Antimicrobial Compounds in Wine.

Ipsum vinum est potestas et possession (wine itself is power and possession). Wine is a complex system that triggers multisensory cognitive stimuli. Wine and its consumption are thoroughly intertwined with the development of human society. The beverage was appreciated in many ancient mythologies and plays an essential part in Christianity and rituals to this day. Wine has been said to enlighten and inspire artists and has even been prohibited by law and some religions, but has nevertheless played a role in human civilizations since the beginning. Winemaking is also a prospering and economically important industry and a longtime symbol of status and luxury. In winemaking, the formation of the final product is influenced by several factors that contribute to the chemical and sensory complexity often associated with quality vintages. Factors such as terroir, climatic conditions, variety of the grape, all aspects of the winemaking process to the smallest details, including metabolic processes carried out by yeast and malolactic bacteria, and the conditions for the maturation and storage of the final product, up to, and even beyond the point of deciding to open the bottle and enjoy the wine. In conjunction with the empiric and scientific process of winemaking, different molecules with antibacterial activity can be identified in wine during the production process, and several of them are clearly present in the final product. Some of these antibacterial components are phytochemicals, such as flavonoids and phenolic compounds, that may be delivered to the final product (wine) as a part of the grape, a variety of potential additive compounds, or from the oak barrels or clay amphoras used during the maturation process. Others are produced by yeasts and malolactic bacteria and play a role not only in the moderation of the fermentation process but contributing to the microbiological safety and beneficial properties spectra of the final product. Lactic acid bacteria, responsible for conducting malolactic fermentation, contribute to the final balance of the wine but are also directly involved in the production of different compounds exhibiting antibacterial activity. Some examples of these compounds include bacteriocins (antibacterial peptides), diacetyl, organic acids, reuterin, hydrogen peroxide, and carbon dioxide. Major aspects of these different beneficial metabolites are the subject of discussion in this review with the aim of highlighting their beneficial functions.

Plasmid-Associated Bacteriocin Produced by Pediococcus pentosaceus Isolated from Smoked Salmon: Partial Characterization and Some Aspects of his Mode of Action.

Strain ST3Ha, isolated from commercially available smoked salmon, was identified as Pediococcus pentosaceus based on biochemical and physiological tests and 16S rRNA sequencing. Strain ST3Ha produces a class IIa bacteriocin active against lactic acid bacteria, Listeria monocytogenes and Enterococcus faecalis. The antimicrobial peptide was inactivated by proteolytic enzymes, confirming his proteinaceous nature, but was not affected when treated with α-amylase, SDS, Tween 20, Tween 80, urea, and EDTA. No change in activity was recorded after 2 h at pH values between 2.0 and 9.0 and after treatment at 100 °C for 120 min or 121 °C for 15 min. The mode of action against Listeria ivanovii subsp. ivanovii ATCC 19119 and E. faecalis ATCC 19443 was bactericidal, resulting in cell lyses and enzyme leakage. The highest level of activity (1.6 × 106 AU/mL) was recorded when cells were grown at 37 °C or 30 °C in MRS broth (pH 6.5). Antimicrobial peptide ST3Ha adsorbs at high levels to the sensitive test organisms on strain-specific manner and depending on incubation temperature, environmental pH, and presence of supplemented chemicals. Based on PCR analysis, P. pentosaceus ST3Ha harbor a 1044-bp plasmid-associated fragment corresponding in size to that recorded for pediocin PA-1. Sequencing of the fragment revealed a gene identical to pedB, reported for pediocin PA-1. The combined application of the low levels (below MIC) of ciprofloxacin and bacteriocin ST3Ha results in the synergetic effect in the inhibition of L. ivanovii subsp. ivanovii ATCC 19119. Expressed by P. pentosaceus ST3Ha, bacteriocin was characterized as low cytotoxic, a characteristic relevant for its application in food industry and/or in human and veterinary medical practices.

Beneficial features of pediococcus: from starter cultures and inhibitory activities to probiotic benefits.

