Microbiota Of Fish Research Articles

High-throughput sequencing of gut microbiota in rainbow trout (Oncorhynchus mykiss) fed larval and pre-pupae stages of black soldier fly (Hermetia illucens)

Black soldier fly (Hermetia illucens) meal is a potential alternative to fishmeal and plant proteins in diets for farmed fish since it can be produced on organic waste substrates, requires little energy and water inputs and contains high levels of essential amino acids. Recent studies have partially replaced fishmeal with black soldier fly meal, however, research on their impact on gut microbiota of fish is limited. In a five week experiment, juvenile rainbow trout (Oncorhynchus mykiss) were fed either a reference diet based on fishmeal or three diets with 30% inclusion of black soldier fly meals in the form of pre-pupae, larvae or defatted-larvae. The combined luminal content and mucosa were collected from the distal intestine of three fish per tank with four tanks per diet (n = 12) and 16S rRNA gene amplicons were sequenced using the Illumina MiSeq platform. Feeding the insect-based diets increased the alpha-diversity of bacteria and abundance of lactic acid bacteria, which may be due to the addition of dietary chitin. Compared with fishmeal, feeding insects resulted in higher abundance of phyla Firmicutes and Actinobacteria with lower abundance of Proteobacteria. Fish fed the full-fat meals had higher abundance of Corynebacterium that was attributed to its ability to produce lipase and the high content of dietary lipids as a substrate. Bacillaceae was increased in fish fed both larvae diets and unchanged in the pre-pupae diet, which indicated that life-cycle stage of the insect influenced the gut microbiota. Based on these results, we found that feeding black soldier flies increased diversity and altered the composition of gut bacteria of rainbow trout, which were further influenced by life-cycle stage and lipid content of the insect meal.

Genes de virulencia en Aeromonas spp. (Aeromonadales: Aeromonadaceae) aisladas de Oreochromis spp. (Perciformes: Cichlidae) para consumo humano en México

The genus Aeromonas are widely distributed in aquatic ecosystems are Gram-negative rods, oxidase-positive, and glucose-fermenting, considered emerging pathogens in humans. Aeromonas belongs to the fish microbiota, these microorganisms have a diversity of virulence factors responsible for a variety of infections in humans mainly gastrointestinal diseases. The presence of Aeromonas in products intended for consumption with high commercial demand such as tilapia generates sanitary concern due to the pathogenic potential of this bacteria. In this context, identification of virulence genes in strains of Aeromonas isolated in Oreochromis spp. intended for human consumption in Reynosa, Tamaulipas, Mexico is important due to the lack of molecular studies in this geographical area. In the present study the pathogenic potential of 15 strains of Aeromonas (A. veronii, A. hydrophila and A. schubertii) from Oreochromis spp. for human consumption were analyzed. Through PCR six virulence genes were analyzed (alt, ast, aerA, hlyA, gcat and stx1) and the strains used as control were: Aeromonas hydrophila subsp. hydrophila ATCC 7966, Aeromonas caviae 429865 INP, Escherichia coli O157: H7 and Escherichia coli K12. El 100 % (n = 15) of the strains harbored at least one virulence gene, aerA gene was detected in 86.66% of the analyzed strains, while ast and stx1 genes were not identified. Moreover, Aeromonas strains had associated genes in the same strain: aerA / gcat, alt / aerA, alt / aerA / gcat / hlyA and alt / aerA / gcat, of which aerA / gcat were observed mostly in A. veronii, while A. hydrophila had the highest associations. These findings indicate that the strains of Aeromonas isolated in Oreochromis spp. have the potential to cause human diseases, and therefore, this species used as food, could be a vehicle for infections caused by Aeromonas. It also allows to provide information on this emerging microorganism to effectively treat and control any epidemiological event caused by Aeromonas spp. in the future.

