Long-term storage of Ascosphaera aggregata and Ascosphaera apis, pathogens of the leafcutting bee (Megachile rotundata) and the honey bee (Apis mellifera)

Jensen, Annette Bruun; James, Rosalind R.; Eilenberg, Jørgen

doi:10.1016/j.jip.2009.03.004

Cited by 10 publications

(7 citation statements)

References 16 publications

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“…Once new mycelia were observed, mating tests with the reference strains ARSEF 7405 and 7406 were performed. All the isolates were stored in 20% glycerol at −80 °C (as described in Jensen et al ., ).…”

Section: Methodsmentioning

confidence: 97%

Polymorphic DNA sequences of the fungal honey bee pathogen Ascosphaera apis

Jensen

Welker

Kryger³

et al. 2012

FEMS Microbiol Lett

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The pathogenic fungus Ascosphaera apis is ubiquitous in honey bee populations. We used the draft genome assembly of this pathogen to search for polymorphic intergenic loci that could be used to differentiate haplotypes. Primers were developed for five such loci, and the species specificities were verified using DNA from nine closely related species. The sequence variation was compared among 12 A. apis isolates at each of these loci, and two additional loci, the internal transcribed spacer of the ribosomal RNA (ITS) and a variable part of the elongation factor 1α (Ef1α). The degree of variation was then compared among the different loci, and three were found to have the greatest detection power for identifying A. apis haplotypes. The described loci can help to resolve strain differences and population genetic structures, to elucidate host-pathogen interaction and to test evolutionary hypotheses for the world's most important pollinator: the honey bee and one of its most common pathogens.

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Section: Methodsmentioning

confidence: 97%

Polymorphic DNA sequences of the fungal honey bee pathogen Ascosphaera apis

Jensen

Welker

Kryger³

et al. 2012

FEMS Microbiol Lett

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“…In addition, A. apis isolates, as seen for many other microbes, can lose virulence when kept in culture with repeated transfers, thus it is important to utilize a long term storage method soon after an isolate is obtained. Two different culture preservation methods have been successfully used: cryopreservation (Jensen et al ., 2009a) and propagation on Integral Rice Kernels (IRK) (Palacio et al ., 2007). Contamination during storage seems to be less risky during cryopreservation at −80°C, however for larger quantities and for laboratories not equipped with cryopreservation capacity, the use of rice kernels is a great alterative.…”

Section: Chalkbroodmentioning

confidence: 99%

Standard methods for fungal brood disease research

Jensen

Aronstein²,

Flores

et al. 2013

Journal of Apicultural Research

146

130

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Summary Chalkbrood and stonebrood are two fungal diseases associated with honey bee brood. Chalkbrood, caused by Ascosphaera apis, is a common and widespread disease that can result in severe reduction of emerging worker bees and thus overall colony productivity. Stonebrood is caused by Aspergillus spp. that are rarely observed, so the impact on colony health is not very well understood. A major concern with the presence of Aspergillus in honey bees is the production of airborne conidia, which can lead to allergic bronchopulmonary aspergillosis, pulmonary aspergilloma, or even invasive aspergillosis in lung tissues upon inhalation by humans. In the current chapter we describe the honey bee disease symptoms of these fungal pathogens. In addition, we provide research methodologies and protocols for isolating and culturing, in vivo and in vitro assays that are commonly used to study these host pathogen interactions. We give guidelines on the preferred methods used in current research and the application of molecular techniques. We have added photographs, drawings and illustrations to assist bee-extension personnel and bee scientists in the control of these two diseases.

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“…Storage conditions were tested using A. apis spores and hyphae (Jensen et al, 2009). Both freeze-dried and cryogenically stored A. apis spores preserved well.…”

Section: Fungal Culture and Storage Conditionsmentioning

confidence: 99%