Antimicrobial activity in bacteria isolated from Senegalese sole, Solea senegalensis, fed with natural prey

Intestinal bacteria in marine fish may produce antimicrobial substances which inhibit pathogenic bacteria. The aim of this study was to determine the influence of a change of fish diet on the antimicrobial activity of the culturable aerobic gut microflora of Senegalese sole, Solea senegalensis. Pre‐adult 15‐month‐old fish previously fed on an artificial diet, were fed polychaetes (Hediste diversicolor), which form part of the natural diet of Senegalese sole. Samples were taken0, 3 and 6 weeks after start of the experiment from the stomach, small and large intestine of the fish. The bacterial strains isolated from these samples were sub‐cultured to pure cultures and stored at ?80°C. Several biochemical tests were run to obtain some basic phenotypic characteristics of the isolated strains. Amplification and sequencing of 16S rDNA fragments were used to identify the majority of the bacterial strains isolated. The identification by use of this molecular approach gave results in agreement to the phenotypic characterization. Feeding with polychaetes significantly increased (P<0.05) the numbers of presumptive Vibrio isolates in the gut. The number of bacterial strains with antimicrobial activity, as determined by two in vitro approaches, was significantly (P<0.05) increased by feeding with polychaetes.     

Effect of dietary protein level and source on digestive proteolytic enzyme activity in juvenile Senegalese sole, Solea senegalensis Kaup 1850

The effect of dietary protein level and protein source on growth and proteolytic activity of juvenile Solea senegalensis was studied. In Experiment 1, fish were fed on four experimental diets containing increased protein levels (36, 46, 56 and 67%). In Experiment 2, Senegalese soles were fed on five diets with partial substitution of fish meal by soybean meal, soybean protein concentrate, soybean protein isolate, wheat gluten meal or pea protein concentrate. Results prove that growth and proteolytic activity in the distal intestine of fish were affected by the quantitative increase in dietary protein. The origin of protein source used in the elaboration of experimental diets affected both the amount and composition of the alkaline proteases secreted into the intestinal lumen; however, it did not decrease animal growth. Juvenile Senegalese sole showed capability to modulate digestive protease secretion when the concentration and/or source of dietary protein were modified. Quantity and quality of dietary protein affected protein hydrolysis in Senegalese sole intestine. This study establishes that 30% fish meal protein can be replaced by soybean derivatives without affecting intestinal proteases. Replacement with wheat gluten meal or pea protein concentrate should be taken cautiously, but further research is needed to establish whether growth performance and digestive enzyme physiology of Senegalese sole are affected by plant protein-supplemented diets in a long-term trial.     

Structural characteristics and succession of intestinal microbiota for Paralichthys olivaceus during the early life stage

The research of intestinal microflora structures for Paralichthys olivaceus larvae and juveniles will help us to master the ontogeny and developmental colonization of microflora during the larval rearing stage. In this study, we sequenced the total bacterial genomic DNA in larval and juvenile guts with an Illumina MiSeq PE300 system, and analysed the structural characteristics of these microbiota, feed and rearing water in live and formulated feeding periods. The structure of gut microbiota was gradually similar to those in wild P. olivaceus at the phylum level and the newly hatched ones with the growth, according to the distribution and abundance of intestinal microbiota. And the colonized rule of main microbial species in guts was decreased initially and then increased during the larvae and juveniles stage. Meantime, the core microflora of this study were obtained through the analysis of shared and dominant species, which included Bacteroides, Bacillus, Enterococcus, Lactobacillus, Lactococcus, Escherichia_Shigella, Acinetobacter, Pseudomonas, Vibrio, Nitrosomonas, and Glaciecola. The correlation analysis of microbiota between intestines and environmental factors suggested that microflora in feed and water could affect the distribution of larval and juvenile gut microbiota. Moreover, many species of Acinetobacter, Pseudomonas and Vibrio are treated as important potential pathogens in aquaculture. These all pointed out the microbial quality of feed and rearing water should be strictly controlled in fish breeding and farming, and supply theoretical bases for screening the native probiotics to artificially regulate the gut microbiota.     