Pediococci are lactic acid bacteria (LAB) which have been used for centuries in the production of traditional fermented foods. There fermentative abilities were explored by the modern food processing industry in use of pediococci as starter cultures, enabling the production of fermented foods with distinct characteristics. Furthermore, some pediococci strains can produce bacteriocins and other antimicrobial metabolites (AMM), such as pediocins, which are increasingly being explored as bio-preservatives in various food matrices. Due to their versatility and inhibitory spectrum, pediococci bacteriocins and AMM are being extensively researched not only in the food industry, but also in veterinary and human medicine. Some of the pediococci were evaluated as potential probiotics with different beneficial areas of application associated with human and other animals' health. The main taxonomic characteristics of pediococci species are presented here, as well as and their potential roles and applications as starter cultures, as bio-preservatives and as probiotic candidates.

Antimicrobial properties of Pediococcus acidilactici and Pediococcus pentosaceus isolated from silage.

AIMS: The objective of this study was to isolate multifunctional bacteriocin-producing strains; to characterize the expressed bacteriocin for the control of Listeria monocytogenes and vancomycin-resistant Enterococcus; to evaluate the safety of studied strains; and to explore their antifungal activity. METHODS AND RESULTS: Two Pediococcus strains were isolated from silage samples obtained from an organic farm in Belogradchik, Bulgaria. The strains were identified by 16S rRNA sequencing analysis and characterized as bacteriocins producers. Strong antimicrobial activity was detected against more than 74 different strains of Listeria monocytogenes, 27 different vancomycin-resistant Enterococcus strains. In addition, studied strains were able to inhibit the growth of strains of Alternaria alternate, Aspergillus flavus, Aspergillus niger, Cladosporium sphaerospermum, Penicillium chrysogenum and Penicillium expansum. Some aspects of the antimicrobial mode of action were evaluated, including killing curves and aggregation properties. Both strains generated positive PCR results for the presence of pediocin PA-1, but not for other bacteriocins evaluated in this screening process. Metabolomic analysis of the cell-free supernatants from both strains was performed in order to explain the observed antifungal activity against different moulds. According to PCA and PLS-DA score plot, P. acidilactici ST3522BG and P. pentosaceus ST3633BG were clearly clustered from control (MRS). Increases in the production of benzoic acid, 2-hydroxyisocaproic acid, ß-phenyl-lactic acid, α-hydroxybutyric acid and 1,3-butanediol were recorded, these metabolites were previously described as antifungal. CONCLUSIONS: Pediococcus acidilactici ST3522BG and P. pentosaceus ST3633BG were evaluated as producing bacteriocin strains with high specificity against Listeria and vancomycin-resistant Enterococcus species. In addition, both investigated Pediococcus strains were evaluated as producer of effective antifungal metabolites with potential for the inhibition of mycotoxin-producing moulds. SIGNIFICANCE AND IMPACT OF THE STUDY: To the best of our knowledge, this report is a pioneer in the evaluation of Pediococcus strains isolated from silage with highly specific bacteriocinogenic antimicrobial activity against Listeria spp. and vancomycin-resistant Enterococcus spp., and antifungal activity against mycotoxin-producing moulds.

Bacillus spore-forming probiotics: benefits with concerns?

Representatives of the genus Bacillus are multifunctional microorganisms with a broad range of applications in both traditional fermentation and modern biotechnological processes. Bacillus spp. has several beneficial properties. They serve as starter cultures for various traditional fermented foods and are important biotechnological producers of enzymes, antibiotics, and bioactive peptides. They are also used as probiotics for humans, in veterinary medicine, and as feed additives for animals of agricultural importance. The beneficial effects of bacilli are well-reported and broadly acknowledged. However, with a better understanding of their positive role, many questions have been raised regarding their safety and the relevance of spore formation in the practical application of this group of microorganisms. What is the role of Bacillus spp. in the human microbial consortium? When and why did they start colonizing the gastrointestinal tract (GIT) of humans and other animals? Can spore-forming probiotics be considered as truly beneficial organisms, or should they still be approached with caution and regarded as "benefits with concerns"? In this review, we not only hope to answer the above questions but to expand the scope of the conversation surrounding bacilli probiotics.

Could Probiotics and Postbiotics Function as "Silver Bullet" in the Post-COVID-19 Era?

We are currently experiencing the realities of the most severe pandemic within living memory, with major impacts on the health and economic well-being of our planet. The scientific community has demonstrated an unprecedented mobilization capability, with the rapid development of vaccines and drugs targeting the protection of human life and palliative measures for infected individuals. However, are we adequately prepared for ongoing defense against COVID-19 and its variants in the post-pandemic world? Moreover, are we equipped to provide a satisfactory quality of life for individuals who are recovering from COVID-19 disease? What are the possibilities for the acceleration of the recovery process? Here, we give special consideration to the potential and already-demonstrated role of probiotics and traditional medical approaches to the management of current and potential future encounters with our major virus adversaries.

