Autophagy: Are Amino Acid Signals Dependent on the mTORC1 Pathway or Independent?

Autophagy is a kind of "self-eating" phenomenon that is ubiquitous in eukaryotic cells. It mainly manifests in the damaged proteins or organelles in the cell being wrapped and transported by the autophagosome to the lysosome for degradation. Many factors cause autophagy in cells, and the mechanism of nutrient-deficiency-induced autophagy has been a research focus. It has been reported that amino-acid-deficiency-induced cellular autophagy is mainly mediated through the mammalian rapamycin target protein complex 1 (mTORC1) signaling pathway. In addition, some researchers also found that non-mTORC1 signaling pathways also regulate autophagy, and the mechanism of autophagy occurrence induced by the deficiency of different amino acids is not precisely the same. Therefore, this review aims to summarize the process of various amino acids regulating cell autophagy and provide a narrative review on the molecular mechanism of amino acids regulating autophagy.

Borrelia miyamotoi Infections in Humans and Ticks, Northeastern China.

We conducted an investigation of Borrelia miyamotoi infections in humans and ticks in northeastern China. Of 984 patients reporting recent tick bites, 14 (1.4%) were found to be infected with B. miyamotoi by PCR and genomic sequencing. The 14 patients had nonspecific febrile manifestations, including fever, headache, anorexia, asthenia, and arthralgia. Rash, eschar, and regional lymphadenopathy were each observed in 1 patient. Four (28.6%) patients were hospitalized because of severe disease. B. miyamotoi was detected in 3.0% (19/627) of Ixodes persulcatus, 1 (2.8%) of 36 Haemaphysalis concinna, and none of 29 Dermacentor silvarum ticks. Phylogenetic analyses based on sequences of a nearly entire 16s rRNA gene, a partial flagellin gene, and the glycerophosphodiester phosphodiesterase gene revealed that B. miyamotoi identified in patients and ticks were clustered in the group of the Siberian type. These findings indicate that B. miyamotoi is endemic in northeastern China and its public health significance deserves further investigation.

Dietary supplementation with anthocyanin attenuates lipopolysaccharide-induced intestinal damage through antioxidant effects in yellow-feathered broiler chicks.

This study investigated the protective effects of anthocyanin (AC) supplementation on lipopolysaccharide (LPS)-challenged yellow-feathered broiler chicks. A total of 480 1-d female broiler chicks were randomly assigned to 4 treatment groups: basal diet (CON), basal diet + LPS-challenge (LPS), supplementation with 100 or 400 mg/kg AC + LPS-challenge (AC100, AC400). On d 17 and d 19, birds in LPS, AC100 and AC400 received an intramuscular dose of LPS, while birds in CON received saline. The result showed that (1) LPS injection significantly decreased (P < 0.05) body weight on d 21 and average daily gain of broiler chicks from 1 to 21 days of age, and supplementation with 100 mg/kg AC increased (P < 0.05) those of LPS-challenged broilers. (2) There were no differences among the treatments (P > 0.05) in relative weights of immune organs. (3) Supplementation with AC (AC100 and AC400) increased (P < 0.05) the jejunal villus height and villus height/crypt depth ratio (AC100) of LPS-challenged birds. Challenge with LPS decreased the relative expression of OCLN (Occludin), ZO-1, JAM2, and MUC2 in jejunal mucosa of broilers, and supplementation with AC offset the relative expression of ZO-1, JAM2 (AC100 and AC400), and OCLN (AC400) in LPS-injected broilers. (4) LPS-induced increase in the malondialdehyde (MDA) concentration and decreases in activity of total superoxide dismutase (T-SOD), and expression of SOD1, CAT and GPX in jejunal mucosa, were attenuated by dietary AC supplementation. In conclusion, in yellow-feathered broiler chicks, dietary supplementation with AC alleviated LPS-induced declined growth performance and mucosal damage of the intestine through antioxidant effects.

Dietary supplementation of bilberry anthocyanin on growth performance, intestinal mucosal barrier and cecal microbes of chickens challenged with Salmonella Typhimurium.

