Probiotic Potential of the Farmed Olive Flounder,Paralichthys olivaceus,Autochthonous Gut Microbiota

In recent years, considerable and growing attention has been given to the application of host-associated microorganisms as a more suitable source of probiotics in aquaculture sector. Herein, we isolated and screened the olive flounder gut microbiota for beneficial bacterial strains that might serve as potential probiotics in a low fishmeal extruded aquafeed. Among the ten identified isolates, Bacillus amyloliquefaciens SK4079 and B. subtilis SK4082 were screened out based on their heat-resistant ability as well as enzymatic and non-hemolytic activities. Although both strains were well able to utilize carboxymethyl cellulose (CMC), xylan, and soybean meal (SBM) as a single carbon source in the minimal nutrient M9 medium, B. subtilis exhibited significantly higher cellulase, xylanase, and protease activities than B. amyloliquefaciens. The two selected strains were well able to degrade the undesirable anti-nutritional component of the SBM, which would limit its utilization as protein source in aquafeed industry. Significantly higher biofilm formation capacity and notably stronger adhesive interactions with the flounder’s skin mucus were detected in B. subtilis than B. amyloliquefaciens. Immobilization of the spores from the selected strains, in a SBM complex carrier, remarkably enhances their thermal resistance at 120 °C for 5 min and different drying conditions. It was also interesting to learn that the B. subtilis spores could survive and remain viable after being sprayed onto extruded low-fish meal feed pellets for as long as 6 months. Overall, the findings of the present study could help the food/feed industries achieve their goal of developing cost-effective yet efficient products.

     

Probiotic Potential and Effects on Gut Microbiota Composition and Immunity of Indigenous Gut Lactobacilli in Apis cerana

Probiotics and Antimicrobial Proteins - This study aimed to investigate the probiotic potential of gut indigenous lactic acid bacteria (LAB) originated from Apis cerana. Six Limosilactobacillus...     

Metabolism of praziquantel in kingfish Seriola lalandi

Investigations into the metabolism of drugs used in aquatic animal therapy are useful for understanding the mechanisms of xenobiotic transformation systems and can aid the development of dosing regimens. This study investigated the metabolism of the synthetic anthelmintic praziquantel, which has application in helminthiasis treatment for several fish species including kingfish Seriola lalandi, a commercial aquaculture finfish species. At least 7 mono- or dihydroxylated derivatives of the parent compound were identified in kingfish after administration of a 150 mg kg(-1) oral praziquantel dose, paralleling findings in mammals. The structure of one representative mono-hydroxylated species that was prominent in the skin, muscle, liver, kidney and plasma of kingfish was investigated using fragmentation experiments; this revealed that hydroxylation of the parent molecule occurred in the tetrahydroisoquinoline region of praziquantel, analogous with mammalian metabolites, but different to that of the active mammalian metabolite (trans-4-OH-praziquantel). The implications of these findings with regard to biotransformation systems for this drug in mammals and fish are discussed.     

Probiotic Gut Microbiota Isolate Interacts with Dendritic Cells via Glycosylated Heterotrimeric Pili

Mapping of the microbial molecules underlying microbiota-host interactions is key to understand how microbiota preserve mucosal homeostasis. A pivotal family of such bacterial molecules are pili. Pili are proteinaceous cell wall appendages with a well-documented role in adhesion, whilst their role in immune interaction with the host is less established. Gram-positive pili are often posttranslationally modified by sortase-specific cleavage reactions and the formation of intramolecular peptide bonds. Here we report glycosylation as a new level of posttranslational modification of sortase-dependent pili of a beneficial microbiota species and its role in immune modulation. We focused on the SpaCBA pili of the model probiotic and beneficial human gut microbiota isolate Lactobacillus rhamnosus GG. A unique combination of molecular techniques, nanoscale mechanical and immunological approaches led to the identification of mannose and fucose residues on the SpaCBA pili. These glycans on the pili are recognized by human dendritic cells via the C-type lectin receptor DC-SIGN, a key carbohydrate-dependent immune tailoring pattern recognition receptor. This specific lectin-sugar interaction is moreover of functional importance and modulated the cytokine response of dendritic cells. This provides insight into the direct role bacterial glycoproteins can play in the immunomodulation of the host. Modification of the complex heterotrimeric pili of a model probiotic and microbiota isolate with mannose and fucose is of importance for the functional interaction with the host immune lectin receptor DC-SIGN on human dendritic cells. Our findings shed light on the yet underappreciated role of glycoconjugates in bacteria-host interactions.     

