Effects of dietary Clostridium butyricum and rumen protected fat on meat quality,oxidative stability,and chemical composition of finishing goats

Background Clostridium butyricum(CB)is a probiotic that can regulate intestinal microbial composition and improve meat quality.Rumen protected fat(RPF)has been shown to increase the dietary energy density and provide essential fatty acids.However,it is still unknown whether dietary supplementation with CB and RPF exerts beneficial effects on growth performance and nutritional value of goat meat.This study aimed to investigate the effects of dietary CB and RPF supplementation on growth performance,meat quality,oxidative stability,and meat nutritional value of finishing goats.Thirty-two goats(initial body weight,20.5±0.82 kg)were used in a completely randomized block design with a 2 RPF supplementation(0 vs.30 g/d)×2 CB supplementation(0 vs.1.0 g/d)factorial treatment arrangement.The experiment included a 14-d adaptation and 70-d data and sample collection period.The goats were fed a diet consisted of 400 g/kg peanut seedling and 600 g/kg corn-based concentrate(dry matter basis).Result Interaction between CB and RPF was rarely observed on the variables measured,except that shear force was reduced(P<0.05)by adding CB or RPF alone or their combination;the increased intramuscular fat(IMF)content with adding RPF was more pronounced(P<0.05)with CB than without CB addition.The pH24h(P=0.009),a*values(P=0.007),total antioxidant capacity(P=0.050),glutathione peroxidase activities(P=0.006),concentrations of 18:3(P<0.001),20:5(P=0.003)and total polyunsaturated fatty acids(P=0.048)were increased,whereas the L*values(P<0.001),shear force(P=0.050)and malondialdehyde content(P=0.044)were decreased by adding CB.Furthermore,CB supplementation increased essential amino acid(P=0.027),flavor amino acid(P=0.010)and total amino acid contents(P=0.024)as well as upregulated the expression of lipoprotein lipase(P=0.034)and peroxisome proliferator-activated receptorγ(PPARγ)(P=0.012),and downregulated the expression of stearoyl-CoA desaturase(SCD)(P=0.034).The RPF supplementation increased dry matter intake(P=0.005),averaged daily gain(trend,P=0.058),hot carcass weight(P=0.046),backfat thickness(P=0.006),concentrations of 16:0(P<0.001)and c9-18:1(P=0.002),and decreased the shear force(P<0.001),isoleucine(P=0.049)and lysine content(P=0.003)of meat.In addition,the expressions of acetyl-CoA carboxylase(P=0.003),fatty acid synthase(P=0.038),SCD(P<0.001)and PPARγ(P=0.022)were upregulated due to RPF supplementation,resulting in higher(P<0.001)content of IMF.Conclusions CB and RPF could be fed to goats for improving the growth performance,carcass traits and meat quality,and promote fat deposition by upregulating the expression of lipogenic genes of Longissimus thoracis muscle.

Supplemental Clostridium butyricum modulates lipid metabolism by reshaping the gut microbiota composition and bile acid profile in IUGR suckling piglets

