Um total de 107 S. aureus isolados de casos de mastite, glândulas portadoras, pele do úbere, ordenhadores e insufladores, em rebanhos de São Paulo e Pernambuco, foram tipados por técnicas moleculares PCR-RFLP do gene coa e PCR de spa distinguiram seis perfis. Todas as amostras amplificaram genes coa, spa, icaA e 69% produziram biofilme glicose-induzido in vitro. PFGE identificou 31 perfis e 12 linhagens. Uma linhagem foi predominante (P < 0,0001) e amplamente disseminada em ambas as regiões. Um mesmo perfil foi isolado de mastite clínica, subclínica e portadoras. Houve heterogeneidade genética entre isolados de fazenda. Isolados de origem humana e animal constituíram populações distintas. Poucos isolados de leite, insufladores e pele do úbere tiveram igual perfil. Uma amostra extramamária, 77% dos isolados de leite e. 99% de S. aureus de portadoras produziram biofilme. Não foi detectada correlação entre produção de biofilme e CCS. O isolamento sucessivo do mesmo perfil de PFGE de glândulas assintomáticas por mais de 16 dias caracterizou o estágio de portador. / A total of 107 S. aureus isolated from bovine milk, udder skin, milkers and milking machine, from São Paulo and Pernambuco herds was typed by molecular techniques. PCR-RFLP coa gene and PCR spa gene distinguished six amplicons. All strains amplified coa, spa, icaA genes and, 69% produced in vitro glucose-induced biofilm. PFGE identified 31 pulsotypes, 12 lineages. One of the lineages was predominantly isolated (P<0.0001) and widely disseminated. A same pulsotype was isolated from clinical and, subclinical mastitis as well as from carriers. There was genetic heterogeneity among isolates from the herds. Strains from human and animal origin were genetically different. Few isolates from milk, milking machine and udder skin showed similar pulsotype. An extramammary strain, 77% of the milk isolates and, 99% of the S. aureus isolated from carriers produced biofilm. It was not detected any correlation between SCC and biofilm production. The successive isolation during more than 16 days of a same pulsotype from the asymptomatic glands characterized the carrier status.

Os antimicrobianos são amplamente utilizados no tratamento e prevenção de doenças do gado leiteiro. A ocorrência de resíduos de gentamicina, resultante principalmente do tratamento da mastite bovina é uma das principais causas o aparecimento de resíduos no leite. Neste estudo utilizando como técnica LC/MS-MS, foram analisadas 48 amostras de leite adquiridas em estabelecimentos comerciais, apenas uma apresentou resíduos de gentamicina no valor de 0, 112 ug/mL, superior ao LMR de 100 &#181;g/ml estabelecido pela União Européia (UE), já para o leite in natura, das 55 amostras analisadas, quatro amostras foram positivas apresentando os valores 0,155 ug/mL, 0,192 ug/mL, 0,130 ug/ml e 0,167 ug/mL, todos acima do LMR estabelecido pela União Européia (UE). O método apresentou limite de quantificação de e 0,1 ng/mL, linearidade de 0,1-1500 ng/mL e coeficientes de determinação de 0,992557, recuperação de 91,5 % e com precisão e exatidão adequadas. Conhecer a dimensão da exposição da população a esses compostos é de fundamental importância para promover as ações de controle visando à proteção do consumidor. Desta forma as políticas de inspeção e qualidade do leite criadas pelo governo devem ser aplicadas e servir de garantia para o consumo seguro deste alimento pela população. / The antibiotics are widely used to treat and preventions diseases for dairy cattle. The occurrence of residues of gentamicin, resulting mainly from treatment of bovine mastitis are a major cause the appearance of residues in milk. In this study using the technique LC/MS-MS, were analyzed 48 samples of milk purchased in shops, only one sample present residues of gentamicin showing the value 0,112 ug/ml, exceeding the MRLs of &#181;g/ml established by the European Union (EU) already for milk in nature, from 55 samples, four samples were positive showing the values 0,155 ug/mL, 0,192 ug/mL, 0,130 ug/ml and 0,167 ug/mL, All above the MRLs set by the European Union (EU). The method presented limit of quantification, and 0.1 ng/mL, linearity of 0,1-1500 ng/mL and coefficients of determination of 0.992557, recovery of 91.5% and with precision and accuracy appropriate. know the size of population\'s exposure to these compounds is of fundamental importance to promote measures to control aimed at protecting the consumer. Thus the politics of inspection and quality of milk created by the government should be implemented and serve as a guarantee for the safe consumption of food by the population.

