ABSTRACT
Bovine mastitis causes economic losses on dairy farms worldwide. Lactic acid bacteria (LAB) in animal health are an alternative tool to avoid antibiotic therapy on the prevention of bovine mastitis. In previous studies, 12 LAB isolated from bovine milk were selected taking into account some of the following characteristics: hydrophobicity, auto aggregative capability, inhibition of indicator pathogens, hydrogen peroxide, and capsular polysaccharide production. These LAB were considered because of their beneficial properties. In this work, we also analyzed the antimicrobial activity and the co-aggregation against mastitis causing bacteria, auto-inhibition, adhesion to bovine teat canal epithelial cells (BTCEC), and growth kinetic curves for the 12 LAB. Two of them, Lactococcus lactis subsp. lactis CRL 1655 and Lactobacillus perolens CRL 1724, were selected because they had an interesting pattern of adhesion to BTEC, the inhibition of pathogens and the co-aggregation with the 100% of the assayed pathogens. They showed a predictable difference in the PFGE genomic pattern bands. The kinetic growth of these two strains was similar between them and with the rest of the assayed LAB. The strains selected in the present study showed indispensable characteristics for their inclusion in a probiotic formulation to be used at dry-off period for the prevention of bovine mastitis.
Subject(s)
Lactobacillales/physiology , Mastitis, Bovine/prevention & control , Milk/microbiology , Probiotics/pharmacology , Animals , Anti-Infective Agents/pharmacology , Bacterial Adhesion , Cattle , Epithelial Cells/microbiology , Female , Lactobacillales/genetics , Lactobacillales/isolation & purificationABSTRACT
Coagulase-negative staphylococci (CNS) are a common cause of bovine subclinical mastitis (SCM). The prevalence of CNS species causing SCM identified by genotyping varies among countries. Overall, the antimicrobial resistance in this group of organisms is increasing worldwide; however, little information exists about a CNS species resistant to antibiotics. The aim of the present study was to genotypically characterize CNS at species level and to determine the prevalence and antibiotic resistance profiles of CNS species isolated from bovine SCM in 51 dairy herds located in the central region of the province of Cordoba, Argentina. In this study, we identified 219 CNS isolates at species level by PCR-restriction fragment length polymorphism of the groEL gene. Staphylococcus chromogenes (46.6%) and Staphylococcus haemolyticus (32%) were the most prevalent species. A minimum of three different CNS species were present in 41.2% of the herds. S. chromogenes was isolated from most of the herds (86.3%), whereas S. haemolyticus was isolated from 66.7% of them. The broth microdilution method was used to test in vitro antimicrobial susceptibility. Resistance to a single compound or two related compounds was expressed in 43.8% of the isolates. S. chromogenes and S. haemolyticus showed a very high proportion of isolates resistant to penicillin. Resistance to two or more non-related antimicrobials was found in 30.6% of all CNS. S. haemolyticus exhibited a higher frequency of resistance to two or more non-related antimicrobials than S. chromogenes.
Subject(s)
Anti-Bacterial Agents/pharmacology , Staphylococcus/drug effects , Animals , Argentina , Cattle , Coagulase , Dairying , Female , Mastitis, Bovine/microbiology , Microbial Sensitivity Tests , Staphylococcus/enzymology , Staphylococcus/isolation & purificationABSTRACT
Coagulase-negative staphylococci (CNS) are a common cause of bovine subclinical mastitis (SCM). The prevalence of CNS species causing SCM identified by genotyping varies among countries. Overall, the antimicrobial resistance in this group of organisms is increasing worldwide; however, little information exists about a CNS species resistant to antibiotics. The aim of the present study was to genotypically characterize CNS at species level and to determine the prevalence and antibiotic resistance profiles of CNS species isolated from bovine SCM in 51 dairy herds located in the central region of the province of Cordoba, Argentina. In this study, we identified 219 CNS isolates at species level by PCR-restriction fragment length polymorphism of the groEL gene. Staphylococcus chromogenes (46.6%) and Staphylococcus haemolyticus (32%) were the most prevalent species. A minimum of three different CNS species were present in 41.2% of the herds. S. chromogenes was isolated from most of the herds (86.3%), whereas S. haemolyticus was isolated from 66.7% of them. The broth microdilution method was used to test in vitro antimicrobial susceptibility. Resistance to a single compound or two related compounds was expressed in 43.8% of the isolates. S. chromogenes and S. haemolyticus showed a very high proportion of isolates resistant to penicillin. Resistance to two or more non-related antimicrobials was found in 30.6% of all CNS. S. haemolyticus exhibited a higher frequency of resistance to two or more non-related antimicrobials than S. chromogenes.
