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Author Topic: AFB spores  (Read 5416 times)
Mici
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« on: November 22, 2007, 08:18:48 AM »

HI!

ummm, i really can't find a picture of AFB spores under a microscope, now...if anyone know of one, i'd be happy if you would post a link, or send me that picture, also, information about magnification of that picture is important.

thank you for your help
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Jerrymac
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« Reply #1 on: November 22, 2007, 09:22:21 AM »

http://www.teagasc.ie/oakpark/bru/bru-foulbrood.htm
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Mici
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« Reply #2 on: November 22, 2007, 12:37:34 PM »

thank you!
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Finsky
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« Reply #3 on: November 22, 2007, 02:58:32 PM »

HI!

ummm,

Hmm, where you need hose spores?

When spores are calculated they boil first example. Spores stay alive and then stuff will be  cultivated on certain substrate.


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Mici
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« Reply #4 on: November 22, 2007, 03:30:31 PM »

oh, i don't need them, he-he
someone asked a few questions about micro scoping and that he'd like to try it, and maybe test for AFB, so....I'm trying to help, to find a good picture of AFB spores through microscope.
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Finsky
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« Reply #5 on: November 23, 2007, 01:02:09 AM »

oh, i don't need them, he-he
someone asked a few questions about micro scoping and that he'd like to try it, and maybe test for AFB, so....I'm trying to help, to find a good picture of AFB spores through microscope.

AFB cannot be found by microscope. It is found wih laboratorium cultivation and by that, it stands 130C temperature.
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Mici
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« Reply #6 on: November 23, 2007, 08:59:39 AM »

i'm not sure if we understand each other.
around here, anyone willing and in doubt, can get his honey tested for AFB presency.
so...many say AFB is always present so there are no negative results (although, if number of spores per some ammount does not exceed certain value, they declare that sample as negative)

so...what do they do with that honey? i mean...i can't imagine any other way than to look at the honey under a microscope.

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taipantoo
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« Reply #7 on: November 23, 2007, 09:07:42 AM »

oh, i don't need them, he-he
someone asked a few questions about micro scoping and that he'd like to try it, and maybe test for AFB, so....I'm trying to help, to find a good picture of AFB spores through microscope.

AFB cannot be found by microscope. It is found wih laboratorium cultivation and by that, it stands 130C temperature.

I don't understand your statement.
Why can't you see AFB spores with a microscope?

When collecting mushrooms, the only way to definitely know a safe mushroom from a poison/toxic look-a-like is to look at the spores.
My books say that 400 power is sufficient.

Am I missing something here?
Tai
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Finsky
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« Reply #8 on: November 23, 2007, 11:13:14 AM »

Spores are cultivated in laboratory and spores are calculated via bacterium clusters -like other bacteria cultivations
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Cindi
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« Reply #9 on: November 24, 2007, 09:31:51 AM »

Finksy, yes the spores are cultivated, but one has to use a microscope to see them, they are not visible by the naked eye, or am I missing something too.  Have a wonderful and beautiful day.  Cindi
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« Reply #10 on: November 25, 2007, 04:05:28 PM »

Finksy, yes the spores are cultivated, but one has to use a microscope to see them, they are not visible by the naked eye, or am I missing something too.  Have a wonderful and beautiful day.  Cindi


Do you mean nosema?

AFB goes this way and are visible in petri tray

http://www.apimondia.org/apiacta/articles/2003/ritter_1.pdf

I cannot notice microscope in method.

*******************
APIACTA 41 (2006) PAGE 99-109 99
Monitoring for American Foulbrood Spores from Honey and Bee Samples
in Canada

http://www.apimondia.org/apiacta/articles/2006/pernal_1.pdf

.
*****************

Sample is heated in 90C temperature that other bacterias will die

Samlpe is cultivated petri tray on nutrition where bacterias make colonies.
Sample must be diluted so that each spore makes one indivudual colony on nutrition substrate.


.
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rdy-b
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« Reply #11 on: November 25, 2007, 06:15:58 PM »

Finksy, yes the spores are cultivated, but one has to use a microscope to see them, they are not visible by the naked eye, or am I missing something too.  Have a wonderful and beautiful day.  Cindi


Do you mean nosema?

AFB goes this way and are visible in petri tray

http://www.apimondia.org/apiacta/articles/2003/ritter_1.pdf

I cannot notice microscope in method.

