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This study shows that silage, the product of acid autolysis, made from Anisakis-infested fish viscera can be safely incorporated into aquaculture feeds. Its use enables the recovery of large quantities of marine by-products, reduces allergenicity risks, maintains feed performance, and supports more sustainable, resource-efficient aquaculture. Anisakis simplex allergens are highly resistant to conventional processing, posing a health risk that limits the use of infested fish both for human consumption and for animal feed. To assess the safety and nutritional value of viscera silage, hake (Merluccius merluccius) viscera were processed with formic acid and stored for four months. Two experimental diets were prepared—one containing silage and another with untreated viscera—alongside a commercial control feed. Juvenile gilthead seabreams (Sparus aurata) were fed these diets for 86 days, and growth performance, muscle composition, and allergenicity were analysed. All groups achieved more than 20% weight gain, and the silage-fed fish showed the lowest residual allergenicity (15.3%), compared to the fish fed untreated viscera (24%) and commercial feed (21.9%). Silage processing caused extensive protein hydrolysis, reducing high-molecular-weight proteins, though some IgE-reactive fragments remained. Proteomic analysis detected only seabream structural and metabolic proteins, with no confirmed Anisakis allergens. These findings indicate that silage produced from Anisakis-infested viscera should not pose a higher risk to consumers than the fishmeal used in aquaculture feeds and could support the valorisation of currently discarded fish viscera.

Background Fresh milk and natural environmental conditions are used to produce traditional cheeses. Such cheeses are produced by dozens of different types of microbes. Non-starter lactobacilli are the most responsible genus of lactic acid bacteria exhibiting key technological and health promoting traits. The purpose of this study is to isolate Lactobacillus bacteria from conventional Egyptian cheeses and analyse their probiotic potential and technological properties. Results Lactobacillus isolates (33 isolates) were isolated from different Egyptian cheeses. Our results revealed that 18.18% of the isolates were fast-acidifying, 30.3% were medium-acidifying and 51.5% were slow-acidifying isolates. The results of autolytic activity showed that 24.3% of the isolates were good autolysis, 33.3% were fair autolysis, while 42.4% were poor autolysis. Fifteen isolates produced exopolysaccharides, while 9 isolates exhibited antimicrobial activities against Lactobacillus bulgaricus 340. All the isolates were resistant to pH 3 for 3 h except isolate No. 15 (MR4). The growth rate of the isolates ranged from 42.25 to 85.25% at 0.3% bile salts after 3 h of incubation. The surviving percentage of the Lactobacillus isolates decreased with increasing incubation time or the percentage of bile salts greater than 0.3%. All the isolates grew after incubation in artificial gastric and intestinal fluids. The auto-aggregation of 15 isolates ranged from 43.13 to 72.77%. Lacticaseibacillus paracasei BD3, Lactiplantibacillus plantarum BR4 and Limosilactobacillus fermentum MR2 were sensitive to the majority of the tested antibiotics and showed good bile salt hydrolase activity. Conclusion L. paracasei BD3, L. plantarum BR4 and L. fermentum MR2 were isolated from Egyptian cheeses and showed probiotic and technological characterization, which are valuable for their practical application as starters, adjunct and protective cultures in cheese making.

The cell wall of Gram-positive bacteria is a complex assemblage of glycopolymers and proteins. It consists of a thick peptidoglycan sacculus that surrounds the cytoplasmic membrane and that is decorated with teichoic acids, polysaccharides, and proteins. It plays a major role in bacterial physiology since it maintains cell shape and integrity during growth and division; in addition, it acts as the interface between the bacterium and its environment. Lactic acid bacteria (LAB) are traditionally and widely used to ferment food, and they are also the subject of more and more research because of their potential health-related benefits. It is now recognized that understanding the composition, structure, and properties of LAB cell walls is a crucial part of developing technological and health applications using these bacteria. In this review, we examine the different components of the Gram-positive cell wall: peptidoglycan, teichoic acids, polysaccharides, and proteins. We present recent findings regarding the structure and function of these complex compounds, results that have emerged thanks to the tandem development of structural analysis and whole genome sequencing. Although general structures and biosynthesis pathways are conserved among Gram-positive bacteria, studies have revealed that LAB cell walls demonstrate unique properties; these studies have yielded some notable, fundamental, and novel findings. Given the potential of this research to contribute to future applied strategies, in our discussion of the role played by cell wall components in LAB physiology, we pay special attention to the mechanisms controlling bacterial autolysis, bacterial sensitivity to bacteriophages and the mechanisms underlying interactions between probiotic bacteria and their hosts.

