Biocontrol capacity of bacteria isolated from sawdust of the dairy cattle production environment.

This research paper aimed to find endemic bacteria from the cattle production system to control the growth of mastitis pathogens. Bacteria were isolated from compost barn sawdust of two dairy cattle systems and later tested to verify their ability to control the growth of Staphylococcus aureus isolates obtained from cattle with mastitis. Bacterial isolates from these systems were tested to verify biocontrol capacity using the double-layer method. A total of 189 isolates were obtained from all samples by considering the morphology of the different bacterial colonies, with 30 isolates showing positive results for the growth control of at least one S. aureus strain and 19 isolates showing the ability to control more than one pathogen strain. The ability to control more than one pathogen and present a significant halo of inhibition in our isolates represents positive traits in the search for cattle mastitis biocontrol microorganisms. Thus, the results obtained represent the range of bacteria capable of controlling the pathogens without the use of antibiotics.

Effects of the natural antimicrobial peptide aureocin A53 on cells of Staphylococcus aureus and Streptococcus agalactiae involved in bovine mastitis in the excised teat model.

Aureocin A53 is an N-formylated antimicrobial peptide (AMP) produced by Staphylococcus aureus. Aureocin A53 has a broad spectrum of antimicrobial activity against human and animal pathogens. In the present study, its antagonistic activity was investigated towards 30 strains of S. aureus and 30 strains of Streptococcus spp. isolated from bovine mastitis cases in Brazil. Bovine mastitis is a disease that causes a major economic impact worldwide. Aureocin A53 inhibited the growth of all 60 strains tested, including multidrug-resistant streptococcal isolates and strains of S. aureus belonging to different pulsotypes. This AMP proved to be bactericidal against the six target strains randomly selected among staphylococci and streptococci, also exhibiting a lytic mode of action against the staphylococcal cells. Furthermore, it was determined that 2,048 AU/mL of the AMP were required to inhibit 99.99% of the cell growth of the strain less sensitive to aureocin A53. Aureocin A53 was not toxic to bovine mammary gland epithelial cells after a 24-h exposure and maintained its antimicrobial activity when tested in the excised-teat model against strains of S. aureus and Streptococcus agalactiae, the species responsible for most intramammary infections, not only in Brazil but in other countries as well. Therefore, the use of aureocin A53 in the development of new pharmacological products for the prophylaxis and/or treatment of bovine mastitis was considered promising.

A time series analysis of bulk tank somatic cell counts of dairy herds located in Brazil and the United States / Uma análise de séries temporais de contagem de células somáticas de tanque de rebanhos leiteiros localizados no Brasil e nos Estados Unidos

ABSTRACT: Bulk tank somatic cell counts (BTSCC) is widely used to monitore the mammary gland health at the herd and regional level. The BTSCC time series from specific regions or countries can be used to compare the mammary gland health and estimate the trend of subclinical mastitis at the regional level. Three time series of BTSCC from dairy herds located in the USA and the Southeastern Brazil were evaluated from 1995 to 2014. Descriptive statistics and a linear regression model were used to evaluate the data of the BTSCC time series. The mean of annual geometric mean of BTSCC (AGM) and the percentage of dairy herds with a BTSCC greater than 400,000 cells mL-1 (%>400) were significantly different (P<0.05) according to the countries and the times series. Linear regression model used for the USA time series was statistically significant for AGM and the %>400 (P<0.05). The first and second USA time series presented an increasing and decreasing trend for AGM and the %>400, respectively. The linear regression model for the Brazil time series was not significant (P>0.05) for both dependent variables (AGM and %>400). The Brazil time series showed no increasing or decreasing trend for the AGM and %>400. Consequently, approximately 40 to 50% of the dairy herds from southeastern Brazil will not achieve the regulatory limits for BTSCC over the next years.

