Subfertility in Buffaloes and the Association of Detected Milk Microbes

Lactogenesis is a complex physiology and an outcome of a successful reproductive cycle in mammalians. The purpose itself is to nourish the newborn with the nutrients necessary for its protection and growth a continuation from the in utero survival strategy. Thus, mammary development and physiology are intriguingly linked to the uterine changes that happen in a normal reproductive individual. Milk is a highly nutritious food that also favours the growth of microorganisms that can enter through the bloodstream or the discharge system. Briefly, the mastitis-induced release of inflammation mediators such as cytokines, interleukins, and prostaglandin F2? are established to be associated with infertility. Before breeding, mastitis can disrupt hormonal patterns (depression of estradiol production, delayed surge of luteinizing hormone) and delay ovulation. The issue of clinical mastitis is obvious but when subclinical mastitis persists for long the problem gets compounded. Thus, microbes get easily established in the mammary glands of buffaloes whose udder and teat anatomy, love for dirt, and swamp to wallow makes them more vulnerable exposing them to various kinds of pathogenic and opportunistic microbes. Buffaloes, by nature, have issues with silent heat, seasonal anestrus, more sensitive to direct radiation heat stress when faced with a pathological attack of infectious microbes in the milk chamber compromises their fertility.

Antimicrobial properties of heterocyclic compounds against clinical mastitis isolates

ABSTRACT: Mastitis causes significant economic losses to the dairy cattle industry. The present study aimed to evaluate the antibacterial properties of 39 heterocyclic derivatives (1,3-thiazoles and 4-thiazolidinones) against clinical mastitis isolates from dairy cows. Milk samples were collected from cows with clinical mastitis and the bacterial species were identified by PCR. Antibacterial activity was assessed using the broth microdilution method. First, 39 heterocyclic compounds were tested against four bacterial isolates (Staphylococcus aureus, Streptococcus agalactiae, Corynebacterium bovis and Escherichia coli) randomly chosen from those recovered from the milk samples (Study 1). Subsequently, the compounds with the strongest antibacterial activity were tested against all the bacterial isolates recovered from the milk samples (Study 2). 1,3-thiazoles showed the strongest antibacterial activity, specially compounds 30 and 38, which also showed bactericidal properties according to their minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) values. Corynebacterium spp. and Enterobacteriaceae isolates were the most susceptible to compounds 30 and 38. Compounds 30 and 38 are promising targets for new antimicrobial agents.

RESUMO: A mastite causa significativas perdas econômicas à indústria leiteira bovina. O presente estudo teve como objetivo avaliar as propriedades antibacterianas de 39 derivados heterocíclicos (1,3-tiazóis e 4-tiazolidinonas) contra isolados clínicos de mastite em vacas leiteiras. Amostras de leite foram coletadas de vacas com mastite clínica e as espécies bacterianas isoladas foram identificadas por PCR. A atividade antibacteriana foi avaliada pelo método de microdiluição em caldo. Primeiramente, os 39 compostos heterocíclicos foram testados contra quatro isolados bacterianos (Staphylococcus aureus, Streptococcus agalactiae, Corynebacterium bovis e Escherichia coli) escolhidos aleatoriamente dentre os recuperados das amostras de leite (Estudo 1). Posteriormente, compostos com atividade antibacteriana mais forte foram testados contra todos os isolados bacterianos recuperados das amostras de leite (Estudo 2). Os compostos 1,3-tiazóis apresentaram a maior atividade antibacteriana, principalmente os compostos 30 e 38, que também apresentaram propriedades bactericidas de acordo com seus valores de concentração inibitória mínima (CIM) e concentração bactericida mínima (CBM). Os isolados Corynebacterium spp. e Enterobacteriaceae foram os mais suscetíveis aos compostos 30 e 38. Os compostos 30 e 38 mostraram-se promissores como novos agentes antimicrobianos.