Transcriptome analysis of grass carp provides insights into disease-related genes and novel regulation pattern of bile acid feedback in response to lithocholic acid

Gut microbiota and its metabolites affect lipid metabolism and liver health. Grass carp (Ctenopharyngodon idellus) often suffers from functional disorders of liver and gallbladder, which is usually accompanied by accumulation of lipids in the liver. Here we studied liver transcriptome and microbial fluctuation in the gut in response to dietary lithocholic acid (LCA; a metabolite of gut microbiota) supplementation in grass carp. A total of 1802 differentially expressed genes (DEGs) were identified between LCA and Control groups according to the liver transcriptome analysis. Of these DEGs, 8 nonalcoholic fatty liver disease-related genes were all upregulated, and 11 of 12 cancer related genes were upregulated in the LCA group. We also found significantly upregulated bile acid receptor, hepatocyte nuclear factor 4-beta (HNF4B), in the LCA group. The red oil O staining of liver showed a higher abundance of lipid droplets in the LCA group. LCA also changed the composition of gut microbiota, with increasing proportion of Proteobacteria and Firmicutes, whereas Fusobacteria decreased. Co-occurrence between DEGs and microbial taxa was mainly identified between signal transduction and genetic information processing-related genes and Proteobacteria and Firmicutes taxa. Overall, our results elucidate the effects of LCA on liver and gut microbiota in fish, and suggest a novel pattern of bile acid feedback in grass carp. These results contribute to our understanding of the interplay between gut microbiota and liver diseases in fish, and offer insights into putative treatments of liver diseases in grass carp.

How Can We Define “Optimal Microbiota?”: A Comparative Review of Structure and Functions of Microbiota of Animals, Fish, and Plants in Agriculture

All multicellular organisms benefit from their own microbiota, which play important roles in maintaining the host nutritional health and immunity. Recently, the number of studies on the microbiota of animals, fish, and plants of economic importance is rapidly expanding and there are increasing expectations that productivity and sustainability in agricultural management can be improved by microbiota manipulation. However, optimizing microbiota is still a challenging task because of the lack of knowledge on the dominant microorganisms or significant variations between microbiota, reflecting sampling biases, different agricultural management as well as breeding backgrounds. To offer a more generalized view on microbiota in agriculture, which can be used for defining criteria of “optimal microbiota” as the goal of manipulation, we summarize here current knowledge on microbiota on animals, fish, and plants with emphasis on bacterial community structure and metabolic functions, and how microbiota can be affected by domestication, conventional agricultural practices, and use of antimicrobial agents. Finally, we discuss future tasks for defining “optimal microbiota,” which can improve host growth, nutrition, and immunity and reduce the use of antimicrobial agents in agriculture.

Shifts in the Skin-Associated Microbiota of Hatchery-Reared Common Snook Centropomus undecimalis During Acclimation to the Wild

The skin-associated microbiota of fish competes against pathogens for space and nutrients, preventing colonization by harmful bacteria encountered during environmental transitions such as those faced during stock enhancement. Thus, alterations in bacterial community structure during release of cultured fish have important implications for health of these individuals. This study investigated microbiota structure during acclimation of juvenile hatchery-reared common snook Centropomus undecimalis to the wild by comparing skin-associated microflora among snook in captivity, after 48 h of acclimation at release sites, and from the wild. After two days of acclimation, the microbiota of hatchery-reared snook mirrored that observed on wild snook. Relative abundances of potential pathogens were higher in captive fish, whereas acclimated and wild fish harbored bacterial taxa influenced by geographical factors and water quality at release sites. Predicted microbiota function of acclimated and wild fish showed higher production of protective amino acids and antimicrobials, identifying a mechanism for microbial supplementation of the immune defense of these fish. The two-day transition to wild-type microbiota suggests a temporal scale of hours associated with bacterial succession indicating that the microbiota, whose structure is vital to fish health, aids in acclimation of fish to new environments during stock enhancement efforts.

Composition of Intestinal Microbiota in Two Lines of Rainbow Trout (Oncorhynchus Mykiss) Divergently Selected for Muscle Fat Content.

