RESUMO
Currently, there are plenty of histochemical methods to classify pig muscle fibers, which confused the naming and classification of muscle fibers. This study aims to analyze the difference and correlation of 6 different histochemical methods and select the most suitable method for muscle fiber classification at the molecular and histomological levels by in-situ RT-PCR and enzyme histochemical methods. Muscle fiber samples, including psoas (PM), semitendinosus (SM) and trapezius muscle (TM), were collected from Large Spotted (LS), Lantang (LT) and Landrace (LR) pigs at their market-ages (LS at 150 d, LT at 210 d, and LR at 150 d). 6 kinds of histochemical methods combining actomyosin adenosine triphosphatase (AM-ATPase) with succinate dehydrogenase (SDH) enzyme were conducted to differentiate fiber types. 2 types of fibers (I and II) were differentiated by acid 2-fibre (2-AC) or alkaline 2-fibre classification(2-AL), 3 types of fibers (ßR, αR and αW) by 3-AC or 3-AL, and 4 types of fibers (I, IIa, IIx and IIb) by 4-AC, or 4-AL. Results showed that AC and AL muscle-fiber classification were consistent in reflecting the characteristics of muscle fibers(P > 0.05), but the color of each muscle fiber type was just opposite. AC methods may be superior to AL methods because of their clear staining background, the sensitivity to staining condition. But there were breed differences and tissue specificity in the optimal preincubation condition. The optimal acid preincubation condition for classifying muscle fibers was pH4.30 for LT, while pH 4.35 for the LS and LR pigs. Meanwhile the optimal acid preincubation condition was pH4.35 for PM, while pH4.40 for TM or SM. For further selection from 2, 3, 4-AC, in-situ RT-PCR was applied to detect the mRNA distribution of myosin heavy chain I (MyHC-I). By combining in-situ PCR with enzyme histochemistry methods, MyHC-I gene and its product - Type I fibrocytes were directly located in cells at both molecular level and morphological level. Compared with the cross-sectional area (CSA) of different muscle fibers (i.e. I, II, ßR, αR, αW, IIa, IIx and IIb) identified by enzyme histochemistry, it was found that the CSAs with stronger mRNA expression signal of MyHC-â were closer to those of the Type I muscle fiber measured by 4-AC enzyme histochemistry (P > 0.05). Therefore, 4-AC may be considered as the most proper muscle typing method to study muscle fiber typing as well as meat quality. And the combination of in-situ RT-PCR and histochemistry may help better understand porcine muscle fiber characteristics and meat quality in pigs.
RESUMO
Introduction: Feeding bamboo powder is a kind of fiber raw material mainly composed of insoluble dietary fiber (IDF). In this study, IDF-based rice husk meal and feeding bamboo powder were used to compare the effects of bamboo fiber on fecal microflora and the performance of lactating sows and their offspring piglets. Methods: Thirty healthy crossbred gilts (Yorkshire × Landrace) at day 105 of gestation were randomly allocated into three groups: CON, TRE1 supplemented with 2% BBF1 (feeding bamboo powder), and TRE2 supplemented with 2% BBF2 (99% feeding bamboo powder +1% bamboo fiber polymer material). The reproductive performance, serum indexes, and fecal microbiota of sows and piglets were analyzed. The results showed that, compared with CON, the average feed intake of sows in TRE1 during the second week of lactation was significantly increased by 21.96% (p < 0.05), the average daily gain (ADG) per litter in TRE1 on 11-21 days and 3-21 days of lactation was significantly increased by 50.68 and 31.61%, respectively (p < 0.05), and the serum triglyceride content of sows in TRE1 on the 21st day of lactation was significantly increased (p < 0.05). The 16S rRNA analysis showed that dietary bamboo fiber significantly increased the fecal microbial richness index Ace, Chao, and Sobs of sows (p < 0.05) and tended to increase the Sobs index of suckling piglets on day 21 (p < 0.10). Compared with CON, BBF1 supplementation significantly decreased the abundance of Christensenellaceae_R-7_group in feces of sows on days 7 and 21 after delivery (p < 0.05), while BBF2 decreased the genera Christensenellaceae_R-7_group on days 7 (p < 0.10) and 21 (p < 0.05) after delivery. Spearman correlation analysis showed that the abundance of Phascolarctobacterium in the feces of piglets on the 21st day after delivery was significantly positively correlated with diarrhea rate and significantly negatively correlated with ADG per litter, day 21 litter weight, and 3- to 21-day survival rate. In contrast, Christensenellaceae_R-7_group was significantly negatively correlated with diarrhea rate and positively correlated with ADG per litter. Discussion: These results indicated that maternal BBF1 supplementation improved the litter weight gain of suckling piglets, which was associated with the improvement of diversity and structure of the fecal microbiota in the piglets.
