifos. Colony growth was assessed by estimating comb and brood production, food storage, and adult bee population during a colony’s first season. We also measured colony overwintering survival. We found no significant differences in colony growth or survivorship between colonies established on pesticide-free vs. pesticide-laden wax foundation. However, colonies that had Varroa destructor levels above 3% in the fall were more likely to die over winter than those with levels below this threshold, indicating that high Varroa infestation in the fall played a more important role than initial pesticide exposure of wax foundation in the winter survival of newly established colonies.
https://www.mdpi.com/2075-4450/10/1/19


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## BernhardHeuvel

*Combined Toxicity of Insecticides and Fungicides Applied to California Almond Orchards to Honey Bee Larvae and Adults*
Insects 2019, 10(1), 20; https://doi.org/10.3390/insects10010020
Abstract
Beekeepers providing pollination services for California almond orchards have reported observing dead or malformed brood during and immediately after almond bloom—effects that they attribute to pesticide exposure. The objective of this study was to test commonly used insecticides and fungicides during almond bloom on honey bee larval development in a laboratory bioassay. In vitro rearing of worker honey bee larvae was performed to test the effect of three insecticides (chlorantraniliprole, diflubenzuron, and methoxyfenozide) and three fungicides (propiconazole, iprodione, and a mixture of boscalid-pyraclostrobin), applied alone or in insecticide-fungicide combinations, on larval development. Young worker larvae were fed diets contaminated with active ingredients at concentration ratios simulating a tank-mix at the maximum label rate. Overall, larvae receiving insecticide and insecticide-fungicide combinations were less likely to survive to adulthood when compared to the control or fungicide-only treatments. The insecticide chlorantraniliprole increased larval mortality when combined with the fungicides propiconazole or iprodione, but not alone; the chlorantraniliprole-propiconazole combination was also found to be highly toxic to adult workers treated topically. Diflubenzuron generally increased larval mortality, but no synergistic effect was observed when combined with fungicides. Neither methoxyfenozide nor any methoxyfenozide-fungicide combination increased mortality. Exposure to insecticides applied during almond bloom has the potential to harm honey bees and this effect may, in certain instances, be more damaging when insecticides are applied in combination with fungicides
https://www.mdpi.com/2075-4450/10/1/20


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## AHudd

BernhardHeuvel said:


> *Alterations in honey bee gut microorganisms*
> Honey bees are associated with gut microorganisms essential for their nutrition and health. The composition of the microbial community can be used as a biological health indicator and is characterized using biomarker fatty acids. Commonly, gut microorganisms are exposed to pathogens and to an array of chemical and biological pest control methods.
> 
> 
> We found a strong negative effect on microbial gut community composition when exposed to the bee pest control chemicals oxytetracycline, oxalic acid and imidacloprid, and when inoculated with the bee pest Nosema spp
> https://www.ingentaconnect.com/content/jws/ps/2019/00000075/00000003/art00035


I wonder if that is dribbled or vaporized OA. I read the link but I didn't see which.

Alex


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## BernhardHeuvel

Dribbled. Don't think sublimated oxalic acid do the same harm, since the bees don't lick it. It is known, that damage to the midgut by oxalic acid increases as the sugar content increases in the dribble solution. No sugar, no lick.


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## AHudd

BernhardHeuvel said:


> Dribbled. Don't think sublimated oxalic acid do the same harm, since the bees don't lick it. It is known, that damage to the midgut by oxalic acid increases as the sugar content increases in the dribble solution. No sugar, no lick.


Thanks for the response. That is what I was thinking/hoping.

Alex


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## BernhardHeuvel

*The novel pesticide flupyradifurone (Sivanto) affects honeybee motor abilities*

Abstract
Honeybees and other pollinators are threatened by changing landscapes and pesticides resulting from intensified agriculture. In 2018 the European Union prohibited the outdoor use of three neonicotinoid insecticides due to concerns about pollinators. A new pesticide by the name of “Sivanto” was recently released by Bayer AG. Its active ingredient flupyradifurone binds to the nicotinic acetylcholine receptor (AchR) in the honeybee brain, similar to neonicotinoids. Nevertheless, flupyradifurone is assumed to be harmless for honeybees and can even be applied on flowering crops. So far, only little has been known about sublethal effects of flupyradifurone on honeybees. Intact motor functions are decisive for numerous behaviors including foraging and dancing. We therefore selected a motor assay to investigate in how far sublethal doses of this pesticide affect behavior in young summer and long-lived winter honeybees. Our results demonstrate that flupyradifurone (830 µmol/l) can evoke motor disabilities and disturb normal motor behavior after a single oral administration (1.2 µg/bee). These effects are stronger in long-lived winter bees than in young summer bees. After offering an equal amount of pesticide (1.0–1.75 µg) continuously over 24 h with food the observed effects are slighter. For comparisons we repeated our experiments with the neonicotinoid imidacloprid. Intriguingly, the alterations in behavior induced by this pesticide (4 ng/bee) were different and longer-lasting compared to flupyradifurone, even though both substances bind to nicotinic acetylcholine receptors.
https://link.springer.com/article/10.1007/s10646-019-02028-y
https://doi.org/10.1007/s10646-019-02028-y


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## BernhardHeuvel

*DETECTION OF THE PESTICIDE IMIDACLOPRID IN HONEY*

K. E. Burkin

Abstract: Studies have been carried out to determine the imidacloprid pesticide content of the neonicotinoid group in honey, including sample preparation and subsequent analysis by high-performance liquid chromatography on a chromatograph with a UV-detector. The optimum conditions providing the maximum extraction of imidacloprid from honey are selected.

http://tipkadpo.ru/data/uploads/nauka/konferencii/2018/_2018__.pdf#page=83


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## BernhardHeuvel

*Impact of acute oral exposure to thiamethoxam on the homing, flight, learning acquisition and short‐term retention of Apis cerana*

Changsheng Ma Yongkui Zhang Jia Sun Muhammad Imran Huipeng Yang Jie Wu … other authors /et al.
First published: 18 March 2019 
https://doi.org/10.1002/ps.5411
https://onlinelibrary.wiley.com/doi/abs/10.1002/ps.5411

Abstract
BACKGROUND

Thiamethoxam (TMX) represents the second‐generation of neonicotinoids that has been widely applied in agricultural activities, while how TMX alters the behavior of Apis cerana, an important native honey bee species in China, is not clear. We carried out three independent experiments to study the impact of acute oral treatment of 20 μl TMX at concentrations of 2.4 ppb (0.048 ng/bee) and 10 ppb (0.2 ng/bee) on the homing, flight, learning acquisition and short‐term retention ability of A. cerana. The homing ability was assessed by the catch‐and‐release method, the flight ability was assessed by flight mills, and the learning acquisition and short‐term retention were evaluated by the proboscis extension response method.

RESULTS

When treated with 10 ppb of TMX, bees had a significantly higher average homing time, mean flight velocity, flying distance, and flying duration than the control, whereas 2.4 ppb concentration did not cause any significant effect on homing or flight ability. Bees treated with either 2.4 ppb or 10 ppb TMX had significantly lower learning acquisition and short‐term retention ability.

CONCLUSION

Results suggest that acute oral exposure to 10 ppb of TMX altered the short distance homing time, flight ability, and learning acquisition and short‐term retention ability. Our study also highlights the concern that acute oral exposure to a low concentration of 2.4 ppb could have consequences on the behavior of A. creana. Those multiple sublethal alterations on A. cerana's behavior indicate that TMX are likely causing complex, but negative consequences on bee health in the field.


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## BernhardHeuvel

*Effects of sublethal doses of clothianidin and/or V. destructor on honey bee (Apis mellifera) self-grooming behavior and associated gene expression*

Nuria Morfin, Paul H. Goodwin, Greg. J. Hunt & Ernesto Guzman-Novoa 

Scientific Reportsvolume 9, Article number: 5196 (2019)


Abstract

Little is known about the combined effects of stressors on social immunity of honey bees (Apis mellifera) and related gene expression. The interaction between sublethal doses of a neurotoxin, clothianidin, and the ectoparasite, Varroa destructor, was examined by measuring differentially expressed genes (DEGs) in brains, deformed wing virus (DWV) and the proportion and intensity of self-grooming. Evidence for an interaction was observed between the stressors in a reduction in the proportion of intense groomers. Only the lowest dose of clothianidin alone reduced the proportion of self-groomers and increased DWV levels. V. destructor shared a higher proportion of DEGs with the combined stressors compared to clothianidin, indicating that the effects of V. destructor were more pervasive than those of clothianidin when they were combined. The number of up-regulated DEGs were reduced with the combined stressors compared to clothianidin alone, suggesting an interference with the impacts of clothianidin. Clothianidin and V. destructor affected DEGs from different biological pathways but shared impacts on pathways related to neurodegenerative disorders, like Alzheimer’s, which could be related to neurological dysfunction and may explain their negative impacts on grooming. This study shows that the combination of clothianidin and V. destructor resulted in a complex and non-additive interaction.

...


Discussion

This study examined the effect of two stressors, sublethal doses of clothianidin and V. destructor on gene expression in the brain and increases in a mite-transmitted virus, DWV, to better understand how these two stressors act separately and together to affect honey bees on aspects of neural activity related to self-grooming behavior. Among the two stressors, clothianidin alone had a greater effect than V. destructor alone to reduce self-grooming behavior, based on the proportion of bees that self-groomed and the proportion of bees that self-groomed intensively, as well as the number of up-regulated DEGs. However, the results with the combination of V. destructor with clothianidin showed that combining each stressor did not result in a simple additive effect in the number of DEGs or the KEGG pathways associated with them.

One unexpected result of this study was that both DWV levels and the proportion of bees that self-groomed were affected only by the lowest dose of clothianidin without V. destructor. This may indicate that effects of DWV and grooming could be linked in some manner. Unexpectedly, higher doses of clothianidin in absence of mites, or the same dose of clothianidin combined with V. destructor, did not have the same impacts, indicating that there was a relatively specific impact from the lowest dose of clothianidin tested. Hormesis occurs when there is a beneficial biological response to a low exposure to a stressor28. In this case, a beneficial response did not occur at the lowest dose, but the effect could be similar to hormesis. At the lowest clothianidin dose, it would still bind to nicotinic acetylcholine receptors of the neural cells resulting in neural stimulation but may not overstimulate it to the level of toxic doses that block receptors impeding the action of the neurotransmitter ACh29. At very low doses, clothianidin stimulation of the central nervous system may just be sufficient to somehow interfere with self-grooming and resistance to a virus. While these results imply hormesis, future research should investigate more sublethal doses of clothianidin to confirm this by determining the range of doses that results in these effects.

...


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## BernhardHeuvel

*Effects of three common pesticides on survival, food consumption and midgut bacterial communities of adult workers Apis cerana and Apis mellifera*

YangYangaShilongMaabZhenxiongYancFengLiudQingyunDiaoaPingliDaia

https://doi.org/10.1016/j.envpol.2019.03.077

Highlights
•
Apis mellifera was more sensitive to chlorpyrifos and dimethoate than Apis cerana.

•
Amitraz at a concentration of 7.8 mg/L impacted Apis cerana survival.

•
Chlorpyrifos at a concentration of 3.0 mg/L impacted Apis cerana survival.

•
Dimethoate at a concentration of 1.0 mg/L impacted both Apis mellifera and Apis cerana survival.

•
There was a significant difference in microbiota species richness among treatments at day 15.