Influence of dietary administration of a probiotic strain Shewanella putrefaciens on senegalese sole (Solea senegalensis,Kaup 1858) growth,body composition and resistance to Photobacterium damselae subsp piscicida

Probiotics are currently being supplemented to cultured fish due to their benefits for fish performance. Herein, we tested the health protection and nutritional effects of probiotic Shewanella putrefaciens (Pdp11 strain), both fresh and lyophilized cells, on Senegalese sole (Solea senegalensis) juveniles. Pdp11 was incorporated into fish feed at concentration of 10⁹ cells g^?1, and then provided to juvenile sole for 2 months. Growth rates were significantly higher in fish fed the fresh probiotic compared with fish fed the control diet. Growth of fish receiving lyophilized bacteria was not improved when compared with controls. Body compositions (protein, total lipids and fatty acids profile) were similar for each dietary treatment. At the end of the study, fish were challenged by intraperitoneal inoculation with a pathogenic strain of Photobacterium damselae subsp. piscicida. Relative percentages of survival (RPS) ranged between 25% and 43.8% for fish receiving both probiotic supplemented diets. Both fresh and lyophilized Pdp11 cells conferred protection against P. damselae subsp. piscicida. However, only fresh Pdp11 enhanced Senegal sole performance. These findings should be taken into account for industry purposes in which lyophilized Pdp11 might be easier to manage.     

Immune responses and gut morphology in Senegalese sole (Solea senegalensis) fed dietary probiotic supplementation and following exposure to Photobacterium damselae subsp. piscicida

The aim of this study was to determine the effect of a dietary multi‐species probiotic on growth, gut morphology and immune parameters in Senegalese sole (Solea senegalensis). Fish were fed with two experimental diets, a control diet and the same diet supplemented with the probiotic for 72 days. A sub‐lethal bath challenge with Photobacterium damselae subsp. piscicida was performed after the growth trial. Intestine and blood samples were collected to study gut structure and plasmatic immune parameters. No significant differences were found in growth performance. The analysis of gut morphology showed a significant increase in intestinal villi height of Senegalese sole fed the probiotic. Regarding circulating leucocytes, dietary probiotic supplementation increased thrombocytes levels whereas a decrease in the proportion of lymphocytes was observed. No significant differences were observed in humoural immune parameters. Bath challenge differentially affected leucocyte counts and increased peroxidase activity. This study presents the possibility of using dietary probiotic supplementation to increase Senegalese sole welfare since gut morphology was positively affected. Although the immune response after a bacterial challenge was not modulated by dietary treatment, further studies would be instrumental to unravel eventual dietary benefits on immune mechanisms.     

Microbiota of yellow grouper (Epinephelus awoora Temminck & Schlegel, 1842) fed two different diets

The rapidly growing yellow grouper industry has experienced relatively severe bacterial disease problems in China. The proliferation of pathogens in fish can be suppressed by commensal microbiota. In this background, we used nested polymerase chain reaction‐denaturing gradient gel electrophoresis (PCR‐DGGE) and sequence analysis to investigate microbiota in the skin, gills and intestines, including adherent bacteria and non‐adherent bacteria in yellow grouper fed with natural diet and complete feed. A total of 21 bacterial species were identified using phylogenetic analysis. The γ‐Proteobacteria group (81.0%, 17 species) dominated the bacterial communities in yellow grouper completely. Others belonged to Firmicutes (9.5%, two species), Actinobacteria (4.75%, one species) and Verrucomicrobia (4.75%, one species). The higher similarities (above 91%) of the DGGE band patterns in skin, gill and intestinal‐non‐adherent bacteria between two groups of fish indicated that existed more stable microbial communities existed in these specifically ecological niches in yellow grouper. However, considerable differences existed between two intestinal‐adherent bacteria (IAB) samples; that is, compared with natural diet fed yellow grouper, higher bacterial apparent species richness and possibly less abundance existed in IAB in fish fed with complete diets, probably indicating that the community structures in IAB were affected easily and significantly by diet.     

Tissue composition,blood biochemistry and histology of digestive organs in Senegalese sole (Solea senegalensis) juveniles fed diets containing different plant protein ingredients

