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Discussion Starter #1
Million dollar question...

How do I accurately quantify nosema spores under a microscope?

I purchased a new microscope after reading Randy Oliver's article on Sick Bees – Part 13: Simple Microscopy of Nosema for Beekeepers.


BUT, I am still a bit confused on how to quantify nosema spores under a microscope.

Can anyone help me understand how to determine if we have too many Nosema spores?

Thank you,

Soar
 

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I'll take that million please... You should be able to find a slide with an etched gradient or pattern . I do not know if a defined test exist, but basically you would count the spores within a square.

Please pay at this window.

Crazy Roland
 

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It depends - sorry I could not resist 😈
First think about standard dilution - 1 milliliter of water per bee abdomen, usually a sample of 10 bees/10 ml. water well mashed and one drop on the slide with cover slip.
The area -size of what you see at 400 power is pretty much standard so count number of spores divide by 5.
5 spores in the field of view equates to 1,000,000 per bee.
Or you can buy one of the grid marked slides Roland mentioned to get more exact.
How many is a problem? Depends on who you ask 6-8 million?
Randy uses single bee squash and either they have spores or not looking for a % of infected bees per hive. Read the rest in his site.
Comparing the two methods the issue comes about that you could have one badly infected bee in a sample of ten and have an average of 6-8 million in 10.
 

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Discussion Starter #6
Soars with eagles - do you see Nosema symptoms?

Crazy Roland
Roland,

Thank you for your kindness in asking. This is a very painful story for me...

We started about 5 years ago after reading about bee colony collapse. I asked my wife if we should maybe start to keep honey bees [we have an orchard and live on a ranch]. We did a simple prayer: God, what are your thoughts on us getting honeybees? The very next day we came home and were shocked and amazed to find a monster swarm just a few feet from our front door. I began to do massive reading and asking questions. Within a year our numbers went vertical [we were catching 5-10 swarms a day during the peak of the swarm season]. For a two year period we were catching swarms faster than we could build the wood-ware. We also learned to make splits, graft queens, etc. So we began to experienced radical exponential growth. Two years ago I placed an ad on Craigslist to rent our honey bees to almond growers. I was thinking no one would call us. I was shocked to discover massive calls and people desperate for pollinators. So we began to take our honeybees 10 minutes away to the almond orchards...

Then the nightmare began. Last year, we went from 200 colonies all the way down to 4 colonies...so I think that is a 98% loss and I was crushed beyond words.

No one could give me a clear answer as to why. I have cared so much for the honeybees and most years we kept growing more and more...and it was one of the most wonderful endeavors we have ever done. It was very hard work [building boxes, lids, bottoms, bee stands, frames, etc.] and now I realize very expensive [I put my entire life savings into it].

Last winter, for the first time, we began to experience great losses. And I would be so happy to discover what it could be so I could rectify the problem. After losing 98% last winter I wanted to quit and get completely out of beekeeping because my dream was so utterly crushed I experienced a level of depression and emotional pain that I have never had. But after placing everything up for sale, it just did not feel right on the inside [I had no peace] so I only sold pallet fulls of unassembled boxes and frames and kept a few hundred boxes, lids and bottoms.

Here's some of the background:

We treat with oxilic acid using the ProVap 110 Sideliner/Commercial Oxalic Acid Vaporizer. To be honest with you, we only treat once or twice a year because for most of the year the Varroa mite load is nearly non existent. So I believe the problem cannot be the Varroa mite because we never permit the load levels to go high.

Two falls ago, for the first time ever, we began to see honey bees crawling all over on the ground within 100 feet of our bee yard. Last fall they did the same thing and they were dying.

The symptoms are the bee numbers begin to drop slowly in the late fall and continue to drop until the there is not one honeybee left in the box. The boxes are still filled with honey and pollen but not one darn honeybee inside! There is no outward sign of disease [example: no bad smell from EFB/AFB, no massive wax moth activity, no massive bee poop from Nosema apis, the honeybees simply begin to decline in numbers slowly but surely until not even one bee is left. We feed with sugar syrup and Mann Lake Ultra Bee Pollen.

This year, I stumbled upon Randy Oliver's description of Nosema ceranae. What stood out to me was that he said there were no outward signs of a problem...but there was a continual decline in bee population.

No one has been able to help me discover what is causing our honey bees to die in such massive numbers.

I thought I would buy a microscope and see if Nosema ceranae is the culprit.