Background:Recently, studies suggest that gut microbiota contributes to the development of obesity in mammals. In rainbow trout, little is known about the role of intestinal microbiota in host physiology.Objective:The aim of this study was to investigate the link between intestinal microbiota and adiposity, by high-throughput 16S RNA gene based illumina Miseq sequencing in two rainbow trout lines divergently selected for muscle lipid content. Fish from these two lines of rainbow trout are known to have a differing lipid metabolism.Methods:Samples from the two lines (L for lean and F for fat) were collected from Midgut (M) and Hindgut (H) in juvenile fish (18 months) to compare intestinal microbiota diversity.Results:Whatever the lines and intestinal localisation, Proteobacteria, Firmicutes and Actinobacteria are the dominant phyla in the bacterial community of rainbow trout (at least 97%). The results indicate that richness and diversity indexes as well as bacterial composition are comparable between all groups even though 6 specific OTUs were identified in the intestinal microbiota of fish from the fat line and 2 OTUs were specific to the microbiota of fish from the lean line. Our work contributes to a better understanding in microbial diversity in intestinal microbiota of rainbow trout.Conclusion:Altogether, our study indicates that no major modification of the intestinal microbiota is induced by selection for muscle lipid content and associated metabolic changes. Finally, we identified members of core microbiota in rainbow trout.

Therapeutic Effect of Intestinal Autochthonous Lactobacillus reuteri P16 Against Waterborne Lead Toxicity in Cyprinus carpio.

Harmful effects of heavy metals are myriad. Lead (Pb) from soil and atmosphere contaminates water bodies and affects the aquatic animals. Our previous study confirmed the in vitro probiotic potential of Lactobacillus reuteri against Pb toxicity, but further investigation is necessary for gaining insights into the related protection mode. Therefore, in this study, we investigated the protective effects of the potential probiotic L. reuteri P16 against waterborne Pb exposure-induced toxicity in the freshwater fish Cyprinus carpio. Fish (average weight: 23.16 ± 0.73 g) were allocated to four groups (control, Pb only, Pb + L. reuteri P16, and L. reuteri P16 only) and Pb groups were exposed to waterborne Pb (1 mg L−1) for 6 weeks. L. reuteri P16 (108 CFU g−1) supplemented diet was provided twice daily. Growth performances, hemato-biochemical parameters, innate immune responses, intestinal microbiota, and Pb accumulation in tissues were measured at the end of the trial. When the fish were exposed to Pb, dietary supplementation of L. reuteri P16 effectively decreased mortality and accumulation of Pb in tissues, and improved the growth performance. Co-treatment with Pb and L. reuteri P16 alleviated Pb exposure-induced oxidative stress, reversed alterations in hemato-biochemical parameters, improved innate immune parameters, and restored intestinal enzymatic activities. Moreover, L. reuteri P16 supplementation reversed the changes in intestinal microbiota in Pb-exposed fish. Furthermore, Pb exposure decreased the expressions of pro-inflammatory cytokines (TNF-α, IL-1β). However, the expression of heat shock proteins (HSP70 and HSP90) increased, which might have increased the cellular stress. Interestingly, the Pb-induced alterations of gene expressions were reversed by L. reuteri P16 supplementation. Thus, dietary administration of the potential probiotic L. reuteri P16 had several beneficial effects on growth performance and immune responses, decreased Pb accumulation in tissues, and reversed alterations in hematological responses of C. carpio. Furthermore, it offered direct protection against Pb-induced oxidative stress. Therefore, L. reuteri P16 may be a novel dietary supplement for enhancing growth performance and preventing Pb-exposure-induced toxicity in fish in aquaculture and aquatic products.

The effect of fishmeal inclusion and prebiotic supplementation on the hindgut faecal microbiota of farmed Tasmanian Atlantic salmon (Salmo salar L.).