The aim of this study was to determine the impact of diets with different plant protein ingredients on proximate composition, tissue metabolites and tissue fatty acid composition, as well as hepatic and intestinal histological modifications in Senegalese sole (Solea senegalensis). Fish (21.5 ± 2.8 g body weight) were fed six isonitrogenous and isoenergetic diets during 11 weeks. A control diet containing fish meal as the main protein source was compared with five experimental diets replacing 30% fish meal protein by different plant protein sources: soybean meal (SBM), soybean protein concentrate (SPC), soybean protein isolate (SPI), wheat gluten meal (WGM) or pea protein concentrate (PPC). The inclusion of different plant protein did not significantly affect growth and proximate composition of fish. The hepatosomatic index was not significantly different to the control group; however, utilization of WGM significantly increased hepatocyte size. Plasma protein values and muscle triglycerides were influenced by the inclusion of SBM and WGM in the diets respectively. Feeding fish on SBM, WGM and PPC diets significantly enhanced n‐6 fatty acid levels in muscle, particularly linoleic acid. None of the plant protein ingredient used in the diets decreased arachidonic, eicosapentaenoic as well as docosahexaenoic acid values in liver and muscle. Overall, histological studies did not reveal the existence of any intestinal alterations induced by the inclusion of different plant proteins. Despite moderate changes produced by SBM, SPC and WGM, inclusion of dietary plant protein ingredients has no major impact on growth, tissue and blood biochemistry, fatty acid profile and gut integrity of Senegalese sole juveniles.     

Evaluation of Feed Attractants in Juvenile Senegalese Sole,Solea senegalensis

The Senegalese sole, Solea senegalensis, is a flatfish with increasing interest in the European aquaculture. However, one of the main limitations found in this species is related to the scarce attraction to commercial aquafeeds, thus determining low feed intake in fish farms. The aim of this study was to evaluate four ingredients of marine origin (fish meal hydrolysate [FMH], polychaete meal [POM], mussel meal [MUM], and squid meal [SQM]) as potential attractants for juvenile Senegalese sole. Four experimental feeds containing 10% of the potential attractants, plus a control diet without these ingredients, were evaluated. A total of 150 fish were distributed into ten 40‐L tanks (duplicate tanks per feed). A Latin square design was carried out, and hence all the experimental diets were offered to each tank in successive periods of 7 d. Fish were fed twice a day (morning and afternoon), and the feeding behavior of the animals was assessed by direct observations. Behavioral pattern assessed was classified as distant orientation, near orientation, or continuation of intake. The acceptability index for each behavioral phase and experimental feeds used was established for each observation. No behavioral differences could be noticed between the morning and the afternoon feedings. FMH showed the greatest attractant potential than among the ingredients assessed, and consequently this ingredient yielded the higher feed intake rates. On the other hand, POM was not the best attractant for juvenile Senegalese sole.     

The autochthonous microbiota of the freshwater omnivores jundiá (Rhamdia quelen) and tilapia (Oreochromis niloticus) and the effect of dietary carbohydrates

Previous studies have shown that the gut microbiota of fish differs depending on host feeding habits and these populations may also be influenced by dietary ingredients. In this study, the bacterial populations of the intestinal tract of the opportunistic omnivore jundiá Rhamdia quelen and the typical omnivore tilapia Oreochromis niloticus were investigated in two experiments. In experiment I, the levels of amylolytic, cellulolytic, lipolytic, proteolytic and total culturable bacteria were compared in the proximal, mid and distal intestine using selective agar. Higher levels of amylolytic, cellulolytic, lipolytic, proteolytic and total culturable bacteria were observed in the tilapia than jundiá, in all intestinal regions. Jundiá contained higher levels of proteolytic bacteria and lipid digesting bacteria in the distal intestinal portion as compared with the mid intestine; both fish species also presented more amylolytic bacteria in the distal intestine. In experiment II, the amylolytic intestinal microbiota between the two species was compared after administration of diets containing different carbohydrate sources. Jundiá fed broken rice presented higher total culturable bacterial levels; however, dietary cassava bagasse and ground corn significantly elevated the population of amylolytic bacteria in tilapia (P > 0.05). PCR‐DGGE was also used to assess the bacterial communities in experiment II. A Cetobacterium spp. was detected in jundiá fed diets containing broken rice, and tilapia fed cassava bagasse, dextrin, broken rice and ground corn. Microbial differentiation was further demonstrated between jundiá and tilapia, because an uncultured bacterium was unique in tilapia and an uncultured spirochete was observed only in jundiá; the presence of these bacterial species was also influenced by dietary carbohydrate sources.     