Background Intrauterine growth restriction(IUGR)can cause lipid disorders in infants and have long-term adverse effects on their growth and development.Clostridium butyricum(C.butyricum),a kind of emerging probiotics,has been reported to effectively attenuate lipid metabolism dysfunctions.Therefore,the objective of this study was to investigate the effects of C.butyricum supplementation on hepatic lipid disorders in IUGR suckling piglets.Methods Sixteen IUGR and eight normal birth weight(NBW)neonatal male piglets were used in this study.From d 3to d 24,in addition to drinking milk,the eight NBW piglets(NBW-CON group,n=8)and eight IUGR piglets(IUGR-CON group,n=8)were given 10 mL sterile saline once a day,while the remaining IUGR piglets(IUGR-CB group,n=8)were orally administered C.butyricum at a dose of 2×108colony-forming units(CFU)/kg body weight(suspended in 10 mL sterile saline)at the same frequency.Results The IUGR-CON piglets exhibited restricted growth,impaired hepatic morphology,disordered lipid metabolism,increased abundance of opportunistic pathogens and altered ileum and liver bile acid(BA)profiles.However,C.butyricum supplementation reshaped the gut microbiota of the IUGR-CB piglets,characterized by a decreased abundance of opportunistic pathogens in the ileum,including Streptococcus and Enterococcus.The decrease in these bile salt hydrolase(BSH)-producing microbes increased the content of conjugated BAs,which could be transported to the liver and function as signaling molecules to activate liver X receptorα(LXRα)and farnesoid X receptor(FXR).This activation effectively accelerated the synthesis and oxidation of fatty acids and down-regulated the total cholesterol level by decreasing the synthesis and promoting the efflux of cholesterol.As a result,the growth performance and morphological structure of the liver improved in the IUGR piglets.Conclusion These results indicate that C.butyricum supplementation in IUGR suckling piglets could decrease the abundance of BSH-producing microbes(Streptococcus and Enterococcus).This decrease altered the ileum and liver BA profiles and consequently activated the expression of hepatic LXRαand FXR.The activation of these two signaling molecules could effectively normalize the lipid metabolism and improve the growth performance of IUGR suckling piglets.

Clostridium butyricum Z-10的生长特性与培养基优化

对Clostridium butyricum Z 10的生长特性研究表明,其最适生长温度为36 ℃,最低和最高生长温度分别为16 ℃和44 ℃;最适起始生长pH值为7.2,最低和最高起始生长 pH值分别为4 6和10 6;最高耐胆酸盐质量浓度为4 g/dL,起始氧化还原电位为-21 mV;丁酸梭状芽孢杆菌对青霉素不敏感,主要代谢产物为丁酸和乙酸.通过对生长因子、氮源的选择性研究,得出采用酵母膏做为生长因子,胰蛋白胨作为氮源有较好的效果.优化培养基试验得出当胰蛋白胨质量浓度为22 g/L,葡萄糖质量浓度为 10 g/L,盐酸半胱氨酸质量浓度为 0 5 g/L时,总菌体浓度可达到3 6×108个/mL.

Effects of dietary supplementation of probiotic,Clostridium butyricum,on growth performance,immune response,intestinal barrier function,and digestive enzyme activity in broiler chickens challenged with Escherichia coli K88

Background： Colibacillosis caused by enterotoxigenic Escherichia coil （E. coil} results in economic losses in the poultry industry. Antibiotics are usually used to control colibacillosis, however, E. coli has varying degrees of resistance to different antibiotics. Therefore the use of probiotics is becoming accepted as an alternative to antibiotics. In this study, we evaluated the effects of Clostfidium butyricum （C. butyficum） on growth performance, immune response, intestinal barrier function, and digestive enzyme activity in broiler chickens challenged with Eschefichia coli （E. coil） K88. Methods： The chickens were randomly divided into four treatment groups for 28 days. Negative control treatment （NC） consisted of birds fed a basal diet without E. coil K88 challenge and positive control treatment （PC） consisted of birds fed a basal diet and challenged with E. coil K88. C. buO/ricum probiotic treatment （CB） consisted of birds fed a diet containing 2 x 107 cfu C. buO/ricum/kg of diet and challenged with E. coil K88. Colistin sulfate antibiotic treatment （CS） consisted of birds fed a diet containing 20 mg colistin sulfate/kg of diet and challenged with E. coil K88. Results： The body weight （BW） and average day gain （ADG） in the broilers of CB group were higher （P 〈 0.05） than the broilers in the PC group overall except the ADG in the 14-21 d post-challenge. The birds in CB treatment had higher （P 〈 0.05） concentration of tumor necrosis factor-a （TNF-a） at 3 and 7 d post-challenge, and higher （P 〈 0.05） concentration of interleukin-4 （IL-4） at 14 d post-challenge than those in the PC treatment group. The concentration of serum endotoxin in CB birds was lower （P 〈 0.05） at 21 d post-challenge, and the concentrations of serum diamine oxidase in CB birds were lower （P 〈 0.05） at 14 and 21 d post-challenge than in PC birds. Birds in CB treatment group had higher （P 〈 0.05） jejunum villi height than those in PC, NC, or CS treatment at 7, 14, and 21 d post-challenge. In comparison to PC birds, the CB birds had lower （P 〈 0.05） jejunum crypt depth during the whole experiment. The birds in CB or CS treatment group had higher （P 〈 0.05） activities of amylase and protease at 3, 7, and 14 d post-challenge, and higher （P 〈 0.05） activity of lipase at 3, 7 d post-challenge than PC birds.

