Mode of action and in vitro susceptibility of mastitis pathogens to macedocin ST91KM and preparation of a teat seal containing the bacteriocin

Mastitis is considered to be the most economically costly disease affecting the dairy industry. Regular dosage of animals with antibiotics, including use of prophylactic concentrations, may select for resistant strains. The purpose of this study was to determine the mode of action of a new bacteriocin (macedocin ST91KM), to evaluate the antimicrobial resistance of mastitis pathogens to antibiotics commonly used in treatment remedies, and to introduce the possible use of an alternative antimicrobial agent. The bacteriocin macedocin ST91KM, produced by Streptococcus gallolyticus subsp. macedonicus ST91KM, is bactericidal to Streptococcus agalactiae, Streptococcus dysgalactiae, Streptococcus uberis and Staphylococcus aureus associated with mastitis infections, including strains resistant to methicillin and oxacillin. Sensitive cells were deformed and secreted nucleotides, K+ and â-galactosidase when exposed to macedocin ST91KM. Adsorption of the peptide to target cells decreased in the presence of solvents, suggesting that receptors on the cell surfaces have lipid moieties. No adsorption was recorded in the presence of MgCl2, KI and Na2CO3, suggesting that ionic strength plays an important role. A teat seal preparation containing macedocin ST91KM effectively released the peptide and inhibited the growth of S. agalactiae. Macedocin ST91KM could form the basis for alternative dry cow therapy to prevent mastitis infections in dairy cows as it is effective against pathogens that display resistance to conventional antibiotic therapy.

Bacteriocin production by Lactobacillus pentosus ST712BZ isolated from boza

Bacteriocin ST712BZ (14.0kDa in size) inhibits the growth of Lactobacillus casei,Escherichia coli, Pseudomonas aeruginosa,Enterococcus faecalis, Klebsiella pneumoniae and Lactobacillus curvatus. Growth of strain ST712BZ in BHI, M17, soy milk and molasses was similar to growth in MRS, with optimal bacteriocin production (12800AU/mL) recorded in MRS after 24h. The same level of bacteriocin production (12800AU/mL) was recorded in MRS broth with an initial pH of 6.5, 6.0 and 5.5. However, MRS broth (pH 6.5) supplemented with 1mM EDTA, yielded only 6400AU/mL. Low levels of bacteriocin activity were recorded in MRS broth with an initial pH of 5.0 and 4.5. Of all media compositions tested, MRS supplemented with tryptone (20.0g/L), glucose (20.0 to 40.0g/L), mannose (20.0g/L), vitamin B12, or vitamin C yielded 12800AU/mL. Glycerol concentrations of 1.0g/L and higher repressed bacteriocin production. Maximal bacteriocin activity (25600AU/mL) was recorded in MRS supplemented with Vit. B1 or DL-6,8-thioctic acid.

A bacteriocina ST712BZ produzida par Lactobacillus pentosus (peso molecular de 14,0kDa) inibe o crescimento de Lactobacillus casei,Escherichia coli, Pseudomonas aeruginosa,Enterococcus faecalis, Klebsiella pneumoniae e Lactobacillus curvatus. O crescimento de L. pentosus ST712BZ em BHI, M17, leite de soja e melaços foi semelhante ao observado em MRS, registando-se a produção máxima de bacteriocina (12800UA/mL) em MRS após 24 h. Observou-se o mesmo nível de produção de bacteriocina (12800UA/mL) em caldo MRS com pH inicial de 6,5, 6,0 e 5,5. No entanto, em caldo MRS (pH 6,5) suplementado com 1 mM de EDTA a produção apenas atingiu 6400UA/mL. Os níveis de atividade bacteriocinogênica detectados em caldo MRS com um pH inicial de 5,0 e 4,5 foram baixos. De todas as fórmulas de meios de cultura testadas a que apresentou a atividade máxima 12800UA/mL foi MRS suplemento de triptona (20,0g/L), glicose (20,0 e 40,0 g/L), manose (20.0 g/L), vitamina B12 e vitamina C. A produção de bacteriocinas foi inibida por concentrações de glicerol superiores ou iguais a 1.0 g/L. Verificou-se a actividade bacteriocinogênica máxima (25600UA/mL) em caldo MRS suplementado com vitamina B1 ou ácido DL-6,8-thioctic.