Einleitung: Euterinfektionen sowohl mit kuh-, vor allem mit umweltassoziierten Streptokokken nehmen bei Milchkuhherden eine dominierende Rolle ein. Ziel der Untersuchungen: Die Beziehungen zwischen der Innerherdenprävalenz (IHP) von subklinischen Mastitiden mit Streptokokken zur Haltungshygiene und zur Melkhygiene wurden erfasst. Dabei standen laktierende Kühe mit Nachweis von äskulinpositiven Streptokokken und Streptococcus agalactiae in Viertelanfangs-gemelksproben im Mittelpunkt. Tiere, Material und Methoden: Von September 2009 bis Dezember 2010 fanden in 34 Thüringer Milchviehherden zweimalig im Abstand von etwa 100 Tagen bakteriologische Untersuchungen der Viertelanfangsgemelke von allen zu diesem Zeitpunkt laktierenden und klinisch eutergesund erscheinenden Kühen statt. Der Medianwert der Herdengröße der laktierenden Kühe betrug 246. Die bakteriologischen Untersuchungen von 81.567 Viertelanfangsgemelken erfolgten entsprechend den Leitlinien der Deutschen Veterinärmedizinischen Gesellschaft durch das Labor des Tiergesundheitsdienstes der Thüringer Tierseuchenkasse. Während des ersten Bestandsbesuchs wurden die Haltungsbedingungen sowie die Melktechnik und die Melkhygiene im Betrieb mittels eines dreiteiligen, strukturierten Fragebogens erfasst. Die Bewertung der Merkmale erfolgte durch die Einteilung in zwei oder drei vorgegebene Kategorien. Mittels multivariabler Varianzanalyse wurde anschließend geprüft, ob eine Beziehung zwischen der IHP intramammärer Infektionen mit Streptococcus agalactiae bzw. äskulinpositiven Streptokokken und dem jeweiligen Risikofaktor besteht. Ergebnisse: Infektionen mit äskulinpositiven Streptokokken traten in allen Herden auf, während Galtstreptokokken in nur sieben der 34 Herden nachgewiesen werden konnten. Der Median der IHP der Infektionen mit äskulinpositiven Streptokokken lag zur Untersuchung 1 bei 8,5 %, zur Untersuchung 2 bei 4,6 %, für Infektionen mit Streptococcus agalactiae bei 4,9 % bzw. 3,1 %. Signifikante Beziehungen zur IHP von subklinischen Mastitiden mit äskulinpositiven Streptokokken oder Streptococcus agalactiae finden sich für die Sauberkeit der Liegeflächen im Bereich der Trockensteher und der kalbenden Kühe, für die Art der Laufflächen im Bereich der Trockensteher und der milchliefernden Kühe und für die regelmäßige Nutzung der Handwaschgelegenheiten durch das Melkpersonal. Für Galt-Nachweise zeigten sich signifikante Beziehungen und niedrige IHP im Zusammenhang mit der Verwendung einer Rohrmelkanlage für die Frischmelker, guter Sauberkeit der Euter der zum Melken kommenden Kühe und der Verwendung einer Tauchdesinfektion als Melkzeugzwischendesinfektion. Für subklinische Mastitiden mit äskulinpositiven Streptokokken gilt dies für Herden, in denen die Kühe der Leistungsherde mit kaum verschmutzten Eutern zum Melken kommen und stark verschmutzte Euter in der Leistungsherde vor dem Melken intensiv gereinigt werden. Schlussfolgerungen: Die Häufigkeit der Infektionen mit äskulinpositiven Streptokokken steht mit der Sauberkeit der Euter in Beziehung. Das Bereitstellen von sauberen und trockenen Liegeflächen und Laufgängen für alle Kühe in der Herde, auch für die trockenstehenden Tiere, trägt zu einer Senkung der IHP intramammärer Infektionen mit umweltassoziierten Streptokokken bei. Dabei geht es um die Vermeidung von Verschmutzungen der Euter. Weiterhin stellen das Melken der frischmelkenden Kühe in einem separaten Melkstand und die Melkzeugzwischendesinfektion durch Tauchen geeignete Maßnahmen zur Unterbindung der Erregerverbreitung von Euterinfektionen mit Streptococcus agalactiae dar.

