# Nosema Sequential Sampling



## enjambres (Jun 30, 2013)

Ian,

Why that scope rather than the binocular Omana model recommended by Randy Oliver? Just for the digital capacity? (The binocular one is more expensive, though, IIRC @$347 USD.) I am trying to figure out which model to buy. It's a big ticket item for me so I am dithering.

Enj.


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## Ian (Jan 16, 2003)

cost yes, cool factor (yes yes yes, I know...) is the other. 
If it works as I have been advised, I think it will solve my non scientific nature of using microscopes
All I need to do is identify and count nosema and close up views of mites for biting damage, so I'm loving this idea 
:thumbsup:

I'll let you know if it lives up to my expectations


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## enjambres (Jun 30, 2013)

Well, I'd love to see chewed-upon mites, so be sure to post those pics - beekeepers' porn!

Enj.


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## Allen Martens (Jan 13, 2007)

Ian I think this type of sampling individual hives is much more useful than global sampling. That's the way I have done it the last couple of years.

Global sampling provides very little useful information IMHO other than possibly telling you that you have a monstrous problem and even that could be incorrect if you happened to sample several severely infected hives.


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## Ian (Jan 16, 2003)

I agree
My hold up til now was equipment.

What are you finding as for infection in your hives?


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## camero7 (Sep 21, 2009)

This is the one I have...I like it a lot and I can take it to the field and sample on the truck hood. Quick analysis


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## NY Bee Wellness (Dec 21, 2013)

Here is the link to the NY Bee Wellness video of Randy Oliver demonstrating two methods:

https://www.youtube.com/watch?v=6SFhDW_IxZ8


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## Ian (Jan 16, 2003)

Excellent video 
Thx!


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## Ian (Jan 16, 2003)

Has anyone done any work on nosema tolerant bees?


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## Allen Martens (Jan 13, 2007)

Ian said:


> I agree
> My hold up til now was equipment.
> 
> What are you finding as for infection in your hives?


I haven't sampled this year. Mostly, I sample for interest sake. I rarely do anything about nosema. 

I think late winter would be very useful in a queen breeding program. If it is possible to breed resistance to nosema, selecting hives with low infections rates from a wintering shed after 5 months of confinement is about as good a tool as you will find.

Interestingly, this time of year there are basically no slightly infected bees. They are either nosema free or blow your mind infected. I think this is why hives with higher infection rates do fine many times when they get moved out. The infect hives go for their first spring flight and probably never come back while the disease free bees keep going. I don't have proof for this but that is what seems to be happening to me.


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## Ian (Jan 16, 2003)

Ya good point
Late winter Sampling is the ideal time to test for nosema infection in regards to ranking for breeding efforts


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## Ro bert (Feb 26, 2017)

Hey Ian, you're mailbox is full! fyi.


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## nediver (May 26, 2013)

In watching the Randy Oliver Presentation posted by NY Bee Wellness it seems Randy doesn't treat Nosema. 

I read his article on the topic awhile back on how to sample, but why bother if your not going to treat. 

Do I have this wrong?


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## jean-marc (Jan 13, 2005)

I will agree with you Ian that the spore counting gives such inconsistent results. Numbers are all over the place, sometimes they don't even go down after using fumigillin. They generally go down after good cleansing flights but not always. They monitored pretty close for 3 years, spring and fall and concluded that I did not know any more than when I started. Numbers were all over the place all the tie with composite sampling. I never really tied it to survivabilty or to honey production. Counts were generally low, except when they weren't (lol). So we treat prophylactically. Perhaps it is time to re-examine the topic.

I sorta like your new tool. Keep up posted. I likely won't get one, it's just more work, and I'm not as curious as I used to be.

Jean-Marc


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## Ian (Jan 16, 2003)

I like this microscope but yes, binoculars would be better. The digital view is awesome but too cumbersome and not used during each quick slide assessment...so yes, the digital is just a toy 

I first counted a good looking hive...nothing, not a one. So then I tested from the hive in my topic photo upto, 8 out of 10 heavy heavy counts. 
Now to test a few random tests from my treatment trials


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## Ian (Jan 16, 2003)

nediver said:


> In watching the Randy Oliver Presentation posted by NY Bee Wellness it seems Randy doesn't treat Nosema.
> 
> I read his article on the topic awhile back on how to sample, but why bother if your not going to treat.
> 
> Do I have this wrong?


Why bother? >>To know<<

Here are my too common conversations with beekeepers of all stripes;

"... my hives crashed because of farmers..."
"...what's your disease levels and conditions?"
"...I don't know but that farmer sprays around here..."

