Advances in the applications of Bacteriophages and phage products against food-contaminating bacteria.

Food-contaminating bacteria pose a threat to food safety and the economy by causing foodborne illnesses and spoilage. Bacteriophages, a group of viruses that infect only bacteria, have the potential to control bacteria throughout the "farm-to-fork continuum". Phage application offers several advantages, including targeted action against specific bacterial strains and minimal impact on the natural microflora of food. This review covers multiple aspects of bacteriophages applications in the food industry, including their use as biocontrol and biopreservation agents to fight over 20 different genera of food-contaminating bacteria, reduce cross-contamination and the risk of foodborne diseases, and also to prolong shelf life and preserve freshness. The review also highlights the benefits of using bacteriophages in bioprocesses to selectively inhibit undesirable bacteria, such as substrate competitors and toxin producers, which is particularly valuable in complex microbial bioprocesses where physical or chemical methods become inadequate. Furthermore, the review briefly discusses other uses of bacteriophages in the food industry, such as sanitizing food processing environments and detecting specific bacteria in food products. The review also explores strategies to enhance the effectiveness of phages, such as employing multi-phage cocktails, encapsulated phages, phage products, and synergistic hurdle approaches by combining them with antimicrobials.

[Rapid identification of Ralstonia pickettii using PCR-nucleic acid test strips].

OBJECTIVE: To develop a method for rapid detection of Ralstonia pickettii in water for pharmaceutical purpose using PCR-nucleic acid test strips. METHODS: The genomic DNA of Ralstonia pickettii was extracted by boiling method. A pair of specific primers targeting the 16S rDNA with FITC and biotin labeling of the 5' ends was designed and cloned into competent E. coli DH5α cells. The nucleic acid test strips were assembled, and the workload of streptavidin labeled with colloidal gold and antibody concentration in the reaction system was optimized. After verification of the reaction mechanism and assessment of the test sensitivity, specificity and stability, the test strip was used for detecting 7 known strains of Ralstonia pickettii detected in pharmaceutical water, and an evolutionary tree was constructed to analyze the source of contamination. RESULTS: The genomic DNA extracted by boiling method had a purity between 1.8 and 2.0, and the PCR products showed a 100% similarity of with Ralstonia pickettii 16S rDNA registered in GenBank. Using the colloidal gold amplification principle, in every 100 µL colloidal gold solution, 3.5 µL streptavidin was added; the detection line on nitrocellulose membrane was 2.0 mg·mL-1 anti FITC antibody, and the quality control line was 1.2 mg · mL-1 biotinylated BSA, and they generate a red band after binding with positive amplification product. Specificity test of the assembled test strip yielded consistent result with agarose gel electrophoresis without cross reaction with Acinetobacter, Aeromonas, Pseudomonas, or Leclercia adecarboxylata. Sensitivity test of the strip showed a lower detection limit for DNA concentration of 10-5 ng/µL, with a sensitivity 1000 times that of agarose gel electrophoresis. The test strip still had good performance after storage for 3, 6, 9 and 12 months. CONCLUSION: We successfully developed a PCR-nucleic acid test strip for convenient and cost-effective detection of Ralstonia pickettii with good specificity and sensitivity and low cost to facilitate daily monitoring of pharmaceutical water contaminations.

Removal of bacteria from stallion semen by colloid centrifugation.

Bacteria (environmental contaminants and occasionally potential pathogens) are found in most stallion ejaculates and may negatively affect sperm quality during storage. Since the use of antibiotics can lead to the development of resistance, an alternative means of microbial control is desirable. The removal of bacteria from stallion semen using Single Layer Centrifugation through Androcoll-E was investigated. Known doses of cultured bacteria were added to freshly collected ejaculates (15mL aliquots) before processing by Single Layer Centrifugation. The resulting sperm pellets and controls (not processed by Single Layer Centrifugation) were cultured and the bacteria identified. In experiment 1, doses of E. coli from 2×10(2) to 2×10(7) colony forming units were added to aliquots of semen. In experiment 2, Taylorella equigenitalis or a mix of E. coli, Klebsiella pneumoniae and Streptococcus equi subsp. zooepidemicus (approximately 7×10(6), 5×10(6), and 6×10(6)cfu, respectively) were added to 15mL aliquots of semen. In experiment 1, more than 90% of the bacteria were removed where loading doses were >×10(4)cfu/mL. In experiment 2, varying proportions of different bacteria were removed, ranging from 68% for naturally occurring Corynebacterium spp. to >97% for added cultured E. coli. Thus, Single Layer Centrifugation can separate spermatozoa from many, but not all bacteria in stallion ejaculates and could be a useful alternative to adding antibiotics to semen extenders to control bacterial contamination. However, further research is needed to determine the effect of small numbers of bacteria on sperm quality.

Effects of corn meal and sulphuric acid on the production of cachaça / Efeitos do fubá e ácido sulfúrico sobre a produção de cachaça

This work was carried out to evaluate the effects of using corn meal and treating yeasts with sulfuric acid on fermentation microorganisms, wine acidity, ethanol content and cachaça yield and composition. The experiment was arranged in randomized block design, in a 2x3 factorial with five replications. The methods applied in this study are recommended by distilleries. Results showed that the yeast sulfuric acid treatment transferred acidity to the fermenting juice, without any influence on yeast viability, ethanol content and cachaça yield. On the other hand, the acid treatment controlled lactic bacteria in the inoculum. Addition of corn meal increased the concentration of lactic bacteria in the end of the fermentation and increased the levels of higher alcohols in cachaça, especially propyl and isobutyl alcohol.

Avaliou-se o efeito da adição do fubá de milho no mosto de xarope de cana e o tratamento ácido do pé-de-cuba sobre a microbiota do processo fermentativo, acidez do vinho, grau alcoólico, rendimento e composição da cachaça. O delineamento experimental utilizado foi o de blocos casualizados, no esquema fatorial 2x3 e cinco repetições. A metodologia empregada e as análises foram as recomendadas pelo setor aguardenteiro. Os resultados permitiram concluir que a adição do ácido sulfúrico no pé-de-cuba transferiu a acidez para o vinho, não influenciando na viabilidade das leveduras, rendimento e composição da cachaça. Por outro lado, a acidificação do meio controlou as bactérias láticas no pé-de-cuba. A adição do fubá aumentou a concentração de bactérias lácticas ao final do processo fermentativo e dos álcoois homólogos superiores na cachaça, particularmente, os álcoois propílico e isobutílico.