RESUMO
Xylooligosaccharide (XOS) is known as a prebiotic, however, it is unknown whether XOS can directly protect against bacterial infection. This study aimed to investigate the direct inhibitory effects of XOS on Salmonella Typhimurium colonization and the inductive impairments in gut health and growth performance in broilers. We first probed the inhibitory effects of XOS on S. Typhimurium adhesion and its induction of intestinal epithelial cell (IPEC-J2) injuries. Afterward, 168 one-day-old yellow-feathered broilers were randomly divided into 3 groups (7 replicates/group): negative control (NC, received a basal diet), positive control (PC, received a basal diet with S. Typhimurium challenge) and XOS group (PC birds + 1,500 mg/kg XOS). All birds except those in NC were orally challenged with S. Typhimurium from 8 to 10 d of age. Parameters were analyzed on d 11. The results showed that XOS inhibited S. Typhimurium adhesion and the inductive injuries of IPEC-J2 cells by lowering (P < 0.05) certain adhesion-related genes expression of this bacterium. It also alleviated S. Typhimurium-induced increase (P < 0.05) in the expression of certain inflammatory cytokines and tight junction (TJ) proteins of IPEC-J2 cells. Supplementing XOS to S. Typhimurium-challenged broilers attenuated the elevations (P < 0.05) in S. Typhimurium colonization of ileal mucosa and its translocation to the liver and spleen, as well as increased (P < 0.05) certain TJ proteins expression of ileum. Besides, XOS addition normalized S. Typhimurium-induced impairments (P < 0.05) in ileal morphology, final body weight and average daily gain in broilers. Collectively, supplemental XOS directly suppressed intestinal colonization of S. Typhimurium by diminishing its adhesiveness and subsequently mitigated destructions in intestinal barriers, thus contributing to weaken growth retardation in challenged broilers. Our findings provide a new insight into the mechanisms of XOS limiting Salmonella infection in chickens.
Assuntos
Salmonelose Animal , Salmonella typhimurium , Animais , Galinhas , Salmonelose Animal/prevenção & controle , Salmonelose Animal/microbiologia , Dieta/veterináriaRESUMO
This study aimed to investigate the potential mitigating effects of N-acyl homoserine lactonase (AHLase) on the virulence of Salmonella typhimurium and its induction of intestinal damages in broilers. In vitro study was firstly conducted to examine if AHLase treatment could attenuate the virulence of S. typhimurium. Then, an in vivo experiment was performed by allocating 240 broiler chicks at 1 d old into 3 groups (8 replicates per group): negative control (NC), positive control (PC), and PC supplemented with 10,000 U/kg AHLase. All chicks except those in NC were orally challenged by S. typhimurium from 8 to 10 d of age. Parameters were measured on d 11 and 21. The results showed that treatment with 1 U/mL AHLase suppressed the biofilm-forming ability (including biofilm biomass, extracellular DNA secretion and biofilm formation-related gene expression), together with swarming motility and adhesive capacity of S. typhimurium. Supplemental 10,000 U/kg AHLase counteracted S. typhimurium-induced impairments (P < 0.05) in broiler growth performance (including final body weight, average daily gain and average daily feed intake) during either 1-11 d or 12-21 d, and increases (P < 0.05) in the indexes of liver, spleen and bursa of Fabricius on d 11, together with reductions (P < 0.05) in ileal villus height and its ratio to crypt depth on both d 11 and 21. AHLase addition also normalized the increased (P < 0.05) mRNA expression of ileal occludin on both d 11 and 21 in S. typhimurium-challenged broilers. However, neither S. typhimurium challenge nor AHLase addition altered (P > 0.05) serum diamine oxidase activity of broilers. Noticeably, S. typhimurium challenge caused little change in the mRNA expression of ileal inflammatory cytokines except for an increase (P < 0.05) in interleukin-8 expression on d 11, whereas AHLase addition normalized (P < 0.05) this change. In conclusion, AHLase treatment could attenuate the virulence and pathogenicity of S. typhimurium, thus contributing to alleviate S. typhimurium-induced growth retardation and intestinal damages in broilers.
RESUMO
Salmonella enterica serovar typhimurium (S. Typhimurium) is a common Gram-negative foodborne pathogenic bacterium that causes gastrointestinal disease in humans and animals. It is well known that adhesins and invasins play crucial roles in the infection mechanism of S. Typhimurium. S. Typhimurium STM0306 has been denoted as a putative protein and its functions have rarely been reported. In this study, we constructed the STM0306 gene mutant strain of S. Typhimurium and purified the recombinant STM0306 from Escherichia coli. Deletion of the STM0306 gene resulted in reduced adhesion and invasion of S. Typhimurium to IPEC-J2, Caco-2, and RAW264.7 cells. In addition, STM0306 could bind to intestinal epithelial cells and induced F-actin modulation in IPEC-J2 cells. Furthermore, we found that STM0306 activated the nuclear factor kappa B (NF-κB) signaling pathway and increased the mRNA expression of pro-inflammatory cytokines such as IL-1ß, TNF-α, as well as chemokine CXCL2, thus resulting in cellular inflammation in host cells. In vivo, the deletion of the STM0306 gene led to reduced pathogenicity of S. Typhimurium, as evidenced by lower fecal bacterial counts and reduced body weight loss in S. Typhimurium infected mice. In conclusion, the STM0306 of S. Typhimurium is an important adhesin/invasin involved in the pathogenic process and cellular inflammation of the host.
