Long‐Term Storage of Infective Microsporidian Spores in Liquid Nitrogen

Maddox, Joseph V.; Solter, Leellen F.

doi:10.1111/j.1550-7408.1996.tb01395.x

Cited by 41 publications

(18 citation statements)

References 8 publications

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“…In contrast, insect-infecting microsporidia are often preserved for extended periods in liquid nitrogen (Maddox and Solter 1996). Future investigations into cell-culture techniques may provide an eective alternative to freezing for long term storage of L. salmonae.…”

Section: Discussionmentioning

confidence: 99%

“…Some species can be stored under refrigeration, in frozen water, lyophilized, or as vacuum-dried quantities and may be viable for greater than 20 years when stored in liquid nitrogen (Maddox and Solter 1996). These species retain viability and have been documented surviving for years at only 4°C in Ringer's solution (Revell 1960;Kramer 1970).…”

Section: Introductionmentioning

confidence: 99%

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Viability of Loma salmonae (Microsporidia) under laboratory conditions

Shaw¹,

Kent

Adamson

2000

Parasitol Res

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The viability of the fish-infecting microsporidian Loma salmonae Morrison and Sprague, 1981 was determined under laboratory conditions by polar filament extrusion and infectivity to chinook salmon (Oncorhynchus tshawytscha). Extrusion rates of isolated spores decreased from 51.0% to 0.0% by 100 days after storage in fresh or sea water at 4 degrees C. Spores stored up to 95 days in either solution infected 80.0-100.0% of exposed chinook, although no spores infected fish at 100 days in one trial. Viability in Earl's balanced salt solution was tested up to 50 days, with 23.7% of spores extruding filaments and 80.0% of exposed chinook becoming infected. Spores frozen to -20 degrees C or -70 degrees C were unable to infect fish.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Viability of Loma salmonae (Microsporidia) under laboratory conditions

Shaw¹,

Kent

Adamson

2000

Parasitol Res

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“…Only few studies report no negative effects of a microsporidian parasite on its native host; although the infection of two lady beetles resulted in prolonged larval development and no higher mortality was recorded (34)(35)(36). We suggest that the prolonged survival time of infected T. minor might increase the possibility of infecting organs other than the gut, such as the ovaries that make vertical transmission of the microsporidian pathogen possible.…”

Section: Discussionmentioning

confidence: 99%

Infection, course of disease and action of Canningia tomici in Tomicus piniperda and Tomicus minor (Coleoptera: Curculionidae)

et al. 2017

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Background and purpose. The pine shoot beetles Tomicus piniperda and Tomicus minor are secondary tree pests attacking weakened Pinus sylvestris and Pinus nigra. Outbreaks occasionally occur, causing considerable economic damage. The microsporidian pathogen Canningia tomici infects T. piniperda as its principal host. Bioassays were used to study the infectivity, vertical transmission, and effects on survival and fecundity of C. tomici on the two beetle species. Austria (Lower Austria, Burgenland and Styria), Finland (Hyytiälä), Poland (Sękocin) (1, 2). T. piniperda occurs throughout Europe, from the northernmost regions of Norway, Finland, and Sweden, to the south of Central Europe. It has spread to China (3) and was accidentally introduced into the USA (4) and Canada (5, 6). The related species T. minor occurs sympatrically with T. piniperda, but uses different stem regions on pine trees. It is widespread throughout Europe, from Scandinavia to the Mediterranean region (2). Depending on the ambient temperature, overwintering adults of both species initiate flight activity in spring, with T. minor flying later than T. piniperda. Following mating, females tunnel mother galleries into the phloem and lay one clutch of eggs. Re-emergence of adult beetles occurs when breeding and oviposition is completed. Subsequently, they bore into shoots of mature pine trees for regeneration feeding, followed by the initiation of a sister brood in uninfested regions of the same tree or in uninfested logs. Both species may infest the same host tree; T. piniperda prefers the lower parts of trees with thick bark for breeding sites, while T. minor prefers trunk with thin bark. Both species bore/tunnel into one-year-old pine shoots for maturation feeding (7,8). Shoot feeding by young callow adults and older adult beetles causes the primary injuries to pine trees, resulting in twig death and weakening of the infested trees (9, 10). However, both beetle species are known as secondary pests in pine stands during the endemic phase. Nonetheless outbreaks occur occasionally, during which the beetles are able to attack vigorous trees (11). This causes considerable economic damage, particularly by T. minor. In addition to infesting living trees, both beetle species infest wind thrown, felled, and firedamaged trees, as well as fresh logs (7,(12)(13)(14). Moreover, these phloeophagous beetles introduce blue stain fungi (e.g. Leptographium wingfieldii, Ophiostoma minus), which interfere with water conduction, accelerate tree death, (15)(16)(17) and reduce the commercial value of timber when it develops blue stain. Materials and methods. Field collected beetles from several locations inBiological control has never been seriously attempted with bark beetles (18,19) and should be reconsidered in light of the results of a study conducted by Kohlmayr et al. (20), which reports the presence of the microsporidium Canningia tomici in cells of the midgut epithelium, adipose tissue, and ovariols of T. piniperda. Little has been reported about the ...