Factors such as seasonal temperature and diet components, for example, fishmeal (FM) inclusion, can influence the composition of the gut microbiota of fish. In this study, we examined changes in the gut bacterial populations, in particular lactic acid bacteria (LAB), of farmed Tasmanian Atlantic salmon in response to different diets, during periods of higher water temperature. Between December 2011 and March 2012 hindgut faecal samples were collected from Atlantic salmon from a commercial fish farm in south of Hobart, Tasmania, fed with one of four trial diets containing either high or low FM inclusion levels with or without prebiotics. Overall there was little difference in the cultivatable bacterial populations in response to varying levels of FM and prebiotic supplementation, with LAB counts decreasing in response to increased water temperatures. However, it was observed that the high FM diet supported the presence of LAB in January, when these were not detected in the low FM diets. Our study indicates that the inclusion of higher amounts of FM rather than the addition of prebiotics has a greater effect on LAB colonization of the gut in Atlantic salmon. This study highlights the importance of the new fish feeds for promoting salmon health in aquaculture industry.

Intestinal microbiota composition is altered according to nutritional biorhythms in the leopard coral grouper (Plectropomus leopardus).

Aquaculture is currently a major source of fish and has the potential to become a major source of protein in the future. These demands require efficient aquaculture. The intestinal microbiota plays an integral role that benefits the host, providing nutrition and modulating the immune system. Although our understanding of microbiota in fish gut has increased, comprehensive studies examining fish microbiota and host metabolism remain limited. Here, we investigated the microbiota and host metabolism in the coral leopard grouper, which is traded in Asian markets as a superior fish and has begun to be produced via aquaculture. We initially examined the structural changes of the gut microbiota using next-generation sequencing and found that the composition of microbiota changed between fasting and feeding conditions. The dominant phyla were Proteobacteria in fasting and Firmicutes in feeding; interchanging the dominant bacteria required 12 hours. Moreover, microbiota diversity was higher under feeding conditions than under fasting conditions. Multivariate analysis revealed that Proteobacteria are the key bacteria in fasting and Firmicutes and Fusobacteria are the key bacteria in feeding. Subsequently, we estimated microbiota functional capacity. Microbiota functional structure was relatively stable throughout the experiment; however, individual function activity changed according to feeding conditions. Taken together, these findings indicate that the gut microbiota could be a key factor to understanding fish feeding conditions and play a role in interactions with host metabolism. In addition, the composition of microbiota in ambient seawater directly affects the fish; therefore, it is important to monitor the microbiota in rearing tanks and seawater circulating systems.

Comparison of Channel Catfish and Blue Catfish Gut Microbiota Assemblages Shows Minimal Effects of Host Genetics on Microbial Structure and Inferred Function.

The microbiota of teleost fish has gained a great deal of research attention within the past decade, with experiments suggesting that both host-genetics and environment are strong ecological forces shaping the bacterial assemblages of fish microbiomes. Despite representing great commercial and scientific importance, the catfish within the family Ictaluridae, specifically the blue and channel catfish, have received very little research attention directed toward their gut-associated microbiota using 16S rRNA gene sequencing. Within this study we utilize multiple genetically distinct strains of blue and channel catfish, verified via microsatellite genotyping, to further quantify the role of host-genetics in shaping the bacterial communities in the fish gut, while maintaining environmental and husbandry parameters constant. Comparisons of the gut microbiota among the two catfish species showed no differences in bacterial species richness (observed and Chao1) or overall composition (weighted and unweighted UniFrac) and UniFrac distances showed no correlation with host genetic distances (Rst) according to Mantel tests. The microbiota of environmental samples (diet and water) were found to be significantly more diverse than that of the catfish gut associated samples, suggesting that factors within the host were further regulating the bacterial communities, despite the lack of a clear connection between microbiota composition and host genotype. The catfish gut communities were dominated by the phyla Fusobacteria, Proteobacteria, and Firmicutes; however, differential abundance analysis between the two catfish species using analysis of composition of microbiomes detected two differential genera, Cetobacterium and Clostridium XI. The metagenomic pathway features inferred from our dataset suggests the catfish gut bacterial communities possess pathways beneficial to their host such as those involved in nutrient metabolism and antimicrobial biosynthesis, while also containing pathways involved in virulence factors of pathogens. Testing of the inferred KEGG (Kyoto Encyclopedia of Genes and Genomes) pathways by DESeq2 revealed minor difference in microbiota function, with only two metagenomic pathways detected as differentially abundant between the two catfish species. As the first study to characterize the gut microbiota of blue catfish, our study results have direct implications on future ictalurid catfish research. Additionally, our insight into the intrinsic factors driving microbiota structure has basic implications for the future study of fish gut microbiota.