Assuntos
Adesinas Bacterianas , Salmonella typhimurium , Humanos , Animais , Camundongos , Salmonella typhimurium/metabolismo , Sorogrupo , Células CACO-2 , Adesinas Bacterianas/genética , Adesinas Bacterianas/metabolismo , Inflamação/genéticaRESUMO
Introduction: This study was conducted to explore the effects of supplementary feeding of bamboo powder on the physical parameters of sows during the perinatal period of 7 days ± in parturition, including farrow duration, serum biochemical indexes, fecal physicochemical indexes, and microbial flora. Methods: Thirty pregnant sows were randomly divided into three groups: the control group was fed a basal diet, TRE1 group and TRE2 group were fed a basal diet supplemented with 30 g d-1 and 60 g d-1 bamboo powder, respectively. Multiple parameters of sows and offspring piglets were determined. Results: The contents of serum total cholesterol and triglyceride of sows in TRE2 group were significantly lower than those in the control group. The contents of serum malondialdehyde of sows in TRE2 and TRE1 groups were significantly lower than that in control group. The water content of sow feces in TRE2 group was significantly higher than that in control group, and the pH values of sows in TRE2 and TRE1 groups were significantly higher than that in control group. The richness index (Chao) of sow fecal bacterial community in TRE2 group was significantly lower than that of the control group, and the Ace and Sobs indexes tended to be lower than those of the control group. At the phylum level, the relative abundance of Actinobacteriota in the feces of sows in TRE2 group was significantly lower than that of the control group, while that of Fusobacteriota in the feces of suckling piglets in TRE2 group tended to be lower than that of the control group. At the genus level, among the Top10 dominant bacteria, the relative abundance of Tissierella in the feces of sows in TRE2 group was significantly lower than that of the control group while that of Fusobacterium in the feces of suckling piglets in TRE2 group tended to be lower than that of the control group. The relative abundance of Clostridium_sensu_stricto_1, Terrisporobacter, Turicibacter, and Tissierella in the feces of sows in TRE2 group was significantly lower than that of TRE1 group (p < 0.05), while Lactobacillus tended to be higher than that of TRE1 group (p < 0.10). Discussion: The results suggested that supplementary feeding 60 g d-1 bamboo powder could increase the water content in the feces of sows, reduce the oxidative damage, and tend to reduce the relative abundance of opportunistic pathogenic Fusobacterium for suckling piglets, while it reduced the fecal microbial diversity of sows.
RESUMO
MicroRNAs are one of the key determinants of muscle fibre development and phenotype in mammals. The preliminary experiment implied that microRNA-27a (miR-27a) might involve in regulation of muscle fibre type composition of pigs. Thereby, the present study aimed to confirm the regulatory effect of miR-27a on porcine type I muscle fibre-encoding gene (myosin heavy chain gene 7, MYH7) expression and its related mechanism. We firstly observed opposite expression patterns between miR-27a and MYH7 as well as between miR-27a and peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α) during differentiation of porcine skeletal muscle satellite cells. Through the subsequent transfection analysis in porcine myotubes, we found that miR-27a suppressed the expression of MYH7 and PGC-1α. Besides, miR-27a induced inhibition of PGC-1α downstream targets, namely myocyte enhancer factor-2C (MEF2C) along with mitochondrial biogenesis and oxidative metabolism-related factors such as nuclear respiratory factor 1 (NRF-1), mitochondrial transcription factor A (mtTFA), cytochrome c (Cytc) and cytochrome oxidase IV (COX â £) and succinodehydrogenase (SDH). Dual-luciferase reporter analysis revealed that miR-27a could bind to the predicted target site in the 3'-untranslated regions of PGC-1α mRNA, confirming a direct targeting of PGC-1α by miR-27a. Moreover, PGC-1α silencing abolished the promotive effects of miR-27a inhibitor on MYH7, PGC-1α and its downstream targets (MEF2C, NRF-1, mtTFA, COX â £, Cytc and SDH) in porcine myotubes. Collectively, miR-27a inhibits porcine MYH7 expression by negatively regulating PGC-1α and PGC-1α-controlled MEF2C expression as well as mitochondrial biogenesis and oxidative metabolism. Our findings may provide a molecular target for genetic or nutritional control of muscle fibre phenotype of pigs, probably having an important implication for regulating pork quality.
