Host gut-derived Bacillus probiotics supplementation improves growth performance, serum and liver immunity, gut health, and resistive capacity against Vibrio harveyi infection in hybrid grouper(♀Epinephelus fuscoguttatus × ♂Epinephelus lanceolatus)

Several reports have revealed the vital role that probiotics play in fish growth and health.However,few works are available for host gut-derived probiotics on the growth,immunity,and gut microbiota of fish,especially in hybrid grouper (♀Epinephelus fuscoguttatus×♂Epinephelus lanceolatus) due to their isolation difficulty and functional verification.This study aimed at assessing 3 host gut-derived Bacillus species?effects on the growth,immune and antioxidant-biochemical responses,haematological parameters,intestinal morphology,immune-related gene expression,gut microbiota,and disease resistance against Vibrio harveyi in hybrid grouper.A total of 480 hybrid grouper (initial weight=9.03±0.02 g) were randomly allotted into 4 groups,namely,the group fed a basal diet without probiotic inclusion (control,B0),the group fed the basal diet with Bacillus velezensis GPSAK4 (BV),the group fed the basal diet with Bacillus subtilis GPSAK9 (BS),and the group fed the basal diet with Bacillus tequilensis GPSAK2 (BT) strains at 1.0×10^(9)CFU/g.After a 6-week feeding trial,the results revealed significant improvements (P<0.05) in the growth performance,whole fish-body proximate composition,blood haematological parameters,serum,liver,and intestinal biochemical indexes,intestinal morphology,and protection against V.harveyi pathogen in the probiotic-treated groups compared with the untreated.Additionally,the expressions of intestinal tight junction genes (occludin and ZO1),pro-and anti-inflammatory genes,including IL1β,IL6,IL8,TNFa,MyD88,IL10,and TGFβ,were upregulated (P<0.05) after Bacillus species administration.Host gut-derived Bacillus supplementation shaped the gut microbiota by significantly increasing (P<0.05) the relative abundance of Proteobacteria,Bacteroidetes,Actinobacteria (except the BS group),Acidobacteria(except the BT group),Cyanobacteria (except the BV and BT groups),and Verrucomicrobia phyla,as well as known beneficial genera (Romboutsia,Turicibacter,Epulopiscium,Clostridium_sensu_stricto 1 and 13,Lactobacillus,and Bacillus),but significantly decreased (P<0.05) the abundance of Firmicutes,Chloroflexi,and Fusobacteria phyla,and purported pathogenic genera (Staphylococcus and Photobacterium) compared with the control group.Collectively,the results suggest that B.velezensis GPSAK4,B.subtilis GPSAK9(especially this strain),B.tequilensis GPSAK2 dietary supplementation at 1.0×10^(9)CFU/g has positive effects on the intestinal health of hybrid grouper via microbial composition modulation,thus enhancing the assimilation and absorption of nutrients to boost fish growth,immunity,and disease resistance.

Cell-derived extracellular matrix-coated silk fibroin scaffold for cardiogenesis of brown adipose stem cells through modulation of TGF-β pathway

The cell-derived extracellular matrix(ECM)-modified scaffolds have advantages of mimic tissue specificity and been thought to better mimic the native cellular microenvironment in vitro.ECM derived from cardiac fibroblasts(CFs)are considered as key elements that provide a natural cell growth microenvironment and change the fate of cardiomyocytes(CMs).Here,a new hybrid scaffold is designed based on silk fibroin(SF)scaffold and CFs-derived ECM.CFs were seeded on the SF scaffold for 10days culturing and decellularized to produce CFs-derived ECM-coated SF scaffold.The results showed that the cell-derived ECM-modified silk fibroin scaffold material contained collagen,laminin,fibronectin and other ECM components with myocardial-like properties.Further to explore its effects on brown adipose stem cells(BASCs)differentiation into CMs.We found that the CFderived ECM-coated scaffold also increased the expression of CM-specific proteins(e.g.cardiac troponin T and α-actinin)of BASCs.Notably,the b1-integrin-dependent transforming growth factor-β1 signaling pathway was also involved in the regulation of CF-derived ECM by promoting the differentiation of BASCs into CMs.Overall,these findings provide insights into the bionic manufacturing of engineered cardiac tissues(ECTs)and establish a theoretical basis for the construction of ECTs.

Soybeanβ-conglycinin and glycinin reduced growth performance and the intestinal immune defense and altered microbiome in juvenile pearl gentian groupers Epinephelus fuscoguttatus♀×Epinephelus lanceolatus♂

The utilization efficiency of soy protein is affected by its 2 anti-nutritional substancesdthe antigens bconglycinin and glycinin.This study investigated their effects on the growth performance,intestinal immune defense,andmicrobiome in juvenile pearl gentian groupers(Epinephelus fuscoguttatus♀×Epinephelus lanceolatus♂).Three isonitrogenous and isolipidic diets were formulated containing fishmeal supplemented with 70 g/kgβ-conglycinin or 100 g/kg glycinin,or no supplementation(control).Each experimental diet was fed to quadruplicate groups with 30 fish in each tank for 8 weeks.Dietary inclusion of eitherβ-conglycinin or glycinin significantly reduced weight gain and specific growth rates,and cell proliferation of the distal intestine.Histological evaluation of the intestine tract revealed the inflammation signs,characterized by reducing of plica height and width as well as the number of the goblet cells,and widening of the lamina propria.The group fed theβ-conglycinin diet had reduced lysozyme activity,contents of immunoglobulin M and complements 3 and 4.Increased activities of caspase-3 and-9 were observed in the group fed the bconglycinin diet compared to the other 2 groups.In the intestinal microbiota,the relative abundances of the potentially pathogenic genera Photobacterium and Vibrio were significantly higher in the glycinin group than those in others.Therefore,the existence of soybean antigens(β-conglycinin or glycinin)could damage the structural integrity of the intestine,reduce immune defense,reshape the intestinal microbiome and,ultimately,impair growth in fish.

Subdural hemorrhages in acute lymphoblastic leukemia: case report and literature review

Background:Acute lymphoblastic leukemia is a rare hematological malignancy.Pure subdural hemorrhages in a patient with acute lymphoblastic leukemia patient are extremely rare.Case presentation:This case presented acute spontaneous subdural hemorrhage without head trauma at first,and acute lymphoblastic leukemia was diagnosed later.The second time,the patient was admitted with multiple pure subdural hemorrhages in different locations and periods with a history of slight head trauma.Conclusions:Pure subdural hemorrhages can occur in a patient with acute lymphoblastic leukemia.More care would be needed for pure subdural hemorrhages without obvious head trauma,and patients with hematological malignancies should be protected from even mild head trauma.