The Gut Microbiota of Marine Fish.

The body of work relating to the gut microbiota of fish is dwarfed by that on humans and mammals. However, it is a field that has had historical interest and has grown significantly along with the expansion of the aquaculture industry and developments in microbiome research. Research is now moving quickly in this field. Much recent focus has been on nutritional manipulation and modification of the gut microbiota to meet the needs of fish farming, while trying to maintain host health and welfare. However, the diversity amongst fish means that baseline data from wild fish and a clear understanding of the role that specific gut microbiota play is still lacking. We review here the factors shaping marine fish gut microbiota and highlight gaps in the research.

A meta-barcoding approach to assess and compare the storage temperature-dependent bacterial diversity of gilt-head sea bream (Sparus aurata) originating from fish farms from two geographically distinct areas of Greece

Bacterial diversity of whole gilt-head sea bream (Sparus aurata L. 1758) originating from Ionian and Aegean Sea aquaculture farms and stored at 0 (ice), 4 and 8 °C was determined by 16S rRNA gene amplicon sequencing method using the Illumina's MiSeq platform. The composition of Aerobic Plate Counts (APC) was also monitored by 16S rRNA gene sequencing. The rejection time point of sea bream from either area, as determined by sensory evaluation, was about 14, 6 and 3 days at 0, 4 and 8 °C, respectively. APC was approximately 4.5 log cfu/g at day 0 and ranged from 7.5 to 8.5 log cfu/g at sensory rejection. Culture-depended analysis showed that Pseudomonas and Shewanella were the most abundant microorganisms grown on plates for both seas. Moreover, culture-independent analysis of DNA extracted directly from fish flesh showed that sea bream originating from different geographical areas exhibited different bacterial diversity. Pseudomonas and Psychrobacter were the dominant microorganisms of chill-stored fish from Ionian (apart from 8 °C, where Carnobacterium dominated) and Aegean Sea, respectively. In addition, small changes of storage temperature greatly affected bacterial microbiota of stored fish. Various bacterial species, not detected by conventional microbiological methods, were also revealed through 16S amplicon sequencing. In conclusion, the use of NGS approach is a promising methodology for assessing bacterial diversity of sea bream originating from different geographical areas and stored at various temperatures.

The Vibrio cholerae type VI secretion system can modulate host intestinal mechanics to displace gut bacterial symbionts

Host-associated microbiota help defend against bacterial pathogens; however, the mechanisms by which pathogens overcome this defense remain largely unknown. We developed a zebrafish model and used live imaging to directly study how the human pathogen Vibrio cholerae invades the intestine. The gut microbiota of fish monocolonized by symbiotic strain Aeromonas veronii was displaced by V. cholerae expressing its type VI secretion system (T6SS), a syringe-like apparatus that deploys effector proteins into target cells. Surprisingly, displacement was independent of T6SS-mediated killing of A. veronii, driven instead by T6SS-induced enhancement of zebrafish intestinal movements that led to expulsion of the resident microbiota by the host. Deleting an actin cross-linking domain from the T6SS apparatus returned intestinal motility to normal and thwarted expulsion, without weakening V. cholerae's ability to kill A. veronii in vitro. Our finding that bacteria can manipulate host physiology to influence intermicrobial competition has implications for both pathogenesis and microbiome engineering.