Assuntos
MicroRNAs , PPAR gama , Suínos , Animais , PPAR gama/genética , MicroRNAs/genética , MicroRNAs/metabolismo , Fibras Musculares Esqueléticas/metabolismo , Expressão Gênica , Músculo Esquelético/metabolismo , Mamíferos/metabolismoRESUMO
BACKGROUND: Salmonella Typhimurium challenge causes a huge detriment to chicken production. N-acyl homoserine lactonase (AHLase), a quorum quenching enzyme, potentially inhibits the growth and virulence of Gram-negative bacteria. However, it is unknown whether AHLase can protect chickens against S. Typhimurium challenge. This study aimed to evaluate the effects of AHLase on growth performance and intestinal health in broilers challenged by S. Typhimurium. A total of 240 one-day-old female crossbred broilers (817C) were randomly divided into 5 groups (6 replicates/group): negative control (NC), positive control (PC), and PC group supplemented with 5, 10 or 20 U/g AHLase. All birds except those in NC were challenged with S. Typhimurium from 7 to 9 days of age. All parameters related to growth and intestinal health were determined on d 10 and 14. RESULTS: The reductions (P < 0.05) in body weight (BW) and average daily gain (ADG) in challenged birds were alleviated by AHLase addition especially at 10 U/g. Thus, samples from NC, PC and PC plus 10 U/g AHLase group were selected for further analysis. S. Typhimurium challenge impaired (P < 0.05) intestinal morphology, elevated (P < 0.05) ileal inflammatory cytokines (IL-1ß and IL-8) expression, and increased (P < 0.05) serum diamine oxidase (DAO) activity on d 10. However, AHLase addition normalized these changes. Gut microbiota analysis on d 10 showed that AHLase reversed the reductions (P < 0.05) in several beneficial bacteria (e.g. Bacilli, Bacillales and Lactobacillales), along with increases (P < 0.05) in certain harmful bacteria (e.g. Proteobacteria, Gammaproteobacteria, Enterobacteriaceae and Escherichia/Shigella) in PC group. Furthermore, AHLase-induced increased beneficial bacteria and decreased harmful bacteria were basically negatively correlated (P < 0.05) with the reductions of ileal IL-1ß and IL-8 expression and serum DAO activity, but positively correlated (P < 0.05) with the increased BW and ADG. Functional prediction revealed that AHLase abolished S. Typhimurium-induced upregulations (P < 0.05) of certain pathogenicity-related pathways such as lipopolysaccharide biosynthesis, shigellosis, bacterial invasion of epithelial cells and pathogenic Escherichia coli infection of gut microbiota. CONCLUSIONS: Supplemental AHLase attenuated S. Typhimurium-induced growth retardation and intestinal disruption in broilers, which could be associated with the observed recovery of gut microbiota dysbiosis.
RESUMO
Catalase (CAT) can eliminate oxygen radicals, but it is unclear whether exogenous CAT can protect chickens against deoxynivalenol (DON)-induced oxidative stress. This study aimed to investigate the effects of supplemental CAT on antioxidant property and gut microbiota in DON-exposed broilers. A total of 144 one-day-old Lingnan yellow-feathered male broilers were randomly divided into three groups (six replicates/group): control, DON group, and DON + CAT (DONC) group. The control and DON group received a diet without and with DON contamination, respectively, while the DONC group received a DON-contaminated diet with 200 U/kg CAT added. Parameter analysis was performed on d 21. The results showed that DON-induced liver enlargement (p < 0.05) was blocked by CAT addition, which also normalized the increases (p < 0.05) in hepatic oxidative metabolites contents and caspase-9 expression. Additionally, CAT addition increased (p < 0.05) the jejunal CAT and GSH-Px activities coupled with T-AOC in DON-exposed broilers, as well as the normalized DON-induced reductions (p < 0.05) of jejunal villus height (VH) and its ratio for crypt depth. There was a difference (p < 0.05) in gut microbiota among groups. The DON group was enriched (p < 0.05) with some harmful bacteria (e.g., Proteobacteria, Gammaproteobacteria, Enterobacteriales, Enterobacteriaceae, and Escherichia/Shigella) that elicited negative correlations (p < 0.05) with jejunal CAT activity, and VH. DONC group was differentially enriched (p < 0.05) with certain beneficial bacteria (e.g., Acidobacteriota, Anaerofustis, and Anaerotruncus) that could benefit intestinal antioxidation and morphology. In conclusion, supplemental CAT alleviates DON-induced oxidative stress and intestinal damage in broilers, which can be associated with its ability to improve gut microbiota, aside from its direct oxygen radical-scavenging activity.