Abstract
The acute and chronic toxicity of 3 common pesticides, namely, amitraz, chlorpyrifos and dimethoate, were tested in Apis mellifera and Apis cerana. Acute oral toxicity LC50 values were calculated after 24 h of exposure to contaminated syrup, and chronic toxicity was tested after 15 days of exposure to 2 sublethal concentrations of pesticides. The toxicity of the tested pesticides to A. mellifera and A. cerana decreased in the order of dimethoate > chlorpyrifos > amitraz. A. mellifera was slightly more sensitive to chlorpyrifos and dimethoate than A. cerana, while A. cerana was more sensitive to amitraz than A. mellifera. Chronic toxicity tests showed that 1.0 mg/L dimethoate reduced the survival of the two bee species and the food consumption of A. mellifera, while 1.0 mg/L amitraz and 1.0 mg/L chlorpyrifos did not affect the survival or food consumption of the two bee species. The treatment of syrup with amitraz at a concentration equal to 1/10th of the LC50 value did not affect the survival of or diet consumption by A. mellifera and A. cerana; however, chlorpyrifos and dimethoate at concentrations equal to 1/10th of their respective LC50 values affected the survival of A. cerana. Furthermore, intestinal bacterial communities were identified using high-throughput sequencing targeting the V3V4 regions of the 16S rDNA gene. All major honey bee intestinal bacterial phyla, including Proteobacteria (62.84%), Firmicutes (34.04%), and Bacteroidetes (2.02%), were detected. There was a significant difference in the microbiota species richness of the two species after 15 days; however, after 30 days, no significant differences were found in the species diversity and richness between A. cerana and A. mellifera exposed to 1.0 mg/L amitraz and 1.0 mg/L chlorpyrifos. Overall, our results confirm that acute toxicity values are valuable for evaluating the chronic toxicity of these pesticides to honey bees.


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## BernhardHeuvel

*Sub-lethal concentrations of neonicotinoid insecticides at the field level affect neg*

*Sub-lethal concentrations of neonicotinoid insecticides at the field level affect negatively honey yield: Evidence from a 6-year survey of Greek apiaries*

Robert G. Chambers, Konstantinos Chatzimichael, Vangelis Tzouvelekas

Abstract
The threats posed by neonicotinoid insecticides to bee populations have been the focus of considerable research. Previous work has shed new light on the effects of neonicotinoids on bees by uncovering pathways through which neonicotinoids affect bee population dynamics and the potential interactions they have with exogenous stressors. Yet, little is known about whether these effects translate in a field-relevant setting to substantial losses in honey yields for commercial beekeepers. Here, we used data from a 6-year survey of 60 apiaries in Greece and economic modelling to assess at the field level the effects of neonicotinoid insecticides on honey production. Based on production function estimates, we found that sub-lethal concentrations of two widely used neonicotinoid insecticides (imidacloprid and thiamethoxam) detected in the nectar of flowers resulted in substantial losses in honey production for commercial beekeepers in our sample. By simulating a scenario with ideal pathogenic and environmental conditions, we found that the magnitude of the neonicotinoid effects decreases significantly under ideal conditions providing evidence for possible synergies at the field between neonicotinoids and environmental and pathogenic factors. Moreover, in a replicated study with grouped apiaries, we found evidence that the marginal effects of neonicotinoids on honey production may vary across apiaries facing different conditions.

https://economics.soc.uoc.gr/wpa/docs/1901.pdf


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## SuiGeneris

BernhardHeuvel said:


> *Effects of three common pesticides on survival, food consumption and midgut bacterial communities of adult workers Apis cerana and Apis mellifera*
> 
> YangYangaShilongMaabZhenxiongYancFengLiudQingyunDiaoaPingliDaia
> 
> https://doi.org/10.1016/j.envpol.2019.03.077
> 
> Highlights
> •
> Apis mellifera was more sensitive to chlorpyrifos and dimethoate than Apis cerana.
> 
> •
> Amitraz at a concentration of 7.8 mg/L impacted Apis cerana survival.
> 
> •
> Chlorpyrifos at a concentration of 3.0 mg/L impacted Apis cerana survival.
> 
> •
> Dimethoate at a concentration of 1.0 mg/L impacted both Apis mellifera and Apis cerana survival.
> 
> •
> There was a significant difference in microbiota species richness among treatments at day 15.


There's a hugely flawed study if I ever saw one! The concentrations of all of those is around the tank concentration used in the field. So even if the bees drank right from the sprayer on the tractor, they'd probably "bee" OK. For example 3.0 mg/L of chlorpyrifos = 0.3%; when you prepare chlorpyrifos for spraying you dilute it to 0.1% to 0.5% in the tank :lpf:


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## BernhardHeuvel

So all is ok. Back to sleep.


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## SuiGeneris

I never said that, but ****** science is ****** science. Grist for the propaganda machine, I guess, but no evidentiary value.


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## BernhardHeuvel

Cauia, Eliza & Siceanu, & Vișan, & Colța, & Spulber,. (2020). *Monitoring the Field-Realistic Exposure of Honeybee Colonies to Neonicotinoids by An Integrative Approach: A Case Study in Romania.* Diversity. 12. 24. 10.3390/d12010024. 

Abstract
Honeybees (Apis mellifera L.) are excellent biosensors that can be managed to collect valuable information about environmental contamination. The main objective of the present study was to design and apply an integrative protocol to monitor honeybee colony activity and sample collection by using electronic technologies combined with classical methods in order to evaluate the exposure of honeybees to the neonicotinoids that are used in melliferous intensive crops. The monitored honeybee colonies were especially prepared and equipped to maximize their chances to collect representative samples in order to express, as well as possible, the pesticide residues that existed in the targeted crops. The samples of honey, pollen and honeybees were collected, preserved and prepared to fulfill the required quality and quantity criteria of the accredited laboratories. In total, a set of fifty samples was collected from fields, located in different areas of intensive agriculture in Romania, and was analyzed for five neonicotinoids. The obtained results show that 48% of the total analyzed samples (n = 50) contained one or more detected or quantified neonicotinoid residues. The main conclusion is that the proposed approach for sample collection and preparation could improve the evaluation methodologies for analyzing honeybees’ exposure to pesticides.
https://www.researchgate.net/public..._Integrative_Approach_A_Case_Study_in_Romania



Comment: we found, that freezing bees greatly improved in finding the residues in the lab. Also collect poisoned bees that are still alive. The process of rotting in the bee greatly destroys residues.


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## BernhardHeuvel

*Honeybees fail to discriminate floral scents in a complex learning task after consuming a neonicotinoid pesticide*

Abstract

Neonicotinoids are pesticides used to protect crops but with known secondary influences at sublethal doses on bees. Honeybees use their sense of smell to identify the queen and nestmates, to signal danger and to distinguish flowers during foraging. Few behavioural studies to date have examined how neonicotinoid pesticides affect the ability of bees to distinguish odours. Here, we use a differential learning task to test how neonicotinoid exposure affects learning, memory, and olfactory perception in foraging-age honeybees. Bees fed with thiamethoxam could not perform differential learning and could not distinguish odours during short and long-term memory tests. Our data indicate that thiamethoxam directly impacts the cognitive processes involved in working memory required during differential olfactory learning. Using a combination of behavioural assays, we also identified that thiamethoxam has a direct impact on the olfactory perception of similar odours. Honeybees fed with other neonicotinoids (clothianidin, imidacloprid, dinotefuran) performed the differential learning task, but at a slower rate than the control. These bees could also distinguish the odours. Our data are the first to show that neonicotinoids have compound specific effects on the ability of bees to perform a complex olfactory learning task. Deficits in decision-making caused by thiamethoxam exposure could be more harmful than other neonicotinoids, leading to inefficient foraging and a reduced ability to identify nest mates.

Honeybees fail to discriminate floral scents in a complex learning task after consuming a neonicotinoid pesticide
Julie A. Mustard, Anne Gott, Jennifer Scott, Nancy L. Chavarria, Geraldine A. Wright
Journal of Experimental Biology 2020 : jeb.217174 doi: 10.1242/jeb.217174 Published 6 February 2020
https://jeb.biologists.org/content/early/2020/02/04/jeb.217174.abstract


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## BernhardHeuvel

Chronic exposure to glyphosate induces transcriptional changes in honey bee larva: A toxicogenomic study☆
Diego E.Vázquez, José M.Latorre-Estivalis, SheilaOnscWalter M.Farina
Environmental Pollution
Volume 261, June 2020, 114148
https://doi.org/10.1016/j.envpol.2020.114148

https://www.sciencedirect.com/science/article/pii/S0269749119367090


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## BernhardHeuvel

*Widespread contamination of wildflower and bee-collected pollen with complex mixtures of neonicotinoids and fungicides commonly applied to crops*

Environment International
Volume 88, March 2016, Pages 169-178
https://doi.org/10.1016/j.envint.2015.12.011
https://www.sciencedirect.com/science/article/pii/S0160412015301161

Highlights
• Pollen of oilseed rape and wildflowers growing nearby contained a wide range of pesticides.
• Pollen collected by honeybees and bumblebees also contained a broad range of pesticides.
• Pesticide exposure of bumblebee colonies in urban areas was lower than in rural areas.

Abstract
There is considerable and ongoing debate as to the harm inflicted on bees by exposure to agricultural pesticides. In part, the lack of consensus reflects a shortage of information on field-realistic levels of exposure. Here, we quantify concentrations of neonicotinoidinsecticides and fungicides in the pollen of oilseed rape, and in pollen of wildflowers growing near arable fields. We then compare this to concentrations of these pesticides found in pollen collected by honey bees and in pollen and adult bees sampled from bumble bee colonies placed on arable farms. We also compared this with levels found in bumble bee colonies placed in urban areas. Pollen of oilseed rape was heavily contaminated with a broad range of pesticides, as was the pollen of wildflowers growing nearby. Consequently, pollen collected by both bee species also contained a wide range of pesticides, notably including the fungicides carbendazim, boscalid, flusilazole, metconazole, tebuconazole and trifloxystrobin and the neonicotinoids thiamethoxam, thiacloprid and imidacloprid. In bumble bees, the fungicides carbendazim, boscalid, tebuconazole, flusilazole and metconazole were present at concentrations up to 73 nanogram/gram (ng/g). It is notable that pollen collected by bumble bees in rural areas contained high levels of the neonicotinoids thiamethoxam (mean 18 ng/g) and thiacloprid (mean 2.9 ng/g), along with a range of fungicides, some of which are known to act synergistically with neonicotinoids. Pesticide exposure of bumble bee colonies in urban areas was much lower than in rural areas. Understanding the effects of simultaneous exposure of bees to complex mixtures of pesticides remains a major challenge.


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## BernhardHeuvel

A bit older, but we missed it here.

*Intersections between neonicotinoid seed treatments and honey bees*

Christian H, Krupke Elizabeth, Y.Long
https://doi.org/10.1016/j.cois.2015.04.005
Current Opinion in Insect Science
Volume 10, August 2015, Pages 8-13

Synthesis and future directions
The additive effects of these various exposure routes are still being quantified. However, given the area devoted to production of crops grown from neonicotinoid-treated seeds, it is clear that a great degree of temporal and spatial overlap exists between neonicotinoids and pollinators and other non-target organisms. Exposure can take place through various matrices*—*including air-borne and stationary dusts, soil, plant products, and water. For honey bees, where most current research is focused, future estimates of individual and colony-level effects of these exposures should incorporate these multiple routes into assessments of risk posed by neonicotinoid residues. Of particular interest is the typical period of sowing of many annual crops grown from neonicotinoid-treated seeds, which corresponds closely with flowering of spring blossoms and the concomitant increase in honey bee foraging activity across the landscape [45].