Made available in DSpace on 2014-07-29T15:07:41Z (GMT). No. of bitstreams: 1 Eveline_Xavier.pdf: 537505 bytes, checksum: f46fc6f4fae888045c26180cb3ce26b0 (MD5) Previous issue date: 2008-08-05 / Mastitis is the most frequent disease in dairy cows, it causes great economic losses in herds and dairy industries and represents a hazard for public health. Among the etiologic agents identified, there is a predominance of bacteria species. However, several other groups of microorganisms may be present, one of those is a unicellular alga of the Prototheca genus, the Prototheca zopfii, an environmental agent of bovine mastitis. The infected cows usually have granulomatous changes in mammary tissue. Moreover microorganisms do not respond to routine therapy, resulting in the elimination of the infected animals. The main purposes of this study was to determine the occurrence of Prototheca zopfii on bulk tank milk in the state of Goiás and identify the limit of detection for Prototheca zopfii by two different approaches. We also wanted to know the samples pre-incubated with small concentration of Protoheca zopfii permits us to obtain positive growth in microbiological culture. We analyzed 473 samples from farms with high somatic cell count patterns but we did not detect Protoheca zopfii in any of those samples. The limit of sensitivity of the Prototheca zopfii was analyzed, in the approach of isolation the limit of detection was between 130 to 3,400 UFC/mL. In the approach of tally the limit was between 25 to 300 UFC/mL. It wasn t observed significant difference (p>0,05) between the approaches analyzed. However, significant statistical difference (p<0,05) occurred between the five types analyzed. After the pre-incubation at 37ºC/24h, the limit of sensibility was analyzed by the approach isolation, the limit was 30 UFC/mL, what means that the pre-incubation is so important to decrease the possibility of occurrence of false negative / Entre as enfermidades de grande preocupação para os bovinos leiteiros encontram-se as mastites, incluindo aquelas causadas por Prototheca spp., um agente ambiental, relacionado à mastite bovina que destaca-se pela gravidade das lesões que causam no tecido mamário e pela resistência a ações terapêuticas. Objetivou-se com o presente estudo determinar a ocorrência de Prototheca zopfii no leite cru refrigerado no estado de Goiás, identificar o limite de detecção deste patógeno pelos métodos de semeadura para isolamento, e semeadura para contagem e também verificar se a incubação de amostras de leite com baixo número de células da Prototheca zopfii permite a obtenção de resultados positivos na cultura microbiológica. Foram analisadas 473 amostras originárias de propriedades com histórico de elevada contagem de células somáticas e procedeu-se a avaliação do limite de sensibilidade para o agente por meio do método de semeadura para isolamento, cujo limite de detecção correspondeu à faixa de 130 a 3.400 UFC/mL. Quanto ao método de semeadura para contagem o limite estabelecido foi 25 a 300 UFC/mL. Portanto houve diferença estatística (p<0,05) entre os métodos analisados, porém, não houve diferença estatística (p>0,05) entre as cinco cepas analisadas. Após a incubação das amostras por 24 horas a 37ºC verificou-se que o limite de sensibilidade pelo método semeadura para isolamento diminui para 30 UFC/mL, diminuindo-se assim a possibilidade de ocorrência de falso negativo. Em nenhuma das amostras analisadas foi detectada a presença da Prototheca zopfi

Submitted by Cláudia Bueno (claudiamoura18@gmail.com) on 2016-01-28T16:09:59Z No. of bitstreams: 2 Tese - Marília Cristina Sola - 2015.pdf: 1799194 bytes, checksum: 1cc96734092d165f83412bce38881a2c (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2016-01-29T11:32:22Z (GMT) No. of bitstreams: 2 Tese - Marília Cristina Sola - 2015.pdf: 1799194 bytes, checksum: 1cc96734092d165f83412bce38881a2c (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Made available in DSpace on 2016-01-29T11:32:23Z (GMT). No. of bitstreams: 2 Tese - Marília Cristina Sola - 2015.pdf: 1799194 bytes, checksum: 1cc96734092d165f83412bce38881a2c (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Previous issue date: 2015-10-02 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / This study aimed to analyze the health aspects of the mammary gland, whey protein profile, and quality of milk produced by the breeds Curraleiro Pe-Duro and Pantaneiro. We evaluated 226 milk samples from a Curraleiro Pé-Duro herd, in the municipality of Cocalzinho, Goiás, and 107 milk samples