Ziel der Arbeit war es, die Eignung des Coulter Counters für das Eutergesundheitsmonitoring zu untersuchen. Hierfür wurde, als Voraussetzung für den Einsatz des Coulter Counters in der prozessnahen Milchuntersuchung, eine Probenaufbereitungsmethode entwickelt. Diese ermöglicht es durch Überkopfzentrifugation mit hoher Geschwindigkeit (1600 × g; 4 °C; 15 Minuten) die Proben binnen weniger Minuten so aufzubereiten, dass sie im Coulter Counter gemessen werden können. Die Überkopfzentrifugation garantiert dabei die Messbarkeit der Zellen sowohl des Zellpellets auch des Überstands ohne Fettrückstände in der Probe. Es wurde festgestellt, dass sich die Partikelgrößenverteilung im Falle einer Mastitis deutlich verändert. Basierend auf den Partikelgrößenverteilungsdaten konnten mittels eines Trainingsdatensatzes Algorithmen zur Differenzierung des Gesundheitsstatus entwickelt werden. Die Zuordnung des Gesundheitsstatus konnte im Testdatensatz mit einer Spezifität von 100 % für den Status „gesund“, 96,9 % für den Status „subklinisch erkrankt“ und 91,2 % für den Status „erkrankt“ nachgewiesen werden. / The aim of this study was, to examine the suitability of the Coulter Counter for udder health monitoring. Therefore a sample preparation method was developed as precondition for using the Coulter Counter for online Monitoring. The preparation method allows to prepare the samples within a few minutes to make them measurable in the Coulter Counter. For this a high speed centrifugation in overhead position is used (1600 × g; 4 °C; 15 min). This guarantees to measure the cells of the cell pellet as well as of the supernatant, without any fat particles in the sample. It could be shown that the particle size distribution changes during a mastitis. Based on the data of the particle size distribution algorithms to differentiate the udder health status could be developed. The classification of the udder health status could be done with a specifity of 100 % for the status healthy, 96.6 % for the status subclinical infected and 91.2 % for the status clinical infected.

Thesis (MSc (Microbiology))--Stellenbosch University, 2008. / Mastitis is considered to be the most costly disease affecting the dairy industry. Management strategies involve the extensive use of antibiotics to treat and prevent this disease. Prophylactic dosages of antibiotics used in mastitis control programmes could select for strains with resistance to antibiotics. In addition, a strong drive towards reducing antibiotic residues in animal food products has lead to research in finding alternative antimicrobial agents. Streptococcus macedonicus ST91KM, isolated from bulgarian goat yoghurt, produces the bacteriocin macedocin ST91KM with a narrow spectrum of activity against Grampositive bacteria. These include mastitis pathogens Streptococcus agalactiae, Streptococcus dysgalactiae, Streptococcus uberis, Staphylococcus aureus and Staphylococcus epidermidis as well as Lactobacillus sakei and Micrococcus varians. Macedocin ST91KM is, according to tricine-SDS PAGE, between 2.0 and 2.5 kDa in size. The activity of macedocin ST91KM remained unchanged after 2 h of incubation at pH 2.0 to 10.0 and 100 min at 100 °C. The peptide was inactivated after 20 min at 121 °C and when treated with pronase, pepsin and trypsin. Treatment with α-amylase had no effect on activity, suggesting that the mode of action does not depend on glycosylation. Precipitation with 60 % saturated ammonium sulphate, followed by Sep-Pak C18 separation recovered 43 % of macedocin ST91KM. Amplification of the genome of strain ST91KM with primers designed from the sequence of the macedocin prescursor gene (mcdA) produced two fragments (approximately 375 and 220 bp) instead of one fragment of 150 bp recorded for macedocin produced by S. macedonicus ACA-DC 198. Strain ACA-DC 198 was not available. However, the DNA fragment amplified from strain LMG 18488 (ACA-DC 206), genetically closely related to strain ACADC 198, revealed 99 % homology to the mcdA of S. macedonicus ACA-DC 198 (accession number DQ835394). Macedocin ST91KM may thus be a related bacteriocin described for S. macedonicus. The peptide adsorbed equally well (66 %) to L. sakei LMG13558 and insensitive cells, e.g. Enterococcus faecalis BFE 1071 and FAIR E92, and Streptococcus caprinus ATCC 700066. Optimal adsorption of macedocin ST91KM was recorded at 37 °C and 45 °C and at pH of 8 - 10. Addition of solvents decreased adsorption by 50%, suggesting that the receptors to which the bacteriocin binds have lipid moieties. The addition of MgCl2, KI and Na2CO3 completely prevented adsorption of macedocin ST91KM to the target cells, possibly due to competitive ion adsorption on the bacterial cell surface. The peptide has a bacteriocidal mode of action, resulting in lysis and the release of DNA and β-galactosidase. Atomic force microscopy of sensitive cells treated with macedocin ST91KM have shown deformation of the cell structure and developing of irregular surface areas. Antimicrobial susceptibility patterns were evaluated against eighteen mastitis pathogens. All isolates tested were resistant to methicillin and oxacillin, but had minimum inhibitory concentrations (MICs) falling in the intermediate and susceptible range against erythromycin. S. agalactiae and S. epidermidis had the highest sensitivity to macedocin ST91KM. A teat seal preparation containing macedocin ST91KM effectively released bacteriocin inhibiting the growth of the bacterial pathogen. Macedocin ST91KM could form the basis for an alternative dry cow therapy to prevent mastitis infections in dairy cows, as it is effective against pathogens that display resistance to conventional antibiotic therapy.

