Activity of staphylococcal bacteriocins against Staphylococcus aureus and Streptococcus agalactiae involved in bovine mastitis (original) (raw)

Bacteriocin production by Staphylococcus aureus involved in bovine mastitis in Brazil

Research in Microbiology, 2009

In the present study, 257 Staphylococcus spp. strains were isolated from bovine mastitis cases in 56 different Brazilian dairy herds located in the southeast region of the country and tested for antimicrobial substance (AMS) production. Forty-six strains (17.9%) exhibited AMS production and their identification as Staphylococcus aureus was based on the presence of Gram-positive cocci and on positive results in tests for the ability to coagulate rabbit plasma, to ferment mannitol, and to produce acetoin. The AMS were characterized as bacteriocins (Bac) by their sensitivity to proteolytic enzymes. The Bac þ strains were tested for resistance to 14 antimicrobial agents showing different profiles. Eighteen strains (39.0%) expressed a multiple antibiotic resistance phenotype. Forty-five strains exhibited at least one plasmid DNA. Cross-immunity analysis against strain S. aureus A70, which produces aureocin A70, amplification of the aurABCD operon (which encodes aureocin A70) or detection of this same operon by DNA/DNA hybridization revealed that 34 strains produce bacteriocins either identical or similar to aureocin A70. The remaining 12 Bac þ strains produce antimicrobial peptides that seem to be distinct from the best characterized staphylococcal bacteriocins described thus far. The bacteriocin produced by strain 4185 may possess potential practical applications, since it was able to inhibit important pathogens such as Bacillus cereus, Listeria monocytogenes, and Staphylococcus spp. isolated from nosocomial infections.

Production of bacteriocins by coagulase-negative staphylococci involved in bovine mastitis

Veterinary Microbiology, 2005

In the present study, 188 coagulase-negative Staphylococcus (CNS) strains were isolated from bovine mastitis cases from 56 different Brazilian dairy herds, located in the Southeast region of the country, and were tested for antimicrobial substance production. Twelve CNS strains (6.4%) exhibited antagonistic activity against a Corynebacterium fimi indicator strain. Most antimicrobial substances were sensitive to proteolytic enzymes suggesting that they might be bacteriocins (Bac). Amongst the CNS producers, six were identified as S. epidermidis, two as S. simulans, two as S. saprophyticus, one as S. hominis and one as S. arlettae. Plasmid profile analysis of these strains revealed the presence of at least one plasmid. The Bac + strains presented either no or few antibiotic resistance phenotypes. Three strains were shown to produce a bacteriocin either identical or similar to aureocin A70, a bacteriocin previously isolated from an S. aureus strain isolated from food. The remaining Bac + strains produce antimicrobial peptides that seem to be distinct from the best characterised staphylococcal bacteriocins described so far. Some of them were able to inhibit Listeria monocytogenes, an important food-borne pathogen, and several strains of Streptococcus agalactiae associated with bovine mastitis, suggesting a potential use of these bacteriocins either in the prevention or in the treatment of streptococcal mastitis. #

Phenotypic and genetic characterisation of bacteriocin-producing strains of Staphylococcus aureus involved in bovine mastitis

Veterinary Microbiology, 2002

Fifty strains of Staphylococcus spp. isolated from bovine mastitis cases in several herds from different Argentinian provinces were screened for antimicrobial substances. Twelve strains exhibited a high antagonistic activity against the indicator strain (Corynebacterium ®mi) and were chosen for further characterisation. The antimicrobial substances were sensitive to proteolytic enzymes suggesting that they might be bacteriocins (Bac). These strains were identi®ed as S. aureus by the ampli®cation of the femA gene. Plasmid pro®le analysis of these strains revealed the presence of at least one plasmid. Eleven strains carried a plasmid with a size similar to that of pRJ6 (8.0 kb), which encodes aureocin A70, a bacteriocin produced by the Brazilian S. aureus strain A70 isolated from commercial milk. The other strain harboured a much larger plasmid. PCR experiments, using speci®c primers for ampli®cation of the bacteriocin operon found in pRJ6, showed that all strains had the expected 525 bp amplicon, suggesting that the bacteriocin produced may be related to aureocin A70. The genomic DNA of all Bac strains was then analysed by pulsed-®eld gel electrophoresis (PFGE) in order to investigate clonal relationships amongst strains. Based on the results of PFGE experiments, 10 out of the 12 Bac strains belonged to the same clone. The remaining two strains are possibly related to the prevalent clone. The aureocin A70 producer-strain belonged to a distinct clone. #

Bacteriocins of non-aureus staphylococci isolated from bovine milk

Applied and environmental microbiology, 2017

Non-aureus staphylococci (NAS), the bacteria most commonly isolated from the bovine udder, potentially protect the udder against infection by major mastitis pathogens due to bacteriocin production. In this study, we determined the inhibitory capability of 441 bovine NAS isolates (comprising 26 species) against bovine S. aureus Furthermore, inhibiting isolates were tested against a human methicillin-resistant S. aureus (MRSA) isolate using a cross-streaking method. We determined the presence of bacteriocin clusters in NAS whole genomes using genome mining tools, BLAST, and comparison of genomes of closely related inhibiting and non-inhibiting isolates and determined the genetic organization of any identified bacteriocin biosynthetic gene clusters. Forty isolates from 9 species (S. capitis, S. chromogenes, S. epidermidis, S. pasteuri, S. saprophyticus, S. sciuri, S. simulans, S. warneri, and S. xylosus) inhibited growth of S. aureus in vitro; 23 of which, from S. capitis, S. chromogen...