from a Pantaneiro herd, belonging to the Nhumirim farm - Embrapa Pantanal experimental unit, municipality of Corumbá, Mato Grosso do Sul, from January 2013 to May 2014. We determined values related to the milk chemical composition (fat, protein, lactose, degreased, and total dry extract) and physical aspects (acidity, pH, electroconductivity). According to the results of this study, we determined the values related to the chemical composition (fat, protein, lactose, total solids, and degreased) and physical aspects of milk (acidity, pH, electroconductivity). Cases of clinical and subclinical mastitis were identified in herds with the involvement of bacteria of the genera Staphylococcus, Streptococcus, Bacillus, Corynebacterium, Escherichia, Serratia, Enterobacter, Klebsiella, Citrobacter, Proteus, Yersinia, and Shigella. There was no presence of antibiotic residues in the milk of both breeds. It was possible to characterize the main proteins present in whey as the concentration, molecular weight, and gel migration time due to microfluidic electrophoresis assays, besides the evidence of mastitis effect on whey proteins regarding the increase in the concentration of albumin, immunoglobulin G, and lactoferrin. The results of this study suggest that Curraleiro Pé-Duro and Pantaneiro showed good conditions of milk quality and sanity of the mammary gland in most animals evaluated, justifying the importance in the conservation of these genetic resources for future contribution to the development of the national livestock. / O presente estudo teve por objetivo analisar os aspectos sanitários da glândula mamária, perfil de proteínas no soro lácteo e a qualidade do leite produzido pelas raças Curraleiro Pé-Duro e Pantaneiro. Foram avaliadas 226 amostras de leite obtidas de um rebanho da raça Curraleiro Pé-Duro, localizados no município de Cocalzinho, Goiás e 107 amostras de leite de bovinos Pantaneiro, pertencentes à Fazenda Nhumirim -unidade experimental da Embrapa Pantanal, município de Corumbá, Mato Grosso do Sul, no período de janeiro de 2013 a maio de 2014. De acordo com os resultados encontrados neste estudo, foi possível determinar os valores relacionados à composição centesimal do leite (gordura, proteína, lactose, extrato seco total e desengordurado) e aspectos físicos (acidez, pH e eletrocondutividade). Foram identificados casos de mastite clínica e subclínica nos rebanhos com o envolvimento dos gêneros bacterianos Staphylococcus, Streptococcus, Bacillus, Corynebacterium, Escherichia, Serratia, Enterobacter, Klebsiella, Citrobacter, Proteus, Yersinia e Shigella. Não foi observada a presença de resíduos de antimicrobianos no leite de ambas as raças. A partir da realização da técnica de eletroforese microfluídica foi possível caracterizar as principais proteínas presentes no soro do leite quanto a concentração, peso molecular e tempo de migração em gel, além de evidenciar a efeito da mastite sobre as proteínas presentes no soro lácteo, a partir da elevação na concentração de albumina, imunoglobulina G e lactoferrina. Os resultados obtidos neste estudo sugerem que as raças Curraleiro Pé-Duro e Pantaneiro apresentaram boas condições quanto à qualidade do leite e sanidade da glândula mamária na maioria dos animais avaliados, justificando a importância na conservação destes recursos genéticos para futura contribuição ao desenvolvimento da pecuária nacional.

Submitted by Luciana Ferreira (lucgeral@gmail.com) on 2014-12-22T14:31:16Z No. of bitstreams: 1 Dissertação - Maria Juíva Marques de Faria - 2014.pdf: 3781718 bytes, checksum: 5a98e4c3d48dde6e80b58427a1a73012 (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2015-01-26T13:13:18Z (GMT) No. of bitstreams: 1 Dissertação - Maria Juíva Marques de Faria - 2014.pdf: 3781718 bytes, checksum: 5a98e4c3d48dde6e80b58427a1a73012 (MD5) / Made available in DSpace on 2015-01-26T13:13:18Z (GMT). No. of bitstreams: 1 Dissertação - Maria Juíva Marques de Faria - 2014.pdf: 3781718 bytes, checksum: 5a98e4c3d48dde6e80b58427a1a73012 (MD5) Previous issue date: 2014-05-07 / Bovine mastitis is an inflammatory/infectious process of the mammary gland, its etiology is complex and multivariate. The most common mastitis has bacterial origin. Because of the high cost to treat this disease, and the bacterial resistance to antibiotics, beyond the losses due to the milk discard or the presence of drug residues in milk, studies are necessary to find out alternative methods to circumvent these problematic. The medicinal plants with antimicrobial activity have been making room in veterinary