Dietary administration of a postbiotic, heat-killed Pediococcus pentosaceus PP4012 enhances growth performance, immune response and modulates intestinal microbiota of white shrimp, Penaeus vannamei.

The efficacy of postbiotics on the immune-related gene expression and gut microbiota of white shrimp, Penaeus vannamei remains unexplored. A commercial heat-killed postbiotic Pediococcus pentosaceus PP4012 was used to evaluate the growth performance, intestinal morphology, immunological status, and microbial community of white shrimp after dietary administration in this study. White shrimp (0.040 ± 0.003 g) were divided into three treatments; a control, inanimate P. pentosaceus (105 CFU g feed-1) at low concentration (IPL) and inanimate P. pentosaceus (106 CFU g feed-1) at high concentrations (IPH). The diets of IPL and IPH significantly increased final weight, specific growth rate and production compared to the control group. Shrimp fed with IPL and IPH significantly utilized feed more efficiently than those fed the control diet. The IPH treatment significantly lowered the cumulative mortality rate compared to the control and IPL diet following Vibrio parahaemolyticus infection. No significant difference was observed for Vibrio-like and lactic acid bacteria in intestine of shrimp fed with the control diet and the experimental diets. Adding inanimate P. pentosaceus significantly improved immune responses such as lysozyme and phagocytic activity compared to the control group. However, the total hemocyte count, phenoloxidase activity, respiratory burst, and superoxide dismutase activity were not significantly different among treatments. The immune-related genes alf, pen3a, and pen4 expression were significantly higher in shrimp fed IPL diet compared with control and IPH. Taxonomic identification of bacterial genera in all dietary groups belonged to two predominant phyla, Proteobacteria and Bacteroidota. An abundance of Photobacterium, Motilimonas, Litorilituus, and Firmicutes bacterium ZOR0006 were identified in the intestine of shrimp fed postbiotic diets. Unique microbes such as Cohaesibacter was discovered in the shrimp fed IPL while Candidatus Campbellbacteria, uncultured Verrucomicrobium DEV114 and Paenalcaligenes were discovered in the intestines of shrimp fed IPH diet. Collectively, these data suggest that including heat-killed P. pentosaceus, particularly IPH, can enhance growth performance, promote microbial diversity, elevate immune responses, and increase shrimp's resistance to V. parahaemolyticus.

Ballantyne R ，Lee JW ，Wang ST ，Lin JS ，Tseng DY ，Liao YC ，Chang HT ，Lee TY ，Liu CH ... -  《-》

A comparative study on effects of dietary three strains of lactic acid bacteria on the growth performance, immune responses, disease resistance and intestinal microbiota of Pacific white shrimp, Penaeus vannamei.

The present study evaluated the growth performance, immune responses, disease resistance and intestinal microbiota in Penaeus vannamei fed diets supplemented with three strains of lactic acid bacteria (LAB). The basal diet (control, CO) supplemented with Lactobacillus plantarum W2 (LA), Pediococcus acidilactici Nj (PE), Enterococcus faecium LYB (EN) and florfenicol (FL), respectively, formed three LAB diets (1 × 1010 cfu kg-1) and a florfenicol diet (15 mg kg-1, positive control), were fed to shrimp for 42 days. Results indicated that specific growth rate, feed efficiency rate, and disease resistance of shrimp against Vibrio parahaemolyticus in the treatment groups were significantly improved versus the control (P < 0.05). Compared with the control, acid phosphatase, alkaline phosphatase, phenonoloxidase, total nitric oxide synthase, peroxidase, superoxide dismutase activities, total antioxidant capacity, and lysozyme content in the serum and the relative expression levels of SOD, LZM, proPO, LGBP, HSP70, Imd, Toll, Relish, TOR, 4E-BP, eIF4E1α and eIF4E2 genes in the hepatopancreas of LAB groups were enhanced to various extents. Intestinal microbiota analysis showed that the LA and EN groups significantly improved microbial diversity and richness, and LAB groups significantly altered intestinal microbial structure of shrimp. At the phylum level, the Verrucomicrobiota in the LA and PE groups, the Firmicutes in the EN group, and the Actinobacteriota in the PE and EN groups were enriched. Moreover, the CO group increased the proportion of potential pathogens (Vibrionaceae and Flavobacteriaceae). The potential pathogen (Vibrio) was reduced, and potential beneficial bacteria (Tenacibaculum, Ruegeria and Bdellovibrio) were enriched in response to dietary three strains of LAB. When the intestinal microbiota homeostasis of shrimp is considered, L. plantarum and E. faecium showed better effects than P. acidilactici. However, due to the concerns on the possible potential risks of E. faecium strains to human health, L. plantarum W2 is more suitable for application in aquaculture than E. faecium LYB. Considering collectively the above, Lactobacillus plantarum W2 could be applied as better probiotic to improve the growth performance, non-specific immunity, disease resistance and promote intestinal health of P. vannamei.

