Jolanta Godziszewska

MobC of conjugative RA3 plasmid from IncU group autoregulates the expression of bicistronic mobC-nic operon and stimulates conjugative transfer.

BackgroundThe IncU conjugative transfer module represents highly efficient promiscuous system widespread among conjugative plasmids of different incompatibility groups. Despite its frequent occurrence the mechanisms of relaxosome formation/action are far from understood. Here we analyzed the putative transfer auxiliary protein MobC of the conjugative plasmid RA3 from the IncU incompatibility group.ResultsMobC is a protein of 176 amino acids encoded in the bicistronic operon mobC-nic adjacent to oriT. MobC is homologous to prokaryotic transcription factors of the ribbon-helix-helix (RHH) superfamily. Conserved LxxugxNlNQiaxxLn motif clusters MobC with the clade of conjugative transfer auxilliary proteins of MobP relaxases. MobC forms dimers in solution and autoregulates the expression of mobCp by binding to an imperfect palindromic sequence (OM) located between putative -35 and -10 motifs of the promoter. Medium-copy number test plasmid containing the oriT-mobCp region is mobilized with a high frequency by the RA3 conjugative system. The mutations introduced into OM that abolished MobC binding in vitro decreased 2-3 fold the frequency of mobilization of the test plasmids. The deletion of OM within the RA3 conjugative module had no effect on transfer if the mobC-nic operon was expressed from the heterologous promoter. If only nic was expressed from the heterologous promoter (no mobC) the conjugative transfer frequency of such plasmid was 1000-fold lower.ConclusionThe MobC is an auxiliary transfer protein of dual function. It autoregulates the expression of mobC-nic operon while its presence significantly stimulates transfer efficiency.

Oxidative and microbial stability of poultry meatballs added with coriander extracts and packed in cold modified atmosphere

ABSTRACT The aim of the study was to determine the effects of adding of coriander extracts to turkey meatballs. Meat samples were stored in modified atmosphere at 4°C ± 1°C for 9 days. The addition of coriander extract at 500 ppm level delayed the process of lipid oxidation for the period of 6 days of storage and growth inhibition of aerobic microorganisms for the period of 9 days of storage. The usage of a dose of 500 ppm as well as a dose of 200 ppm had no significant effect on the sensory features, but had an impact on colour parameters of the meatballs. In the groups with coriander extract, volatile terpenes were identified.

Mobile antibiotic resistance - the spread of genes determining the resistance of bacteria through food products.

In recent years, more and more antibiotics have become ineffective in the treatment of bacterial nfections. The acquisition of antibiotic resistance by bacteria is associated with circulation of genes in the environment. Determinants of antibiotic resistance may be transferred to pathogenic bacteria. It has been shown that conjugation is one of the key mechanisms responsible for spread of antibiotic resistance genes, which is highly efficient and allows the barrier to restrictions and modifications to be avoided. Some conjugative modules enable the transfer of plasmids even between phylogenetically distant bacterial species. Many scientific reports indicate that food is one of the main reservoirs of these genes. Antibiotic resistance genes have been identified in meat products, milk, fruits and vegetables. The reason for such a wide spread of antibiotic resistance genes is the overuse of antibiotics by breeders of plants and animals, as well as by horizontal gene transfer. It was shown, that resistance determinants located on mobile genetic elements, which are isolated from food products, can easily be transferred to another niche. The antibiotic resistance genes have been in the environment for 30 000 years. Their removal from food products is not possible, but the risks associated with the emergence of multiresistant pathogenic strains are very large. The only option is to control the emergence, selection and spread of these genes. Therefore measures are sought to prevent horizontal transfer of genes. Promising concepts involve the combination of developmental biology, evolution and ecology in the fight against the spread of antibiotic resistance.

Concerted action of NIC relaxase and auxiliary protein MobC in RA3 plasmid conjugation.

