Biofilm Formation of Probiotic Saccharomyces cerevisiae var. boulardii on Glass Surface during Beer Bottle Ageing

While brewing probiotic beer using Saccharomyces cerevisiae var. boulardii, we noticed the yeast potentially makes biofilm in glass bottles as the bottles get hazy. In this study, S. cerevisiae var. boulardii CNCM I-745 was used as a starter culture to produce probiotic beer. We studied the biofilm parameters combined with FLO11 mRNA expression and used light and scanning electron microscopy to document biofilm formation and structure. Our results revealed that ageing the beer and maturing from a sugar-rich to a sugar-limited beer, along with the stress factors from the brewing process (pH reduction and produced metabolites), led to an increase in biofilm mass; however, the viable count remained relatively stable (approximately 7.1 log10 cells/mL). Biofilm S. boulardii cells showed significantly higher FLO11 mRNA expression in the exponential and stationary phase compared to the planktonic cells. This study, therefore, provides evidence that S. cerevisiae var. boulardii makes biofilm on glass surfaces during beer bottle ageing. The impact of complications caused by formed biofilms on returnable bottles emphasizes the significance of this study

Evaluation of the probiotic and postbiotic potential of lactic acid bacteria from artisanal dairy products against pathogens

Introduction: Probiotic and postbiotic potential of thirty-two strains of lactic acid bacteria (LAB), obtained earlier from artisanal dairy sources in Pakistan, have been investigated against major multi-drug resistant (MDR) and food borne pathogenic bacteria. Methodology: LAB strains were identified by 16S rRNA gene sequencing and their antibacterial activity was assessed by the microdilution method. Four LAB isolates, Weissella confusa PL6, Enterococcus faecium PL7, and Lactobacillus delbrueckii PL11 and PL13 were shortlisted. Their ability to degrade lactose and safety for human consumption in terms of hemolysis and antibiotic susceptibility were assessed in vitro. The antibacterial components in the cell-free supernatants (CFSs) of isolate cultures were characterized biochemically by HPLC. Results: Acid neutralization but not protease treatment abolished the antibacterial activity of CFSs. Lactic, acetic and propionic acids were the main acids in the CFSs, and acid production peaked in the stationary phase of growth. The antibacterial activity of the LAB cultures resulted from secretion of organic acids that lowered the pH. The strains exhibited variable ability to degrade lactose and were non-hemolytic and susceptible to the most common antibiotics. Conclusions: These LAB strains are probiotic candidates for further investigation of their postbiotic role in naturally preserving processed foods and for attenuation of lactose intolerance.Peer reviewe

Lacticaseibacillus rhamnosus GG Survival and Quality Parameters in Kefir Produced from Kefir Grains and Natural Kefir Starter Culture

The study aimed to determine the effect of starter cultures (kefir grains and natural kefir starter culture without grains) on Lacticaseibacillus rhamnosus GG (LGG) survival and on the quality characteristics of kefir. To this end, the viability of probiotic L. rhamnosus GG strain and the rheological properties and quality parameters of kefir beverages were tested during storage over 21 days at 4 °C. The final LGG counts were 7.71 and 7.55 log cfu/mL in natural kefir starter culture and kefir grain, respectively. When prepared with probiotic bacteria, the syneresis values of kefir prepared using natural kefir starter culture was significantly lower (p 0.05). Moreover, all samples showed shear-thinning behavior. The flavor scores for kefir prepared using natural kefir starter culture were significantly higher than for the other samples (p 0.05). Overall, the results indicate that natural kefir starter culture could be a potential probiotic carrier

Pseudomonas fluorescens group bacterial strains interact differently with pathogens during dual-species biofilm formation on stainless steel surfaces in milk

