Tetracycline resistance in lactobacilli isolated from Serbian traditional raw milk cheeses.

The aim of this study was to investigate the presence of tetracycline resistance in lactobacilli isolated from traditional Serbian white brined raw milk cheeses (Homolje, Sjenica, Zlatar). Isolation of presumptive lactobacilli was initially performed using MRS-S agar without tetracycline, or supplemented with 16 and 64 µg/mL of tetracycline. Rep-PCR (GTG)5 genotyping showed a high diversity of the isolates obtained, as examination of 233 isolates resulted in 156 different Rep-PCR fingerprints. Ninety out of 156 (57.69%) of the strains, representatives with different (GTG)5 fingerprints, were identified by MALDI-TOF MS as lactobacilli, while 66 out of 156 (42.31%) strains were identified as members of other LAB genera. All except one out of 90 Lactobacillus isolates further tested by microdilution method, demonstrated unimodal distribution of tetracycline MIC values which were equal to or lower from the breakpoint MIC values (EFSA in EFSA J 10: 1-10, 2012. 10.2903/j.efsa.2012.2740). Only one Lb. paracasei isolate showed the presence of tet(M) gene, while the other analyzed tet genes [tet(A), tet(B), tet(C) tet(K), tet(L), tet(O) and tet(W)] were not detected in any of the isolates. The results of this study indicates that lactobacilli from traditional Serbian raw milk cheeses do not present considerable tetracycline resistance reservoirs. For final conclusions about the safety of these autochthonous cheeses regarding the possible tetracycline resistance transferability, the assessment of the entire cheese microbiota is needed.

Enterotoxin production and antimicrobial susceptibility in Staphylococci isolated from traditional raw milk cheeses in Serbia.

This study was undertaken to determine the prevalence of coagulase positive staphylococci (CPS) by examining a total of 71 raw milk cheeses. Additionally, enterotoxigenicity, antimicrobial susceptibility and the presence of mecA and mecC genes in the staphylococcal isolates were investigated. The isolation and enumeration procedure of CPS followed the International Organization for Standardization (ISO) standard. The presumptive staphylococci were identified by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) using the VITEK MS system. VIDAS® Staph enterotoxin II assay was used for the detection of classical enterotoxins. Antimicrobial susceptibility testing (AST) was accomplished performing the disk diffusion method. All suspected methicillin resistant staphylococci were investigated for the presence of mecA and mecC genes by PCR assay. A high prevalence (87.32%) of CPS was detected in the cheeses at contamination levels up to 5.58 log CFU g-1. Among 47 staphylococcal isolates screened for enterotoxin production, only one isolate, identified as S. hyicus, was confirmed as being enterotoxigenic. Resistance to penicillin (63.70%) was the most common resistance among the tested Staphylococcus aureus isolates. The dominant phenotypic resistance patterns in coagulase negative staphylococci (CNS) were resistance to ofloxacin and fusidic acid. All CNS isolates were susceptible to the clinically important antibiotics clindamycin, chloramphenicol, gentamicin, linezolid, rifampicin and trimethoprim-sulfamethoxazole. The mecA and mecC genes were not detected. To the best of our knowledge, this is the first study concerning evaluation of the presence of methicillin resistant staphylococci (MRS) in dairy products in Serbia.

Antioxidative Activity of Thyme (Thymus vulgaris) and Oregano (Origanum vulgare) Essential Oils and Their Effect on Oxidative Stability of Minced Pork Packaged Under Vacuum and Modified Atmosphere.

The antioxidant stability of minced pork treated with thyme and oregano essential oils (EOs) was determined. Minced pork containing different concentrations (0%, 0.3%, 0.6%, or 0.9%) of thyme (TEO) or oregano essential oil (OEO) and packaged under vacuum or modified atmosphere (MAP) (30%O2 /50%CO2 /20%N2 ) was evaluated within 15 days of refrigeration (3 ± 1 °C) storage. EOs were examined for scavenging capacity toward 2,2-diphenyl-1-picrylhydrazyl, nitric oxide radicals, and hydroxyl, and inhibition of lipid peroxidation and ferric ion reducing antioxidant power (butylated hydroxytoluene was used as positive controls). The order of antioxidative effectiveness was as follows: butylated hydroxytoluene > OEO > TEO, with significant differences between agents (P < 0.05). Lipid oxidation in meat was determined by monitoring malondialdehyde (MDA) formation and lipolysis was assessed by measuring the acidity index immediately and after 3, 6, 9, 12, and 15 days of storage. EOs significantly (P < 0.05) increased the stability of minced pork with respect to lipid oxidation compared with the control, and the antioxidative effect was dose-dependent. Moreover, vacuum packaging resulted in mince with significantly lower oxidation and lipolysis levels than modified atmosphere packaged mince (P < 0.05). The results demonstrate that both EOs examined effectively reduced lipid oxidation in raw pork mince after 2 weeks' storage. PRACTICAL APPLICATION: The natural food preservatives market is growing rapidly, as is high demand for organic foods. These results are likely to be of interest to the scientists, researchers, and persons who work in the meat industry. Results and discussion can contribute to a better understanding of antioxidative properties of essential oils in food model. Furthermore, no study has reported the effect of these MAP on pork oxidative stability.

