Identification of Candidate Agents Active against N. ceranae Infection in Honey Bees:
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  1. #1
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    Default Identification of Candidate Agents Active against N. ceranae Infection in Honey Bees:

    Identification of Candidate Agents Active against N. ceranae Infection in Honey Bees: Establishment of a Medium Throughput Screening Assay Based on N. ceranae Infected Cultured Cells
    Many flowering plants in both natural ecosytems and agriculture are dependent on insect pollination for fruit set and seed production. Managed honey bees (Apis mellifera) and wild bees are key pollinators providing this indispensable eco- and agrosystem service. Like all other organisms, bees are attacked by numerous pathogens and parasites. Nosema apis is a honey bee pathogenic microsporidium which is widely distributed in honey bee populations without causing much harm. Its congener Nosema ceranae was originally described as pathogen of the Eastern honey bee (Apis cerana) but jumped host from A. cerana to A. mellifera about 20 years ago and spilled over from A. mellifera to Bombus spp. quite recently. N. ceranae is now considered a deadly emerging parasite of both Western honey bees and bumblebees. Hence, novel and sustainable treatment strategies against N. ceranae are urgently needed to protect honey and wild bees. We here present the development of an in vitro medium throughput screening assay for the identification of candidate agents active against N. ceranae infections. This novel assay is based on our recently developed cell culture model for N. ceranae and coupled with an RT-PCR-ELISA protocol for quantification of N. ceranae in infected cells. The assay has been adapted to the 96-well microplate format to allow automated analysis. Several substances with known (fumagillin) or presumed (surfactin) or no (paromomycin) activity against N. ceranae were tested as well as substances for which no data concerning N. ceranae inhibition existed. While fumagillin and two nitroimidazoles (metronidazole, tinidazole) totally inhibited N. ceranae proliferation, all other test substances were inactive. In summary, the assay proved suitable for substance screening and demonstrated the activity of two synthetic antibiotics against N. ceranae.
    http://journals.plos.org/plosone/art...l.pone.0117200
    americasbeekeeper.com
    [email protected]

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  3. #2
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    Default Re: Identification of Candidate Agents Active against N. ceranae Infection in Honey B

    How were the treatments administered? ( my phone does not pull up that file )

  4. #3
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    Default Re: Identification of Candidate Agents Active against N. ceranae Infection in Honey B

    The assay is based on our recently developed cell culture model for Nosema spp., the lepidopteran cell line IPL-LD 65Y. This cell line originating from Lymantria dispar has been shown to be susceptible to N. ceranae infection and to support the entire life cycle of N. ceranae [20]. We combined the cell culture assay with quantitative detection of N. ceranae in infected cells via an RT-PCR-ELISA (reverse transcriptase-polymerase chain reaction-enzyme linked immunosorbent assay) protocol and adopted the assay for the 96-well microplate format to enable automated analysis. We first verified our test system using fumagillin as positive control, paromomycin as negative control, and clioquinol as a highly cytotoxic agent, to cover the possible range of effects on both cells and microsporidia. We then tested several commercially available agents for cytotoxicity and inhibition of N. ceranae intracellular development and demonstrate the efficacy of the synthetic antibiotics metronidazole and tinidazole against N. ceranae.
    americasbeekeeper.com
    [email protected]

  5. #4
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    Default Re: Identification of Candidate Agents Active against N. ceranae Infection in Honey B

    So, the summary, for those without PhDs, is that Fumagilin-B, metronidazole, and tinidazole are effective treatments against Nosema?
    If you want to be successful, study successful people and do what they do.
    Zone 4a/b

  6. #5
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    Default Re: Identification of Candidate Agents Active against N. ceranae Infection in Honey B

    does it say how the treatments are administered? drench, through syrup feeding, dusting?
    Ian Steppler >> Canadian Beekeeper's Blog
    www.stepplerfarms.com

  7. #6
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    Default Re: Identification of Candidate Agents Active against N. ceranae Infection in Honey B

    Based on my [limited] understanding of Nosema, I would think that it needs to be consumed, like mixing the Fumagilin in syrup as I currently do. I don't know about the 'zoles? Could they be dusted on the frames like Teramycin?
    If you want to be successful, study successful people and do what they do.
    Zone 4a/b

  8. #7
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    Default Re: Identification of Candidate Agents Active against N. ceranae Infection in Honey B

    what has hooked my interest lately is the method of application. Does feeding through the syrup actually target the nosema adequately? Drench method, done at what doses at how many intervals? feeding though patties, dusting over frames with icing sugar... and so on...

    AmericasBeekeeper's link provides some technical talk which I think is suggesting fumagillin's efficacy . How are they applying it to get that efficacy?
    Ian Steppler >> Canadian Beekeeper's Blog
    www.stepplerfarms.com

  9. #8
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    Default Re: Identification of Candidate Agents Active against N. ceranae Infection in Honey B

    Quote Originally Posted by Ian View Post
    what has hooked my interest lately is the method of application. Does feeding through the syrup actually target the nosema adequately? Drench method, done at what doses at how many intervals? feeding though patties, dusting over frames with icing sugar... and so on...