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“…Fresh microsporidian spores used in the experiments were produced in L. dispar (according to Hoch et al 2000), harvested from infected tissues approximately 20 days post inoculation (dpi), and cleaned by Wltration through cellulose tissue and centrifugation. Spore suspensions in distilled water were mixed 1:1 with glycerol and stored in liquid nitrogen to guarantee best spore viability (Maddox and Solter 1996) for no longer than 3 months prior to being used in the experiments.…”

Section: Insects and Pathogensmentioning

confidence: 99%

Effect of midgut infection with the microsporidium Endoreticulatus schubergi on carbohydrate and lipid levels in Lymantria dispar larvae

2009

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Lymantria dispar (Lep., Lymantriidae), an important forest defoliator, is host for a variety of entomopathogenic microsporidia. These are obligate intracellular parasites that completely depend on resources provided by the hosts. The microsporidia infecting L. dispar diVer in the host tissues that become infected as well as in virulence, with presumably diVerent eVects on host physiology. Endoreticulatus schubergi infects the midgut tissue of the host; its virulence is low. We studied the eVects of infection with E. schubergi on carbohydrate and lipid levels in the L. dispar host larva. Trehalose titer in the hemolymph increased with larval age. There were no signiWcant diVerences between infected and uninfected hosts. Glycogen content of the host tissue also increased with age. Levels of this storage carbohydrate were signiWcantly lower in infected larvae at a late stage of infection when insects were either full grown larvae prior to pupation (57.4 § 5.7 and 36.4 § 5.2 g/mg in uninfected controls and infected hosts, respectively) or prepupae (53.2 § 3.4 vs. 34.7 § 4.7 g/mg). Lipid levels in host tissue likewise increased over time and were slightly but not signiWcantly lower in infected L. dispar at a late stage. The results suggest that L. dispar larvae are able to compensate for most of the loss of nurients to the developing microsporidia. DiVerences between these Wndings and the eVects of infection with the virulent microsporidium, Vairimorpha disparis, that infects the host's fat body and causes severe depletion of nutrients in L. dispar larvae are discussed.

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Long‐Term Storage of Infective Microsporidian Spores in Liquid Nitrogen

Cited by 41 publications

References 8 publications

Viability of Loma salmonae (Microsporidia) under laboratory conditions

Viability of Loma salmonae (Microsporidia) under laboratory conditions

Infection, course of disease and action of Canningia tomici in Tomicus piniperda and Tomicus minor (Coleoptera: Curculionidae)

Effect of midgut infection with the microsporidium Endoreticulatus schubergi on carbohydrate and lipid levels in Lymantria dispar larvae

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