I probably should not have done it, but I already began the treatment yesterday with the 12 strongest of our surviving colonies. This winter, we went from 126 colonies down to less than 50, so the losses are not as bad as last year, but there is still something terribly wrong because historically we have over wintered here with great success.

Ok, can someone please help us?

Thanks,

Soar
 

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Soar
Your story is touching and we feel your pain at losing colonies.
There is information lacking that may allow some of us to help.
You treat once or twice a year, what triggers that? When?
Are you giving a single dose of OA while broodless or a three dose 7 days apart?
How are you determining mite load which you said is not an issue.
You started treatment with Fumidil, did you buy the microscope and do the spore counts? What were they?
Your info about catching bees and expansion sounds like there are lots of other hives around so mites not being an issue may be an incorrect assumption.
Giving us data may help the analysis.
 

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look up BVS and send them some live samples, they check for most all diseases including Nosema if I remember correctly. how are you testing for mites?

 

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Ahhh yes, the highs and lows of beekeeping. Thanks for the brutally honest beekeeping story “soars”. Those who have been in it enough years have experienced plenty of both, though I think too often in these forums we tend to hear more of the success stories than we do from those who got frustrated and quit, or have gone out of business and never had it in them to post of their failures.
Without getting too wordy, allI I can suggest Is try to maintain a financial cushion from the good years, regroup, reload and carryon. Healthy bees require good forage and low mite numbers. The latter is somewhat beekeeper controllable, the former can be pretty fickle from one year to the next. In my experience, the financial allure of almond pollination can result in both the best and the worst of times.
 
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It was a really bad year for mites in California. The perfect weather during almond bloom gave varroa a real head start on the season.
You will likely find plenty of nosema spores in weak dwindling hives.
I think the consensus is that fumidil fed at the old rate for nosema apis is inadequate for n.c.
So look at a strong drench instead.
 

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Post 3 accurately describes the procedure for testing. They are hard to count and I dont think counting is all that accurate anyways. You will know a big load of spores when you see it! When you have dwindling hives right now, and nosema is showing up in lots of the samples, what do you have to lose by treating them? You will never know with certainty that nosema is the cause. But its sure a suspect.
Though I still believe Varroa is the root of all evil.
 

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Discussion Starter #12 (Edited)
Gentlemen, thank you for your replies! Ok, let me answer your questions...

We treat with oxilic acid using the ProVap 110 Sideliner/Commercial Oxalic Acid Vaporizer in late spring, then once again in late summer or fall. We treat every 5-7 days and give 4-5 treatments. I am convinced it cannot be a Varroa mite problem because 3-4 years ago, we allowed the mite load to go to high and began to see the deformed wing virus. We also saw massive mite numbers on the SBB's after vaporization [100-300 mites at a time]. I realize the alcohol and sugar roll is the golden standard to test for mite load. The majority of our colonies have sticky bottom boards and it is easy to monitor for mites with the these boards. I will be honest with you...after OA vapor treatment, for months we have a near zero mite load....we simply do not permit the mites loads to get any where even near a problem level during any time of the year. So we treat for mites twice a year, even if the mite levels are not very high.

Last winter we went from 200 colonies down to 4. Last spring, we jumped from 4 colonies back up to 126. We did not catch many swarms, but did lots of splits and queen grafting. I was hoping to be able to survive with at least 50-100 colonies and take them to the almond orchards. My goal is to at least regain my original investment.

After recently reading Randy Oliver's articles on Nosema, I purchased a microscope two weeks ago. Today, JRG is coming over to help me read the bees/spore counts under the microscope. So, no, I did not wait for the spore count to begin treatment. I began treatment two days ago by diluting a 25 gram bottle of Fumidil-B in 5 gallons of room temperature sugar syrup. I placed half gallon bottles on a number of colonies. I am presently waiting for the arrival of a 500 gram bottle of Fumidil-B that I purchased a week ago.

Randy indicated the greatest successes on bee nosema c survival was achieved by mixing the Fumidil-B in pollen sub! I plan on treating all colonies using the sugar syrup first, pollen sub next.

I am hoping to take 10-20 colonies to the almond orchards so I can at least recoup my costs from last year.

Please let me know if there are any other questions that might help discover the reason for our terrible losses.
 

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Correct, Logger Mike , Post #3 describes how to do the test, but the Million dollars was to "accurately quantify nosema spores under a microscope? ". No money offered for how to do the test.

That aside, I would suggest treating half the hives with Fumigilin. Back in "Aught Six", we saw sumptoms of dry first days larvae that cleaned up with treatment once. However, if the OP has shiny larvae, I would NOT expect to see a difference.