Composition and antimicrobial resistance profile of Gram‐negative microbiota prevalent in aquacultured fish

AbstractAquatic environments are one of the most favorable settings for acquisition and dissemination of antimicrobial resistance. The aims of this study were twofold: (1) to isolate and identify antimicrobial resistant Gram‐negative bacteria that are prevalent in fish kept in open aquaculture ponds and (2) to analyse their antimicrobial resistance profiles. One hundred and nineteen isolates from fish resistant to at least one antimicrobial were investigated. The most predominant genera were Pseudomonas, Chryseobacterium, Enterobacteriaceae, and Aeromonas. The highest numbers of multiresistant isolates were found within Pseudomonas spp., and the lowest, in Aeromonas spp. Resistance to beta‐lactams was the most common among all of the bacterial genera. The absence or very low rates of resistance to fluoroquinolones, the small number of clinically important resistance genes, and bacterial taxonomic composition suggest that the most widespread resistant bacteria in aquacultured fish are autochthonous microbiota.Practical applicationsThe majority of antimicrobial resistant bacteria that are prevalent in aquacultured fish from open ponds can be treated as natural (autochthonous microbiota). In this study, only a minority of resistant bacteria detected in fish could be associated with polluted waters originating from animal farms or human healthcare facilities. Some strains, particularly the family Enterobacteriaceae that is found in fish, contain clinically important resistance determinants that potentially can be transmitted to human microbiota in cases of inappropriate processing of fish intended for food. With regard to antimicrobial resistant bacteria, measures for control of inflowing waters to aquaculture ponds should be taken in addition to monitoring fish microbiota and its’ resistance should be investigated on a regular basis.

The Inner Workings of the Outer Surface: Skin and Gill Microbiota as Indicators of Changing Gut Health in Yellowtail Kingfish.

The mucosal surfaces and associated microbiota of fish are an important primary barrier and provide the first line of defense against potential pathogens. An understanding of the skin and gill microbial assemblages and the factors which drive their composition may provide useful insights into the broad dynamics of fish host–microbial relationships, and may reveal underlying changes in health status. This is particularly pertinent to cultivated systems whereby various stressors may led to conditions (like enteritis) which impinge on productivity. As an economically important species, we assessed whether the outer-surface bacterial communities reflect a change in gut health status of cultivated Yellowtail Kingfish (Seriola lalandi). Active bacterial assemblages were surveyed from RNA extracts from swabs of the skin and gills by constructing Illumina 16S rRNA gene amplicon libraries. Proteobacteria and Bacteroidetes were predominant in both the skin and gills, with enrichment of key β-proteobacteria in the gills (Nitrosomonadales and Ferrovales). Fish exhibiting early stage chronic lymphocytic enteritis comprised markedly different global bacterial assemblages compared to those deemed healthy and exhibiting late stages of the disease. This corresponded to an overall loss of diversity and enrichment of Proteobacteria and Actinobacteria, particularly in the gills. In contrast, bacterial assemblages of fish with late stage enteritis were generally similar to those of healthy individuals, though with some distinct taxa. In conclusion, gut health status is an important factor which defines the skin and gill bacterial assemblages of fish and likely reflects changes in immune states and barrier systems during the early onset of conditions like enteritis. This study represents the first to investigate the microbiota of the outer mucosal surfaces of fish in response to underlying chronic gut enteritis, revealing potential biomarkers for assessing fish health in commercial aquaculture systems.