Los estafilococos coagulasa negativos (ECN) son una causa frecuente de mastitis subclínica (MSC) en bovinos. La prevalencia de especies de ECN causantes de MSC identificadas por métodos genotípicos varía entre países. La resistencia antimicrobiana en este grupo de organismos se está incrementando en el mundo; sin embargo, existe poca información acerca de las especies de ECN resistentes a antibióticos. Los objetivos del presente estudio fueron caracterizar genotípicamente los ECN a nivel de especie y determinar la prevalencia y los perfiles de resistencia a antibióticos de las especies de ECN aisladas de MSC en bovinos de 51 rodeos situados en la provincia de Córdoba, Argentina. Mediante polimorfismos de los fragmentos de restricción del gen groEL identificamos 219 aislamientos de ECN a nivel de especie. Staphylococcus chromogenes (46,6%) y Staphylococcus haemolyticus (32%) fueron las especies más prevalentes. Un mínimo de 3 especies diferentes de ECN estuvieron presentes en el 41,2% de los tambos. S. chromogenes fue aislado en la mayoría de los tambos (86,3%), mientras que S. haemolyticus fue aislado en el 66,7% de aquellos. Para el análisis de sensibilidad a los antimicrobianos in vitro se usó el método de microdilución en caldo. La resistencia a un único compuesto o a 2 compuestos relacionados fue expresada en el 43,8% de los aislamientos. S. chromogenes y S. haemolyticus mostraron una muy elevada proporción de aislamientos resistentes a penicilina. La resistencia a 2 o más antimicrobianos no relacionados fue hallada en el 30,6% de los ECN. S. haemolyticus exhibió una frecuencia de resistencia a 2 o más antimicrobianos no relacionados más elevada que S. chromogenes.
Subject(s)
Animals , Cattle , Staphylococcus/drug effects , In Vitro Techniques/methods , Drug Resistance, Microbial/drug effects , Staphylococcus/classification , Mastitis, Bovine/drug therapyABSTRACT
The effect of intramammary inoculation of Lactobacillus perolens CRL 1724 on bovine udders at drying off was evaluated through histological examination of the canal and cistern tissues. The persistence of the strain in the udder 7 d post inoculation was also determined. Lb. perolens CRL 1724 was recovered from all mammary quarters and no clinical signs or teat damage were observed after inoculation of 10(6) cfu/ml. The udders showed a normal structural aspect and there were no modifications of the milk appearance. Lb. perolens CRL 1724 cells were evidenced on the surface of the epithelial cells of the cistern without causing any morphological modifications or cell alterations. Lb. perolens CRL 1724 produces a mild inflammatory reaction, characterized by recruitment of neutrophils to the epithelial zone and a slight hyperaemia into blood vessels. This preliminary study provides important information for further studies directed towards the inclusion of Lb. perolens CRL 1724 in the design of probiotic products for preventing bovine mastitis in non-lactating dairy cows.
Subject(s)
Cattle , Lactobacillus , Mammary Glands, Animal/anatomy & histology , Mammary Glands, Animal/microbiology , Animals , Epithelial Cells/microbiology , Epithelial Cells/ultrastructure , Female , Mastitis, Bovine/prevention & control , ProbioticsABSTRACT
Bovine mastitis is the most important infectious disease on dairy farms. Conventional antibiotic therapy is often unsatisfactory and alternative treatments are continually under investigation. Lactobacillus (Lb.) perolens CRL 1724 and Lactobacillus plantarum CRL 1716 were previously isolated from milk of dairy cows and selected according to their potential probiotic properties. In the present work the in-vitro capacity of Lactobacillus strains to adhere to bovine teat canal epithelial cells (BTCEC) and to inhibit and co-aggregate 14 mastitis-causing pathogens (MCPs) was investigated. The effect of Lb. perolens CRL 1724 after intramammary inoculation in lactating cows was evaluated through determination of clinical signs of mastitis, milk appearance, somatic cell counts and Lb. perolens CRL 1724 recovery from milk. Lb. perolens CRL 1724 was able to inhibit 12 of 14 MCPs (85·7%) in vitro, especially those considered to be major pathogens. In addition, Lb. perolens CRL 1724 co-aggregated with all of them. Lb. plantarum CRL 1716 was able to inhibit 7 of 14 MCPs (50%) in vitro and showed co-aggregation ability similar to Lb. perolens CRL 1724. Lb. perolens CRL 1724 showed a higher efficacy of adhesion to BTCEC (values of percentage of adhesion and adhesion index of 75% and 14·4, respectively) than Lb. plantarum CRL 1716 (37% and 7·4, respectively). Lb. perolens CRL 1724 was recovered from all mammary quarters and no clinical signs or teat damage were observed after the inoculation of 106 cfu/ml. The udders presented a normal aspect and there were no changes in the appearance of the milk. The results obtained will serve as the basis for further trials to evaluate the potential of Lb. perolens CRL 1724 to be included in a non-antibiotic formulation for the prevention of bovine mastitis.