BACKGROUND: Anthocyanins (AC) showed positive effects on improving the intestinal health and alleviating intestinal pathogen infections, therefore, an experiment was conducted to explore the protective effects of supplemented AC on Salmonella-infected chickens. METHODS: A total of 240 hatchling chickens were randomly allocated to 4 treatments, each with 6 replicates. Birds were fed a basal diet supplemented with 0 (CON, and ST), 100 (ACL) and 400 (ACH) mg/kg of AC for d 60, and orally challenged with PBS (CON) or 109 CFU/bird (ST, ACL, ACH) Salmonella Typhimurium at d 14 and 16. RESULTS: (1) Compared with birds in ST, AC supplementation increased the body weight (BW) at d 18 and the average daily gain (ADG) from d 1 to 18 of the Salmonella-infected chickens (P < 0.05); (2) AC decreased the number of Salmonella cells in the liver and spleen, the contents of NO in plasma and inflammatory cytokines in ileal mucosa of Salmonella-infected chickens (P < 0.05); (3) Salmonella infection decreased the ileal villi height, villi height to crypt depth (V/C), and the expression of zonulaoccludins-1 (ZO-1), claudin-1, occludin, and mucin 2 (MUC2) in ileal mucosa. AC supplementation relieved these adverse effects, and decreased ileal crypt depth (P < 0.05); (4) In cecal microbiota of Salmonella-infected chickens, AC increased (P < 0.05) the alpha-diversity (Chao1, Pd, Shannon and Sobs indexes) and the relative abundance of Firmicutes, and decreased (P < 0.05) the relative abundance of Proteobacteria and Bacteroidota and the enrichment of drug antimicrobial resistance, infectious bacterial disease, and immune disease pathways. CONCLUSIONS: Dietary AC protected chicken against Salmonella infection via inhibiting the Salmonella colonization in liver and spleen, suppressing secretion of inflammatory cytokines, up-regulating the expression of ileal barrier-related genes, and ameliorating the composition and function of cecal microbes. Under conditions here used, 100 mg/kg bilberry anthocyanin was recommended.

Dietary berberine supplementation improves growth performance and alleviates gut injury in weaned piglets by modulating ileal microbiota and metabolites.

This study investigated the effects of dietary berberine (BBR) supplementation on the growth performance, intestinal health, and ileal microbiome and metabolomic profile in weaned piglets challenged with enterotoxigenic Escherichia coli (ETEC). Dietary BBR supplementation significantly attenuated the reduced average daily gain (ADG) and attenuated the increased feed to gain ratio (F/G) and the incidence of diarrhea induced by ETEC K88 (P < 0.05). Dietary BBR supplementation significantly increased the villus height and the villus height to crypt depth ratio in the ileum (P < 0.05). Moreover, the mRNA expression of ZO-1 and occludin as well as aquaporins (AQP1, AQP3, AQP4, AQP7, and AQP10) and Na+/H+ exchanger 3 (NHE3) in ileal mucosa was significantly upregulated by BBR treatment (P < 0.05). Additionally, BBR treatment significantly inhibited the increase of interleukin-1ß (IL-1ß) in jejunal mucosa caused by ETEC and reduced the levels of tumor necrosis factor-α (TNF-α) and IL-1ß and increased interleukin-10 (IL-10) in colonic mucosa (P < 0.05). Dietary BBR treatment significantly increased the Observed_species, Chao 1, abundance based coverage estimators (ACE), and PD_whole tree in the ileal digesta of weaned piglets challenged with ETEC. At the genus level, the relative abundance of unidentified Clostridiales was decreased, while Weissella, Alloprevotella, unidentified Prevotellaceae, and Catenibacterium were increased in the BBR + ETEC group when compared to the ETEC group (P < 0.05). Spearman correlation analysis showed that the relative abundance of unidentified Clostridiales (genus) was negatively correlated with the ileal villus height but negatively correlated with diarrhea and intestinal IL-1ß and TNF-α concentrations (P < 0.05). The ileal metabolome analysis showed that the metabolic pathways including primary and secondary bile acid biosynthesis and bile secretion were significantly enriched by BBR treatment. Collectively, dietary BBR supplementation effectively improved the growth performance and alleviated the diarrhea and intestinal injury induced by ETEC K88 in weaned piglets, which might closely involve the modulation of ileal microbiota and metabolites.

Transcriptome and proteome profile of jejunum in chickens challenged with Salmonella Typhimurium revealed the effects of dietary bilberry anthocyanin on immune function.

