Preparation, characterization, antimicrobial and cytotoxicity studies of copper/zincloaded montmorillonite

Background： A series of modified montmorillonites（Mt） including zinc-loaded Mt（Zn-Mt）, copper-loaded Mt（Cu-Mt）, copper/zinc-loaded Mt with different Cu/Zn ratio（Cu/Zn-Mt-1, Cu/Zn-Mt-2, Cu/Zn-Mt-3） were prepared by an ion-exchange reaction, and characterized using X-ray diffraction（XRD）, fourier transformed infrared spectroscopy（FTIR） and transmission electron microscopy（TEM）. The specific surface areas, antimicrobial activity and cytotoxicity of the modified Mt were investigated.Results： In the modified Mt, hydrated Cu ions and Zn ions were exchanged in the interlayer space of Mt and the particles were irregular shapes. The results showed that Cu/Zn-Mt enhanced antibacterial and antifungal activity compared with Zn-Mt and Cu-Mt possibly due to the synergistic effect between Cu and Zn. Among the Cu/Zn-Mt with different Cu/Zn raitos, Cu/Zn-Mt with a Cu/Zn ratio of 0.98 or 0.51 showed higher antimicrobial activity against gram-negative bacteria（Escherichia coli）, gram-positive bacteria（Staphylococcus aureus）, fungi（Candida albicans）.Moreover, the antimicrobial activity of Cu/Zn-Mt was correlated with its specific surface area. Cytotoxicity studies on IPEC-J2 cell showed a slight cytotoxicity of Cu/Zn-Mt.Conclusions： The current data provide clear evidence that in terms of its antimicrobial activity and relatively low toxicity, the Cu/Zn-Mt holds great promise for applications in animal husbandry.

Maternal helminth infection protects offspring from high-fat-diet-induced obesity through altered microbiota and SCFAs

Helminth-induced Th2 immunity and gut microbiota have been recently shown to be highly effective in modulating metabolic syndromes in animal models.This study aimed to determine whether maternal immunity and microbial factors affect the induction and development of obesity in offspring.Here,Heligomosomoides polygyrus(Hp)-infected or control female C57BL/6J mice mated with normal males and their offspring were fed a high-fat diet(HFD)for 9 weeks after weaning.Our results showed that Hp-induced maternal outcomes during gestation and lactation significantly impacted offspring metabolic phenotypes.This was evidenced by results showing that offspring from helminth-infected mothers on an HFD(Hp-offspring+HFD)gained significantly less body weight than those from uninfected mothers(Cont-offspring+HFD).Hp-offspring+HFD exhibited no Th2 phenotype but displayed a pattern of gut microbiota composition similar to that of Hp-infected mothers.Cross-fostering experiments confirmed that the helminth-induced maternal attenuation of offspring obesity was mediated through both prenatal and postnatal effects.Our results further showed that helminth-infected dams and their offspring had a markedly altered gut microbiome composition,with increased production of short-chain fatty acids(SCFAs).Intriguingly,Hp-infected mothers and Hp-offspring+HFD showed increased SCFA receptor(GPR)expression in adipose and colonic tissues compared to noninfected mothers and Cont-offspring+HFD,respectively.Moreover,SCFA supplementation to the pups of uninfected control mothers during lactation protected against HFD-induced weight gain,which corresponded with changes in gut bacterial colonization.Collectively,our findings provide new insights into the complex interaction of maternal immune status and gut microbiome,Hp infection,and the immunity and gut microbiome in obese-prone offspring in infant life.

Dietary choline activates the Ampk/Srebp signaling pathway and decreases lipid levels in Pacific white shrimp(Litopenaeus vannamei)

An 8-week feeding trial was conducted in Pacific white shrimp(Litopenaeus vannamei)to evaluate the effects of dietary choline supplementation on choline transport and metabolism,hepatopancreas histological structure and fatty acid profile,and regulation of lipid metabolism.Six isonitrogenous and isolipidic diets were formulated to contain different choline levels of 2.91(basal diet),3.85,4.67,6.55,10.70 and 18.90 g/kg,respectively.A total of 960 shrimp(initial weight,1.38±0.01 g)were distributed randomly into twenty-four 250-L cylindrical fiber-glass tanks,with each diet assigned randomly to 4replicate tanks.The results indicated that dietary choline significantly promoted the deposition of choline,betaine and carnitine(P<0.05).The diameters and areas of R cells,total lipid and triglyceride contents in hepatopancreas,and triglyceride and non-esterified fatty acid contents in hemolymph were negatively correlated with dietary choline level.The contents of functional fatty acids in hepatopancreas,the activity of acetyl-CoA carboxylase(Acc),and the mRNA expression of fas,srebp and acc were highest in shrimp fed the diet containing 4.67 g/kg choline,and significantly higher than those fed the diet containing 2.91 g/kg,the lowest level of choline(P<0.05).The number of R cells,content of very lowdensity lipoprotein(VLDL),activities of carnitine palmitoyl-transferase(Cpt1),lipoprotein lipase and hepatic lipase,and the mRNA expression levels of cpt1,fabp,fatp,ldlr,and ampk in hepatopancreas increased significantly as dietary choline increased(P<0.05).In addition,hepatopancreas m RNA expression levels of ctl1,ctl2,oct1,badh,bhmt,ck,cept,and cct were generally up-regulated as dietary choline level increased(P<0.01).In conclusion,dietary choline promoted the deposition of choline and its metabolites by up-regulating genes related to choline transport and metabolism.Moreover,appropriate dietary choline level promoted the development of hepatopancreas R cells and maintained the normal accumulation of lipids required for development,while high dietary choline not only promoted hepatopancreas lipid export by enhancing VLDL synthesis,but also promoted fatty acidβ-oxidation and inhibited de novo fatty acid synthesis by activating the Ampk/Srebp signaling pathway.These findings provided further insight and understanding of the mechanisms by which dietary choline regulated lipid metabolism in L.vannamei.

