The Staphylococcus aureus is the most common isolated microorganism in ruminant animal species diagnostic with clinical or subclinical mastitis. Dairy herds with these diseases can transfer S. aureus into the milk supply, which can lead to food poisoning in humans. The objective of this study was to evaluate the profile of antimicrobial susceptibility, the presence of femA gene, the genetic relationships among isolates of S. aureus obtained from milk originating from flocks diagnosed with subclinical mastitis in nine rural properties in the northern of Minas Gerais State. To this end, 498 samples of bovine milk tested positive for the California mastitis test (CMT) were subjected to morphological methods and biochemical patterns for microbiological presumptive identification of S. aureus. The PCR test with the genetic marker femA was used to confirm the species S. aureus. All the 26 isolates presumptively identified as S. aureus amplified a fragment of 132 bp corresponding to the femA gene. The profile of antimicrobial susceptibility was performed according to the disk-diffusion methodology and two isolates were susceptible to all the antibiotics tested. The drug multiresistence was found in 80.76% of the isolates. The determination of the genetic profile and the clonal relationship among the isolates was performed by the method of DNA RAPD-PCR polymorphism. The S. aureus isolates were divided into two groups with 26 distinct subgroups. The analysis of RAPD-PCR showed no genetic diversity among them, heterogeneous profile and absence of clonality.
Studies about preservative and antioxidant activity of essential oils have been encouraged in recent years, given their importance to food industry. The aim of the current study was to evaluate the antioxidant properties and antimicrobial activity of essential oils deriving from Syzygium aromaticum, Cymbopogon citratus and Lippia alba against lactic and pathogenic bacteria responsible for food-borne diseases. Essential oil antibacterial activity was assessed through disc diffusion and macrodilution tests conducted in a mixed lactic culture of Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus (YF-L903) and of Escherichia coli (ATCC 8739), Staphylococcus aureus (ATCC 6538), Salmonella enterica (ATCC 6017) strains. Based on the chromatographic analysis results, the essential oils shown to be composed of eugenol (79.41%) which was the prevalent compound in S. aromaticum, geranial (31.89%), neral (24.52%) and β-myrcene (25.37%) in C. citratus, as well as of geranial (33.80%) and neral (25.63%) in L. alba. The observed antibacterial activity confirmed the dose-dependent effect of these three oils on all the assessed bacteria; there was halo inhibition at concentration 20μL mL-1. The essential oil of S. aromaticum presented better antioxidant activity, with IC50 equal to 5.76μg mL-1 and antioxidant activity index of 6.94, and it was considered strong (AAI>2.0) in comparison to the other evaluated oils. This essential oil also presented excellent antioxidant activity at concentrations lower than the one required to inhibit lactic cultures. Based in this outcome, the essential oil from S. aromaticum can be used as preservative agent in processed food whose formulation presents lactic cultures.
O uso de antimicrobianos como melhoradores de desempenho tem representado um risco à saúde pública devido à associação destes com o surgimento de resistência bacteriana e resíduos nos produtos. Como alternativa, pesquisas têm indicado a utilização de aditivos alimentares naturais, como o óleo essencial de capimlimão. Objetivou-se neste artigo avaliar a atividade antimicrobiana do óleo essencial de capim-limão (Cymbopogon flexuosus) frente a Staphylococcus aureus e Escherichia coli entéricas isoladas de aves de postura (Gallus gallus domesticus). Foram preparadas cinco concentrações do óleo essencial (160; 80; 40; 20 e 10 µL mL-1) e então realizados testes in vitro de difusão em disco e determinação da Concentração Inibitória Mínima (CIM) e da Concentração Bactericida Mínima (CBM). Os resultados indicaram que a concentração de 160 µL mL-1 propiciou os maiores diâmetros de halos de inibição na difusão em disco, diferindo-se estatisticamente das demais concentrações (p < 0,05). Houve efeito inibitório (CIM) a partir da concentração de 80 µL mL-1 de óleo para todas as bactérias mas a CBM não foi detectada. Conclui-se que o óleo essencial de capim-limão apresenta atividade antimicrobiana in vitro frente a S. aureus e E. coli isoladas de aves na concentração de 160 µL mL-1. Novos estudos são necessários para avaliar a eficácia do uso de doses superiores e indicar esse óleo como aditivo em dietas de aves.