Introduction: The present study investigated the effects of bilberry anthocyanin (BA) on immune function when alleviating Salmonella Typhimurium (S. Typhimurium) infection in chickens. Methods: A total of 180 newly hatched yellow-feathered male chicks were assigned to three groups (CON, SI, and SI + BA). Birds in CON and SI were fed a basal diet, and those in SI + BA were supplemented with 100 mg/kg BA for 18 days. Birds in SI and SI + BA received 0.5 ml suspension of S. Typhimurium (2 × 109 CFU/ml) by oral gavage at 14 and 16 days of age, and those in CON received equal volumes of sterile PBS. Results: At day 18, (1) dietary BA alleviated weight loss of chickens caused by S. Typhimurium infection (P < 0.01). (2) Supplementation with BA reduced the relative weight of the bursa of Fabricius (P < 0.01) and jejunal villus height (P < 0.05) and increased the number of goblet cells (P < 0.01) and the expression of MUC2 (P < 0.05) in jejunal mucosa, compared with birds in SI. (3) Supplementation with BA decreased (P < 0.05) the concentration of immunoglobulins and cytokines in plasma (IgA, IL-1ß, IL-8, and IFN-ß) and jejunal mucosa (IgG, IgM, sIgA, IL-1ß, IL-6, IL-8, TNF-α, IFN-ß, and IFN-γ) of S. Typhimurium-infected chickens. (4) BA regulated a variety of biological processes, especially the defense response to bacteria and humoral immune response, and suppressed cytokine-cytokine receptor interaction and intestinal immune network for IgA production pathways by downregulating 6 immune-related proteins. Conclusion: In summary, the impaired growth performance and disruption of jejunal morphology caused by S. Typhimurium were alleviated by dietary BA by affecting the expression of immune-related genes and proteins, and signaling pathways are related to immune response associated with immune cytokine receptors and production in jejunum.

Skin infectome of patients with a tick bite history.

Introduction: Ticks are the most important obligate blood-feeding vectors of human pathogens. With the advance of high-throughput sequencing, more and more bacterial community and virome in tick has been reported, which seems to pose a great threat to people. Methods: A total of 14 skin specimens collected from tick-bite patients with mild to severe symptoms were analyzed through meta-transcriptomic sequencings. Results: Four bacteria genera were both detected in the skins and ticks, including Pseudomonas, Acinetobacter, Corynebacterium and Propionibacterium, and three tick-associated viruses, Jingmen tick virus (JMTV), Bole tick virus 4 (BLTV4) and Deer tick mononegavirales-like virus (DTMV) were identified in the skin samples. Except of known pathogens such as pathogenic rickettsia, Coxiella burnetii and JMTV, we suggest Roseomonas cervicalis and BLTV4 as potential new agents amplified in the skins and then disseminated into the blood. As early as 1 day after a tick-bite, these pathogens can transmit to skins and at most four ones can co-infect in skins. Discussion: Advances in sequencing technologies have revealed that the diversity of tick microbiome and virome goes far beyond our previous understanding. This report not only identifies three new potential pathogens in humans but also shows that the skin barrier is vital in preventing horizontal transmissions of tick-associated bacteria or virus communities to the host. It is the first research on patients' skin infectome after a tick bite and demonstrates that more attention should be paid to the cutaneous response to prevent tick-borne illness.

Feed Restriction Improves Lipid Metabolism by Changing the Structure of the Cecal Microbial Community and Enhances the Meat Quality and Flavor of Bearded Chickens.

Excessive fat deposition in full-fed Bearded chickens does not only reduce carcass yield but also causes consumer rejection of meat. Feed restriction (FR) is an effective method to save on feed cost, reduce carcass fat deposition, and improve meat quality. A total of 560 150-d Bearded chickens were randomly divided into seven groups (each with eight replicates of ten birds) for 40 days. The control group was fed with the basal diet ad libitum (CON), and the other six groups were fed with 90% of the feed intake (90% FI), 80% FI, 70% FI, 90% metabolizable energy (90% ME), 80% ME, and 70% ME of the CON, respectively. Compared to the CON group, FR increased meat yield, but the total weight of the Bearded chickens was slighter; 80% FI and 70% ME improved the relative lipid metabolism indices of chickens, especially the levels of triglycerides and total cholesterol in the plasma and liver (p < 0.05), and decreased calpastatin activity in the breast muscle (p < 0.05). Additionally, 16S rRNA sequencing of cecal microbial community indicated that an increase in the abundance of Hydrogenoanaerobacterium and Bacteroides plebeius was observed in the 80% FI group (p < 0.05), and an enrichment in Olsenella, Catabacter, and Lachnospiraceae were observed in the 70% ME group (p < 0.05) compared to the CON group. Moreover, compared to the CON group, the L * value of the breast muscle significantly decreased, and a * value significantly increased in the 80% FI group (p < 0.05). Notably, the concentrations of threonine, lysine, aspartic acid, glutamic acid, proline, and arginine and the activity of calpain in breast muscle increased in the 80% FI group more than in the CON group (p < 0.05), while valine, isoleucine, leucine, phenylalanine, lysine, alanine, tyrosine and proline decreased in ME restriction groups (p < 0.05). Taken together, our results indicated that 80% FI could improve lipid metabolism by changing the structure of the cecal microbial community, and the meat quality and flavor of the Bearded chickens in 80% FI group was improved with a promoted meat color score, flavor substances, and the calproteinase system.

