Characterization and Biological Activity of a Novel Exopolysaccharide Produced by Pediococcus pentosaceus SSC–12 from Silage

In this study, 22 strains of exopolysaccharides-producing lactic acid bacteria were isolated from silage, and the strain SSC–12 with the highest exopolysaccharide (EPS) production was used as the test strain. The SSC–12 was identified as Pediococcus pentosaceus, based upon 16S rDNA gene sequencing and Neighbor Joining (NJ) phylogenetic analysis. The analysis of the kinetic results of EPS generation of SSC–12 showed that the EPS generation reached the maximum value at 20 h of culture. The characterization study showed the EPS produced by SSC–12 was a homogeneous heteropolysaccharide comprising glucose (42.6%), mannose (28.9%), galactose (16.2%), arabinose (9.4%), and rhamnose (2.9%). The EPS had good antioxidant activity, especially the activity of scavenging hydroxyl free radicals. At the same time, the EPS also had strong antibacterial ability and could completely inhibit the growth of Staphylococcus aureus. The EPS produced by the Pediococcus pentosaceus SSC–12 can be used as a biologically active product with potential application prospects in the feed, food, and pharmaceutical industries.

Selection of Autochthonous LAB Strains of Unripe Green Tomato towards the Production of Highly Nutritious Lacto-Fermented Ingredients

Lactic fermentation of unripe green tomatoes as a tool to produce food ingredients is a viable alternative for adding value to industrial tomatoes unsuitable for processing and left in large quantities in the fields. Fermentation using starter cultures isolated from the fruit (plant-matrix adapted) can have advantages over allochthonous strains in obtaining fermented products with sensory acceptability and potentially probiotic characteristics. This paper details the characterisation of the unripe green tomato lactic microbiota to screen LAB strains for use as starter cultures in fermentation processes, along with LAB strains available from INIAV’s collection. Morphological, biochemical (API system), and genomic (16S rDNA gene sequencing) identification showed that the dominant LAB genera in unripe green tomato are Lactiplantibacillus, Leuconostoc, and Weissella. Among nine tested strains, autochthonous Lactiplantibacillus plantarum and allochthonous Weissella paramesenteroides showed tolerance to added solanine (200 ppm) and the best in vitro probiotic potential. The results indicate that the two LAB strains are promising candidates for manufacturing probiotic fermented foods from unripe green tomatoes.

Antioxidant Potential of Pediococcus Pentosaceus Strains from the Sow Milk Bacterial Collection in Weaned Piglets

Background: In modern animal husbandry, breeders pay increasing attention to improving sow nutrition during pregnancy and lactation to favor the health of neonates. Sow milk is a main food for piglets during their first three weeks of life, which is not only a rich repository of essential nutrients and a broad range of bioactive compounds, but also an indispensable source of commensal bacteria. Maternal milk microorganisms are important sources of commensal bacteria for neonatal gut. Bacteria from maternal milk may serve as an additive to confer a health benefit on the composition of the indigenous microbiota of piglets. Methods: The sow milk microbiota was collected using the culturomics methods of Continuous Culture and Interval Sampling, following by the identification of 16S rDNA gene sequences. To screen potential probiotics, the functional evaluation was conducted to assess their antagonistic activity against pathogens in vitro and evaluate their resistance against oxidative stress in damaged Drosophila induced by paraquat. In piglet feeding trial, a total of 54 newborn suckling piglets were chosen from nine sows and randomly assigned to three treatments with different concentration of a candidate strain. Multiple approaches were carried out to verify its antioxidant function including western blotting, enzyme activity analysis, metabolomics and 16S rDNA sequencing. Results: The 1240 isolates were screened out from the sow milk microbiota and grouped into 271 bacterial taxa based on a nonredundant set of 16S rDNA gene sequences. Among 80 Pediococcus isolates, a new Pediococcus pentosaceus strain (SMM914) showed best performance in inhibition ability against swine pathogens and in Drosophila model challenged by paraquat. Pretreatment of piglets with SMM914 induces the Nrf2-Keap1 antioxidant signaling pathway and greatly affected the pathways of amino acid metabolism and lipid metabolism in plasma. In colon, Lactobacillus was significantly increased in the high dose of SMM914 group compared with the control group. Conclusion: SMM914 functions as a promising probiotic conferring antioxidant capacity by activating the Nrf2-Keap1 antioxidant signaling pathway in piglets. Our study provided useful resources for deeply understanding the relationships between the maternal microbiota and offspring.