Luo K ，Liu Y ，Qin G ，Wang S ，Wei C ，Pan M ，Guo Z ，Liu Q ，Tian X ... -  《-》

Effect of dietary supplementation with Moringa oleifera leaf extract and Lactobacillus acidophilus on growth performance, intestinal microbiota, immune response, and disease resistance in whiteleg shrimp (Penaeus vannamei).

This study investigated the effect of the moringa (Moringa oleifera) leaf extract and Lactobacillus acidophilus individually or combined on growth performance, enzyme activity, intestinal and hepatopancreatic histology, intestinal microbiota, immune response, and resistance against Vibrio alginolyticus and Vibrio parahaemolyticus in whiteleg shrimp (Penaeus vannamei). Six diets were formulated: three diets without L. acidophilus containining 0 (control, ME0), 2.5 (ME2.5), and 5.0 g/kg of moringa (ME5.0) and the same three diets containing L. acidophilus at 1 × 107 CFU/g of diet (ME0+P, ME2.5 + P, and ME5.0 + P, respectively). Growth performance was measured after 60 days of the rearing period. On the final day, the shrimp were sampled to assess enzyme activity, intestinal and hepatopancreatic histology, and gut microbiota. Shrimp hemocytes were examined on Days 0, 1, 2, 4, 7, 14, 21, and 28 to measure the immune response in terms of the total hemocyte count, phenoloxidase activity, phagocytosis, and superoxide anion production. Furthermore, the shrimp were challenged with V. alginolyticus and V. parahaemolyticus. The results revealed that ME2.5 + P significantly increased (P < 0.05) final weight, weight gain, specific growth rate, enzyme activities, and villi height compared with ME2.5 and control. Wall thickness was increased in the shrimp fed diet supplemented with moringa and L. acidophilus compared with the control shrimp. Hepatopancreatic histology revealed that R cells were more abundant in the shrimp fed diet containing moringa and L. acidophilus compared with those fed diet containing moringa alone (P < 0.05) at the same concentration. High-throughput sequencing analysis indicated that the dietary supplementation with moringa and L. acidophilus affected the gut microbiota composition. All gene functions, members of KEGG level 2, related to metabolism were increased in diet supplemented with moringa with or without L. acidophilus compared with the control group. The immune assay revealed that the total hemocyte count, phenoloxidase activity, phagocytic rate, superoxide anion production, and immune-related gene expression (including those of prophenoloxidase II, alpha-2-macroglobulin, penaeidin2, antilipopolysaccharide factor, crustin, lysozyme, glutathione peroxidase, and superoxide dismutase) were higher in the experimental groups than in the control group on several observed days; however, the increases were observed more often in the ME2.5 + P group than in the other treatment groups. Furthermore, the ME2.5 + P group exhibited a significantly higher survival rate (P < 0.05) in the challenge test against V. alginolyticus and V. parahaemolyticus. In conclusion, supplementation with dietary moringa and L. acidophilus at ME2.5 + P improved growth performance, immune system, and resistance against Vibrio in the shrimp.

