Dietary chenodeoxycholic acid attenuates high-fat diet-induced growth retardation, lipid accumulation and bile acid metabolism disorder in the liver of yellow catfish Pelteobagrus fulvidraco.

This experiment was conducted to investigate whether dietary chenodeoxycholic acid (CDCA) could attenuate high-fat (HF) diet-induced growth retardation, lipid accumulation and bile acid (BA) metabolism disorder in the liver of yellow catfish Pelteobagrus fulvidraco. Yellow catfish (initial weight: 4·40 (sem 0·08) g) were fed four diets: the control (105·8 g/kg lipid), HF diet (HF group, 159·6 g/kg lipid), the control supplemented with 0·9 g/kg CDCA (CDCA group) and HF diet supplemented with 0·9 g/kg CDCA (HF + CDCA group). CDCA supplemented in the HF diet significantly improved growth performance and feed utilisation of yellow catfish (P < 0·05). CDCA alleviated HF-induced increment of hepatic lipid and cholesterol contents by down-regulating the expressions of lipogenesis-related genes and proteins and up-regulating the expressions of lipololysis-related genes and proteins. Compared with the control group, CDCA group significantly reduced cholesterol level (P < 0·05). CDCA significantly inhibited BA biosynthesis and changed BA profile by activating farnesoid X receptor (P < 0·05). The contents of CDCA, taurochenodeoxycholic acid and glycochenodeoxycholic acid were significantly increased with the supplementation of CDCA (P < 0·05). HF-induced elevation of cholic acid content was significantly attenuated by the supplementation of CDCA (P < 0·05). Supplementation of CDCA in the control and HF groups could improve the liver antioxidant capacity. This study proved that CDCA could improve growth retardation, lipid accumulation and BA metabolism disorder induced by HF diet, which provided new insight into understanding the physiological functions of BA in fish.

Klf4-Sirt3/Pparα-Lcad pathway contributes to high phosphate-induced lipid degradation.

BACKGROUND: Phosphorus commonly reduces lipid deposition in the vertebrates. However, the underlying mechanisms involved in the process remain unclear. METHODS: Yellow catfish were given three experimental diets with dietary phosphate levels of 3.22, 6.47 and 7.99 g Pi kg- 1, respectively, for 8 weeks. The contents of triglyceride, non-esterified free fatty acids, adenosine triphosphate, nicotinamide adenine dinucleotide, nicotinamide adenine dinucleotide, enzymatic activities, mRNA and protein expression were determined in the intestinal tissues. Hematoxylin and eosin, Oil Red O staining, and transmission electron microscope were performed for intestinal tissues. Primary intestinal epithelial cells were isolated from yellow catfish intestine. Western blot analysis, Immunoprecipitation assays, Immunofluorescence staining, and RNA extraction and quantitative real-time PCR were decided. Luciferase reporter assays and electrophoretic mobility shift assay were used to evaluate the function of Sirt3, PPARα and Lcad promoters. RESULTS: High dietary phosphate intake activated intestinal phosphate absorption and excretion, and reduced lipid deposition through increasing lipolysis in the intestine. Moreover, phosphate incubation increased the mRNA and protein expression of krüppel like factor 4 (klf4), silent mating-type information regulation 2 homolog 3 (sirt3), peroxisome proliferator activated receptor alpha (pparα) and long chain acyl-CoA dehydrogenase (lcad) in the intestinal epithelial cells (IECs), and klf4 knockdown attenuated the phosphate-induced increase of protein levels of Sirt3, Pparα and Lcad. Further investigation found that Klf4 overexpression increased the activity of sirt3 and pparα promoters, which in turn reduced the acetylation and protein level of Lcad. CONCLUSION: Dietary Pi excess induced lipid degradation by the activation of the Klf4-Sirt3/Pparα-Lcad pathway in the intestine and primary IECs. Video Abstract.

Genome reanalysis to decipher resistome, virulome, and attenuated characters of attenuated Streptococcus agalactiae strain HZAUSC001.

Streptococcus agalactiae is a serious pathogen causing severe anthropozoonosis in a broad range of hosts, from aquatic animals to mammals, including humans. S. agalactiae HZAUSC001 was isolated from a moribund tilapia fish exhibiting classic clinical symptoms of streptococcosis in Zhanjiang, Guangdong, China. And it was identified as the etiological factor resulting in fish disease, but was notable because it exhibited attenuated virulence. Here, the genome of S. agalactiae HZAUSC001 was re-analyzed; we assessed the resistome and virulome and deciphered the attenuated characters of HZAUSC001. The S. agalactiae HZAUSC001 genome was assembled into one chromosome with a GC-content of 35.37% and 1972 predicted open reading frames (ORFs). Phylogenetic analysis indicated that it is evolutionarily similar to piscine GBS strains GD201008-001 and ZQ0910. After re-analyzing the published genomic sequence of HZAUSC001, we identified 38 virulence factor genes and one antibiotic-resistance gene. Note that three previously noted virulence genes, bca (C protein alpha-antigen), cpbA (choline-binding protein A) and esp (enterococcal surface protein), were absent in the virulence-attenuated strain S. agalactiae HZAUSC001 but present in the highly virulent strain S. agalactiae GD201008-001. We speculate that the absence of these three virulence genes may be associated with the attenuated traits of the HZAUSC001 strain. Collectively, our study supports that HZAUSC001 may be an excellent candidate for development of an attenuated vaccine, and our results contribute to further understanding of GBS epidemiology and surveillance targets.