Antimicrobial properties of heterocyclic compounds against clinical mastitis isolates / Propriedades antimicrobianas de compostos heterocíclicos contra isolados de mastite clínica

Mastitis causes significant economic losses to the dairy cattle industry. The present study aimed to evaluate the antibacterial properties of 39 heterocyclic derivatives (1,3-thiazoles and 4-thiazolidinones) against clinical mastitis isolates from dairy cows. Milk samples were collected from cows with clinical mastitis and the bacterial species were identified by PCR. Antibacterial activity was assessed using the broth microdilution method. First, 39 heterocyclic compounds were tested against four bacterial isolates (Staphylococcus aureus, Streptococcus agalactiae, Corynebacterium bovis and Escherichia coli) randomly chosen from those recovered from the milk samples (Study 1). Subsequently, the compounds with the strongest antibacterial activity were tested against all the bacterial isolates recovered from the milk samples (Study 2). 1,3-thiazoles showed the strongest antibacterial activity, specially compounds 30 and 38, which also showed bactericidal properties according to their minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) values. Corynebacterium spp. and Enterobacteriaceae isolates were the most susceptible to compounds 30 and 38. Compounds 30 and 38 are promising targets for new antimicrobial agents.(AU)

A mastite causa significativas perdas econômicas à indústria leiteira bovina. O presente estudo teve como objetivo avaliar as propriedades antibacterianas de 39 derivados heterocíclicos (1,3-tiazóis e 4-tiazolidinonas) contra isolados clínicos de mastite em vacas leiteiras. Amostras de leite foram coletadas de vacas com mastite clínica e as espécies bacterianas isoladas foram identificadas por PCR. A atividade antibacteriana foi avaliada pelo método de microdiluição em caldo. Primeiramente, os 39 compostos heterocíclicos foram testados contra quatro isolados bacterianos (Staphylococcus aureus, Streptococcus agalactiae, Corynebacterium bovis e Escherichia coli) escolhidos aleatoriamente dentre os recuperados das amostras de leite (Estudo 1). Posteriormente, compostos com atividade antibacteriana mais forte foram testados contra todos os isolados bacterianos recuperados das amostras de leite (Estudo 2). Os compostos 1,3-tiazóis apresentaram a maior atividade antibacteriana, principalmente os compostos 30 e 38, que também apresentaram propriedades bactericidas de acordo com seus valores de concentração inibitória mínima (CIM) e concentração bactericida mínima (CBM). Os isolados Corynebacterium spp. e Enterobacteriaceae foram os mais suscetíveis aos compostos 30 e 38. Os compostos 30 e 38 mostraram-se promissores como novos agentes antimicrobianos.(AU)

Expression of L-Selectin in Peripheral Blood Leukocytes of Crossbred Cows Affected with Subclinical Mastitis

L-Selectin (SELL) linked with innate immune mechanism involved in neutrophil migration through endothelium to the site of inflammation. Early recruitment of neutrophils at the site of infection is essential to counteract infection at the earliest in conditions like sub-clinical mastitis (SCM). The present study was framed to evaluate the expression pattern of SELL in naturally occurring SCM in crossbred animals using real time PCR technique. Analysis of data on total leukocyte count indicated leucocytosis condition in SCM affected crossbred cows. Relative expression of SELL on peripheral blood leukocytes revealed a significant 3.16 folds down regulation in SCM affected cows when compared with healthy crossbred cows (P<0.05). The possible reason for leukocytosis in SCM animals might be due to down regulation of SELL on leukocytes reducing their ability to transmigrate through blood vessel to infection site. The result of present study revealed a definite role of SELL in SCM which could be explored for therapeutic aspects in near future

Incidence and Factors Affecting Subclinical and Clinical Mastitis in Selected Organised Dairy Farms Located in Tamil Nadu

The study is undertaken in five organised dairy farms located in Chennai, Trichy and Karur districts of Tamil Nadu to assess the incidence of clinical and sub clinical mastitis and predisposing factors such as Herd, Breed, Parity, stage of lactation and the efficiency of two screening tests for the diagnosis of sub clinical mastitis (SCM) in the dairy farms. A total of 159 cows with 636 quarters samples screened for clinical mastitis signs and SCM by modified California mastitis test (mCMT) and somatic cell count (SCC).The results showed that 9.4 per cent of animals had clinical mastitis, 66.7 per cent had SCM by mCMT and 77.4 per cent by SCC. The quarter wise incidence of SCM was 47.6 per cent as screened by mCMT and 51.7 per cent by SCC. There are no significant differences observed in the incidence of clinical mastitis and SCM in farm and breed wise. Parity (p<0.01), Stage of lactation by SCC method (p<0.05), Quarter-wise (p<0.05) incidence have significant differences of occurrence. SCM as screened by mCMT and SCC were compared for agreement by Cohen’s Kappa statistics and the per cent agreement for the two methods of screening SCM was 54.7 per cent with a P-value of 0.000. The study conclude that the higher SCM incidence in organised dairy farms as 66.7 per cent by mCMT and 77.4 per cent by SCC and necessitate the need of effective mastitis control program to be implemented at the farm level