We need to put more effort into knowing our hives conditions to be able to properly diagnose their troubles. EVERY other industry does it...beekeepers assume too much


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## Ian (Jan 16, 2003)

I just counted a slide with the nosema cheek buy cheek from one side of the shot to the other... yikes


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## Ian (Jan 16, 2003)

What are these egg yoke looking objects?


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## Michael Palmer (Dec 29, 2006)

I think pollen grains


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## Roland (Dec 14, 2008)

Ian, could you you include the power of magnification used for your photomicrographs? Or, do you have a calibrated grid you can superimpose? It might help in identification.

Crazy Roland


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## Ian (Jan 16, 2003)

Roland it's 400x, contents of my bees intestinal tract. 
Some bees are littered with these objects , others nothing. Randy mentions an egg yoke looking object to be rust yeast. Does this match rust yeast or as Michael suggested, simply pollen? 
This is my first time using a microscope since grade 11 Bio class... lol
A Facebook friend suggested using a drop of iodine and a thinner sample to help define the slide sample.


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## Ian (Jan 16, 2003)

One bee Chalk full,


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## Roland (Dec 14, 2008)

Ian, streaking was in style last time I looked thru a microscope. I can dig out my microbiology book tomorrow and try to jog some brain cells. Of the top of my head, various stains where use, such as Gram's and Picric acid. It helped determine what the blob might be. 

Do you see any surface irregularities? They looked smoth to me.

Crazy Roland


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## Ian (Jan 16, 2003)

I have a slide for you Roland , just give me a moment to load it. 
I've picked up a bunch of stuff chatting with guys about micro organisms. 
The pic earlier is half digested pollen, as Michael Palmer suggested. The pic I posted up top is one absolutely full of nosema spores. Typically don't see them that full, took a pic as my trophy lol


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## Ian (Jan 16, 2003)

The yellow yoke looking objects... I've gotten a few diagnosis ( hard diagnosing with unfocused pics on the Internet)
Either rust yeast, Nigel rein whatever that means or crithidia
All not good, but what can you do... looks cool
I just see these guys occasionally


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## jean-marc (Jan 13, 2005)

Crithidia are protozoans. They can be a problem in the wet areas like on the West Coast especially in the winter. They prevent bees from fully digesting the pollen they consume. This in turn does not allow bees to make good jelly for feeding larvae. Apparently the acidic content of honey prevents crithidia and other protozoans from reproducing and thriving. That is why in our case bees that goto Allberta for a honey flow do just fine in the winter. The ones that stay here don't always do great. They survive ok, but are not boiling at the seams like some of the Alberta bees do. All this informtion is verbatim from Caspian Apiaries.

Still waiting for signs of warmth and a descent pollen flow.

Jean-Marc


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## Roland (Dec 14, 2008)

Ian - how difficult would it be for you to obtain a small quantity if Gram's stain?

Crazy Roland


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## Maybee Apiaries (Jun 23, 2016)

Ian said:


> The background level of nosema in my hives is tremendous yet the visual symptoms of the infection has basically disappeared. Lab testing proves that almost the entire nosema infection is Ceranae.
> Existing Sampling and spore counting of various test groups shows such inconsistent results that I can't make sense of anything. So I'm officially scrapping composite Sampling and spore counts.
> I've been reading Randy Oliver's work on Sequential Sampling for some time now but I have not adopted his method due to time and equipment needed...until now. Hello compound microscope, hello new options.
> 
> ...


Hey Ian, where did you pickup this microscope? I looked on Amazon canada, but couldn't find any. Microscope.com has decent deals, but shipping from the states is $100+. 
Thanks in advance.


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## Ian (Jan 16, 2003)

Roland said:


> Ian - how difficult would it be for you to obtain a small quantity if Gram's stain?
> 
> Crazy Roland


My cousin is a vet so I'm sure he has stains or dyes on hand. I'll track some down. Do you have experience with staining samples? Because I don't have any.


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## Ian (Jan 16, 2003)

Maybee Apiaries said:


> Hey Ian, where did you pickup this microscope? I looked on Amazon canada, but couldn't find any. Microscope.com has decent deals, but shipping from the states is $100+.
> Thanks in advance.


MB, it cost me $90 shipping after exchange. 
The microscope I bought here is excellent, the digital is useful for a guy like me who likes to take pics and show anyone who listens,...lol
I think the binocular would be nice, but whatever, I don't plan on spending my life behind one.


