Thesis (doctoral)--Universiteit Utrecht, 2003. / Includes bibliographical references (158-171).

Thesis (M.S.)--Ohio State University, 1985. / Includes bibliographical references (leaves 54-59). Available online via OhioLINK's ETD Center

This work is focused on the incidence of mastitis in three selected breeds with different reckoned milking. Breeds are a free stall and no litter technology. Resting boxes are lined with plastic litter. The authors compared the effects of milkings per day on the incidence of mastitis.

Tesis para optar al Grado de Magíster en Ciencias Animales y Veterinarias . / La mastitis es la principal enfermedad que afecta a vacas productoras de leche a nivel mundial, con costos de tratamiento de hasta US$ 200/vaca/año. Así mismo, las perdidas en producción de leche fluctúan entre 1.335 y 1.539 kg/lactancia/vaca en casos de mastitis sub clínica y clínica respectivamente. En Chile, dentro de los principales patógenos causantes de esta enfermedad se encuentra el Staphylococcus aureus (S. aureus) que es responsable del 22 al 68,6% de los cuadros sub clínicos. Recientemente, se ha reportado la existencia de un alto riesgo de resistencia bacteriana producto del uso inadecuado de antibacterianos para el tratamiento de mastitis causadas por S. aureus. Alternativamente en el último tiempo se ha establecido el uso de células madre mesenquimáticas (MSCs) para el tratamiento de múltiples enfermedades en medicina humana y veterinaria incluyendo infecciones bacterianas. En consecuencia, el objetivo del presente estudio fue evaluar el potencial antibacteriano in vitro contra S. aureus del medio condicionado de MSCs derivadas de médula ósea y tejido adiposo fetal bovino. Se utilizaron las cepas ATCC 25923 y SAU 1S de S. aureus con el fin de determinar su sobrevivencia en medio condicionado de MSCs no concentrado, concentrado, activado y activado concentrado mediante pre-exposición a S. aureus. En estos mismos cultivos de MSCs se cuantificaron los niveles de mRNA de los péptidos antimicrobianos β-defensina 4A (bBD4A), NK-lisina 1 (NK1), catelicidina 2 (CATHL2), hepcidina e indoleamina 2,3 dioxigenasa (IDO) mediante Q-PCR y de bBD4A mediante ELISA. La sobrevivencia de la cepa ATCC 25923 no fue diferente (P>0,05) cuando fue tratada con medio condicionado no concentrado de MSCs en comparación con los controles de fibroblastos y DMEM. Sin embargo, el porcentaje de sobrevivencia de la cepa SAU 1S disminuyó (P<0,05) al ser tratada con medio condicionado no concentrado de MSCs en comparación con el control DMEM. Adicionalmente, el medio condicionado concentrado de MSCs indujo una disminución (P<0,05) en la supervivencia de la cepa SAU 1S con respecto a los medios condicionados concentrados control de fibroblastos y DMEM. El medio condicionado activado, así como el medio condicionado activado concentrado de MSCs disminuyeron (P<0,05) la supervivencia de la cepa SAU-1S con respecto a los medios controles de MSCs no activados, de fibroblastos y DMEM. La activación indujo un aumento (P<0,01) de 3,5 veces en los niveles de mRNA de bBD4A en las MSCs de tejido adiposo y de 1,17 veces en MSCs de médula ósea. Además, la activación aumentó (P<0,05) en 1,66 y 1,37 veces el nivel de mRNA de NK1 en fibroblastos y MSCs de tejido adiposo, respectivamente. Los niveles de proteína bBD4A también aumentaron (P<0,05) en los medios condicionados de MSCs y de fibroblastos por efecto de la concentración. La activación solo aumenta (P<0,05) los niveles de bBD4A en los medios condicionados de MSCs derivadas de