SUMMARY The antibacterial effect of microencapsulated lemon grass (Cymbopogon citratus) essential oil on strains of Escherichia coli (ATCC8739), Staphylococcus aureus (ATCC 6538) and Salmonella enterica subsp. enterica (ATCC 6017), and the stability of this oil in feeds for broiler chickens were evaluated. The Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) were determined by the macrodilution method, using the microencapsulated lemon grass essential oil at concentrations of 160 μL mL−1, 80 μL mL−1, 40 μL mL−1, 20 μL mL−1, and 10 μL mL−1. The oil concentration of 80 μL mL−1 presented the best results against the three bacteria evaluated. Samples of 200 g of feed mixed with 120 μL g−1 of the microencapsulated lemon grass essential oil was stored to evaluate the oil stability. Feed without microencapsulated lemon grass essential oil was prepared as control. The oil remained active for seven days, with significant reduction of S. aureus (3.08 CFU), E. coli (3.01 CFU), and S. enterica (3.10 CFU). The microencapsulated lemon grass essential oil at concentration of 80 μL mL−1 had antibacterial effect against the E. coli, S. enterica and S. aureus, and maintained stability of the feed for seven days, even with presence of organic matter, which is source of nutrients for pathogens.
Background and Aim: The use of antimicrobials in the control of mastitis is of concern in public health due to their inefficiency in targeting microorganisms. Studies with medicinal plants have risen as an alternative to the use of conventional products. The objective of this study was to evaluate the efficacy of an experimental disinfectant based on the essential oil (EO) from Lippia origanoides in preventing the development of new intramammary infections (IMI) in Holstein cows. Materials and Methods: The conventional protocol of pre- and post-milking was used and the control (Conventional treatment [CNV]) and experimental (Experimental treatment [PEX]) products containing EO at 120 μL/mL were applied by immersion. Individual milk samples were analyzed using sheep blood agar methodologies and biochemical tests. The efficiency of the treatment was defined by the presence or absence of Staphylococcus aureus, coagulase-negative Staphylococcus, and Streptococcus spp. Results: There were no clinical and subclinical mastitis cases, no lesions in the mucosal of teats, nor dirt score between groups in this study. Both treatments did not influence the occurrence of IMI. Conclusion: The results revealed that PEX acts efficiently against microorganisms compared to the disinfection by the conventional product demonstrating the efficacy of the alternative product on the prevention of new IMIs in dairy cows.
905RESUMO.-Objetivou-se determinar atividade antisséptica do óleo essencial de Lippia origanoides na presença de leite bovino. A composição química do óleo essencial de alecrim pimenta foi determinada por cromatografia gasosa acoplada à espectrometria de massas (CG-EM), sendo detectados 53 compostos, dos quais 16 foram identificados (>0,1% área total). O carvacrol (32,7%), p-cimeno (23%), timilmetil éter, cariofileno (7,98%) e o γ-terpineno (5,40%) foram os componentes mais abundantes. A concentração inibitória mínima para Staphylococcus aureus ATCC 25923 e E. coli ATCC 8739 foi de 60µL/mL, enquanto para Salmonella Choleraesuis ATCC 35640 foi de 90µL/mL. A concentração bactericida mínima foi de 120µL/mL para as três bacté-rias. Avaliou-se a atividade antisséptica do óleo essencial na concentração de 120µL/mL na presença de leite bovino sobre as mesmas cepas bacterianas. O óleo em estudo apresentou efeito inibitório do crescimento das cepas em diferentes tempos de ação (p<0,05). S. aureus apresentou maiores índices de inibição após 5 min de contato e Escherichia coli e Salmonella Choleraesuis após 15 min. O óleo essencial de alecrim-pimenta revelou ser um potencial antimicrobiano natural, mesmo na presença de matéria orgâ-nica constituída de uma matriz nutricional complexa que é o leite bovino. This study aimed to determine the antiseptic activity of the essential oil of Lippia origanoides in the presence of bovine milk. The essential oil chemical composition was determined by gas chromatography-mass spectrometry (CG EM) 53 compounds were detected, of which 16 were identified (>0.1% total area). The carvacrol (32.7%), p-cymene (23%), thymil methyl ether (10.03%), caryophyllene (7.98 %) and γ-terpinene (5.40%) were the most abundant components. The minimum inhibitory concentration for Staphylococcus aureus ATCC 25923 and Escherichia coli ATCC 8739 was 60uL/mL, as for this Salmonella Choleraesuis ATCC 35640 was 90uL/mL. The minimum bactericidal concentration was 120μL/ mL for all three bacteria. We evaluated the antiseptic activity of the essential oil in the concentration of 120μL/mL in the presence of bovine milk for the same bacterial strains. The oil under study It showed an inhibitory effect of growth of the strains in different action times (p <0.05). S. aureus showed higher inhibition rates after 5 min of contact and E. coli and Salmonella Choleraesuis after 15 min. The essential oil Lippia origanoides proved to be a natural antimicrobial potential even in the presence of organic matter consists of a complex nutritional matrix is bovine milk.