    AmericasBeekeeper's link provides some technical talk which I think is suggesting fumagillin's efficacy . How are they applying it to get that efficacy?
    The instructions I have in front of me for the Fumagilin is 5g in a gallon of syrup for a 2 chamber colony, done twice. I use a reduced quantity of syrup- 5g in a quart, which I put in a zip-lock baggie on top of the frames in the top box. Once the baggie is on top of the frames, I carefully slit the top side of it with a sharp knife. The bees usually consume it fairly rapidly. When they are finished with the first dose, they get the second dose in the same manner.

    I had at least one hive that did not get the full treatment this fall, and there was evidence of Nosema. The properly treated hives seemed OK. That is by no means a proper experiment, but...in general, my results with the Fumagilin over the last several years (and the dramatic effect of the untreated hive) lead me to believe that this is adequately effective.
    If you want to be successful, study successful people and do what they do.
    Zone 4a/b

  10. #9
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    Default Re: Identification of Candidate Agents Active against N. ceranae Infection in Honey B

    Quote Originally Posted by Ian View Post
    does it say how the treatments are administered? drench, through syrup feeding, dusting?
    No Ian. This study was in vitro, not in vivo. I will summarize what I had understood trought this study and other two papers:
    There are three distinct phases in the development of microsporidia: the infective (environmental) phase represented by mature spores, the proliferative phase (merogony) responsible for the massive increase in number of the parasite, and the sporogonic phase (sporogony), in which sporonts produce sporoblasts that mature into spores.

    Previous studies had shown that 72 hours after infection merogony is nearly completed and sporogony, i.e. the development of new spores marking the final phase of the vegetative cycle, is initiated [20].

    The substances to be tested were selected based on their known inhibitory effect on other microsporidia, fungi or intracellular parasites. We tested the synthetic antibiotics
    1) albendazole,
    2) ornidazole,
    3 and 4) tinidazole, and metronidazole
    because both in vitro and in vivo studies have demonstrated that benzimidazoles and nitroimidazoles and its derivatives have activity against microsporidia in vertebrates as well as in invertebrates [67–71]


    Conclusions:
    1)Therefore, it was not possible to cure infected cells with non-cytotoxic concentrations of albendazole
    2) Therefore, ornidazole like albendazole has no significant in vitro activity against N. ceranae.
    3 and 4) These results indicated that at a concentration of 2 mg/ml both tinidazole and metronidazole can completely inhibit N. ceranae infection of IPL-LD 65Y cells and, therefore, were as effective as fumagillin in vitro. However, both substances will hardly have a future as anti-nosemosis drugs in honey bees. Nitroimidazoles are antibiotics and the use of antibiotics in honey bee colonies and in particular the use of nitroimidazoles in food animals and animal food is prohibited in many countries [73] including the European Union.

    I hope I have not distorted nothing that can not be replaced with a more accurate reading than I could accomplish.

  11. #10
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    Default Re: Identification of Candidate Agents Active against N. ceranae Infection in Honey B

    I read the study quickly and with no PHD it was a tough read, but I didn't think being effective killing nosema was the issue. Isn't it the bounce back that occurs the issue? It's like us taking half the recommended dose of antibiotics and then having the bug come back roaring? This is why I stopped using Fumagilin. It's expensive, so it needs to have a long term benefit for me to use it. I'm sure I'm oversimplifying this, but barring the obvious Apis signs I won't be using it unless I hear it is of real benefit on my small scale.
    Dan Boylan, When in doubt "It's mites".

  12. #11
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    Default Re: Identification of Candidate Agents Active against N. ceranae Infection in Honey B

    My questions are in regard to using Fumagilin as a drench. I know a couple of the commercial beekeepers in the southern region are doing go and they credit its use for an increase in bee health. I would be interested in hearing from anyone who is "spraying" the bees.
    Charlotte Anderson - SC Master Beekeeper
    Carolina Honeybees

  13. #12
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    Default Re: Identification of Candidate Agents Active against N. ceranae Infection in Honey B

    David
    beebotanical.com 40 years-4000 colonies-treatment eo's

  14. #13
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    Default Re: Identification of Candidate Agents Active against N. ceranae Infection in Honey B

    Quote Originally Posted by DPBsbees View Post
    I stopped using Fumagilin. It's expensive, so it needs to have a long term benefit for me to use it. I'm sure I'm oversimplifying this, but barring the obvious Apis signs I won't be using it unless I hear it is of real benefit on my small scale.
    My unplanned experiment (bad beekeeping) has convinced me that prophylactic treatment with Fumagilin is worth the expense. As of yesterday, treated hives doing well, untreated showing signs of Nosema, at least two dead and one a nasty mess. I'll spend the money, dead hives don't make honey.
    If you want to be successful, study successful people and do what they do.
    Zone 4a/b

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