High dietary lipid level promotes low salinity adaptation in the marine euryhaline crab(Scylla paramamosain)

The physiological processes involved in adaptation to osmotic pressure in euryhaline crustaceans are highly energy demanding,but the effects of dietary lipids(fat)on low salinity adaptations have not been well evaluated.In the present study,a total of 120 mud crabs(Scylla paramamosain,BW=17.87±1.49 g)were fed control and high-fat(HF)diets,at both medium salinity(23‰)and low salinity(4‰)for 6 wk,and each treatment had 3 replicates with each replicate containing 10 crabs.The results indicated that a HF diet significantly mitigated the reduction in survival rate,percent weight gain and feed efficiency induced by low salinity(P<0.05).Low salinity lowered lipogenesis and activated lipolysis resulting in lipid depletion in the hepatopancreas of mud crabs(P<0.05).Thus,HF diets enhanced the process of lipolysis to supply more energy.In the gills,low salinity and the HF diet increased the levels of mitochondrial biogenesis markers,the activity of mitochondrial complexes,and the expression levels of genes related to energy metabolism(P<0.05).Consequently,the positive effects of the HF diet on energy metabolism in mud crabs at low salinity promoted osmotic pressure regulation.Specifically,significantly higher haemolymph osmotic pressure and inorganic ion content,as well as higher osmotic pressure regulatory enzyme activity in gills,and gene and protein expression levels of NaK-ATPase were observed in crabs fed the HF diet at low salinity(P<0.05).In summary,high dietary lipid levels improved energy provision to facilitate mitochondrial biogenesis,which increased ATP provision for osmotic pressure regulation of mud crabs.This study also illustrates the importance of dietary lipid nutrition supplementation for low salinity adaptations in mud crabs.

Dietary cholesterol promotes growth and ecdysone signalling pathway by modulating cholesterol transport in swimming crabs(Portunus trituberculatus)

Cholesterol,as an indispensable nutrient,regulates molting and growth in crustacean.As crustaceans are unable to biosynthesize cholesterol de novo,it is central to understand how dietary cholesterol affects molting in crustaceans.An 8-week feeding trial was conducted to evaluate the effects of dietary cholesterol level(0.12%,0.43%,0.79%,1.00%,1.30%and 2.50%)on growth,cholesterol metabolism and expression of genes related to lipid and ecdysone metabolism in female swimming crabs(Portunus trituberculatus).A total of 192 crabs(1.41±0.05 g)were randomly distributed into 192 aquaria.Each treatment had 4 replicates with each replicate containing 8 crabs.Crabs fed the 1.00%cholesterol diet showed best growth performance,and thus based on percent weight gain,the optimal dietary cholesterol requirement was calculated at 1.01%.Tissue cholesterol concentrations were positively correlated with dietary cholesterol level.The contents of functional fatty acids in hepatopancreas significantly increased as dietary cholesterol increased from 0.12%to 2.50%(P<0.05).The expression levels of genes related to lipogenesis pathway,lipid catabolism and fatty acid oxidation were significantly downregulated with increased dietary cholesterol level(P<0.05).The highest expression levels of cholesterol transport genes,low-density lipoprotein receptor(ldlr)and low-density lipoprotein receptorrelated protein 2(lrp2)occurred in crabs fed the 1.30%cholesterol diet.Moreover,hormones related to molting such as crustacean hyperglycemic hormone(CHH),methyl farnesoate(MF),molt-inhibiting hormone(MIH),and ecdysone in hemolymph were significantly influenced by dietary cholesterol level(P<0.05).The highest expression levels of ecdysone receptor(ecr)and chitinase 1(chi1)in eyestalk and hepatopancreas were found in crabs fed the diet containing 1.00%cholesterol(P<0.05).In conclusion,the optimal dietary level was beneficial to functional fatty acid accumulation,regulated lipid metabolism,promoted the ecdysone signalling pathway by improving the cholesterol transport,and improved the molting rate and growth of swimming crabs.