Survival of Salmonella spp. in minced meat packaged under vacuum and modified atmosphere.

The effect of different modified atmosphere packaging regimes on the behavior of Salmonella spp. on minced meat was studied. Minced meat was experimentally contaminated with a Salmonella spp. cocktail (S. Enteritidis, S. Typhimurium, S. Infantis and S. Arizonae), packaged under vacuum or modified atmosphere with initial headspaces containing 20%O2/50%CO2/30%N2 and 20%O2/30%CO2/50%N2) and stored at 3±1°C for 12 days. Samples were analyzed for Salmonella spp., viable and lactic acid bacteria count every third day. Salmonella spp. counts decreased during storage in all packaging types, with reductions of about 1.5logCFU/g. A significant difference (p<0.01) was noted between Salmonella spp. counts in meat packaged in vacuum and modified atmospheres, although there was no significant difference in Salmonella spp. count between meat packaged in 50%CO2, and meat packaged in 30%CO2. At the end of the study, there were significant differences (p<0.01; p<0.05) in total viable and lactic acid bacterial counts between meat packaged in vacuum and modified atmosphere, and the lowest counts were noted in meat packaged in modified atmosphere with 50%CO2.

Inhibition of Salmonella by thyme essential oil and its effect on microbiological and sensory properties of minced pork meat packaged under vacuum and modified atmosphere.

The antibacterial activity of thyme essential oil (TEO) was evaluated against four serovars of Salmonella (S. Enteritidis, S. Typhimurium, S. Montevideo and S. Infantis), experimentally inoculated (106CFU/g) in minced pork, which was treated with different concentrations of the TEO (0.3%, 0.6% and 0.9%) packaged under vacuum or MAP (30%O2/50%CO2/20% N2) and stored at 3±1°C for 15days. GC-MS analysis of the TEO was performed in order to determine composition, and the predominant constituent was thymol (50.48%), followed by p-cymene and linalool. The minimum inhibitory concentration was determined for each Salmonella serovar studied. Among the tested active compounds, thymol and carvacrol exhibited the greatest inhibitory effect followed by TEO, with minimum inhibitory concentrations of 320 to 640µg/ml. S. Enteritidis was the most sensitive serovar. During the storage period, Salmonella counts in pork were reduced by 1.69-4.05logCFU/g. The influence of TEO on Enterobacteriaceae, lactic acid bacteria and total viable count was determined in control mince with no added Salmonella. The most pronounced antibacterial effect was achieved by the combination MAP and 0.9% TEO. Although the antibacterial activities of all studied concentrations of TEO in pork were evident and significant (P<0.05), sensory analysis showed that 0.3% TEO was the most acceptable to trained panellists.

Survival of Salmonella spp. in minced meat packaged under vacuum and modified atmosphere

Abstract The effect of different modified atmosphere packaging regimes on the behavior of Salmonella spp. on minced meat was studied. Minced meat was experimentally contaminated with a Salmonella spp. cocktail (S. Enteritidis, S. Typhimurium, S. Infantis and S. Arizonae), packaged under vacuum or modified atmosphere with initial headspaces containing 20%O2/50%CO2/30%N2 and 20%O2/30%CO2/50%N2) and stored at 3 ± 1 °C for 12 days. Samples were analyzed for Salmonella spp., viable and lactic acid bacteria count every third day. Salmonella spp. counts decreased during storage in all packaging types, with reductions of about 1.5 log CFU/g. A significant difference (p < 0.01) was noted between Salmonella spp. counts in meat packaged in vacuum and modified atmospheres, although there was no significant difference in Salmonella spp. count between meat packaged in 50%CO2, and meat packaged in 30%CO2. At the end of the study, there were significant differences (p < 0.01; p < 0.05) in total viable and lactic acid bacterial counts between meat packaged in vacuum and modified atmosphere, and the lowest counts were noted in meat packaged in modified atmosphere with 50%CO2.