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## Roland (Dec 14, 2008)

Ian asked"

Do you have experience with staining samples? Because I don't have any.


Oh sure, but that was back when Pintos, Vegas, and Beetles ruled the roads, and gas was expensive at 85 cents a gallon. I found my Microbiology book and realized I had forgotten more than I currently know. 

I will read more tomorrow and report back with suggestions. It was not hard, if you have all of the ingredients. 

What illumination does you microscope come with? Any polarizing filters or other gadgets? It may determine what route is best.

Crazy Roland


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## Ian (Jan 16, 2003)

Lol, ... I am good at picking out nosema spores... 
I wasn't sure what those polarizing filters did,


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## Ian (Jan 16, 2003)

I've been told that those yellow egg looking yoke objects look like oil or fat.


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## beebreeder (Nov 24, 2009)

Interesting thread this Ian, it confirms my suspicions that Beekeepers the world over do not do much microscope work and as you rightly say assume/blame outside influence on colony state of health, I thought it was just the UK that has this attitude


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## Roland (Dec 14, 2008)

There should be a selective stain for lipid(oils). I will look in my textbook tomorrow.

Crazy Roland


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## Ian (Jan 16, 2003)

>>>For what it’s worth; <<<

After running my nosema samples through Sequential Sampling to determine my apiarie’s 10% infection rate (safe zone), I gathered the remaining bees from each sample into a composite sample and sent them away for couunts at the NBDC. My counts have returned.
Of the 10% infected bees, my non treatment infected bees held a lower count (2m counts) as compared to my fumagillin treated infected bees (10m spore counts). Also my Thymol control group infected bees counted 7m spores. So to summarize from this, my apiary holds a low nosema infection rate (6-10% across the ALL test groups), but of those infected bees the non treatment group held lower actual counts than the treated groups. The low infection rate but higher counts per infected bee is why my samples come back so high yet my apiary seems to perform well. I need to watch my Infection rate, if it starts to climb up past 30% my apiary performance may suffer. But I don’t know what I would do about it. If my fumagillin trial tells me anything, treatment only makes Infection worst…

http://stepplerfarms.com/Honeyblog/?p=7826


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## nediver (May 26, 2013)

Ian said:


> Why bother? >>To know<<
> 
> Here are my too common conversations with beekeepers of all stripes;
> 
> ...


Ian thanks for your response and I agree on the being informed. Are you treating for Nosema? Certain threshold? Treatment of choice?


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## Ian (Jan 16, 2003)

All questions with no answers 
And IMO a huge issue which few take seriously


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## irwin harlton (Jan 7, 2005)

seems to be a case of , ****ed if you do and ****ed if you don't


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## Ian (Jan 16, 2003)

The problem is zero RnD into bee disease. In the livestock industry we have pretty much a treatment for every ailment. With bees we just shrug our shoulders and let them die


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## Roland (Dec 14, 2008)

Ian = how did you apply the Fumigillan? 


Crazy Roland


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## Ian (Jan 16, 2003)

Roland said:


> Ian = how did you apply the Fumigillan?
> 
> 
> Crazy Roland


stored syrup

Before you say degradation, let me make note that my nosema species has completely shifted over to Ceranae


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## Roland (Dec 14, 2008)

What would it take to do a comparison between syrup and "Red Ball of Fire"?

Crazy Roland


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## Ian (Jan 16, 2003)

I find Fumagillin is fumagillin regardless how it's administered. 
Are you finding efficacy in the RBF ?


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## Roland (Dec 14, 2008)

Only used RBoF once, saw DRAMATIC effect. 

Crazy Roland


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## Ian (Jan 16, 2003)

Why only once?


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## Roland (Dec 14, 2008)

Because it solved the problem.

Crazy Roland


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## James Kellie (Oct 30, 2010)

Novice at Nosema, could you tell me what the RBF treatment is and also I was told there is no treatment for Nosema Cerana that Fumigillan did not work for Cerana only Apis. Have I been miss informed.?


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## Roland (Dec 14, 2008)

Don't know which Nosema we had, but it killed 90 percent of the hives for 2 years. RBoF is a novel method of Fumigilan that we believe controls the release much better than in syrup. With syrup, some gets stored away and gets used at a possibly sub therapeutic level at a later time. We believe it is better to apply at a constant level, for slightly longer than it takes for the spores to pass thru the digestive tract. It does no good if the levels drop to below therapeutic before the digestive cycle is finished.

I could be mis-informed, but did not worry about it. We cured our problem.

Crazy Roland


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