Spent brewer’s yeast is an excellent source of a variety of bioactive substances. In this study, for the first time, the focus was solely on investigating the influence of three industrially applicable cell disruption methods (cell mill, sonotrode, and autolysis) on selected substance groups relevant for physiology and process technology. A consistent spent yeast (Saccharomyces cerevisiae TUM 68) produced in a standardized industrial pilot top-fermenting process was used as a raw material. Using mechanical methods, i.e., cell mill and sonotrode, the protein content (as not hydrolyzed in free amino acids), the trehalose and the total fat content in the yeast extract were increased compared with those produced in the autolytic method. The analyzed B vitamin levels were also higher, the biologically active 5-CH3-H4folate in particular had the greatest proportion in the folate vitamer distribution of the mechanically produced yeast extracts. An increased level of non-fragmented genomic and mitochondrial DNA could also be found in the yeast extract produced via the mechanical methods. The antioxidative and reduction potential was decreased by the degradation of polyphenols and glutathione in the yeast extract following autolysis. The mineral, RNA, glycogen, glucose, fructose and ash contents did not differ significantly. Therefore, the cell mill and sonotrode offered a good alternative method to conventional autolytic procedures, especially to transfer physiologically relevant substance groups in higher concentrations to the yeast extract.

Sourdough is a type of dough fermented by yeast and lactic acid bacteria (LAB) used as sponge dough in bread making. Sourdough may have four classifications according to the fermentation type and the technological process used. On sourdough type I, the fermentation of yeasts and LAB present in the flour occurs spontaneously whereas in type II, fermentation occurs after the inoculation of a starter culture. Type III is simply type II sourdough dehydrated and type IV is a mixture of type I and type II sourdough, produced on laboratory scale. With LAB’s growth during fermentation, pH decreases with subsequent hydrolysis of starch and protein, favoring the growth of yeasts. The yeast in turn, releases amino acids during autolysis, contributing to the growth of LAB. Due to this synergistic growth of LAB and yeasts, the products obtained from sourdough show more concentration of flavor, more elastic dough, and a longer shelf life than bread fermented only by yeast. Although, the benefits of this technology, using sourdough, is present just in a few countries, where it is also possible to obtain the dehydrated sourdough starters, for domestic use, in local supermarkets. Due to its fermentative process complexity, this paper presents updated information about the fabrication process of sourdough, main factors affecting this process, benefits of sourdough use for bakery products, and the varieties of dehydrated sourdough commercially available throughout the world.

Outer Membrane Vesicles (OMVs) are ubiquitous in bacterial environments and enable interactions within and between species. OMVs are observed in lab-grown and environmental biofilms, but our understanding of their function comes primarily from planktonic studies. Planktonic OMVs assist in toxin delivery, cell-cell communication, horizontal gene transfer, small RNA trafficking, and immune system evasion. Previous studies reported differences in size and proteomic cargo between planktonic and agar plate biofilm OMVs, suggesting possible differences in function between OMV types. In Pseudomonas aeruginosa interstitial biofilms, extracellular vesicles were reported to arise through cell lysis, in contrast to planktonic OMV biogenesis that involves the Pseudomonas Quinolone Signal (PQS) without appreciable autolysis. Differences in biogenesis mechanism could provide a rationale for observed differences in OMV characteristics between systems. Using nanoparticle tracking, we found that P. aeruginosa PAO1 planktonic and biofilm OMVs had similar characteristics. However, P. aeruginosa PA14 OMVs were smaller, with planktonic OMVs also being smaller than their biofilm counterparts. Large differences in Staphylococcus killing ability were measured between OMVs from different strains, and a smaller within-strain difference was recorded between PA14 planktonic and biofilm OMVs. Across all conditions, the predatory ability of OMVs negatively correlated with their size. To address biogenesis mechanism, we analyzed vesicles from wild type and pqsA mutant biofilms. This showed that PQS is required for physiological-scale production of biofilm OMVs, and time-course analysis confirmed that PQS production precedes OMV production as it does in planktonic cultures. However, a small sub-population of vesicles was detected in pqsA mutant biofilms whose size distribution more resembled sonicated cell debris than wild type OMVs. These results support the idea that, while a small and unique population of vesicles in P. aeruginosa biofilms may result from cell lysis, the PQS-induced mechanism is required to generate the majority of OMVs produced by wild type communities.

The proteinogenic composition of yeast extract products varies from experience for manufacturing reasons, though also due to the yeast starting product. Therefore, this study is the first to focus solely on the influence of three industrially applicable cell disruption methods (cell mill, ultrasonic sonotrode and autolysis) on the amino acid and protein composition of a yeast extract. A consistent spent yeast (Saccharomyces cerevisiae TUM 68) produced in a standardized industrial pilot top-fermenting process was used as a raw material for the first time. The disruption effectiveness of autolysis (98%) was higher than that of the mechanical methods (80%), as well as the cleavage of amino acids from the cell protein (307, 155 and 115 mg per g yeast extract for autolysis, sonotrode and cell mill). The proteinogenic amino acid release profiles were dependent upon the disruption methods. The greater the released quantity of an amino acid during autolysis, the more the mean value fluctuated in the prepared yeast extract. Protein size fractionation of the extract using electrophoresis showed differences ranging between 1.5 and 95 kDa. All yeast extracts evidenced good nutritional potential according to FAO/WHO standards. The calculated data showed that the manufacturing method has a big impact on the proteinogenic composition of a yeast extract and the spent yeast TUM 68 used in this study can yield a protein-rich yeast extract.