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## BernhardHeuvel

*Thiamethoxam impairs honey bee visual learning, alters decision times, and T increases abnormal behaviors*

Learning is important for honey bee fitness and the pollination services that they provide. Neonicotinoid pesticides impair learning, fitness, colony health, and pollination, but most studies on how they affect bee learning have focused on olfactory learning. We tested the effects of field realistic doses of 0.8 ng/bee and 1.34 ng/bee of the neonicotinoid pesticide, thiamethoxam (TMX), on bee visual learning. We adapted a T-maze bioassay and classically conditioned bees to associate sugar reward with a simulated flower color (blue or yellow light) in a choice assay. At 1.34 ng/bee, TMX significantly reduced correct choices in the final learning trial as compared to the control treatment. There was no TMX effect in our 1-h memory test. We found stronger effects on decision time and abnormal behaviors. TMX decreased bee decision times, a potential byproduct of induced hyperactivity since bees walked to make choices. Behaviors (falling, trembling, and rapid abnormal movements) were sig- nificantly increased by both TMX doses as compared to the control treatment. These results suggest that the effects of neonicotinoids on bee visual learning should be further studied and incorporated into Risk Assessment protocols.

Ecotoxicology and Environmental Safety
Volume 193, 15 April 2020, 110367
Joshua C. Ludicke, James C.Nieh
https://doi.org/10.1016/j.ecoenv.2020.110367


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## BernhardHeuvel

*The miticide thymol in combination with trace levels of the neonicotinoid imidacloprid reduces visual learning performance in honey bees (Apis mellifera)
*

Abstract
Despite growing concerns over the impacts of agricultural pesticides on honey bee health, miticides (a group of pesticides used within hives to kill bee parasites) have received little attention. We know very little about how miticides might affect bee cognition, particularly in interaction with other known stressors, such as crop insecticides. Visual learning is essential for foraging bees to find their way to flowers, recognize them, and fly back to the nest. Using a standardized aversive visual conditioning assay, we tested how field exposure to three pesticides affects visual learning in European honey bees (Apis mellifera). Our pesticides were two common miticides, thymol in the commercial formulation Apiguard® and tau-fluvalinate in the formulation Apistan® and one neonicotinoid, imidacloprid. We found no effect of miticides alone, nor of field-relevant doses of imidacloprid alone, but bees exposed to both thymol and imidacloprid showed reduced performance in the visual learning assay.


Colin, T., Plath, J.A., Klein, S. et al. The miticide thymol in combination with trace levels of the neonicotinoid imidacloprid reduces visual learning performance in honey bees (Apis mellifera). Apidologie (2020). https://doi.org/10.1007/s13592-020-00737-6


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## BernhardHeuvel

*Effects of thiamethoxam and spinosad on the survival and hypopharyngeal glands of the African honey bee (Apis mellifera intermissa)*

Abstract
Insecticides can affect development and survival of non-target and beneficial arthropods like honey bees (Apis mellifera L.). Thiamethoxam and spinosad are widely used as pesticides in agriculture but they have become an important concern for beekeepers and researchers focusing on bee health; multiple reports stressed adverse effects on bees, notably on honey bees. The present study aims to evaluate the impact of these two insecticides on the development of the HPGs and on the survival of Apis mellifera intermissa a native African subspecies of honey bee present in Algeria. Newly emerged workers were acutely and chronically exposed to thiamethoxam and spinosad through sugar syrup and pollen pastry. The effects of these insecticides were assessed by measuring the size of HPGs acini and the total head protein content. The survival of the workers was also evaluated over 60 days when they were chronically exposed to the insecticides at concentrations corresponding to LC25 and LC10. We found that the insecticide-treated workers, after both acute and chronic exposure, exhibited smaller and irregularly shaped HPG acini. The total head protein content also decreased in treated individuals with the two concentrations of insecticides at day 6 and 9 compared to the respective controls. While the control group exhibited an LT50 (i.e. the time needed to kill 50% of the tested workers) of 22 days, the LT50 was only 3 days for the workers exposed to the LC25 of spinosad and all workers were dead at day 17. In contrast, thiamethoxam exposure at LC25 had no significant detrimental effect on honey bee survival. This study demonstrated the toxicity of thiamethoxam and spinosad to workers of A. mellifera intermissa and highlighted potential detrimental effects of the bioinsecticide spinosad on HPGs and survival of the bee workers.


Menail, Ahmed H.; Boutefnouchet-Bouchema, Wided F.; Haddad, Nizar; Taning, Clauvis N.T.; Smagghe, Guy; Loucif-Ayad, Wahida
Effects of thiamethoxam and spinosad on the survival and hypopharyngeal glands of the African honey bee (Apis mellifera intermissa)
Entomologia Generalis (2020)
DOI: 10.1127/entomologia/2020/0796


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## BernhardHeuvel

*Characterizing honey bee exposure and effects from pesticides for chemical prioritization and life cycle assessment*

Highlights
• Model for quantifying pesticide field exposure and ecotoxicity effects of bees.

• Integration of bee impacts in risk screening and life cycle impact assessment.

• Defined bee intake and dermal contact fractions as novel metrics for exposure.

• Case study on two pesticides on oilseed rape affecting 1260–1,360,000 bees/kg applied.

• Nectar foragers are the most affected forager type, with 32–190 ppm pesticide intake.


Abstract
Agricultural pesticides are key contributors to pollinator decline worldwide. However, methods for quantifying impacts associated with pollinator exposure to pesticides are currently missing in comparative risk screening, chemical substitution and prioritization, and life cycle impact assessment methods. To address this gap, we developed a method for quantifying pesticide field exposure and ecotoxicity effects of honey bees as most economically important pollinator species worldwide. We defined bee intake and dermal contact fractions representing respectively oral and dermal exposure per unit mass applied, and tested our model on two pesticides applied to oilseed rape. Our results show that exposure varies between types of forager bees, with highest dermal contact fraction of 59 ppm in nectar foragers for lambda-cyhalothrin (insecticide), and highest oral intake fractions of 32 and 190 ppm in nectar foragers for boscalid (fungicide) and lambda-cyhalothrin, respectively. Hive oral exposure is up to 115 times higher than forager oral exposure. Combining exposure with effect estimates yields impacts, which are three orders of magnitude higher for the insecticide. Overall, nectar foragers are the most affected forager type for both pesticides, dominated by oral exposure. Our framework constitutes an important step toward integrating pollinator impacts in chemical substitution and life cycle impact assessment, and should be expanded to cover all relevant pesticide-crop combinations.

Environment International
Volume 138, May 2020, 105642
Characterizing honey bee exposure and effects from pesticides for chemical prioritization and life cycle assessment
Eleonora Crenna, Olivier Jolliet, Elena Collina, Serenella Sala, Peter Fantke
https://doi.org/10.1016/j.envint.2020.105642


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## BernhardHeuvel

*A Review of Sub-lethal Neonicotinoid Insecticides Exposure and Effects on Pollinators*

Abstract
Purpose of Review
Beekeepers around the world have been reporting the ongoing weakening of honeybee health and subsequently the increasing colony losses since 1990. However, it was not until the abrupt emergence of colony collapse disorder (CCD) in the 2000s that has raised the concern of losing this important perennial pollinator. In this report, we provide a summary of the sub-lethal effects of pesticides, in particular of neonicotinoids, on pollinators’ health from papers published in peer-review journals.

Recent Findings
We have identified peer-review papers that are relevant to examine the effects of sub-lethal pesticide exposures on the health of honeybees (Apis mellifera), bumblebees (Bombus terrestris), and other bees from a literature search on PubMed and Google Scholar using the following combined keywords of “pollinators,” “honeybee,” “bees,” “pesticides,” or “neonicotinoids,” and from a cross-reference check of a report made available by the European Parliament in preparation to fulfill their regulatory mandate on the issue of protecting pollinators among their membership nations.

Summary
The weight-of-evidence of this review clearly demonstrated bees’ susceptibility to insecticides, in particular to neonicotinoids, and the synergistic effects to diseases that are commonly present in bee colonies. One important aspect of assessing and managing the risks posed by neonicotinoids to bees is the chronic effects induced by exposures at the sub-lethal levels. More than 90% of literature published after 2009 directly or indirectly demonstrated the adverse health effects associated with sub-lethal exposure to neonicotinoids, including abnormal foraging activities, impaired brood development, neurological or cognitive effects, and colony collapse disorder.

Lu, C., Hung, Y. & Cheng, Q. A Review of Sub-lethal Neonicotinoid Insecticides Exposure and Effects on Pollinators. Curr Pollution Rep (2020). https://doi.org/10.1007/s40726-020-00142-8
https://link.springer.com/article/10.1007/s40726-020-00142-8


References for further review/research:

1.
Abbott VA, Nadeau JL, Higo HA, Winston ML. Lethal and sublethal effects of imidacloprid on Osmia lignaria and clothianidin on Megachile rotundata (Hymenoptera: Megachilidae). J Econ Entomol. 2008;101(3):784–96.

2.
Alaux C, Brunet JL, Dussaubat C, Mondet F, Tchamitchan S, Cousin M, et al. Interactions between Nosema microspores and a neonicotinoid weaken honeybees (Apis mellifera). Environ Microbiol. 2010;12(3):774–82.

3.
Australian Government. Overview report neonicotinoids and the health of honey bees in Australia 2014. archive.apvma.gov.au/news_media/docs/neonicotinoids_overview_report_february_2014.pdf. Accessed 12 May 2018.

4.
Bernal J, Garrido-Bailón E, Del Nozal MJ, González-Porto AV, Martín-Hernández R, Diego JC, et al. Overview of pesticide residues in stored pollen and their potential effect on bee colony (Apis mellifera) losses in Spain. J Econ Entomol. 2010;103(6):1964–71.

5.
Biddinger DJ, Robertson JL, Mullin C, Frazier J, Ashcraft SA, Rajotte EG, et al. Comparative toxicities and synergism of apple orchard pesticides to Apis mellifera (L.) and Osmia cornifrons (Radoszkowski). PLoS One. 2013;8(9):e72587.

6.
Blanchard P, Schurr F, Celle O, Cougoule N, Drajnudel P, Thiéry R, et al. First detection of Israeli acute paralysis virus (IAPV) in France, a dicistrovirus affecting honeybees (Apis mellifera). J Invertebr Pathol. 2008;99:348–50.

7.
Boily M, Sarrasin B, Deblois C, Aras P, Chagnon M. Acetylcholinesterase in honey bees (Apis mellifera) exposed to neonicotinoids, atrazine and glyphosate: laboratory and field experiments. Environ Sci Pollut Res Int. 2013;20(8):5603–14.

8.
Bryden J, Gill RJ, Mitton RA, Raine NE, Jansen VA. Chronic sublethal stress causes bee colony failure. Ecol Lett. 2013;16:1463–9.

9.
Catae AF, Roat TC, De Oliveira RA, Ferreira Nocelli RC, Malaspina O. Cytotoxic effects of thiamethoxam in the midgut and Malpighian tubules of Africanized Apis mellifera (Hymenoptera: Apidae). Microsc Res Tech. 2014;77:274–81. https://doi.org/10.1002/jemt.22339.

10.
Cepero A, Ravoet J, Gómez-Moracho T, Bernal JL, Del Nozal MJ, Bartolomé C, et al. Holistic screening of collapsing honey bee colonies in Spain: a e study. BMC Res Notes. 2014;7(1):649.

11.
Chauzat MP, Faucon JP, Martel AC, Lachaize J, Cougoule N, Aubert M. A survey of pesticide residues in pollen loads collected by honey bees in France. J Econ Entomol. 2006;99(2):253–62.

12.
Christopher Cutler G, Scott-Dupree CD. A field study examining the effects of exposure to neonicotinoid seed-treated corn on commercial bumble bee colonies. Ecotoxicology. 2014;23(9):1755–63.

13.
Cox-Foster DL, Conlan S, Holmes EC, Palacios G, Evans JD, Moran NA, et al. Metagenomic survey of microbes in honey bee colony collapse disorder. Science. 2007;318:283–7.