Assuntos
Galinhas , Microbioma Gastrointestinal , Animais , Masculino , Galinhas/metabolismo , Catalase/metabolismo , Disbiose/veterinária , Antioxidantes/farmacologia , Antioxidantes/metabolismo , Estresse Oxidativo , Dieta/veterinária , Suplementos Nutricionais/análise , Ração Animal/análiseRESUMO
Glucose oxidase (GOD) could benefit intestinal health and growth performance in animals. However, it is unknown whether GOD can protect piglets against bacterial challenge. This study aimed to evaluate the protective effects of GOD on growth performance, clinical symptoms, serum parameters, and intestinal health in piglets challenged by enterotoxigenic Escherichia coli (ETEC). A total of 44 male weaned piglets around 38 days old were divided into four groups (11 replicates/group): negative control (NC), positive control (PC), CS group (PC piglets +40 g/t colistin sulfate), and GOD group (PC piglets +200 g/t GOD). All piglets except those in NC were challenged with ETEC (E. coli K88) on the 11th day of the experiment. Parameter analysis was performed on the 21st day of the experiment. The results showed that the ETEC challenge elevated (p < 0.05) the rectal temperature and fecal score of piglets at certain time-points post-challenge, reduced (p < 0.05) serum glucose and IgG levels but increased (p < 0.05) serum alanine aminotransferase activity, as well as caused (p < 0.05) intestinal morphology impairment and inflammation. Supplemental GOD could replace CS to reverse (p < 0.05) the above changes and tended to increase (p = 0.099) average daily gain during the ETEC challenge. Besides, GOD addition reversed ETEC-induced losses (p < 0.05) in several beneficial bacteria (e.g., Lactobacillus salivarius) along with increases (p < 0.05) in certain harmful bacteria (e.g., Enterobacteriaceae and Escherichia/Shigella). Functional prediction of gut microbiota revealed that ETEC-induced upregulations (p < 0.05) of certain pathogenicity-related pathways (e.g., bacterial invasion of epithelial cells and shigellosis) were blocked by GOD addition, which also normalized the observed downregulations (p < 0.05) of bacterial pathways related to the metabolism of sugars, functional amino acids, nucleobases, and bile acids in challenged piglets. Collectively, GOD could be used as a potential antibiotic alternative to improve growth and serum parameters, as well as attenuate clinical symptoms and intestinal disruption in ETEC-challenged piglets, which could be associated with its ability to mitigate gut microbiota dysbiosis. Our findings provided evidence for the usage of GOD as an approach to restrict ETEC infection in pigs.
RESUMO
OBJECTIVE: This study was aimed to explore the efficacy of combination of endo-xylanase (Xyn) and xylan-debranching enzymes (arabinofuranosidase, Afd and feruloyl esterase, FE) in improving utilization of bran in piglet diet. METHODS: In vitro experiments were firstly conducted to examine the enzymological properties of Xyn, Afd, and FE, concurrent with their effect on degradation of arabinoxylan (Abx) in bran. In vivo experiment was then implemented by allocating two hundred and seventy 35-d-old postweaning piglets into 3 groups (6 replicates/group), which received bran-containing diet supplemented with Xyn (1,600 U/kg) or its combination with Afd (0.8 U/kg) and FE (4 U/kg) or without enzyme. RESULTS: Both Xyn, Afd, and FE are relatively stable against the changes in temperature and pH value. Combining Xyn with Afd and FE had a superiority (p<0.05) over Xyn alone and its combination with Afd or FE in promoting (p<0.05) degradation of Abx in different brans. Combined treatment with Xyn, Afd, and FE was more beneficial than Xyn alone to induce increasing trends (p<0.10) of average daily gain, final body weight and feed efficiency of piglets fed bran-containing diet. Moreover, combination of Xyn, Afd, and FE showed advantages (p<0.05) over Xyn alone in causing reductions (p<0.05) in diarrhea rate and cecal pH value, concurrent with increases (p<0.05) in cecal and colonic acetic acid and total volatile fatty acid concentrations, as well as cecal butyric acid concentration of piglets fed bran-containing diet. CONCLUSION: Combining Xyn with Afd and FE was more beneficial than Xyn alone in promoting degradation of Abx in bran, along with growth performance and intestinal volatile fatty acid profile of piglets received bran-containing diet. Thereby, combination of Xyn, Afd, and FE had a superior efficacy relative to Xyn alone in improving application of cereal bran in piglet diet.
RESUMO
Adding insoluble fiber to diet of broilers has been reported to improve intestinal health and promote growth performance. Bamboo powder is a cheap raw material with rich insoluble fiber. This study aims to explore the effects of feeding micronized bamboo powder (MBP) on growth performance, serum biochemical indexes, intestinal microflora, and metabolism of broilers. A total of 1440 1-day-old slow-growing Ephedra chickens were randomly divided into three groups considering gender and body weight: (1) Group D: feeding with basal diet without antibiotics; (2) Group E: feeding with basal diet supplemented with 5% rice bran (RB); (3) Group F: feeding with basal diet supplemented with 1% MBP. Each group involved 8 replicates feeding for 22 days, with 60 chickens per replicate. Various indexes were detected. For the growth performance, the weight gain and feed consumption ratio (G: F) of Group F supplemented with MBP is 0.57 ± 0.04, which is significantly higher than that of E group supplemented with RB (0.52 ± 0.01, P < 0.05). For the serum biochemical indexes, the glutathione peroxidase activity in Group F is significantly higher than that of Group D, while the malondialdehyde content is significantly lower than that of Group D and Group E (P < 0.05 for all). The fresh cecal chyme is taken for determination. In Group F, the α diversity index Faith_pd is significantly lower in Group F than that of Group D. The microorganism species in cecal chyme of Group F and Group E are also different. The metabolic pathways of Group F, mainly in fatty acid metabolism, amino acid metabolism and intestinal immune IgA production, were different from those of Group D and Group E. Adding 1% MBP to broiler diet can enhance the anti-oxidant capacity, improve chyme microflora, regulate the metabolism pathways responsible for intestinal fatty acids, amino acids, and immunity.