Effects of dietary orange peel on growth performance,antioxidant activity,intestinal microbiota and liver histology of Gilthead sea bream (Sparus aurata) larvae

This study investigated the effects of dietary orange peel (OP) on growth performance, feed utilization, antioxidant activity, intestinal microbiota and liver histology of Gilthead sea bream (Sparus aurata) larvae (0.32 ± 0.01 g/fish) (mean ± SD). Fish fed iso‐nitrogenous (480 g/kg protein) and iso‐energetic (23 MJ/kg) diets supplemented with OP at concentrations of 0, 1, 3 or 5 g/kg diet, for 60 days. Growth performance and feed utilization parameters were significantly improved by the elevation of dietary OP level, and the optimum level was 5 g/kg diet. The maximum activity of the total antioxidant capacity, superoxide dismutase, glutathione peroxidase, catalase, alkaline phosphatase enzyme and malondialdehyde in the liver was found at 5.5, 4.6, 3.4, 2.9, 3.7 and 3.8 g OP/kg diet, respectively. All tested bacteria (Staphylococcus spp., Vibrio spp. and Salmonela spp) and total bacterial count decreased significantly in the gut of fish fed high levels of OP (3.0 or 5.0 g/kg). No differences were found in the liver histo‐architecture among treatments after 60 days feeding on OP diets. In summary, dietary OP improved growth rate, antioxidant activity and intestinal microbiota of S. aurata larvae with an optimum range from 2.9 to 5.5 g/kg diet.     

Intestinal microbiota of mangrove red snapper (Lutjanus argentimaculatus Forsskål, 1775) reared in sea cages

Using polymerase chain reaction amplification of 16S rDNA coupled to denaturing gradient gel electrophoresis (DGGE) and sequencing of isolated amplicons, we investigated the microbiota of the intestinal digesta and mucosal surface in mangrove red snapper cultured in a cage aquaculture area in Daya Bay. A total of 14 sequences were characterized by phylogenetic analysis. Among the bacterial species determinated from sequences, the γ‐Proteobacteria group (64.25%, nine species) dominated absolutely in fish intestines. Others belonged to Spirochaetes (14.3%, two species), Cyanobacteria (14.3%, two species) and Firmicutes (7.15%, one species). However, the bacteria were identified as uncultured accounting for 28.6% (four species). The apparent bacterial richness (calculated as the numbers of DGGE bands) was significantly higher in digesta than that in mucosal tissue samples (P<0.05). There existed five dominant individual populations including one unknown species of Firmicutes, Arthrospira sp., Vibrio metschnikovii, Vibrio harveyi and Vibrio sp. in intestinal digesta, and in contrast, only three dominant individual populations including Vibrio natriegens, V. harveyi and Vibrio sp. in intestinal mucosal surface. The results indicated that the microbiota in intestinal digesta was significantly different from that in mucosal surface.     

Effect of dietary carbohydrate-to-lipid ratios on growth, lipid deposition and metabolic hepatic enzymes in juvenile Senegalese sole (Solea senegalensis, Kaup)

A study was undertaken to determine the effect of various dietary carbohydrate‐to‐lipid ratios on growth performance, whole‐body composition and tissue lipid content in Senegalese sole (Solea senegalensis) juveniles. Data on the dietary regulation of key hepatic enzymes of the lipogenic and glycolytic pathways (glucose‐6‐phosphate dehydrogenase, G6PD; malic enzyme, ME; fatty acid synthetase, FAS; pyruvate kinase, PK and glucokinase, GK) were also generated. Four isonitrogenous (crude protein: 52% dry matter (DM)) diets were formulated to contain one of two lipid levels (11% and 21% DM). Within each dietary lipid level, the nature of the carbohydrate fraction (raw or extruded peas) was varied. Triplicate groups of 54 sole (initial body weight: 23.6±1.2 g) were grown in recirculated seawater over 67 days. Fish were fed using automated feeders. At the end of the study, whole‐body, liver, viscera and muscle samples were withdrawn for analyses. During the experimental period, the mean fish weight about doubled in all treatments. No significant differences were found in growth performance (ranging from 1.1% to 1.4% body weight day^?1) among dietary treatments. High‐fat diets increased whole‐body fat content. Similarly, daily fat gain ranged from 0.54 to 0.78 g kg^?1 day^?1 and highest values were found in fish fed high‐lipid diets. Dietary treatments also affected tissue lipid content (liver, viscera and muscle), with highest values in fish fed high‐fat diets. The nature of dietary carbohydrates had little influence on performance criteria, but affected tissue lipid deposition. The activities of G6PD, ME and FAS were depressed by elevated levels of dietary lipid, confirming the inhibitory effect of dietary fats on lipid biosynthesis. At both dietary lipid levels, ME and FAS activities were little affected by dietary carbohydrate. Activities of PK and GK were not affected by the starch level of the diets. In Senegalese sole juveniles, the lipogenic pathway is more susceptible to modulation by dietary means (particularly through lipid intake) than the glycolytic pathway.     