Probiotic Potential of Autochthonous Bacteria Isolated from the Gastrointestinal Tract of Four Freshwater Teleosts

In this study, a total of 121 bacterial strains were isolated from the gastrointestinal tract of four teleostean species, namely striped snakehead (Channa striatus), striped dwarf catfish (Mystus vittatus), orangefin labeo (Labeo calbasu) and mrigal carp (Cirrhinus mrigala), among which 8 isolates showed promising antibacterial activity against four potential fish pathogens, Aeromonas hydrophila, Aeromonas salmonicida, Aeromonas sobria and Pseudomonas fluorescens and were non-hemolytic. The isolates were further screened in response to fish bile tolerance and extracellular digestive enzyme activity. Two bacterial strains MVF1 and MVH7 showed highest tolerance and extracellular enzymes activities, and selected for further studies. Antagonistic activity of these two isolates was further confirmed by in vitro growth inhibition assay against four selected fish pathogens in liquid medium. Finally, these two bacterial strains MVF1 and MVH7 were selected as potential probiotic candidates and thus identification by partial 16S rRNA gene sequence analysis. The bacterial isolates MVF1 and MVH7 were identified as two strains of Bacillus sp.     

In Vitro Anti-staphylococcal and Anti-inflammatory Abilities of Lacticaseibacillus rhamnosus from Infant Gut Microbiota as Potential Probiotic Against Infectious Women Mastitis

Infectious mastitis is the major cause of early weaning, depriving infants of breastfeeding benefits. It is associated with an inflammatory condition of the breast and lowered resistance to infection. Drug administration during lactation often being contra-indicated, it is therefore important to consider safe therapeutic alternatives to antibiotic and anti-inflammatory therapies, such as probiotics. In this study, we investigated in vitro the probiotic potential of thirteen Lacticaseibacillus (formerly Lactobacillus) rhamnosus strains isolated from the gut microbiota of breastfed healthy infants. Strains were assessed for their β-hemolytic activity, their resistance to antibiotics, and their antimicrobial activities against strains of Staphylococcus and Streptococcus, most often involved in women mastitis. Their immunomodulating abilities were also studied using in vitro stimulation of human immune cells. None of the strains exhibited β-hemolytic activity, and all of them were sensitive to ampicillin, penicillin, tetracycline, rifampicin, erythromycin, chloramphenicol, and imipenem but showed resistance to ceftazidime, trimethoprim/sulfamethoxazole, vancomycin, and cefotaxime, reported to be chromosomally encoded and not inducible or transferable. Four L. rhamnosus strains were selected for their large anti-staphylococcal spectrum: L. rhamnosus VR1-5 and L. rhamnosus VR3-1 inhibiting S. aureus, S. epidermis, and S. warneri and L. rhamnosus CB9-2 and L. rhamnosus CB10-5 exerting antagonistic effect against S. aureus and S. epidermis strains. Antimicrobial compounds released in cell-free supernatant showed proteinaceous nature and were thermoresistant. The immune modulatory analysis of the L. rhamnosus strains revealed two strains with significant anti-inflammatory potential, highlighted by strong induction of IL-10 and a weak pro-Th1 cytokine secretion (IL-12 and IFN-γ). L. rhamnosus CB9-2 combined a large anti-staphylococcal activity spectrum and a promising anti-inflammatory profile. This strain, used individually or in a mixture, can be considered as a probiotic candidate for the management of infectious mastitis during lactation.

