Cryptosporidium fragile sp. n. (Apicomplexa) is described from black-spined toads, Duttaphrynus melanostictus (Schneider) (Amphibia, Anura, Bufonidae) from the Malay Peninsula. The parasitized animals were directly imported from Malaysia and harboured C. fragile at the time of arrival. Oocysts were subspherical to elliptical with irregular contour in optical section, measuring 6.2 (5.5-7.0) × 5.5 (5.0-6.5) µm. Oocyst wall was smooth and colourless in light microscopy. The endogenous development of C. fragile in the stomach of black-spined toad was analysed in detail using light and electron microscopy. Cryptosporidian developmental stages were confined to the surface of gastric epithelial cells. In transmission experiments, C. fragile has not been infective for one fish species, four amphibian species, one species of reptile and SCID mice. Full length small subunit rRNA gene sequence was obtained. Phylogenetic reconstruction revealed distinct status of C. fragile within the clade of species with gastric localisation including Cryptosporidium muris Tyzzer, 1907, Cryptosporidium serpentis Levine, 1980 and Cryptosporidium andersoni Lindsay, Upton, Owens, Morgan, Mead et Blagburn, 2000. Described characteristics differentiate C. fragile from the currently recognized Cryptosporidium species. Our experience with the description of C. fragile has led us to revise the recommended criteria for an introduction of a new Cryptosporidium species name. C. fragile is the first species described and named from an amphibian host. Its prevalence of 83% (15/18) in black-spined toads within the 3 months after importation calls for strict quarantine measures and import regulation for lower vertebrates.
Anaerobic gut fungi of the class Neocallimastigomycetes are of great importance for herbivorous animals. Their immediate colonization and mechanical breakdown of plant particles pave the way for highly efficient enzymatic fermentation of complex plant polysaccharides. Neocallimastigomycetes are found in a variety of herbivores, yet so far studies almost exclusively investigated domestic or captive animals. Here, the occurrence and diversity of Neocallimastigomycetes in two different populations of sympatric, wild African forest elephants and forest buffaloes were determined. In both hosts together, a total of 16 species-equivalent Operational Taxonomic Units (OTUs) (0.05 cut-off level) were generated. Buffaloes harboured four and elephants five anaerobic fungi genera or genus-equivalent taxa, respectively, with four genera occurring in both hosts. In elephants the majority of gut fungi group within a cluster of yet unknown Neocallimastigomycetes. Similarly, some anaerobic fungi found in buffaloes form a genus-equivalent cluster with likewise undescribed gut fungi. Sequences grouping in these two clusters could potentially qualify as representatives of new anaerobic fungi genera. Further, three sequences have not yet been encountered in any study and cannot be assigned to any genus or genus-equivalent Neocallimastigomycetes taxon. Whether these sequences also represent putative new lineages needs further investigation.
Two experimental trials were performed to elucidate the role of rodents in the life cycle of Hepatozoon species using snakes as intermediate hosts. In one trial, two ball pythons, Python regius Shaw, 1802 were force fed livers of laboratory mice previously inoculated with sporocysts of Hepatozoon ayorgbor Sloboda, Kamler, Bulantová, Votýpka et Modrý, 2007. Transmission was successful in these experimentally infected snakes as evidenced by the appearance of intraerythrocytic gamonts, which persisted until the end of trial, 12 months after inoculation. Developmental stages of haemogregarines were not observed in histological sections from mice. In another experimental trial, a presence of haemogregarine DNA in mice inoculated with H. ayorgbor was demonstrated by PCR in the liver, lungs and spleen.
Pterygosoma livingstonei sp. n. collected from the Kenyan lizard Agama caudospinosa Meek shows morphological affinities with other South African congener species parasitizing lizards of the genus Agama, especially with P. triangulare Lawrence, 1936, but it differs in having glabrous genua II and III. P. livingstonei shows affinities with the Lawrence's hispida species group in the characters of genital and peripheral setae. This new species was found concentrated in a nuchal ''mite pocket-like structure'', a behaviour previously unreported among species belonging to the genus Pterygosoma. Mite pockets (or acarodomatia, acarinaria) of lizards typically house damaging chigger mites, and are usually interpreted as the evolutionary host's response to limit damage caused by parasites. Because scale mites are permanent ectoparasites and less damaging than seasonally occurring larval trombiculids, the heavy infestation by P. livingstonei in the nuchal skin folds of its host is interpreted as a consequence of the best utilisation of an available protected site by these mites that spend their entire life cycle on their host and whose primitive body shape prevents them from seeking shelter beneath the scales of their lizard host.
Coprological examination of New Caledonian geckoes of the genus Rhacodactylus Fitzinger, 1843 revealed two new species of coccidia. Isospora leachiani sp. n. from R. leachianus (Cuvier, 1829) has oval, colourless oocysts, measuring 21-26 × 16-18.5 µm. Sporocysts are ellipsoidal, 11-12.5 × 6.5-8 µm, with distinct Stieda and substieda bodies. Oocysts of Isospora sykorai sp. n. from R. ciliatus (Guichenot, 1866) are elongately oval to cylindrical, 20-23.5 × 11-14 µm; sporocysts of this species are ellipsoidal, 10-11.5 × 7-8 µm, with a slightly pointed end and Stieda and substieda bodies. Infected geckoes did not exhibit any alteration of their health status.
During a survey of the coccidian parasites of reptiles, caryosporan oocysts were found in the faeces of wild and captive European viperid snakes Vipera berus (L.) and V. ammodytes (L.). Thirty two of 37 examined V. herus (86%) and 9 of 17 examined V. ammodytes (53%) specimens were found to be passing caryosporan oocysts. Morphological characters of all caryosporan isolates were identical and fitted well with the description of Caryospora simplex Léger, 1904. Experimental inoculation of severe combined immunodeficient (SC1D) mice with seven isolates of C. simplex from V. berus or V. ammodytes confirmed the heteroxenous life cycle pattern, for the first time for isolates of evidently European origin. Caryosporan developmental stages were observed in the connective tissues of the nose, cheeks, ear and scrotum in all inoculated SCID mice. V. berus and V. ammodytes represent new hosts for C. simplex. The present paper represents the first widely based report on coccidian parasites of the genus Caryospora Léger in European viperids. Our findings indicate a wide distribution of C. simplex throughout the range of distribution of snakes of the genus Vipera.