Growth of Nosema algerae in Pig Kidney Cell Cultures*

SYNOPSIS. Nosema algerae, a microsporidan parasite of mosquitoes, can infect pig kidney cell cultures. Spores germinated in the culture medium, infected the cells within 30 min of germination, multiplied, and produced spores. The early developmental stages in the N. algerae life cycle are described.     

Ultrastructure of the Peripheral Zone of a Glugea-Induced Xenoma*

SYNOPSIS. The Glugea stephani-induced xenoma in the winter flounder, Pseudopleuronectes americanus, is a large spherical host-parasite complex, up to 4.0 mm in diameter, with the host and parasite components of the xenoma being most active in the peripheral zone. The xenoma has an extensive periodic acid-silver methenamine-positive surface coat covering the plasma membrane. The surface of this membrane is amplified by the presence of numerous folds and fine tubular extensions. The peripheral zone of the xenoma contains many host-cell mitochondria in addition to numerous microsporidan parasites. At the ultrastructural level, the peripheral zone of the host-cell cytoplasm appears normal. Inside the peripheral region of the 0.4–1.0 mm xenoma, the host-cell component largely disintegrates in the presence of microsporidan parasites undergoing sporogenesis.     

Observations on the Spores of Pleistophora gigantea (Thélohan, 1895) Swellengrebel, 1911, a Microsporidan Parasite of the Fish Crenilabrus melops*

SYNOPSIS. After 1914 protozoologists have generally agreed that Pleistophora gigantea (Thélohan, 1895) Swellengrebel, 1911, Ichthyosporidium giganteum (Thélohan, 1895) Swarczewsky, 1914, and I. phymogenes Caullery and Mesnil, 1905, are identical. Because no polar filament was found in the spores, however, some authors have followed Swarczewsky in considering this species to be a haplosporidan, while others have persisted in thinking it a microsporidan. Using preserved material that Swellengrebel saved from a tumor on which he based his studies, we have found a polar filament in the spores both with the PAS reaction and with the electron microscpe. This new information removes the only basis for the doubt which some authors have entertained, that Thélohan and Sweliengrebel correctly considered the parasite to belong to the Microsporida. Since Pleistophora gigantea is believed to be identical with I. phymogenes, recently selected by Sprague as type species of genus Ichthyosporidium Caullery and Mesnil, 1905, then Ichthyosporidium, originally assigned to the Haplosporida, must be regarded as a microsporidan genus. Whether it is distinct from all other microsporidan genera is a matter needing further consideration.     

Laboratory and Field Studies on Glugea stephani (Hagenmuller), a Microsporidan Parasite of Pleuronectid Flatfishes*

SYNOPSIS. The microsporidan Glugea stephani is a common parasite of juvenile English sole (Parophrys vetulus) in Yaquina Bay, Oregon. Field observations indicated that fish became infected only in the upper estuary where summer temperatures were above 15 C and the incidence of infection reached 79.8% in the late fall. Laboratory infections developed and parasite growth occurred only at or above 15 C. The parasite was successfully transmitted to juvenile English sole by brine shrimp (Artemia salina) and amphipod (Corophium spinicorne) vectors as well as by direct ingestion of spores by the host. Infections that resulted from ingestion of spore-carrying vectors were much heavier than those resulting from the direct ingestion of spores. The speckled sanddab (Citharichthys stigmaeus), a nonpleuronectid flatfish, and chum salmon (Oncorhynchus keta) were refractory to G. stephani infection in the laboratory. Results of this study suggest that G. stephani is potentially lethal to young pleuronectid flatfishes when heavy infections involve the entire intestine and reduce the capacity to absorb nutrients. Under these circumstances, starvation is probably the direct or indirect cause of death. The restriction of infection to fish that reach the upper estuary very likely mitigates the impact of G. stephani caused mortality on the entire English sole population on the Yaquina Bay nursery ground.     

Nosema algerae: Infection of the white mouse by a mosquito parasite

Spores of Nosema algerae, a microsporidan parasite of mosquitoes, were subcutaneously injected into the ears, tail, and feet of white mice. Infections were transient and localized at the injection sites. Spore germination tests in blood plasma indicated that it is unlikely that spores injected by an infected mosquito bite would result in an infection.     