An Economical Custom-Built Drone for Assessing Whale Health

Drones or unmanned Aerial Vehicles’ (UAVs) have huge potential to improve the safety and efficiency of sample collection from wild animals under logistically challenging circumstances. Here we present a method for surveying population health that uses UAVs to sample respiratory vapour, ‘whale blow’, exhaled by free-swimming humpback whales (Megaptera novaeangliae), and coupled this with amplification and sequencing of respiratory tract microbiota. We developed a low-cost multirotor UAV incorporating a sterile petri dish with a remotely operated ‘flip lid’ to sample whale blow with minimal disturbance to the whales. This design addressed several sampling challenges: accessibility; safety; cost, and critically, minimized the collection of atmospheric and seawater microbiota and other potential sources of sample contamination. We collected 59 samples of blow from northward migrating humpback whales off Sydney, Australia and used high throughput sequencing of bacterial ribosomal gene markers to identify putative respiratory tract microbiota. Model-based comparisons with seawater and drone-captured air demonstrated that our system minimized external sources of contamination and successfully captured sufficient material to identify whale blow-specific microbial taxa. Whale-specific taxa included species and genera previously associated with the respiratory tracts or oral cavities of mammals (e.g. Pseudomonas, Clostridia, Cardiobacterium), as well as species previously isolated from dolphin or killer whale blowholes (Corynebacteria, others). Many examples of exogenous marine species were identified, including Tenacibaculum and Psychrobacter spp. that have been associated with the skin microbiota of marine mammals and fish and may include pathogens. This information provides a baseline of respiratory tract microbiota profiles of contemporary whale health. Customized UAVs are a promising new tool for marine megafauna research and may have broad application in cost-effective monitoring and management of whale populations worldwide.

Under control: how a dietary additive can restore the gut microbiome and proteomic profile, and improve disease resilience in a marine teleostean fish fed vegetable diets

BackgroundThe constant increase of aquaculture production and wealthy seafood consumption has forced the industry to explore alternative and more sustainable raw aquafeed materials, and plant ingredients have been used to replace marine feedstuffs in many farmed fish. The objective of the present study was to assess whether plant-based diets can induce changes in the intestinal mucus proteome, gut autochthonous microbiota and disease susceptibility of fish, and whether these changes could be reversed by the addition of sodium butyrate to the diets. Three different trials were performed using the teleostean gilthead sea bream (Sparus aurata) as model. In a first preliminary short-term trial, fish were fed with the additive (0.8%) supplementing a basal diet with low vegetable inclusion (D1) and then challenged with a bacteria to detect possible effects on survival. In a second trial, fish were fed with diets with greater vegetable inclusion levels (D2, D3) and the long-term effect of sodium butyrate at a lower dose (0.4%) added to D3 (D4 diet) was tested on the intestinal proteome and microbiome. In a third trial, the long-term effectiveness of sodium butyrate (D4) to prevent disease outcome after an intestinal parasite (Enteromyxum leei) challenge was tested.ResultsThe results showed that opposed forces were driven by dietary plant ingredients and sodium butyrate supplementation in fish diet. On the one hand, vegetable diets induced high parasite infection levels that provoked drops in growth performance, decreased intestinal microbiota diversity, induced the dominance of the Photobacterium genus, as well as altered the gut mucosal proteome suggesting detrimental effects on intestinal function. On the other hand, butyrate addition slightly decreased cumulative mortality after bacterial challenge, avoided growth retardation in parasitized fish, increased intestinal microbiota diversity with a higher representation of butyrate-producing bacteria and reversed most vegetable diet-induced changes in the gut proteome.ConclusionsThis integrative work gives insights on the pleiotropic effects of a dietary additive on the restoration of intestinal homeostasis and disease resilience, using a multifaceted approach.

Marine Mammal Microbiota Yields Novel Antibiotic with Potent Activity Against Clostridium difficile.

The recent explosion of research on the microbiota has highlighted the important interplay between commensal microorganisms and the health of their cognate hosts. Metabolites isolated from commensal bacteria have been demonstrated to possess a range of antimicrobial activities, and it is widely believed that some of these metabolites modulate host behavior, affecting predisposition to disease and pathogen invasion. Our access to the local marine mammal stranding network and previous successes in mining the fish microbiota poised us to test the hypothesis that the marine mammal microbiota is a novel source of commensal bacteria-produced bioactive metabolites. Examination of intestinal contents from five marine mammals led to the identification of a Micromonospora strain with potent and selective activity against a panel of Gram-positive pathogens and no discernible human cytotoxicity. Compound isolation afforded a new complex glycosylated polyketide, phocoenamicin, with potent activity against the intestinal pathogen Clostridium difficile, an organism challenging to treat in hospital settings. Use of our activity-profiling platform, BioMAP, clustered this metabolite with other known ionophore antibiotics. Fluorescence imaging and flow cytometry confirmed that phocoenamicin is capable of shifting membrane potential without damaging membrane integrity. Thus, exploration of gut microbiota in hosts from diverse environments can serve as a powerful strategy for the discovery of novel antibiotics against human pathogens.

