Veterinary Research Forum

Abstract This study aimed to obtain a narrow-spectrum antimicrobial peptide. A peptide XMK-8 was designed based on the amino acid sequence of goose MyHC1 protein from positions 1919 to 1936 (some parameters do not meet the requirements of antimicrobial peptides through bioinformatics analysis) using bioinformatics tools and amino acid substitution method. The minimum inhibitory concentration was determined using liquid double dilution method, the hemolysis rate was determined using dilution method, and the effects of temperature, acid-base, enzyme, and salt ions on its antimicrobial activity were evaluated using liquid double dilution method. The results showed that the designed peptide was a cationic hydrophilic peptide with high amphiphilicity and low hemolytic activity on mouse red blood cells. It had no antimicrobial activity against Escherichia coli, Salmonella, Staphylococcus aureus, and Aeromonas hydrophila. The minimum inhibitory concentration against Pasteurella multocida was 250 μg mL^-1, and the minimum inhibitory concentration against Haemophilus parasuis was 1.00 mg mL^-1. The antimicrobial activity of the narrow-spectrum antimicrobial peptide XMK-8 can still be detected after treatment with temperature (0.00 - 100 ˚C), salt ions (sodium ions and potassium ions; 50.00 - 200 mmol L^-1), pH (4.00 - 10.00), and protease K (20.00 - 100 μg mL^-1). Antimicrobial peptide XMK-8 was expected to become a new alternative to antibiotics and would have good application prospects in the prevention and treatment of P. multocida and H. parasuis infections.

Abstract The efficacy of probiotics as alternatives to antibiotics has been defined as one of the potential strategies to prevent Salmonella spp. infection in poultry. The purpose of this study was to isolate probiotic native Lactic acid bacteria (LAB) with high compatibility to intestinal tract and prevention of Salmonella typhimuriumfrom broiler chicken feces. Thirty-seven samples of chicken feces were collected from seven broiler chicken farms in Northern Iran. The isolates identification was carried out with morphological and biochemical tests. Agar diffusion methods were used to evaluate the antimicrobial activities against Escherichia coli and S. typhimurium. The primary probiotic characteristics such as resistance to acid and bile and adhesion to Caco-2 cells were studied. Indeed, the ability of LAB isolates to inhibit adhesion of S. typhimurium to Caco-2 cells was evaluated by exclusion, competition and displacement assays. Among 42 isolates, S08, S01 and S06 isolates which showed appropriate probiotics characteristics were selected. Isolates S08 and S01 showed to be able to adhere strongly and also S06 was adhered moderately. In the exclusion assay, the isolates S08, S01 and S06 significantly hampered adhesion of S. typhimurium cell, in the competition assay, the isolates S08, S01 showed significant level of competition activity against S. typhimurium adherence to Caco-2 cells and isolate S08 showed the greatest displacement activity. The 16S rDNA sequence revealed that S08, S01, and S06 isolates were 99.00% similar to Lactobacillus salivarius, Lactobacillus johnsonii, and Pediococcus acidilactici, respectively. The result of this study suggested that LAB isolated from broiler chicken feces could be a remarkable reservoir for identification of probiotic to inhibit the pathogenic bacteria growth.

Abstract In the present study, the effect of different concentrations of cell-free supernatant (CFS; 10.00 and 35.00 mg g^-1) of Lactobacillus salivarius (Ls-BU2) on chemical, microbial and sensorial specifications of ground beef stored under the refrigerated condition was investigated. The antibacterial activity of CFS on Escherichia coli was also assessed. According to agar-disk diffusion method, CFS of Ls-BU2 revealed a promising antibacterial activity against E. coli in culture media compared to CFS of a well-known probiotic (L. acidophilus LA-5). In meat, CFS of Ls-BU2 showed a minimal effective concentration (MEC) of 35.00 mg g^-1 on E. coli, while CFS of L. acidophilus represented a MEC of > 45.00 mg g^-1. The CFS of Ls-BU2 at 35.00 mg g^-1 concentration retained psychrophilic counts of meat at a lower value than maximum accepted level (7 log₁₀ CFU g^-1). In a similar trend, CFS of Ls-BU2 at 35.00 mg g^-1 concentration was also displayed high sensorial scores compared to other CFS-treated samples. In conclusion, we demonstrated that CFS of Ls-BU2 and to some extent CFS of L. acidophilus could act as a safe food additive for the control of bacterial pathogens and to extend the shelf life of ground beef.

Abstract Echinophora Platyloba D.C as a medicinal plant is used for preservation of foods and treatment of many diseases in different regions of Iran. The present study was undertaken to determine the chemical composition and investigation of the antibacterial effects of essential oil as well as methanol extract from aerial part of Echinophora Platyloba D.C against S. aureus, L. monocytogenes, S. Thyphimurium and E. coli. Chemical analysis using gas chromatography and mass spectrophotometry (GC/MS) showed that ocimene (26.51%), 2,3-Dimethyl-cyclohexa-1,3-diene (9.87%), alpha-pinene (7.69%) and gamma-dodecanolactone (5.66%) were dominant components of essential oil and the main constituents of methanol extract were o-Cymene (28.66%), methanol (8.50%), alpha-pinene (7.42%) and gamma-decalactone (5.20%). The essential oil showed strong antimicrobial activity against tested bacteria, whereas the methanol extract almost remained inactive against gram-negative bacteria. The most sensitive bacteria to essential oil and extract of Echinophora Platyloba D.C were L. monocytogenes and S. aureus. Minimum inhibitory concentration (MIC) values of essential oil against L. monocytogenes and S. aureus were 6250 and 12500 ppm, respectively. MIC of methanol extract against S. aureus and L. monocytogenes was 25000 ppm. Therefore, purifying and evaluation of antibacterial effects of the active substances of the essential oil and methanol extract of this plant for future application as antibacterial agents and food preservatives to combat pathogenic and toxigenic microorganisms is recommended.