Antrodia cinnamomea polysaccharide improves liver antioxidant, anti-inflammatory capacity, and cecal flora structure of slow-growing broiler breeds challenged with lipopolysaccharide.

Lipopolysaccharides (LPS) induces liver inflammatory response by activating the TLR4/NF-κB signaling pathway. Antrodia cinnamomea polysaccharide (ACP) is a medicinal mushroom that can protect from intoxication, liver injury, and inflammation. Nevertheless, the effect of ACP on the liver antioxidant, anti-inflammatory capacity and cecal flora structure of LPS-challenged broilers remains unclear. The aim of this experiment was to investigate the effects of ACP on the anti-oxidative and anti-inflammatory capacities of the liver, and cecal microbiota in slow-growing broilers stimulated by LPS. A total of 750 slow-growing broilers (9-day-old) were assigned to five treatments with 6 replicates of 25 chicks per replicate: a control diet, the chicks were fed a control diet and challenged with LPS. Dietary treatments 3 to 5 were the control diet supplemented with 100, 200, 400 mg/kg ACP challenged with LPS, respectively. The groups of 100 mg/kg ACP supplementation significantly increased liver index, pancreas index, and bursa of Fabricius index (P < 0.05). The GSH-Px content of LPS-challenged broilers was lower than that of the control group (P < 0.001), but the content of MDA increased (P < 0.001). Feeding with 100 mg/kg ACP resulted in increased the activity of T-AOC, GSH-Px, and T-SOD, and decreased MDA content (P < 0.05). The activity of TNF-α, IL-1ß, and IL-6 of the LPS group increased, but these indicators were decreased with supplemental 100 mg/kg ACP (P < 0.05). Dietary application of ACP up to 100 mg/kg down-regulated (P < 0.05) the expression of TLR4/NF-κB pathway in the liver induced by LPS. The results of 16S rRNA demonstrated that feeding with 100 mg/kg ACP can change the diversity and composition of the gut microbiota, and restrained the decline of beneficial cecal microbiota (typically Lactobacillus, Faecalibacterium, and Christensenellaceae R-7 group) in the challenged LPS group (P < 0.05). Conclusively, feeding a diet with 100 mg/kg ACP may have beneficial effects on liver damage and the bacterial microbiota diversity and composition in the ceca of LPS-stressed slow-growing broiler breeds, probably because of its combined favorable effects on antioxidants and cytokines contents, and restoration the decline of beneficial cecal microbiota.

Arginine Promotes the Expression of Aquaporin-3 and Water Transport in Porcine Trophectoderm Cells Through NO- and cAMP-Dependent Mechanisms.

BACKGROUND: Dietary supplementation with L-arginine (Arg) has been shown to increase the volume of fetal fluids in gestating swine. Aquaporins (AQPs), known as water channel proteins, are essential for embryonic growth and development. It was not known if Arg mediates water transport through AQPs in porcine conceptus trophectoderm (pTr2) cells. METHODS: pTr2 cells derived from pregnant gilts on day 12 of gestation were cultured in customized Arg-free Dulbecco's modified Eagle's Ham medium (DMEM) supplemented with either 0.00, 0.25, or 0.50 mM Arg. RESULTS: Arg treatment increased water transport and the expression of AQP3, which was abundantly expressed in pTr2 cells at both the mRNA and protein levels. Arg also increased the expression of iNOS and the synthesis of nitric oxide (NO) in pTr2 cells. The presence of Nω-nitro-L-arginine methyl ester hydrochloride (L-NAME; an inhibitor of NO synthase) significantly attenuated the Arg-induced expression of AQP3. Furthermore, 0.50 mM Arg increased the concentrations of cAMP and the abundances of phosphorylated cAMP-dependent protein kinase A (PKA), phosphorylated PKA α/ß/γ, and phosphorylated CREB. These effects of Arg were mimicked by Forskolin (a cell-permeable activator of adenylyl cyclase), but inhibited by H-89 (an inhibitor of cAMP-dependent protein kinase). CONCLUSIONS: The results of this study demonstrate that Arg regulates AQP3 expression and promotes water transport in pTr2 cells through NO- and cAMP-dependent signaling pathways.

Optimal Level of Supplemental Manganese for Yellow-Feathered Broilers during the Growth Phase.

