Cryptosporidiosis is a parasitic disease responsible for gastroenteritis in reptiles. The main species affecting snakes are C. serpentis and C. varanii, although recent molecular studies have identified other species and genotypes. Snakes can act as asymptomatic carriers excreting intermittently low numbers of oocysts in feces impossible to detect using microscopic routine techniques. The main goal of this research was to provide information on the prevalence of Cryptosporidium species and genotypes in captive snakes from Italy. For this purpose, a total of 21 fecal samples were collected from animals with no apparent clinical symptoms of cryptosporidiosis at the time of sampling. Fecal samples were diluted in PBS, filtered through a sieve and centrifuged. 20 ml of the sediment were examined for the presence of Cryptosporidium oocysts by a direct immunofluorescence antibody test. Cryptosporidium species were determined by nested PCR of a SSU rRNA gene fragment and RFLP analysis with the endonucleases Sspl and Vspl. Cryptosporidium oocysts were observed in 15 and 21 sample (71%) using IFAT, whereas PCR products of the SSU rRNA locus were obtained from 14 snakes (67%). Twelve samples tested positive to both techniques. PCR-RFLP analysis of SSU rRNA revealed banding patterns distinctive for the Cryptosporidium mouse genotype (10/14) and C. muris (3/14). The low intensity of the remaining sample prevented its accurate identification by RFLP. C. muris and the Cryptosporidium mouse genotype have previously been reported in captive snake, although they had been not considered true parasites of these animals, result only from the ingestion of infected rodents. As cryptosporidiosis is often asymptomatic in snakes and oocysts of Cryptosporidium species affecting them are difficult to differentiate from those just passing through the gastrointestinal tract, application of molecular techniques are needed to achieve a correct identification of the species or genotypes involved.
Molecular identification of Cryptosporidium isolates from captive snakes in Italy / Diaz Fernandez P.D.; Rota S.; Marchesi B.; Soilan Lopez M.; Lopez sandez C.M.; Panadero Fontan R.; Dez Banos P.; Morrondo Pelayo P.; Poglayen G.. - STAMPA. - XII:(2011), pp. 241--. (Intervento presentato al convegno XII Congreso iberico Parasitologia tenutosi a Zaragoza nel 5-8 luglio 2011).
Molecular identification of Cryptosporidium isolates from captive snakes in Italy
MARCHESI, BARBARA;POGLAYEN, GIOVANNI
2011
Abstract
Cryptosporidiosis is a parasitic disease responsible for gastroenteritis in reptiles. The main species affecting snakes are C. serpentis and C. varanii, although recent molecular studies have identified other species and genotypes. Snakes can act as asymptomatic carriers excreting intermittently low numbers of oocysts in feces impossible to detect using microscopic routine techniques. The main goal of this research was to provide information on the prevalence of Cryptosporidium species and genotypes in captive snakes from Italy. For this purpose, a total of 21 fecal samples were collected from animals with no apparent clinical symptoms of cryptosporidiosis at the time of sampling. Fecal samples were diluted in PBS, filtered through a sieve and centrifuged. 20 ml of the sediment were examined for the presence of Cryptosporidium oocysts by a direct immunofluorescence antibody test. Cryptosporidium species were determined by nested PCR of a SSU rRNA gene fragment and RFLP analysis with the endonucleases Sspl and Vspl. Cryptosporidium oocysts were observed in 15 and 21 sample (71%) using IFAT, whereas PCR products of the SSU rRNA locus were obtained from 14 snakes (67%). Twelve samples tested positive to both techniques. PCR-RFLP analysis of SSU rRNA revealed banding patterns distinctive for the Cryptosporidium mouse genotype (10/14) and C. muris (3/14). The low intensity of the remaining sample prevented its accurate identification by RFLP. C. muris and the Cryptosporidium mouse genotype have previously been reported in captive snake, although they had been not considered true parasites of these animals, result only from the ingestion of infected rodents. As cryptosporidiosis is often asymptomatic in snakes and oocysts of Cryptosporidium species affecting them are difficult to differentiate from those just passing through the gastrointestinal tract, application of molecular techniques are needed to achieve a correct identification of the species or genotypes involved.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