Peu d’études ont exploré l’utilisation du traitement prolongé pour les mammites cliniques et aucune ne portait sur l’utilisation du traitement prolongé à base de ceftiofur pour le traitement des mammites cliniques légères à modérées. L’objectif de cette étude était d’évaluer l’efficacité du traitement intra-mammaire prolongé à base de ceftiofur pour les mammites cliniques légères à modérées, en considérant toutes les bactéries responsables ou seulement Staphylococcus aureus (S. aureus) et les streptocoques. Des vaches laitières (n = 241) de 22 élevages du Québec et de l’Ontario ont été inclues. L’étude a été conçue comme un essai clinique à allocation aléatoire. Pour chaque cas de mammite clinique légère à modérée, 125 mg d’hydrochloride de ceftiofur (Spectramast® LC) a été administré par voie intra-mammaire une fois par jour pour 2 ou 8 jours. Le pourcentage de guérison clinique 21 jours après la fin du traitement était de 89% (n = 98/110 pour chaque groupe, p = 0,95). Les pourcentages de guérison bactériologique 21 jours après la fin du traitement pour les groupes 2 jours et 8 jours étaient 32% (n = 15/47) et 61% (n = 25/41) respectivement pour toutes les bactéries (p < 0,01); 64% (n = 9/14) et 82% (n = 9/11) respectivement pour les streptocoques (p = 0,50); et 0% (n = 0/20) et 47% (n = 9/19) pour S. aureus (p < 0,01). Il n'y avait pas de différence entre les 2 groupes pour les nouvelles infections intra-mammaires (p = 0,30). Le traitement prolongé à base de ceftiofur est un choix raisonnable pour le traitement des mammites cliniques légères à modérées, en particulier causées par S. aureus. / Little research has focused on extended therapy in lactating cows with clinical mastitis and none were with ceftiofur. The objective of the present study was to evaluate the efficacy of an intramammary extended ceftiofur treatment for mild to moderate clinical mastitis, and to determine whether it would increase cure rates when considering any bacteria or more particularly Staphylococcus aureus (S. aureus) and streptococci. Holstein dairy cows (n = 241) from 22 herds located in Quebec and Ontario, Canada, were included. The study was designed as a randomized clinical trial. For each case of mild to moderate clinical mastitis diagnosed, 125 mg of ceftiofur hydrochloride (Spectramast® LC) was administered intramammary once a day for 2 or 8 consecutive days. Clinical cure rate 21 days after the last treatment was identical in each group (89%; n = 98/110; p = 0.95). Bacteriological cure rates 21 days after the last treatment for the 2- and 8-day regimens were 32% (n = 15/47) and 61% (n = 25/41), respectively, for all bacteria (p < 0.01); 64% (n = 9/14) and 82% (n = 9/11), respectively, for streptococci (p = 0.50); and 0% (n = 0/20) and 47% (n = 9/19), respectively, for S. aureus (p < 0.01). There were no statistical differences between experimental groups for new intramammary infections. Extended therapy appears to be a reasonable choice for the treatment of mild to moderate clinical mastitis, particularly in cases where S. aureus is the pathogen involved.

En Amérique du Nord, Mycoplasma bovis est le plus pathogène des mycoplasmes retrouvés chez les bovins. Les principales maladies qu’on lui associe (maladies respiratoires, mammites, arthrites septiques et otites moyennes et/ou internes) constituent un défi à l’industrie laitière à cause de la difficulté à les traiter et à les prévenir par une vaccination. L’objectif principal de ce projet était d’étudier l’excrétion nasale et la réponse sérologique à M. bovis chez les génisses de remplacement, entre la naissance et 7 mois d’âge, dans 4 troupeaux laitiers au Québec. Quatre-vingt-trois paires mère/génisse provenant de 4 cohortes de bovins laitiers étaient prélevées mensuellement (génisses : 0 à 7 mois ; mères : 0, 1 et 5 mois après vêlage). Écouvillons nasaux et échantillons de lait étaient analysés par culture bactériologique et par immunofluorescence indirecte. Les anticorps circulants étaient détectés par le test ELISA. À la naissance, la prévalence sérologique des génisses était supérieure à celle des mères (P = 0,01). La transmission de M. bovis aux génisses par le lait et par l’excrétion nasale des mères était faible. L’âge moyen (jour) d’une génisse à sa 1ère excrétion nasale et sa 1ère séroconversion à M. bovis était loin de la période néonatale: 77,5 ± 11,2 (n = 22) et 96,8 ± 7,4 (n = 36) respectivement. Conclusion, les vaches adultes n’ont constitué qu’une voie mineure de transmission de M. bovis aux génisses, la principale voie de transmission était fort probablement le contact direct ou indirect avec d’autres génisses excrétrices nasales de M. bovis. / In North America, Mycoplasma bovis is the most pathogenic mycoplasma found in cattle. The main diseases associated with it (respiratory disease, mastitis, septic arthritis and otitis median and/or internal) are a challenge to the dairy industry because of the difficulty to treat them and to prevent them by vaccination. The principal objective of this project was to study nasal shedding and serological response to M. bovis in replacement heifers, between birth and 7 months of age, in four dairy herds in Quebec. Eighty three pairs cow/heifer in 4 cohorts of dairy cattle were sampled monthly (heifers: 0 to 7 months; cows: 0, 1 and 5 months after calving). Nasal swabs and milk samples were analyzed by bacteriological culture and by indirect immunofluorescence. Circulating antibodies were detected by ELISA test. At birth, the serologic prevalence of heifers was significantly higher than the serologic prevalence of cows (P = 0,01). Transmission of M. bovis to heifers in milk and nasal shedding from cows was low. The average age (days) of a heifer for first nasal shedding and first seroconversion to M. bovis was far from the neonatal period: 77,5 ± 11,2 days (n = 22) and 96,8 ± 7,4 days (n = 36) respectively. Conclusion, cows were only a minor route of transmission of M. bovis to heifers, the main route of transmission was most likely the direct or indirect contact with other heifers shedding M. bovis in their upper respiratory tract.