SUMMARY The effect of the disinfection of fertile eggs of free-range poultry with essential oils from Cymbopogon flexuosus and Lippia rotundifolia was evaluated. The treatments applied to the eggs were (v v-1): 1% essential oil from C. flexuosus (CF), 1% essential oil from L. rotundifolia(LR), 0.1% CF + LR, and 0.1% quaternary ammonia (QA) (positive control). The eggs were incubated for 21 days, the birds from hatched eggs were counted and weighed, and not hatched eggs were opened and examined macroscopically to determine egg fertility, and embryonic death time. The antimicrobial action of the essential oils was determined through quantitative analyses, considering aerobic mesophylls, filamentous fungi, yeasts, and total coliforms. Mortality was high in the treatments, especially in the treatment with essential oil from C. flexuosus. The highest hatchability was found in the treatment with QA. The treatments with singly or combined essential oils were more efficient in decreasing microbial count than QA. The essential oils from C. flexuosus and L. rotundifolia reduced the microbial load of fertile eggs of the free-range poultry evaluated.
Background and Aim: Broiler meat is susceptible to lipid oxidation due to its high content of unsaturated fatty acids, especially when stored for a long time. Concerning with that problem, we can add natural compounds to the animal feed, as the plant essential oils, which have antioxidant, antimicrobial, and antifungal activities. The objective of this study was to evaluate the antioxidant activity, fatty acid profile, and chemical composition of the meat of broilers fed with diets containing different essential oil, including lemongrass (Cymbopogon flexuosus) and pedestrian tea (Lippia aff. rotundifolia). Materials and Methods: The experimental design used was a 4×4 factorial scheme (storage time×diets), and each treatment was repeated thrice. The treatments were applied as negative control (without antimicrobial); positive control (ration supplemented with 10 ppm of enramycin and 42 ppm of salinomycin), lemongrass (ration with 120 mg of essential oil of lemongrass for each kilogram of live weight of the animals), and pedestrian tea (ration with 120 mg of essential oil of pedestrian tea for each kilogram of live weight of the animals). Storage was performed after slaughter and evaluated monthly for 4 months. Results: The inclusion of lemongrass oil and pedestrian tea in the broiler diet did not change the ether extract content of the meat sample obtained from thigh and drumstick. The storage time promotes an increase in the moisture loss of the meat from the 2nd month in all treatments. There was an interaction between the additive and storage time for moisture loss. In the 2nd month of storage, the meat from broilers fed the antibiotic-containing feed had a lower moisture loss than those from broilers in the other treatment groups. The fatty acid profile did not differ statistically between the different treatment groups. The inclusion of additives in the food dropped the lipid oxidation at the 2nd month of storage. From the 3rd month, however, only the essential oils showed antioxidant effect. Conclusion: Antioxidant activity was positive for treatments that included essential oils. The lemongrass oil and pedestrian tea can be used as antioxidant additives in broiler diets to improve the oxidative stability of thigh-drumstick meat during storage. The results of the study suggest a good prospective for further research with profitability of usage essentials oils examined, for their economical application as feed components in broiler nutrition.
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