Two hundred and seventeen captive great apes (150 chimpanzees, Pan troglodytes; 14 bonobos, Pan paniscus; 53 western gorillas, Gorilla gorilla) and 20 personnel from thirteen European zoos and two African sanctuaries were sampled and examined in order to determine the occurrence of Enterocytozoon bieneusi and species of Encephalitozoon in faecal specimens and to compare the epidemiological situation between zoos and sanctuaries. Microsporidia were detected at all sampling sites. Sequence analyses of ITS amplicons generated by using microsporidia-specific primers determined the presence of microsporidia in 87 samples including 13 humans; since two cases of simultaneous occurrence of Encephalitozoon cuniculi and Enterocytozoon bieneusi were identified, 89 full-length ITS sequences were obtained, namely 78 Encephalitozoon cuniculi genotype I, five E. cuniculi genotype II, two E. hellem 1A and four Enterocytozoon bieneusi. No Encephalitozoon intestinalis-positive samples were identified. This is the first report of Encephalitozoon species and Enterocytozoon bieneusi genotypes in captive great apes kept under various conditions and the first record of natural infection with E. hellem in great apes. A comparison of zoos and sanctuaries showed a significantly higher prevalence of microsporidia in sanctuaries (P<0.001), raising a question about the factors affecting the occurrence of microsporidia in epidemiologically and sanitarily comparable types of facilities.
We investigated the genotypes of Cryptosporidium infecting red squirrels (Sciurus vulgaris L.) in two areas of the Western Alps in Italy. Examination of 141 faecal samples from 70 red squirrels revealed oocysts of Cryptosporidium in 17 animals (24.3%). Based on 18S rRNA gene sequencing, two genotypes of Cryptosporidium species were found: 15 squirrels were positive for the Cryptosporidium ferret genotype and 2 for the Cryptosporidium chipmunk genotype I. The occurrence and intensity of Cryptosporidium infection did not differ between the study areas or sex. More than 85% of the positive animals were adults; however no difference was found between Cryptosporidium infection in the juvenile and adult age groups. Oocysts of the Cryptosporidium ferret genotype measured 5.5 ± 0.3 × 5.2 ± 0.2 µm (shape index 1.06) and the Cryptosporidium chipmunk genotype I 5.8 ± 0.3 × 5.4 ± 0.3 µm (shape index 1.07). Neonatal and adult CD1 and BABL/c mice inoculated with 1 × 103 fresh oocysts of both genotypes did not produce detectable infection.
Parasites of the genus Cryptosporidium Tyzzer, 1910 are one of the most common protistan parasites of vertebrates. Faecal samples from 179 red foxes (Vulpes vulpes [Linnaeus]), 100 grey wolves (Canis lupus Linnaeus), 11 golden jackals (Canis aureus Linnaeus), and 63 brown bears (Ursus arctos Linnaeus) were collected in the Czech Republic, Poland and Slovakia. Samples were examined for the presence of Cryptosporidium spp. using microscopy and PCR/sequence analysis. Phylogenetic analysis based on the small subunit ribosomal RNA (SSU), actin and 60-kDa glycoprotein (gp60) genes using the maximum likelihood method revealed the presence of Cryptosporidium tyzzeri Ren, Zhao, Zhang, Ning, Jian et al., 2012 (n = 1) and C. andersoni Lindsay, Upton, Owens, Morgan, Mead et Blackburn, 2000 (n = 2) in red foxes, C. canis Fayer, Trout, Xiao, Morgan, Lai et Dubey, 2001 (n = 2) and C. ubiquitum Fayer, Santín et Macarisin, 2010 (n = 2) in grey wolves, and C. galli Pavlásek, 1999 in brown bears (n = 1) and red foxes (n = 1). Subtyping of isolates of C. ubiquitum and C. tyzzeri based on sequence analysis of gp60 showed that they belong to the XIId and IXa families, respectively. The presence of specific DNA of C. tyzzeri, C. andersoni and C. galli, which primarily infect the prey of carnivores, is probably the result of their passage through the gastrointestinal tract of the carnivores. Finding C. ubiquitum XIId in wolves may mean broadening the host spectrum of this subtype, but it remains possible this is the result of infected prey passing through the wolf - in this case deer, which is a common host of this parasite. The dog genotype of C. canis was reported for the first time in wolves.