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Abstract

The present study aimed to evaluate the individual and combined effects of Lactobacillus rhamnosus (LR) JCM1136 and Lactococcus lactis subsp. lactis (LL) JCM5805 on the growth, intestinal microbiota, intestinal morphology, immune response and disease resistance of juvenile Nile tilapia (Oreochromis niloticus). A total of 720 apparently healthy juvenile Nile tilapia (0.20 ± 0.05 g) were randomly divided into four equal groups. Fish were fed with a basal diet (CK) supplemented with JCM1136 (LR), JCM5805 (LL), and JCM1136 + JCM5805 (LR+LL) at 1 × 108 CFU/g basal diet for 6 weeks, followed by a basal diet for 1 week. After 6 weeks of feeding, the LL treatment significantly increased the growth and feed utilization of Nile tilapia when compared with the CK. Light microscopy and transmission electron microscopy images of the midgut revealed that probiotic supplementation significantly increased gut microvilli length and microvilli density compared to CK. The transcript levels of several key immune-related genes in the mid-intestine and liver of fish were analyzed by means of quantitative polymerase chain reaction (qPCR) at the end of the sixth week. The results showed the following: when compared to CK group, fish in LR had significantly increased transcript levels of IFN-γ, lyzc, hsp70 and IL-1β in the intestine; LL fish showed significantly increased expressions of TNF-α, IFN-γ, lyzc, hsp70 and IL-1β in the intestine and liver; and intestine lyzc, hsp70 and IL-1β and liver TNF-α, IFN-γ, hsp70 and IL-1β were significantly increased in LR+LL fish. Following a 6-week period of being fed probiotics or a control diet, the tilapia were challenged with an intraperitoneal injection of 20 μl of the pathogenic Streptococcus agalactiae (WC1535) (1 × 105 CFU/ml). The survival rates of the probiotic-fed groups were significantly higher than that of the CK group, and the LL group had the highest survival rate. High-throughput sequencing revealed a significantly higher presence of JCM5805 in the guts of LL fish during the period of probiotic application, but this was no longer detected in all LL samples 1 week post cessation of probiotic administration. Cessation of probiotic administration led to disorders of individual gut microbes within the LR and LL groups. Statistical analysis (LEfSe) demonstrated that three phyla, namely, Bacteroidetes, Fusobacteria and Actinobacteria were enriched in the CK group, while the abundance of Proteobacteria was greater in the probiotic-fed fish. At the genus level, Plesiomonas, which includes potential pathogens of fish, were significantly decreased in the probiotic-fed groups. In contrast, a significant increase of Rhizobium and Achromobacter, which can produce a variety of enzymes with cellulolytic and pectolytic activity, were observed in fish fed with probiotics, indicating that dietary probiotics were helpful in the propagation of some probiotic bacteria. Our data revealed that JCM1136 and JCM5805, as a feed additive at 108 CFU/g feed, could improve intestinal morphology, enhance immune status and disease resistance, and affect the gut microbiota of tilapia; thus, these additives could be used as probiotics for juvenile Nile tilapia. JCM5805 was more effective than JCM1136 or the mixture of the two for promoting the growth, enhancing the immune status and disease resistance of tilapia.

Keywords: Immunity; Intestinal microbiota; Lactobacillus rhamnosus JCM1136; Lactococcus lactis JCM5805; Nile tilapia.

