In vitro germination of Nosema apis spores under conditions favorable for the generation and maintenance of sporoplasms

Olsen, P. E.; Rice, W. A.; Liu, T.P.

doi:10.1016/0022-2011(86)90164-3

Cited by 26 publications

(17 citation statements)

References 9 publications

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“…Infection was initialized, i.e. germination of spores was triggered, by adding 100 µl cell suspension (∼2E+5 cells) and 400 µl 0.1 M sucrose in PBS‐buffer to the dried spores (see above) (Olsen et al ., 1986; Gisder et al ., 2010). The spore‐cell‐suspension was incubated at room temperature for 5 min and then transferred to 25 cm 2 tissue culture flasks (Greiner) with 4.5 ml TC‐100 medium supplemented with 11% FCS, 250 µg ml −1 penicillin/streptomycin (Roth) and 125 µl antimycotic/antibiotic solution (Sigma‐Aldrich).…”

Section: Methodsmentioning

confidence: 99%

A cell culture model for Nosema ceranae and Nosema apis allows new insights into the life cycle of these important honey bee‐pathogenic microsporidia

Gisder

Möckel

Linde

et al. 2010

Environmental Microbiology

106

100

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The population of managed honey bees has been dramatically declining in the recent past in many regions of the world. Consensus now seems to be that pathogens and parasites (e.g. the ectoparasitic mite Varroa destructor, the microsporidium Nosema ceranae and viruses) play a major role in this demise. However, little is known about host-pathogen interactions for bee pathogens and attempts to develop novel strategies to combat bee diseases have been hampered by this gap in our knowledge. One reason for this dire situation is the complete lack of cell cultures for the propagation and study of bee pathogens. Here we present a cell culture model for two honey bee-pathogenic microsporidian species, Nosema apis and N. ceranae. Our cell culture system is based on a lepidopteran cell line, which proved to be susceptible to infection by both N. ceranae and N. apis and enabled us to illustrate the entire life cycle of these microsporidia. We observed hitherto undescribed spindle-shaped meronts and confirmed our findings in infected bees. Our cell culture model provides a previously unavailable means to explore the nature of interactions between the honey bee and its pathogen complex at a mechanistic level and will allow the development of novel treatment strategies.

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Section: Methodsmentioning

confidence: 99%

A cell culture model for Nosema ceranae and Nosema apis allows new insights into the life cycle of these important honey bee‐pathogenic microsporidia

Gisder

Möckel

Linde

et al. 2010

Environmental Microbiology

106

100

View full text Add to dashboard Cite

show abstract

“…Possibly, older bees are less vulnerable to infection than younger bees. Also, we suspect that the method of feeding spores in sucrose solution inhibits spore germination in the ventriculus, because the osmotic shock required for germination (Olsen, 1986;De Graaf et al, 1993) cannot happen reliably. A dry preparation of spores would probably be a more effective inoculum because liquid food already in the ventriculus would then provide the osmotic shock.…”

Section: Discussionmentioning

confidence: 99%

Nosema apis infection in worker and queen Apis mellifera

et al. 2004

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-Worker and queen honey bees were fed individually with Nosema apis spores in sucrose solution and then returned to cages containing several hundred of their worker bee nestmates. After 3 to 7 days, the workers and queens that had been fed spores were sacrificed. Worker and queen ventriculi were removed and examined for spores by light microscopy, and DNA was extracted. The DNA was subjected to amplification with polymerase chain reaction, using primer sequences specific to N. apis DNA. The PCR analysis was more sensitive than examination for spores by light microscopy, in detecting N. apis infection. Worker bees and queen bees were infected at similar rates by the inoculation procedure.Apis mellifera / Nosema apis / PCR / queen / worker

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“…Several microsporidia have less dependence on the pH of the environment and have demonstrated spore discharge at both acidic and alkaline conditions (Hashimoto et al 1976;Undeen 1983;Undeen & Avery 1988b). In some species, dehydration by drying or hyperosmotic solutions followed by rehydration has been effective in promoting spore discharge (Gibbs 1953;Kramer 1960;Olsen et al 1986). In other microsporidia, dehydration followed by rehydration at an alkaline pH was effective (Undeen 1978;Undeen & Epsky 1990;Whitlock & Johnson 1990).…”

Section: Glugea Hertwigimentioning

confidence: 99%

“…10.10). It has been reported that the polar tube diameter can increase from 0.1 to 6 μm during sporoplasm passage (Lom & Vavra 1963;Ohshima 1966;Ishihara 1968;Weidner 1972Weidner , 1976Olsen et al 1986). It has been reported that the polar tube diameter can increase from 0.1 to 6 μm during sporoplasm passage (Lom & Vavra 1963;Ohshima 1966;Ishihara 1968;Weidner 1972Weidner , 1976Olsen et al 1986).…”

Section: Spore Germination: Polar Tube Dischargementioning

confidence: 99%

“…However, the mechanism(s) of activation and tube formation during discharge is still not completely resolved and both molecular, cellular biology and ultrastructural questions remain about the functioning, assembly, composition, and three-dimensional (3D) structure of the polar tube and the invasion process. 1963;Ishihara 1968;Weidner 1972Weidner , 1976Olsen et al 1986), and its length shortens by 5-10% after sporoplasm discharge (Frixione et al 1992). Diagram of a microsporidian spore.…”

Section: Introductionmentioning

confidence: 99%

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