Senegalese sole larvae growth and protein utilization is depressed when co‐fed high levels of inert diet and Artemia since first feeding

A large effort has been dedicated in the past years to the development of nutritional balanced inert diets for marine fish larvae in order to suppress the nutritional deficiencies of live feed. In this study growth performance, Artemia intake, protein digestibility and protein retention were measured for Senegalese sole (Solea senegalensis Kaup), in order to provide insight into how protein utilization affects growth performance. Three feeding regimes were tested: ST – standard live feed; ArtRL – live feed and 20%Artemia replacement with inert diet (dry matter basis) from mouth opening; ArtRH – live feed and 58%Artemia replacement with inert diet from mouth opening. Artemia intake and protein metabolism were determined at 6, 15 and 21 days after hatching using ¹⁴C‐labelled Artemia protein and subsequent incubation in metabolic chambers. At the end of the experiment, sole fed exclusively with live feed were significantly larger than sole from Artemia replacement treatments. Protein digestibility decreased during sole ontogeny, and more sharply in ArtRH sole. Concomitantly retention efficiency increased during ontogeny but with a slight delay in ArtRH sole. Senegalese sole larvae growth and protein utilization is depressed when co‐fed high levels of inert diet and Artemia, mostly during metamorphosis climax.     

Effects of dietary copper sources on growth,tissue copper accumulation and physiological responses of Japanese sea bass (Lateolabrax japonicus) (Cuvier, 1828) fed semipurified or practical diets

An 8‐week trial was conducted to compare the efficacy of a chelated copper (Cu) with the inorganic Cu (CuSO₄), in the diet of Japanese sea bass (Lateolabrax japonicus). Six diets were prepared based on two basal diets (semi‐purified or practical) at two Cu inclusion levels (0 and 4 mg kg^?1 from or chelated‐Cu), accordingly named as S0, SI4, SM4, P0, PI4 and PM4 respectively. Each diet was fed to quadruplicate groups of Japanese sea bass (initial weight: 65.0 ± 0.03 g) during the trial. Fish fed practical diets had higher feed intake, specific growth rate, protein efficiency rate (PER), liver Cu accumulation, liver Na⁺K⁺‐ATPase and plasma alkaline phosphatase, but lower hepatosomatic index than the groups fed semi‐purified diets (P < 0.05). Higher PER was exhibited by fish fed chelated‐Cu diets than that of fish fed CuSO₄ diets (P < 0.05). Although liver Cu accumulation was significantly higher in fish fed practical diets,, liver Cu content increased significantly with increasing dietary copper levels only in fish fed semipurified diets. Chelated‐Cu has higher efficiency than CuSO₄ when fed in practical diets. Poor growth performance of fish fed semipurified diets might induce underestimating the nutrient requirement of Japanese sea bass.     

Soy protein concentrate as a protein source for Senegalese sole (Solea senegalensis Kaup 1858) diets: effects on growth and amino acid metabolism of postlarvae

The objective of this work was to evaluate the effect of a dietary amino acid imbalance, originating from the use of a soy protein concentrate (SPC) as the major protein source, on the growth performance and amino acid metabolism of Senegalese sole (Solea senegalensis) postlarvae. Senegalese sole (85.6±24.6 mg wet weight) were fed one of two experimental diets: one based on fish meal (FM) and another based on SPC. Diets were isonitrogenous (around 56% crude protein) and isoenergetic. Diet acceptability was very good and the growth rate was 6.9% day^?1 for sole eating the FM diet and 6.0% day^?1 for sole eating the SPC diet. Mass‐specific ammonia excretion and the activities of selected amino acid metabolic enzymes (ALAT, ASAT and GDH) did not present significant differences between treatments, although this may have been due to the high variability found for these parameters in the SPC treatment. This variability may suggest different capacities of individual fish to adapt to the possible methionine dietary deficiency. The utilization of amino acids as a substrate for lipogenesis does not seem to be affected by the dietary protein source, since NAPDH‐generating enzymes (G6PD and ME) had similar activities in both treatments. Amino acid metabolism in Senegalese sole postlarvae seems to be slightly affected by the dietary protein source. Nevertheless, the changes induced by the SPC diet do not seem to impair growth, at least at the high dietary protein level used in this experiment.     

