DETECÇÃO DE INFECÇÕES POR Cryptosporidium spp. EM FILHOTES CANINOS POR MÉTODOS MOLECULARES / DETECTION OF Cryptosporidium spp. INFECTIONS IN PUPPIES BY MOLECULAR METHODS

Autor: Marcelo Vasconcelos Meireles, Luiz da Silveira Neto, S. V. Inácio, Katia Denise Saraiva Bresciani, Mariele Fernanda da Cruz Panegossi, A. E. G. Watanabe
Rok vydání: 2013
Předmět:
Zdroj: Ars Veterinaria. 29:50
ISSN: 2175-0106
0102-6380
DOI: 10.15361/2175-0106.2013v29n4p50
Popis: O protozoario Cryptosporidium spp. e considerado um importante patogeno em termos de saude publica (CIELOSZYK et al., 2012), sendo principalmente associado a baixo poder socioeconomico e precarias condicoes de saneamento basico (ASSIS et al., 2013). Caes eliminam oocistos fecais, com ou sem diarreia, sendo considerados potenciais fontes de infeccao humana (WANG et al., 2012), apesar de isto nao ter sido confirmado experimentalmente (BOWMAN & LUCIO-FORSTER, 2010; UEHLINGER et al., 2013). Detectar infeccoes por Cryptosporidium spp. em amostras fecais de filhotes caninos por meio da reacao em Cadeia da Polimerase-Nested (Nested-PCR), foi o objetivo deste estudo. As amostras, previamente identificadas e congeladas, foram destinadas a inquerito molecular. A extracao de DNA foi realizada por meio do kit comercial QIAamp DNA Stool (Qiagen) e a tecnica de nested-PCR foi empregada para a amplificacao de fragmentos do gene da subunidade 18S do RNA ribossomico. Um total de 200 caes foram examinados, sendo 100 machos e 100 femeas, 111 de padrao racial determinado e 89 sem raca definida (SRD). Destes, 81 animais, 43, 48 e 28 tinham ate dois, de dois a tres; de tres a seis e de seis a doze meses, respectivamente. Conforme sua origem, os animais eram provenientes dos Municipios de Aracatuba e Votuporanga, SP, sendo que 126 eram de domicilios; 11 mantidos em centros de Zoonoses; 50 de Pet Shops; 12 de um criatorio e uma (0,5%) era errante e havia sido adotada. A prevalencia amostral de Cryptosporidium sp . em fezes de caes foi de 1% (2/200). Os dois animais eram femeas e SRD, com idade de 60 a 90 dias; uma e de origem residencial e a outra foi resgatada do Centro de Controle de Zoonoses. Embora a ocorrencia tenha sido baixa, infeccoes por Cryptosporidium spp. foram detectadas por metodos moleculares em caninos deste estudo. SUMMARY The protozoan Cryptosporidium spp. is considered an important pathogen in terms of public health (CIELOSZYK et al., 2012), and is mainly associated with low socioeconomic status and poor sanitation conditions (ASSIS et al., 2013). Dogs eliminate fecal oocysts, with or without diarrhea, and are considered as potential sources of human infection (WANG et al., 2012), although this has not been confirmed experimentally (BOWMAN & LUCIO-FORSTER, 2010; UEHLINGER et al., 2013). This study aims at detecting infection by Cryptosporidium spp. in fecal samples from puppies by nested-polymerase chain reaction (Nested-PCR). The samples, previously identified and frozen, were intended for molecular investigation. DNA extraction was performed using the QIAamp DNA Stool kit (Qiagen) and the nested-PCR technique was used for the amplification of gene fragments of the 18S subunit of ribosomal RNA. A total of 200 dogs were examined, 100 males and 100 females, 111 of defined breed and 89 mongrel (non-defined breed, NDB). Of these, 81, 43, 48 and 28 puppies were up to two, two to three, three to six and six to twelve months old respectively. Regarding their origin, the animals were from the municipalities of Aracatuba and Votuporanga, SP, and 126 were household pets; 11 were kept in zoonosis centers; 50 were from Pet Shops; 12 from a breeding farm and one (0.5%) stray pet that had been adopted. The sample prevalence of Cryptosporidium sp. in dog feces was 1% (2/200). Of the two female mongrel animals, aged 60-90 days, one was a pet and the other was rescued from a zoonosis center. Although the occurrence was low, infection by Cryptosporidium spp. was detected in two puppies using molecular techniques.
Databáze: OpenAIRE
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