Effects of dietary Clostridium butyricum supplementation on growth performance,intestinal development,and immune response of weaned piglets challenged with lipopolysaccharide

Background: Weanling pigs, with immature immune system and physiological function, usually experience postweaning diarrhea. This study determined the effects of dietary Clostridium butyricum supplementation on growth performance, diarrhea, and immunity of weaned pigs challenged with lipopolysaccharide(LPS).Methods: In Experiment(Exp.) 1,144 weaned piglets were weaned at 21 d and randomly assigned to six groups,with six replicates per group and four pigs per replicate, receiving a control diet(CON) or diet supplemented with antibiotics(AB) or C. butyricum(CB)(0.1%, 0.2%, 0.4%, or 0.8%), respectively. All diets in Exp. 1 were a highly digestible basal diet, with 3,000 mg/kg zinc oxide supplied in the first 2 wk only. In Exp. 2, 180 piglets were weaned at 21 d and randomly assigned to five groups, with six replicates per group and six pigs per replicate, receiving CON, AB, or CB(0.2%, 0.4%, or 0.6%) diets. The digestibility of diets was lower than those in Exp. 1, and did not include zinc oxide. At 36 d of Exp. 2, 12 piglets were selected from each of the CON and 0.4% CB groups, six piglets were intraperitoneally injected with LPS(50 μg/kg body weight) and the other six piglets with normal saline;animals were killed at 4 h after injection to collect blood, intestine, and digesta samples for biochemical analysis.Results: In Exp. 1, CB and AB diets had no effect on growth performance of piglets. In Exp. 2, 0.4% CB decreased feed-gain ratio(P < 0.1), diarrhea score(P < 0.05), and increased duodenal, jejunal, and ileal villus height and jejunal villus height/crypt depth(P < 0.05). The 0.4% CB decreased the plasma tumor necrosis factor(TNF) α(P < 0.05) but increased ileal mucosa IL-10 and TLR2 mRNA expression(P < 0.05). Furthermore, 0.4% CB altered the microbial profile, with Bacillus and Ruminococcaceae UGG-003 at genus level and Lactobacillus casei and Parasutterella secunda at species level were higher than CON in colonic content(P < 0.05).Conclusions: Dietary C. butyricum supplementation had positive effects on growth of weaned piglets with less digestible diets. There was a tendency to reduce the feed-gain ratio, which could reduce feed costs in pig production. Moreover, C. butyricum decreased post-weaning diarrhea by improving the intestinal morphology,intestinal microflora profile, and immune function.

产氢菌Clostridium butyricum P22的分离鉴定及其产氢特性研究

从连续生物制氢反应器中分离得到高效产氢细菌P22,利用微生物自动鉴定仪并结合菌株16S rDNA序列分析,将其鉴定为Clostridium butyricum P22。同时还进一步研究了碳源、氮源、初始pH值和培养温度对菌株P22产氢活性的影响。结果表明:该菌株能利用多种碳源氮源产氢,当以葡萄糖作为产氢底物时,其最大产氢量和产氢速率分别为为301.7 mL/g-glucose(2.42 mol/mol-glucose)、31.1 mL(/g-glucose.h)。发酵液的初始pH和培养温度严重影响产氢菌活性,初始pH6.47、35℃培养为该菌种较为适宜的产氢条件。利用Arrehnius方程以葡萄糖为底物拟合得到该菌株产氢反应的活化能为70.7 kcal/mol。经能量分析,Clostridium butyricum P22产氢过程中葡萄糖能量回收率为20.6%。该菌株在生物制氢中具有很好的应用发展前景。

Effects of dietary supplementation with Clostridium butyricum on the growth performance and humoral immune response in Miichthys miiuy