If you follow the time line, I would surmise that initially the OP had all new comb, and that the .problem arose ONLY after the comb had been used for a few seasons.

I would therefore suggest a four way test, Shake half the bees on new foundation, give half of those fumigilin. Of the remaining half on old comb, again give half Fumigilin.

Remember, the devils in the details of recording. Please report back what you do/find.

Crazy Roland
 

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To answer your original question to properly quantify the number of spores per bee is to perform a serial dilution. But as mentioned earlier, its prolly best to calculate a percentage of infection.
 

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Discussion Starter #15
We just finished the testing...

We tested in a variety of ways using various samples.

In the 50 honeybee crushed version we discovered approximately 15 spores in the FOV @ 400X. I think we are to multiply that by 50,000 and this gives us a 750,000 spore count.

What does this tell us?

Have you any other suggestions?

Thanks,

Soar
 

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Field of view 15 spores would approximate to 3 million spores. Not a lot but the next sample could be through the roof.
 

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Roland,

Thank you for your kindness in asking. This is a very painful story for me...

We started about 5 years ago after reading about bee colony collapse. I asked my wife if we should maybe start to keep honey bees [we have an orchard and live on a ranch]. We did a simple prayer: God, what are your thoughts on us getting honeybees? The very next day we came home and were shocked and amazed to find a monster swarm just a few feet from our front door. I began to do massive reading and asking questions. Within a year our numbers went vertical [we were catching 5-10 swarms a day during the peak of the swarm season]. For a two year period we were catching swarms faster than we could build the wood-ware. We also learned to make splits, graft queens, etc. So we began to experienced radical exponential growth. Two years ago I placed an ad on Craigslist to rent our honey bees to almond growers. I was thinking no one would call us. I was shocked to discover massive calls and people desperate for pollinators. So we began to take our honeybees 10 minutes away to the almond orchards...

Then the nightmare began. Last year, we went from 200 colonies all the way down to 4 colonies...so I think that is a 98% loss and I was crushed beyond words.

No one could give me a clear answer as to why. I have cared so much for the honeybees and most years we kept growing more and more...and it was one of the most wonderful endeavors we have ever done. It was very hard work [building boxes, lids, bottoms, bee stands, frames, etc.] and now I realize very expensive [I put my entire life savings into it].

Last winter, for the first time, we began to experience great losses. And I would be so happy to discover what it could be so I could rectify the problem. After losing 98% last winter I wanted to quit and get completely out of beekeeping because my dream was so utterly crushed I experienced a level of depression and emotional pain that I have never had. But after placing everything up for sale, it just did not feel right on the inside [I had no peace] so I only sold pallet fulls of unassembled boxes and frames and kept a few hundred boxes, lids and bottoms.

Here's some of the background:

We treat with oxilic acid using the ProVap 110 Sideliner/Commercial Oxalic Acid Vaporizer. To be honest with you, we only treat once or twice a year because for most of the year the Varroa mite load is nearly non existent. So I believe the problem cannot be the Varroa mite because we never permit the load levels to go high.

Two falls ago, for the first time ever, we began to see honey bees crawling all over on the ground within 100 feet of our bee yard. Last fall they did the same thing and they were dying.

The symptoms are the bee numbers begin to drop slowly in the late fall and continue to drop until the there is not one honeybee left in the box. The boxes are still filled with honey and pollen but not one darn honeybee inside! There is no outward sign of disease [example: no bad smell from EFB/AFB, no massive wax moth activity, no massive bee poop from Nosema apis, the honeybees simply begin to decline in numbers slowly but surely until not even one bee is left. We feed with sugar syrup and Mann Lake Ultra Bee Pollen.

This year, I stumbled upon Randy Oliver's description of Nosema ceranae. What stood out to me was that he said there were no outward signs of a problem...but there was a continual decline in bee population.

No one has been able to help me discover what is causing our honey bees to die in such massive numbers.

I thought I would buy a microscope and see if Nosema ceranae is the culprit.

I probably should not have done it, but I already began the treatment yesterday with the 12 strongest of our surviving colonies. This winter, we went from 126 colonies down to less than 50, so the losses are not as bad as last year, but there is still something terribly wrong because historically we have over wintered here with great success.

Ok, can someone please help us?

Thanks,

Soar
You are walking on the wrong trail ... I suffered the same losses many years ago ... I supposed to be nosemose cerenae as well ...but it was a wrong treatment of varroas... You supposed you treated well with oxilic acid ... but you were wrong ... try amitraz ! my poor advice ...
 
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