14.
Cresswell JE, Page CJ, Uygun MB, Holmbergh M, Li Y, Wheeler JG, et al. Differential sensitivity of honey bees and bumble bees to a dietary insecticide (imidacloprid). Zoology (Jena). 2012;115(6):365–71. https://doi.org/10.1016/j.zool.2012.05.003.

15.
Cresswell JE, Robert FX, Florance H, Smirnoff N. Clearance of ingested neonicotinoid pesticide (imidacloprid) in honey bees (Apis mellifera) and bumblebees (Bombus terrestris). Pest Manag Sci. 2014;70(2):332–7.

16.
Cutler CG, Scott-Dupree CD, Drexler DM. Honey bees, neonicotinoids and bee incident reports: the Canadian situation. Pest Manag Sci. 2014;70(5):779–83.

17.
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## BernhardHeuvel

Honey bee (Apis mellifera) gut microbiota promotes host endogenous detoxification capability via regulation of P450 gene expression in the digestive tract

Summary
There is growing number of studies demonstrating a close relationship between insect gut microbiota and insecticide resistance. However, the contribution of the honey bee gut microbiota to host detoxification ability has yet to be investigated. In order to address this question, we compared the expression of cytochrome P450s (P450s) genes between gut microbiota deficient (GD) workers and conventional gut community (CV) workers and compared the mortality rates and the pesticide residue levels of GD and CV workers treated with thiacloprid or tau-fluvalinate. Our results showed that gut microbiota promotes the expression of P450 enzymes in the midgut, and the mortality rate and pesticide residue levels of GD workers are significantly higher than those of CV workers. Further comparisons between tetracycline-treated workers and untreated workers demonstrated that antibiotic-induced gut dysbiosis leads to attenuated expression of P450s in the midgut. The co-treatment of antibiotics and pesticides leads to reduced survival rate and a significantly higher amount of pesticide residues in honey bees. Taken together, our results demonstrated that honey bee gut symbiont could contribute to bee health through the modification of the host xenobiotics detoxification pathways and revealed a potential negative impact of antibiotics to honey bee detoxification ability and health.


Honey bee (Apis mellifera) gut microbiota promotes host endogenous detoxification capability via regulation of P450 gene expression in the digestive tract
Yuqi Wu, Yufei Zheng, Yanan Chen, Shuai Wang, Yanping Chen, Fuliang Hu, Huoqing Zheng
Microbial Biotechnology 2020 Apr 27. doi: 10.1111/1751-7915.13579
https://sfamjournals.onlinelibrary.wiley.com/doi/pdfdirect/10.1111/1751-7915.13579


Discussion
Cytochrome P450 enzymes are the major contributors to honey bee detoxification (Berenbaum and Johnson, 2015). In the current study, we focused on the interaction of honey bee gut microbiota and honey bee endogenous detoxification enzyme. Given the important role that honey bee P450 monooxygenase enzymes play in detoxification, our study provides important insights into the functional roles of gut bacteria as well as the interac- tions between gut microbiota and host detoxification capability in the honey bee.
The midgut is one of the main sites for detoxification in insects (Smagghe and Tirry, 2001). Pesticides can be taken up by the midgutepithelial cells, where most of it is metabolized before being transported back into the midgut lumen across the apical membrane or into the haemolymph across the basal membrane (Esther et al., 2017). Our results showed that gut microbiota strongly promotes the expression of key enzymes of the honey bee xenobiotic detoxification pathway. Six of the seven honey bee P450 detoxifying enzymes were upregulated in the midgut of CV workers, suggesting that honey bee gut microbiota enhance host detoxification capability and manipulate host metabolism. This is in accordance with related researches on mammals that demonstrated the importance of microbial activity in metabolic phenotype development. Toda et al. (Toda et al., 2009) reported that most of the major CYP isozymes were highly expressed in the livers of specific-pathogen-free mice compared with germ-free mice. Claus et al. (Claus et al., 2011) found that microbiota stimulates the expression and activity of major hepatic drug-metabolizing P450s. In the meanwhile, it is quite interesting to note that the P450 expressions in the hindgut were not influenced, though most of the honey bee core gut bacteria are colonized in the hindgut instead of the midgut (Martinson et al., 2012). The results that gut microbiota only influenced the P450 expression in honey bee midguts suggests that honey bee gut microbiota may have a different effect on the different parts of the gastrointestinal tract, which may be correlated with the bacterial abundance and composition differences (Martinson et al., 2012) or the physiological differences among different compartments of the bee gut. Therefore, further studies are needed to determine which cellular mechanisms underlie the observed regulatory function of gut microbiota and to explain why the P450 expressions are only influenced in the midgut. Collectively, our findings on the expression change of P450s indicated that gut microbiota has a strong positive effect on honey bee detoxification enzyme expression, which are vital to the detoxification ability and insecticide resistance of honey bees.
Neonicotinoid insecticides are an important group of neurotoxins specifically acting as antagonists of the insect nicotinic acetylcholine receptors (Matsuda et al., 2001). Currently, neonicotinoid insecticides are consid- ered as one of the main threats to honey bee health. Many lethal and sublethal effects of neonicotinoid insecticides on bees have been described in laboratory and field studies over the past decades (Blacquiere et al., 2012; Rundlo€f et al., 2015; Tsvetkov et al., 2017).

Of all the widely used neonicotinoid insecticides, thiacloprid has relatively low toxicity to honey bees (Iwasa et al., 2004), due to the fact that CYP9Q3 can metabolize thiacloprid with high efficiency (Manjon et al., 2018). Pyrethroids exert their toxic effects by disrupting the function of voltage-gated sodium channels which are critical for electrical signalling in the nervous system (Soderlund and Bloomquist, 1989). Tau-fluvalinate, a typical pyrethroid pesticide, is widely used in honey beehives as an acaricide for the control of devastating Varroa mites. The long-term application of fluvalinate as an apicultural tool as well as its absorption by the wax in the hive have resulted in a high-level of fluvalinate residue in bee colonies all over the world (Johnson et al., 2010). Fluvalinate is considered harmless to bees under normal circumstances (Johnson et al., 2006), because members of the honey bee CYP9Q subfamily, namely CYP9Q1, CYP9Q2 and CYP9Q3can efficiently metabolize fluvalinate (Mao et al., 2011). As expected, GDT bees administered with thiacloprid displayed a dramatically increased mortality rate and a higher level of thiacloprid residues compared with CVT workers. We also found that the innoxious fluvalinate became fatal when applied to GD workers, and the fluvalinate remaining in GDT workers was significantly higher than in CVT workers. 

Clearly, these findings showed that honey bee gut bacteria influence the metabolism of pesticide and confirmed that gut microbiota is crucial to honey bees for their pesticide tolerance. 

Still, there are plenty of studies that showed that insect bacteria have the ability to metabolize pesticides directly (Cheng et al., 2017; Dada et al., 2019), so we conducted an in vitro experiment to examine the possibility that the gut microbiota directly detoxifies the chemical pesticides and leads to resistance. Our results showed that neither of these two pesticides were significantly degraded by honey bee whole gut cultures in vitro, suggesting that the resident honey bee gut bacteria are not likely to degrade these two pesticides. However, future works using isolated bacteria strains are needed to provide a better understanding of the direct detoxification ability of honey bee gut symbionts. Then we have co-treated CV workers with both PBO and pesticide, we found that PBO treatment significantly reduced the honey bee survival rate, which is in accordance with previous studies (Iwasa et al., 2004; Johnson et al., 2006) and provided direct evidence for the involvement of P450 enzymes in thiacloprid and fluvalinate detoxification in the presence of the microbiota. Taken together, our results revealed that honey bee gut microbiota enhances host resistance to thiacloprid and fluvalinate through the regulation of the host endogenous detoxification mechanism, instead of direct degradation of toxins by gut symbiont. In addition, considering that P450 enzymes are capable of oxidizing many different substrates (Munro et al., 2013), we believe that the contribution of gut microbiota enhanced P450 expression to honey bee pesticide resistance is not limited to these two pesticides investigated in our study. 
Antibiotics have been a cornerstone of innovation in the fields of public health, agriculture and medicine. However, recent studies have shed new light on the collateral damage they impart on the indigenous host-associated communities (Modi et al., 2014). Zhan et al. (Zhan et al., 2018) revealed that the oral bioavailability of triazine herbicides was significantly increased in the rats treated with ampicillin or antibiotic ****tails, which is a consequence of the alteration of hepatic metabolic enzyme gene expression and intestinal absorption-related proteome. In apiculture, antibiotics are frequently used in bee colonies to prevent bacterial infection. Recent studies have demonstrated that antibiotic exposure can disrupt both the size and composition of the honey bee gut microbiome (Raymann et al., 2017; Raymann et al., 2018a), resulting in impaired metabolism, weakened immunity and decreased survivorship (Li et al., 2017; Raymann et al., 2017; Li et al., 2019). In our study, tetracycline, a commonly used antibiotic in bee keeping, was employed in field doses to workers. We observed a significantly decreased the community size after 5days after antibiotic treatment and a decrease in survival rate, similar to previous studies (Li et al., 2017; Raymann et al., 2017). These confirmed a successful establishment of a gut dysbiosis worker model and once again proved the detrimental effect of antibiotic on honey bee longevity.

In light of our findings above, we further evaluated whether the gut microbiota dysbiosis caused by antibiotic has a negative impact on honey bee detoxification ability, which might be a problem we will encounter in beekeeping. As predicted, our results displayed that gut microbiota dysbiosis downregulated the expression of P450s in the midgut, therefore, attenuating the honey bees’ detoxification ability. Interestingly, the expression changes of P450 in the midgut caused by the gut microbiota dysbiosis are quite different from gut microbiota deficiency, and the expression of two P450s functioning to metabolize phytochemicals (CYP6AS3 and CYP6AS4) was induced in the hindgut of AT workers. This may be due to the difference of metabolites in the gut of AT and of GD workers and suggests that honey bee gut microbiota deficiency and dysbiosis have different impacts on host physiology. The administration of both thiacloprid and fluvalinate on AT workers led to significantly increased mortality compared with that of pesticide treated NF workers. The pesticide remaining in the AT workers was significantly increased, which was probably caused by the downregulation of P450s in the midgut. These results demonstrated that the application of antibiotics interrupts the P450 expression in honey bee digestive tracts and enhances the pesticide risks for honey bees, even those of low toxicity to honey bees. The doses of pesticides we applied in this study were higher than actual field levels (38), suggesting that the combination of antibiotics and pesticides might not lead to an acute death of workers in the field colonies. Still, it is possible to hypothesize that gut dysbiosis could enhance the sublethal effects of pesticides, especially during the overwintering period when workers are exposed to antibiotics and pesticides (43) for a long period of time, and eventually lead to colony loss. However, our experiments were carried out using caged bees in a laboratory environment only, where workers have no route for acquisition of the gut microbiota and normally do not defecate. Thus, the combinatory effects of antibiotics and pesticides in field colonies remain to be determined. Moreover, it is worth studying the impact on honey bee detoxification of other chemicals (Kakumanu et al., 2016; Motta et al., 2018; Nogrado et al., 2019), that also perturb the gut microbial balance in honey bees.

Conclusion
Here in this study, our work revealed the interaction between honey bee gut microbiota and host resistance to pesticides for the first time. Our results showed that honey bee gut microbiota promotes the expression of detoxification enzymes in the midgut, which contribute to the host endogenous detoxification and resistance to thiacloprid and fluvalinate. These findings proved a close relationship between gut microbiota and honey bee detoxification capability, provided new insights into the honey bee host-microbiome interaction and perspectives for future studies on host-gut microbial metabolic interaction. In the current study, we have demonstrated a synergistic interaction between antibiotics and pesticides, which is detrimental for bees. And our results suggested this may be due to the reduced detoxification ability of honey bee. We were able to point out the beneficial role of a balanced gut microbiome in honey bees and provide fundamental information on how antibiotic treatment affects honey bee health.


