Caracterização molecular de Cryptosporidium em bezerros de assentamentos rurais da região Noroeste do estado de São Paulo, Brasil

The study was conducted on 25 properties of the settlements Sao Jose I and Salvador, located in the municipalities of Brejo Alegre and Birigui, in the State of Sao Paulo, Brazil. A record of variables was elaborated and included data such as gender, breed and age of the animals. A total of 231 stool samples were collected from bovines aged one to six months, 128 being females and 103 males, 131 crossbred and 100 Holstein. Among the 231 samples, 17 (7.36%) were positive for Crvptosporidium spp. both by malachite green negative staining and by nested-PCR. Of the 17 positive samples, 14 were sequenced in agarose gel. These sequences were detected between 99% and 100% of genetic similarity for the following species. One sequence was similar to C. parvum (AB513880.1), one to C. bovis (MF074602.1), two to C. ryanae (KT922233.1), one to C. felis (KM977642.1) and nine were similar for C. andersoni reference MF350628. C. andersoni was found in animals aged 2-6 months, an age group which is different from those described by several authors. The presence of C. parvum indicates that the calves in the studied region should be considered a potential source for zoonotic transmission. For the first time to our knowledge, C. felis was identified in cattle in America401491496FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP2012/13733-3O estudo foi realizado num total de 25 propriedades localizadas nos assentamentos São José I e Salvador, situados nos municípios de Brejo Alegre e Birigui, no estado de São Paulo. Um registro de variáveis foi elaborado, incluindo dados como sexo, raça e idade dos animais. Foram colhidas 231 amostras de fezes de bovinos de um a seis meses de idade, sendo 128 fêmeas e 103 machos, 131 mestiços e 100 da raça Holandesa. Entre os 231 bovinos examinados, 17 (7,36%) foram positivos para Cryptosporidium spp. tanto pela coloração negativa de verde malaquita como pela nested-PCR. Das 17 amostras positivas, 14 apresentaram amplificação pela eletroforese em gel de agarose suficiente para fazer o sequenciamento de DNA. Essas sequências foram detectadas similaridade genética entre 99% e 100% com as seguintes espécies. Uma sequência foi semelhante com C. parvum (referência: AB513880.1), uma com C. bovis(MF074602.1), duas com C. ryanae (KT922233.1), uma com C. felis (KM977642.1) e nove foram semelhantes com C. andersoni (MF350628). O estudo caracteriza a presença do Cryptosporidiumspp. em bovinos oriundos de propriedades produtoras de leite na região Noroeste do estado de São Paulo, sendo o C. andersoni a espécie mais prevalente nesses animais, principalmente em uma faixa etária diferente das descritas por diversos autores. A presença de C. parvum indica que os bezerros da região estudada devem ser considerados como uma fonte potencial de oocistos de espécies zoonóticas.Identificamos com ineditismo o C. felis em bovinos na América, o que corrobora outros estudos realizados na Polônia e Espanha e evidencia a presença de espécies de Cryptosporidium em fezes em hospedeiros não naturais. Palavras-chave: Bezerros. Caracterização molecular. Criptosporidiose. Nested-PCR.Protozoan of the genus Cryptosporidium are obligate intracellular coccidia with a cosmopolitan distribution. Biological data and various molecular genotyping tools indicate that the genus Cryptosporidium comprises about 30 valid species and more than 70 genotypes that can infect fishes, amphibians, reptiles, birds, and mammals (NAKAMURA; MEIRELES, 2015).Among the named species, 13 have been described in cattle. Of these, nine present zoonotic potential, being more prevalent in humans: Cryptosporidium hominis, Cryptosporidium parvum, Cryptosporidium meleagridis, Cryptosporidium cuniculus and, occasionally,Cryptosporidium muris, Cryptosporidium suis, Cryptosporidium ubiquitum, Cryptosporidium felis(BORNAY-LLINARES et al. 1999; CARDONA et al., 2015) and Cryptosporidium canis (KVÁČ et al., 2016).Cattle are mainly infected with C. parvum, Cryptosporidium bovis, Cryptosporidium ryanaeand Cryptosporidium andersoni (XIAO, 2010). Of these, only the first is of zoonotic importance, being a concern for public health, since human cryptosporidiosis can be acquired from infected calves (LEARMONTH et al., 2004; SOPWITH et al., 2005; DEL COCO et al., 2014). These four species of Cryptosporidium are frequently diagnosed in cattle of different ages. Among these, C. parvumis the most prevalent in young pre-weaning calves (less than two months old) and shows low host specificity, and some genotypes are considered to have high zoonotic potential (DIXON et al., 2011).During the post-weaning phase (three to 11 months of age) a reduction in the prevalence of C. parvum is observed, however, in this stage an increase of C. bovis and C. ryanae infections is noticed. The latter two species are not considered zoonotic and show high specificity for bovine. C. andersoni is the species that mainly infects post-weaned calves and adult cattle. These parasitic species can infect other ruminants and rarely humans (DIXON et al., 2011).Due to the ability of this protozoan to infect different hosts and its constant presence in the environment, humans can acquire the infection in several ways, such as through ingestion of food and water contaminated with oocysts, direct contact with infected people (anthroponotic) or animals (zoonotic). Increases in environmental contamination and infected reservoirs assist in zoonotic transmission, as human cryptosporidiosis is also caused by certain species that also affect ruminants. The contact of humans with the feces of these animals that contains oocysts or contaminated water constitutes routes of transmission to humans (DEL COCO et al., 2014

Conscientização para adoção de boas práticas sanitárias em bem-estar animal no manejo das mastites do rebanho bovino: Miriam Yumi Makatu. -

Aimed at verifying whether the adoption of good agricultural practices is a good tool for the identification of animals with bovine mastitis, and educating the producer of rural settlements about the importance of implementing good agricultural practices related to welfare. The producers were oriented and received educational material and utensils made from recyclable material to the achievement of good health practices. For evaluation of the physiological profile, biological samples such as blood, feces and milk were collected, and for the assessment of sanitation and welfare, a questionnaire was employed. A total of 79 mammary quarters from 20 cows were evaluated. Of the 36 negative quarters for California Mastitis Test, 12 were positive for bacteria, with Staphylococcus being the most frequently isolated. The results indicated that animal welfare was compromised by the incidence of subclinical mastitis, infestation with ectoparasites and the absence of prophylactic measures. It is concluded that the use of good agricultural practices is suitable for the identification of bovine mastitis, however, there is lack of knowledge regarding good practices, mainly related to preventive management and awareness of producers about the importance of implementing good agricultural practices in the dairy activity, which are key to improving animal welfare conditions, productivity and quality of milkCoordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES