FIRST REPORT OF GENUS Cryptosporidium IN CERVIDS SPECIES: Mazama americana, Mazama nana AND Blastocerus dichotomus.

We analyzed Cryptosporidium spp. in fecal samples of wild cervids (Ozotoceros bezoarticus, Blastocerus dichotomus, Mazama nana, Mazama americana, and Mazama bororo) from many Brazilian regions, a fact unprecedented in the literature. Sniffer dogs were used to collect 936 fecal samples of cervids from 14 Brazilian localities. Cervids species were identified using polymerase chain reaction (PCR) performed from genomic DNA extracted from 563 fecal samples of Ozotoceros bezoarticus, Blastocerus dichotomus, Mazama nana, Mazama americana, and Mazama bororo. Cryptosporidium spp. oocyst screening was performed using malachite green negative staining. Nested PCR (nPCR) protocols targeting the 18S rRNA and GP60 genes followed by genetic sequencing were performed for Cryptosporidium spp. detection and Cryptosporidium parvum subtyping, respectively. Nested PCR targeting actin gene and genetic sequencing were performed in samples with non-identified Cryptosporidium species by 18S rRNA amplicon sequencing. The association between the occurrence of Cryptosporidium and the presence of bovines in the same locality was evaluated using Fisher's exact test. The positivity rates of diagnostic methods were compared by McNemar test and the Kappa correlation coefficient. The prevalence rates of Cryptosporidium spp. in cervids were 1.42% (8/563) and 0.36% (2/563) by nPCR and malachite green negative staining, respectively. C. parvum IIaA16G3R1 isolate was identified in three fecal samples from M. americana, two from M. nana and one from B. dichotomus. Cryptosporidium ryanae were found in one sample from B. dichotomus. We identified a new Cryptosporidium genotype, named Cryptosporidium deer genotype BR, from one M. americana fecal sample.

Detection of Cryptosporidium spp. and Giardia duodenalis in small wild mammals in northeastern Brazil.

This study investigated the occurrence of Giardia duodenalis and Cryptosporidium spp. in rodents and marsupials from the Atlantic Forest in southern Bahia, northeastern Brazil. Two hundred and four fecal samples were collected from different forest areas in the municipalities of Ilhéus, Una, Belmonte, and Mascote. Identifications were performed using PCR and nested PCR followed by sequencing of the gdh and tpi genes for G. duodenalis, and the gp60 and Hsp-70 genes for Cryptosporidium. The total frequency of positive PCR samples for both G. duodenalis and Cryptosporidium spp. was 5.4% (11/204). Giardia duodenalis occurred in 2.94% (4/136) of rodents and 2.94% (2/68) of marsupials. The prevalence of Cryptosporidium in rodents and marsupials was 1.47% (2/136) and 4.41% (3/68), respectively. In the areas sampled, the frequency of parasitism was 50% (7/14), while the Mascote region alone had no parasitized animals. The G. duodenalis subgenotype AI was identified in the rodent species Hylaeamys laticeps, Oecomys catherinae, Oligoryzomys nigripes and Akodon cursor, and in the marsupials Gracilinanus agilis and Monodelphis americana. In the rodents Rhipidomys mastacalis, H. laticeps and in the marsupial Marmosa murina the protozoa Cryptosporidium fayeri, Cryptosporidium parvum and Cryptosporidium ubiquitum with subtypes IIa and IVg by the gp60 gene were found. In conclusion, this study provides the genetic characterization of Giardia and Cryptosporidium species and genotypes in rodents and marsupials. And, these findings reinforce that the rodent and marsupial species mentioned above play a role as new hosts for Giardia and Cryptosporidium.