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## BernhardHeuvel

Pesticide–Virus Interactions in Honey Bees: Challenges and Opportunities for Understanding Drivers of Bee Declines

Abstract

Honey bees are key agricultural pollinators, but beekeepers continually suffer high annual colony losses owing to a number of environmental stressors, including inadequate nutrition, pressures from parasites and pathogens, and exposure to a wide variety of pesticides. In this review, we examine how two such stressors, pesticides and viruses, may interact in additive or synergistic ways to affect honey bee health. Despite what appears to be a straightforward comparison, there is a dearth of studies examining this issue likely owing to the complexity of such interactions. Such complexities include the wide array of pesticide chemical classes with different modes of actions, the coupling of many bee viruses with ectoparasitic Varroa mites, and the intricate social structure of honey bee colonies. Together, these issues pose a challenge to researchers examining the effects pesticide-virus interactions at both the individual and colony level.

[...]

(B) Pesticide exposure can negatively impact many components and pathways of the immune system. Phagocytosis: pro-hemocyte differentiation can be impaired, resulting in fewer phagocytosing immune cells. The process of phagocytosis itself can also be affected; autophagy: Regulation of autophagy can be disrupted, potentially leading to apoptosis in cells; receptor activity: some insecticides target receptors that are also involved in antiviral defenses; gene expression: pesticides can alter expression of immune and detoxification genes. This includes upregulating inhibitors of the important immune system transcription factor, NF-κB; heat shock proteins: some pesticides downregulate expression of genes coding for heat shock proteins. These proteins can reduce viral load and also have functions in the RNAi antiviral pathway; gut bacteria: pesticides can also disrupt gut microbial communities, which are known to play roles in honey bee health and immunity.

Harwood, G.P.; Dolezal, A.G. Pesticide–Virus Interactions in Honey Bees: Challenges and Opportunities for Understanding Drivers of Bee Declines. Viruses 2020, 12, 566. https://doi.org/10.3390/v12050566


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## BernhardHeuvel

Effects of a Neonicotinoid Insecticide on the Growth of Honey Bee Gut Microbes

https://dc.ewu.edu/cgi/viewcontent.cgi?article=1028&context=srcw_2020_posters


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## BernhardHeuvel

Chronic within-hive video recordings detect altered nursing behaviour and retarded larval development of neonicotinoid treated honey bees

Abstract
Risk evaluations for agricultural chemicals are necessary to preserve healthy populations of honey bee colonies. Field studies on whole colonies are limited in behavioural research, while results from lab studies allow only restricted conclusions on whole colony impacts. Methods for automated long-term investigations of behaviours within comb cells, such as brood care, were hitherto missing. 

In the present study, we demonstrate an innovative video method that enables within-cell analysis in honey bee (Apis mellifera) observation hives to detect chronic sublethal neonicotinoid effects of clothianidin (1 and 10 ppb) and thiacloprid (200 ppb) on worker behaviour and development. 

In May and June, colonies which were fed 10 ppb clothianidin and 200 ppb thiacloprid in syrup over three weeks showed reduced feeding visits and duration throughout various larval development days (LDDs). On LDD 6 (capping day) total feeding duration did not differ between treatments. 

Behavioural adaptation was exhibited by nurses in the treatment groups in response to retarded larval development by increasing the overall feeding timespan. Using our machine learning algorithm, we demonstrate a novel method for detecting behaviours in an intact hive that can be applied in a versatile manner to conduct impact analyses of chemicals, pests and other stressors.

Siefert, P., Hota, R., Ramesh, V. et al. Chronic within-hive video recordings detect altered nursing behaviour and retarded larval development of neonicotinoid treated honey bees. Sci Rep 10, 8727 (2020). https://doi.org/10.1038/s41598-020-65425-y


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## BernhardHeuvel

Sublethal acetamiprid doses negatively affect the lifespans and foraging behaviors of honey bee (Apis mellifera L.) workers

Highlights
• Effects of sublethal acetamiprid doses on the foraging behaviors of Apis mellifera worker bees were firstly investigated using the RFID system under natural swarm conditions.

• Worker bees exposed to 2 μg/bee acetamiprid will induce precocious foraging activity and shortened the lifespan.

• Worker bees exposed to 2 μg/bee acetamiprid heavily decreased the workload throughout their lifetime.

• Excessive day-off rotation of worker bees exposed to 2 μg/bee acetamiprid is firstly reported.

Abstract
The neonicotinoid insecticide acetamiprid is applied widely for pest control in agriculture production. However, little is known about the effects of acetamiprid on the foraging behavior of nontarget pollinators. This study aims to investigate effects of sublethal acetamiprid doses on lifespans and foraging behaviors of honey bees (Apis mellifera L.) under natural swarm conditions. Newly emerged worker bees of each treatment received a drop of 1.5 μL acetamiprid solution (containing 0, 0.5, 1, and 2 μg/bee acetamiprid, diluted by water) on the thorax respectively. Bees from 2-day-old to deadline were monitored on foraging behaviors involving the age of bee for first foraging flights, rotating day-off status and the number of foraging flights using the radio frequency identification (RFID) system. We found that acetamiprid at 2 μg/bee significantly reduced the lifespan, induced precocious foraging activity, influenced the rotating day-off status and decreased foraging flights of worker bees. The abnormal behaviors of worker bees may be associated with a decline in lifespan. This work may provide a new perspective into the neonicotinoids that accelerate the colony failure.

Sublethal acetamiprid doses negatively affect the lifespans and foraging behaviors of honey bee (Apis mellifera L.) workers
Jingliang Shi et al., Science of The Total Environment, online 3 June 2020, 139924, https://doi.org/10.1016/j.scitotenv.2020.139924 
https://www.sciencedirect.com/science/article/pii/S0048969720334446


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## BernhardHeuvel

*Interaction of Varroa destructor and Sublethal Clothianidin Doses during the Larval Stage on Subsequent Adult Honey Bee (Apis mellifera L.) Health, Cellular Immunity, Deformed Wing Virus Levels and Differential Gene Expression*

Abstract: Honeybees (Apis mellifera L.) are exposed to many parasites, but little is known about interactions with abiotic stressors on their health, particularly when affected as larvae. Larvae were exposed singly and in combination to the parasitic mite Varroa destructor and three sublethal doses of the neonicotinoid insecticide clothianidin to evaluate their effects on survivorship, weight, haemocyte counts, deformed wing virus (DWV) levels and gene expression of the adult bees that subsequently developed. Clothianidin significantly reduced bee weight at the highest dose and was associated with an increase in haemocyte counts at the lowest dose, whereas V. destructor parasitism increased DWV levels, reduced bee emergence, lowered weight and reduced haemocyte counts. An interaction between the two stressors was observed for weight at emergence. Among the differentially expressed genes (DEGs), V. destructor infestation resulted in broader down-regulatory effects related to immunity that was often shared with the combined stressors, while clothianidin resulted in a broader up-regulatory effect more related to central metabolic pathways that was often shared with the combined stressors. Parasites and abiotic stressors can have complex interactions, including additive effects on reduced weight, number of up-regulated DEGs and biological pathways associated with metabolism.

[...]
However, the combination of V. destructor with clothianidin had additional effects observed on bee weight and the number of DEGs compared to the stressors alone. Thus, it appears that the combined stressors are able to have long-term effects on gene regulation, potentially affecting a broad range of biological pathways, which could affect the ability of the bees to metabolize and detoxify the neurotoxin, repair tissue damage or fight off infections like DWV that are essential for development and survival.

Morfin, N.; Goodwin, P.H.; Guzman-Novoa, E. Interaction of Varroa destructor and Sublethal Clothianidin Doses during the Larval Stage on Subsequent Adult Honey Bee (Apis mellifera L.) Health, Cellular Immunity, Deformed Wing Virus Levels and Differential Gene Expression. Microorganisms 2020, 8, 858. https://doi.org/10.3390/microorganisms8060858

https://www.mdpi.com/2076-2607/8/6/858


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## BernhardHeuvel

A documentation of neonics, bees and some chemical industry players in Europe. Translated into English. 

For your interest.

https://youtu.be/UaNSByf4sLA


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## BernhardHeuvel

Determination of Acute Lethal Doses of Acetamiprid and Cypermethrin for the Native Bee Apis mellifera (Hymenoptera: Apidae) in Cameroon

ABSTRACT
Honey bees are important pollinators and are essential in agriculture; as such they get exposed to a wide range of pesticides while foraging in contaminated fields or during the spray of chemical on crops. It is therefore important to know the toxicity and evaluate the impacts of bees’ exposure to these molecules. Acetamiprid and cypermethrin are two pesticides widely used in Cameroon and other countries. The objective of this study was to determine the toxicity of acetamiprid and cypermethrin on the native subspecies of Apis mellifera L. in agricultural areas in Adamaoua-Cameroon and to evaluate the impact on honeybee foragers exposed to lethal and sublethal doses of these two insecticides. The results obtained in laboratory conditions show that acetamiprid and cypermethrin are toxic to A. mellifera. The symptoms of neurotoxicity and first mortality appear 15 min after the ingestion of the high concentrations and about 30 to 45 min after the inoculation of the pesticides through contact route and the mortality increases with the concentration and time. The LC50 of acetamiprid obtained after 24 h are respectively 5.26 ng/μl for the topical application and 4.70 μg/μl by the oral route. At the same time, the LC50 of cypermethrin are respectively 2.27 ng/μl for topical application and 2.68 ng/μl for oral toxicity. For a sustainable agriculture and beekeeping, it is, therefore, important to establish quality measures on these insecticides in the ecosystem and to set up a phyto-pharmacovigilance and awareness system to the population.

Mazi, S., Vroumsia, T., Yahangar, M.-N., Malla, M. and Zroumba, D. (2020) Determination of Acute Lethal Doses of Acetamiprid and Cypermethrin for the Native Bee Apis mellifera (Hymenoptera: Apidae) in Cameroon. Open Journal of Ecology, 10, 404-417. https://doi.org/10.4236/oje.2020.107026


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## BernhardHeuvel

Long-term effects of neonicotinoid insecticides on ants

Abstract
The widespread prophylactic usage of neonicotinoid insecticides has a clear impact on non-target organisms. However, the possible effects of long-term exposure on soil-dwelling organisms are still poorly understood especially for social insects with long-living queens. Here, we show that effects of chronic exposure to the neonicotinoid thiamethoxam on black garden ant colonies, Lasius niger, become visible before the second overwintering. Queens and workers differed in the residue-ratio of thiamethoxam to its metabolite clothianidin, suggesting that queens may have a superior detoxification system. Even though thiamethoxam did not affect queen mortality, neonicotinoid-exposed colonies showed a reduced number of workers and larvae indicating a trade-off between detoxification and fertility. Since colony size is a key for fitness, our data suggest long-term impacts of neonicotinoids on these organisms. This should be accounted for in future environmental and ecological risk assessments of neonicotinoid applications to prevent irreparable damages to ecosystems.

Schläppi, D., Kettler, N., Straub, L. et al. Long-term effects of neonicotinoid insecticides on ants. Commun Biol 3, 335 (2020). https://doi.org/10.1038/s42003-020-1066-2

https://www.nature.com/articles/s42003-020-1066-2


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## BernhardHeuvel

Seek and you shall find: An assessment of the influence of the analytical methodologies on pesticide occurrences in honey bee-collected pollen with a systematic review

Highlights
• The analytical methodologies may underestimate the pesticide exposure on bees.
• Pesticide occurrence in pollen is negatively associated with the detection limit.
• In 4 pesticides, the detection limits were higher than their toxic doses.


