Fermented bile acids improved growth performance and intestinal health by altering metabolic profiles and intestinal microbiome in Micropterus salmoides.

A type of fermented bile acids (FBAs) has been produced through a biological method, and its effects on growth performance, metabolism, and intestinal microbiota in largemouth bass were investigated. The results demonstrated that incorporating 0.03 %-0.05 % FBAs diet could improve the final weight, weight gain and specific growth rate, and decrease the feed conversion ratio. Dietary FBAs did not significantly affect the levels of high-density lipoprotein, low-density lipoprotein, and triglycerides, but decreased the activities of α-amylase in most groups. Adding FBAs to the diet significantly increased the integrity of the microscopic structure of the intestine, thickened the muscular layer of the intestine, and notably enhanced its intestinal barrier function. The addition of FBAs to the diet increased the diversity of the gut microbiota in largemouth bass. At the phylum level, there was an increase in the abundance of Proteobacteria, Firmicutes, Tenericutes and Cyanobacteria and a significant decrease in Actinobacteria and Bacteroidetes. At the genus level, the relative abundance of beneficial bacteria Mycoplasma in the GN6 group and Coprococcus in the GN4 group significantly increased, while the pathogenic Enhydrobacter was inhibited. Meanwhile, the highest levels of AKP and ACP were observed in the groups treated with 0.03 % FBAs, while the highest levels of TNF-α and IL-10 were detected in the group treated with 0.04 % FBAs. Additionally, the highest levels of IL-1ß, IL-8T, GF-ß, IGF-1, and IFN-γ were noted in the group treated with 0.06 % FBAs. These results suggested that dietary FBAs improved growth performance and intestinal wall health by altering lipid metabolic profiles and intestinal microbiota in largemouth bass.

The role of UhpA in regulating the virulence gene expression in Edwardsiella piscicida.

Edwardsiella piscicida (E. piscicida) is an important fish pathogen. However, the mechanism of Glu6P transport regulatory protein UhpA how to affect the virulence gene expression in E. piscicida is still unclear. The results in this study showed that the metabolism-related gene expression of cysteine synthase (orf 1134) and sulphate transporter (ychM) in the uhpA mutant strain ΔuhpA was 0.76-fold and 0.68-fold lower than the ones in the wild strains (p < .05). The gene expression of ethA and ethB in the ΔuhpA strain was 0.80-fold and 0.72-fold lower than the ones in the wild strains (p < .05). However, the gene expression of fliC and flgN in the ΔuhpA was 1.51-fold and 1.21-fold higher than the ones in the wild strains (p < .05). The gene expression of T3SS (esrB and esrC) and T6SS (evpB and evpC) in the ΔuhpA was 1.27-fold, 1.13-fold, 1.28-fold and 1.23-fold higher than the ones in the wild strains (p < .05). This suggested that the uhpA gene could regulate the key virulence gene expression, and the uhpA gene was associated with the pathogenicity of E. piscicida in fish.

The UhpA mutant of Edwardsiella piscicida enhanced its motility and the colonization in the intestine of tilapia.

Edwardsiella piscicida (E. piscicida) is a significant bacterial pathogen of cultured fish, which infected fish meanly through the intestine. Glucose 6-phosphate (Glu6P) in the intestine is nutritious to the pathogen, Meanwhile, Glu6P was found using as a virulent regulating signal for bacteria. The UhpA, one of the Glu6P transport system regulatory proteins could down-regulate the uhpC/uhpB/uhpA system and decrease its pathogenicity. However, the motility and the colonization of E. piscicida affected by UhpA were still unclear. In this study, the motility and the colonization of E. piscicida were monitored. The result demonstrated that the motility of EIB202 was significantly stronger than that of in ΔuhpA according to fractions 4, 8 and 9. However, the motility of ΔuhpA was significantly stronger than that of EIB202 according to the total number at the whole experiment. Although, there was no difference in the number of bacteria in the posterior intestine of tilapia after infected with E. piscicida EIB202 and ΔuhpA. The number of bacteria in the anterior and the middle intestine of fish infected with ΔuhpA were significantly higher than that of in fish infected with EIB202 at the whole experiment (P < 0.05). Interestingly, both E. piscicida strains colonized in the anterior intestine than that of in the middle and posterior intestines of tilapia. Besides, the gene expression of IL-1ß and TNF-α in the head-kidney of fish infected with ΔuhpA showed significantly higher (p < 0.05) than fish infected with EIB202 during the whole experimental period. Most importantly, the survival rate of E. piscicida EIB202 and ΔuhpA were 57% and 37% respectively. All results indicate that the uhpA gene mutant in E. piscicida could enhance its motility and the colonization in the intestine of tilapia, this illustrates the mechanism of UhpA decreases the pathogenesis of E. piscicida in fish.