The effects of dietary supplementation with Clostridium butyricum on growth performance and humoral immune response in Miichthys miiuy were evaluated. One hundred and fifty Miichthys miiuy weighing approximately 200-260 g were divided into five groups and reared in 15 tanks with closed circuiting culture system. The animals were fed 5 diets： basal diet only （control） or supplemented of the basal diet with C. butyricum at doses of 10^3 （CB1）, 10^5 （CB2）, 10^7 （CB3） or 10^9 （CB4） CFU/g. Compared with the control, the serum phenoloxidase activity was significantly increased by the supplementation （P〈0.05）, acid phosphatases activity was increased significantly （P〈0.05） at the doses of 10^9 CFU/g. Serum lysozyme activity peaked at dose of 10^7 CFU/g and in the skin mucus at dose of 10^9 CFU/g. Immunoglobulin M level in the serum and skin mucus was increased except at dose of 10^3 CFU/g （P〈0.05）. The growth at the dose of 10^9 CFU/g was higher than that of the control （P〈0.05）. It is concluded that supplementation of C. butyricum can mediate the humoral immune responses and improve the growth performance in Miichthys miiuy.

Therapeutic effects of Clostridium butyricum on experimental colitis induced by oxazolone in rats

AIM： To evaluate the therapeutic effects of a probiotic supplement （Clostridium butyricum, CGMCC0313 ） in a chemically-induced rat model of experimental colitis. METHODS： An experimental ulcerative colitis model was established by rectal injection of oxazolone into the colon of 40 Wistar rats randomly divided into four groups. The positive control group was sacrificed 3 d after colitis onset. The remaining groups were fed daily with either 2 mL of C. butyricum （2.3 × 10^11 CFU/L）, 2 mL of mesalamine （100 g/L）, or 1 mL of sodium butyrate （50 mmol/L） for 21 d. The animals＇ body weight, behavior, and bowel movements were recorded weekly. After sacrifice, visual and microscopic observations of pathological changes of colon tissue were made, body weight and wet colon mass index were measured and recorded, and serum levels of interleukin-23 （IL-23） and TNF-α were measured using ELISA. Expression of calcitonin gene-related peptide in colon tissue was measured by RT-PCR. Finally, changes in rat intestinal microflora status were measured in all groups.lowered the serum levels of both IL-23 and tumor necrosis factor-α （TNF-α） with similar or even better efficiency than that of mesalamine or sodium butyrate. The rat intestinal flora appeared to recover more quickly in the group treated with C. butyricum than in the mesalamine and sodium butyrate groups. Finally, we found that the expression level of calcitonin gene related peptide was elevated in colon tissue in the sodium butyrate treated group but not in the C. butyricum or mesalamine treated groups, indicating a sensitization of colon following sodium butyrate treatment. CONCLUSION： In our experimental colitis model, treatment with C. butyricum CGMCC0313, a probiotic supplement, is at least as efficient as treatment with mesalamine.

Effect of Clostridium butyricum on fecal flora in Helicobacter pylori eradication therapy

AIM： To investigate the effect of probiotic bacterium, Clostridium butyricum MIYAIRI 588 strain （CBM） on the changes of the fecal flora in Helicobacter pylori （H pylon） treatment. METHODS： Thirty-five patients with gastric or duodenal ulcers positive for H pylori were randomized either to 1 wk amoxicillin, clarithromycin, lansoprazole （Group 1） or to the same regimen supplemented with CBM 7 d ahead of the triple therapy （Group 2）. Stool samples were collected before and 2, 4, 7, 15, and 22 d after the starting eradication therapy, and were examined intestinal flora. Patients were required to keep a diary record of their condition. RESULTS： Obligate anaerobes decreased significantly on d 2, 4, 8 and 15 in Group 1. On the other hand, they did not decrease significantly in Group 2. The Escherichia coli was dominant bacterium in Enterobacteriaceae, but that was replaced by other species such as Klebsiella and Enterobacter after eradication in Group 1. The change was suppressed in Group 2. Abdominal symptoms were less frequent in Group 2 than in Group 1. CONCLUSION： The combined use of CBM reduced the changes in the intestinal flora and decreased the incidence of gastrointestinal side effects.

