Dietary administration of a commercial mixed-species probiotic improves growth performance and modulates the intestinal immunity of tilapia, Oreochromis niloticus.

The growth performance, immunological status, intestinal morphology and microbiology of tilapia, Oreochromis niloticus, were investigated after dietary administration of the commercial probiotic AquaStar(®) Growout. Tilapia (29.02 ± 0.33 g) were split into five treatments; control (CON), 1.5 g kg(-1) probiotic (PRO-1.5), 3 g kg(-1) probiotic (PRO-3), pulsed probiotic feeding (PRO-PULSE) or an initial probiotic feed followed by control feeding (PRO-INI). After six weeks of experimental feeding, fish fed PRO-3 displayed significantly higher final weight, weight gain and SGR compared to the CON or PRO-INI treatments. Supplementation of the probiotic at this dose induced an up-regulation of intestinal caspase-3, PCNA and HSP70 mRNA levels compared to the CON fed fish. Immuno-modulatory pathways were also affected; significantly higher expression of TLR2, pro-inflammatory genes TNFα and IL-1ß, and anti-inflammatory genes IL-10 and TGFß suggest that the probiotic may potentiate a higher state of mucosal tolerance and immuno-readiness. Histological appraisal revealed significantly higher numbers of intraepithelial leucocytes in the intestine of PRO-3 fed fish compared with treatments CON, PRO-PULSE and PRO-INI but not PRO-1.5. Additionally, fish receiving PRO-3 had a significantly higher abundance of goblet cells in their mid-intestine when compared with fish from all other treatments. Together, these data suggest that continuous provision of AquaStar(®) Growout at 3 g kg(-1) can improve tilapia growth and elevate the intestinal immunological status of the host.

Effects of dietary ß-(1,3)(1,6)-D-glucan supplementation on growth performance, intestinal morphology and haemato-immunological profile of mirror carp (Cyprinus carpio L.).

In recent years, aquaculture research has focused on probiotics, prebiotics, and ß-glucans, in order to improve health status and growth performance. Information regarding the effects of ß-glucan on growth performance and intestinal immunity of mirror carp (Cyprinus carpio L.) is scarce. An experiment was therefore conducted to investigate the effects of a yeast ß-glucan preparation (MacroGard(®) ) on growth performance, intestinal morphology and haemato-immunological indices of mirror carp. Carp (initial weight 11.1 ± 0.0 g) were fed highly purified diets supplemented with 0% (control), 0.1%, 1% or 2% MacroGard(®) for 8 weeks. Fish fed diets containing 1% and 2% MacroGard(®) showed significant improvements in weight gain, specific growth rate and feed conversion ratio compared to fish fed both the control and the 0.1% MacroGard(®) containing diet. Histological appraisal of the intestine showed a significantly higher infiltration of leucocytes into the epithelial layer of fish fed diets supplemented with 1% and 2% MacroGard(®) in the anterior intestine compared to fish fed the control and 0.1% MacroGard(®) diet. This effect was not observed in the posterior intestine. There were no significant differences in the intestinal absorptive surface area and number of goblet cells in either intestinal region. At the end of the experiment, the haematological status of the fish was examined. Compared to control fed fish, the haematocrit value was significantly elevated in fish fed the 2% MacroGard(®) diet. Furthermore, the blood monocyte fraction was significantly higher in fish fed the 1% and 2% MacroGard(®) diets. No significant changes were observed in the other blood parameters assessed. The present study shows that high dietary ß-glucan inclusion increases growth performance without detrimental effects on the health indicators assessed. Increased intraepithelial leucocytes in the anterior intestine may indicate a localized immune response; no detrimental effects on intestinal morphology were observed.

Effects of a dietary ß-(1,3)(1,6)-D-glucan supplementation on intestinal microbial communities and intestinal ultrastructure of mirror carp (Cyprinus carpio L.).

AIM: To assess the effects of dietary Saccharomyces cerevisiae ß-(1,3)(1,6)-D-glucan supplementation (MacroGard(®)) on mirror carp (Cyprinus carpio L.) intestinal microbiota and ultrastructure of the enterocyte apical brush border. METHODS AND RESULTS: Carp were fed either a control diet or diets supplemented with 0.1, 1 or 2% w/w MacroGard(®). Culture-dependent microbiology revealed that aerobic heterotrophic bacterial levels were unaffected by dietary MacroGard(®) after 2 and 4 weeks. No effects were observed on the allochthonous lactic acid bacteria (LAB) populations at either time point; however, reduced autochthonous LAB populations were observed at week 4. PCR-DGGE confirmed these findings through a reduction in the abundance of autochthonous Lactococcus sp. and Vagococcus sp. in MacroGard(®)--fed fish compared with the control-fed fish. Overall, sequence analysis detected microbiota belonging to the phyla Proteobacteria, Firmicutes, Fusobacteria and unidentified uncultured bacteria. DGGE analyses also revealed that dietary MacroGard(®) reduced the number of observed taxonomical units (OTUs) and the species richness of the allochthonous microbiota after 2 weeks, but not after 4 weeks. In contrast, dietary MacroGard(®) reduced the number of OTUs, the species richness and diversity of the autochthonous microbiota after 2 weeks, and those parameters remained reduced after 4 weeks. Transmission electron microscopy revealed that intestinal microvilli length and density were significantly increased after 4 weeks in fish fed diets supplemented with 1% MacroGard(®). CONCLUSIONS: This study indicates that dietary MacroGard(®) supplementation modulates intestinal microbial communities of mirror carp and influences the morphology of the apical brush border. SIGNIFICANCE AND IMPACT OF THE STUDY: To the authors' knowledge, this is the first study to investigate the effects of ß-(1,3)(1,6)-D-glucans on fish gut microbial communities, using culture-independent methods, and the ultrastructure of the apical brush border of the enterocytes in fish. This prebiotic-type effect may help to explain the mechanisms in which ß-glucans provide benefits when fed to fish.