In vivo propagation of a microsporidan pathogenic to insects

Equivalent numbers of spores were produced when the microsporidan Nosema necatrix was propagated in either Trichoplusia ni or Heliothis zea. Maximum spore production was obtained at an inoculum level of 1 × 10⁵ spores/ml. Larvae inoculated 5 days post-hatching contained 1.6 × 10⁹ spores/gram larva after an incubation period of 21 days. Temperature optima for the parasite are 21–26°C in both hosts.     

Spore Dimorphism in a Microsporidan Isolate*

A microsporidan isolate currently considered to represent a mixed infection of Nosema necatrix Kramer, 1965 and Thelohania diazoma Kramer, 1965 was subjected to cultivation in hosts held at various temperatures. The ratio of the Nosema (monospore) to the Thelohania (octospore) forms at these temperatures was found to vary from 1:1 at 16 C to 1:0 at 32 C. Isolation technics using mechanical, temperature and temporal methods separated monosporous from octosporous forms for inoculation purposes. However, microscopic examination of hosts receiving these inocula revealed the presence of both monospores and octospores. Electrophoretic analysis of monospores and monospore-octospore mixtures indicated equivalent hydrophobic protein spectra. These observations suggest that this isolate has the ability to produce either single spores or spores in groups of eight. This microsporidan was not considered a member of the genus Stempellia since spores in groups of 2 or 4 were not observed. Retention of the name Nosema necatrix Kramer is suggested.     

Ecology of freshwater snails in south-western Nigeria. I: Distribution and habitat preferences

Dry and rainy season investigations of diverse freshwater habitats in south-western Nigeria revealed fourteen species of snail comprised of nine pulmonates: Biomphalaria pfeifferi Krauss, Bulinus globosus Morelet, Bulinus rohlfsi Clessin, Lymnaea natalensis Krauss, Physa ( Aplexa) waterloti Germain, Bulinus forskali Ehrenberg, Gyraulus costulatus Krauss, Ferrissia sp, Segmentorbis sp. and five prosobranchs namely, Lanistes libycus Morelet, Lanistes ovum Peters, Pila wernei Philippi, Potadoma moerchi Reeve and Melanoides tuberculata Müller.The influence of diverse chemical and physical properties of water on their occurrence as well as interspecific and snail-plant relationships are discussed and maps showing the distribution of the species encountered are presented. Among the intermediate hosts of Schistosoma, B. globosus was widely distributed and common, B. pfeieri was also widely distributed but infrequent and B. rohlfsi was rare.     

Infestation by the mite Harpirhynchus nidulans in the Bearded Tit Panurus biarmicus

Capsule Mites sometimes induced voluminous subcutaneous cysts in featherless parts.

Aims To describe the first reported infestation by the skin-dweller mite Harpirhynchus nidulans in Bearded Tits Panurus biarmicus and for the Timaliidae family, to detect possible fitness costs for the host and to determine the distribution of the parasite within the distribution range of the host.

Methods Parasites were identified using a microscope. Wing-length and body mass were recorded on both uninfected and infected birds captured at different times during the year. We also considered historical data, and contact was made with 32 European ringing stations to identify the distribution range of the parasite.

Results Subcutaneous reproduction of the mite Harpirhynchus nidulans induced the development of voluminous dermal nodules in Panurus biarmicus. There were no differences in body mass or wing-length with respect to infestation. In the south of France, prevalence changed from 10.6% in spring to 4.7% in autumn. Both sexes are equally parasitized. Occurrence of dermal cysts is reported from several southern European populations of Bearded Tits, whereas it seems to be absent from northern latitudes.

Conclusion The occurrence of a Harpirhynchus mite in wild bird populations is reported for the first time. We consider aspects of its biology, host–parasite system, host-specificity, co-adaptation of the mite reproductive cycle to the social dynamics of its host and metapopulational host–parasite dynamics.     

Effects of host condition on susceptibility to infection, parasite developmental rate, and parasite transmission in a snail-trematode interaction

Whether or not organisms become infected by parasites is likely to be a complex interplay between host and parasite genotypes, as well as the physiological condition of both species. Details of this interplay are very important because physiology‐driven susceptibility has the potential to confound genetic coevolutionary responses. Here we concentrate on how physiological aspects of infection may interfere with genetic‐based infectivity in a snail–trematode (Potamopyrgus antipodarum/Microphallus sp.) interaction by asking: (1) how does host condition affect susceptibility to infection? and (2) how does host condition affect the survival of infected individuals? We manipulated host condition by experimentally varying resources. Contrary to our expectation, host condition did not affect susceptibility to infection, suggesting that genetics are more important than physiology in this regard. However, hosts in poor condition had higher parasite‐induced mortality than hosts in good condition. Taken together, these results suggest that coevolutionary interactions with parasites may depend on host condition, not by altering susceptibility, but rather by affecting the likelihood of parasite transmission.     