This experiment investigated the effect of an optimized supplemental dietary manganese (Mn) on growth performance, tibial characteristics, immune function and meat quality, of yellow-feathered broilers. In three rearing periods, birds were fed for 21-d periods, from d 1 (starter), d 22 (grower) and d 43 (finisher), respectively, with basal diets (containing 16, 17, and 14 mg/kg analyzed Mn, respectively) supplemented with 0, 20, 40, 60, 80, 100, 120 and 140 mg/kg Mn. For starter phase broilers, supplemental manganese affected feed to gain ratio (F/G), and the minimum value was observed with 120 mg/kg manganese. During the grower phase, ADG increased quadratically (p < 0.05) with supplemental Mn and was maximal with 54 mg/kg additional manganese estimated using the regression equation. There was no influence of supplemental manganese on growth performance of broilers during the finisher phase (p > 0.05). The thymic relative weight of broilers were linearly (p < 0.05) and quadratically (p < 0.05) increased with supplemental Mn and maxima were obtained with 95 and 110 mg/kg additional Mn at 42 d and 63 d. The bone density of the tibia in broilers at d 21, 42 and 63 were increased quadratically (p < 0.05) by supplemental Mn, and optimal supplementation for the three phases was 52, 60 and 68 mg/kg, respectively. The weight, diameter, breaking strength and bone density of the tibia of 63-d broilers were influenced (p < 0.05) by supplemental manganese. The lightness (L*) value (linear, p < 0.05) and yellowness (b*) value (p < 0.05) of the breast muscle were decreased by dietary manganese supplementation, and the optimal supplementation, based on L*, was 86 mg/kg. In conclusion, supplemental Mn affected the growth performance, thymic relative weight, tibial characteristics, and the meat color of yellow-feathered broilers. From the quadratic regressions, the optimal supplementation of yellow-feathered broilers at the starter, grower and finisher phases to achieve the best performance was 52, 60, and 68 mg/kg, respectively.

Effects of Dietary Supplementation with Bilberry Extract on Growth Performance, Immune Function, Antioxidant Capacity, and Meat Quality of Yellow-Feathered Chickens.

The experiment was conducted to investigate the effects of bilberry extract on growth performance, meat quality, antioxidant status, and immune function of yellow-feathered chickens. A total of 360 female hatchling Lingnan chickens were randomly allocated to three treatments with 6 replicates of 20 chickens per replicate. Birds were fed a basal diet supplemented with 0 (the control group), 100 (B100), and 400 (B400) mg/kg of bilberry extract for 63 d. Compared with the controls, (1) dietary supplementation with bilberry extract did not affect the growth performance of chickens from 1 to 63 d. (2) At 21 d, the relative weight of the bursa of Fabricius was increased (p < 0.05) by dietary supplementation with 400 mg/kg bilberry extract. Bilberry extract decreased the concentrations of IgY and IgM in blood plasma of 63-d chickens (p < 0.05). (3) For 21-d chickens, dietary supplementation with 400 mg/kg bilberry extract increased (p < 0.05) the activity of GSH-Px in blood plasma and jejunal mucosa (p < 0.05). Supplementation with 100 mg/kg bilberry extract increased (p < 0.05) the activities of T-SOD in jejunal mucosa and GSH-Px in the liver and decreased (p < 0.05) the MDA concentration in the liver. For chickens at the age of 63 d, both levels of bilberry extract increased activity of T-SOD in blood plasma (p < 0.05) and reduced MDA concentration in the jejunum (p < 0.05). (4) Supplementation with bilberry extract in the diet decreased the MDA concentration (B100) in muscle of 63-d chickens at 45 min postmortem and increased (p < 0.05) the activity of T-SOD (B400) at 4 d postmortem. (5) In breast muscle at 63 d, birds supplemented with bilberry extract (B400) had increased pH and drip loss while drip loss was reduced in the B100 treatment (p < 0.05); treatments did not affect inosinic acid or intramuscular fat contents. In conclusion, dietary supplementation of yellow-feathered chickens with bilberry extract enhanced the relative weight of the bursa of Fabricius, and broadly increased activities of antioxidant enzymes; indices of meat quality were improved without impact on growth performance. Considering the results in the current research, 100 mg/kg bilberry extract was recommended when supplemented in chickens.

High dietary copper induces oxidative stress and leads to decreased egg quality and reproductive performance of Chinese Yellow broiler breeder hens.

