The aim of this study was to examine the effects of dietary protein and lipid levels on the growth performance and homeostasis of the intestinal flora in Paramisgurnus dabryanus. An 8-week 3 × 3 twofactorial experiment was conducted to investigate the interaction between dietary crude protein (CP: 30%, 35%, 40%) and ether extract (EE: 6%, 10%, 14%) on the growth rate and the intestinal microflora of P. dabryanus. A total of 2,160 fish (5.19 ± 0.01 g) were randomly allotted to 36 aquariums each with 60 fish. Fish were fed the experimental diet twice daily. Results revealed that weight gain rate (WGR), specific growth rate (SGR), protein efficiency ratio and net protein utilization significantly increased when increasing protein levels from 30% to 40% (P < 0.05). BothWGR and SGR enhanced first but reduced thereafter with maximum value at 10% lipid level as dietary lipid increased from 6% to 14% (P < 0.05). Significant interactions between protein and lipid were found with feed conversion rate, lipid efficiency ratio and net lipid utilization (P < 0.05). At the phylum level, Proteobacteria and Actinobacteria were the dominant bacteria; at the genus level, Burkholderia-Caballeronia-Paraburkholderia was the dominant bacteria. Fish fed the diet containing 10% lipid had a higher abundance of Proteobacteria and unclassified_f_Eenterobacteriaceae than those fed the 14% lipid diet, and a higher abundance of Rhodobacter than those fed the 6% lipid diet (P < 0.05). Analysis of the predicted functions showed that metabolism in the intestine of fish in the CP40EE10 group was more active than that in CP30EE14 group. Polynomial regression analysis found that a diet containing 40.87% protein and 9.88% lipid can be considered optimal for P. dabryanus.

Docosahexaenoic acid (DHA) is a biologically active fatty acid that reduces the accumulation of lipids. However, the molecular mechanism underlying this process, particularly in fish, is not well understood. Recent studies show that endoplasmic reticulum (ER) stress triggers the activation of the unfolded protein response, which has been revealed to play an essential role in lipid metabolism. In this study, we explored the effect of DHA on ER stress and investigated the potential molecular mechanisms underlying DHA-induced adipocyte lipolysis in grass carp (Ctenopharyngodon idella) both in vivo and in vitro. We found that DHA remarkably reduced the triglyceride content, increased the secretion of glycerol, promoted lipolysis in adipocytes and evoked ER stress, whereas inhibiting ER stress using 4-phenyl butyric acid (4-PBA) inhibited the effects of DHA (P < 0.05). These results implied that ER stress potentially participates in DHA-induced adipocyte lipolysis. Additionally, STF-083010, a specific inositol-requiring enzyme 1α (IRE1α)-inhibitor, attenuated the effects of DHA on lipolysis, demonstrating that IRE1α and X-box binding protein 1 potentially participate in DHA-induced lipolysis. DHA also activated the cyclic adenosine monophosphate (cAMP)-dependent protein kinase A (PKA) pathway by increasing the level of cAMP and activating the PKA enzyme (P < 0.05). Nevertheless, H89, a PKA inhibitor, weakened DHA-induced lipolysis by inhibiting the cAMP/PKA signaling pathway. Furthermore, inhibiting ER stress using 4-PBA also inhibited lipolysis and alleviated DHA-induced activation of the cAMP/PKA signaling pathway, suggesting that ER stress may participate in DHA-induced lipolysis through the activation of the cAMP/PKA signaling pathway. Our data illustrate that DHA supplementation can be a promising nutritional strategy for ameliorating lipid accumulation in grass carp. The present study elucidated the molecular mechanism for DHA-induced lipolysis in grass carp adipocytes and emphasized the importance of ER stress and the cAMP/PKA pathway in DHA-induced lipolysis. These results deepen our understanding of ameliorating lipids deposition in freshwater fish by targeting DHA.