Evaluation of naphthoquinones identified the acetylated isolapachol as a potent and selective antiplasmodium agent

Abstract

This study reports on the design, synthesis and antiparasitic activity of three new semi-synthetic naphthoquinones structurally related to the naturally-occurring lapachol and lapachone. Of the compounds tested, 3-(3-methylbut-1-en-1-yl)-1,4-dioxo-1,4-dihydronaphthalen-2-yl acetate (1) was the most active against Plasmodium falciparum among both natural and semi-synthetic naphthoquinones, showing potent and selective activity. Compound 1 was able to reduce the in vitro parasite burden, in vitro parasite cell cycle, as well as the blood parasitemia in Plasmodium berghei-infected mice. More importantly, infection reduction under compound 1-treatment was achieved without exhibiting mouse genotoxicity. Regarding the molecular mechanism of action, this compound inhibited the hemozoin crystal formation in P. falciparum treated cells, and this was further confirmed by observing that it inhibits the β-hematin polymerization process similarly to chloroquine. Interestingly, this compound did not affect either mitochondria structure or cause DNA fragmentation in parasite treated cells. In conclusion, we identified a semi-synthetic antimalarial naphthoquinone closely related to isolapachol, which had stronger antimalarial activity than lapachol.     

A new genus of Diplommatinidae from the Andaman Islands (Gastropoda: Caenogastropoda: Cyclophoroidea)

ABSTRACT

Examination of the syntypes of Pupa (Pupisoma) constrictus Godwin-Austen, 1895 revealed that this species does not belong to the eupulmonate Pupilloidea, but to the caenogastropod Diplommatinidae. Based on conchological characters it could not be placed to any of the existing diplommatinid genera, therefore a new genus, Moderata n. gen. is erected, which is characterised by a conical shell, a discontinuous aperture and most importantly, a conspicuous constriction half a whorl behind the aperture. Pupisoma bilamellatum Benthem Jutting, 1958 and P. unilamellatum Benthem Jutting, 1958 are moved to the assimineid genus Acmella W. T. Blanford, 1869.     

Type materials of the species of the Planorbidae (Mollusca,Gastropoda, Hygrophila) described by Shuichi Mori

ABSTRACT

The planorbid species that were described by Shuichi (Syuiti) Mori from Japan have long been enigmatic and their taxonomy has been highly confused. This situation is also an impediment to action for the conservation of Japanese planorbids which are one of the most threatened freshwater snail groups in the country. Based on our examination of Shuichi Mori’s collection stored in the Kyoto University Museum, type materials of the taxa described by him are re-illustrated and re-examined for the first time. Our examination suggested that Gyraulus amplificatus (Mori, 1938) is not a morphological form of Gyraulus biwaensis but a distinct species. Gyraulus pulcher (Mori, 1938) can be regarded as a junior synonym of Gyraulus chinensis.     

The rate of development of a microsporidan in moth cell culture.

A microsporidan parasite of the forest tent caterpillar Malacosoma disstria infected cells and replicated in vitro in a line from the moth Heliothis zea. After spore germination, the incidence of infected cells increased with time until leveling off with sporulation. During the first 24 hr, there was a static number of parasites, followed by a 2-day logarithmic growth phase during which the population doubled five to six times. The growth rate was 9 to 11 hr per population doubling. Sporulation commenced on day 3, and 40 to 50 spores were recovered from each infected cell. The life cycle was completed within 6 days, culminating in spores that were infectious for cultured cells. The antibiotic fumagillin at a dose of 1 ppm in the culture medium was microsporida-static.     

Evolutionary aspects of the metapopulation dynamics of Biomphalaria pfeifferi,the intermediate host of Schistosoma mansoni

Combining genetic and demographic data is a powerful approach to study adaptation process and evolutionary forces acting in natural populations. We focus on the freshwater snail Biomphalaria pfeifferi, the intermediate host of Schistosoma mansoni. Twenty‐one populations sampled in the south of Madagascar were genotyped at six microsatellite loci. Demographic parameters and parasitic prevalence were estimated monthly over the year preceding the genetic sampling. Our results indicate that populations experience recurrent bottlenecks and size fluctuations, which strongly depresses the genetic diversity within population. The recolonization of depleted sites involves genetically differentiated immigrants. We detected frequent migration events along rivers and rare migration events between watersheds. This explains the high level of differentiation observed among populations. The negative regression observed between the prevalence of S. mansoni and the genetic diversity of B. pfeifferi populations indicates that host consanguinity may affect prevalence through the genetic mechanisms involved in resistance. Coevolutionary outcomes are also influenced by the relative migration rates of snails and flukes, but the parasite local adaptation may be prevented by rare long distance dispersal in snails and the phylogeographical patterns of colonization of both hosts and snails.     