médula ósea. En conclusión, los medios condicionados de MSCs derivadas de médula ósea y tejido adiposo fetal bovino poseen efecto antibacteriano contra S. aureus en un sistema de cultivo in vitro. Este potencial antibacteriano se ve aumentado al concentrar por filtración y activar el medio condicionado mediante pre-exposición de MSCs a S. aureus. Estos resultados sugieren que el efecto antibacteriano de MSCs es mediado por la expresión de péptidos antimicrobianos NK1 y bBD4A. / Mastitis is the main disease affecting dairy cows worldwide with treatment costs reaching up to US $ 200/cow/year. Moreover, milk production losses fluctuate between 1,335 and 1,539 kg/lactation/cow in cases of clinical and clinical sub mastitis respectively. In Chile, one of the main pathogens causing mastitis Staphylococcus aureus (S. aureus), which is responsible for 22 to 68.6% of subclinical cases. Recently a high risk of bacterial resistance has been reported due to inappropriate use of antibacterial agents in the treatment of mastitis caused by S. aureus. Alternatively, the use of mesenchymal stem cells (MSCs) has been described for the treatment of multiple diseases in human and veterinary medicine including bacterial infections. Consequently, the aim of the present study was to evaluate the in vitro antibacterial potential against S. aureus of conditioned medium from MSCs derived from bone marrow and fetal bovine adipose tissue. S. aureus trains ATCC 25923 and SAU 1S were used in order to determine their survival in conditioned medium from non-concentrated, concentrated, activated and activated concentrated MSCs by exposure to S. aureus. mRNA levels from antibacterial peptides β-defensin 4A (bBD-4A), NK-lysine 1 (NK1), catelicidin 2 (CATHL2), hepcidin and Indoleamine 2,3 dioxygenase (IDO) were quantified by quantitative-PCR (Q-PCR) and bBD-4A by ELISA. The survival percentage of ATCC strain 25923 was not different (P>0.05) when it was treated with non-concentrated conditioned medium of MSCs compared to controls of fibroblasts and DMEM. In contrast, the survival percentage of SAU-1S strain decreased (P<0.05) when it was treated with nonconcentrated conditioned medium of MSCs compared to the DMEM control. Additionally, the concentrated conditioned medium of MSCs induced a decrease (P<0.05) in the survival percentage of the SAU 1S strain in comparison to the conditioned medium control fibroblasts and DMEM. The activated conditioned medium as well as the conditioned and activated conditioned medium of MSCs decreased (P<0.05) survival percentage of the SAU 1S strain with respect to non-activated MSCs control media, fibroblasts and DMEM. Activation induced a 3.5-fold increase (P<0.01) in bBD4A mRNA levels in adipose tissue MSCs and 1.17-fold in bone marrow MSCs. In addition, activation increased (P<0.05) 1.66 and 1.37-fold levels of NK1 mRNA in fibroblasts and adipose tissue MSCs, respectively. Levels of bBD4A also increased (P<0,05) in the conditioned media of MSCs and fibroblasts by concentration effect. Moreover, activation increased levels of bBD4A in conditioned media of MSCs derived from bone marrow. In conclusion, conditioned medium of MSCs possess antibacterial effect against S. aureus in an in vitro culture system. The antibacterial property is increased when conditioned medium is concentrated by filtration and activated by pre-exposition to S. aureus. The antibacterial potential of MSCs may be exerted by expression of antimicrobial peptides bBD4A and NK1. / Financiamiento: Proyecto Fondef ID 15I10129.