Abidin Z ，Huang HT ，Hu YF ，Chang JJ ，Huang CY ，Wu YS ，Nan FH ... -  《-》

Dietary SYNSEA probiotic improves the growth of white shrimp, Litopenaeus vannamei and reduces the risk of Vibrio infection via improving immunity and intestinal microbiota of shrimp.

The growth performance, immunological status, and intestinal microbiology of white shrimp, Litopenaeus vannamei, were evaluated after dietary administration of the commercial probiotic SYNSEA. Shrimp were fed a control diet (without probiotic supplement) and two levels of SYNSEA probiotic, a low concentration of SYNSEA (LSL) containing 105 CFU (g diet)-1Bacillus subtilis and 105 CFU (g diet)-1 lactic acid bacteria (LAB), and a high concentration of SYNSEA (LSH) containing 106 CFU (g diet)-1B. subtilis and 106 CFU (g diet)-1 LAB, for 12 weeks. Shrimp fed with the LSL diet significantly increased growth performance as well as final weight and feed efficiency compared to the control, but not the LSH diet. After being orally challenged with Vibrio parahaemolyticus, shrimp fed with LSL diet prior to the challenge or fed with LSL and pathogen simultaneously showed significantly lower mortality compared to the control. SYNSEA probiotic significantly improved shrimp immune response, including lysozyme activity in LSL and LSH groups, and phagocytic activity in the LSL group in comparison to the control. In addition, the gene expressions of anti-lipopolysaccharide factor 2 in LSL and LSH groups, and penaeidin 4 in LSL were also up-regulated. Although there was no significant difference among groups for hepatopancreas and intestinal morphology, the muscular layer thickness and villi height were slightly improved in the intestines of shrimp fed SYNSEA. The 16S rDNA gene amplicon sequence analysis using next-generation sequencing revealed a significant decrease in α-diversity (Margalef's species richness) after oral administration of SYNSEA due to an increase in the relative abundance of beneficial bacteria in the gut flora of shrimp, such as Lactobacillus, Shewanella, and Bradymonadales and a decrease in harmful bacteria, such as Vibrio, Candidatus_Berkiella, and Acinetobacter baumannii. Together the data suggest that the provision of SYNSEA probiotic at 105 CFU (g diet)-1B. subtilis and 105 CFU (g diet)-1 LAB can improve shrimp growth, enhance immunity, and disease resistance status of the host. In addition, these findings conclude that SYNSEA probiotic has great preventive and therapeutic potential for Vibrio infection in shrimp aquaculture.

Lee JW ，Chiu ST ，Wang ST ，Liao YC ，Chang HT ，Ballantyne R ，Lin JS ，Liu CH ... -  《-》

Dietary supplementation with Pediococcus pentosaceus enhances the innate immune response in and promotes growth of Litopenaeus vannamei shrimp.

To reach the sustainable development goals on health management in Litopenaeus vannamei shrimp culture, Pediococcus pentosaceus AB01 was supplemented in shrimp diet. In this study, the control diet and three experimental diets containing P. pentosaceus AB01 (108 , 109 , 1010  CFU/g) were separately introduced to L. vannamei for a 28 days feeding trial. After the feeding trial, percent weight gain, feeding efficiency, and feed conversion ratio (FCR) were significantly elevated in L. vannamei administered with P. pentosaceus AB01 at 109 and 1010 colony-forming unit (CFU)/g. Protease, amylase, and trypsin were found at higher levels in the probiotic-supplied groups. The feeding of shrimps with P. pentosaceus AB01 significantly increased innate immune response and levels of related biochemical parameters in the haemolymph. After the white spot syndrome virus (WSSV) challenge, supplementation of P. pentosaceus AB01 had significant positive effects (p < .05) on survival rate, compared to that of the control diet. The higher resistance of L. vannamei to WSSV might have been due to alterations in the gut microbiome composition and upregulation of the Toll-Like Receptor (TLR) signalling pathway. Hence, P. pentosaceus AB01 may be a promising alternative feed to promote growth rate, modulate microbiota composition, and enhance immunity in L. vannamei shrimp.

Le B ，Kim DG ，Phuoc NN ，Linh NTH ，Yang SH ... -  《-》