Effects of the Dietary Probiotic Clostridium butyricum on Intestine Digestive and Metabolic Capacities, SCFA Content and Body Composition in Marsupenaeus japonicus

A 56-day feeding trial was performed to evaluate the effects of dietary probiotic Clostridium butyricum(CB) on intestine digestive and metabolic capacities, intestine short-chain fatty acids(SCFA) content and body composition of kuruma shrimp Marsupenaeus japonicus. Shrimp s were randomly allocated into 9 tanks, 30 each, and f ed with diets containing different levels of C. butyricum(1×10~9 cfug^(-1)): 0 mgg^(-1) feed(Control), 100 mgg^(-1) feed(CB-100), 200 mgg^(-1) feed(CB-200), while each level was triplicated. The results indicated that compared with the control group, the intestine pepsin(Pep) activity and 5-hydroxytryptamine(5-HT) concentration of two C. butyricum group s were both increased. Amylase(AM Y) and lipase(LPS) activities wer e only induced in CB-200 group. Alanine aminotransferase(ALT) and aspartate aminotransferase(A ST) activities of two C. butyricum group s showed no significant change. The α-amylase(AM Y) gene expression was induced in CB-200 group, and tryp sin gene expression of two C. butyricum treated group s were both induced. Intestine SCFA content and body composition analysis showed that the contents of propionic acid, butyric acid and the crude protein of two C. butyricum group s were all higher than those of control. These results revealed that C. butyricum can modulate intestine digestive and metabolic capacities, improve intestine SCFA content and body crude protein content in M. japonicus.

Batch and fed-batch production of butyric acid by Clostridium butyricum ZJUCB

The production of butyric acid by Clostridium butyricum ZJUCB at various pH values was investigated. In order to study the effect of pH on cell growth, butyric acid biosynthesis and reducing sugar consumption, different cultivation pH values ranging from 6.0 to 7.5 were evaluated in 5-L bioreactor. In controlled pH batch fermentation, the optimum pH for cell growth and butyric acid production was 6.5 with a cell yield of 3.65 g/L and butyric acid yield of 12.25 g/L. Based on these results, this study then compared batch and fed-batch fermentation of butyric acid production at pH 6.5. Maximum value (16.74 g/L) of butyric acid concentration was obtained in fed-batch fermentation compared to 12.25 g/L in batch fermentation. It was concluded that culti- vation under fed-batch fermentation mode could enhance butyric acid production significantly (P<0.01) by C. butyricum ZJUCB.

Clostridium butyricum蔗糖磷酸化酶的酶学性质及其功能研究

本研究克隆表达丁酸梭菌Clostridium butyricum DSM10702菌株的蔗糖磷酸化酶基因,并探究重组酶的酶学性质和转糖苷功能。从丁酸梭菌C.butyricum DSM10702菌株中克隆蔗糖磷酸化酶基因cbsp,构建重组表达质粒pQE30-cbsp并在大肠杆菌Escherichia coli XL1-blue中诱导表达;将重组蛋白经镍柱纯化后,进行酶学性质和转糖苷功能研究。实验结果表明,以蔗糖为底物,重组酶Cbsp的最适温度为45℃、最适pH为7.0、V_(max)和K_(m)分别为(957±23.29)μmol·mg^(-1)·min^(-1)和(62.4±4.749) mmol·L^(-1)。当以葡萄糖-1-磷酸(G-1-P)为糖基供体时,重组酶Cbsp对于多数单糖、糖醇类、(-)-儿茶素、己基-β-D-吡喃葡萄糖苷和L-抗坏血酸表现出转糖苷活性。以上研究结果可丰富蔗糖磷酸化酶相关数据库,为其应用研究提供参考。

Potential protective effects of Clostridium butyricum on experimental gastric ulcers in mice