A comparison of recirculation aquaculture systems versus biofloc technology culture system for on-growing of fry of Tinca tinca (Cyprinidae) and fry of grey Mugil cephalus (Mugilidae)

The on-growing of tench Tinca tinca fry (1.81±0.6g) and grey mullet Mugil cephalus fry (0.65±0.2g) was carried out using two different culture systems, recirculation aquaculture system (RAS) and biofloc technology culture system (BFT), to compare their performance and evaluate the technical feasibility of rearing both species using an alternative method. After an on-growing period of 50days, it was possible to verify that the survival rate, fish size in terms of body weight and length, condition factor (K), specific growth rate, gained biomass and apparent feed conversion rate of M. cephalus fry were significantly higher (P<0.05) in RAS in comparison to those obtained using BFT. For Tinca tinca, results were similar for all the measured variables except for the condition factor, that was significantly higher in BFT (P<0.05). Water quality parameters remained within the optimum ranges reported for freshwater fish species using RAS. In BFT, despite the constant addition of glucose, total ammonium concentrations were relatively high (2.89±1.25mg NH4+/L for tench and 3.74±1.34mg NH4+/L for mullet) because the small volume of water in tanks (90L) and the use of inert diets with high protein levels (>50%). Ammonia could only be stabilized when the feed was replaced with one with a lower protein content (35%). The proximate composition of the bioflocs showed that the composition varied according to the fish species considered: with mullet, the protein (17.34±1.40%) and fat (2.36±0.03%) were present in higher concentration than in tench (8.92±0.38% and 2.18±0.18%, respectively), indicating that regardless of the use of the same BFT procedures, bioflocs developing for both species were different. The microbial diversity in the tank water and the intestinal microbiota of fish were examined by restriction fragment length polymorphism (RFLP) and found to be different depending on the system used for on-growing. Thus, in the RAS system the microbial diversity was somewhat higher than in the BFT. As a conclusion, present results indicated that Tinca tinca seem to be able to adapt to the BFT systems.

Supplementation of lactobacilli improves growth, regulates microbiota composition and suppresses skeletal anomalies in juvenile pike-perch (Sander lucioperca) reared in recirculating aquaculture system (RAS): A pilot study

This research aimed to test the effects of lactobacilli, applied to cultured pike-perch, either through hydrolyzed OTOHIME fish diet, or through Artemia nauplii, on fish growth, microbiota balance and skeletal development. On the 12th Day Post Hatching (DPH) fish were divided into following treatment groups: two groups received the combination of OTOHIME and nauplii enriched either with Lactobacillus paracasei BGHN14+Lactobacillus rhamnosus BGT10 or with Lactobacillus reuteri BGGO6–55+Lactobacillus salivarius BGHO1, and one group received OTOHIME hydrolyzed by BGHN14+BGT10 and non-enriched nauplii. Control group received non-enriched nauplii and non-hydrolyzed OTOHIME. The treatment lasted 14days and fish were sacrificed on the 26th DPH for the assessment of digestive enzyme activity and microbiota composition. Individual total lengths and individual body weights were recorded at the end of the treatments, on the 26th DPH, and also on the 45th DPH, in parallel with the evaluation of skeletal deformities and fish survival. Our results indicated positive effect of Artemia enriched with BGGO6–55+BGHO1 on fish growth, skeletal development and trypsin to chymotrypsin activity ratio (T/C), as an indicator of protein digestibility. Hydrolysis of OTOHIME was also associated with better skeletal development, higher T/C values and lower levels of Aeromonas and Mycobacterium spp., which are important fish pathogens. Though additional testing in larger cohort studies is needed, these observations are promising in terms of usage of probiotics for improved environmentally friendly production of pike-perch in Recirculating Aquaculture System (RAS).