Submitted by GUILHERME APARECIDO FIM JÚNIOR null (guilhermejunior10@hotmail.com) on 2016-02-17T01:43:30Z No. of bitstreams: 1 DISSERTAÇÃO__Versão final_PRONTA.pdf: 1645895 bytes, checksum: 10c9b401bd06621d8b3c2c0f30593583 (MD5) / Approved for entry into archive by Ana Paula Grisoto (grisotoana@reitoria.unesp.br) on 2016-02-17T16:39:09Z (GMT) No. of bitstreams: 1 fimjunior_ga_me_jabo.pdf: 1645895 bytes, checksum: 10c9b401bd06621d8b3c2c0f30593583 (MD5) / Made available in DSpace on 2016-02-17T16:39:09Z (GMT). No. of bitstreams: 1 fimjunior_ga_me_jabo.pdf: 1645895 bytes, checksum: 10c9b401bd06621d8b3c2c0f30593583 (MD5) Previous issue date: 2015-12-17 / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Objetivou-se estabelecer a etiologia infecciosa da mastite subclínica ovina em duas diferentes fases de lactação e a frequência desta enfermidade nas diferentes raças. Além disso, buscou-se definir as características diagnósticas da contagem de células somáticas (CCS) para o diagnóstico da mastite subclínica ovina, adotando diferentes classes de contagens de CCS, assim como estabelecer um valor de triagem para a CCS como método diagnóstico. O trabalho experimental foi realizado em um rebanho formado por 407 ovelhas das raças Santa Inês, Texel, Ile de France e Dorper, localizado na Embrapa Pecuária Sudeste em São Carlos, São Paulo. O California Mastitis Testis (CMT) foi realizado para o diagnóstico preliminar da mastite. Após a antissepsia dos tetos, amostras de leite foram colhidas no início de lactação e ao desmame, em duas parições consecutivas para confirmação da etiologia infecciosa da doença por meio de análises microbiológicas e para a determinação da CCS. Para cada uma das raças os valores de CCS foram divididos em quatro classes. O total de 1457 metades mamárias foi investigado, das quais 203 (13,9%) apresentaram mastite subclínica. Estafilococos coagulase-negativos (ECN) foram os agentes etiológicos com maior ocorrência isoladamente e em associação, 58,1% e 60,6%, respectivamente, seguidos por Streptococcus spp. (19,7%). Dentre os ECN, a espécie encontrada com maior frequência foi Staphylococcus xylosus (87,7%). A raça Santa Inês apresentou significativamente maior número de casos de mastite subclínica (p<0,05). Boas e ótimas concordâncias para associação entre CMT e CCS foram obtidos paras todas as raças, exceto Ile de France. Diferentes pontos de corte para CCS foram determinados para as raças estudadas em ambas as fases de lactação. Maiores valores de sensibilidade e especificidade para a CCS foram obtidos para as raças Ile de France e Dorper na lactação, enquanto ao desmame destacaram-se as raças Texel e Dorper. Não houve concordâncias ideais ao comparar as reações de CMT e CCS para a triagem da mastite subclínica. Os resultados do teste variaram de acordo com a raça ovina. A CCS pode ser utilizada para triagem da mastite subclínica em ovelhas de corte de acordo com a prevalência da doença em cada raça, auxiliando técnicos e produtores a melhorar o controle da enfermidade nos rebanhos. / This study aimed to establish the infectious etiology of subclinical mastitis in two different stages of lactation and the frequency of this disease in different breeds sheep. Furthermore, we sought to define the diagnostic characteristics of somatic cell count (SCC) for the diagnosis of subclinical mastitis in these sheep breeds, adopting different classes of SCC, and establish a screening value for SCC as diagnostic method. The experimental work was carried out in a herd composed of 407 sheep of Santa Inês, Texel, Ile de France and Dorper breeds, located in the Embrapa Southeast Livestock, São Carlos, São Paulo. The California Mastitis Test (CMT) was performed for a preliminary diagnosis of mastitis. After antisepsis of the teats, milk samples were collected in early lactation and weaning in two consecutive calvings for confirmation of infectious etiology of the disease by microbiological analysis and the determination of SCC. For each breed, CCS values were divided into four classes. We investigated 1,457 mammary glands, from which 203 (13.9%) had subclinical mastitis. Coagulase-negative staphylococci (CNS) were the etiological agents with higher occurrence isolated or in an association, 58.1% and 60.6%, respectively, followed by Streptococcus spp. (19.7%). Among the CNS, the most frequent species was Staphylococcus xylosus (87.7%).The Santa Inês breed had a significantly higher number of cases of subclinical mastitis (p < 0.05). Good and excellent concordances to the association between CMT and SCC were obtained for all breeds except Ile de France. Different cutoff points to SCC were determined for the studied breeds in both stages of lactation. Higher sensitivity and specificity of SCC cutoff points were obtained for the Ile de France and Dorper breeds during lactation, where as weaning stood out the Texel and Dorper breeds. There were no complete concordances to compare CMT and SCC reactions to the screening of subclinical mastitis. The test results varied according to the breed of sheep. The SCC can be used for screening of subclinical mastitis in sheep according to the prevalence of the disease in each breed, assisting technicians and producers to improve the control of the disease in herds. / FAPESP: 2013/23054-9

Hajj, the annual Muslim pilgrimage to Mecca, Saudi Arabia is a unique mass gathering event that attracts approximately 3 million pilgrims from around the globe. This diverse pilgrim population coupled with the nature of the performed activities raise major public health concerns in the host country with potential global implications. Although gastroenteritis and diarrhea are common among the pilgrims performing Hajj, the microbial etiologies of these infections are still unknown. We used molecular and antigenic approaches to identify the main pathogens associated with Hajj diarrhea. 544 fecal samples from pilgrims suffering from diarrhea whilst performing Hajj during three consecutive seasons (2011-2013) and 99 control samples from 2011 were screened for 16 pathogens that include bacterial, parasitic and viral etiologies that are commonly associated with diarrheal infections. At least one of the screened pathogens could be detected in 42% (n=228) of the samples from the diarrheal cases. Bacteria were the main agents detected in 83% (n=189) of the positive samples, followed by viral and parasitic agents detected in 6% (n=14) and 5% (n=12) respectively. We have also standardized a 16S-based metagenomic approach to identify the gut microbiome in diarrheal cases and non-diarrheal controls in 76 samples. Also, we have standardized a shotgun metagenomics protocol for the direct characterization (diagnosis) of enteric pathogens without cultivation. This approach was used successfully to identify viral (adenovirus) and bacterial causes of Enterotoxigenic E. coli diarrhea from Hajj samples. The findings in this study fill in clear gaps in our knowledge of the etiologies associated with diarrheal infections during Hajj. Foodborne bacteria were the major contributors to Hajj-diarrheal infections. This was coupled with the increased incidences of antimicrobial resistance loci associated with the identified bacteria. These findings would help the public health policy makers to develop and introduce appropriate public health measures to improve the food safety during Hajj.