Changes in bovine milk bacterial microbiome from healthy and subclinical mastitis affected animals of the Girolando, Gyr, Guzera, and Holstein breeds.

Raw milk samples were collected from 200 dairy cows belonging to Girolando 1/2, Gyr, Guzera, and Holstein breeds, and the bacterial diversity was explored using 16S rRNA amplicon sequencing. SCC analysis showed that 69 animals were classified as affected with subclinical mastitis. The milk bacterial microbiome was dominated by Firmicutes, Proteobacteria, and Actinobacteria, with an increase of Firmicutes in animals with subclinical mastitis and Proteobacteria in healthy animals. At the family and genus level, the milk bacterial microbiome was dominated by Staphylococcus, Acinetobacter, Pseudomonas, members of the family Enterobacteriaceae, Lactococcus, Aerococcus, members of the family Rhizobiaceae, Anaerobacillus, Streptococcus, members of the family Intrasporangiaceae, members of the family Planococcaceae, Corynebacterium, Nocardioides, and Chryseobacterium. Significant differences in alpha and beta diversity analysis suggest an effect of udder health status and breed on the composition of raw bovine milk microbiota. LEfSe analysis showed 45 and 51 discriminative taxonomic biomarkers associated with udder health status and with one of the four breeds respectively, suggesting an effect of subclinical mastitis and breed on the microbiota of milk in cattle.

Prospecting of potentially probiotic lactic acid bacteria from bovine mammary ecosystem: imminent partners from bacteriotherapy against bovine mastitis.

Mastitis is one of the most important causes of loss of cattle production, burdening producers due to the increased cost of milk production and decreased herd productivity. The development of alternative methods for the treatment and prevention of mastitis other than traditional chemical antibiotic therapy needs to be implemented to meet international pressures to reduce the use of these drugs and promote the elimination of multiresistant microbial strains from the environment. Treatment with probiotic bacteria or yeast strains offers a possible strategy for the control of mastitis. The objective of this work was to isolate, identify, and characterize lactic bacteria from milk and the intramammary duct of Gyr, Guzerat, Girolando 1/2, and Holstein cattle breeds from Brazil. Samples of 115 cows were taken, a total of 192 bacteria isolates belonging to 30 species were obtained, and 81 were selected to evaluate their probiotic potential in in vitro characterization tests. In general, bacteria isolated from the mammary gland have low autoaggregation, cell surface hydrophobicity, and co-aggregation with mastitis etiological bacteria Staphylococcus aureus and Escherichia coli. Also, they have biofilm assembly capacity, inability to produce exopolysaccharides, high production of H2O2, and strong antagonism against mastitis pathogens. Ten lactic bacteria isolates were used in co-culture with human MDA-MB-231 breast epithelial cells to assess their adhesion capacity and impairment of the S. aureus invasion. Our results, therefore, contribute to the future production of new prevention and treatment tools for bovine mastitis.

Plants of the Cerrado with antimicrobial effects against Staphylococcus spp. and Escherichia coli from cattle.

BACKGROUND: Both diarrhea in calves and mastitis in cows limit cattle production. The bacteria involved in these diseases have shown multi-resistance to antimicrobials, however plant metabolites therefore can provide an alternative method of control. This study selected and characterized Cerrado plant extracts showing inhibitory effects against Escherichia coli and Staphylococcus spp. from cattle. Thirteen leaf extracts were initially screened and diameters of inhibition zones produced against the pathogens were recorded using an agar disk diffusion method. Total condensed tannin contents were determined and antibacterial activities were analyzed after tannin removal from the five selected extracts. The minimum inhibitory concentrations (MIC) and minimum bactericidal concentrations (MBC) were evaluated by macro-dilution antimicrobial susceptibility tests, and the extracts were characterized by high performance liquid chromatography. RESULTS: Inter- and intra-specific bacterial variations in the susceptibility to the extracts were detected. The aqueous extract (AE) from Caryocar brasiliense Cambess. leaves produced larger inhibition zones against E. coli strains than did other selected extracts. However, the AE from Schinopsis brasiliensis was the most effective against Staphylococcus spp. strains (P < 0.001). The MIC of ethanolic extracts (EE) from C. brasiliense (0.27 mg/mL) and S. brasiliensis (0.17 mg/mL) were lower than those of other extracts. The MIC and MBC of the Annona crassiflora EE were 6.24 mg/mL for all bacteria. Flavonoids were the main metabolites detected in the A. crassiflora EE as well as in the AE and EE from C. brasiliense, while tannins were the main metabolites in the S. brasiliensis leaf extracts. CONCLUSION: The AE from C. brasiliense was more effective against Gram-negative bacteria, while the AE from S. brasiliensis was more effective against Gram-positive bacteria. A. crassiflora EE and S. brasiliensis extracts are potent bactericide. After removal of the tannins, no antimicrobial effects were observed, indicating that these metabolites are the main active antibacterial components.

Short communication: Survival of Vaccinia virus in inoculated cheeses during 60-day ripening.

Bovine vaccinia is a neglected zoonosis caused by Vaccinia virus (VACV) and has a major economic and public health effect in Brazil. Previous studies showed infectious VACV particles in milk from either experimentally or naturally infected cows and in fresh cheeses prepared with experimentally contaminated milk. Ripening is a process that leads to major changes in the physical and chemical characteristics of cheese, reducing contamination by spoilage, pathogenic microorganisms, or both. However, it is not known if VACV infectious particles persist after the ripening process. To investigate this issue, viral infectivity at different ripening times was studied in cheeses manufactured with milk experimentally contaminated with VACV strain Guarani P2 (GP2). Cheeses were analyzed at 1, 7, 14, 21, 45, and 60 d of ripening at 25°C. Viral DNA was quantified by real-time PCR, and VACV isolation and titration were performed in Vero cells. The whole experiment was repeated 4 times. Analysis of the mean viral DNA quantification and infectivity indicated a reduction of approximately 2 logs along the ripening process; however, infectious viral particles (1.7 × 102 pfu/mL) could still be recovered at d 60 of ripening. These findings indicate that the ripening process reduces VACV infectivity, but it was not able to inactivate completely the viral particles after 60 d.

Effects of undernutrition on glycine metabolism in the cerebellum of rats.

Undernutrition is a worldwide problem affecting millions of unborn and young children during the most vulnerable stages of brain development. Total restriction of protein during the perinatal period of life can alter the development of the mammalian fetus and have marked repercussions on development of the central nervous system (CNS). The brain is vulnerable to undernutrition with altered morphologic and biochemical maturation, leading to impaired functions. The focus of this study is to investigate [U-14C]glycine metabolism in undernourished rats submitted to pre- and postnatal protein deprivation (diet: 8% protein with and without addition of L-methionine; control group: 25% protein). Although undernutrition produced a reduction in cerebellar weight and alterations in the DNA concentration, the present study shows that glycine metabolism in this structure is partially protected because the undernourished group with L-methionine did not show modifications in glycine metabolism at all ages studied. However, L-methionine deficiency alters glycine metabolism at 7 and 21 days, but in the adult age both undernourished groups presented no differences in oxidation to CO2, conversion to lipids and incorporation into protein from glycine, compared to the control group.