     

Acromyrmex Leaf-Cutting Ants Have Simple Gut Microbiota with Nitrogen-Fixing Potential

Ants and termites have independently evolved obligate fungus-farming mutualisms, but their gardening procedures are fundamentally different, as the termites predigest their plant substrate whereas the ants deposit it directly on the fungus garden. Fungus-growing termites retained diverse gut microbiota, but bacterial gut communities in fungus-growing leaf-cutting ants have not been investigated, so it is unknown whether and how they are specialized on an exclusively fungal diet. Here we characterized the gut bacterial community of Panamanian Acromyrmex species, which are dominated by only four bacterial taxa: Wolbachia, Rhizobiales, and two Entomoplasmatales taxa. We show that the Entomoplasmatales can be both intracellular and extracellular across different gut tissues, Wolbachia is mainly but not exclusively intracellular, and the Rhizobiales species is strictly extracellular and confined to the gut lumen, where it forms biofilms along the hindgut cuticle supported by an adhesive matrix of polysaccharides. Tetracycline diets eliminated the Entomoplasmatales symbionts but hardly affected Wolbachia and only moderately reduced the Rhizobiales, suggesting that the latter are protected by the biofilm matrix. We show that the Rhizobiales symbiont produces bacterial NifH proteins that have been associated with the fixation of nitrogen, suggesting that these compartmentalized hindgut symbionts alleviate nutritional constraints emanating from an exclusive fungus garden diet reared on a substrate of leaves.     

Bioavailability and pharmacokinetics of a praziquantel bolus in kingfish Seriola lalandi

Oral praziquantel (PZQ) preparations have recently been investigated for the treatment of monogeneans that infect the skin and gills of kingfish Seriola lalandi cultured in sea-cages. To evaluate an oral PZQ dosing strategy, the pharmacokinetics of a dissolved and in feed oral PZQ preparation (40 mg kg(-1) body weight) were compared with an intravenous bolus in kingfish plasma and skin using HPLC. Compared with intravenous administration, PZQ bioavailability (area under curve, AUC0-24h) was slightly improved when the drug was administered with food in both kingfish plasma (56.8% in feed vs. 50.8% in solution) and skin (55.5% in feed vs. 50.3% in solution). After oral dosing, maximum drug concentrations in skin were approximately one-third of those achieved in plasma and higher when the drug was administered in solution (5.26 microg ml(-1)) than in feed (3.96 microg ml(-1)); additionally, the time to achieve maximum PZQ concentration was similar in plasma and skin, although markedly reduced when the drug was administered in solution (1 h) than in feed (6 h). However, clearance of the drug was delayed in skin; administered as an oral formulation, PZQ concentrations in the systemic circulation fell below the limit of quantification after 24 h, but remained quantifiable (0.3 microg g(-1)) in skin at this time. These initial studies indicate that a daily treatment interval will lead to the exposure of parasites to highly variable anthelmintic concentrations, which may be sub-optimal for the treatment of monogeneans in this finfish species.     

In Vitro Assessment of Probiotic Potential of an Autochthonous Bacterial Isolate,Pseudomonas mosselii COFCAU_PMP5

Microbiology - Microbiota of the fish gut contains many beneficial bacteria that can be used as probiotics in aquaculture. In this study, several in vitro assays along with an in vivo safety...     

Evaluation of Effects of Laboratory Disinfectants on Mouse Gut Microbiota

Disturbances in the gut microbiota are known to be associated with numerous human diseases. Mice have proven to be an invaluable tool for investigating the role of the gut microbiota in disease processes. Nonexperimental factors related to maintaining mice in the laboratory environment are increasingly being shown to have inadvertent effects on the gut microbiota and may function as confounding variables. Microisolation technique is a term used to describe the common biosecurity practice of spraying gloved hands with disinfectant before handling research mice. This practice prevents contamination with pathogenic microorganisms. To investigate if exposure to disinfectants can affect the mouse gut microbiota, C57BL/6 mice were exposed daily for 27 consecutive days to commonly used laboratory disinfectants through microisolation technique. The effects of 70% ethanol and disinfectant products containing chlorine dioxide, hydrogen peroxide, or potassium peroxymonosulfate were each evaluated. Fecal pellets were collected after 7, 14, 21, and 28 d of disinfectant exposure, and cecal contents were collected at day 28. DNA extractions were performed on all cecal and fecal samples, and microbial community structure was characterized using 16S ribosomal RNA amplicon sequencing. Alpha and β diversity metrics and taxon-level analyses were used to evaluate differences in microbial communities. Disinfectant had a small but significant effect on fecal microbial communities compared with sham-exposed controls, and effects varied by disinfectant type. In general, longer exposure times resulted in greater changes in the fecal microbiota. Effects on the cecal microbiota were less pronounced and only seen with the hydrogen peroxide and potassium peroxymonosulfate disinfectants. These results indicate that laboratory disinfectant use should be considered as a potential factor that can affect the mouse gut microbiota.