In order to develop strategies for preventing biofilm formation in the dairy industry, a deeper understanding of the interaction between different species during biofilm formation is necessary. Bacterial strains of the P. fluorescens group are known as the most important biofilm-formers on the surface of dairy processing equipment that may attract and/or shelter other spoilage or pathogenic bacteria. The present study used different strains of the P. fluorescens group as background microbiota of milk, and evaluated their interaction with Staphylococcus aureus, Bacillus cereus, Escherichia coli O157:H7, and Salmonella Typhimurium during dual-species biofilm formation on stainless steel surfaces. Two separate scenarios for dual-species biofilms were considered: concurrent inoculation of Pseudomonas and pathogen (CI), and delayed inoculation of pathogen to the pre-formed Pseudomonas biofilm (DI). The gram-positive pathogens used in this study did not form dual-species biofilms with P. fluorescens strains unless they were simultaneously inoculated with Pseudomonas strains. E. coli O157:H7 was able to form dual-species biofilms with all seven P. fluorescens group strains, both in concurrent (CI) and delayed (DI) inoculation. However, the percentage of contribution varied depending on the P. fluorescens strains and the inoculation scenario. S. Typhimurium contributed to biofilm formation with all seven P. fluorescens group strains under the CI scenario, with varying degrees of contribution. However, under the DI scenario, S. Typhimurium did not contribute to the biofilm formed by three of the seven P. fluorescens group strains. Overall, these are the first results to illustrate that the strains within the P. fluorescens group have significant differences in the formation of mono-or dual-species biofilms with pathogenic bacteria. Furthermore, the possibility of forming dual-species biofilms with pathogens depends on whether the pathogens form the biofilm simultaneously with the P. fluorescens group strains or whether these strains have already formed a biofilm.Peer reviewe

Lacticaseibacillus rhamnosus GG Survival and Quality Parameters in Kefir Produced from Kefir Grains and Natural Kefir Starter Culture

The study aimed to determine the effect of starter cultures (kefir grains and natural kefir starter culture without grains) on Lacticaseibacillus rhamnosus GG (LGG) survival and on the quality characteristics of kefir. To this end, the viability of probiotic L. rhamnosus GG strain and the rheological properties and quality parameters of kefir beverages were tested during storage over 21 days at 4 °C. The final LGG counts were 7.71 and 7.55 log cfu/mL in natural kefir starter culture and kefir grain, respectively. When prepared with probiotic bacteria, the syneresis values of kefir prepared using natural kefir starter culture was significantly lower (p 0.05). Moreover, all samples showed shear-thinning behavior. The flavor scores for kefir prepared using natural kefir starter culture were significantly higher than for the other samples (p 0.05). Overall, the results indicate that natural kefir starter culture could be a potential probiotic carrier

Innovative approaches to nisin production

Nisin is a bacteriocin produced by Lactococcus lactis that has been approved by the Food Drug Administration for utilization as a GRAS status food additive. Nisin can inhibit spore germination and demonstrates antimicrobial activity against Listeria, Clostridium, Staphylococcus, and Bacillus species. Under some circumstances, it plays an immune modulator role and has a selective cytotoxic effect against cancer cells, although it is notable that the high production cost of nisin-a result of the low nisin production yield of producer strains-is an important factor restricting intensive use. In recent years, production of nisin has been significantly improved through genetic modifications to nisin producer strains and through innovative applications in the fermentation process. Recently, 15,400 IU ml-1 nisin production has been achieved in L. lactis cells following genetic modifications by eliminating the factors that negatively affect nisin biosynthesis or by increasing the cell density of the producing strains in the fermentation medium. In this review, innovative approaches related to cell and fermentation systems aimed at increasing nisin production are discussed and interpreted, with a view to increasing industrial nisin production.Peer reviewe

Comparative antioxidant potential of kefir and yogurt of bovine and non-bovine origins