RESUMO: Contagens de células somáticas de tanque (BTSCC) são amplamente utilizadas para monitorar a saúde da glândula mamária em níveis regional e de rebanho. Séries temporais de BTSCC, de regiões ou países específicos, podem ser usadas para comparar a saúde da glândula mamária e estimar a tendência de mastite subclínica em nível regional. Três séries temporais de BTSCC de rebanhos leiteiros, localizados nos EUA e na região sudeste do Brasil, foram avaliadas de 1995 a 2014. A estatística descritiva e modelos de regressão linear foram usados para avaliar as informações das séries temporais de BTSCC. A média anual geométrica de BTSCC (AGM) e a porcentagem de rebanhos com BTSCC, maior que 400.000 células mL-1 (%>400), foram significantemente diferentes (P<0,05) de acordo com os países e as séries temporais analisadas. O modelo de regressão linear usado para as séries temporais dos EUA foram estatisticamente significantes para AGM e %>400 (P<0,05). A primeira e a segunda séries temporais dos EUA apresentaram uma tendência de aumento e diminuição, tanto para AGM quanto para %>400, respectivamente. O modelo de regressão linear para as séries temporais do Brasil não foi significante (P>0,05) para ambas as variáveis dependentes (AGM e %>400). As séries temporais do Brasil não apresentaram tendência de aumento ou diminuição para AGM e %>400. Consequentemente, aproximadamente, 40 a 50% dos rebanhos leiteiros do sudeste do Brasil não alcançarão os limites regulatórios para BTSCC ao longo dos próximos anos.

Presence of mecA-positive multidrug-resistant Staphylococcus epidermidis in bovine milk samples in Brazil.

Bacteria of the genus Staphylococcus are one of the major pathogens causing bovine mastitis. In recent decades, resistance of this genus to oxacillin (methicillin) has been a matter of concern due to the possibility of reducing the effectiveness of mastitis treatments and the transfer of resistance determinants to other bacteria. Oxacillin resistance was studied in 170 staphylococci from bovine milk samples, including 79 Staphylococcus aureus and 91 coagulase-negative staphylococci (CNS). The susceptibility profile of 10 antimicrobial agents used in veterinary practice was determined by the Etest method. In addition to the Etest, the phenotypic characterization of oxacillin resistance was tested using the cefoxitin disk diffusion test. All isolates were screened by PCR to detect the mecA gene in 2 different regions of the gene. The isolates with an oxacillin minimum inhibitory concentration ≥0.5 µg/mL or resistant to cefoxitin were identified by sequencing a 536-bp fragment of the 16S rRNA gene. This group of isolates was also evaluated for the presence of blaZ and mecC genes. Molecular analysis of the mecA gene was carried out by typing of the staphylococcal cassette chromosome mec (SCCmec). The relatedness of the mecA-positive isolates was evaluated by macrorestriction of chromosomal DNA followed by pulsed-field gel electrophoresis. With the exception of penicillin and oxacillin, 86% of the isolates showed susceptibility to cephalothin, gentamicin, erythromycin, sulfonamide, trimethoprim-sulfamethoxazole, and tetracycline. All S. aureus isolates were susceptible to oxacillin, whereas 47% (n=43) of the CNS isolates were resistant. The CNS isolates showed a higher resistance to cephalothin, erythromycin, tetracycline, and gentamicin in comparison with S. aureus. The mecA gene was only detected in 10 CNS isolates, identified as Staphylococcus epidermidis, and classified into 3 pulsotypes (A, B, and C) and 4 subtypes (A1, B1, B2, and B3). Among the isolates with an oxacillin resistance phenotype, 12 were positive for the blaZ gene, and 9 of them were mecA-positive. Two of the oxacillin-resistant isolates amplified the mecA homolog gene of Staphylococcus sciuri and none amplified mecC. Three SCCmec types, I, IV, and V, were found. Our results suggest that Staphylococcus epidermidis can be a reservoir for mecA for other Staphylococcus species. Studies investigating the molecular and phenotypic profile of antimicrobial resistance in staphylococcal species should be performed for controlling the spread of resistance and the selection of appropriate therapeutic measures.