The assortment of microorganisms that reside in the gastrointestinal tract of humans and animals are commonly referred to as the gut microbiota. Microbial colonization of the human gastrointestinal tract begins soon after birth.⁵⁰ The microbiota develops through interactions with host and environmental factors, reaching an equilibrium that is relatively stable and unique to each individual person.^20,58 The gut microbiota exists in a complex relationship with the health of the host. Benefits of the gut microbiota include fermentation of nondigestible dietary components into energy and nutrient sources, healthy immune system function, normal gastrointestinal motility, and anticancer effects.^21,38 Conversely, numerous diseases have been associated with variations in gut microbiota composition, which affect physiologic function and organ systems throughout the body. These range from diseases of the gastrointestinal tract itself such as with inflammatory bowel diseases to neurologic disorders such as with Parkinson disease, autism, and mood disorders.^48,56 Other conditions associated with the gut microbiota include obesity, type 2 diabetes, rheumatoid arthritis, liver diseases, lung injury, and cardiovascular diseases.^{25,37,41,44,62,69}Mice are valuable models for investigating the mechanisms involved in the associations seen between disease and gut microbial composition. Advantages of using mice for gut microbiota research include considerable scientific data available in mice, the ability to modify and select for specific genotypes, the establishment of experimental techniques that allow for precise control of the gut microbiota, and the ability to control environment factors.^35,47Nonexperimental differences in the housing environments of research mice have been shown to cause significant dissimilarity in the gut microbiota, and therefore these factors must be a critical consideration when designing experiments.³⁵ Environmental factors that have been shown to contribute to alterations in the gut microbiota include temperature, light cycle, diet, source of drinking water, method of water decontamination, bedding material, and static compared with ventilated microisolation caging.^{6,14,15,18,32,68}Most mice housed in biomedical research facilities are SPF, a term which indicates that the mice have been tested and determined to be free of an extensive list of infectious pathogens.⁵² Maintaining the SPF health status of laboratory mice through rigorous adherence to facility-defined biosecurity practices is vital for animal welfare and ensuring the validity and reproducibility of biomedical research studies.²² A standard biosecurity practice is the housing of SPF mice in sanitized or sterilized microisolation cages. When an experiment requires that mice be handled, such as for injection, gavage, or blood collection, microisolation practices are followed to preserve the aseptic cage environment. These practices include opening of cages inside of a laminar flow station that provides HEPA-filtered air and using disinfectant chemicals to spray the work surface, outer cage, and gloved hands before mice are handled.⁶⁷During microisolation technique handling of mice, some proportion of the disinfectant is inevitably transferred from the gloved hand to the fur of the mice. This disinfectant then becomes a source for oral exposure during normal mouse grooming behavior, which involves repetitive licking of the fur.⁵ Given publications indicating that oral ingestion of other antimicrobials, such as antibiotics and acidified water, result in significant changes to the mouse gut microbiota, we hypothesized that exposure to the disinfectant chemicals used in microisolation technique may affect the gut microbiota as well.^6,63 Oxidizing agents are often the disinfectant category of choice for use in this capacity as they are fast acting and broad spectrum with efficacy against the range of infectious agents of concern to SPF rodent facilities.^1,22 Ethanol, a denaturant disinfectant found ubiquitously in biomedical research laboratories, is not effective against several important nonenveloped viruses and as such is not an ideal choice for use in biosecurity practices.^22,31The primary objective of the current study was to determine if the use of disinfectant during standard microisolation handling of mice can influence the mouse gut microbiota. Differences in the gut microbial composition of mice exposed to 3 commercially available oxidizing agent formulations or a solution of 70% ethanol were evaluated by measuring community-level biodiversity metrics and taxon-level analyses. Effects on ⟨ diversity were assessed by differences in metrics including richness, which quantifies the average number of taxa represented in a microbial community, and Shannon index, which accounts for average taxonomic abundance in addition to richness.^53,55 Effects on β diversity, which describes the differences in total microbial community structure between groups, was assessed using Bray-Curtis dissimilarity.⁵³ In addition, differences in the relative abundance of individual microbial taxa between groups were assessed using the software package edgeR. A secondary objective of this study was to determine whether exposures to these disinfectants resulted in histologic lesions in the skin, gastrointestinal tract, or other tissues. The purpose of this research was to provide insight for institutions and investigators that use these disinfectants for maintaining biosecurity when handling SPF laboratory research mice.     