Enhancing Drug Efficacy against Mastitis Pathogens—An In Vitro Pilot Study in Staphylococcus aureus and Staphylococcus epidermidis

Animals

Background: Bovine mastitis is one of the major infectious diseases in dairy cattle, resulting in large economic loss due to decreased milk production and increased production cost to the dairy industry. Antibiotics are commonly used to prevent/treat bovine mastitis infections. However, increased antibiotic resistance and consumers’ concern regarding antibiotic overuse make it prudent and urgent to develop novel therapeutic protocols for this disease. Materials and methods: Potential druggable targets were found in 20 mastitis-causing pathogens and conserved and unique targets were identified. Bacterial strains Staphylococcus aureus (ATCC 29213, and two clinical isolates CI 1 and CI 2) and Staphylococcus epidermidis (ATCC 12228, and two clinical isolates CI 1 and CI 2) were used in the present study for validation of an effective drug combination. Results: In the current study, we identified the common and the unique druggable targets for twenty mastitis-causing pathogens using an i...

The Efficacy of Bacteriocins Against Biofilm-Producing Bacteria Causing Bovine Clinical Mastitis in Dairy Farms: A New Strategy

Current Microbiology, 2023

Using an alternative bio-product is one of the most promising ways to control bovine mastitis and avoid new intra-mammary infections. The aims of this study were to ascertain the prevalence of biofilm-forming bacteria responsible for causing clinical mastitis in dairy herds and to assess the effectiveness of bacteriocins, produced by Bacillus subtilis, in controlling the growth of these bacteria in the milk of animals. A total of 150 milk samples were collected from cows and buffalos suffering from mastitis and the etiological agents were isolated and identified by the VITEK-2-COMPACT-SYSTEM®. Additionally, the capability of the bacterial isolates to produce biofilms was determined. RT-PCR was used to detect enterotoxin-producing genes (sed and seb), resistance genes (mecA and blaZ), and biofilm-associated genes (icaA and fnbA) in the isolated bacteria. The susceptibility patterns of the bacterial isolates to bacteriocins were assessed using an agar well-diffusion assay. S. aureus was significantly more capable of producing biofilms than coagulase-negative Staphylococcus isolates. S. ubris was the strongest biofilm producer among the Streptococcus species. The sensitivity profiles of the Staphylococcus spp. (S. aureus and coagulase-negative Staphylococcus) and their biofilm producers to bacteriocins were significantly higher (100% and 90%, respectively) at the same concentration. Bacteriocins had a lethal effect on Staphylococci, Streptococci, and biofilm development at a dose of 250 µg/mL. In dairy farms, bacteriocins are a viable alternative treatment for the prevention and control of bovine clinical mastitis. Abbreviation CNS Coagulase-negative Staphylococcus E. faecalis Enterococcus faecalis fnbA Fibronectin-binding protein A gene FESEM Field emission scanning electron microscopy icaA Intercellular adhesion A gene S. aureus Staphylococcus aureus S. agalactiae Streptococcus agalactiae S. dysgalactiae Streptococcus dysgalactiae S. lactarius Streptococcus lactarius S. ubris Streptococcus ubris

Bacteriocin Producing Streptococcus agalactiae Strains Isolated from Bovine Mastitis in Brazil

Microorganisms, 2022

Antibiotic resistance is one of the biggest health challenges of our time. We are now facing a post-antibiotic era in which microbial infections, currently treatable, could become fatal. In this scenario, antimicrobial peptides such as bacteriocins represent an alternative solution to traditional antibiotics because they are produced by many organisms and can inhibit bacteria, fungi, and/or viruses. Herein, we assessed the antimicrobial activity and biotechnological potential of 54 Streptococcus agalactiae strains isolated from bovine mastitis. Deferred plate antagonism assays revealed an inhibition spectrum focused on species of the genus Streptococcus—namely, S. pyogenes, S. agalactiae, S. porcinus, and S. uberis. Three genomes were successfully sequenced, allowing for their taxonomic confirmation via a multilocus sequence analysis (MLSA). Virulence potential and antibiotic resistance assessments showed that strain LGMAI_St_08 is slightly more pathogenic than the others. Moreover,...

Novel Quantitative Assay to Describe In Vitro Bovine Mastitis Bacterial Pathogen Inhibition by Non-aureus Staphylococci

Pathogens, 2022

In this paper, we describe a new quantitative method to evaluate and quantify in vitro growth inhibition of mastitis-related bacteria. Colony-forming units of Staphylococcus (S.) aureus (n = 10), Escherichia (E.) coli (n = 10), and Streptococcus (S.) uberis (n = 10) were quantified after their growth on top of layers of trypticase soy agar (TSA) containing six different concentrations (varying from 102 to 107 CFU/mL) of bovine non-aureus staphylococci (NAS), i.e., S. chromogenes (n = 3) and S. simulans (n = 3) isolates. Growth inhibition of the mastitis-related major bacterial pathogens, including E. coli, was confirmed by all NAS, an effect that varied highly among NAS isolates and was not evident from the semiquantitative method with which the new method was compared. By subsequent application of the new method on a larger set of 14 bovine NAS isolates, we observed that S. simulans and NAS originating from teat apices (especially S. epidermidis) required lower concentrations to in...