L’objectif de ce travail a été de caractériser la souche MPEC Escherichia coli P4. Une étude phylogénétique a montré qu’elle appartenait au groupe phylogénétique A de l’espèce E. coli et que son génome “core” se rapprochait de celui de la souche commensale non pathogène E. coli K12 MG1655 appartenant également au groupe A. Une recherche au sein de son génome de gènes codant différents facteurs de virulence connus chez les autres pathotypes de l’espèce E. coli a permis de détecter uniquement la présence du gène traT, codant un facteur de résistance au sérum. Un criblage de quinze loci d’ARNt connus pour accueillir fréquemment des îlots génomiques, effectué au sein de son génome, a révélé, pour sept d’entre eux la présence de telles structures. Le séquençage partiel ou complet des régions aval à ces sept loci a montré la présence systématique de séquences nucléotidiques différentes de celles présentes chez E. coli K12 MG1655. Si l’analyse du contenu de ces îlots n’a pas encore permis d’expliquer directement la virulence d’E. coli P4, leur mise en évidence est une première de ce type au sein du pathotype MPEC et laisse envisager la découverte d’autres régions génomiques spécifiques à ce pathotype, pouvant expliquer son tropisme et sa nature. Par ailleurs, afin d’évaluer le rôle d’E. coli P4 dans la caséinolyse élevée du lait observée lors d’une mammite bovine, une sécrétion apparemment constitutive de quatre protéases extracellulaires a été mise en évidence par zymographie caséines. L’activité caséinolytique de ces enzymes ne semble toutefois pas significative, laissant envisager plutôt un rôle dans la virulence de la souche / The objective of this work was to characterize the MPEC Escherichia coli P4 strain. A phylogenetic study showed that it belongs to the phylogenetic group A of the E. coli species and that its core genome is similar to the one of the commensal non-pathogenic E. coli K12 MG1655 strain which also belongs to the group A. A search in its genome of different genes encoding virulence factors known among other pathotypes of the E. coli species was done and only the traT gene, encoding a serum resistance factor, was detected. A screening of fifteen tRNA loci known for frequently hosting genomic islands, made in its genome, revealed for seven of them the presence of such structures. The partial or complete sequencing of the regions downstream from these seven loci showed the systematic presence of nucleotide sequences different from those present in E. coli K12 MG1655. If the content analysis of these islands does not yet explain the virulence of E. coli P4, their highlighting is the first of this kind in the pathotype MPEC and suggests the discovery of other genomic regions specific to this pathotype, which may explain its tropism and its nature. In addition, to assessing the role of E. coli P4 in milk caseinolysis observed during bovine mastitis, a constitutive secretion of four extracellular proteases was highlighted by casein zymography. However, the caseinolytic activity of these enzymes does not seem significant. This fact may suggest a role in virulence of the strain

A exploração dos bubalinos para a produção de leite e elaboração de seus derivados é uma atividade que tem crescido nos últimos anos no Brasil. Os búfalos apresentam problemas sanitários semelhantes aos bovinos. Objetivou-se neste estudo, desenvolver o isolamento e caracterização fenotípica e molecular dos principais microrganismos causadores de mastites subclínicas em búfalas (Bubalus bubalis) em quatro granjas leiteiras da região central do Estado de São Paulo, Brasil, através dos testes contagem de células somáticas (CCS), contagem padrão em placas (CPP), caracterização fenotípica e bioquímica dos microrganismos isolados e confirmação pela técnica de PCR, assim como também avaliar o perfil de sensibilidade bacteriana aos principais antimicrobianos para os patógenos mais frequentemente isolados em casos de mastite subclínica em bubalinos. Foram coletadas 20 amostras de leite de búfalas em lactação no ano de 2013, que apresentaram CCS acima de 200.000 cel/mL em controles realizados pelas fazendas. Tendo como base a Instrução Normativa Nº62 de 2011 do Ministério da Agricultura, Pecuária e Abastecimento (MAPA), que trata de padrões microbiológicos para leite bovino. Das 20 amostras analisadas neste trabalho, apenas 1 (5%) estava dentro do padrão estabelecido (1 x 104 UFC/mL) com uma variação nos resultados encontrados de 1,0 x 104 UFC/mL a 57,7 x 105 UFC/mL. A CCS apresentou uma mediana de 721.000 cel/mL no leite (valor mínimo: 205.000, valor máximo: 2.264.000), indicando presença de mastite subclínica. Constatou-se que os agentes com maior frequência de isolamento foram Staphylococcus epidermis (17%), Staphylococcus aureus (15%) e Bacillus spp. (14%). Os ensaios de PCR