medicine with the objective to reduce the impact and the use of drugs indiscriminately, as well as maintain the integrity of the animal and control the dissemination of resistant bacterial lineage. Therefore, the aim of this work was characterize the oleoresin Copaifera spp., verify the antimicrobial activity in vitro of oleoresin and essential oil of Copaifera spp. in combating against aerobic and facultative isolated bacteria from cows’ milk with subclinical grade III mastitis diagnostic and evaluate preliminarily, in vivo, phytotherapic formulation composed of Copaifera spp. The tests for characterization of the oleoresin, confirmed its authenticity and quality, considered appropriate for use. The analyzes performed by gas chromatography coupled to mass spectrometry (GC/MS) and electrospray ionization Fourier transformed íonic cyclotron resonance mass spectrometry (ESI FT-ICR MS) identified sesquiterpenes and diterpenes present in the essential oil and oleoresin of Copaifera spp., respectively. The technique of High Performance Liquid Chromatography (HPLC) for sesquiterpenes, especially for β-caryophyllene, can be used in quality control of essential oil and oleoresin of Copaifera spp., as well as in phytotherapic formulation using oleoresin of Copaifera spp. The oleoresin showed better antimicrobial activity than the essential oil of Copaifera spp. (ρ<0,0001) in combating the 55 isolated bacteria. The antibiogram test of the three phytotherapic formulations demonstrated that the oleoresin had antimicrobial activity against the isolated microorganisms. The preliminary evaluation, in vivo, of a phytotherapic formulation containing 20% oleoresin of copaiba was neither effective nor safe, because it intensified the inflammatory process, requiring further tests, such as cytotoxicity and irritability of the raw material plant and of the vehicle used. Key words: Bovine Mastitis. Oleoresin. Essential Oil. Copaifera spp. Minimum Inhibitory Concentration. ESI FT-ICR MS. / Mastite bovina é um processo inflamatório/infeccioso da glândula mamária, de etiologia complexa e multivariada, sendo a de origem bacteriana a mais frequente. Devido aos elevados custos dos tratamentos dessa enfermidade, resistência bacteriana aos antibióticos, além dos prejuízos inerentes ao descarte do leite ou à presença de resíduos de medicamentos no mesmo, faz-se necessário estudos que busquem métodos alternativos para contornar essa problemática. As plantas medicinais com potencial antimicrobiano vêm ganhando espaço na medicina veterinária com o objetivo de reduzir o impacto e o uso de medicamentos de forma indiscriminada, bem como manter a integridade do animal e controlar a disseminação de linhagens bacterianas resistentes. Diante disso, o objetivo desse trabalho foi caracterizar a oleorresina de Copaifera spp., verificar atividade antimicrobiana in vitro da oleorresina e do óleo essencial de Copaifera spp. frente às bactérias aeróbias e facultativas isoladas de leite de vacas diagnosticadas com mastite subclínica grau III e avaliar preliminarmente, in vivo, formulação fitoterápica à base de Copaifera spp. Os testes de caracterização da oleorresina de Copaifera spp. demonstraram sua autenticidade e qualidade, a qual foi considerada adequada para o uso. As análises realizadas por cromatografia gasosa acoplada à espectrometria de massas (CG/EM) e por espectrometria de massas com ionização por electrospray de ressonância ciclotrônica de íons por transformada de Fourier (ESI FT-ICR MS) identificaram os sesquiterpenos e diterpenos presentes no óleo essencial e na oleorresina de Copaifera spp., respectivamente. A técnica Cromatografia Líquida de Alta Eficiência (CLAE) para sesquiterpenos, em especial para β-cariofileno, pode ser utilizada no controle de qualidade de oleorresina e óleo essencial de Copaifera spp., bem como em formulação fitoterápica utilizando oleorresina de Copaifera spp. como matéria-prima vegetal. A oleorresina apresentou melhor atividade antimicrobiana do que o óleo essencial de Copaifera spp. (ρ<0,0001) frente às 55 bactérias isoladas. O teste de antibiograma das três formulações fitoterápicas demonstrou que a oleorresina de copaíba reproduziu a sua atividade antimicrobiana frente aos micro-organismos isolados. A avaliação preliminar, in vivo, de uma formulação fitoterápica contendo 20% oleorresina de copaíba não foi eficaz e nem segura, pois intensificou o processo inflamatório, havendo necessidade de testes futuros, como de citotoxicidade e de irritabilidade dessa matéria-prima vegetal e do veículo utilizado.