Assuntos
Galinhas , Microbioma Gastrointestinal , Ração Animal/análise , Fenômenos Fisiológicos da Nutrição Animal , Animais , Antioxidantes/farmacologia , Ceco , Dieta/veterinária , Suplementos Nutricionais/análise , PósRESUMO
Salmonella Typhimurium is a common pathogen infecting the gastrointestinal tract of humans and animals, causing host gastroenteritis and typhoid fever. Heat shock protein (HtpG) as a molecular chaperone is involved in the various cellular processes of bacteria, especially under environmental stress. However, the potential association of HtpG with S. Typhimurium infection remains unknown. In this study, we clarified that HtpG could also play a role as an effector in S. Typhimurium infection. RNA-seq indicated that the flagellar assembly pathway, infection pathway, and chemotaxis pathway genes of S. Typhimurium were downregulated after the mutation of HtpG, which resulted in compromises of S. Typhimurium motility, biofilm formation, adhesion, invasion, and inflammation-inducing ability. In addition, HtpG recombinant protein was capable of promoting the proliferation of S. Typhimurium in host cells and the resultant inflammation. Collectively, our results illustrated an important role of HtpG in S. Typhimurium infection.
Assuntos
Proteínas de Choque Térmico , Salmonella typhimurium , Animais , Proteínas de Bactérias/genética , Proliferação de Células , Humanos , Salmonella typhimurium/genéticaRESUMO
This study investigated the effects of different amounts of wheat aleurone (WA) (0, 15%, 30%) inclusion in gestation diets on the reproductive performance, postprandial satiety, stress status and stereotypic behaviors of sows. A total of 84 Landrace × Yorkshire sows (parity 4.87 ± 1.32) at breeding were randomly allotted to one of the three isoenergetic and isonitrogenous dietary treatments based on parity and body weight. The results showed that, compared with the control (0), sows fed the WA diet had a higher serum concentration of peptide YY (PYY) (P < 0.05) and glucagon like peptide-1 (GLP-1) (P < 0.05) and a lower concentration of saliva cortisol (P < 0.01). Importantly, compared with the control group, only the 15% WA group had a higher concentration of the total antioxidant capacity (T-AOC) (P < 0.05), lower proportions of sitting (P = 0.05) and stillbirth rates (P < 0.01). Accordingly, the production cost per piglet born alive ($ 6.9 vs. $ 7.6) or per piglet born healthy ($ 7.4 vs. $ 7.9) declined in the 15% WA group versus the control group. Overall, 15% WA inclusion in gestation diets contributed to enhancing postprandial satiety, alleviating stress status and decreasing stillbirth rate of sows. This study provides a reference for the application of WA as a partial substitute for conventional feed ingredients to improve sows' reproductive performance.
RESUMO
Guar gum-derived galactomannan (GGGM) has been widely used in the food industry for a long time and its adverse impacts have been scarcely reported. Galactomannan is considered to have a structure similar to the surface components of certain pathogens, and the present study was thus conducted to investigate if oral administration of GGGM could cause physiological effects that were hypothesized to be related to intestinal inflammatory responses. The results showed that oral administration of GGGM resulted in compromises on growth performance, an increase of the relative weight of spleen and epididymal fat, and an elevation of the α1-acid glycoprotein content in both serum and livers of mice. With regard to energy metabolism-related indices, the activities of intestinal lactic dehydrogenase and succinic dehydrogenase were all increased by the GGGM treatment in both in vivo and in vitro experiments, the latter of which also showed an elevation in the consumption of reducing sugar by intestinal epithelial cells along with a reduced viability of these cells in response to the GGGM treatment. Notably, the GGGM treatment triggered intestinal inflammatory responses that were evidenced by the increased expression of intestinal inflammatory cytokines such as TNF-α and IL-6 both in vivo and in vitro, which were at least partially responsible for the increased energy expenditure in the intestine and the retardation of growth. The results of this study could expand our knowledge of GGGM administration and provide integrated insights into the consumption of GGGM-containing foods.