The role of insects in animal nutrition has been reconsidered during recent years, paving the way for an increasing market for edible insects. Their protein and amino acid balance make them a promising source of protein for replacing high value proteins. Yellow mealworm, Tenebrio molitor L. (TM; Coleoptera: Tenebrionidae) larvae, have shown positive effects on broiler performance in several research studies and have a strong potential as a sustainable alternative protein source for monogastric animals. This study aimed to assess the effect of replacing various ratios of basal diets with T. molitor larvae on broiler performance as well as on several meat and welfare characteristics. For the study, 120 one-day-old male chicks (Ross 308) were randomly allocated in 3 treatments and 4 replications (10 birds per pen). Birds of the control group (basal diet) were fed with typical commercial maize and soymeal-based rations in mash form. The other 2 groups were treated with the same diet, after replacing 5% and 10% with dried TM larvae, respectively. On d 35 (end of trial), meat samples were collected and analysed. Body weight, feed intake, body weight gain and feed conversion ratio during the periods of 1 to 10 d (starter period), 11 to 24 d (grower period), 25 to 35 d (finisher period) and 1 to 35 d (total period) were assessed. Pododermatitis, diarrhoea, feather score and litter conditions were also assessed during the trial. The results indicated that TM larvae inclusion in the broilers' diet positively affected body weight gain values, as well as the carcass yield, the meat composition and various welfare traits. Additionally, the dietary treatments with TM larvae favourably affected meat composition and colour parameters, whereas there were also some positive effects on lipid and protein oxidation. Saturated fatty acids were decreased by the dietary supplementation whereas the polyunsaturated fatty acids to SFA ratio increased. In general, the study showed that whole TM larvae addition can provide a promising alternative to soybean meal in the diet of broilers, demonstrating a positive impact on growth, welfare and meat characteristics.

Dietary threonine (Thr) deficiency enhances triglyceride (TG) deposition in the liver of Pekin ducks, which injures hepatic function and impairs growth performance. However, the underlying molecular mechanisms remain unclear. In the present study, we investigated the effects of dietary Thr deficiency on the expressions of proteins and phosphoproteins in liver of Pekin ducks, to identify the underlying molecular changes. A total of 300 one-day-old ducklings were divided into 3 groups with 10 replicates of 10 birds. All ducks were fed corn-wheat-peanut meal diets containing 0.46%, 0.71%, and 0.96% Thr, respectively, from 1 to 21 days of age. Growth performance, serum parameters, hepatic TG content, and expression of genes involved in lipid metabolism of Pekin ducks were determined. A Thr deficiency group (Thr-D, 0.46% Thr) and a Thr sufficiency group (Thr-S, 0.71% Thr) were selected for subsequent proteomic and phosphoproteomic analysis. The results showed that Thr-D reduced the growth performance (P < 0.001), and increased the plasma concentrations of cholesterol, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, and hepatic TG (P < 0.05). Thr-D increased gene expression related to fatty acid and TG synthesis (P < 0.05). A total of 176 proteins and 259 phosphosites (containing 198 phosphoproteins) were observed to be differentially expressed as a result of Thr-D. The upregulated proteins were enriched in the pathway related to amino acid metabolism, peroxisome. The downregulated proteins were enriched in linolenic and arachidonic acid metabolism, and the Janus kinasesignal transducer and activator of transcription (JAK-STAT) signaling pathway. The upregulated phosphoproteins were enriched in the pathways related to fatty acid biosynthesis, fructose and mannose metabolism, and glycolysis/gluconeogenesis. Thr-D reduced the phosphorylation of STAT1 at S729 and STAT3 at S728, and expression of STAT5B. In contrast, Thr-D increased non-receptor tyrosine-protein kinase (TYK2) expression and STAT1 phosphorylation at S649. Taken together, dietary Thr-D increased hepatic TG accumulation by upregulating the expression of genes and proteins, and phosphoproteins related to fatty acid and triglyceride synthesis. Furthermore, these processes might be regulated by the JAK-STAT signaling pathway, especially the phosphorylation of STAT1 and STAT3.