Abstract
Honey bee mortality and colony losses have been reported worldwide. Although this phenomenon is caused by a combination of factors, agrochemicals have received special attention due to their potential effects on bees. In agricultural and urban environments bees are exposed to several compounds that may interact in unexpected ways, but information on the extent of pesticide exposure remains unclear. Several monitoring studies have been conducted to evaluate the field-realistic exposure of bees to pesticides after their release on the market. However, their outputs are difficult to compare and harmonize due to differences in the analytical methodologies and the sampling protocols (e.g. number of screened compounds and analysed samples, and detection limits (LODs)). Here, we hypothesize that the analytical methodologies used in the monitoring studies may strongly affect the pesticide occurrences in pollen underestimating the real pesticide exposure. By mean of a systematic literature review, we have collected relevant information on pesticide contaminations in the honey bee-collected pollen. Our findings showed that the pesticide occurrences were associated with the analytical methodologies and the real pesticide exposure has likely been underestimated in some monitoring studies. For four highly toxic compounds, the LOD used in these monitoring studies exceeded the doses that cause toxic effects on honey bees. We recommend that, especially for the highly toxic compounds, the LODs used in the monitoring studies should be low enough to exclude lethal or sublethal effects on bees and avoid “false negative” samples.


Gioele Toselli, Fabio Sgolastra; Seek and you shall find: An assessment of the influence of the analytical methodologies on pesticide occurrences in honey bee-collected pollen with a systematic review; Chemosphere; Volume 258, November 2020, 127358; https://doi.org/10.1016/j.chemosphere.2020.127358; https://www.sciencedirect.com/science/article/abs/pii/S0045653520315514


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## BernhardHeuvel

Insecticides cause transcriptional alterations of endocrine related genes in the brain of honey bee foragers

Highlights
• Endocrine activities of pesticides can be assessed in honey bee foragers.

• Pesticides caused transcriptional alterations in brain and HPG indicative of endocrine activity.

• Chlorpyrifos showed strongest transcriptional alterations.

• Affected genes encode proteins involved in transition of nurse and forager bees.

• buffy and mrjp down-regulation and hbg3 and ilp1 up-regulation are potential endocrine indicators.

Abstract
Bees are exposed to endocrine active insecticides. Here we assessed expressional alteration of marker genes indicative of endocrine effects in the brain of honey bees. We exposed foragers to chlorpyrifos, cypermethrin and thiacloprid and assessed the expression of genes after exposure for 24 h, 48 h and 72 h. Chlorpyrifos caused the strongest expressional changes at 24 h characterized by induction of vitellogenin, major royal jelly protein (mrjp) 2 and 3, insulin-like peptide (ilp1), alpha-glucosidase (hbg3) and sima, and down-regulation of buffy. Cypermethrin caused minor induction of mrjp1, mrjp2, mmp1 and ilp1. The sima transcript showed down-regulation at 48 h and up-regulation at 72 h. Exposure to thiacloprid caused down-regulation of vitellogenin, mrjp1 and sima at 24 h, and hbg3 at 72 h, as well as induction of ilp1 at 48 h. The buffy transcript was down-regulated at 24 h and upregulated at 48 h. Despite compound-specific expression patterns, each insecticide altered the expression of some of the suggested endocrine system related genes. 

Our study suggests that expressional changes of genes prominently expressed in nurse or forager bees, including down-regulation of buffy and mrjps and upregulation of hbg3 and ilp1 may serve as indicators for endocrine activity of insecticides in foragers.

Discussion
[...] 
However, the importance of these findings lies in the fact that there is an expressional change of these gene transcripts per se. The here studied insecticides chlorpyrifos, cypermethrin and thiacloprid induce neurotoxicity by different modes of action. This makes it plausible that the insecticides also differ to some extent in their transcriptional responses. Changes in expression of these hormone-associated genes in the worker bee brain and HPGs may translate to proteins, and subsequently, to physiological outcomes such as behavioral alterations of foragers.


Karl Fenta, Tiffany Haltiner, Petra Kunz, Verena Christen; Insecticides cause transcriptional alterations of endocrine related genes in the brain of honey bee foragers; Chemosphere
Available online 6 July 2020, 127542 In Press, Journal Pre-proof; https://doi.org/10.1016/j.chemosphere.2020.127542

https://www.sciencedirect.com/science/article/pii/S0045653520317379


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## BernhardHeuvel

A network analysis of how neonicotinoids have become embedded in New Zealand’s agricultural practices

7.1 Summary of research findings
The overall aim of this thesis was to understand how neonicotinoids have become embedded -and remain- in NZ’s agricultural practices. Subsequently, four research questions were identified and developed to reach this primary goal. The main findings of this research are presented below and are organised according to those research questions.
My first task was to identify and describe the major actors influencing neonicotinoid use in NZ. This began with an extensive review of publicly available documents which identified actors concerned with the use of neonicotinoids in NZ. The CSSF (Latour, 1999a) also help direct me towards those actors involved in the making and maintenance of agricultural practices. Once I had performed the initial interviews, I also used snowball sampling to try and encourage each participant to direct me towards individuals whom they thought would suitable for this research.
Some of the actors which this process identified were endemic insect pests whose abundance, and affinity to modified pastoral landscapes, have contributed to NZ’s reliance on agricultural chemicals like neonicotinoids and are used by other actors to convince one another of the importance of neonicotinoids. Next, there are landowners, they are the ones using seed treatment neonicotinoids on their properties to combat the vagaries of growing outdoors, meet the expectations of their consumers and maximise their yields. There are also beekeepers, who endure the adverse effects of neonicotinoids so that they can maintain relationships with landowners and secure access to valuable bee sites. Therefore, they are unlikely to complain if their hives are poisoned. Next, there are seed and chemical company representatives and product group representatives, these actors form strong relationships with international markets to secure market access for landowners, while simultaneously providing landowners with crop protection programmes, which include neonicotinoids, to ensure that a crop is produced. They also try and convince regulators not to ban neonicotinoids by demonstrating to them that neonicotinoids are vital for the production of key export crops. There are also international and domestic markets, which demand neonicotinoids are used in the production of hybrid seed crops and which self-regulate chemical residues on food. Lastly, there are the consumers, these actors demand that high yielding crops and food, which are free of pests, weeds, diseases and in some cases chemical residues, are available all year round.
My next task was to describe how certain relationships were formed and maintained between those actors. The interview questions attempted to draw out empirical evidence regarding
103
neonicotinoids and encouraged the interview participants to discuss their relationships with the other actors listed above. This identified several relationships centred on the use of neonicotinoids in NZ’s agricultural practices. To illustrate, analyse and discuss these relationships, Latour’s CSSF (1999) was once again applied to the research. In summary, I found that neonicotinoids are embedded -and remain- in NZ’s arable sector because of a series of material relations between arable growers, NZ seed and chemical companies, international seed companies, beekeepers, the NZEPA and consumers.
Firstly, what I have called the “arable profession” (autonomization) is formed by a three-way relationship between international seed companies, NZ seed and chemical companies and arable growers. International seed companies who demand high yielding and 100% seed crops interest NZ seed and chemical companies with payment and continued contracts to grow hybrid seeds in NZ, in return the NZ seed and chemical companies guarantee the production of 100% and high yielding seed crops by offering NZ’s more relaxed rules when it comes to some substances deemed hazardous elsewhere (neonicotinoids). The NZ based seed and chemical companies subsequently mobilise crop protection programmes, payment and access to markets to influence the farming practices of NZ’s arable growers. To secure their position NZ arable growers then mobilise a commitment to follow those crop management plans and use programs such as Agworld to confirm they have performed crop treatments as directed. Another relationship exists between arable growers and beekeepers (alliance). Essentially landowners convince beekeepers of their agricultural practices (use of neonicotinoids) by mobilising formal contracts, access to stances of highly valued Manuka and a fee for pollination services. Beekeepers similarly pay for access to honey-producing flora to interest arable growers in this relationship. Subsequently, due to the growth of NZ’s honeybee population and the Manuka honey market, those beekeepers with access to Manuka are unlikely to complain even in cases where their hives have been poisoned, this is due to the risk it poses their income. Next is the relationship is between the NZEPA, arable product groups and arable growers (alliance). Once again, this relationship relies on the trade of resources; product groups collect levies from growers based on yield, land area or at a fixed rate and in return and growers receive marketing, research and development as well as some practical support in return. Arable product groups also provide lobbying of the NZEPA, convincing them that neonicotinoids are essential for the success of key export crops, by mobilising critical use cases. Subsequently, the NZEPA is convinced of neonicotinoids importance to NZ’s agricultural production and are also convinced that biodiversity declines are not caused by neonicotinoids. In return the NZEPA do not changed neonicotinoid regulations. Finally, there is a relationship between the users of NZ grown hybrid (consumers) seeds and international seed companies (public representation). The international consumers interest the international seed companies with payment and those companies provide the consumers with the seed.
104

This demonstrates that each of the actors had a different role in the embedding and maintenance of neonicotinoids in NZ’s arable sector. Furthermore, the relationships between the actors are caused by their shared need to achieve their goals. Therefore, the actors are connected in a way that makes neonicotinoids indispensable for each of them to achieve their goals. It is in this regard that the actors make up the network, and their material relations serve to make and maintain neonicotinoids in NZ’s agricultural practices.
My third question was to explain why knowledge opposing the use of neonicotinoids has been unsuccessful in removing it from NZ’s agricultural practices. Like the above question, this was answered through the application of Latour’s CSSF (1999a) in Chapter 6, but its answer began during the data collection process. While performing the interviews, I found that the context surrounding an agricultural practice was present in the material relations I traced and therefore significantly impacted the application of neonicotinoids. Essentially, the embedding of an agricultural practice cannot simply be explained by the scientific resources supporting or opposing its use. To this end, I found that even the commodity being produced, and the agricultural sector was affecting practices. It became apparent very quickly that neonicotinoids were embedded in NZ’s arable sector but have been removed from NZ’s apples and pears sector. Through the application of Latour’s CSSF (1999a), I observed that a slight change in the “public representation” loop of the CSSF (Latour, 1999a, p. 103), circulated through all the other loops of the network and led to neonicotinoids being removed from the apples and pears sector’s pest control programmes. In summary, consumers of NZ’s apples and pears want to know what practices are contributing to the production of their food. Moreover, they want to make sure that chemicals which they are concerned about and are banned where they live (neonicotinoids) are not being used in the production of the food which they eat. Supermarkets who are directly connected to the consumer, through the transfer of resources, translate this demand into MRLs. Subsequently, to ensure MRLs are met and to maintain market access, apples and pears NZ have removed neonicotinoids from the crop protection plans, introducing an alternative and advising growers about changes in supermarket regulation and consumer demand. Apple and pear growers have therefore “tended to move away from them [neonicotinoids] and look for alternatives” (Product Group Representative 1). Thus, neonicotinoids have been removed from the agricultural practices of NZ’s apples and pears sector. This subsequently contradicts my initial assumption that neonicotinoids are embedded -and remain- in NZ’s agricultural practices.
Finally, I was charged with understanding how Latour’s CSSF (1999) could be used to investigate similar controversies in the future, in particular, those which involve hazardous substance use in NZ’s agriculture. To achieve this, in light of my research I critiqued and responded to critiques of Latour’s CSSF (1999a) and Callon’s key principles (1986). Firstly the principle “generalized symmetry” was vital for this research, by allowing me to highlight that MRLs are first shaped by international
105

supermarkets and then shape the crop protection programs produced by apples and pears NZ and therefore fresh product growers ending neonicotinoid use. Similarly, it enabled me to demonstrate the importance of grass grub in helping shape the crop protection programmes of NZ seed and chemical companies and therefore arable growers’ use of neonicotinoids. Secondly, by presenting the conflicting viewpoints of all the actors and quoting the exact words of the interview respondents, limiting my biased commentary, I further addressed critiques of “generalized symmetry” as well of those of “agnosticism” (see section 3.5). Essentially through the use of a thematic analysis I have still identified power relations and social hierarchies as they have emerged from the data. Next I addressed critiques of the CSSF’s imposition of its “own theoretical lexicon” (Whittle and Spicer, 2008). Essentially the CSSF’s interpretation by no means matches the world of the interview participants. This is largely because the vocabulary being used is vastly different from that used by the interview participants. To address this, I provided a thorough description of the case and the participants and presented many quotations which give insight into my translation of the interview participants’ responses. I also made a conscious effort to avoid using vocabulary from Callon (1986) and Latour (1999a) in the interview questions, so to avoid imposing this same vocabulary on the participants. Most importantly, Chapter 6 offers some insight into the major limitations and boundaries of myself and the research. By reflecting on my own limitations and the limitations of the tools which I used, I hope that future hazardous substance researchers will do a better job. To fully answer this question, the next section comments on what this research has contributed to CSSF and agricultural practice literature.