Conjugative transfer of the broad‐host‐range RA3 plasmid, the archetype of the IncU group, relies on the relaxase NIC that belongs to the as yet uncharacterized MOBP4 subfamily. NIC contains the signature motifs of HUH relaxases involved in Tyr nucleophilic attack. However, it differs in the residue involved in His activation for cation coordination and was shown here to have altered divalent cation requirements. NIC is encoded in the mobC‐nic operon preceded directly by oriT, where mobC encodes an auxiliary transfer protein with a dual function: autorepressor and stimulator of conjugative transfer. Here an interplay between MobC and NIC was demonstrated. MobC is required for efficient NIC cleavage of oriT in supercoiled DNA whereas NIC assists MobC in repression of the mobC‐nic operon. A 7‐bp arm of IR3 (IR3a) was identified as the binding site for NIC and the crucial nucleotides in IR3a for NIC recognition were defined. Fully active oriTRA3 was delineated to a 47‐bp DNA segment encompassing a conserved cleavage site sequence, the NIC binding site IR3a and the MobC binding site OM. This highly efficient RA3 conjugative system with defined requirements for minimal oriT could find ample applications in biotechnology and computational biology where simple conjugative systems are needed.

The Effect of PUFA-Rich Plant Oils and Bioactive Compounds Supplementation in Pig Diet on Color Parameters and Myoglobin Status in Long-Frozen Pork Meat

The study evaluated the effect of pig diet supplementation with rapeseed or linseed oil, and vitamin E or selenium, or both vitamin E and selenium on color parameters and myoglobin content of pork Semimembranosus muscle after long-term freezing storage during nine months. The influence of the type of the bioactive compounds added to pig diet on the content of myoglobin or oxymyoglobin, metmyoglobin and deoksymyoglobin in Semimembranosus m. was also assessed. The results indicate that the presence of oils rich in polyunsaturated fatty acids (PUFA) in pig diet improves the color of pork meat. Supplementation of dietary plant oils or dietary oils with antioxidants tended to increase significantly the concentration of oxymyoglobin and decrease the concentration of metmyoglobin in meat compared to the control group. The highest content of oxymyoglobin was observed in meat obtained from pigs fed diets with linseed oil. The best color scores (highest a* parameter) was noted for rapeseed oil group (with no addition of antioxidants). In conclusion, the addition of antioxidants to pigs’ forage supplemented with PUFA-rich oils is not recommended in order to improve color of long-term frozen pork.

Antioxidant effect of sage (Salvia officinalis L.) extract on turkey meatballs packed in cold modified atmosphere

ABSTRACT The aim of the study was to determine the influence of sage extracts (0.02% and 0.05%) on the quality of turkey meatballs packed in a modified atmosphere (80% O2; 20% CO2) and stored at 4°C over a period of 9 days. Sage extract added to meatballs had a high antioxidant capacity due to a phenolic compounds: 47.92 mg of gallic acid (GAE)/100 g dw including high flavonoids contents (20.47 mg GAE/100 g dw) and phenolic acids (8.14 GAE/100 g dw) contents. During the storage, lipid oxidation and microbial growth increased rapidly and the results showed that the influence of sage extract on meatballs was the most noticeable on the last day of storage. Furthermore, analysis of the volatile compounds indicated that the addition of sage extract delayed formation of lipid-derived products of oxidation through storage of the turkey meatballs. Sage extract has a potential as a natural preservative in the meat industry.

The sensory quality of allergen-controlled, fat-reduced, salt-reduced pork-ostrich sausages during storage

BACKGROUND New meat products tailored to consumer health should be characterised by reduced sodium, fat and cholesterol contents and other health-promoting benefits. However, the food sectors greatest challenge is allergen-free production. Consumers are not willing to compromise the sensory quality of meat products for health. The aim of the present study was to analyse the influence of the storage time on the physical properties and consumer acceptance of allergen-controlled, fat-reduced, salt-reduced pork-ostrich sausages. The study focused on pork-ostrich sausages produced in accordance with a new patented technology, which focused on eliminating cross-contamination on-line in the plant, eliminating cross-contamination after preparation, and eliminating spices with high allergy potential. The production was focused on reducing fat (by approximately 50%) and salt (by approximately 30%) levels. RESULTS No changes in the texture parameters of the sausage were observed during storage time; however, some changes in colour were observed. There were no significant differences in sensory consumer acceptability of pork-ostrich sausage after 14 days of storage; thus, it may be stated that the instrumentally assessed differences in colour did not influence consumer acceptance. CONCLUSION The applied fat and NaCl reduction in the pork-ostrich sausages contributed to high consumer ratings and was not correlated with saltiness acceptability.