Bio-fertilizers’ protocol for controlling root knot nematode Meloidogyne javanica infecting peanut fields

Background Plant parasitic nematodes create serious threat to crop production. In Egypt root knot nematode, Meloidogyne spp. has been considered to be a limiting factor in the production of most crops of which the Peanut (Arachis hypogaea L.) is an important legume and oil crop. Therefore, management of root knot nematodes Meloidogyne spp. is an obligatory challenge. Microbial organisms are extensively used as eco-friendly tools for controlling plant parasitic nematodes as alternative to chemical nematicides. The effectiveness of the commercial bacterial bio-fertilizers NPK containing Bacillus polymyxa, B. circulance, B. megaterium, Pseudomans spp.; the nitrogen fixative bacteria Azotobacter chroocoocum and the bacterial isolate NRC211 were evaluated against the root knot nematode, Meloidogyne javanica infecting peanut plants under field conditions. Identification of the bacterial isolate was made through PCR amplification and sequencing of 16S rDNA gene. Results Sequencing of 16S rDNA gene revealed that the bacterial isolate NRC211 had 100% similarity with Bacillus wiedmannii strain FSL W8-0169 16S ribosomal RNA. This Bacillus was recorded for the first time under accession number LC626774 on GenBank data base as B. wiedmannii NRC211. Recorded data revealed that all the tested treatments whether single or combined in soil naturally infested with M. javanica, resulted in variable significant reduction in the nematode reproductive parameters with a considerable increase in crop production and oil content of peanut plant. These results were improved by increasing the frequency of application of the bio-agents. In this respect the repeated combined treatment of A. chroococcum and B. wiedmannii NRC211 treatment overwhelmed all other treatments in decreasing nematode reproductive parameters with percentage reductions of 94.8, 79.0 and 80.1% in M. javanica juveniles in soil, galls and egg masses, respectively. This was associated with slight increase in peanut oil content than the untreated control. The repeated combined treatment of NPK plus A. chroococcum produced the highest increase 608.7%, and 72.7% in crop production and plant growth parameters, respectively than the control. While, the oil content in this treatment was increased up to 47.4 g/kg. Conclusion It was concluded that B. wiedmanni NRC211 is an eco-friendly bio agent that can be applied with other commercial microbial bio-fertilizers in bio-integrating programs for controlling M. javanica infecting peanut plants.

The effect of sodium carboxymethyl starch with high degree of substitution on defecation

Sodium carboxymethyl starch (CMS-Na), a kind of food additive with high degree of substitution, is also known as a prebiotic. The aim of this study was to determine the effect of CMS-Na on defecation. Constipated mouse model was prepared by loperamide. Normal rats were also used in the study. Short-chain fatty acids in rat feces were detected by gas chromatography. The bacterial communities in rat feces were identified by 16S rDNA gene sequencing. 5-hydroxytryptamine (5-HT) and tryptophan hydroxylase 1 (Tph1) were measured by ELISA. The results showed that CMS-Na increased the fecal granule counts and intestinal propulsion rate in constipated mice. The contents of water, acetic acid, propionic acid and n-butyrate in feces, Tph1 in colon and 5-HT in serum of rats were increased. In addition, CMS-Na shortened the colonic transport time in rats. The 16S rDNA gene sequencing results indicated that CMS-Na increased the relative abundance of Alloprevotella and decreased the proportion of Lactobacillus. However, the biodiversity of the normal intestinal flora was not altered. In conclusion, CMS-Na can promote defecation in constipated mice. The mechanism may be related to the regulation of Alloprevotella and Lactobacillus in colon, the increase of short-chain fatty acids, and the promotion of the synthesis of Tph1 and 5-HT.