Tasa de incidencia de mastitis clínica y susceptibilidad antibiótica de patógenos productores de mastitis en ganado lechero del norte de Antioquia, Colombia* / Incidence rate of clinical mastitis and antibiotic susceptibility of mastitis-producing pathogens in dairy cattle from Northern Antioquia, Colombia / Taxa de incidência de mastite clínica e susceptibilidade antibiótica de patógenos produtores de mastite em gado leiteiro do norte de Antioquia, Colômbia

Resumen Introducción: la mastitis se define como la inflamación de la glándula mamaria causante de grandes pérdidas económicas en hatos lecheros, lo que amerita su investigación. Objetivo: determinar la tasa de incidencia de mastitis clínica TIMC, la etiología asociada y la sensibilidad antibiótica de patógenos aislados de casos de mastitis en ganado lechero. Materiales y métodos: se siguieron 37 hatos por 12 meses para el diagnóstico de mastitis cílínica (MC). A las muestras de leche de los casos de MC se les realizó cultivo bacteriológico y a estas, así como a los aislamientos de casos de mastitis subclínica de un estudio previo, se les efectuó prueba de susceptibilidad antibiótica. Para el análisis de la información se utilizó estadística descriptiva estándar. Resultados: TIMC fue de 13,8 casos por 100 vacas-año a riesgo. Se aislaron 188 patógenos totales de casos de MC, entre los cuales el Streptococcus agalactiae, el Streptococcus pyogenes y el Corynebacterium spp. fueron los más frecuentes, con 29,8; 11,7 y 5,9 %, respectivamente. Se halló una alta sensibilidad de las bacterias contagiosas para los antibióticos cloxacilina y cefoperazone. Conclusiones: en el presente estudio se encontró una TIMC de 13,8 casos por 100 vacas-año a riesgo. Los patógenos más prevalentes identificados en casos de MC fueron contagiosos. Se encontró una alta sensibilidad de las bacterias contagiosas para la mayoría de los antibióticos β-lactámicos.

Abstract Introduction: Mastitis is defined as the inflammation of the mammary gland causing great economic losses in dairy herds, which merits investigation. Objective: To determine the incidence rate of clinical mastitis IRCM, associated etiology, and antibiotic sensitivity of pathogens isolated from cases of mastitis in dairy cattle. Materials and methods: 37 herds were followed during 12 months for clinical mastitis (CM) diagnosis. Milk samples from CM cases were bacteriologically cultured, which underwent, along with isolates from subclinical mastitis cases from a previous study, antibiotic susceptibility testing. For data analysis, standard descriptive statistics were used. Results: The IRCM was 13.8 cases per 100 cows-year at risk. A total of 188 pathogens of CM cases were isolated, among which Streptococcus agalactiae, Streptococcus pyogenes, and Corynebacterium spp. were the most frequent, with 29.8%, 11.7%, and 5.9%, respectively. A high susceptibility of contagious bacteria was found for cloxacillin and cefoperazone. Conclusions: The present study found an IRCM of 13.8 cases per 100 cows-year at risk. The most prevalent pathogens identified in cases of CM were contagious. A high sensitivity of contagious bacteria was found for most β-lactam antibiotics.

Resumo Introdução: a mastite se define como a inflamação da glândula mamária causante de grandes perdas econômicas em rebanhos de gado leiteiro, o que justifica a sua pesquisa. Objetivo: determinar a taxa de incidências de mastite clínica TIMC, a etiologia associada e a sensibilidade antibiótica de patógenos isolados de casos de mastite em gado leiteiro. Materiais e métodos: seguiram-se 37 rebanhos bovinos durante 12 meses para o diagnóstico de mastitis clínica (MC). Com as amostras de leite dos casos de MC realizou-se cultivo bacteriológico e a estas, assim como aos isolamentos de casos de mastite subclínica de um estudo prévio, efetuou-se prova de susceptibilidade antibiótica. Para a análise da informação se utilizou estatística descritiva padrão. Resultados: a TIMC foi de 13,8 casos por 100 vacas-ano a risco. Foram isolados 188 patógenos totais de casos de MC, entre os quais o Streptococcus agalactiae, o Streptococcus pyogenes e o Corynebacterium spp. foram os mais frequentes, com 29,8; 11,7 e 5,9 %, respectivamente. Constatou-se uma alta sensibilidade das bactérias contagiosas para os antibióticos cloxacilina e cefoperazone. Conclusões: neste estudo evidenciou-se uma TIMC de 13,8 casos por 100 vacas-ano a risco. Os patógenos mais prevalentes identificados em casos de MC foram contagiosos. Encontrou-se uma alta sensibilidade das bactérias contagiosas para a maioria dos antibióticos β-lactâmicos.