*******************
APIACTA 41 (2006) PAGE 99-109 99
Monitoring for American Foulbrood Spores from Honey and Bee Samples
in Canada

http://www.apimondia.org/apiacta/articles/2006/pernal_1.pdf

.
*****************

Sample is heated in 90C temperature that other bacterias will die

Samlpe is cultivated petri tray on nutrition where bacterias make colonies.
Sample must be diluted so that each spore makes one indivudual colony on nutrition substrate.


.


wonder what they mean on page 126 when they say-(determined by a culture counting device)
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Cindi
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« Reply #12 on: November 25, 2007, 10:28:52 PM »

Finsky, I am standing corrected.  The paper indicates that the paeninbacillus larvae larvae colonies are visible in the petri dish as white and have a concave form and rough surface.  I think I misunderstood something.

So AFB colonies are visible, but is a microscope required to see the spores?  I didn't read in that paper anything about that part. 

There is another test called the Holtz milk test that can be performed as well to test for AFB.  Can't recall an awful lot about this specific test, but it is a viable one for a quick ident on AFB.  Beautiful day, great life.  Cindi
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There are strange things done in the midnight sun by the men who moil for gold.  The Arctic trails have their secret tales that would make your blood run cold.  The Northern Lights have seen queer sights, but the queerest they ever did see, what the night on the marge of Lake Lebarge, I cremated Sam McGee.  Robert Service
rdy-b
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« Reply #13 on: November 25, 2007, 11:17:11 PM »

 cool Smiley http://www.agf.gov.bc.ca/apiculture/factsheets/205_disdetect.htm  I think most pepole use a microscope   to aid in  Diagnosis

AFB is caused by Paenibacillus larvae , a spore-forming bacterium.
A microscope slide can be prepared by dissolving a small part of an AFB scale. Stir the scale with a toothpick in a droplet of water placed on a slide and apply a cover slip.
Under 400X magnification, the AFB spores are readily visible. AFB spores are characterized by being very slightly oblong, uniform in size and shape. The spores “jiggle” in a characteristic Brownian movement.
P. larvae is competitive and does not tolerate growth of other bacteria in the parasitized bee larva. As a result, most microscopic slides will show a predominance of P. larvae spores. This is not always the case with poor samples or those left in the collection bag for too long. In such case, secondary invaders such as moulds, will appear.
Control and  Wink RDY-B
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Finsky
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« Reply #14 on: November 25, 2007, 11:44:20 PM »

cool Smiley http://www.agf.gov.bc.ca/apiculture/factsheets/205_disdetect.htm  I think most people use a microscope   to aid in  Diagnosis



Very strange idea in that "fact sheet".

* you identify first AFB with naked eye from scales, etc.
* then you make a spore sample from scale and indentify it? Makes no sense
* better forget that identification.

If you really want to see it with microscope, the good tool is really expensive.

.
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rdy-b
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« Reply #15 on: November 25, 2007, 11:58:35 PM »

cheap insurance with all the scare of nosemea crena  cheesy                                                                        http://cgi.ebay.com/ws/eBayISAPI.dll?ViewItem&item=180181057884&ssPageName=MERC     cheesy RDY-B
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taipantoo
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« Reply #16 on: November 26, 2007, 10:49:47 AM »

Finksy, yes the spores are cultivated, but one has to use a microscope to see them, they are not visible by the naked eye, or am I missing something too.  Have a wonderful and beautiful day.  Cindi


Do you mean nosema?

AFB goes this way and are visible in petri tray

http://www.apimondia.org/apiacta/articles/2003/ritter_1.pdf

This cultivation in a petri dish is the cultivation of pathogens that are found on the spores, not the cultivation of the spores.
We are talking about two entirely different things here.


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rdy-b
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« Reply #17 on: November 26, 2007, 08:57:15 PM »

taipantoo- AFB goes this way grin                                                                       http://www.agf.gov.bc.ca/apiculture/factsheets/205_disdetect.htm                                                                                                                        being able to check for nosemia is a big plus and goes this way                                          http://www.wildwoodlabs.com/viewer.php?article_id=84    cool  RDY-B
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Jerrymac
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« Reply #18 on: November 27, 2007, 10:20:39 AM »

He wants to look at it under a microscope. What is wrong with that?
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taipantoo
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« Reply #19 on: November 27, 2007, 01:07:50 PM »

It seemed to me that the discussion was leaning towards the cultivation of the spores.
Spores do not make new spores.
The cultivation of bacterium on the spores can lead to identification of spores because you can see a bacteria colony with the naked eye when you can't see the spores, hence, no microscope needed.