https://researcharchive.lincoln.ac.nz/bitstream/handle/10182/12152/Scott_Masters.pdf?sequence=4


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## BernhardHeuvel

Silicone Wristbands as Passive Samplers in Honey Bee Hives

Abstract: The recent decline of European honey bees (Apis mellifera) has prompted a surge in research into their chemical environment, including chemicals produced by bees, as well as chemicals produced by plants and derived from human activity that bees also interact with. This study sought to develop a novel approach to passively sampling honey bee hives using silicone wristbands. Wristbands placed in hives for 24 h captured various compounds, including long-chain hydrocarbons, fatty acids, fatty alcohols, sugars, and sterols with wide ranging octanol–water partition coefficients (Kow) that varied by up to 19 orders of magnitude. Most of the compounds identified from the wristbands are known to be produced by bees or plants. This study indicates that silicone wristbands provide a simple, affordable, and passive method for sampling the chemical environment of honey bees.

5. Conclusions
Considering the ease with which chemicals in bands can be compared, the minimal disturbance to the hive, and the variety of compounds detectable, using silicone bands to investigate the relationship between chemical compounds and honey bees shows great potential. Further, our results show that bands did not collect detectable compounds from outside of the hive, as no compounds were detected on the outside bands. This contrasts with SPME fibers, which are easily contaminated by background volatiles [16] and are quite expensive. Researchers can use bands as samplers in the open hive environment, as was done in this study, as well as in closed sampling containers. In a closed system, it would be possible to sample the volatile chemicals released by bees or adhered to the surface of bees based on certain castes, age groups, or environmental conditions, without the need for complicated air flow systems or filters. As a result, we believe using silicone band passive samplers provides alternative, flexible, more affordable opportunities to explore the chemical ecology of honey bees and the factors that influence their health, behavior, and survival.

Bullock, Emma & Schafsnitz, Alexis & Wang, Chloe & Broadrup, Robert & Macherone, Anthony & Mayack, Christopher & White, Helen. (2020). Silicone Wristbands as Passive Samplers in Honey Bee Hives. Veterinary Sciences. 7. 86. 10.3390/vetsci7030086.
https://www.researchgate.net/public...tbands_as_Passive_Samplers_in_Honey_Bee_Hives


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## BernhardHeuvel

Nature and Nurture: Effects of Multi-Pesticide Exposure during Honey Bee (Apis mellifera) Development

Honey bees (Apis mellifera) are frequently used as pollinators in a variety of agricultural production systems. While collecting resources, honey bee foragers can fly for miles around a colony, unintentionally gathering pesticides and sequestering synthetic chemicals inside a hive. Additionally, beekeeper-applied chemicals are a dominant component of most within-hive chemical environments. A diversity of pesticides residues can therefore be retained within hive components such as beeswax and stored pollen. These matrices can ultimately pose an exposure risk to developing brood. We used previously reported in-hive pesticide residues to test how mixtures of frequently encountered pesticides can impact honey bee development: 

1. We assessed the latent impacts of pesticides on adult queens by measuring how developmental exposure can impact a queen’s mating quality and the colony she later establishes, 

2. We examined how colony-level pesticide exposure influences the nutritional quality of diet fed by nurse bees to developing queen larvae, and 

3. We examined the differences in pesticide susceptibility and enzymatic detoxification across honey bee breeding stocks.

To investigate the effects of contact and oral multi-pesticide exposure during queen development, we reared queens in beeswax cups with or without an added multi-pesticide treatment and within colonies supplemented with treated (field-relevant pesticide mixture) or untreated pollen. We sacrificed queens post-mating to assess reproductive phenotype and established all remaining queens in standard hive equipment to measure colony growth. The colonies administered treated pollen produced fewer viable queens, and those queens which survived had significantly reduced stored sperm viability and tended to have a lower mating number. Furthermore, these queens later established colonies with lower brood viability. Our wax treatment contained a pesticide hazard-level similar to that of an average commercial colony and had no measureable effects. These findings indicate the downstream legacy effects of developmental exposure on queen mating quality and colony phenotype.

During development, honey bee queens are fed royal jelly (RJ) by nurse bees. We examined how pesticide exposure can influence the quality and quantity of RJ produced by a colony by exposing colonies to a multi-pesticide treatment in pollen and then harvested RJ from control and treated colonies. Thereafter, we measured the amount of RJ produced by colony, and screened samples for pesticide residues and nutritional composition. 

Colonies exposed to treated pollen yielded a lower mean amount of RJ provisioned per queen, but this difference was not significant. RJ from treated colonies contained lower amounts of phytosterols as well as key proteins and lipids relative to RJ from control colonies. We report that RJ from treated colonies has similar pesticide residues relative to controls, indicating that nurse bees buffer developing larvae from direct oral exposure to pesticides. This suggests that the effects of colony-level pesticide exposure on queen quality manifest through nutritional perturbations in RJ composition.

Susceptibility is a key aspect of pesticide risk which we compared across seven breeding stocks by rearing larvae in vitro using diet spiked with a pesticide mixture at four doses. We then tested differences in the activity of the detoxification enzyme esterase and used proteomics by mass spectrometry to investigate differential protein expression. 

We found that esterase activity towards two model substrates positively correlated with pesticide tolerance and the highly selected Pol-Line had larvae with the lowest pesticide tolerance and generally lower esterase activity. Conversely, larvae from progenitor and putatively feral stocks had the highest pesticide tolerance and esterase activity. 

We found few differences in the larval proteome across stocks, which indicates that differences in pesticide tolerance may result from qualitative differences in detoxification enzyme structure. This finding highlights the potential for unintended consequences of artificial selection on pesticide tolerance in honey bees.


MILONE, JOSEPH PERRY. Nature and Nurture: Effects of Multi-Pesticide Exposure during Honey Bee (Apis mellifera) Development (Under the direction of Dr. David Tarpy).

https://repository.lib.ncsu.edu/bitstream/handle/1840.20/38086/etd.pdf?sequence=1
https://repository.lib.ncsu.edu/handle/1840.20/38086


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## BernhardHeuvel

*Dietary quercetin impacts the concentration of pesticides in honey bees*

Highlights

• Honey bees orally exposed to imidacloprid exhibit reduced metabolism of quercetin.
• Dietary quercetin reduces the concentration of imidacloprid absorbed in honey bees.
• Dietary quercetin does not affect the concentration of tebuconazole or tau-fluvalinate in contact-exposed honey bees.

Abstract
Honey bees are important pollinators and are subject to numerous stressors, such as changing floral resources, parasites, and agrochemical exposure. Pesticide exposure has been linked to the decline in the global honey bee population. We have limited knowledge of the metabolic pathways and synergistic effects of xenobiotics in bees. Quercetin is one of the most abundant phytochemicals in plants and is therefore abundant in the honey bee diet. Quercetin can upregulate the detoxification system in honey bees; however, it is still unknown to what extent quercetin ingestion can reduce the content of absorbed pesticides. In this study, we investigated the effect of dietary quercetin on the contents of three pesticides in honey bees: imidacloprid (insecticide), tebuconazole (fungicide), and tau-fluvalinate (insecticide and acaricide). Bees were divided into two main groups and fed either quercetin-sucrose paste or only sucrose for 72 h. Thereafter, they were orally exposed to ∼10 ng/bee imidacloprid or contact-exposed to ∼0.9 μg/bee tau-fluvalinate or ∼5.2 μg/bee tebuconazole. After 1 h of oral exposure or 24 h of contact exposure, the bees were anaesthetised with CO2, sacrificed by freezing, and extracted with a validated QuEChERS method. Subsequently, the concentrations of the three pesticides and quercetin in the bees were determined with a triple quadrupole tandem mass spectrometer coupled to an HPLC system. No significant effect on the concentration of tebuconazole or tau-fluvalinate was observed in bees fed quercetin. Intake of quercetin led to a reduction in the concentration of imidacloprid in honey bees. Quercetin-rich plants may be exploited in future beekeeping.

Hamidreza Ardalani, Nanna Hjort Vidkjær, Bente B.Laursen et al., Dietary quercetin impacts the concentration of pesticides in honey bees, Chemosphere, Volume 262, January 2021, 127848, https://doi.org/10.1016/j.chemosphere.2020.127848

https://www.sciencedirect.com/science/article/pii/S0045653520320439

https://en.wikipedia.org/wiki/Quercetin


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## BernhardHeuvel

*Exposure to acetamiprid influences the development and survival ability of worker bees (Apis mellifera L.) from larvae to adults*

Highlights
• The response of immune- and detoxification-related genes in honeybees to acetamiprid decreased with exposure time.
• Acetamiprid disturbed the development of honeybees from larvae to adults.
• Continuously exposure to acetamiprid at 25 mg/L reduced the lifespan of adult worker bees.

Abstract
In most cases, honey bees experience pesticide pollution in a long-term period through direct or indirect exposure, such as the development process from larvae to the pre-harvest stage. At present, little is known about how honey bees respond to pesticide stresses during the continuous development period. This study aims to examine effects of long-term acetamiprid exposure on the development and survival of honey bees, and further present the expression profile in larvae, 1-day-old, and 7-day-old adult worker bees that related to immune, detoxification, acetylcholinesterase (AChE) and memory. Honey bees from 2-day-old larvae to 14-day-old adults except the pupal stage were continuously fed with different acetamiprid solutions (0, 5, and 25 mg/L). We found that acetamiprid over 5 mg/L disturbed the development involving birth weight and emergence rate of newly emerged bees, and reduced the proportion of capped cells of larvae at 25 mg/L; gene expression related to immune and detoxification of worker bees exposed to acetamiprid was roughly activated, returned and then inhibited from larval to emerged and to the late adult stage, respectively. Moreover, lifespans of bees treated with acetamiprid at 25 mg/L were significantly reduced. The present study reflects the potential risk for honey bees continuously exposed to acetamiprid in the development stage.