The aim of this study was to compare the antioxidant potential of the yogurt and kefir produced from ewe, camel, goat, and cow milk. The antioxidant activity of the samples was assessed by measuring total phenolic content (TPC), 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activity, ferric reducing antioxidant power (FRAP) and 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) radical reducing capacity during 20-day storage at 4 oC. Kefir and yogurt prepared from ewe and camel milk had significantly higher antioxidative potential than samples made from goat and cow milk (P <0.05). Ewe kefir (74.55-80.11 mg GAE 100 mL(-1)) showed the highest TPC followed by cow kefir (65-73.15 mg GAE 100 mL(-1)), camel kefir (61.2-69.91 mg GAE 100 mL(-1)) and goat kefir (58.31-73.5 mg GAE 100 mL(-1)) (P <0.05). Camel yogurt possesses the highest TPC (56.5-68.25 mg GAE 100 mL(-1)) followed by ewe (40.32-46.5 mg GAE 100 mL(-1)), cow (29.5-35.5 mg GAE 100 mL(-1)) and goat (20.03-26.85 mg GAE 100 mL(-1)) yogurt (P <0.05). According to DPPH, FRAP, and ABTS results, the antioxidant activity of samples was as follows in descending order: ewe kefir, camel kefir, ewe yogurt, camel yogurt, cow kefir, goat kefir, goat yogurt, cow yogurt.Peer reviewe

Enhanced heterologous protein productivity by genome reduction in Lactococcus lactis NZ9000

Background: The implementation of novel chassis organisms to be used as microbial cell factories in industrial applications is an intensive research field. Lactococcus lactis, which is one of the most extensively studied model organisms, exhibits superior ability to be used as engineered host for fermentation of desirable products. However, few studies have reported about genome reduction of L. lactis as a clean background for functional genomic studies and a model chassis for desirable product fermentation. Results: Four large nonessential DNA regions accounting for 2.83% in L. lactis NZ9000 (L. lactis 9 k) genome (2,530,294 bp) were deleted using the Cre-loxP deletion system as the first steps toward a minimized genome in this study. The mutants were compared with the parental strain in several physiological traits and evaluated as microbial cell factories for heterologous protein production (intracellular and secretory expression) with the red fluorescent protein (RFP) and the bacteriocin leucocin C (LecC) as reporters. The four mutants grew faster, yielded enhanced biomass, achieved increased adenosine triphosphate content, and diminished maintenance demands compared with the wild strain in the two media tested. In particular, L. lactis 9 k-4 with the largest deletion was identified as the optimum candidate host for recombinant protein production. With nisin induction, not only the transcriptional efficiency but also the production levels of the expressed reporters were approximately three-to fourfold improved compared with the wild strain. The expression of lecC gene controlled with strong constitutive promoters P5 and P8 in L. lactis 9 k-4 was also improved significantly. Conclusions: The genome-streamlined L. lactis 9 k-4 outcompeted the parental strain in several physiological traits assessed. Moreover, L. lactis 9 k-4 exhibited good properties as platform organism for protein production. In future works, the genome of L. lactis will be maximally reduced by using our specific design to provide an even more clean background for functional genomics studies than L. lactis 9 k-4 constructed in this study. Furthermore, an improved background will be potentially available for use in biotechology.Peer reviewe

Antifungal metabolites, their novel sources, and targets to combat drug resistance

Excessive antibiotic prescriptions as well as their misuse in agriculture are the main causes of antimicrobial resistance which poses a growing threat to public health. It necessitates the search for novel chemicals to combat drug resistance. Since ancient times, naturally occurring medicines have been employed and the enormous variety of bioactive chemicals found in nature has long served as an inspiration for researchers looking for possible therapeutics. Secondary metabolites from microorganisms, particularly those from actinomycetes, have made it incredibly easy to find new molecules. Different actinomycetes species account for more than 70% of naturally generated antibiotics currently used in medicine, and they also produce a variety of secondary metabolites, including pigments, enzymes, and anti-inflammatory compounds. They continue to be a crucial source of fresh chemical diversity and a crucial component of drug discovery. This review summarizes some uncommon sources of antifungal metabolites and highlights the importance of further research on these unusual habitats as a source of novel antimicrobial molecules.Peer reviewe