Assuntos
Metabolismo Energético/efeitos dos fármacos , Galactanos/farmacologia , Galactose/análogos & derivados , Inflamação/induzido quimicamente , Intestinos/efeitos dos fármacos , Intestinos/metabolismo , Mananas/farmacologia , Gomas Vegetais/farmacologia , Animais , Modelos Animais de Doenças , Galactanos/química , Galactose/química , Galactose/farmacologia , Masculino , Mananas/química , Camundongos , Gomas Vegetais/químicaRESUMO
Eucalyptus leaf polyphenols extract (EPE) has been proved to have various bioactivities, but few reports focus on its antioxidant mechanism in vivo. The purpose of this study was to elucidate the effect and mechanism of EPE dietary supplements on antioxidant capacity in chicken. A total of 216 chickens were randomly selected for a 40-day experiment. Four treatment groups received diets including the control diet only, the control diet + low EPE (0.6 g/kg), the control diet + moderate EPE (0.9 g/kg), and the control diet + high EPE (1.2 g/kg). Compared with control group, the glutathione peroxidase (GSH-Px) activity and glutathione (GSH) content in the breast muscle of the moderate EPE treatment group was significantly higher (p < 0.05), while the malonaldehyde (MDA) content in the moderate EPE group was reduced (p < 0.05). Moreover, proteomic and transcriptomic analyses of the breast muscle revealed that glutathione metabolism and the peroxisome were the two crucial metabolic pathways responsible for increased antioxidant capacity of the muscle. Accordingly, nine candidate genes and two candidate proteins were identified related to improved antioxidant status induced by EPE supplements. This research provides new insights into the molecular mechanism of antioxidant capacity in chickens treated with EPE dietary supplements.
Assuntos
Antioxidantes/metabolismo , Galinhas/genética , Eucalyptus/química , Perfilação da Expressão Gênica , Extratos Vegetais/farmacologia , Polifenóis/farmacologia , Proteoma/metabolismo , Animais , Galinhas/sangue , Suplementos Nutricionais , Regulação da Expressão Gênica/efeitos dos fármacos , Ontologia Genética , Anotação de Sequência Molecular , Músculos/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , RNA-SeqRESUMO
Inclusion of fiber in gestation diets is a method for enhancing satiety and reducing abnormal behaviors in restricted feeding sows without providing excess energy. The purpose of this study was to use an in vitro-in vivo method to appraise the effects of two available unconventional dietary fiber resources during gestation on sows' physio-chemical properties of diets, postprandial satiety, performance, abnormal behaviors, stress status and lactation feed intake under three different dietary treatments: control diet (CON diet), 5% resistant starch diet (RS diet), and 5% fermented soybean fiber diet (FSF diet) with a total of 78 (average parity 5) Landrace × Yorkshire sows. Results showed that swelling capacity was higher in the RS diet than in the CON or FSF diet. Meanwhile, the 48 h cumulative gas production and the final asymptotic gas volume after in vitro fermentation of gestation diets showed an increased trend (p = 0.07, p = 0.09, respectively) in the RS diet versus the CON or FSF diets. While the sows' litter size, body weight, backfat or weaning-to-estrus interval were not affected (p > 0.05) by the three treatments during gestation, the RS group showed a decline in stillbirth number (p < 0.05) and stillbirth rate (p < 0.01) relative to the other two groups. Meanwhile, the proportion of standing was lower while the sow's serum concentrations of PYY (peptide YY) and GLP-1 (glucagon-like peptide-1) were higher (p < 0.05) on day 70 of gestation in the RS group than in the CON or FSF group. Compared with the CON group, the RS group showed a downward tendency (p = 0.07) in the sows' plasma cortisol concentration on day 70 of gestation. A comparison of oxidative and antioxidative indicators revealed an increase in the sows' serum FRAP (ferric ion reducing antioxidant power) (p < 0.05) and a decrease of protein carbonyl (p < 0.05) on day 109 of gestation in the RS or FSF group versus the CON group. Overall, inclusion of 5% RS with greater swelling capacity in the gestation diet contributed to enhancing the postprandial satiety, alleviating the stress status, reducing the abnormal behaviors and thus lowering the stillbirth rate of sows.
RESUMO
The enzymatic properties of four lipases (A, B, C and D) from different strains of Aspergillus niger, were investigated, and a 3-factor mixture design and triangular surface analysis were performed to screen the optimal combi-lipase by observing synergistic effects. Lipases B and D differed in optimal pH, temperature and substrate specificity. A combi-lipase with 31.2% lipase B and 68.8% lipase D (w/w, equal to units of 30.36% and 69.64%) exhibited optimal hydrolytic activity on soybean oil, which exceeded the sum of the combined activities of individual lipases (P < 0.05). Free fatty acid from the hydrolyzed soybean oil indicated that the synergistic effect of the combi-lipase resulted from the different fatty acid specificities of the two lipases. Overall, combi-lipase afforded an effective route for the application of lipase enzymes to animal feeds.