AIM: To investigate the effects of Clostridium butyricum(C. butyricum) on experimental gastric ulcers(GUs) induced by alcohol, restraint cold stress, or pyloric ligation in mice, respectively.METHODS: One hundred and twenty mice were randomly allocated into three types of gastric ulcer models(n = 40 each), induced by alcohol, restraint cold stress, or pyloric ligation. In each GU model, 40 mice were allocated into four groups(n = 10 each): the sham control group; model group(GU induction without pretreatment); C. butyricum group(GU induction with C. butyricum pretreatment); and Omeprazole group(GU induction with Omeprazole pretreatment). Theeffects of C. butyricum were evaluated by examining the histological changes in the gastric mucosal erosion area, the activities of superoxide dismutase(SOD) and catalase(CAT), the level of malondialdehyde(MDA), and the contents of interleukin(IL)-1b, tumor necrosis factor(TNF)-α, leukotriene B4(LTB4) and 6-keto-PGF-1α(degradation product of PGI2) in the gastric tissue.RESULTS: Our data showed that C. butyricum significantly reduced the gastric mucosal injury area and ameliorated the pathological conditions of the gastric mucosa. C. butyricum not only minimized the decreases in activity of SOD and CAT, but also reduced the level of MDA in all three GU models used in this study. The accumulation of IL1-b, TNF-α and LBT4 decreased, while 6-keto-PGF-1α increased with pretreatment by C. butyricum in all three GU models.CONCLUSION: Our data demonstrated the protective effects of pretreatment with C. butyricum on antioxidation and anti-inflammation in different types of GU models in mice. Further studies are needed to explore its potential clinical benefits.

Clostridium butyricum alleviates intestinal low-grade inflammation in TNBS-induced irritable bowel syndrome in mice by regulating functional status of lamina propria dendritic cells

BACKGROUND Irritable bowel syndrome (IBS) is one of the most common functional gastroenterological diseases characterized by abnormal visceral sensitivity and lowgrade inflammation. The role of Clostridium butyricum (C. butyricum) in reducing intestinal low-grade inflammation via immune pathways has been well defined. However, the detailed mechanisms of the effects of C. butyricum on intestinal mucosal immunity, especially on immune cells of the lamina propria, remain unclear. Dendritic cells (DCs), which are important immune cells, secrete proinflammatory cytokines (IL-1β, IL-6, and others) and express T cell immunoglobulin and mucin domain-3 (TIM3), promoting proliferation and activation of DCs, and mediating Th1 and Th17 inflammatory responses. AIM To investigate the role of DCs in the development of IBS in a rat model and to understand the regulation of DCs after C. butyricum intervention. METHODS An IBS animal model was established using C57BL/6 mice, and C. butyricum was continuously administered via the intragastric route to simulate different intestinal immune states. Intestinal visceral hypersensitivity and histopathology were assessed using the abdominal withdrawal reflex (AWR) test and hematoxylin & eosin (H&E) staining, respectively. The expression of proinflammatory cytokines (IL-1β and IL-6) and TIM3 was analyzed by Western blot analysis and real-time PCR. Flow cytometry was applied to analyze the quantity, function, and membrane molecule TIM3 of the lamina propria dendritic cells (LPDCs). The regulatory effect of C. butyricum was verified in bone marrowderived dendritic cells by in vitro experiments. RESULTS The secretion of proinflammatory cytokines (IL-1β and IL-6) in mice with IBS was significantly increased compared with that of the control group, which suggested that the intestinal mucosa in mice with IBS was in a low-grade inflammatory state. The expression of CD11C+CD80+ and CD11c+TIM3+ in intestinal LPDCs in mice with IBS increased significantly. Meanwhile, the cytokines (IL-1β and IL-6) were significantly reduced after the intervention with probiotic C. butyricum. The amount and function of LPDCs and the TIM3 on the surface of the LPDCs were decreased with the alleviation of the intestinal inflammatory response. CONCLUSION The results suggest that C. butyricum regulates the amount and functional status of LPDCs in the intestinal mucosa of mice with IBS, and therefore modulates the local immune response in the intestine.