Isolation and Toxigenic Characteristics of a Tetrodotoxin Producing Bacteria

Tetrodotoxin (TTX) is a kind of neurotoxin with great scientific research and medicinal value. Studies have confirmed that microorganism is one of the important sources of TTX. However, the specific biosynthetic and regulatory pathway of TTX production by microorganisms are still unclear. In this paper, out of twelve symbiotic bacteria screened from the wild puffer fish (Fugu rubripes) collected from coastal sea area of Ningbo (Zhejiang, China), a strain named He-1 isolated from the liver of puffer fish was found can produce TTX. It was identified as Bacillus cereus based on physiological, biochemical characteristics and 16S rDNA gene analysis. However, the strain He-2, which is the subculture strain of He-1, cannot produce TTX under the normal culture condition. Further study showed that by adding the metabolite of the He-1, the ability of TTX producing of He-2 could be restored. A peptide compound was obtained by separating and purifying of the metabolites from strain He-1, which could induce the TTX production of strain He-2. This work will provide a new way for further revealing the anabolic pathway of TTX.

Isolation and Characterization of Biosurfactant-Producing Bacteria from Amapaense Amazon Soils

The objective of this research was to perform screening of biosurfactant-producing bacteria from Amapaense Amazon soils. Floodplain- and upland-forest soils of three municipalities of the Amapá state were isolated and identified. The isolates were cultured in nutrient broth with olive oil, and their extracts were evaluated according to drop collapse, oil dispersion, emulsification, and surface tension tests. From three hundred and eighteen isolates, the 43 bacteria were selected and identified by 16S rDNA gene sequencing, indicating the presence of three different genera, Serratia, Paenibacillus, and Citrobacter. The extracellular biosurfactant production pointed out the 15 most efficient bacteria that presented high emulsification capacity (E24 > 48%) and stability (less than 10% of drop after 72 h) and great potential to reduce the surface tension (varying from 49.40 to 34.50 mN·m−1). Cluster analysis classified genetically related isolates in different groups, which can be connected to differences in the amount or the sort of biosurfactants. Isolates from Serratia genus presented better emulsification capacity and produced a more significant surface tension drop, indicating a promising potential for biotechnological applications.

Descriptive Study of Gut Microbiota in Infected and Colonized Subjects by Clostridiodes difficile

Clostridiodes difficile can lead to a range of situations from the absence of symptoms (colonization) to severe diarrhea (infection). Disruption of gut microbiota provides an ideal environment for infection to occur. Comparison of gut microbiota of infected and colonized subjects could provide relevant information on susceptible groups or protectors to the development of infection, since the presence of certain genera could be related to the inhibition of transition from a state of colonization to infection. Through high-throughput sequencing of 16S rDNA gene, we performed alpha and beta diversity and composition studies on 15 infected patients (Group CDI), 15 colonized subjects (Group P), and 15 healthy controls (Group CTLR). A loss of alpha diversity and richness and a different structure have been evidenced in the CDI and P groups with respect to the CTRL group, but without significant differences between the first two. In CDI and P groups, there was a strong decrease in phylum Firmicutes and an expansion of potential pathogens. Likewise, there was a loss of inhibitory genus of C. difficile germination in infected patients that were partially conserved in colonized subjects. Therefore, infected and colonized subjects presented a gut microbiota that was completely different from that of healthy controls, although similar to each other. It is in composition where we found that colonized subjects, especially in minority genera, presented differences with respect to those infected.

Molecular genetics and phylogeny of Ephippigera species group of genus Saga Charpentier, 1825 (Orthoptera: Ensifera: Saginae) in Anatolia

The genus Saga, one of the largest predatory bush crickets and insects in the Palearctic region, is represented by nine species in Anatolia. According to the morphological and bioacoustics evaluations of the Ephippigera species group belonging to the Saga genus, it consists of three species: S. ephippigera, S. syriaca, and S. hakkarica. To the present, however, there is no molecular genetic research on this species group. In our study, by using the 16S rDNA gene sequences, the molecular phylogeny of this species group is revealed and the morphological species hypotheses has been tested. In the study, specimens belonging to S. ephippigera, S. syriaca, and S. hakkarica were collected between the years of 2018 and 2020 with sweep netting; the DNA isolation, polymerase chain reactions, purifications and double-stranded sequencing were performed to obtain 16S rDNA gene sequences. Sequences were evaluated phylogenetically by performing alignment, phylogenetic analysis (Maximum Likelihood and Bayesian Inference), and dating of cladogenetic events, respectively. The monophyletic and species hypotheses of all three species have been supported by molecular data. It has been determined that the S. ephippigera species is the origin lineages of the Ephippigera species group which diverged approximately 0.4 million years ago; it has been also revealed that the climatic changes in the glacial and inter-glacial periods in the Middle Pleistocene period affected greatly the evolution of the species group.