confirmaram os resultados obtidos com o isolamento na caracterização fenotípica e bioquímica de Staphylococcus aureus, Staphylococcus epidermis, Streptococcus spp. e Escherichia coli. Com a realização do antibiograma, foi possível traçar um perfil de sensibilidade aos diferentes antibióticos testados. O estudo de sensibilidade bacteriana detectou maior sensibilidade (100%) do Staphylococcus aureus e Staphylococcus epidermis aos antibióticos gentamicina e vancomicina; para o gênero Streptococcus spp. à gentamicina e oxacilina e para Escherichia coli à ampicilina. / The exploration of buffaloes for milk production and dairy products is an activity that has grown in recent years in Brazil. Buffaloes present similar health problems for cattle. The objective of this study was to determine the isolation and the phenotypic and molecular characterization of the main microorganisms causing of subclinical mastitis in buffaloes (Bubalus bubalis) on four dairy farms in the central region of state of Sao Paulo, Brazil, through testing somatic cell count (SCC), standard plate count (SPC), phenotypic and biochemical characterization of the microorganisms isolated and confirmed by PCR technique, as well as to evaluate the profile of bacterial sensitivity to antibiotics given for the major pathogens commonly isolated in cases of subclinical mastitis in buffaloes Twenty milk samples from lactating buffaloes were analyzed in 2013 that have presented SCC above 200,000 cells/mL. Based on the Instruction Normative N°62 of 2011 of the Ministério da Agricultura, Pecuária e Abastecimento (MAPA) which deals with microbiological standards for bovine milk, of the 20 samples analyzed in this study, only 1 (5%) was within the established standard (1x104 CFU/mL) with a variation in the results of 1.0x104 CFU/mL to 57.7x105 CFU/mL. The CCS showed a median of 721,000 cells/mL in the milk (minimum value; 205,000, maximum value: 2,264,000), indicating the presence of subclinical mastitis. It was found that agents with higher frequency of isolation were Staphylococcus epidermis (17%), Staphylococcus aureus (15%) and Bacillus spp. (14%). The PCR assays confirmed the results obtained with isolating in the phenotypic and biochemical characterization of Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus spp. and Escherichia coli. With the realization of the antibiogram was possible to draw a profile of resistance of different antibiotics tested. The study of bacterial sensitivity showed that for higher sensitivity (100%) of Staphylococcus aureus and Staphylococcus epidermidis to antibiotics gentamicin and vancomycin; for the genus Streptococcus spp. to gentamicin and oxacillin and for Escherichia coli to ampicilina.

Os objetivos do presente estudo foram: a) avaliar a frequência de isolamentos de patógenos causadores de mastite em rebanhos leiteiros comerciais; b) determinar a susceptibilidade antimicrobiana de Staphylococcus spp. e Streptococcus spp. isolados de casos de mastite subclínica c) avaliar o perfil de multirresistência de Staphylococcus spp. e Streptococcus spp. d) Detectar o gene mecA em Staphylococcus spp. resistentes a oxacilina/meticilina; e) avaliar a associação entre as práticas de manejo e tratamento de mastite e a susceptibilidade antimicrobiana de Staphylococcus aureus isolados de rebanhos leiteiros. Foram selecionados para o presente estudo 13 rebanhos leiteiros a partir de um total de 60 rebanhos vinculados a um laticínio da região de Pirassununga/SP. Questionários previamente formulados foram respondidos pelos responsáveis do rebanho para avaliar a associação entre as práticas de manejo e tratamento de mastite e a susceptibilidade antimicrobiana de S. aureus. Após a seleção dos rebanhos e aplicação dos questionários, 1069 amostras de leite compostas foram coletadas durante 24 meses, em quatro períodos para realização de cultura e identificação dos patógenos, testes de susceptibilidade antimicrobiana e detecção do gene mecA. Os testes de susceptibilidade foram realizados em todos os isolados de Staphylococcus spp. e em 50% de Streptococcus spp. selecionados aleatoriamente. Os antimicrobianos testados foram: ampicilina 10 mg; clindamicina 2 &micro;g, penicilina 1 mg; ceftiofour 30 &micro;g; gentamicina 10 mg; sulfatrimetropin 25 &micro;g, enrofloxacina 5 &micro;g; sulfonamida 300 &micro;g, tetraciclina 30 &micro;g; oxacilina 1 mg; cefalotina 30 &micro;g e eritromicina 5 &micro;g. Todos os isolados de Staphylococcus spp. que apresentaram resistência a oxacilina/meticilina foram avaliados quanto à presença do gene mecA. Entre os isolados, Staphylococcus coagulase negativa foi o mais prevalente (28,6%), seguido por S. aureus (27,8%) e Corynebacterium spp. (10,9%). A susceptibilidade antimicrobiana dos isolados de S. aureus foi de 95,23% (eritromicina), 93,33% (cefalotina) e 