Made available in DSpace on 2014-06-11T19:27:16Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-11-30Bitstream added on 2014-06-13T20:16:20Z : No. of bitstreams: 1 nobrega_db_me_botfmvz.pdf: 839809 bytes, checksum: cf66e109eac48d5e6d7140e6607bfbf2 (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / O presente trabalho objetivou o isolamento de Klebsiella pneumoniae do ambiente de sala de ordenha e do ambiente dos animais, incluindo a sala de pré-ordenha, piquetes e, em algumas propriedades os galpões de free stall, do reto e membros posteriores dos animais, do leite do tanque de refrigeração e do leite dos tetos positivos ao teste do Califórnia Mastitis Test (CMT), pesquisar cepas produtoras de β-lactamases de espectro ampliado (ESβL) e, a partir da identificação clonal, verificar a semelhança entre as cepas. Em 28 (2,14%) amostras de leite coletadas dos animais foi detectada a presença de Klebsiella pneumoniae como patógeno responsável pela infecção intramamária (IMI). A partir de swabs ambientais, da pele dos animais e do leite oriundo do tanque de expansão das dez propriedades, foram isoladas 79 cepas de K. pneumoniae. Os resultados da prova de detecção fenotípica da produção de enzimas do tipo ESβL foram positivos em oito (7,47%) das cepas bacterianas isoladas. No Brasil ainda não há relatos de coliformes produtores destas enzimas oriundos de animais de produção, e no presente estudo uma das cepas foi isolada do tanque de expansão, representando um potencial perigo à saúde pública. Pelo resultado da PFGE, observou-se grande diversidade genética da bactéria dentro da propriedade e entre as propriedades. Demonstrou-se que patógenos ambientais multirresistentes estão presentes em rebanhos bovinos leiteiros, não apenas isolados de quadros de infecção intramamária como também de galpões de free stall, membros posteriores dos animais, sala de ordenha e tanque de expansão. Confirmou-se também, a alta variabilidade genética da K. pneumoniae em rebanhos bovinos leiteiros mesmo dentro de um mesmo rebanho, e a importância da PFGE como recurso na epidemiologia molecular / The present study aimed the isolation of Klebsiella pneumoniae from the milk parlor and the animal’s environment, including the pre milking room, paddocks and, in some properties the free stall, animal’s rectum and hind limbs, bulk tank milk and from all teats positive to the California Mastitis Test (CMT), search for strains producers of extended spectrum β-lactamases (ESβL) and, by the clonal identification, verify if similarities between the strains ocurred. In 2.14% (28) of the milk samples collected from the animals it was detected the presence of Klebsiella pneumoniae as the pathogen responsible for the intramammary infection (IMI). From environmental swabs and bulk milk tank of the ten properties, it was isolated 79 strains of K. pneumoniae. The results of phenotypic detection of the ESβL enzymes production were positive in eight (7.47%) bacterial strains isolated. It is important to know that, in Brazil there are no reports of coliforms producers of these enzymes isolated from livestock animals, and in the present study one of the strains was isolated from the bulk tank milk, representing a potential hazard to public health. The results of PFGE showed high genetic diversity of the bacteria within and between the dairy farms. It was demonstrated that environmental multidrug resistant pathogens are present in dairy herds, isolated from intramammary infections cases, free stall parlors, animal’s hindlimbs, milking parlor and bulk tank milk. This study also confirmed the high genetic diversity of K. pneumoniae in dairy herds within the same herd, and the importance of PFGE as a molecular epidemiology tool. Keywords: environment, extended-spectrum β-lactamases (ESβL), Klebsiella pneumoniae, mastitis, milk, pulsed-field gel electrophoresis (PFGE)