Integrated transcriptomic and metabolomic responses in the hepatopancreas of kuruma shrimp (Marsupenaeus japonicus) under cold stress.

In aquatic ecosystems, the temperature of the water is an important ecological factor that modulates aquatic organisms' metabolism, growth, development, and reproduction. In this study, the morphological, transcriptomic, and metabolomic analyses of response of Marsupeneus japonicus to acute cold stress was investigated. The results revealed that low temperature caused profound morphological damage to the hepatopancreas. Transcriptomic responses suggested that energy and primary metabolism, cytoskeleton structure, and apoptosis signaling were altered. The metabolic responses to cold stress included changes of multiple amino acids and unsaturated fatty acids. Combined transcriptomic and metabolomic data indicated that energy metabolism pathways were downregulated in the hepatopancreas under cold stress. However, M. japonicus increased ATP and unsaturated fatty acids production to ameliorate. Moreover, cold stress caused significant attenuation of macrophage apoptosis. This study provides key information to increase our understanding of low-temperature tolerance in shrimp.

Effects of toxic dinoflagellate Alexandrium catenella on sexual maturation and reproductive output in the pacific oyster Crassostrea gigas.

Marine bivalves which feed mainly on microalgae could accumulate toxins during harmful algal blooms, posing a health hazard to humans and other animals through food chains. The Pacific oyster (Crassostrea gigas) is the globally dominant farmed bivalves in the world. In order to benefit sustainable development of the oyster aquaculture, we assessed the effects of an artificial bloom of the toxin-producing dinoflagellate, Alexandrium catenella, upon sexual maturation in C. gigas. Oysters were exposed to A. catenella from April to June in 2012, and compared to a control batch of oysters fed with Isochrysis galbana. During the exposure, clearance, histological observations, biochemical composition as well as embryonic development were measured. The results indicated that A. catenella could be a food source, and inhibited the clearance rate of C. gigas for I. galbana. Significant pathological changes in the form of degeneration in adductor muscles, mantle, ovary and tubules and several inflammatory responses were observed in C. gigas when exposed to harmful microalga. The exposure of A. catenella had negative effects on assimilation, biochemical composition and so as the reproduction. The results of this study demonstrated that toxic microalga can affect "quality'' of eggs and the consequences, in terms of fertility, embryo and larval output.

The role of Nrf2 in mitigating cadmium-induced oxidative stress of Marsupenaeus japonicus.

Nuclear factor-erythroid 2-related factor-2 (Nrf2) is an important modulator of cellular responses against Cd in mammalian cells. However, whether such modulation is conserved in Marsupenaeus japonicas remains unknown.In our study, the shrimps were injected with dsRNA targeting Nrf2 at 4 µg g-1 body weight (b.w.) or sulforaphane (SFN) at 5 µg g-1 b.w., and then were exposed to 40 mg L-1 CdCl2 for 48 h. After Nrf2 knockdown, the Cd content increased, but decreased in the SFN group. This suggested that Nrf2 could promote Cd excretion. A terminal deoxynulceotidyl transferase nick-end-labeling (TUNEL) assay revealed that the Nrf2 knockdown increased the number of apoptotic cells in M. japonicas, while SFN decreased the number of apoptotic cells. After Nrf2 knockdown, the total antioxidant capacity (T-AOC), superoxide dismutase (Sod) activity, and related gene expression decreased significantly, while the malondialdehyde (MDA) content increased remarkably. By contrast, SFN injection alleviated the oxidative stress, as evidenced by increased T-AOC, Sod activity, sod mRNA expression and a reduced MDA content. Similarly, detoxification related enzyme activities (ethoxyresorufin O-deethylase and glutathione-S-transferase (GST)) and their corresponding gene expressions (cyp3a (cytochrome P450 family 3 subfamily A) and gst) were suppressed in the ds-Nrf2 injection group, while they were elevated in the SFN group. In addition, ds-Nrf2 activated mitochondrial apoptotic pathway, as evidenced the mRNA and protein levels of caspase-3, Bcl2 associated X protein (Bax), and p53, while SFN treatment suppressed them. These results displayed that in M. japonicus Cd-induced cellular oxidative damage probably acts via the Nrf2 pathway.