The consequences of feeding broiler chickens with reduced protein (RP) diets for gut health and barrier function are not well understood. This study was performed to elucidate the effect of reducing dietary protein and source of protein on gut health and performance parameters. Four experimental diets included 2 control diets with standard protein levels either containing meat and bone meal (CMBM) or an all-vegetable diet (CVEG), a medium RP diet (17.5% in growers and 16.5% in finisher), and a severe RP diet (15.6% in grower and 14.6% in finisher). Off-sex Ross 308 birds were assigned to each of the 4 diets and performance measurements were taken from d 7 to 42 post-hatch. Each diet was replicated 8 times (10 birds per replicate). A challenge study was conducted on additional 96 broilers (24 birds per diet) from d 13 to 21. Half of the birds in each dietary treatment were challenged by dexamethasone (DEX) to induce a leaky gut. Feeding birds with RP diets decreased weight gain (P < 0.0001) and increased feed conversion ratio (P < 0.0001) from d 7 to 42 compared with control diets. There was no difference between CVEG and CMBM control diets for any parameter. The diet containing 15.6% protein increased (P < 0.05) intestinal permeability independent of the DEX challenge. Gene expression of claudin-3 was downregulated (P < 0.05) in birds fed 15.6% protein. There was a significant interaction between diet and DEX (P < 0.05) and both RP diets (17.5% and 15.6%) downregulated claudin-2 expression in DEX-challenged birds. The overall composition of the caecal microbiota was affected in birds fed 15.6% protein having a significantly lower richness of microbiota in both sham and DEX-injected birds. Proteobacteria was the main phylum driving the differences in birds fed 15.6% protein. At the family level, Bifidobacteriaceae, Unclassified Bifidobacteriales, Enterococcaceae, Enterobacteriaceae, and Lachnospiraceae were the main taxa in birds fed 15.6% protein. Despite supplementation of synthetic amino acids, severe reduction of dietary protein compromised performance and intestinal health parameters in broilers, evidenced by differential mRNA expression of tight junction proteins, higher permeability, and changes in caecal microbiota composition.

Four multi-catheterized lactating goats were used in a 4 × 4 Latin square experiment to investigate the responses of amino acid metabolism in portal-drained viscera (PDV), liver, and mammary glands to short-term varying supplies of methionine (Met). During the last 45 h in each experimental period, goats were fasted for 12 h and then abomasally infused with an amino acid (AA) mixture plus glucose for 33 h. Treatments consisted of graded removal of Met from an infused AA mixture to achieve Met content in the infusate of 100% (complete), 60%, 30%, or 0% that in casein. Graded Met removal decreased the production of milk, milk protein, lactose, and fat linearly whilst also decreasing arterial Met concentration linearly (P < 0.05). Meanwhile, net PDV uptake and liver removal of Met decreased linearly (P < 0.05) due to decreased Met affinity of PDV and liver (P < 0.05). Net mammary uptake of Met (P > 0.1) was maintained as Met supply declined. This was achieved through increased mammary affinity (P < 0.05) and increased mammary blood flow (P < 0.05) totally offsetting the negative effect of decreased circulating Met concentration. Graded removal of Met from the infusate linearly decreased mammary uptake-to-milk output ratios of Met (P < 0.05) and tended to decrease essential amino acid (EAA) linearly (0.05 < P < 0.1). Treatments also linearly decreased circulating concentration of prolactin and linearly increased insulin concentration (P < 0.05). In conclusion, results of the present study indicated there were several mechanisms used to mitigate a Met deficiency, including reduced catabolism of Met in PDV, liver, and peripheral tissue (including mammary glands) and a linear increase in mammary blood flow. The observed decreases in milk protein production as Met supply decreased appear to be a result of regulatory events which may have been driven by decreased circulating prolactin, rather than as a result of decreased mammary Met uptake.

In swine production, stress is a common encounter that leads to serious bacterial infection and adverse effects on growth performance. Though antibiotics have been frequently used to control pathogen spread, sustained negative impacts from antibiotics have been found to affect intestinal integrity and the immune system. Multiple nutritional strategies have shown potential to counteract stress and replace antibiotics, including functional amino acids, low protein diet, plant extracts, organic acids, prebiotics, probiotics, minerals and vitamins. These additives relieve the stress response in swine via different mechanisms and signal transduction pathways. Based on the overview of signaling pathways and stress models, this review highlights the potential of nutritional strategies in swine for preventing or treating stress-related health problems. For wider application in the pig industry, the dose ranges measured require for further validation in different physiological contexts and formulations. In the future, microfluid devices and novel stress models are expected to enhance the efficiency of screening for new antistress candidates.