Optimal dietary protein levels in juvenile Senegalese sole (Solea senegalensis)

A study was undertaken to determine the dietary protein level for optimal growth performance and body composition of juvenile Senegalese sole. Five experimental extruded diets were formulated to contain increasing levels of protein [430, 480, 530, 570 and 600 g kg^?1 dry matter (DM)] and a constant lipid level, ranging from 100 to 130 g kg^?1 DM. Triplicate groups of 35 sole (initial body weight: 11.9 ± 0.5 g) were grown over 84 days in 60‐L tanks supplied with recirculated seawater. Fish were fed by means of automatic feeders in eight meals per day. At the start and end of the trial, whole‐body samples were withdrawn for proximate composition analysis. At the end of 84 days of experimental feeding, daily weight gain and specific growth rate in fish fed diets P43 and P48 were significantly lower than those found in fish fed higher protein level diets (P53, P57 and P59). Similarly, feed efficiency was also significantly lower in fish fed diet P43 than in fish fed all other dietary treatments. Sole juveniles fed lower protein level diets (P43 and P48) showed a significantly lower protein content than fish fed the higher dietary protein level treatments (P53, P57 and P60). Changes within the tested dietary protein levels did not affect significantly protein productive value or total nitrogen (N) losses in fish. However, daily N gain was significantly higher (P < 0.05) in fish fed diets P53 and P60 than in fish fed the lowest protein level diet (P43). Data from the present study indicate that diets for juvenile Senegalese sole should include at least 53% crude protein to maintain a good overall growth performance. Based on a second‐order polynomial regression model, the daily crude protein requirement for maximum whole‐body N gain as estimated here for Senegalese sole juveniles was 6.43 g kg^?1 body weight day^?1 which corresponds to a value of 1.03 g N intake kg^?1 body weight day^?1. If the present data are expressed on a dietary crude protein concentration basis, the allowance for maximum protein accretion (N gain) would be met by a diet containing a crude protein level of 600 g kg^?1.     

Bacterial community of pond's water,sediment and in the guts of tilapia (Oreochromis niloticus) juveniles characterized by fluorescent in situ hybridization technique

Information about bacterial community structure and functioning in fish farming ponds remains scarce, mainly due to methodological difficulties in counting and identifying uncultured bacteria. The main objective of this study was to evaluate the degree of similarity between the bacterial community of the digestive tract of tilapia (Oreochromis niloticus) juveniles and that of the test pond's water and sediment, using the Fluorescent In Situ Hybridization (FISH) technique. Samples of water, sediment and gut content of 30 tilapia juveniles from a single nursery ground were collected in January 2010. Potentially probiotic and pathogenic bacteria of the species Bacillus, Lactobacillus brevis, Lactobacillus collinoides, Lactobacillus coryniformis, Lactobacillus farciminis, Vibrio and Pseudomonas fluorescens were found in different samples using specific fluorescent probes. The similarity between bacterial community environments and gastrointestinal tracts was determined using the Morisita‐Horn index. The fish guts presented higher abundances of Pseudomonas fluorescens, Bacillus, Lactobacillus brevis and Lactobacillus collinoides. The bacterial community composition of tilapia gastrointestinal tract was more similar to the water than the sediment of the pond. The results of this study showed that the FISH technique can be easily used for monitoring of probiotics and pathogen detection in aquaculture systems.     

Influence of the dietary inclusion of <Emphasis Type="Italic">Gracilaria cornea</Emphasis> and <Emphasis Type="Italic">Ulva rigida</Emphasis> on the biodiversity of the intestinal microbiota of <Emphasis Type="Italic">Sparus aurata</Emphasis> juveniles

Partial substitutions of fish meal by 5, 15, or 25 % of Gracilaria cornea or Ulva rigida in experimental diets were evaluated to study their effects on biodiversity of intestinal microbiota composition in gilthead seabream (Sparus aurata) juveniles. The diets were offered to duplicate groups of 15 juvenile fish (14.0 ± 0.5 g) for 70 days, and at the end of the experiment the intestinal microbiota from four specimens of each treatment was analysed by denaturing gel gradient electrophoresis. Results showed that the substitution of fish meal by algae meal induced important modifications in the intestinal microbiota community, as a big reduction of the biodiversity when the highest percentage (25 %) of U. rigida was included. On the contrary, an increase on the number of species was detected when a 15 % of algae was included. Various Lactobacillus delbrueckii subspecies were selectively stimulated when G. cornea was included in the feed, and other bacterial species, such as those included in the Vibrio genus, were reduced.     