Made available in DSpace on 2016-08-12T18:48:53Z (GMT). No. of bitstreams: 0 Previous issue date: 2015-12-08. Added 1 bitstream(s) on 2016-08-12T18:51:05Z : No. of bitstreams: 1 000864826.pdf: 2067883 bytes, checksum: e9cd26ecf95e177b6e9775d4abb56c7d (MD5) / O estudo teve como objetivo avaliar as alterações da secreção láctea de cabras com mastite mediante o uso do teste da caneca de fundo escuro, California Mastitis Test (CMT), contagem de células somáticas (CCS) e perfil microbiológico, bem como determinar o perfil bioquímico, em especial de proteínas de fase aguda, do soro sanguíneo e soro lácteo de cabras com mastite clínica ou subclínica, de ocorrência natural. Foram constituídos três grupos experimentais compostos de 30 metades mamárias de cabras sadias, com secreção láctea negativa na prova da caneca de fundo escuro, CMT e no exame microbiológico (grupo controle - G1), 30 metades mamárias com secreção láctea negativa ao teste da caneca de fundo escuro, reação moderada (2+) ou intensa (3+) no CMT e positividade no exame microbiológico (Grupo mastite subclínica - G2) e 12 metades mamárias com secreção láctea positiva na prova da caneca de fundo escuro e no exame microbiológico (grupo mastite clínica - G3). Após antissepsia dos tetos foram colhidas amostras de leite para CCS, cultura microbiológica e determinação do perfil bioquímico do soro lácteo, inclusive o proteinograma mediante fracionamento em gel de poliacrilamida (SDS-PAGE), e amostras de sangue venoso para determinação do perfil bioquímico sérico. A contagem automática de células somáticas (CCS) e por contagem microscopia direta foi maior nas amostras de secreção láctea das cabras com mastite subclínica (G2: 4.500.000 células/mL e 20.830.000 células/mL, respectivamente) e com mastite clínica (G3: 7.500.000 células/mL e 39.100.000 células/mL, respectivamente). Staphylococcus aureus, Staphylococcus coagulase negativa (SCN), Staphylococcus coagulase positiva (SCP), Corynebacterium spp. e Streptococcus spp. foram os microrganismos isolados nas amostras de leite das cabras do G2 e G3. As amostras de soro lácteo das cabras do G2 e G3 apresentaram maiores atividades das enzimas... / The study aimed to evaluate the changes in milk from goats with mastitis by using the strip cup test, California Mastitis Test (CMT), somatic cell count (SCC) and microbiological profile as well as to determine the biochemical profile, especially acute phase proteins, in blood serum and whey from goats with naturally occurring clinical or subclinical mastitis. Three experimental groups were formed by 30 mammary glands from healthy goats with milk samples negative to strip cup test, CMT and microbiological examination (control group - G1); 30 mammary glands with milk samples negative to strip cup test but with moderate (2+) or intense (3+) reaction in CMT and microbiological examination (group with subclinical mastitis - G2), and 12 mammary glands with positive results to strip cup test and microbiological examination (group with clinical mastitis - G3). After asepsis of the teats milk samples were collected for SCC, microbiological culture, and to determine the biochemical profile in whey, including proteinogram by fractionation in poliacrylamide gel electrophoresis (SDS-PAGE). At the same time, blood samples were obtained to determine the serum biochemical profile. Automatic and microscopic methods were higher in milk samples from goats with subclinical mastits (G2: 4,500.000 cells/mL e 20,830,000 cells/mL, respectively) e with clinical mastitis (G3: 7,500,000 células/mL e 39,100,000 cells/mL, respectively). Staphylococcus aureus, Coagulase-negative Staphylococcus (CNS), Coagulase-positive Staphylococcus (CPS), Corynebaterium spp. and Streptococcus spp. were the microorganisms isolated in milk samples from goats of G2 and G3. Whey samples from goats of G2 and G3 showed higher activity of alkaline phosphatase (G2: 122 U/L and G3: 207 U/L) and gamma-glutamyltransferase (G2: 483 U/L and G3: 571 U / L), and higher concentrations of albumin (G2: 0.18 g/dL and G3: 0.29 g/dL), total protein (G2: 1.50 g/dL and G3: 1.67 g/dL), chlorides ...

