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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).

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Antioxidant defences and induced oxidative stress tissue damage of the blue shrimp Litopenaeus stylirostris, under challenge with Vibrio nigripulchritudo, were investigated for a 72-h period. For this purpose, L. stylirostris were first infected by immersion with pathogenic V. nigripulchritudo strain SFn1 and then antioxidant defences: superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (Gpx), Total antioxidant status (TAS), glutathiones and induced tissue damage (MDA and carbonyl proteins) were determined in the digestive gland at 0, 12, 24, 48 and 72 h post-infection (h.p.i.). In the meantime, TAS was also measured in the blood. Infection level of the shrimps during the challenge was followed by determining V. nigripulchritudo prevalence and load in the haemolymph of the shrimps. Changes in all these parameters during the 72-h.p.i. period were recorded for control shrimps and shrimps previously fed for one month with probiotic Pediococcus acidilactici MA18/5M at 10(7) CFU g(-1) of feed. Our results showed that immersion with V. nigripulchritudo led to maximal infection level in the haemolymph at 24 h.p.i. preceding the mortality peak recorded at 48 h.p.i. Significant decreases in the antioxidant defences were detected from 24 h.p.i. and beyond that time infection leaded to increases in oxidative stress level and tissue damage. Compared to control group, shrimps fed the probiotic diet showed lower infection (20% instead of 45% at 24 h.p.i. in the control group) and mortality (25% instead of 41.7% in the control group) levels. Moreover, infected shrimp fed the probiotic compared to uninfected control shrimps exhibited very similar antioxidant status and oxidative stress level. Compared to the infected control group, shrimps fed the probiotic sustained higher antioxidant defences and lower oxidative stress level. This study shows that bacterial infection leads to oxidative stress in L. stylirostris and highlighted a beneficial effect of P. acidilactici, suggesting both a competitive exclusion effect leading to a reduction of the infection level and/or an enhancement of the antioxidant status of the shrimps.

In this study we evaluated the effects of the probiotic Pediococcus acidilactici MA18/5M on shrimp, Litopenaeus stylirostris (also called Penaeus stylirostris), first on antioxidant defences and secondly on the oxidative stress status in the shrimps' haemolymph and digestive gland. We conducted two experiments with the same protocol in which shrimps were fed two diets for three weeks: a control diet and a probiotic diet containing 1 g of live P. acidilactici MA18/5M kg − 1. In the first experiment, the shrimps were found to be healthy over the trial period; no mortalities and nondetectable signs of infection were recorded. These resulted in high final survival rates (above 90% in both treatments). On the other hand, during the second trial, carried out at a period of increased risk for an outbreak of the summer syndrome, higher mortalities were recorded, associated with high V. nigripulchritudo prevalence and loads in the shrimp haemolymph. In healthy shrimps (trial 1), no detectable response of the antioxidant defence system and of oxidative stress bio-indicators were observed. However, feeding the probiotic significantly increased the total antioxidant status (TAS) and glutathione peroxidase activity, while all other parameters remained significantly unchanged. In the shrimps exposed to V. nigripulchritudo (trial 2), the antioxidant response was characterized by higher antioxidant enzyme activities (superoxide dismutase and catalase) and higher oxidative stress level in the digestive gland (higher oxidized/reduced glutathione ratio, higher malondialdehyde and carbonyl protein contents) compared to levels found in trial 1. However, shrimps fed the probiotic diet exhibited (i) significantly higher final survival rates (67 ± 3%) compared to the control (47 ± 4%) and (ii) a lower prevalence of V. nigripulchritudo throughout the trial. Moreover, the antioxidant response and the oxidative stress level recorded in the digestive gland with shrimps submitted to the probiotic diet were lower. In the light of these results, we confirm that P. acidilactici enrichment in shrimps' diet seems an effective way of reducing the susceptibility of shrimps to bacterial pathogens.