Research Insights

SupplementHealth OutcomeEffect TypeEffect Size
Lacticaseibacillus rhamnosusEnhanced Disease ResistanceBeneficial
Large
Lacticaseibacillus rhamnosusImproved Immune ResponseBeneficial
Large
Lacticaseibacillus rhamnosusImproved Intestinal MorphologyBeneficial
Moderate
Lacticaseibacillus rhamnosus R0011Improved Gut MorphologyBeneficial
Moderate
Lacticaseibacillus rhamnosus R0011Improved Immune ResponseBeneficial
Large
Lactobacillus rhamnosusEnhanced Disease ResistanceBeneficial
Large
Lactobacillus rhamnosusImproved Gut MicrobiotaBeneficial
Moderate
Lactobacillus rhamnosusImproved Immune ResponseBeneficial
Large
Lactobacillus rhamnosusImproved Intestinal MorphologyBeneficial
Moderate
Lactobacillus rhamnosusReduced Potential PathogensBeneficial
Moderate
Lactobacillus rhamnosus HA-111Altered Gut Microbiota CompositionBeneficial
Moderate
Lactobacillus rhamnosus HA-111Enhanced Disease ResistanceBeneficial
Large
Lactobacillus rhamnosus HA-111Improved Immune ResponseBeneficial
Large
Lactobacillus rhamnosus HA-111Improved Intestinal MorphologyBeneficial
Moderate
Lactobacillus rhamnosus HA-500Altered Gut MicrobiotaBeneficial
Moderate
Lactobacillus rhamnosus HA-500Enhanced Disease ResistanceBeneficial
Large
Lactobacillus rhamnosus HA-500Improved Immune ResponseBeneficial
Large
Lactobacillus rhamnosus HA-500Improved Intestinal MorphologyBeneficial
Moderate
Lactobacillus rhamnosus IMC 501Enhanced Disease ResistanceBeneficial
Large
Lactobacillus rhamnosus IMC 501Enhanced Gut Probiotic Bacteria PropagationBeneficial
Large
Lactobacillus rhamnosus IMC 501Improved Immune ResponseBeneficial
Large
Lactobacillus rhamnosus IMC 501Improved Intestinal MorphologyBeneficial
Moderate
Lactobacillus rhamnosus IMC 501Reduced Pathogenic Bacteria in GutBeneficial
Large
Lactobacillus rhamnosus LR05Enhanced Disease ResistanceBeneficial
Large
Lactobacillus rhamnosus LR05Enhanced Immune-Related Gene ExpressionBeneficial
Moderate
Lactobacillus rhamnosus LR05Improved Gut MorphologyBeneficial
Moderate
Lactobacillus rhamnosus LR05Reduced Potential PathogensBeneficial
Moderate
Lactobacillus rhamnosus LR06Altered Gut Microbiota CompositionBeneficial
Moderate
Lactobacillus rhamnosus LR06Enhanced Disease ResistanceBeneficial
Large
Lactobacillus rhamnosus LR06Improved Immune ResponseBeneficial
Large
Lactobacillus rhamnosus LR06Reduced Pathogenic Bacteria LevelsBeneficial
Moderate
Lactobacillus rhamnosus Lr-32Enhanced Disease ResistanceBeneficial
Large
Lactobacillus rhamnosus Lr-32Improved Immune ResponseBeneficial
Large
Lactobacillus rhamnosus Lr-32Improved Intestinal MorphologyBeneficial
Moderate
Lactobacillus rhamnosus Lr-32Increased Gut Microvilli LengthBeneficial
Moderate
Lactobacillus rhamnosus Lr-32Reduced Potential Pathogenic BacteriaBeneficial
Moderate
Lactobacillus rhamnosus lysate powderAltered Gut Microbiota CompositionBeneficial
Moderate
Lactobacillus rhamnosus lysate powderEnhanced Disease ResistanceBeneficial
Large
Lactobacillus rhamnosus lysate powderEnhanced Immune-Related Gene ExpressionBeneficial
Moderate
Lactococcus lactis LL-23Enhanced Disease ResistanceBeneficial
Large
Lactococcus lactis LL-23Improved Fish GrowthBeneficial
Large
Lactococcus lactis LL-23Improved Gut MorphologyBeneficial
Moderate
Lactococcus lactis LL-23Improved Immune ResponseBeneficial
Large
Lactococcus lactis subsp. lactis R1058Enhanced Immune Gene ExpressionBeneficial
Large
Lactococcus lactis subsp. lactis R1058Improved Growth RateBeneficial
Large
Lactococcus lactis subsp. lactis R1058Increased Survival Against PathogensBeneficial
Large
Lactococcus lactis subsp. lactis R1058Reduced Potential PathogensBeneficial
Moderate

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