65,71% (ampicilina). A susceptibilidade antimicrobiana dos isolados de Staphylococcus coagulase negativa (SCN) foi de 93,33% (cefalotina), 89,91% (eritromicina) e 62,03% (ceftiofur). Entre os isolados de Staphylococcus coagulase positiva (SCP), a susceptibilidade antimicrobiana foi de 93,33% (eritromicina), 91,11% (clindamicina e sulfonamida). A susceptibilidade antimicrobiana dos isolados de Streptococcus agalactiae foi de 38,46% (ampicilina) e 50% (sulfatrimetropin) e os isolados de S. dysgalactiae apresentaram susceptibilidade à tetraciclina de 19,44% e a clindamicina de 47,22%. A multirresistência foi maior entre os isolados de SCN (20,3%) e S.aureus (15,7%). Não foi detectado o gene mecA em nenhum dos isolados de Staphylococcus spp. Houve associação entre as variáveis para os seguintes antimicrobianos: a) tratamento de mastite clínica: ampicilina (P = 0,0055), ceftiofur (P = 0,0481), sulfatrimetropin (P= 0,0293), sulfonamida (P = 0,0043) e tetraciclina (P = 0,0058); e envio de amostras para cultura e antibiograma: ampicilina (P=0,0032), tetraciclina (P=<0,0001). Os principais fatores de risco para susceptibilidade antimicrobiana de S. aureus foram o não envio de amostras para cultura e antibiograma para tetraciclina (OR=6.012), penicilina (OR=4.687), eritromicina (OR=3.059) e ampicilina (OR=2.571), tratamento de mastite clínica para ampicilina (OR=2.178), ceftiofur (OR=1.956), gentamicina (OR=1.668), oxacilina (OR= 1.132) e penicilina (OR= 1.261) e tratamento de vaca seca para enrofloxacina (OR=2.111) e tetraciclina (OR=2.075). Os microrganismos mais frequentemente isolados foram Staphylococcus coagulase negativa e S. aureus. Os testes de susceptibilidade realizados para as espécies de Staphylococcus spp. e Streptococcus spp. apresentaram alta susceptibilidade para a maioria dos antimicrobianos testados. Com exceção de Streptococcus agalactiae para ampicilina e sulfatrimetropin e os isolados de S. dysgalactiae para tetraciclina e clindamicina. A maioria dos Staphylococcus spp. e Streptococcus spp. não apresentaram perfil de multirresistência. Nenhum dos isolados de Staphylococcus spp. fenotipicamente resistentes a oxacilina/meticilina apresentaram o gene mecA. O maior fator de risco associado à susceptibilidade de S. aureus foi em relação à tetraciclina, penicilina, eritromicina e ampicilina para o não envio de amostras para cultura e antibiograma. Dois fatores de risco importantes identificados foram relacionados à ampicilina, ceftiofur, gentamicina, oxacilina e penicilina para o tratamento de mastite clínica e à enrofloxacina e tetraciclina para o tratamento de vaca seca. / The aims of this study were: a) to evaluate the frequency of mastitis pathogens in commercial dairy herds, b) to determine the antimicrobial susceptibility of Staphylococcus spp. and Streptococcus spp. isolated from subclinical mastitis c) to evaluate the profile of multidrug resistance of Staphylococcus spp. and Streptococcus spp. d) to detect the mecA gene in Staphylococcus spp. resistant to oxacillin / methicillin, e) to evaluate the association between management practices and treatment of mastitis, and antimicrobial susceptibility of Staphylococcus aureus isolates from dairy herds. Thirteen dairy herds were selected from a total of 60 dairy farms in the region of Pirassununga / SP. Questionnaires formulated previously were answered by dairy farmers to evaluate the association between mastitis treatment practices and antimicrobial susceptibility. A total of 1,069 composite samples of milk from all herds were collected for culture and identification of pathogens, susceptibility testing and mecA gene detection, over 24 months in four periods. The susceptibility tests were performed in all isolates of Staphylococcus spp. and in 50% of Streptococcus spp. randomly selected. The antimicrobials tested were ampicillin 10mg; 2&micro;g clindamycin, penicillin 1mg; ceftiofour 30&micro;g; gentamicin 10mg; sulfatrimetropin 25&micro;g, 5&micro;g enrofloxacin; 300&micro;g sulfonamide, tetracycline 30&micro;g; oxacillin 1mg; 30&micro;g cephalothin and erythromycin 5&micro;g. All Staphylococcus spp. that were resistant to oxacillin/methicillin in the susceptibility tests were investigated for the presence of mecA gene. Among the isolated bacteria, CNS was the most prevalent (28.6%), followed by S. aureus (27.8%) and Corynebacterium spp. (10.9%). The antimicrobial susceptibility of isolates of S. aureus was 95.23% (erythromycin), 93.33% (cephalothin) and 65.71% (ampicillin). The antimicrobial susceptibility of isolates of coagulase-negative Staphylococcus (CNS) was 93.33% (cephalothin), 89.91% (erythromycin) and 62.03% (ceftiofur). Among strains of coagulase positive Staphylococcus (CPS), the antimicrobial susceptibility was 93.33% (erythromycin), 91.11% (clindamycin and sulfonamide). The