Assuntos
Aspergillus niger/enzimologia , Hidrólise/efeitos dos fármacos , Lipase/farmacologia , Óleo de Soja/química , Ração Animal , Concentração de Íons de Hidrogênio , Lipase/isolamento & purificação , Especificidade por Substrato , TemperaturaRESUMO
Porcine skeletal muscle fibres are classified based on their different physiological and biochemical properties. Muscle fibre phenotype is regulated by several independent signalling pathways, including the mitogen-activated protein kinase (MAPK), nuclear factor of activated T cells (NFAT), myocyte enhancer factor 2 (MEF2) and peroxisome proliferator-activated receptor (PPAR) signalling pathways. MicroRNAs are non-coding small RNAs that regulate many biological processes. However, their function in muscle fibre type regulation remains unclear. The aim of our study was to identify miRNAs that regulate muscle fibre type during porcine growth to help understand the miRNA regulation mechanism of fibre differentiation. We performed Solexa/Illumina deep sequencing for the microRNAome during 3 muscle growth stages (63, 98 and 161 d). In this study, 271 mature miRNAs and 243 pre-miRNAs were identified. We detected 472 novel miRNAs in the muscle samples. Among the mature miRNAs, there are 23 highest expression miRNAs (over 10,000 RPM), account for 85.3% of the total counts of mature miRNAs., including 10 (43.5%) muscle-related miRNAs (ssc-miR-133a-3p, ssc-miR-486, ssc-miR-1, ssc-miR-143-3p, ssc-miR-30a-5p, ssc-miR-181a, ssc-miR-148a-3p, ssc-miR-92a, ssc-miR-21, ssc-miR-126-5p). Particularly, both ssc-miR-1 and ssc-miR-133 belong to the MyomiRs, which control muscle myosin content, myofibre identity and muscle performance. The involvement of these miRNAs in muscle fibre phenotype provides new insight into the mechanism of muscle fibre regulation underlying muscle development. Furthermore, we performed cell transfection experiment. Overexpression/inhibition of ssc-miR-143-3p in porcine skeletal muscle satellite cell induced an/a increase/reduction of the slow muscle fibre gene and protein (MYH7), indicating that miR-143 activity regulated muscle fibre differentiate in skeletal muscle. And it regulate MYH7 through the HDAC4-MEF2 pathway.
Assuntos
MicroRNAs/genética , Fibras Musculares Esqueléticas/fisiologia , Cadeias Pesadas de Miosina/genética , Cadeias Pesadas de Miosina/metabolismo , Animais , Perfilação da Expressão Gênica , Regulação da Expressão Gênica no Desenvolvimento , Sequenciamento de Nucleotídeos em Larga Escala , Masculino , Fibras Musculares Esqueléticas/citologia , Análise de Sequência de RNA , Suínos/crescimento & desenvolvimentoRESUMO
This study was conducted to investigate the effect of dietary protease supplementation on the growth performance, nutrient digestibility, intestinal morphology, digestive enzymes and gene expression in weaned piglets. A total of 300 weaned piglets (21 days of age Duroc × Large White × Landrace; initial BW = 6.27 ± 0.45 kg) were randomly divided into 5 groups. The 5 diets were: 1) positive control diet (PC), 2) negative control diet (NC), and 3) protease supplementations, which were 100, 200, and 300 mg per kg NC diet. Results indicated that final BW, ADG, ADFI, crude protein digestibility, enzyme activities of stomach pepsin, pancreatic amylase and trypsin, plasma total protein, and intestinal villus height were higher for the PC diet and the supplementations of 200 and 300 mg protease per kg NC diet than for the NC diet (P < 0.05). Supplementations of 200 and 300 mg protease per kg NC diet significantly increased the ratio of villus height to crypt depth (VH:CD) of duodenum, jejunum and ileum compared with NC diet (P < 0.05). Feed to gain ratio, diarrhea index, blood urea nitrogen, and diamine oxidase were lower for the PC diet and supplementations of 200 and 300 mg protease per kg NC diet than for the NC diet (P < 0.05). Piglets fed the PC diet had a higher peptide transporter 1 (PepT1) mRNA abundance in duodenum than piglets fed the NC diet (P < 0.05), and supplementations of 100, 200 and 300 mg protease per kg NC diet increased the PepT1 mRNA abundance in duodenum (P < 0.05) comparing with the NC diet. Piglets fed the PC diet had a higher b0,+AT mRNA abundance in jejunum than piglets fed the NC diet (P < 0.05), and supplementations of 200 and 300 mg protease per kg NC diet increased the b0,+AT mRNA abundance in jejunum and ileum comparing with the NC diet (P < 0.05). In summary, dietary protease supplementation increases growth performance in weaned piglets, which may contribute to the improvement of intestinal development, protein digestibility, nutrient transport efficiency, and health status of piglets when fed low digestible protein sources.