I was not aware that the bacteria was a spore forming bacteria, so, technically speaking, I can understand the term 'cultivation of spores' even though it is the 'cultivation of bacteria' that are producing the spores.

I apologize if I have caused any confusion and thanks for clearing this up for me.
I should have done more reading before posting.
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Finsky
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« Reply #20 on: November 27, 2007, 01:27:28 PM »



I apologize if I have caused any confusion

Never mind.  I have had bees 45 years but 5 years ago forum friends told me that AFB spores stand boiling temperature. I was sure that they die as other bacterias in over 70C temperature.


.
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tig
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« Reply #21 on: December 17, 2007, 05:55:45 AM »

when i have my colonies tested, i cut a section of the comb that contains sealed brood. ideally the brood is still in the soft larval stage not almost fully formed into pupa.  then i capture about 30 live bees and send them along to be tested.

the contents of the adult bee gut are smeared and examined under a microscope.  since it's the university that does the tests, i was fortunate to be able to examine the slide since they used a teaching microscope with a pointer.  my slides didn't have any signs of AFB spores, but i was shown a slide that did have the spores.  the spores look rod-like and slightly elongated.  the microbiologist explained to me that there is a type of spore which looks very similar and they call it the lactic type.  thats the good kind of bacteria.

it's not only the presence of the spores they look at, but also the number and movement. once they see suspicious spores, they start making a culture test and that takes some time because they have to grow it in a medium.  the culture test is the most definite and unarguable and thats when you start praying while waiting for the results LOL.
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Mici
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« Reply #22 on: December 17, 2007, 01:14:38 PM »

when i have my colonies tested, i cut a section of the comb that contains sealed brood. ideally the brood is still in the soft larval stage not almost fully formed into pupa.  then i capture about 30 live bees and send them along to be tested.

the contents of the adult bee gut are smeared and examined under a microscope.  since it's the university that does the tests, i was fortunate to be able to examine the slide since they used a teaching microscope with a pointer.  my slides didn't have any signs of AFB spores, but i was shown a slide that did have the spores.  the spores look rod-like and slightly elongated.  the microbiologist explained to me that there is a type of spore which looks very similar and they call it the lactic type.  thats the good kind of bacteria.

it's not only the presence of the spores they look at, but also the number and movement. once they see suspicious spores, they start making a culture test and that takes some time because they have to grow it in a medium.  the culture test is the most definite and unarguable and thats when you start praying while waiting for the results LOL.

best info so far, thank you very much tig!
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tig
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« Reply #23 on: December 18, 2007, 03:58:52 AM »

you're welcome mici.  unfortunately i didn't ask the magnification of the teaching microscope.  i have my brood samples tested every 6 months for my peace of mind.  if your friend has access to a laboratory, maybe in a university, he can ask if they can test his sample and request that he be present.  remember, the microscope only shows what maybe "suspicious" looking spores. if the colony doesn't show any field signs of AFB,  you should proceed to have a culture done to remove any doubt.

last may when i had my colonies tested, they found a lot of "suspicious" and possibly AFB spores.  what made it bad was there was a great number present and very actively moving in each sample. to be on the safe side, i quarentined all the bees in that one apiary which showed the spores and waited for the culture test results. thankfully it turned out the be the lactic type which was good!  but i had a lot of sleepless nights and stressed filled days until the culture results came out LOL.
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tig
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« Reply #24 on: December 18, 2007, 05:26:22 AM »

i forgot to mention the rationale why they do the microscopic test on the adults first.  the premise is that if the colony does have AFB and the adult bees have a lot of the spores, then the brood will be overflowing with the spores since AFB is primarily a brood disease.  the culture test will be done on the brood once suspicious spores are found.
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Mici
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« Reply #25 on: December 18, 2007, 06:14:40 PM »

the college that was asking about it, was asking whether it's reasonable to buy a microscope for that matter.
however i do have access to my university microscopes, i've already arranged to go check my honey, just to see, what there's actually to see, just have to get some time...darn.

if it's a school microscope i'm sure it's no more than 10x40, i hope we get some nice days so i can get a fresh bee or two, hehe.
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tig
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« Reply #26 on: December 21, 2007, 08:18:49 PM »

hi mici,

    i checked with the university about the microscope needed.  you will be needing one with an oil emersion lens.  thats usually with a 1000 magnification.
gl with your test!
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