Comparative transcriptome profiling reveals a mechanism of Streptococcus agalactiae resistance to florfenicol.

Florfenicol is widely used to control diseases in aquatic animals, and is used extensively to treat streptococcosis-caused by Streptococcus agalactiae-in the commercially important fish tilapia. There are known issues with the development of florfenicol resistance in Streptococcus agalactiae, but the underlying resistance mechanisms are not clear, a situation currently preventing optimal deployment of antibiotics. Here, we examined the induction of resistance by successively increasing the concentrations of florfenicol, and then used RNA-sequencing (RNA-Seq) to characterize changes in the transcriptomes of a florfenicol-resistant strain (H51-R) and a florfenicol-sensitive strain (H51-S). We obtained a total of 18,418,068 sequence reads in H51-R and 16,070,122 sequence reads in H51-S, from which a total of 1940 unigenes were assembled. In total, 376 unigenes were found to be differently expressed genes (DEGs). After florfenicol treatment, 181 genes were upregulated and 195 genes were downregulated. GO functional analysis of the DEGs indicated that the most strongly enriched GO terms included metabolic process (152 genes), catalytic activity (146), and binding (133), with terms including membrane, membrane part, and transporter activity also showing enrichment. KEGG pathway enrichment analysis highlighted that ribosomes were prominently involved in the transcriptional changes associated with florfenicol resistance. This study demonstrates that florfenicol treatment affects multiple biological functions of Streptococcus agalactiae, suggests that florfenicol resistance in Streptococcus agalactiae is closely related to the reduction of intracellular drug accumulation caused by ATP-binding cassette (ABC) transporters, and highlights the potential involvement of altered ribosomal function in florfenicol resistance.

The pathogenic and antimicrobial characteristics of an emerging Streptococcus agalactiae serotype IX in Tilapia.

Streptococcus agalactiae (S. agalactiae, GBS) infection has caused significant economic loss in the tilapia aquaculture, which is one of the most important commercial fish worldwide. Among the 10 serotypes of GBS, serotypes Ia, Ib, II and III were epidemic in tilapia while serotype IX has never been found in tilapia before. In this study, 80 strains isolated from moribund tilapia in China were identified as GBS. All the isolates have been classified as serotype III or serotype IX of GBS. Unexpectedly, the serotype IX has never been reported in fish, but it was epidemic in mammals. Antimicrobial resistance results showed that serotype IX but not III was resistant to streptomycin and erythromycin. Artificial infection results showed that both serotypes could cause serious pathological injuries in the infected tissues of tilapia. Furthermore, serotype IX instead of serotype III, mainly infected the brain of tilapia. The results will shed a new light on the epidemic and pathogenicity of GBS, and will pave a new way for the prevention of Streptococcosis in tilapia.

GapA, a potential vaccine candidate antigen against Streptococcus agalactiae in Nile tilapia (Oreochromis niloticus).

Streptococcosis due to the bacterium Streptococcus agalactiae (S. agalactiae) has resulted in enormous economic losses in aquaculture worldwide, especially in the tilapia culture industry. Previously, there were limited vaccines that could be employed against streptococcosis in tilapia. This study aimed to develop a vaccine candidate using the glyceraldehyde-phosphate dehydrogenase protein (GapA) of S. agalactiae encoded by the gapA gene. Tilapia were intraperitoneally injected with PBS, PBS + Freund's adjuvant, PBS + Montanide's adjuvant, GapA + Freund's adjuvant, GapA + Montanide's adjuvant, killed S. agalactiae whole cells (WC)+Freund's adjuvant, or killed S. agalactiae whole cells (WC)+ Montanide's adjuvant. They were then challenged with S. agalactiae, and the relative percentage survival (RPS) was monitored 14 days after the challenge. The highest RPSs were observed in the WC groups, with 76.7% in WC + Freund's adjuvant and 74.4% in WC + Montanide's adjuvant groups; these were followed by the GapA groups, with 63.3% in GapA + Freund's adjuvant and 45.6% in GapA + Montanide's adjuvant groups. The RPS of the PBS group was 0%, and those of PBS + Freund's adjuvant and PBS + Montanide's adjuvant groups were 6.7% and 3.3%, respectively. Additionally, the IgM antibody responses elicited in GapA groups and WC groups were significantly higher than those in PBS groups. Furthermore, the expressions of cytokine (IL-1ß and TNF-α) mRNAs in the GapA groups and WC groups were significantly higher than those in the PBS groups. Taken together, these results reveal that the GapA protein is a promising vaccine candidate that could be used to prevent streptococcosis in tilapia.

Draft Genome Sequence of an Attenuated Streptococcus agalactiae Strain Isolated from the Gut of a Nile Tilapia (Oreochromis niloticus).

Streptococcus agalactiae is a pathogen that causes severe anthropozoonosis within a broad range of hosts from aquatic animals to mammals, including human beings. Here, we describe the draft genome of S. agalactiae HZAUSC001, a low-virulent strain isolated from the gut of a moribund tilapia (Oreochromis niloticus) in China.