SCFA Profile of Rice RS Fermentation by Colonic Microbiota, <i>Clostridium butyricum</i>BCC B2571, and <i>Eubacterium rectale</i>DSM 17629

Resistant starch type 3 (RS3) produced from high amylose food sources through retrogradation or enzymatic process is known to have physiological function as dietary fiber. Fermentation of RS3 by colonic microorganisms produced SCFA (acetate, propionate, and butyrate), maintained the health of colon, balance of gut microbiota, preventing inflammatory bowel diseases (IBD) and colon cancer. RS3 in this study was produced from IR-42 and Inpari-16 broken rice by enzymatic treatment (combination of amylase-pullulanase). The Resistant Starch was fermented for 12 and 24 h by colonic microbiota (extracted from healthy human subject), Clostiridium butyricum BCC-B2571, or Eubacterium rectale DSM 17629. SCFA produced was analyzed by gas chromatography. Treatment by amylase-pullulanase combination was advantageous to increase their RS3 content. The result showed that after enzymatic process, the RS3 content of IR-42 (41.13%) was not significantly different (p 0.05) from that of Inpari-16 (37.70%). High concentration of acetate (82.5 mM) and propionate (7.5 mM) were produced by colonic microbiota after 12 h fermentation and best concentration of butyrate (6.8 mM) was produced by colonic microbiota after 24 h fermentation. It is clear that utilization of colonic microbiota rather than single strain was better in the production of SCFA.

Dietary supplementation with Clostridium butyricum improves growth performance of broilers by regulating intestinal microbiota and mucosal epithelial cells

Clostridium butyricum has been widely considered an antibiotic substitute in recent years.It can promote growth performance,improve the immune response and enhance the intestinal barrier function of the host.In the present study,1-d-old Arbor Acres(AA)broilers were fed C.butyricum(1×109 cfu/kg)for 28 d.The transcriptomic characteristics of epithelial cells of the cecal mucosa were determined by RNA-sequence,and the cecal microbiota composition was explored by 16 S ribosomal RNA gene sequencing.The changes in the intestinal mucosa of broilers were then analyzed by tissue staining.Gene Ontology(GO)annotations identified substance transport and processes and pathways that might participate in intestinal development and cell viability.Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analysis revealed that the differentially expressed genes are involved in numerous pathways related to amino acid and vitamin metabolism and antioxidant and defensive functions,among others.The relative expression of some genes associated with intestinal barrier function(claudins 2,15,19,and 23,tight junction proteins 1,2,and 3 and mucin 1)was significantly increased in the treatment group(P<0.05 or P<0.01).Moreover,the proportion of Firmicutes was higher in the C.butyricum-treated group,whereas the proportion of Proteobacteria was higher in the control group.At the genus level,the relative abundances of Butyricicoccus and Lactobacillus,among other bacteria,were increased after C.butyricum supplementation.The tissue staining analysis showed that the cecal mucosa of broilers was significantly ameliorated after the addition of C.butyricum(P<0.05 or P<0.01).These results showed that dietary supplementation with C.butyricum can enhance the antioxidant capacity,mucosal barrier function,and stabilize the cecal microbiota,resulting in improving the growth performance.

电子穿梭体对菌株Clostridium butyricum LQ25异化铁还原性质影响

【目的】在异化铁还原细菌培养体系中,通过外加电子穿梭体,分析电子穿梭体种类与浓度对细菌异化铁还原性质的影响。【方法】以一株发酵型异化铁还原细菌Clostridium butyricum LQ25为研究对象,设置水溶性介体蒽醌-2-磺酸钠和核黄素作为外加电子穿梭体。【结果】在氢氧化铁为电子受体、葡萄糖为电子供体培养条件下,不同浓度蒽醌-2-磺酸钠和核黄素对菌株LQ25异化铁还原效率影响具有显著性差异。外加蒽醌-2-磺酸钠浓度为0.5 mmol/L时,菌株累积产生Fe(Ⅱ)浓度最高,为12.95±0.08 mg/L,相比对照组提高88%。核黄素浓度为100mg/L时,菌株累积产生Fe(Ⅱ)浓度是11.06±0.04mg/L,相比对照组提高61%。外加电子穿梭体能够改变菌株LQ25发酵产物中丁酸和乙酸浓度,提高乙酸相对含量。【结论】蒽醌-2-磺酸钠和核黄素作为外加电子穿梭体能显著促进细菌异化铁还原效率,为揭示发酵型异化铁还原细菌胞外电子传递机制提供实验支持。