Probiotic Pediococcus acidilactici modulates both localised intestinal- and peripheral-immunity in tilapia (Oreochromis niloticus).

The application of probiotics in aquaculture has received concerted research efforts but the localised intestinal immunological response of fish to probiotic bacteria is poorly understood. Therefore, a study was conducted to evaluate the probiotic effect of Pediococcus acidilactici on Nile tilapia (Oreochromis niloticus) with specific emphasis on intestinal health and probiotic levels as well as system level responses such as growth performance, feed utilization and haemato-immunological parameters under non-challenged conditions. Fish (9.19 ± 0.04 g) were fed either a control diet or a P. acidilactici supplemented diet (at 2.81 × 10(6) CFU g(-)(1)) for six weeks. At the end of the study the probiotic was observed to populate the intestine, accounting for ca. 3% (1.59 × 10(5) CFU g(-)(1)) of the cultivable intestinal bacterial load. Real-time PCR indicated that the probiotic treatment may potentiate the immune-responsiveness of the intestine as up-regulation of the gene expression of the pro-inflammatory cytokine TNFα was observed in the probiotic fed fish (P < 0.05). Light microscopy observations revealed elevated intraepithelial leucocyte (IEL) levels in the intestine of P. acidilactici fed tilapia after six weeks (P < 0.05) of feeding and a trend towards elevated goblet cells was also observed after six weeks feeding (P = 0.08). Concomitantly at week six, along with elevated IELs and elevated TNFα mRNA levels in the intestine, an increased abundance of circulating neutrophils and monocytes were observed in fish fed the probiotic supplemented diet (P < 0.05). This haemopoietic expansion of innate immune cells could be reflective of an elevated state of immuno-readiness. Together these results suggest that the probiotic has a protective action on the intestinal mucosal cells, stimulating the innate immune response after feeding for a period of six weeks. These immunological modulations did not impair growth performance or the remaining haematological and zootechnical parameters compared to the control group (P > 0.05).

Haemato-immunological and growth response of mirror carp (Cyprinus carpio) fed a tropical earthworm meal in experimental diets.

An investigation was conducted to evaluate the effect of feeding a tropical earthworm meal (Perionyx escavatus) on the haemato-immunological response and growth performance of mirror carp (Cyprinus carpio). Fish were fed diets for a total of 88 days, fishmeal served as the main protein source in the control diet. Two remaining diets consisted of fishmeal fixed at 33.65% provision of protein and the remaining 66.35% protein was provided by soybean meal (SBM diet) or P. excavatus meal (EW diet). Compared to control and SBM fed fish (7.69 ± 0.28 and 5.92 ± 0.31 g/dl, respectively), a significant increase in haemoglobin was measured in EW fed fish (9.57 ± 0.24 g/dl). Consequently significant elevations were also observed in mean corpuscular haemoglobin (MCH; 79.13 ± 4.59 pg) and mean corpuscular haemoglobin concentration (MCHC; 22.69 ± 0.54 pg) in EW fed fish. On the contrary, compared to control and SBM fed carp total leukocyte levels (2.72 ± 0.17 and 3.10 ± 0.17 × 10(4)/mm(3), respectively) were significantly decreased in the EW group (2.15 ± 0.14 × 10(4)/mm(3)). Moreover at day 14 and 21 post immunisation with bacterin isolated from Aeromonas hydrophila fish fed the EW diet displayed a significant reduction in respiratory burst activity (RBA) compared to control and SBM fed fish. After 60 days of feeding, fish fed EW diet showed a significant elevation in final body weight compared to fish fed a fishmeal based diet (control treatment) and fish fed a soybean meal based diet. Similar improvements were observed in feed utilisation efficiency. The present study shows that feeding P. excavatus meal to mirror carp decreases some aspects of the innate immune response, but at the same time gives rise to significant enhancement of growth and feed utilisation efficiency.

The effect of Pediococcus acidilactici on the gut microbiota and immune status of on-growing red tilapia (Oreochromis niloticus).

AIM: To assess Pediococcus acidilactici as a dietary supplement for on-growing red tilapia (Oreochromis niloticus). METHODS AND RESULTS: Tilapia were fed either a control diet or control diet supplemented with Ped. acidilactici at 10(7) CFU g(-1) for 32 days. Ped. acidilactici colonized the intestinal tract and significantly affected the intestinal microbial communities. PCR-DGGE revealed direct antagonism of gastric Ped. acidilactici with an endogenous uncultured bacterium during a period of reverting to nonsupplemented feeding. Light microscopy revealed that gut integrity and leucocyte levels were unaffected by Ped. acidilactici; however, blood leucocyte levels and serum lysozyme activity were elevated after 14-days' feeding. No significant improvements in growth performance were observed at the end of the trial (day 32), but survival was significantly higher in the probiotic group. CONCLUSIONS: The study demonstrates that oral supplementation of Ped. acidilactici modulates intestinal bacterial communities in on-growing red tilapia and also stimulates some aspects of the nonspecific immune response. SIGNIFICANCE AND IMPACT OF THE STUDY: To our knowledge this is the first study assessing the effects of probiotics on the gut microbiota of tilapia using culture-independent methods. Such methods are crucial to understand the mechanisms which underpin and mediate host benefits.