antimicrobial susceptibility of the strains of Streptococcus agalactiae was 38.46% (ampicillin) and 50% (sulfatrimetropin) and isolates of S. dysgalactiae were susceptible to tetracycline and clindamycin (19.44% to 47.22%). The multidrug resistance was higher among isolates of CNS (20.3%) and Staphylococcus aureus (15.7%). The mecA gene was not detected in any of Staphylococcus spp isolates. There was an association between variables for the following antimicrobials: a) treatment of clinical mastitis: ampicillin (P = 0.0055), ceftiofur (P = 0.0481), sulfatrimetropin (P = 0.0293), sulfonamide (P = 0.0043) and tetracycline (P = 0.0058); and sending samples for culture and antibiogram: ampicillin (P = 0.0032), tetracycline (P = <0.0001). Major risk factors for antimicrobial susceptibility of S. aureus were not submitting samples for culture and antibiogram for tetracycline (OR = 6.012), penicillin (OR = 4.687), erythromycin (OR = 3.059) and ampicillin (OR = 2.571), treatment of clinical mastitis: ampicillin (OR = 2.178), ceftiofur (OR = 1.956), gentamicin (OR = 1.668), oxacillin (OR = 1.132) and penicillin (OR = 1.261); and dry cow therapy: enrofloxacin (OR = 2.111) and tetracycline (OR = 2.075). The most frequent organisms isolated were coagulase negative Staphylococcus and S. aureus. The susceptibility tests performed for the species of Staphylococcus spp. and Streptococcus spp. showed high susceptibility to most antimicrobials tested. With the exception of Streptococcus agalactiae to ampicillin and sulfatrimetropin and isolates of S. dysgalactiae to tetracycline and clindamycin. Most Staphylococcus spp. and Streptococcus spp. did not show multidrug resistance profile. None of Staphylococcus spp. phenotypically resistant to oxacillin / methicillin showed the mecA gene. The major risk factor associated with susceptibility of S. aureus to tetracycline, penicillin, erythromycin and ampicillin was not sending samples for culture and sensitivity. Two important risk factors have been identified related to ampicillin, ceftiofur, gentamicin, penicillin and oxacillin for treatment of clinical mastitis and enrofloxacin and tetracycline for the treatment of dry cow dry cow therapy.

O objetivo deste trabalho foi verificar as possíveis correlações entre os índices de mastite e a qualidade microbiológica do leite cru, em 36 propriedades com atividade exploratória leiteira, no Estado de São Paulo, Brasil. Examinou-se 4662 quartos mamários de 1180 animais em lactação para se verificar a presença de mastite pelos Testes de Caneca de Fundo Escuro e CMT, e coletou-se uma amostra de cada quarto mamário positivo em pelo menos um dos testes para exame microbiológico. Para se avaliar a qualidade microbiológica do leite cru, coletou-se uma amostra de cada propriedade e fez-se a Contagem de microrganismos mesófilos aeróbios facultativos viáveis, Contagem de microrganismos psicrotróficos aeróbios facultativos viáveis, Contagem de microrganismos termófilos aeróbios facultativos viáveis, Contagem de Enterococcus spp., Contagem de Stapylococcus spp., Contagem de Streptococcus spp., Contagem de Corynebacterium spp., Contagem de bolores e leveduras, Número Mais Provável de coliformes totais e Número Mais Provável de coliformes fecais. Aplicou-se teste de Correlação de Pearson e Regressão Linear. Para comparação entre os índices de mastite, a melhor correlação foi entre o índice de resultados positivos ao teste CMT e os casos de mastite por causa infecciosa (r = 0,920 e R2 = 84,7%). Comparando-se o índice de mastite e a qualidade microbiológica do leite cru, o melhor resultado que apresentou correlação significativa foi entre o índice de casos de mastite por Staphylococcus spp. e a Contagem de microrganismos termófilos aeróbios facultativos viáveis (r = 0,522 e R2 = 27,3%). Pelo baixo número de análises que apresentaram resultado de Correlação significativa, notou-se que o índice mastite em rebanhos bovinos leiteiros não interfere na qualidade microbiológica do leite cru, nas condições estudadas neste experimento. / The purpose of this research was to investigate the possible correlations between the mastitis rate and raw milk microbiology quality, from 36 dairy farms, in São Paulo State, Brazil. It was investigated 4662 mammary quarters of 1180 lactant animals to examine mastitis cases using Tamis test and CMT. It was collected one sample of each positive mammary quarter in one test at least to microbiological analysis. To estimate the raw milk microbiological quality, it was colected one sample from each farm and realized one aerobic plate count of microorganisms mesophilic, aerobic plate count of microorganisms psychrotrophic, aerobic plate count of microorganisms termophilic, Enterococcus spp. plate count, Stahylococcus spp. plate count, Streptococcus spp. plate count, Corynebacterium