Occurrence and molecular characterization of Cryptosporidium sp. in sheep

Abstract

Considered a zoonosis of utmost importance, cryptosporidiosis has a worldwide distribution and can infect mammals, birds, reptiles, and amphibians. It is caused by a highly resistant protozoan present in the environment and can cause death in immunosuppressed individuals and pups, as well as in farm animals such as cattle and sheep, generating losses. The aim of this study was to investigate the presence of Cryptosporidium spp. in sheep feces from the farms of Western Paraná, which have different management styles, and compare the results with their respective management methods. One hundred and forty-four stool samples were collected (69 from Property 1 and 75 from Property 2) and analyzed using a fecal smear on slides after staining by the modified Ziehl-Neelsen method. Samples tested positive by this method were subjected to nested PCR and the products obtained were sent for sequencing to determine the species. While 82.60% of the samples from Property 1 were tested positive, only 36% of the samples from Property 2 were tested positive. On analyzing the sequencing data, it was observed that the Cryptosporidium species of samples from Property 1 showed high similarity to Cryptosporidium xiaoi and those from Property 2, to Cryptosporidium ubiquitum. The reason for divergence in results can be attributed to differences in management systems adopted by each property, thus showing the importance of detecting carrier animals, as they can contaminate the environment, especially the water sources, and spread the disease to humans and other animals. Key words: Cryptosporidium ubiquitum. Cryptosporidium xiaoi. Nested-PCR. Sheep. Ziehl-Neelsen. ResumoConsiderada uma zoonose de extrema importância, a cryptosporidiose possui distribuição mundial e pode infectar mamíferos, aves, répteis e anfíbios. É causada por um protozoário muito resistente no ambiente, podendo gerar mortes em indivíduos imunossuprimidos e filhotes, principalmente em animais de produção como bovinos e ovinos, gerando prejuízos. O objetivo desse trabalho foi pesquisar a presença de Cryptosporidium spp. em fezes de ovinos de propriedades rurais do Oeste do Paraná, que possuem manejos diferenciados e comparar os resultados encontrados com o respectivo manejo. Foram coletadas 144 amostras de fezes, sendo 69 da Propriedade 1 e 75 da Propriedade 2, analisadas pelo esfregaço de fezes em lâminas após a coloração pelo método de Ziehl-Neelsen modificado. As amostras positivas por esse método foram submetidas à Nested-PCR sendo os produtos obtidos enviados ao sequenciamento para a determinação da espécie. A Propriedade 1 apresentou positividade em 82,60% das amostras, enquanto a Propriedade 2 obteve 36% de amostras positivas. Foi encontrada alta similaridade para Cryptosporidium xiaoi na primeira propriedade e Cryptosporidium ubiquitum na segunda propriedade. A divergência nos resultados pode ser comprovada pela diferente forma de manejo adotada em cada propriedade, mostrando a importância de detectar os animais portadores, uma vez que estes podem contami...

Cryptosporidium in Dairy Cattle, Sheep, Humans, and Manure in and around Adama and Asella Towns, Oromia Regional State, Ethiopia

“…However, the current nding is lower than the reports from Mexico (Romero-salas et al, 2016), Brazil (Snak et al, 2017) and Iraq (Al-zubaidi, 2017) with high prevalence; 72.5%, 59%, and 48%, respectively.…”

“…These discrepancies in prevalence might be attributed to the management system in which the present study collected the sample from extensively reared sheep. The sampling animals (sheep) in this study are kept out of doors and are less exposed to Cryptosporidium oocyst than intensively managed sheep, whose crowding and con nement to small areas favor environmental contamination with oocyst and exposure to infection (Snak et al, 2017). Furthermore, in an extensive management system, the oocyst excreted from the oocyst spreads to a large surface area, resulting in low infection pressure (Geurden et al, 2006).…”

Cryptosporidium occurrence in ruminants from the North Pioneer mesoregion of Paraná, Brazil

“…In the same city, 12.4% of the calves were found to be positive using this same technique in a similar number of samples (FEITOSA et al, 2004). In the State of Paraná, recent research in the Western region showed high positivity in adult sheep (59%), however the authors evaluated few lambs (7/144) (SNAK et al, 2017) The infection rate among lambs was 43% higher than that seen in sheep. Although this difference was not significant, these results were similar to those published by Causapé et al (2002) and Paz & Silva et al (2014).…”

“…In Brazil, studies on cryptosporidiosis in ruminants have primarily been performed in the Southeast, mainly in the states of Rio de Janeiro (COSENDEY et al, 2008;FIUZA et al, 2011), Minas Gerais (GARCIA & LIMA, 1993;GARCIA & LIMA, 1994;LIMA et al, 2013), andSão Paulo (AQUINO et al, 2015;COELHO et al, 2016;FEITOSA et al, 2004;FÉRES et al, 2009;PAZ E SILVA et al, 2013SEVÁ et al, 2010;SILVA-JUNIOR et al, 2011). In the state of Paraná, infection with Cryptosporidium was evaluated in captive birds (NAKAMURA et al, 2009), dogs with diarrhea (NAVARRO et al, 1997), horses (FUJII et al, 2014, wild birds and mammals (SNAK et al, 2015), cattle (TOLEDO et al, 2017 and sheep (SNAK et al, 2017). However, epidemiological studies have not been conducted on cryptosporidiosis in ruminants of the north pioneer mesoregion of Paraná.…”

“…Techniques such as the modified Ziehl-Neelsen method, direct fluorescent antibody test, negative staining method or ELISA, with or without subsequent PCR, are widely used in the study of cryptosporidosis in ruminants. Among these techniques, PCR has the highest sensitivity (MORGAN et al, 1998;REKHA et al, 2016;TAHVILDAR-BIDEROUNI & SALEHI, 2014); moreover, the reported rates of occurrence of Cryptosporidium vary between studies (CAUSAPÉ et al, 2002;EDERLI, et al, 2004;FÉRES et al, 2009;PAZ & SILVA et al, 2014;SNAK et al, 2017;TOLEDO et al, 2017). Higher infection rates are reported with the use of a stool oocyst-concentration technique, such as Sheather's flotation method (COKLIN et al, 2007;COUTO et al, 2014), probably due to its high sensitivity (REKHA et al, 2016).…”