Jingliang Shi, Ruonan Zhang, Yalin Pei, Chunhua Liao, Xiaobo Wu, Exposure to acetamiprid influences the development and survival ability of worker bees (Apis mellifera L.) from larvae to adults, Environmental Pollution, Available online 12 August 2020, 115345, https://doi.org/10.1016/j.envpol.2020.115345
https://www.sciencedirect.com/science/article/abs/pii/S0269749120360334


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## BernhardHeuvel

*Toxicology of commercial insecticide formulations for the bees Scaptotrigona aff. depilis and Apis mellifera L.*

Résumé
Pesticides are believed to be responsible for the death of farmed and wild bees. Despite some studies on the active ingredients, little has been researched into the effects of commercial formulations, although there is evidence that inert materials potentiate the toxic effects of insecticides on bees. The present work aimed to assess the toxicity of three commercial products for bees
used in Brazil as a pesticide; Verismo® (Metaflumizon) and Actara® (Thiametoxam) in Apis mellifera and Evidence® (Imidacloprid) in Scaptotrigona aff. depilis should be examined comprehensively. The studies were carried out between 2016 and 2018 in the bee laboratory of the zootechnical department of the Federal University of Ceará and are based on the international protocols of the OECD (Organization for Economic Cooperation and Development).

Insecticide toxicity was assessed by different exposure routes using the topical average lethal dose (LD50) and the average lethal concentration per ingestion (LC50) for A. mellifera and the average lethal time (TL50), topically and orally for Scaptotrigona aff. depilis. Mortality data were recorded for Apis after 1, 6, 12, 24 and 48 hours and for 300 hours at intervals of 1, 6, 12 and 24 hours on the first day and every 24 hours from the second for Scaptotrigona due to insect exposure, with different Dosages of insecticides. For Apis the data were subjected to a log-logistic model or a Gompertz model with the help of the statistical environment R and for Scaptotrigona the data were subjected to a statistical analysis of the dose-response type using the Bioestat® program - version 5.3 . Actara® produced the DL50 topically 0.055 (24 h) and 0.052 (48 h) ng p.c./μL/abelha and the LC50 of uptake was 1.467 (24 h) and 0.095 (48 h) ng p.c./μL diet. Verismo® had a topical LD50 of 97.223 (24 h) and 1.157 (48 h) nL p.c./μL/abelha and the uptake LC50 was 2.493 (24 h) and 0.334 (48 h) nL p.c./μL diet. The topical TL50 Evidence® for Scaptotrigona aff. The depilis was 22.1 hours and 29.4 hours per intake.

The results showed that Actara is much more toxic than Verismo to A. mellifera regardless of the route of exposure. Evidence® is toxic to Scaptorigona aff. With depilis, however, the fatal effects of the product are more apparent when contamination occurs topically than when it is taken orally.

http://repositorio.ufc.br/bitstream/riufc/53300/3/2020_tese_lsgurgel.pdf


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## BernhardHeuvel

*Thiamethoxam exposure deregulates short ORF gene expression in the honey bee and compromises immune response to bacteria*

Abstract
Maximizing crop yields relies on the use of agrochemicals to control insect pests. One of the most widely used classes of insecticides are neonicotinoids that interfere with signalling of the neurotransmitter acetylcholine, but these can also disrupt crop-pollination services provided by bees. Here, we analysed whether chronic low dose long-term exposure to the neonicotinoid thiamethoxam alters gene expression and alternative splicing in brains of Africanized honey bees, Apis mellifera, as adaptation to altered neuronal signalling. We find differentially regulated genes that show concentration-dependent responses to thiamethoxam, but no changes in alternative splicing.

Most differentially expressed genes have no annotated function but encode short Open Reading Frames (sORFs), a characteristic feature of anti-microbial peptides. As this suggested that immune responses may be compromised by thiamethoxam exposure, we tested the impact of thiamethoxam on bee immunity by injecting bacteria. We show that intrinsically sub-lethal thiamethoxam exposure makes bees more vulnerable to normally non-pathogenic bacteria. Our findings imply a synergistic mechanism for the observed bee population declines that concern agriculturists, conservation ecologists and the public.

Thiamethoxam exposure deregulates short ORF gene expression in the honey bee and compromises immune response to bacteria
Pâmela Decio, Pinar Ustaoglu, Kamila Derecka, Ian C. W. Hardy, Thaisa C. Roat, Osmar Malaspina, Nigel Mongan, Reinhard Stöger, Matthias Soller
bioRxiv 853291; doi: https://doi.org/10.1101/853291 
https://www.biorxiv.org/content/10.1101/853291v3.full.pdf


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## BernhardHeuvel

*The effect of temperature on candidate gene expression in the brain of honey bee Apis*

*The effect of temperature on candidate gene expression in the brain of honey bee Apis mellifera (Hymenoptera: Apidae) workers exposed to neonicotinoid imidacloprid*

Highlights
• Extremely low imidacloprid neonicotinoid insecticide concentration in combination with contrasting temperatures causes gene expression changes associated to several honey bee (Apis mellifera) physiological responses.
• Greatly gene expression changes in hsp70, hsp90 and nalcn were triggered by effect of the low concentration dose in comparison with the higher concentration used.
• Cold conditions accentuate the alteration of transcription of nalcn gene in honey bees exposed to low doses of neonicotinoid.

Abstract
Neonicotinoid insecticides are potent agonists of nicotinic acetylcholine receptors and are a major factor in the decline of pollinators worldwide. Several studies show that low doses of this neurotoxin influence honey bee physiology, however, little is known about how insecticides interact with other environmental variables. We studied the effects of two neonicotinoid Imidacloprid doses (IMD, 0, 2.5, and 10 ppb), and three temperatures (20, 28, and 36°C) on gene expression in the brains of worker honey bees (Apis mellifera). Using qRT-PCR we quantified the expression of eight key genes related to the nervous system, stress response, and motor and olfactory capacities. Gene expression tended to increase with the low IMD dose, which was further intensified in individuals maintained in the cold treatment (20°C). At 20°C the octopamine receptor gene (oa1) was underexpressed in bees that were not exposed to IMD, but overexpressed in individuals exposed to 2.5 ppb IMD. Also, heat shock proteins (hsp70 and hsp90) increased their expression at high temperatures (36°C), but not with IMD doses. These results suggest that despite the low insecticide concentrations used in this study (a field-realistic dose), changes in gene expression associated with honey bee physiological responses could be induced. This study contributes to the understanding of how neonicotinoid residual doses may alter honey bee physiology.


Catalina Manzi, Jonathan Vergara-Amado, Lida Marcela Franco, Andrea X.Silva, The effect of temperature on candidate gene expression in the brain of honey bee Apis mellifera (Hymenoptera: Apidae) workers exposed to neonicotinoid imidacloprid, Journal of Thermal Biology, Available online 15 August 2020, 102696, https://doi.org/10.1016/j.jtherbio.2020.102696
https://www.sciencedirect.com/science/article/abs/pii/S030645652030468X


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## BernhardHeuvel

*Effects of selected insecticidal substances on mRNA transcriptome in larvae of Apis m*

*
Effects of selected insecticidal substances on mRNA transcriptome in larvae of Apis mellifera*

Highlights
• Loss of honeybee colonies in last decades.
• Insecticides are dealt as possible drivers.
• We fed different insecticides to honeybee larvae.
• Transcriptional analyses via RT-qPCR.
• Differences concerning development, energy metabolism and immune system.

Abstract
For the last decade, scientists have reported a loss of honeybee colonies. Multiple factors like parasites, pathogens and pesticides are dealt as possible drivers of honeybee losses. In particular, insecticides are considered as a major factor of pollinator poisoning. We applied sublethal concentrations of four insecticidal substances to honeybee larval food and analyzed the effects on transcriptome. The aim was to identify candidate genes indicating early negative impacts after application of insecticidal substances. Honeybee larvae were kept in-vitro under hive conditions (34–35 °C) and fed with dimethoate, fenoxycarb, chlorantraniliprole and flupyradifurone in sublethal concentrations between day 3–6 after grafting. Larvae at day 4, 6 and 8 were sampled and their transcriptome analyzed. By use of a RT-qPCR array differences in gene expression of selected gene families (immune system, development detoxification) were measured. Targets mainly involved in development, energy metabolism and the immune system were significantly affected by the insecticidal substances tested, selectively inducing genes of the detoxification system, immune response and nutritional stress.

Arne Kablau, Jakob H.Eckert, Jens Pistorius, SoroushSharbati, Ralf Einspanier, Effects of selected insecticidal substances on mRNA transcriptome in larvae of Apis mellifera, Pesticide Biochemistry and Physiology
Available online 10 September 2020, DOI: https://doi.org/10.1016/j.pestbp.2020.104703
https://www.sciencedirect.com/science/article/abs/pii/S004835752030198X


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## BernhardHeuvel

*Honey Bees and Neonicotinoid-Treated Corn Seed: Contamination, Exposure, and Effects.*

Abstract: Most corn (Zea mays) seeds planted in the United States in recent years are coated with a seed treatment containing neonicotinoid insecticides. Abrasion of the seed coating generates insecticide-laden planter dust that disperses through the landscape during corn planting and has resulted in many "bee-kill" incidents in North America and Europe. We investigated the linkage between corn planting and honey bee colony success in a region dominated by corn agriculture. Over 3 yr we consistently observed an increased presence of corn seed treatment insecticides in bee-collected pollen and elevated worker bee mortality during corn planting. Residues of seed treatment neonicotinoids, clothianidin and thiamethoxam, detected in pollen positively correlated with cornfield area surrounding the apiaries. Elevated worker mortality was also observed in experimental colonies fed field-collected pollen containing known concentrations of corn seed treatment insecticides. We monitored colony growth throughout the subsequent year in 2015 and found that colonies exposed to higher insecticide concentrations exhibited slower population growth during the month of corn planting but demonstrated more rapid growth in the month following, though this difference may be related to forage availability. Exposure to seed treatment neonicotinoids during corn planting has clear short-term detrimental effects on honey bee colonies and may affect the viability of beekeeping operations that are dependent on maximizing colony size in the springtime.

Environmental Toxicology and Chemistry, 05 Feb 2021, 40(4):1212-1221
DOI: 10.1002/etc.4957 PMID: 33289922 PMCID: PMC8048971






Europe PMC


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europepmc.org


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## BernhardHeuvel

*Acute oral exposure to imidacloprid induces apoptosis and autophagy in the midgut of honey bee Apis mellifera workers*

*Highlights*

•
Imidacloprid may affect non-target organs of honey _Apis mellifera_.
•
The ingestion of this chemical by bees causes ultrastructural changes in midgut.
•
Exposure to imidacloprid induces autophagy and apoptosis in the midgut of bees.

*Abstract*
The honey bee _Apis mellifera_ is an important pollinator that increases the yield and quality of crops. In recent years, honey bee populations have declined in some parts of the world, which has been associated with several causes, including pesticides used in agriculture. Neonicotinoids are neurotoxic insecticides widely used in the world with systemic action mode contaminating nectar and pollen that may be consumed by bees. This study evaluated the side effects of imidacloprid in the midgut of _A. mellifera_ after acute oral exposure. Toxicity, histopathology, cytotoxicity, and expression of autophagy-related gene _atg1_ were evaluated in honey bee workers orally exposed to imidacloprid. The estimated imidacloprid LC50 was 1.44 mg L−1. The midgut epithelium of bees fed on imidacloprid LC50 has the occurrence of cytoplasm vacuoles, enlarged intercellular spaces, disorganization of the striated border, and nuclear pyknosis, with an organ injury index that increases with time exposure. The midgut digestive cells of treated bees have apical protrusions, damaged mitochondria, and autophagosomes that were characterized for content with organelle debris and high expression of _atg1_. These features indicate the occurrence of high cell death in the midgut of workers exposed to imidacloprid, which may affect the digestibilitythe physiology of the insect.

Redirecting


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## Litsinger

BernhardHeuvel said:


> Exposure to seed treatment neonicotinoids during corn planting has clear short-term detrimental effects on honey bee colonies and may affect the viability of beekeeping operations that are dependent on maximizing colony size in the springtime.


Thanks for the link, Bernhard. Kirk Webster has intuitively come to this same conclusion:


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