spp. plate count, Yeast and Molds plate count, Most Probable Number of coliforms, Most Probable Number of fecal coliforms. It was used Pearson Correlation test and Linear Regression. In order to compare mastitis rates, the best correlation was between positives CMT test rate and the cases of mastitis caused by infections disease (r = 0,920 and R2 = 84,7%). Analysing the mastitis rate and the raw milk microbiological quality, the best result of correlation was between Staphylococcus mastitis rate and the aerobic plate count of microorganisms termophilic (r = 0,522 e R2 = 27,3%). Because of the low number of significant correlation, it was observed that the mastitis heard in dairy bovines herds do not interfere in raw milk microbiological quality, in the condition of this experiment.

O sistema de análise de perigos e pontos críticos de controle (APPCC) possui como principal objetivo a prevenção de riscos à saúde humana, bem como evitar alterações nos alimentos através da aplicação de práticas de controle em etapas da produção nas quais existe maior probabilidade de ocorrência de perigos ou situações críticas, sendo aplicado em todas as etapas de produção, desde a obtenção da matéria-prima até a elaboração do produto final. Desta maneira, o sistema APPCC é considerado como importante técnica para prevenção e controle de qualidade dos alimentos, já utilizado em fazendas leiteiras para controle de patógenos e resíduos de medicamentos. A contagem de células somáticas (CCS) e a contaminação bacteriana do Leite são fatores essenciais para manutenção da qualidade do leite em propriedades leiteiras, representando grandes prejuízos para a produção leiteira. Assim, os objetivos gerais da pesquisa foram: avaliação da eficiência, possíveis diferenças e principais problemas relacionados a aplicação do sistema APPCC em duas propriedades leiteiras (A e B) com vistas ao controle dos níveis de células somáticas e parâmetros higiênico-sanitários no leite do rebanho e ocorrência de mastite nos animais. Preliminarmente à implantação do sistema, realizou-se o treinamento em Boas Práticas de Produção e Manipulação para possibilitar a aplicação dos princípios do sistema APPCC. Os resultados mostraram que a aplicação do sistema foi eficaz para melhoria da qualidade do leite em uma das propriedades, resultado demonstrado pela melhoria da qualidade com referência ao controle da mastite e parâmetros higiênicos. Observou-se uma redução da CCS média dos animais, número de quartos afetados e escore de mastite. Quanto aos parâmetros higiênicos do leite, observou-se redução das contagens de mesófilos e coliformes fecais. Aspectos relacionados ao comprometimento dos funcionários e proprietário foram considerados essenciais para o sucesso do sistema APPCC, cuja implementação em propriedades leiteiras apresentou peculiaridades e diferenças pronunciadas em relação às indústrias alimentícias. / The Hazard Analysis Critical Control Point (HACCP) has, as the main objective the prevention of human’s health risks, as to avoid changes in the food due to use of the application of control practices in the steps of production, where the possibility of hazards or critical situations is bigger. The system is used in all steps of the production, since the ran material until the manufactured product. Regarding to this, the HACCP is considered as an important technique to prevent and to control the quality of the food that has been used in dainy forms to control pathogens and medical residues. The somatic cells count and the bacterial contamination of milk are essential factors to maintain the milk quality in the right condition, what represent big damages for the milk production. The objective of this research was to evaluate the efficiency, possible mistakes and the main problems related to the application of HACCP process in two dairy properties (A and B), based on the control of somatic cells level and hygienic conditions of the milk among the herd, besides the occurrence of the mastitis in animals. Before the application of the system, training in good manufacture practices and manipulation was accomplished to allow the use of the HACCP system. The results showed that the application of this method was efficient in order to improve the milk quality in one of the properties, what was shown in a better result of the mastitis’s control it was observed a reduction in the average counting of affected animals and mastitis’s cases. Related to the hygienic conditions of the milk, it was noticed a reduction in the numbers of mesophills and faecals coliforms. Aspects regarding the commitment of the workers and the owner were considered as essential for the success of the HACCP program, as the implementation in dairy forms showed particularities and differences related to the to the food industry.