Target is to explore, if homeopatics or another unconventional ways could be used in medical treatment or in prevention of cow mastitis, and at the same time to find by the mesurement of microclimatics indicators if the climate in stable has influence on inflammations of lacteal gland.

Tesis para optar al Grado de Magíster en Ciencias Animales y Veterinarias / La mastitis bovina es la enfermedad de mayor prevalencia en la industria lechera y la de mayor importancia, ya que provoca enormes pérdidas económicas que oscilan entre el 30 y el 70% de las pérdidas totales. El principal problema a nivel productivo está representado por la mastitis subclínica, la cual tiende a la persistencia o cronicidad, y a ser refractaria a tratamiento antibiótico. Este tipo de mastitis es comúnmente causado por Staphylococcus aureus y suele ser la forma de presentación más frecuente en los planteles lecheros a nivel mundial. El objetivo de esta tesis fue determinar diferencias entre cepas de S. aureus provenientes de vacas con mastitis persistentes y de vacas con mastitis transitorias respecto a su capacidad de formación de biofilm e invasión celular, y a su genotipificación capsular. Para ello, se seleccionaron aislados provenientes de mastitis bovinas persistentes y transitorias, que fueron caracterizados según su perfil de resistencia a antibióticos y tipificados mediante la técnica de Electroforesis en Campo Pulsado (Pulsed-Field). Se evaluó in vitro la formación de biofilm en placas de 96 pocillos y la invasión celular con células MAC-T. La genotipificación capsular (cap5-cap8) de las cepas se llevó a cabo mediante técnica de PCR. Además, mediante esta misma técnica se realizó la detección de los genes implicados en la formación de biofilm e invasión celular (operón icaADBC, bap, fnbA, fnbB). Se lograron identificar aislados de S. aureus persistentes y transitorios (n=30) en la glándula mamaria bovina mediante PFGE. Dichos aislados se agruparon en diferentes pulsotipos, demostrando una elevada diversidad genética, y en general fueron sensibles a todos los antibióticos ensayados (n=12), incluída la meticilina. El gen cap5 fue el de mayor frecuencia, pesquisándose en todas las cepas provenientes de vacas con mastitis persistentes y en una cepa proveniente de una vaca con mastitis transitoria; el resto de las cepas fueron “no tipificables”. Todas las cepas analizadas fueron capaces de formar biofilm, sin embargo, las cepas de S. aureus provenientes de vacas con mastitis persistentes formaron mayor cantidad de biofilm que las cepas de S. aureus provenientes de vacas con mastitis transitorias. En paralelo, todas las cepas fueron capaces de invadir células MAC-T, sin embargo, esta última capacidad fue baja en general y no se asoció con la manifestación de la mastitis o con el origen de las cepas (cepas provenientes de vacas con mastitis persistentes o mastitis transitorias). En conclusión, las cepas de S. aureus provenientes de vacas con mastitis persistentes codificaron en su genoma el gen cap5 y formaron mayor cantidad de biofilm que las cepas de S. aureus provenientes de vacas con mastitis transitorias / Bovine mastitis is the most prevalent and important disease in the dairy industry, since it causes huge economic losses ranging among 30 and 70% of total losses. The main problem at the production level is represented by subclinical mastitis, which tends to persistence or chronicity and to be refractory to antibiotic treatment. This type of mastitis is commonly caused by Staphylococcus aureus and is usually the most frequent form of presentation in dairy farms in the world. The aim of this thesis was to determine differences between strains of S. aureus from cows with persistent mastitis and cows with transient mastitis respect to their biofilm formation capacity, cell invasion capacity, and their capsular genotyping. isolates from persistent and transient bovine mastitis were selected and characterized according to their antibiotic resistance profile and typified by the Pulsed-Field technique. The formation of biofilm in 96 well plates and the cellular invasion with MAC-T cells were evaluated. Capsular genotyping (cap5-cap8) of the strains was carried out by PCR technique. In addition, the detection of the genes involved in the formation of biofilm and cellular invasion (operon icaADBC, bap, fnbA, fnbB) was carried out using the same technique. It was possible to identify persistent and transient S. aureus isolates (n=30) in the bovine mammary gland using the Pulsed-Field technique. Those isolates were grouped into different pulsotypes showing a high genetic diversity, and in general were sensitive to all antibiotics tested (n=12), including methicillin. The cap5 gene was the most frequent, being identified in all strains from cows with persistent mastitis and in one strain from a cow with transient mastitis; the rest of the strains were "non-typeable". All the strains were able to form biofilm, however, strains of S. aureus from cows with persistent mastitis formed more biofilm than strains of S. aureus from cows with transient mastitis. In parallel, all the strains were able to invade MAC-T cells, however, this last capacity was low in general and was not associated with the manifestation of mastitis or with the origin of the strains (strains from cows with persistent mastitis or transient mastitis). In conclusion, S. aureus strains from cows with persistent mastitis encoded the cap5 gene in their genome and formed more biofilm than S. aureus strains from cows with transient mastitis

No description available.

Three studies were conducted to assess if the ability of Enterococcus spp. surviving the ensiling process and bovine gastrointestinal tract could impact risk of bovine mastitis. The first study determined ability of enterococci to survive 3 wk ensiling. Grass and corn crops were divided into 3 treatments: 2 commercial silage inoculants, 1 negative control. After wk 1, 2, and 3 of ensiling, dry matter and bacterial enumeration were performed. Addition of silage inoculant led to greater levels of enterococci in grass silage compared with negative control levels, but showed less difference in inoculated corn silage. The second study quantified enterococci shedding rates in lactating dairy cows. Using a 4 x 4 Latin Square design, lactating, ruminally fistulated Holsteins were inoculated with enterococcal isolates from silage inoculants, ensiled forages, or clinical mastitis cases. Over the 4-period study, each period consisted of rumen and fecal sampling (2 wk) followed by a wash period (10 d). There were no significant differences in rumen or fecal enterococci levels between the 4 treatments. Both rumen and fecal enterococci levels showed significant differences between baseline and treatment periods (period 3, 4). The third study analyzed similarity in enterococcal isolates of silage and bovine origin using pulsed-field gel electrophoresis patterns from SmaI restrictions. Dendogram analysis showed none of the isolates met or were greater than a 75% genetic similarity and produced a genetically diverse population. Results suggest Enterococcus spp. from silage inoculants are not likely to contribute to an increased risk of enterococcal bovine mastitis. / Master of Science