The objective of this study was to investigate the effects of dietary copper (Cu) on production, egg quality, and hatchability of Chinese Yellow broiler breeder hens and growth performance of their offspring. A total of 576 30-week-old hens were randomly allotted into 6 groups, each with 6 replicates (8 cages for each replicate with 2 birds per cage). The basal diet contained 3.50 mg/kg Cu, and the other 5 treatment diets contained 8.5, 13.5, 23.5 43.5, and 83.5 mg/kg Cu, respectively, additionally supplemented with Cu on the basal diet. The trial lasted for 15 wk. Qualified egg rate of birds fed 23.5 or 83.5 mg/kg Cu was significantly decreased (P < 0.05) compared with those given 3.5, 8.5, or 13.5 mg/kg Cu. Plasma malondialdehyde concentration showed quadratic effect (P = 0.002) which that decreased first then increased with dietary Cu increased. Highest values of Cu content and hepatic activity of Cu-ATPase occurred in hens fed 83.5 mg/kg dietary Cu with linear (P = 0.001) and quadratic (P = 0.001) effects. Shell strength and proportion on 18th day of live embryos of hens fed 13.5 mg/kg Cu were the greatest compared with other groups respectively (P < 0.05); rate of qualified eggs for hatch and hatchability of fertilized eggs of hens fed 83.5 mg/kg Cu were the least (P < 0.05). In conclusion, both inadequate (3.5 mg/kg diet) and excess (83.5 mg/kg) of dietary Cu can induce oxidative stress in hens and lead to decreased egg quality. Hatchability and growth performance of offspring were decreased when breeder hens were fed excess Cu in spite of greater hatching weight. The appropriate dietary Cu level for Chinese Yellow broiler breeder hens during the egg-laying period is 15.7 to 21.2 mg/kg (1.81-2.44 mg Cu fed per day) when based on Cu level and Cu-ATPase activity in the liver. This dietary Cu requirement is approximately doubled (∼40 mg/kg, ∼4.60 mg Cu per bird per day) for maximal response of eggshell thickness.

Virome and Blood Meal-Associated Host Responses in Ixodes persulcatus Naturally Fed on Patients.

The long-lasting co-evolution of ticks with pathogens results in mutual adaptation. Blood-feeding is one of the critical physiological behaviors that have been associated with the tick microbiome; however, most knowledge was gained through the study of laboratory-reared ticks. Here we detached Ixodes persulcatus ticks at different stages of blood-feeding from human patients and performed high-throughput transcriptomic analysis on them to identify their virome and genes differentially expressed between flat and fully fed ticks. We also traced bloodmeal sources of those ticks and identified bats and three other potential mammalian hosts, highlighting the public health significance. We found Jingmen tick virus and 13 putative new viruses belonging to 11 viral families, three of which even exhibited high genetic divergence from viruses previously reported in the same tick species from the same geographic region. Furthermore, differential expression analysis suggested a downregulation of antioxidant genes in the fully fed I. persulcatus ticks, which might be related to bloodmeal-related redox homeostasis. Our work highlights the significance of active surveillance of tick viromes and suggests a role of reactive oxygen species (ROS) in modulating changes in the microbiome during blood-feeding.

Potential Effects of Acidifier and Amylase as Substitutes for Antibiotic on the Growth Performance, Nutrient Digestion and Gut Microbiota in Yellow-Feathered Broilers.

This study was conducted to evaluate the effects of acidifier (benzoic acid, BA), amylase (AL) and their combination as substitutes for antibiotics on growth performance, antioxidation, nutrient digestion and gut microbiota of yellow-feathered broilers. A total of 1440 twenty-one-day-old broilers were randomly allocated to six treatments. Broilers in the control group (CON) were fed a basal diet, whereas birds in the other five groups were fed the basal diet supplemented with antibiotic (zinc bacitracin, AT, 40 mg/kg), BA (2000 mg/kg), low level AL (AL-L, 300 mg/kg), high level AL (AL-H, 500 mg/kg) and the combination of AL-H and BA (BA+AL-H). The experimental animals were killed at the end of the trial (21 day-63 day) then blood samples were collected from two birds per pen. Bird weight, feed intake and survival rate were recorded on pen basis. Growth performance was not significantly influenced by AT, BA, AL-L, AL-H or BA+AL-H. Plasma uric acid (UA) was decreased from CON by all treatments; the activity of AKP in plasma was also lowered by AT, BA, AL-H and BA+AL-H. Plasma activity of LDH was reduced by BA. In the jejunal mucosa, Na+K+-ATP activity was increased by BA, AL-L, AL-H and BA+AL-H. Mucosal activities of T-AOC and CAT were increased with AL-L and AT supplementation, respectively. Additionally, the relative abundance of Escherichia coli (E. coli) in the cecal contents was reduced by BA+AL-H and, with the exception of AL-H, all treatments increased the relative abundance of Lactobacillus. In conclusion, dietary AT, BA, AL-L, AL-H or BA+AL were effective in improving the antioxidant capacity, nutrient digestion and gut microbiota composition. No significant differences were observed in the tested variables between AT and other treatments, indicating that BA, AL and their combination may be alternatives to dietary inclusion of zinc bacitracin. Dietary addition of 500 mg/kg AL and 2000 mg/kg BA was an optimum supplementation dose.