The present study aimed to evaluate bacterial isolates and the microbiome of dairies. The specific aims were: to characterize Staphylococcus spp. isolated from mastitic milk, to evaluate the presence of Lactococcus in mastitic milk as a potential causative agent of mastitis, to evaluate the association between microbiome and milk quality parameters, and to characterize Staphylococcus spp. isolated from production lines of Minas Frescal cheese. The detection of genes encoding virulence factors (enterotoxins (sea, seb, sec, sed, see, seg, seh, sei, selj, selk, sell, selm, seln, selo, selp, seIq, ser, ses, set, selu, selv, and selx), hemolysins (hla, hlb, hld, hlg, and hlgv), exfoliative toxins (eta, etb, and etd), Panton-Valentine leukocidin (pvl), and toxic shock syndrome toxin (tst)), genes encoding antibiotic resistance (resistance to tetracycline (tetK, tetL, and tetM), erythromycin (ermA, ermB, and ermC), methicillin (mecA and mecC), and tobramycin (ant(4\')-Ia)), molecular typing (spa, SCCmec, and agr types), and phenotyping regarding antibiotic resistance were performed in staphylococci isolates from mastitic milk, and from cheese processing plant samples. Staphylococcus aureus was identified in the majority of isolates from both origins. Several virulence factor genes were detected. The distribution of genes encoding staphylococcal enterotoxins (85.0% - 85.7% of isolates were positive for one or more enterotoxin gene) was highlighted and the gene related to H toxin was the most prevalent. Methicillin-resistant Staphylococcus aureus were identified in isolates from mastitic milk (4.1%) and cheese processing (6.0%); the genotyping and phenotyping of these isolates were described. t605 had the highest frequency in the S. aureus population studied. In mastitic milk, Lactococcus was suggested as the causative agent of an outbreak of mastitis in a dairy farm. Using next generation sequencing, the abundance of Lactococcus was observed in microbiome samples. Bacterial isolation and DNA sequencing confirmed the presence of Lactococcus lactis and Lactococcus garvieae. The microbiome of environmental samples and bulk tank milk from the dairy farm showed the Lactococcus genus among the most common bacterial taxa, suggesting other sources of this genus. Regarding milk quality parameters, the microbiome of bulk tank milk from several dairy farms was associated with somatic cell count and bacterial count. The core microbiome was described and many genera of importance were identified. Among the associations performed between microbiome and milk quality parameters, the identification of Streptococcus in samples classified with high somatic cell count and high bacterial count was highlighted. Several bacterial taxa with relative abundance significantly higher in samples classified as high and low cell count and bacterial count were shown. Real-time polymerase chain reaction was also performed associated with bacterial diversity, bacterial taxa, and bacterial count. These findings highlight the need to control and prevent bacterial contamination in the dairy industry, from herd to consumers. / O presente estudo apresentou como objetivo avaliar isolados bacterianos e microbioma de lácteos. Os objetivos específicos foram: caracterizar Staphylococcus spp. isolados de leite de vacas com mastite, avaliar a presença de Lactococcus em leite de vacas com mastite como um potencial agente causador de mastite, avaliar a associação entre microbioma de leite de tanque e parâmetros da qualidade de leite, e caracterizar Staphylococcus spp. isolados de linhas de processamento de queijo Minas frescal. A detecção de genes codificadores de fatores de virulência (enterotoxinas (sea, seb, sec, sed, see, seg, seh, sei, selj, selk, sell, selm, seln, selo, selp, seIq, ser, ses, set, selu, selv, e selx), hemolisinas (hla, hlb, hld, hlg, e hlgv), toxinas exfoliativas (eta, etb e etd), leucocidina de Panton-Valentine (pvl), toxina da síndrome do choque tóxico (tst)), genes codificadores de resistência a antibióticos (resistência a tetraciclina (tetK, tetL e tetM), eritromicina (ermA, ermB e ermC), meticilina (mecA e mecC) e tobramicina (ant(4\')-Ia)), tipagem molecular (spa, SCCmec e agr types), e fenotipagem quanto à resistência a antibióticos foram realizadas em estafilococos isolados de leite de vacas com mastite e de amostras de planta de processamento de queijo. Staphylococcus aureus foi identificado na maioria dos isolados de ambas as origens. Diversos genes de fatores de virulência foram detectados, com destaque para a distribuição de genes codificadores de enterotoxinas estafilocócicas (85,0%-85,7% dos isolados foram positivos para um ou mais genes codificadores de enterotoxinas), sendo o gene relacionado com a toxina H o mais frequente. Staphylococcus aureus meticilina resistente foram identificados em isolados de leite de vacas com mastite (4.1%) e em processamento de queijo (6.0%); o perfil genotípico e fenotípico destes isolados foram descritos. t605 foi o mais freqüente na população de S. aureus estudada. Em leite de vacas com mastite, Lactococcus foi sugerido como o agente causador de um surto de mastite numa fazenda leiteira. Usando sequenciamento de nova geração, a abundância de Lactococcus foi observada no microbioma das amostras. O isolamento e sequenciamento de DNA confirmaram a presença de Lactococcus lactis e Lactococcus garvieae. O microbioma de amostras ambientais e de leite de tanque da fazenda mostrou o gênero Lactococcus entre os mais comuns, sugerindo outras fontes deste gênero. Contemplando parâmetros da qualidade de leite, o microbioma de leite de tanque de várias fazendas leiteiras foi relacionado com contagem de células somáticas e contagem bacteriana. O core microbiome foi descrito e muitos gêneros bacterianos de importância foram identificados. Dentre as análises realizadas associando microbioma com parâmetros da qualidade de leite, foi destacada a identificação de Streptococcus em amostras classificadas com alta contagem de células somáticas e alta contagem bacteriana. Diversos táxons bacterianos com abundância relativa significativamente maior em amostras classificadas com alta e baixa contagem de células somáticas e contagem bacteriana foram mostrados. Reação em cadeia da polimerase em tempo real também foi realizada e associada com diversidade bacteriana, táxons bacterianos e contagem bacteriana. Estes levantamentos confirmam a necessidade de controlar e prevenir a contaminação bacteriana na indústria de lácteos, do rebanho leiteiro até os consumidores.