Frecuencia y susceptibilidad antimicrobiana del estafilococo coagulasa negativo aislado de mastitis bovina en fincas lecheras del Tolima, Colombia / Frequency and Antimicrobial Susceptibility of Coagulase-negative Staphylococci Isolated from Bovine Mastitis in Dairy Farms from Tolima, Colombia / Frequência e susceptibilidade antimicrobiana do estafilococo coagulase negativo isolado de mastite bovina em fazendas produtoras de leite do Tolima, na Colômbia

Objetivo: Determinar la presencia y susceptibilidad antimicrobiana del grupo estafilococo coagulasa negativo (ECN) en algunas fincas ganaderas de Tolima, Colombia. Materiales y métodos: Mediante la prueba de California para el diagnóstico de mastitis (CMT), se evaluaron 484 cuartos pertenecientes a 121 vacas de cinco fincas ganaderas de pequeña producción de una región del centro de Tolima. Las muestras positivas al CMT se cultivaron para aislamiento bacteriológico. A los ECN encontrados se les realizaron pruebas de susceptibilidad frente a los antibióticos. Los resultados se analizaron con estadística descriptiva. Resultados: 252 (52%) cuartos no presentaron ningún tipo de reacción al CMT, ni signos clínicos de mastitis, por lo cual se consideraron libres de la enfermedad. De los cuartos, 73 (15%) fueron positivos para CMT y cultivo bacteriológico. De estos se aislaron 36 cepas de ECN (7,4% del total de cuartos); S. aureus-ECP, 28 (5,8%); Streptococcus spp., 6 ( 1,2%); Escherichia coli, 2 (0,4%), y Klebsiella pneumoniae, 2 (0,2%). La mastitis clínica y subclínica en el cuarto se presentó en 1,4% y 13,6%, respectivamente. En 5 (1,0%) de los cuartos se encontró mastitis clínica causada por ECN y mastitis subclínica en 31 (6,4%). El 61% de las cepas de ECN fue resistente a penicilina, y el 58%, a tetraciclina; el 97% fue sensible a cefoperazona. Conclusión: El grupo ECN, considerado un emergente mundial de mastitis, es evidenciado con alta frecuencia en fincas ganaderas de Tolima, Colombia, lo que causa mastitis clínica y subclínica con variada respuesta a los antimicrobianos.

Objective: To determine the antimicrobial presence and susceptibility of the coagulase-negative staphylococci group (ECN, for its initials in Spanish) in some cattle farms from Tolima, Colombia. Materials and methods: Using the California test for the diagnosis of mastitis (CMT, for its initials in Spanish), 484 quarters belonging to 121 cows from five small production ranches from a region of central Tolima were evaluated. CMT positive samples were cultivated for bacterial isolation. The ECN found were tested for susceptibility to the antibiotics. The results were analyzed with descriptive statistics. Results: 252 (52%) quarters did not show any type of reaction to the CMT, nor did they show any clinical signs of mastitis, therefore they were considered free of the disease. From the quarters, 73 (15%) turned positive for CMT and bacteriological culture. From these, 36 strains of ECN were isolated (7.4% of the total of quarters); S. aureus-ECP, 28 (5.8%); Streptococcus spp., 6 (1.2%).; Escherichia coli, 2 (0.4%), and Klebsiella pneumoniae, 2 (0.2%). The clinical and subclinical mastitis in the quarter occurred in 1.4% and 13.6%, respectively. In 5 (1.0%) of the quarters, clinical mastitis caused by ECN was found and subclinical mastitis in 31 (6.4%). 61% of the ECN strains were resistant to penicillin, and 58%, to tetracycline; 97% were sensitive to cefoperazone. Conclusion: The ECN group, considered a global emergent of mastitis, is evidenced with high frequency in ranches from Tolima, Colombia, causing clinical and sub-clinical mastitis with varied response to antimicrobials.

