Information on the characteristics of Streptococcus agalactiae obtained from bovine sources in Brazil is still very limited. The aim of this study was to assess the phenotypic and genotypic diversity among S. agalactiae isolates from milk of dairy cows presenting clinical or subclinical mastitis in the southeast region of Brazil. Phenotypic characterization was based on physiological and serological tests. Antimicrobial susceptibility tests were carried out by the disk method. Genetic diversity was evaluated by using random amplified polymorphic DNA-PCR (RAPD-PCR) (by using the primer 1254) and pulsed-field gel electrophoresis (PFGE) (by using SmaI as the restriction enzyme) and by PCRs for detection of genes associated with resistance to erythromycin and tetracycline as well as PCRs for detection of genes coding for cell surface-associated proteins. According to the results of physiologic tests, 45 (52.9%) isolates showed beta-hemolysis and 44 (51.7%) were susceptible to bacitracin. Fourteen different biotypes were detected. The two most frequent biotypes comprised strains that were non-beta-hemolytic; fermented galactose, lactose, and salicin; produced protease; and were negative for DNase production. Serotype III was predominant (66 isolates [77.6%]), followed by serotypes II, Ia, Ib, and VI. Resistance to tetracycline and erythromycin was found in 38 (44.7%) and 9 (10.5%) isolates, respectively, with tet (O) (31.7%) and erm (B) (100%) being the most frequently occurring resistance genes. Three genes coding for surface proteins, bca , lmb , and scpB , were detected in 55 (64.7%), 7 (8.2%), and 43 (50.5%) isolates, respectively. In most cases, isolates from animals in the same herd presented closely related genetic profiles (determined by either RAPD-PCR or PFGE), which were distinct from those of isolates from different herds.
Bacteriocins have been suggested as an alternative to conventional antibiotics for the prevention and treatment of mastitis infections. Predominant bacteria associated with bovine mastitis (n = 276 isolates) were evaluated for their susceptibility to bovicin HC5, a ruminal bacteriocin produced by Streptococcus equinus HC5. Bovicin HC5 inhibited most (> 80%) of the streptococcal and staphylococcal strains tested, but showed no effect against Escherichia coli strains. Susceptibility and resistance testing indicated that approximately 95% of the S. aureus strains were inhibited by concentrations of bovicin HC5 varying from 40 to 2560 AU ml−1. Bovicin HC5 (62.50 AU ml−1) also inhibited the growth of aerobic and anaerobic mixed cultures of S. aureus and S. agalactiae, but the combination with 0.25 mmol l−1 of EDTA showed even greater bactericidal activity. These results demonstrate that bovicin HC5 is effective against the most prevalent pathogens found in contagious udder infections and could complement the use antibiotics in mastitis prophylaxis and therapy.
Bovine mastitis is mainly caused by bacteria of the genus Staphylococcus spp., which possess different virulence factors, including the capacity for biofilm formation that provides enhanced protection against the action of immune system components and serves as a barrier against the penetration of antimicrobial agents. This study aimed to characterize 181 Staphylococcus spp. Strains—including Staphylococcusaureus and coagulase-negative staphylococci (CoNS) isolated from bovine subclinical mastitis in six Brazilian states—by molecular methods. RT-qPCR was used to verify the expression of genes of the ica operon—mainly responsible for biofilm formation—as well as bap and bhp. Chromosome similarity among the isolates was investigated by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). The icaA gene was detected in 79 (43.6%) isolates, icaB in 24 (13.2%), icaC in 57 (31.4%), and icaD in 127 (70.1%). The bap gene was identified in 66 (36.4%) isolates, while the bhp gene was found in nine (4.9%). RT-qPCR confirmed the expression of the icaA gene in 60 (75.9%) isolates, of icaB in six (25%), of icaC in 26 (45.6%), and of icaD in 80 (63%). Clonal typing of the isolates by PFGE permitted the identification of eight Staphylococcusaureus clusters that simultaneously included ≥3 strains, with a similarity of ≥80%. Regarding the other species studied, three clusters were observed for Staphylococcuschromogenes and four clusters for Staphylococcusepidermidis. Only one cluster each was identified for Staphylococcussaprophyticus and Staphylococcussimulans, while the other species did not form any cluster. With respect to MLST, ST126 and ST1 were the prevalent sequence types in S. aureus, while in S.epidermidis all sequence types were different. These results reveal strains with the same evolutionary origin as other isolates, which might cause infections in humans and animals, suggesting their ability to spread between these species.
1 February 2007 in Arquivo Brasileiro de Medicina Veterinária e Zootecnia
The research was accomplished in eight dairy water buffalo herds, randomically choosen in Região do Alto São Francisco, State of Minas Gerais, Brazil. Information was collected from March to November, 2003 during 270 days of observation. In order to determine the somatic cell count (SCC) in presence or absence of microbial isolation, 1,393 samples were collected from 285 lactating females and microbiological exams and SCC were done. Samples obtained from udders without evidence of clinical or subclinical inflammation showed infection for a great variety of microbial mastitis pathogens. The low SCC did not necessarily indicate the absence of intramammary infection, suggesting that SCC patterns used for bovine cannot be appropriate in order to control mastitis in buffalo herds
1 November 2013 in Foodborne Pathogens and Disease
Minas Frescal cheese (MFC) is a traditional food produced and consumed in Brazil, characterized by its soft texture, low sodium, and high moisture content. This study characterized the microbiological contamination by coliforms, Escherichia coli and Staphylococcus aureus, in 99 MFC samples obtained in retail sale and produced by three distinct industrial procedures. Dairy processors were selected to investigate the key points of E. coli and S. aureus contamination during cheese processing. MFC samples produced by the addition of lactic culture presented higher counts of coliforms and E. coli, when compared to other samples (p<0.05). MFC samples produced by the addition of rennet alone presented higher counts of S. aureus when compared to other samples (p<0.05). Fourteen of 19 MFC samples produced by the addition of lactic culture presented E. coli counts higher than 5×102 colon-forming units/g. The processing steps after pasteurization were identified as the main sources of E. coli and S. aureus contamination of MFC. Based on the results, MFC was characterized as a potential hazard for consumers due to the high frequency of samples contaminated with E. coli and S. aureus counts in noncompliance with Brazilian standards for sanitary quality and safety.
1 January 2023 in Arquivo Brasileiro de Medicina Veterinária e Zootecnia
This study aimed to describe the epidemiological indexes of mastitis, milk quality and udder hygiene in the Compost Barn system, as well as to search for associations between isolated pathogens from milk with compost characteristics. Three dairy herds participated in the study, and the samples were collected during different periods on each farm. Individual milk samples were collected in duplicate for SCC analysis and microbiological culture. Environmental pathogens caused most cases of clinical mastitis on farm 2, and contagious pathogens caused the most cases on farm 1. Bed moisture was not associated with the incidence of environmental pathogens. Most of the animals remained in good udder hygiene during the study. Poor udder hygiene contributed to the increased incidence of environmental pathogens in one of the farms. A higher number of animals with a hygiene score of ≥ 2 were observed during the warmer and rainfall periods. There was no association between hygiene scores and somatic cell counts. The results suggest that pathogens isolated from milk in animals confined in Compost Barn under tropical climate are like other confinement systems adopted elsewhere. The year period influenced the udder hygiene score, reinforcing the importance of bed management throughout the year.