Robustness of Gut Microbiota of Healthy Adults in Response to Probiotic Intervention Revealed by High-Throughput Pyrosequencing

Probiotics are live microorganisms that potentially confer beneficial outcomes to host by modulating gut microbiota in the intestine. The aim of this study was to comprehensively investigate effects of probiotics on human intestinal microbiota using 454 pyrosequencing of bacterial 16S ribosomal RNA genes with an improved quantitative accuracy for evaluation of the bacterial composition. We obtained 158 faecal samples from 18 healthy adult Japanese who were subjected to intervention with 6 commercially available probiotics containing either Bifidobacterium or Lactobacillus strains. We then analysed and compared bacterial composition of the faecal samples collected before, during, and after probiotic intervention by Operational taxonomic units (OTUs) and UniFrac distances. The results showed no significant changes in the overall structure of gut microbiota in the samples with and without probiotic administration regardless of groups and types of the probiotics used. We noticed that 32 OTUs (2.7% of all analysed OTUs) assigned to the indigenous species showed a significant increase or decrease of ≥10-fold or a quantity difference in >150 reads on probiotic administration. Such OTUs were found to be individual specific and tend to be unevenly distributed in the subjects. These data, thus, suggest robustness of the gut microbiota composition in healthy adults on probiotic administration.     

Probiotic Clostridium butyricum Improves the Growth Performance,Immune Function,and Gut Microbiota of Weaning Rex Rabbits

Probiotics could promote animal growth and enhance immune function. This study investigated the effects of Clostridium butyricum (CB) on the growth performance, intestinal immune, and gut microbiota of weaning rex rabbits. A total of 60 healthy female rabbits (5-month-old) were divided equally into four groups and mated on the same day: control group (CTRL, fed with basal feed), low-dose group (LDG, fed with basal feed + 1.0 × 10³ CFU/g CB), middle-dose group (MDG, fed with basal feed + 1.0 × 10⁴ CFU/g CB), and high-dose group (HDG, fed with basal feed + 1.0 × 10⁵ CFU/g CB). Then, 30 weaning rex rabbits (35-day-old) were collected from each group for this experiment, and they were offered the same feeds as their mother. The results demonstrated that high-dose CB treatment significantly increased average daily weight gain of weaning rex rabbits. Further studies suggested that CB enhanced small intestinal digestive enzyme activity and improved mucosal morphology and antioxidant status. Supplemented with CB, small intestinal barrier function was maintained with the upregulation of mRNA levels of ZO-1, claudin, and occludin as well as the increase of sIgA production. Moreover, the relative expressions of MyD88, TLR2, and TLR4 were elevated in HDG; simultaneously, pro-inflammatory cytokines including IL-6, INF-γ, and TNF-α were decreased after CB administration. In addition, CB showed beneficial effects in improving weaning rex rabbit intestinal microflora via increasing the abundance of beneficial bacteria. Therefore, our results indicated CB can promote rex rabbit growth, which is likely to the enhancement of immune function and the improvement of intestinal microbiota.

     

Metagenomic Surveys of Gut Microbiota

Gut microbiota of higher vertebrates is host-specific. The number and diversity of the organisms residing within the gut ecosystem are defined by physiological and environmental factors,such as host genotype, habitat, and diet. Recently, culture-independent sequencing techniques have added a new dimension to the study of gut microbiota and the challenge to analyze the large volume of sequencing data is increasingly addressed by the development of novel computational tools and methods. Interestingly, gut microbiota maintains a constant relative abundance at operational taxonomic unit(OTU) levels and altered bacterial abundance has been associated with complex diseases such as symptomatic atherosclerosis, type 2 diabetes, obesity, and colorectal cancer. Therefore, the study of gut microbial population has emerged as an important field of research in order to ultimately achieve better health. In addition, there is a spontaneous, non-linear, and dynamic interaction among different bacterial species residing in the gut. Thus, predicting the influence of perturbed microbe–microbe interaction network on health can aid in developing novel therapeutics. Here, we summarize the population abundance of gut microbiota and its variation in different clinical states,computational tools available to analyze the pyrosequencing data, and gut microbe–microbe interaction networks.     

