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. 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. 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.
Fungi have the ability to degrade vegetal cell wall carbohydrates, and their presence in the digestive tract of ruminants can minimize the effects of lignified forage on ruminal fermentation. Here, we evaluated enzyme production by Aspergillus spp. isolates from the digestive tracts of cattle grazed in tropical pastures during the dry season. Filamentous fungi were isolated from rumen and feces by culture in cellulose-based medium. Ninety fungal strains were isolated and identified by rDNA sequence analysis, microculture, or both. Aspergillus terreus was the most frequently isolated species, followed by Aspergillus fumigatus. The isolates were characterized with respect to their cellulolytic, xylanolytic, and lignolytic activity through qualitative evaluation in culture medium containing a specific corresponding carbon source. Carboxymethyl cellulase (CMCase) activity was quantified by the reducing sugar method. In the avicel and xilan degradation test, the enzyme activity (EA) at 48 h was significantly higher other periods (P < 0.05). Intra- and inter-specific differences in EA were verified, and high levels of phenoloxidases, which are crucial for lignin degradation, were observed in 28.9% of the isolates. Aspergillus terreus showed significantly higher EA for avicelase (3.96 ±1.77) and xylanase (3.13 ±.091) than the other Aspergillus species at 48 h of incubation. Isolates AT13 and AF69 showed the highest CMCase specific activity (54.84 and 33.03 U mg-1 protein, respectively). Selected Aspergillus spp. isolates produced remarkable levels of enzymes involved in vegetal cell wall degradation, suggesting their potential as antimicrobial additives or probiotics in ruminant diets.
Human ingestion of antimicrobial residues in dairy products is a problem of public health. In this study we evaluated antibacterial effectiveness of aqueous and ethanolic leaf extracts of Mangifera indica L. as a natural alternative for reduction of Staphylococcus spp. biofilms. Eight Staphylococcus spp. strains from cows with mastitis and the Staphylococcus aureus strain (ATCC 25923) were evaluated. High performance liquid chromatography indicated the presence of phenolic compounds in the aqueous extract. Agilent Technologies gas chromatography revealed the presence of phenolic compounds, carbohydrates, and gallic acid in ethanolic extract. The tannin contents were 2.27 ± 0.01 and 2.67 ± 0.01% for ethanolic extract and aqueous extract, respectively. Extracts were evaluated in agar diffusion test and the size of inhibition zones were compared. The ethanolic extract was the most effective, showing minimum inhibitory concentrations and minimum bactericidal concentrations between 1.8–7.5 and 15.1–45.3 mg/ml, respectively. After tannin removal, the extracts were less effective, indicating these secondary metabolites as one of the main antibacterial compounds. The ethanolic extract at 45.3 mg/ml reduced mature biofilms in teat rubber (average: 3.91 log reduction) and stainless steel (average: 3.87 log reduction) after 5 min of contact. The extracts of M. indica leaves represent natural alternatives against Staphylococcus spp. strains and the ethanolic extract shows potential as a natural sanitizer.