Antileishmanial activities and mechanisms of action of indole-based azoles

Two 3-(α-azolylbenzyl)indoles were evaluated against Leishmania amastigotes. Both compounds proved to be very active against intracellular and axenic amastigotes. The IC₅₀ values of the imidazole derivative, PM17, and the triazole analogue, PM19, against L. mexicana axenic amastigotes, were 4.4 ± 0.1 and 6.4 ± 0.1 μM, respectively. Against intracellular amastigotes, PM17 produced a 66% decrease of leishmanial burden at 1 μM and PM19 had an IC₅₀ of 1.3 μM. In a Balb/c mice model of L. major leishmaniasis, administration of PM17 led to a clear-cut parasite burden reduction: 98.9% in the spleen, 79.0% in the liver and 49.9% in the popliteal node draining the cutaneous lesion. As anticipated, it was brought to the fore that PM17 decreases ergosterol biosynthesis leading to membrane fungal cell alterations. Moreover it was proved that this imidazole antifungal agent induces a parasite burden-correlated decrease in interleukine-4 production both in the splenocyte and the popliteal node of the mouse.     

Relationship between prevalence of trematode parasite <Emphasis Type="Italic">Diplostomum</Emphasis> sp. and population density of its snail host <Emphasis Type="Italic">Lymnaea stagnalis</Emphasis> in lakes and ponds in Finland

Diplostomum sp. is a trematode parasite that infects aquatic snails, e.g. Lymnaea stagnalis (Gastropoda: Lymnaeidae), fish, and fish-eating birds. Ponds and lakes (n = 28) located in Finland between latitudes 61°45′N and 65°30′N were sampled for L. stagnalis, the first intermediate host for Diplostomum sp. L. stagnalis were found in 22 sites out of 28, and Diplostomum sp. in 10 of the 22 snail populations. Among the L. stagnalis populations that were infected by Diplostomum sp., the mean prevalence was 12.8%. Diplostomum sp. occurred in only one out of the seven L. stagnalis populations in four large lakes, but in 9 out of the 16 L. stagnalis populations in small lakes. In the pooled data, a positive correlation (r _s = 0.427; P = 0.047; n = 22) between L. stagnalis density and Diplostomum sp. prevalence was found. The results suggest that Diplostomum sp. is fairly common in L. stagnalis populations in small lakes but rare in large lakes. Furthermore, although trematode parasites, in general, should have a negative effect on snail population density, the study indicates that the relationship between host density and parasite prevalence may greatly differ for individual trematode species, such as Diplostomum sp.     

Evaluating shell variation across different populations of a freshwater snail

The freshwater snail Potamopyrgus antipodarum has become a model system for studying invasion ecology, host–parasite coevolution, the maintenance of sexual reproduction and ecotoxicology. One understudied aspect of this snail is the variation in morphology within and among populations, which could provide insights into ecological differences across its native range. In this study of 17 New Zealand lake populations of P. antipodarum we used linear measurements and geometric morphometrics to compare several aspects of shell size and shape. We found that, except for those from Lake Te Anau, most lake populations of P. antipodarum differ slightly in shape, but differ significantly in size and in the presence of spines, where larger and spinier snails are found in deeper regions. These striking distinctions in size and shell armature, but not in shape, suggest that the various components of form are under different selective regimes. Snails from Lake Te Anau are different in both shape and size, implying that this population is diverging from the rest of the species in multiple ways, making it an interesting study population for further research.     

Sulla penetrazione di Penicillium digitatum Sacc. e Penicillium italicum Wehmer nei frutti degli agrumi

Summary

The A., with a series of controls and investigations, accomplished on different groups of Oranges and following various forms of artificial infection with Penicillium digitatum and Penicillium italicum, renders evident the different reactions that these moulds have with regard to the resistance of the cuticle and the epidermic and under-epidermic stratums of the peel of the controlled fruits. Moreover the A. confirms that both the P. digitatum and the P. italicum can act as «parasite of injury» and as «parasite of contact».