Made available in DSpace on 2014-07-29T15:13:40Z (GMT). No. of bitstreams: 1 dissertacaoMarcoAntonio.PDF: 791526 bytes, checksum: 7a8a829a082433a7f23c3880ef18467d (MD5) Previous issue date: 2008-12-19 / The objective of the research were evaluate cooled raw milk gotten in dairy properties of the Southwest Goiano in the periods rainy and dry of 2008. The samples were collected of individual producers where the storage in bulk tanks were carried for until 72 hours, with 24-hour intervals. Somatic cells count, total bacterial count and centesimal composition were carried in the Laboratório de Qualidade do Leite of the Centro de Pesquisa em Alimentos of the Escola de Veterinária of the Universidade Federal de Goiás. The microbiological analysis and titratable acidity were carried in the Laboratories of the Unidade de Agroindústria of the Instituto Federal de Educação, Ciência e Tecnologia Goiano Campus Rio Verde GO. Data were submitted to the variance analysis and the analyzed factors were: period, type of milking and storage time, in entirely casualized delineation and factorial arrangement 2 x 2 x 4. The comparison of period and type of milking was carried through by means of test F of the variance analysis. The storage time was analyzed by means of regression models. Software SISVAR was used for analysis. The results of the physical-chemical composition were in accordance with the legislation. The total bacterial count of cooled raw milk to the 24 hours of storage was above of the limit of the legislation. The count of psychrotrophic, psychrotrophic proteolytic and Pseudomonas spp., was bigger in the rainy period. / O objetivo da pesquisa foi avaliar o leite cru refrigerado obtido em propriedades rurais do Cerrado Goiano nos períodos chuvoso e seco de 2008. As amostras foram coletadas de produtores individuais onde o armazenamento em tanques de expansão era realizado por até 72 horas, com intervalos de 24 horas. As análises eletrônicas (contagem de células somáticas, contagem bacteriana total e composição centesimal) foram realizadas no Laboratório de Qualidade do Leite do Centro de Pesquisa em Alimentos da Escola de Veterinária da Universidade Federal de Goiás. As análises microbiológicas e de acidez titulável foram realizadas nos Laboratórios de Microbiologia e Processamento de Leite e Derivados da Unidade de Agroindústria do Instituto Federal de Educação, Ciência e Tecnologia Goiano Campus Rio Verde GO. Os dados foram submetidos à análise de variância, e os fatores analisados foram: período (chuvoso ou seco), tipo de ordenha (manual ou mecânica) e tempo de estocagem (zero, 24, 48 e 72 horas), em delineamento inteiramente casualizado e arranjo fatorial 2 x 2 x 4. A comparação do período e tipo de ordenha foi realizada por meio do teste F da análise de variância. Os tempos de estocagem foram analisados por meio de modelos de regressão. Para atender as pressuposições da análise de variância a contagem de psicrotróficos, psicrotróficos proteolíticos, Pseudomonas spp., CCS e CBT foram transformadas por meio do logaritmo na base 10. Foi utilizado o Software SISVAR para análise. A contagem bacteriana total do leite cru refrigerado às 24 horas de armazenamento ficou acima do limite exigido pela legislação. A contagem de psicrotróficos, psicrotróficos proteolíticos e Pseudomonas spp., foi maior no período chuvoso. Deverão ser alertados, produtores e indústrias para a obtenção e processamento de leite com qualidade higiênico-sanitária adequada.