A 12-wk trial was conducted to compare the tolerance of tilapia to high carbohydrate and high lipid diets. Three isonitrogenous and isoenergetic diets, whose carbohydrate and lipid levels were the following: 35.0% and 8% (control), 44.2% and 4% (D1, high carbohydrate), and 25.8% and 12% (D2, high lipid), respectively. Three hundred tilapias (27 ± 0.11 g) were fed the diets for 10 wk (4 replicates per group); 72 fish from the D1 group were continually fed the D1 (D1D1) and 72 fish from the D2 were continually fed the D2 (D2D2) diet for 2 wk (3 replicates each group) to evaluate the tilapia's capacity to tolerate high carbohydrate and high lipid diets, respectively. Another 36 fish from D1 group were continually fed D2 (D1D2) for comparison with D1D1 and D2D2 groups. In phase 1, hepatosomatic index, liver triglycerides (TG), glucose tolerance (GT) and crude protein in the whole body in D1 group were higher than those in D2 group (P < 0.05). During phase 2, D1D1 group had lower feed intake and weight gain, as well as lower serum total protein and albumin than that of D2D2 group (P < 0.05), while its liver glycogen was significantly higher than that in D1D2 and D2D2 groups (P < 0.05). Moreover, serum glucose and GT were higher in D1D1 and D1D2 groups than those in D2D2 group (P < 0.05). By contrast, D2D2 group had significantly higher intraperitoneal fat, subcutaneous adipose tissue (SCAT) and liver TG than those in D1D1 group (P < 0.05). The mRNA expression of brain npy, hepatic nrf2, gst1 and hepatic transcriptomic data showed that immune-related genes (gama, mrc2, mhc2 and cd163), were downregulated in D1D1 group compared to D2D2 and D1D2 groups. Taken together: 1) tilapia have higher tolerance to a high lipid diet than high carbohydrate diet; 2) despite retention of glucose tolerance, the continuous feeding of D1 diet impaired tilapia's appetite, weight gain rate and host immune response; 3) specific distribution of fat in intraperitoneal regions, SCAT and liver may be a risk-avoidance strategy in tilapia in response to a continuous D2 diet.

This study was conducted to evaluate the influence of dietary lysophospholipids combined with 1% dietary fish oil reduction on the growth performance and hepatic lipid metabolism of largemouth bass (Micropterus salmoides). Five isonitrogenous feeds were prepared with lysophospholipids at 0% (fish oil group, FO), 0.05% (L-0.05), 0.1% (L-0.1), 0.15% (L-0.15) and 0.2% (L-0.2), respectively. The dietary lipid was 11% in the FO diet and 10% in the other diets. Largemouth bass were fed for 68 d (initial body weight = 6.04 ± 0.01 g) with 4 replicates per group and 30 fish per replicate. The results showed that the fish fed diet containing 0.1% lysophospholipids had higher digestive enzyme activity and obtained better growth performance compared to the fish fed FO diet (P < 0.05). The feed conversion rate in the L-0.1 group was significantly lower than that in the other groups. Serum total protein and triglyceride contents in L-0.1 group were significantly higher than those in other groups (P < 0.05) and the contents of total cholesterol and low-density lipoprotein cholesterol in L-0.1 group were significantly lower than those in FO group (P < 0.05). The activity and genes expression of hepatic glucolipid metabolizing enzymes in L-0.15 group were significantly increased compared to those in FO group (P < 0.05). Reducing 1% fish oil along with 0.1% lysophospholipids added to the feed could improve the digestion and absorption of nutrients, enhance the activity of liver glycolipid metabolizing enzymes, and thus effectively promote the growth of largemouth bass.

Intestinal health is critically important for the digestion and absorption of nutrients and thus is a key factor in determining performance. Intestinal health issues are very common in high performing poultry lines due to the high feed intake, which puts pressure on the physiology of the digestive system. Excess nutrients which are not digested and absorbed in the small intestine may trigger dysbiosis, i.e. a shift in the microbiota composition in the intestinal tract. Dysbiosis as well as other stressors elicit an inflammatory response and loss of integrity of the tight junctions between the epithelial cells, leading to gut leakage. In this paper, key factors determining intestinal health and the most important nutritional tools which are available to support intestinal health are reviewed.