This is the first report on the effect of low temperature stress applied during initial moromi fermentation on the quality and taste of soy sauce. Koji was prepared to yield initial moromi under three comparative fermentation conditions over 9 days: (1) 4 °C and 0% brine (i.e., water) (LTSF); (2) 4 °C and 16% w/w brine (LTSH); (3) 25 °C and 16% w/w brine (the control, CRTH). Greater extent of autolysis in samples was found under low temperature stress conditions (i.e., at 4 °C, a temperature much lower than the normal temperature range like 25 °C for natural microbial growth and performance). Compared to CRTH, LTSF had a two-fold increase of glutaminase activity in dregs and 65.17% increase in supernatant, and after 60 days of moromi fermentation, a 5.73% and 3.47% increase, respectively, in the contents of glutamic acid (Glu) and aspartic acid (Asp). LTSF had the highest total free amino acid content due to both the low temperature stress and absence of salt. The intensity ranking of umaminess and kokumi sensation (LTSF > LTSH > CRTH) revealed by sensory analysis followed the changing trends of their umami and sweet amino acid contents with a trend reversal in the bitter amino acid content. Low temperature (4 °C) without brine for initial moromi fermentation seemed beneficial, leading to a soy sauce product with desired taste and amino acid contents.

Metacaspases (MCs) are cysteine proteases that are implicated in programmed cell death of plants. AtMC9 (Arabidopsis thaliana Metacaspase9) is a member of the Arabidopsis MC family that controls the rapid autolysis of the xylem vessel elements, but its downstream targets in xylem remain uncharacterized. PttMC13 and PttMC14 were identified as AtMC9 homologs in hybrid aspen (Populus tremula × tremuloides). A proteomic analysis was conducted in xylem tissues of transgenic hybrid aspen trees which carried either an overexpression or an RNA interference construct for PttMC13 and PttMC14. The proteomic analysis revealed modulation of levels of both previously known targets of metacaspases, such as Tudor staphylococcal nuclease, heat shock proteins and 14-3-3 proteins, as well as novel proteins, such as homologs of the PUTATIVE ASPARTIC PROTEASE3 (PASPA3) and the cysteine protease RD21 by PttMC13 and PttMC14. We identified here the pathways and processes that are modulated by PttMC13 and PttMC14 in xylem tissues. In particular, the results indicate involvement of PttMC13 and/or PttMC14 in downstream proteolytic processes and cell death of xylem elements. This work provides a valuable reference dataset on xylem-specific metacaspase functions for future functional and biochemical analyses.

This review examines the different types of interactions between the microorganisms involved in the fermentation processes of alcoholic beverages produced all over the world from cereals or fruit juices. The alcoholic fermentation converting sugars into ethanol is usually carried out by yeasts, mainly Saccharomyces cerevisiae , which can grow directly using fruit sugars, such as those in grapes for wine or apples for cider, or on previously hydrolyzed starch of cereals, such as for beers. Some of these beverages, or the worts obtained from cereals, can be distilled to obtain spirits. Besides S. cerevisiae , all alcoholic beverages can contain other microorganisms and especially in spontaneous fermentation when starter cultures are not used. These other microbes are mostly lactic acid bacteria and other yeasts—the non- Saccharomyces yeasts. The interactions between all these microorganisms are very diverse and complex, as in any natural occurring ecosystem, including food fermentations. To describe them, we have followed a simplified ecological classification of the interactions. The negative ones are amensalism, by which a metabolic product of one species has a negative effect on others, and antagonism, by which one microbe competes directly with others. The positive interactions are commensalism, by which one species has benefits but no apparent effect on others, and synergism, by which there are benefits for all the microbes and also for the final product. The main interactions in alcoholic beverages are between S. cerevisiae and non- Saccharomyces and between yeasts and lactic acid bacteria. These interactions can be related to metabolites produced by fermentation such as ethanol, or to secondary metabolites such as proteinaceous toxins, or are feed-related, either by competition for nutrients or by benefit from released compounds during yeast autolysis. The positive or negative effects of these interactions on the organoleptic qualities of the final product are also revised. Focusing mainly on the alcoholic beverages produced by spontaneous fermentations, this paper reviews the interactions between the different yeasts and lactic acid bacteria in wine, cider, beer, and in spirits such as tequila, mezcal and cachaça.