Introduction : Le cancer de l’ovaire est le cancer gynécologique le plus fatal dans le monde et est associé à un fardeau économique considérable pour les systèmes de santé publique, les patients et leurs familles. Actuellement, la prévention de ce cancer passe par l’identification des facteurs de risque, dont l’inflammation. Le double rôle de l’inflammation dans la carcinogenèse (transformation néoplasique et stimulation de la croissance pour l’inflammation chronique, mais également l’inhibition de la croissance pour inflammation aiguë) a déjà été observé au 19ième siècle, par Rudolf Virchow et par l’allemand Bruns, respectivement. Plusieurs preuves suggèrent aussi que le cancer de l’ovaire pourrait être lié à l’inflammation chronique de l’épithélium ovarien d’où l’hypothèse selon laquelle les analgésiques ayant une action anti-inflammatoire comme les anti-inflammatoires non stéroïdiens (AINS) et l’acétaminophène pourraient prévenir le cancer de l’ovaire. Contrairement à l’inflammation chronique, un autre facteur intéressant qui pourrait jouer un rôle sur le cancer de l’ovaire par le biais d’une inflammation aiguë est la mastite puerpérale qui est la forme la plus courante de mastite. Cependant, la littérature existante, examinant l’usage des analgésiques (aspirine, AINS non aspirine et acétaminophène) et le risque de cancer ovarien, est incohérente avec des différences populationnelles (cohortes de naissance différentes) et méthodologiques : variations des définitions de l’utilisation régulière, des variables d’ajustement, mais aussi dans la prise en compte d’une possible causalité inverse. De plus, aucune étude n’a tenté d’évaluer l’association dépendante du temps entre l’utilisation régulière de ces médicaments et le risque de cancer ovarien. Pour la mastite puerpérale pendant l’allaitement, deux articles avaient évalué son association avec le risque de cancer épithélial de l’ovaire (CEO), mais avec des limites méthodologiques : violation de la positivité avec l’inclusion des femmes qui n’ont jamais eu de grossesse et sur-ajustement avec la durée d’allaitement qui est dans le chemin causal. Objectif : Cette thèse visait à atteindre deux objectifs généraux qui sont de fournir de nouvelles preuves concernant les associations entre : 1) l’utilisation régulière d’analgésiques et le risque de CEO ; 2) la mastite puerpérale et le risque de CEO. Méthode : Nous avons utilisé les données d’une étude cas-témoin populationnelle visant à documenter les facteurs pour la prévention du cancer de l’ovaire au Québec (Étude PROVAQ). Cette étude a été menée dans la grande région de Montréal, Canada, de mars 2011 à septembre 2016 avec 498 cas et 908 témoins. Notre approche méthodologique a été effectuée en trois étapes. Premièrement, nous avons utilisé l’ensemble des données de PROVAQ pour l’évaluation des associations entre l’utilisation régulière de types de médicaments analgésiques, et aussi selon l’indication et le risque de CEO. Deuxièmement, à partir des données de PROVAQ, nous avons évalué l’association dépendante du temps entre l’utilisation régulière d’un type de médicaments et le risque de CEO à l'aide d'un indice cumulatif pondéré flexible d'exposition dans des modèles de régression logistique conditionnelle. Enfin, nous avons évalué l’association entre la mastite puerpérale et le risque de CEO chez les femmes allaitantes (174 cas et 431 témoins). La régression logistique a été utilisée pour estimer ces associations. Résultats : Nos résultats suggèrent que l'utilisation régulière d’aspirine et d'AINS non aspirine était inversement associée au CEO avec des rapports de cotes (RC) ajustés de 0,81 (IC à 95 % : 0,57–1,12) et 0,74 (IC à 95 % : 0,54–1,00), respectivement. Pour l'utilisation régulière d'AINS non aspirine, les RCs ajustés des COX-2 non sélective et sélective étaient de 0,73 (IC à 95 % : 0,50–1,00) et de 0,83 (IC à 95 % : 0,48–1,40), respectivement. Des associations similaires ont été observées selon le niveau de durée cumulative à vie ou de quantité cumulative à vie de prises d’aspirine et d’AINS non aspirine. Cependant, les associations entre les types de médicaments analgésiques et le CEO peuvent différer selon leurs indications. Aucune association n’a été trouvée entre le moment de l'utilisation régulière d’un type de médicaments analgésiques au cours des 40 années précédant la date index et le CEO. Aucune association significative n’a été aussi trouvée entre la mastite puerpérale pendant l'allaitement et le CEO (RC = 1,15 ; IC à 95 % : 0,71–1,84). Conclusions : Cette thèse fournit des preuves qui appuient l'hypothèse selon laquelle l'utilisation régulière d'aspirine et d'AINS non aspirine sont inversement associées au CEO. Nos résultats suggèrent également l'importance de considérer les indications d'utilisation lors de l'examen des relations