Effect of dietary supplementation of inulin and W. cibaria on haemato‐immunological parameters of hybrid surubim (Pseudoplatystoma sp)

The dietary supplementation of prebiotics, probiotics and symbiotic in hybrid surubins (a Pseudoplatystoma corruscans and P. fasciatum cross) was evaluated for the effects on their autochthonous intestinal microbiota and on haematological and immunological parameters. A total of 160 fish were divided into four treatment groups with four replicates each. The treatment groups were fed with the following diets for 15 days: control diet without supplementation; 0.5% inulin (prebiotic) supplementation; Weissella cibaria (CPQBA 001‐10 DRM 02) (7.87 ± 0.2 log CFU g^?1) supplementation; or 0.5% inulin and W. cibaria supplementation (symbiotic group). The midgut intestines of the fish with the symbiotic diet supplementation had higher concentrations of lactic acid bacteria (7.07 ± 1.11 log CFU g^?1) and low levels of Vibrio spp (1.90 ± 0.60 log CFU g^?1) and Pseudomonas spp (2.23 ± 1.48 log CFU g^?1). In addition, increased erythrocytes and reduced circulating neutrophils were observed in this group. No differences in blood glucose, serum protein or lysozyme levels were detected between treatment groups. However, a higher concentration of total immunoglobulin was observed in fish fed with the probiotic and symbiotic diets. The addition of 0.5% inulin (prebiotic) thus W. cibaria (probiotic) to the diet of Pseudoplatystoma hybrid surubins reduce the number of pathogenic bacteria and stimulate the beneficial intestinal microbiota and may possibly alter their immune defence system.     

Interactive effects of dietary protein and live bakery yeast, Saccharomyces cerevisiae on growth performance of Nile tilapia, Oreochromis niloticus (L.) fry and their challenge against Aeromonas hydrophila infection

Yeasts used as a probiotic in fish diets could stimulate fish resistance against bacterial infection and could enhance the activities of digestive enzymes in fish guts. In addition to yeast importance, dietary protein is another important part in fish diets that should be carefully optimized to meet fish requirement. It is proposed that the yeast supplementation may enhance the dietary protein turnover and reduce the protein requirement for fish. Therefore, the interactive effects of dietary protein and yeast levels on the growth performance of Nile tilapia, Oreochromis niloticus (L.) fry and their challenge against Aeromonas hydrophila infection was evaluated. In the present study, ten experimental diets were formulated to contain either 35% or 45% crude protein (CP). For each protein level treatment, bakery yeast (Saccharomyces cerevisiae) was supplemented at 0.0, 0.50, 1.0, 2.0, or 5.0 g/kg diet. Fish (0.25–0.48 g) were distributed at a rate of 25 fish per 140-L aquarium. For each diet, triplicate aquaria were fed twice a day, 5 days a week for 12 weeks. Fish growth and feed utilization were significantly affected by either dietary protein or yeast levels alone, while no significant effect of their interaction was observed. The highest fish growth was obtained at 1.0–5.0 g yeast/kg diet at both protein levels; however, the fish performance at 45% CP was better than that fed on 35% CP diets. The optimum feed conversion ratio (FCR) was obtained when fish fed on 1.0–5.0 and 2.0–5.0 g yeast/kg diet at 35 and 45% CP, respectively. The cumulative fish mortality, after interperitoneal injection with A. hydrophila for 10 days, and bactericidal activity was significantly higher in fish fed 35% CP diets than those fed 45% CP diets. Both variables decreased significantly with the increase in yeast levels. The lowest bacterial count and bactericidal activity were obtained in fish fed 5.0 g yeast/kg diet irrespective to dietary protein levels. It could be concluded that the inclusion of live bakery yeast in practical diets could improve the growth performances, feed utilization, and physiological status of Nile tilapia fry and their challenge against A. hydrophila infection. Moreover, fish performance when fed 45% CP diet was better than those fed 35% CP diet. Based on these results, the most suitable yeast level for maximum Nile tilapia growth was determined to be 2.0 g yeast/kg diet with 45% CP diet; however, this level was recommended to stimulate their productive performance and enhances their resistance against A. hydrophila infection.