Objetivo: Determinar a presença e susceptibilidade antimicrobiana do grupo estafilococo coagulase negativo (ECN) em algumas fazendas de gado do Tolima, na Colômbia. Materiais e métodos: Mediante a prova de Califórnia para o diagnóstico de mastite (CMT), se avaliaram 484 quartos pertencentes a 121 vacas de cinco fazendas de gado de pequena produção de uma região do centro do Tolima. As amostras positivas ao CMT se cultivaram para isolamento bacteriológico. Aos ECN encontrados foram-lhes realizadas provas de susceptibilidade diante dos antibióticos. Os resultados se analisaram com estatística descritiva. Resultados: 252 (52%) quartos não apresentaram nenhum tipo de reação ao CMT, nem signos clínicos de mastite, razão pela qual foram considerados livres da doença. Dos quartos, 73 (15%) foram positivos para CMT e cultivo bacteriológico. Destes se isolaram 36 cepas de ECN (7,4% do total de quartos); S. aureus-ECP, 28 (5,8%); Streptococcus spp., 6 (1,2%); Escherichia coli, 2 (0,4%), e Klebsiella pneumoniae, 2 (0,2%). A mastite clínica e subclínica no quarto se apresentaram em 1,4% e 13,6%, respectivamente. Em 5 (1,0%) dos quartos se encontrou mastite clínica causada por ECN e mastite subclínica em 31 (6,4%). Em 61% das cepas de ECN foram resistentes à penicilina, e o 58%, à tetraciclina; o 97% foram sensíveis à cefoperazona. Conclusão: o grupo ECN, considerado um emergente mundial de mastite, é evidenciado com alta frequência em fazendas de gado do Tolima, na Colômbia, o que causa mastite clínica e subclínica com variada resposta aos antimicrobianos.

Classification of acute clinical mastitis on the base of vital signs and complete blood count test in dairy cows / 대한수의학회지

This study was carried out to evaluate the diagnosis of acute clinical mastitis (ACM) which was based on the vital signs and complete blood count (CBC) tests in dairy cows. Twenty eight dairy cows diagnosed with ACM, were selected for the study between Jan 2003 and July 2006 in the National Institute of Animal Science. Based on their vital signs (rectal temperature, depression, rumen contraction and, dehydration status), ACM was divided into three different classes; mild, moderate and severe forms. In addition, ACM cows were subjected to CBC tests for further diagnosis of ACM. Of the 27 dairy cows diagnosed with ACM, 3 cows were determined to have a mild form, while moderate and sever forms were each observed in twelve cows. Among of them, 4 cows died, 5 cows were culled and 18 cows were recovered. In the mild form, all haematological parameters were comparable with normal values. However, leukopenia, due to neutropenia and lymphocytopenia, appeared characteristically in the moderate and severe forms. Using the observation of vital signs in conjunction with CBC tests, the diagnosis of ACM is more accurate, and is helpful in making decisions of whether treatment or culling of dairy cows infected with ACM is most appropriate.

New method for early detection of two random amplified polymorphic DNA (RAPD) groups of Staphylococcus aureus causing bovine mastitis infection in Paraná State, Brazil

The aim of this work was to develop a fast and accurate molecular approach to allow early detection of two RAPD groups of S. aureus causing bovine mastitis. Seventy five S. aureus isolates from infected animals were characterized by RAPD. Genomic fragments isolated from the unique bands present in either group were cloned and sequenced. Based on the DNA sequences, specific primers were designed to allow for the simultaneous detection of either group by multiplex PCR of S. aureus DNA isolated from clinical and subclinical bovine mastitis. Results showed that these proposed primers set could be used to detect various clinical and subclinical S. aureus isolates as well as the detection of the microorganism in bulk milk. Their use as a specific method for effective and early diagnostic tool for S. aureus infection in dairy herds is suggested.

Esta pesquisa objetivou o desenvolvimento de técnica rápida e eficiente para diagnosticar precocemente diferentes linhagens de S. aureus causadoras de mastite bovina. Como resultados da metodologia empregada, foram isoladas duas linhagens destas bactérias que causam diferentes tipos de mastite bovina. Os fragmentos de DNA genômico caracterizando ambas as linhagens, por meio de RAPD foram inseridos em vetor plasmidial pGEM e clonados por meio de clones T10 F1 de Escherichia coli. As seqüências obtidas permitiram desenhar iniciadores específicos para o reconhecimento de ambas as linhagens, os quais foram testados com amostras de S. aureus e com outras linhagens próximas. O diagnóstico por meios moleculares, pode ser realizado diretamente de amostras coletadas de rebanhos leiteiros assim como dos equipamentos de ordenha. A significância deste estudo consiste em um rápido e acurado método para localizar animais infectados, representando importante ferramenta no manejo do rebanho, na redução de custos com tratamentos e, rápida recuperação de rebanhos infectados.