Proteomic alteration of porcine intestinal epithelial cells after pretreatment with Lactobacillus plantarum followed by infection with enterotoxigenic Escherichia coli F4.

Enterotoxigenic Escherichia coli (ETEC) F4 causes diarrhea in infants and weaned piglets. The technique of isobaric tags for relative and absolute quantitation (iTRAQ) was used in this study to determine the differentially expressed proteins in porcine intestinal epithelial cells (IPEC-J2) after pretreatment with Lactobacillus plantarum (LP) followed by challenge with ETEC F4. A total of 4771 proteins were identified in IPEC-J2 cells, with 90, 105, and 134 differentially expressed proteins in cells exposed to ETEC, LP, and LP + ETEC, respectively. The COG analysis divided the identified proteins into 20 categories. The GO and KEGG annotation indicated that most of the differentially expressed proteins were enriched in various biological metabolism including cell cycle control, cell division and differentiation. Additionally, western blotting analyses confirmed the reduced abundance of selected proteins of the mTOR and MAPK signal pathways affected by ETEC F4. Moreover, LP pretreatment increased JNK activation in IPEC-J2 cells infected with ETEC F4. These results may provide further insights into the mechanisms involved in the interaction between ETEC F4 and intestinal epithelial cells, and broaden the understanding of the protective effects of LP in alleviating ETEC-provoked diarrhea of piglets.

Influence of Mushroom Polysaccharide, Nano-Copper, Copper Loaded Chitosan, and Lysozyme on Intestinal Barrier and Immunity of LPS-mediated Yellow-Feathered Chickens.

This study investigated the influence of dietary supplementation with some antibiotic alternatives on growth performance, intestinal barrier, and immunity of lipopolysaccharide (LPS) challenged chicks. Wenshi females, aged 4 days, were allocated randomly into eight groups, each with six replicates of 20 birds (n = 120/treatment), which received a basal diet supplemented with 0 (control), 0 (LPS), 200 mg/kg aureomycin, 50 mg/kg mushroom polysaccharide, 100 mg/kg mushroom polysaccharide, 500 mg/kg nano-copper, 300 mg/kg copper loaded chitosan, and 500 mg/kg lysozyme for 21 days. On day 18 and 20, the control birds were injected with 0.5 mL saline solution, the other treatments were injected with 0.5 mL saline containing 500 µg LPS/kg body weight (BW). The results indicated that LPS treatment reduced the BW, average daily gain (ADG), and daily feed intake (ADFI) than the controls (p < 0.05), and the antibiotic and the tested alternatives could not retrieve the normal BW, ADG, and ADFI. The tested additives reduced several negative effects of LPS; they reduced diamine oxidase activity and inflammatory mediators in plasma, jejunal mucosa, spleen and thymus, increased content of immunoglobulin in plasma and jejunal mucosa, and decreased gene expression of inducible nitric oxide synthase and Cyclooxygenase 2 in jejunal mucosa.

Effects of maternal and dietary vitamin A on growth performance, meat quality, antioxidant status, and immune function of offspring broilers.