entre les types de médicaments analgésiques et le CEO. Elle n’a pas trouvé d'association entre le moment de l'utilisation régulière d’analgésiques et le CEO mais aussi entre la mastite puerpérale pendant l’allaitement et le CEO. Cependant, notre étude a manqué de puissance. / Introduction: Ovarian cancer is the most fatal gynecological cancer in the world and is associated with a considerable economic burden for public health systems, patients and their families. Currently, the prevention of this cancer requires the identification of risk factors including inflammation. The dual role of inflammation in carcinogenesis (neoplastic transformation and stimulation of cancer growth for chronic inflammation, but also inhibition of cancer growth for acute inflammation) has already been observed in the 19th century, by Rudolf Virchow and by the German Bruns, respectively. Several pieces of evidence also suggest that ovarian cancer could be linked to chronic inflammation of the ovarian epithelium, hence the hypothesis that analgesics with an anti-inflammatory action such as nonsteroidal anti-inflammatory drugs (NSAIDs) and acetaminophen could prevent ovarian cancer. Unlike chronic inflammation, another interesting factor that could play a role in ovarian cancer through acute inflammation is puerperal mastitis which is the most common form of mastitis. However, the existing literature examining the use of analgesics (aspirin, non-aspirin NSAIDs and acetaminophen) and the risk of ovarian cancer is inconsistent with population (different birth cohorts) and methodological differences: variations in definitions of regular use, adjustment variables but also in taking into account a possible reverse causality. In addition, no studies have attempted to assess the time-dependent association between regular use of these drugs and the risk of ovarian cancer. For puerperal mastitis during breastfeeding, two articles had assessed its association with the risk of epithelial ovarian cancer (EOC) but with methodological limitations: violation of positivity with the inclusion of women who never had of pregnancy and over-adjustment with the duration of breastfeeding which is in the causal path. Objective: This thesis aimed to achieve two general objectives which are to provide new evidence regarding the associations between: 1) the regular use of analgesics and the risk of EOC; 2) puerperal mastitis and the risk of EOC. Method: We used data from a population-based case-control study aimed at documenting factors for the prevention of ovarian cancer in Quebec (PROVAQ study). This study was conducted in the greater Montreal area, Canada, from March 2011 to September 2016 with 498 cases and 908 controls. Our methodological approach was carried out in three stages. First, we used the PROVAQ dataset to assess associations between regular use of analgesic drugs types, and also by indication and EOC risk. Second, from PROVAQ data, we evaluated the time-dependent association between regular use of a type of medication and the risk of EOC using a flexible weighted cumulative index of exposure in conditional logistic regression models. Finally, we evaluated the association between puerperal mastitis and the risk of EOC in lactating women (174 cases and 431 controls). Unconditional logistic regression was used to estimate associations between regular use of analgesic drugs types, puerperal mastitis during breastfeeding and EOC risk. Results: Our results suggest that regular use of aspirin and non-aspirin NSAIDs were inversely associated with EOC with adjusted ORs of 0.81 (95% CI: 0.57–1.12) and 0.74 (95% CI: 0.54–1.00), respectively. For regular non-aspirin NSAID use, the adjusted ORs for non-selective and selective COX-2 were 0.73 (95% CI: 0.50–1.00) and 0.83 (95% CI: 0.48–1.40), respectively. Similar associations were observed according to the level of lifetime cumulative duration or lifetime cumulative quantity of aspirin and non-aspirin NSAID. However, the associations between analgesic drug types and EOC may differ according to their indications. No association was found between the time of regular use of any type of analgesic medication in the 40 years prior to the index date and EOC. No significant association was also found between puerperal mastitis during breastfeeding and EOC (OR = 1.15; 95% CI: 0.71–1.84). Conclusions: This thesis provides evidence that supports the hypothesis that regular use of aspirin and non-aspirin NSAIDs are inversely associated with EOC. Our results also suggest the importance of considering indications for use when examining relationships between analgesic drug types and EOC. We found no association between the timing of regular analgesic use and EOC but also between puerperal mastitis during breastfeeding and EOC. However, our study was underpowered.