Ontogeny of body density and the swimbladder in yellowtail kingfish Seriola lalandi larvae

The ontogeny of larval body density and the morphological and histological events during swimbladder development were investigated in two cohorts of yellowtail kingfish Seriola lalandi larvae to understand the relationship between larval morphology and body density. Larvae <3 days post hatch (dph) were positively buoyant with a mean ± s.d . body density of 1·023 ± 0·001 g cm^?3. Histological evidence demonstrated that S. lalandi larvae are initially transient physostomes with the primordial swimbladder derived from the evagination of the gut ventral to the notochord and seen at 2 dph. A pneumatic duct connected the swimbladder to the oesophagus, but degenerated after 5 dph. Initial swimbladder (SB) inflation occurred on 3 dph, and the inflation window was 3–5 dph when the pneumatic duct was still connected to the gut. The swimbladder volume increased with larval age and the epithelial lining on the swimbladder became flattened squamous cells after initial inflation. Seriola lalandi developed into a physoclist with the formation of the rete mirabile and the gas‐secreting gland comprised low‐columnar epithelial cells. Larvae with successfully inflated swimbladders remained positively buoyant, whereas larvae without SB inflation became negatively buoyant and their body density gradually reached 1·030 ± 0·001 g cm^?3 by 10 dph. Diel density changes were observed after 5 dph, owing to day time deflation and night‐time inflation of the swimbladder. These results show that SB inflation has a direct effect on body density in larval S. lalandi and environmental factors should be further investigated to enhance the rate of SB inflation to prevent the sinking death syndrome in the early life stage of the fish larvae.     

Probiotic Lactobacillus johnsonii BS15 Promotes Growth Performance,Intestinal Immunity,and Gut Microbiota in Piglets

Probiotics and Antimicrobial Proteins - Numerous studies have investigated the beneficial effects of Lactobacillus johnsonii strain BS15 on mice and broilers. This study aimed to understand the...     

Compound Probiotic Ameliorates Acute Alcoholic Liver Disease in Mice by Modulating Gut Microbiota and Maintaining Intestinal Barrier

Probiotics and Antimicrobial Proteins - Alcoholic liver disease (ALD) is a worldwide health threaten lack of effective treatment. Gut dysbiosis and concomitant augmented intestinal permeability are...     

Prebiotic Potential of a Maize-Based Soluble Fibre and Impact of Dose on the Human Gut Microbiota

Dietary management of the human gut microbiota towards a more beneficial composition is one approach that may improve host health. To date, a large number of human intervention studies have demonstrated that dietary consumption of certain food products can result in significant changes in the composition of the gut microbiota i.e. the prebiotic concept. Thus the prebiotic effect is now established as a dietary approach to increase beneficial gut bacteria and it has been associated with modulation of health biomarkers and modulation of the immune system. Promitor™ Soluble Corn Fibre (SCF) is a well-known maize-derived source of dietary fibre with potential selective fermentation properties. Our aim was to determine the optimum prebiotic dose of tolerance, desired changes to microbiota and fermentation of SCF in healthy adult subjects. A double-blind, randomised, parallel study was completed where volunteers (n = 8/treatment group) consumed 8, 14 or 21 g from SCF (6, 12 and 18 g/fibre delivered respectively) over 14-d. Over the range of doses studied, SCF was well tolerated Numbers of bifidobacteria were significantly higher for the 6 g/fibre/day compared to 12g and 18g/fibre delivered/day (mean 9.25 and 9.73 Log10 cells/g fresh faeces in the pre-treatment and treatment periods respectively). Such a numerical change of 0.5 Log10 bifidobacteria/g fresh faeces is consistent with those changes observed for inulin-type fructans, which are recognised prebiotics. A possible prebiotic effect of SCF was therefore demonstrated by its stimulation of bifidobacteria numbers in the overall gut microbiota during a short-term intervention.