The aim of this study was to evaluate the effects of maternal and dietary vitamin A (VA) level on growth performance, meat quality, antioxidant status, and immune function of offspring broilers. Chinese yellow-feathered breeder hens were fed a basal diet supplemented with 0, 5,400, 10,800, and 21,600 IU/kg VA for 8 wk, with 6 replicates of 22 hens per replicate. Then the offspring hatched from each of the 4 maternal groups were fed a basal diet supplemented with 0 or 5,000 IU/kg VA for 63 D. Overall, there were 8 treatment combinations, each with 6 replicate pens of 20 birds. Results showed that (1) providing VA in offspring diets increased final body weight (FW), average daily gain, and average daily feed intake but reduced feed-to-gain ratio and mortality of offspring broilers (P < 0.05), whereas maternal provision of VA did not significantly affect the growth performance and mortality of offspring broilers. Maternal or offspring VA did not affect proportion of breast or thigh muscle (P > 0.05). (2) Maternal feeding with 21,600 IU/kg VA increased (P < 0.05) pH 24 h postmortem of breast muscle, compared with those without maternal supplication of VA. Dietary provision of 5,000 IU/kg VA in the posthatching diet decreased (P < 0.05) drip loss, yellowness (b∗) value and lightness (L∗) value, and increased shear force and pH of breast muscle compared with those without dietary VA supplication. (3) Maternal or offspring VA did not affect the activities of total superoxide dismutase and glutathione peroxidase (GSH-Px) or the content of malondialdehyde; however, there was a significant interaction (P < 0.05) between maternal and offspring VA on the activity of GSH-Px in serum. (4) Dietary provision of 5,000 IU/kg VA increased (P < 0.05) the weight proportion of liver and bursa of fabricius, whereas maternal feeding with 21,600 IU/kg VA increased the hatchling BW. Maternal feeding with 5,400 and 21,600 IU/kg VA decreased (P < 0.05) splenic interferon-γ (IFN-γ) transcripts and increased (P < 0.05) those of interleukin-2 (IL-2) in the progeny. There were interactions (P < 0.05) between maternal and offspring VA on splenic IL-2, IL-1ß, and IFN-γ expression. In summary, maternal and offspring provision of VA both had influence on meat quality and immune function in progeny broilers. Dietary VA increased growth performance, whereas the maternal VA affected the initial body weight of progeny when hatched, but the difference in performance caused by maternal VA level was able to be eliminated by dietary VA supplementation. Therefore, offspring provision had greater importance than maternal VA in the production; however, both should be considered in broiler nutrition to achieve good meat quality and immune status of broilers.

Protection of Porcine Intestinal-Epithelial Cells from Deoxynivalenol-Induced Damage by Resveratrol via the Nrf2 Signaling Pathway.

Deoxynivalenol (DON), a common mycotoxin, usually induces oxidative stress and affects the intestinal health of humans and animals. This study investigated the protective effect of resveratrol (RES), a natural antioxidant, on alleviating the cytotoxicity induced by DON in the porcine intestinal-epithelial cell line (IPEC-J2). Cells were incubated with RES for 24 h and then exposed to DON for another 24 h. Cell viability, proliferation, apoptosis, and oxidative-stress indicators were determined. In comparison with DON-only-treated cells, pretreatment with RES (15 µM) increased the cell viability (79.74 ± 2.02 vs 90.98 ± 2.66%, P < 0.01), improved proliferation (EdU-positive cells, 26.42 ± 1.12 vs 32.05 ± 0.78%, P < 0.01), decreased accumulation of intracellular reactive oxygen species (ROS, 1.68 ± 0.05 vs 1.29 ± 0.06, P < 0.01), stabilized mitochondrial-membrane potential (MMP, 8.98 ± 1.40 vs 2.29 ± 0.76, P < 0.001), and prevented apoptosis induced by DON (13.91 ± 1.20 vs 6.83 ± 0.52%, P < 0.01). RES activated the Nrf2 signaling pathway, and transfection with Nrf2 siRNA abrogated the protection of RES against DON-induced cytotoxicity, accumulation of intracellular ROS, and mitochondria-dependent apoptosis. Collectively, RES protects IPEC-J2 cells against DON-induced damage at least partly via the Nrf2 signaling pathway.

mTORC1 Mediates Lysine-Induced Satellite Cell Activation to Promote Skeletal Muscle Growth.

As the first limiting amino acid, lysine (Lys) has been thought to promote muscle fiber hypertrophy by increasing protein synthesis. However, the functions of Lys seem far more complex than that. Despite the fact that satellite cells (SCs) play an important role in skeletal muscle growth, the communication between Lys and SCs remains unclear. In this study, we investigated whether SCs participate directly in Lys-induced skeletal muscle growth and whether the mammalian target of rapamycin complex 1 (mTORC1) pathway was activated both in vivo and in vitro to mediate SC functions in response to Lys supplementation. Subsequently, the skeletal muscle growth of piglets was controlled by dietary Lys supplementation. Isobaric tag for relative and absolute quantitation (iTRAQ) analysis showed activated SCs were required for longissimus dorsi muscle growth, and this effect was accompanied by mTORC1 pathway upregulation. Furthermore, SC proliferation was governed by medium Lys concentrations, and the mTORC1 pathway was significantly enhanced in vitro. After verifying that rapamycin inhibits the mTORC1 pathway and suppresses SC proliferation, we conclude that Lys is not only a molecular building block for protein synthesis but also a signal that activates SCs to manipulate muscle growth via the mTORC1 pathway.