Evaluation of <i>Nosema ceranae</i> spore-specific polyclonal antibodies

Aronstein, Katherine A.; Saldivar, Eduardo; Webster, Thomas C.

doi:10.3896/ibra.1.50.2.06

Cited by 7 publications

(6 citation statements)

References 17 publications

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“…Diagnostic assays that establish the presence/absence of disease may provide limited information about the overall health of a colony [53], [54]. Currently, the standard method to diagnosis Nosema spp.…”

Section: Discussionmentioning

confidence: 99%

Physiological and Behavioral Changes in Honey Bees (Apis mellifera) Induced by Nosema ceranae Infection

2013

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Persistent exposure to mite pests, poor nutrition, pesticides, and pathogens threaten honey bee survival. In healthy colonies, the interaction of the yolk precursor protein, vitellogenin (Vg), and endocrine factor, juvenile hormone (JH), functions as a pacemaker driving the sequence of behaviors that workers perform throughout their lives. Young bees perform nursing duties within the hive and have high Vg and low JH; as older bees transition to foraging, this trend reverses. Pathogens and parasites can alter this regulatory network. For example, infection with the microsporidian, Nosema apis, has been shown to advance behavioral maturation in workers. We investigated the effects of infection with a recent honey bee pathogen on physiological factors underlying the division of labor in workers. Bees infected with N. ceranae were nearly twice as likely to engage in precocious foraging and lived 9 days less, on average, compared to controls. We also show that Vg transcript was low, while JH titer spiked, in infected nurse-aged bees in cages. This pattern of expression is atypical and the reverse of what would be expected for healthy, non-infected bees. Disruption of the basic underpinnings of temporal polyethism due to infection may be a contributing factor to recent high colony mortality, as workers may lose flexibility in their response to colony demands.

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Section: Discussionmentioning

confidence: 99%

Physiological and Behavioral Changes in Honey Bees (Apis mellifera) Induced by Nosema ceranae Infection

2013

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“…1B, 6a). It is also important to point out that this new ELISA detects only environmental spores of N. ceranae (Aronstein et al 2011). Because various developmental stages of Nosema are simultaneously found in the host, we expected that qRT-PCR would produce higher counts than those measured by ELISA (Fig.…”

Section: Discussionmentioning

confidence: 99%

“…DQ486027) as described in the study by Chen et al (2009). The second primer set was designed to amplify a 737-bp fragment of N. ceranae spore wall protein SWP-32 gene that amplifies the same region of DNA used for raising GAT antibody (Aronstein et al 2011). PCR consisted of 1Á25 U of GoTaq ® Flexi DNA polymerase (Promega Co., Madison, WI, USA) with the colourless 59 GoTaq ® Flexi buffer, 0Á20 mmol l À1 dNTP mix, 2Á5 mmol l À1 MgCl 2 , 0Á125 lmol l À1 of each primer, 0Á75 ll of 10009 dilution of SYBR Green I and 1 ll of DNA.…”

Section: Quantitative Real-time Pcr (Qrt-pcr)mentioning

confidence: 99%

“…The lowest volume spore preparations tested in this study was 1Á66 ll per replicate or 5 ll per sample. Samples were diluted to 50 ll with double distilled autoclaved water and prepared as described previously (Aronstein et al 2011). The Express ELISA Kit for Rabbit Primary Antibodies (GenScript, Piscataway, NJ, USA) was used according to the manufacture's protocol to test the spore suspensions.…”

Section: Enzyme-linked Immunosorbent Assaymentioning

confidence: 99%

“…In this study, we describe a serological approach to N. ceranae detection using a highly specific genomic antibody developed against N. ceranae SWP-32 spore antigen by Aronstein et al (2011). The advantage of a serological method is that it can be easily adopted for both laboratory and field applications.…”

Section: Introductionmentioning

confidence: 99%

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A serological method for detection of Nosema ceranae

2012

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Aims: We developed a new method for detection of the intracellular parasite, Nosema ceranae, one of the most economically devastating pathogens of the honeybee. Methods and Results: The SWP-32 antibody was used for the development of an enzyme-linked immunosorbent assay (ELISA). We also compared the efficiency of this ELISA to microscopy and quantitative real-time (qRT) PCR, the methods currently in use. Conclusions: ELISA is comparable in sensitivity with the qRT-PCR, less expensive and faster. When this method is commercialized and made available to bee-keepers, it will allow them to make informed decisions for the application of in-hive chemicals. Hence, bee-keepers may be able to determine when treatments for control of N. ceranae are unnecessary and reduce the cost, time and possible side effects of these treatments. Significance and Impact of the Study: This assay provides the first serological method for detection of N. ceranae in bee colonies, which is as sensitive as DNA amplification. It can be easily adopted for both laboratory and field applications.

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