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.

We studied the effects of a lactic acid bacterium, Pediococcus acidilactici (strain MA 18/5M, CNCM), as a dietary probiotic on growth performance and some nutritional and microbiological aspects of the shrimp Litopenaeus stylirostris. Experimental shrimp culture was carried out over 10 weeks, using floating cages of 14 m2 each set in earthen ponds, in a New Caledonia farm affected by "Summer syndrome", a septicaemic vibriosis caused by Vibrio nigripulchritudo. The experiment design testing probiotic-coated pellets against control was replicated in two earthen ponds. High mortalities, characteristic of Summer syndrome, were observed during the first 2 weeks of the trial. The probiotic improved production in the treated cages from both ponds with increases in the survival rate (7% and 15% respectively) and final biomass (8% and 12% respectively). No differences were recorded among treatments on growth performances, but significant lower food conversion ratios were obtained with probiotic treatment. After 5 weeks of rearing, the Hepatosomatic Index and the adjusted dry weight of the digestive gland were significantly increased by 10% and by 9% respectively in shrimps treated with probiotic. In the meantime, the specific activities of greek small letter alpha amylase and trypsin in the digestive gland showed a significant effect of the probiotic treatment with respective increases by 35% and 55%. The rise in total trypsin activity following morning feeding was also enhanced by the probiotic treatment (P < 0.001).The highest concentration of probiotic (between 104-105 CFU g− 1 of fresh gut) in the shrimp gut was obtained 2 h after feeding. The concentration remained high for 4 h after feeding before decreasing until the next meal. Total bacterial counts on Marine agar and TCBS in the gut were significantly lowered by the probiotic treatment over the 10 weeks of the experiment. Additionally, during the first 2 weeks, prevalence and load of V. nigripulchritudo strains in haemolymph was lower in animals fed with the probiotic diet.This study demonstrated, under pond conditions, that feeding shrimp with live terrestrial lactic acid bacteria can be an effective treatment for improving shrimp culture affected by vibriosis.

Highlights► Osmoregulation capacity in L. stylirostris is weak in the early life stage. ► Postlarvae progressively acquired the adult pattern of hyper-hypo-osmoregulation. ► At PL9, the estimated isosmotic salinity was 24.5 ppt (720 mOsm.kg − 1 ). ► A two-fold-growth in 19 days-old postlarvae was obtained in dilute seawater compared to seawater conditions.

In New Caledonia, shrimp hatcheries are confronted with mass mortality in the larval stages, a phenomenon poorly understood as no specific causative agent has been identified. This has resulted in an excessive use of prophylactic antibiotics, although their adverse effects in aquaculture are notorious. The present work was thus aimed at selecting potential probiotic strains for penaeid hatcheries. From a pool of more than 400 marine bacterial isolates sampled from the local marine environment seven strains exhibited in vitro antagonistic activity towards Vibrio harveyi. These isolates were characterized both phenotypically and genotypically using a biochemical approach and 16S rDNA sequencing. Six out of these seven strains were found to belong to the genus Pseudoalteromonas, the last one belonging to the Vibrionaceae family and related to the Harveyi clade. Selected probiotic candidates were individually tested for antagonistic activities in vitro using a green fluorescent protein (GFP)-labelled transconjugant of V. harveyi and for inherent pathogenicity towards cultured shrimp larvae at two different developmental stages. This approach enabled rapid processing and selection of candidates to be tested in our experimental hatchery. Four different experiments were conducted to test candidate strains either alone or in combination. Repeated trials showed that postlarval survival was significantly improved by adding the strain NC201 individually to the rearing water compared to unchallenged controls. The analysis of immune-related gene expressions showed that the Litsty PEN3 transcript abundance of larvae was significantly increased after being reared in probiotic-containing water; however no significant difference in lysozyme gene expression was recorded in this study. Together, these results open new insights into the use of these strains as potential substitutes to antibiotherapy in shrimp larval rearing in New Caledonia.