The use of a microinjection procedure for large-scale production of the microsporidian Nosema eurytremae in Pieris brassicae

Pilley, Barbara M.; Canning, Elizabeth U.; Hammond, J. C.

doi:10.1016/0022-2011(78)90200-8

Cited by 14 publications

(7 citation statements)

References 9 publications

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“…Spores of N. eurytremae were harvested from Pieris brassicae pupae after a micro-injection procedure described by Pilley, Canning & Hammond (1978). Spores of N. algerae were also derived from passage through P. brassicae (Lai & Canning, 1982).…”

Section: Parasitesmentioning

confidence: 99%

Culture of microsporidia from invertebrates in vertebrate cells

1982

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Nosema algerae and N. eurytremae were successfully cultured in embryonic rat brain, Xenopus XTC-6 and Chang liver cells, and in embryonic rat brain and XTC-6 cells respectively. No parasites grew in cultures incubated at 38 °C but development took place in cells incubated at 34 °C and 27 °C. Increased levels of infections of the cultured cells were achieved by centrifugation of the spores on to the cells. The level of infection was also related to the type of medium used for hatching the spores, modifies NMRI giving better results than Leibowitz L15, and to the interval between adding the spores to the cell cultures and centrifugation.

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Section: Parasitesmentioning

confidence: 99%

Culture of microsporidia from invertebrates in vertebrate cells

1982

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“…The high doses required to secure cercarial depression by N. eurytremae pose a problem in spore production for possible field trials. A start has been made towards Pathogenicity of a microsporidium in liver flukes 391 mass production by the development of a tissue culture system for this microsporidium (Higby et al 1979) and an efficient method for growth in laboratorybred Lepidoptera (Higby et al 1979;Pilley et al 1979). These methods require improvement, especially in the in vitro system, but demonstrate clearly that spore production is not dependent on growth of the parasite in its natural host.…”

Section: Discussionmentioning

confidence: 99%

“…The rediae were disrupted by grinding in the homogenizer and the spores decanted for immediate use or storage at 4 °C. Spores were also mass-produced by intrahaemocoelic inoculation into Pieris brassicae larvae and harvesting from pupae (Pilley, et al 1979).…”

Section: Preparation Of Spores Of N Eurytremaementioning

confidence: 99%

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An experimental study of the effects of Nosema eurytremae (Microsporida: Nosematidae) on the liver fluke Fasciola hepatica

1979

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SUMMARYA microsporidian parasite, Nosema eurytremae, was fed to Lymnaea trunculata infected with Fasciola hepatica. Microsporidian infection of the snail was always light, though spores were present in all tissues, but the rediae became heavily infected in the parenchyma. The proportion of infected rediae increased with time and was especially high in ageing infections. Although rediae were only lightly infected when examined before cercarial release began, the infections became progressively heavier, so that towards the end of the life of the snails a high proportion of rediae were totally destroyed and contained no cercariae. Cercarial output was significantly depressed over a long period of active shedding by snails which had been hyperinfected with 1 × 10 6 spores and some of the liberated cercariae were themselves infected.

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“…Hyperinfection of trematode larvae was achieved by allowing L. truncatula to browse on spore-contaminated algal plates (Canning et al 1979) and by placing the aquatic snails individually in the wells of tissue-culture trays to feed on spore-contaminated alginate discs (Lai & Canning, 1980). Spores purified from the naturally hyperinfected L. peregra or from the experimentally infected L. truncatula or P. planorbis were used in attempts to infect non-trematode hosts, (a) Third and 4th instar Pieris brassicae larvae were inoculated with spores into the haemocoele (Pilley, Canning & Hammond, 1978) or were fed with spores by pushing the head sharply down on to a small drop of spore suspension, so that the drop was automatically sucked back into the mouth. (b) First to 3rd instars of Aedes aegypti and field-collected Daphnia major were exposed to spores added to 50 ml of water in Petri dishes, (c) Plodia interpunctella larvae of various instars were allowed to feed on spore-contaminated artificial diet.…”

Section: Methodsmentioning

confidence: 99%

Unikaryon slaptonleyi sp.nov. (Microspora: Unikaryonidae) isolated from echinostome and strigeid larvae from Lymnaea peregra: observations on its morphology, transmission and pathogenicity

et al. 1983

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SUMMARYA microsporidium, isolated from echinostome and strigeid larval trematodes in Lymnaea peregra, is described as a new species Unikaryon slaptonleyi sp.nov. The nuclei isolated at all stages of development, the disporoblastic sporogony and development in contact with host cell cytoplasm are used to assign the species to the genus Unikaryon. The absence of a vacuolar membrane to isolate the meronts and stages of sporulation from the host cell cytoplasm differentiates this genus from Encephalitozoon. Spores are uninucleate, have 17–21 turns of the polar filament coil and measure 5·0 × 2·8/μm fresh. U. slaptonleyi was isolated from rediae and metacercariae of Echinoparyphium recurvatum and sporo-cysts and cercariae of an unidentified strigeid trematode in L. peregra. It was transmitted in the laboratory to unidentified echinostomes in L. peregra and to unidentified strigeids in Planorbis planorbis by feeding the spores to field-collected snails from which cercariae were already emerging. In these natural and experimental hyperinfections the snail tissues were lightly infected but, in the helminths, much of the parenchyma and germinal tissue was destroyed, so that few cercariae were released and most of those were distorted. Similar heavy infections were produced in Fasciola hepatica in Lymnaea truncatula, when spores were fed to the snails 14 days after miracidial penetration, but even high doses (106 spores/snail) produced only light infections in Schistosoma mansoni in Biomphalaria glabrata, in only 2 out of 9 snails. No infections were obtained in larvae producing xiphidiocercariae in P. planorbis although echinostomes became infected under the same conditions. Of a number of aquatic and terrestrial arthropods tested for susceptibility by feeding or by inoculation of spores into the haemocoele, only Pieris brassicae became infected. In a small proportion of pupae surviving from larvae which had been inoculated with spores at 3rd or 4th instar, there was clear evidence of spore replication.

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The use of a microinjection procedure for large-scale production of the microsporidian Nosema eurytremae in Pieris brassicae

Cited by 14 publications

References 9 publications

Culture of microsporidia from invertebrates in vertebrate cells

Culture of microsporidia from invertebrates in vertebrate cells

An experimental study of the effects of Nosema eurytremae (Microsporida: Nosematidae) on the liver fluke Fasciola hepatica

Unikaryon slaptonleyi sp.nov. (Microspora: Unikaryonidae) isolated from echinostome and strigeid larvae from Lymnaea peregra: observations on its morphology, transmission and pathogenicity

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