Technology and safety assessment for lactic acid bacteria isolated from traditional Bulgarian fermented meat product "lukanka"

Abstract The present work discusses the technological and new selection criteria that should be included for selecting lactic acid bacteria for production of fermented meat. Lactic acid bacteria isolated from Bulgarian traditional fermented "lulanka" salami was studied regarding some positive technological parameters (growth at different temperature, pH, and proteolytic activity). The presence of genes related to the virulence factors, production of biogenic amines, and vancomycin resistance were presented in low frequency in the studied lactic acid bacteria. On the other hand, production of antimicrobial peptides and high spread of bacteriocin genes were broadly presented. Very strong activity against L. monocytogenes was detected in some of the studied lactic acid bacteria. In addition, the studied strains did not present any antimicrobial activity against tested closely related bacteria such as Lactobacillus spp., Lactococcus spp., Enterococcus spp. or Pediococcus spp. To our knowledge this is the first study on the safety and antimicrobial properties of lactic acid bacteria isolated from Bulgarian lukanka obtained by spontaneous fermentation.

Technology and safety assessment for lactic acid bacteria isolated from traditional Bulgarian fermented meat product "lukanka".

The present work discusses the technological and new selection criteria that should be included for selecting lactic acid bacteria for production of fermented meat. Lactic acid bacteria isolated from Bulgarian traditional fermented "lulanka" salami was studied regarding some positive technological parameters (growth at different temperature, pH, and proteolytic activity). The presence of genes related to the virulence factors, production of biogenic amines, and vancomycin resistance were presented in low frequency in the studied lactic acid bacteria. On the other hand, production of antimicrobial peptides and high spread of bacteriocin genes were broadly presented. Very strong activity against L. monocytogenes was detected in some of the studied lactic acid bacteria. In addition, the studied strains did not present any antimicrobial activity against tested closely related bacteria such as Lactobacillus spp., Lactococcus spp., Enterococcus spp. or Pediococcus spp. To our knowledge this is the first study on the safety and antimicrobial properties of lactic acid bacteria isolated from Bulgarian lukanka obtained by spontaneous fermentation.

High-level expression, purification and characterisation of porcine ß-defensin 2 in Pichia pastoris and its potential as a cost-efficient growth promoter in porcine feed.

Porcine ß-defensin 2 (pBD2), a recently discovered porcine defensin that is produced by the intestine, exerts antimicrobial activities and innate immune effects that are linked to intestinal diseases in pigs. Here, we report a codon-optimised protein corresponding to mature pBD2 cDNA that was expressed and purified in Pichia pastoris yeast. The highest amount of secreted protein (3,694.0 mg/L) was reached 144 h into a 150-h induction during high-density cultivation. Precipitation followed by gel exclusion chromatography yielded 383.7 mg/L purified recombinant pBD2 (rpBD2) with a purity of ~93.7 %. Two recombinant proteins of 5,458.5 and 5,258.4 Da were detected in the mass spectrum due to variation in the amino-terminus. The rpBD2 exhibited high antimicrobial activity against a broad range of pig pathogenic bacteria (minimal inhibitory concentration [MIC] 32-128 µg/mL); the highest activity was observed against Salmonella choleraesuis, Staphylococcus aureus and Streptococcus suis (MIC 32-64 µg/mL). However, rpBD2 also inhibited the growth of probiotics such as Lactobacillus plantarum, Bacillus subtilis and Saccharomyces cerevisiae, but at lower efficacies than the pathogens. Purified or unpurified rpBD2 also maintained high activity over a wide range of pH values (2.0-10.0), a high thermal stability at 100 °C for 40 min and significant resistance to papain, pepsin and trypsin. In addition, the activity of rpBD2 towards S. aureus was unaffected by 10 mM dithiothreitol (DTT) and 20 % dimethyl sulphoxide (DMSO). Our results suggest that pBD2 could be produced efficiently in large quantities in P. pastoris and be a substitute for traditional antibiotics for growth promotion in the porcine industry.