丁酸梭菌对花鲈幼鱼生长性能、免疫消化及肠道菌群的影响

【目的】探究丁酸梭菌(Clostridium butyricum)对花鲈(Lateolabrax maculatus)幼鱼生长性能,消化、免疫和抗氧化能力及肠道菌群的影响,为丁酸梭菌在花鲈鱼养殖中的应用提供理论依据。【方法】以花鲈幼鱼为试验材料设饲料中丁酸梭菌添加量为0(D0,对照)、0.25%(D1)、0.50%(D2)、1.00%(D3)、2.00%(D4)和4.00%(D5)6组,投喂花鲈幼鱼56 d,测定其生长性能、肠道消化酶活性、血清生化和免疫指标及肠道形态指标,分析肠道菌群组成。【结果】丁酸梭菌可显著降低饲料系数(P<0.05,下同),显著提高花鲈幼鱼的终末平均体质量,D3、D4和D5组的增重率及特定生长率均显著高于D0组。与D0组相比,D4组肠道糜蛋白酶和脂肪酶活性均显著提高,血清总胆固醇、甘油三酯含量和谷草转氨酶、谷丙转氨酶活性均显著降低。D4组溶菌酶、碱性磷酸酶和总一氧化氮合酶活性均显著高于D0组,D3组酚氧化酶活性显著高于D0组;与D0组相比,D3和D5组过氧化氢酶活性及总抗氧化能力均显著提高,D4组超氧化物歧化酶活性显著提高,丙二醛含量显著降低。D2、D3和D5组花鲈幼鱼肠道肌层厚及绒毛高度均显著高于D0组。D3组花鲈幼鱼肠道菌群OTU数量和Chao1指数均显著高于D0组,各试验组花鲈幼鱼肠道菌群Simpson指数和Shannon指数均显著高于D0组;在门分类水平上,丁酸梭菌降低了变形菌门和厚壁菌门相对丰度,提高了放线菌门、浮霉菌门、酸杆菌门、绿弯菌门、拟杆菌门和芽单胞菌门相对丰度;在属分类水平上,丁酸梭菌降低了劳尔氏菌属、肠弧菌属和弧菌属有害菌相对丰度,提高了节杆菌属和KD4-96_unclassified有益菌相对丰度。【结论】饲料中添加丁酸梭菌能显著提高花鲈幼鱼生长性能,提高消化、免疫和抗氧化能力,调节肠道菌群结构组成及其相对丰度,提高肠道菌群多样性,以2.00%添加量效果最好。

3种不同添加物对凡纳滨对虾生长、非特异免疫和抗病力的影响

为评估3种不同添加物对凡纳滨对虾(Litopenaeus vannamei)生长性能、非特异免疫反应及抗病力的影响,在基础饲料中分别添加了丁酸梭菌CBG01(Clostridium butyricum CBG01)活菌(CB组)、3%聚β-羟基丁酸酯(PHB组)和1%丁酸钠(BS组)来投喂对虾,对照组投喂基础饲料,养殖6周后测量对虾生长情况、检测血清非特异免疫指标和肝胰腺免疫相关基因表达水平,进行副溶血弧菌(Vibrio parahaemolyticus)攻毒实验。研究表明,CB组和PHB组对虾末体质量和特定生长率显著高于对照组(P<0.05),CB组最高,BS组与对照组差异不显著(P>0.05)。3个处理组对虾成活率和饲料效率均显著高于对照组(P<0.05)。3个处理组对虾血清中溶菌酶、过氧化物酶、超氧化物歧化酶活性以及总抗氧化能力显著高于对照组(P<0.05),CB组对虾血清中碱性磷酸酶、酸性磷酸酶、酚氧化酶活性显著高于对照组(P<0.05),PHB组对虾血清中碱性磷酸酶、酸性磷酸酶、总一氧化氮合酶活性显著高于对照组(P<0.05),BS组对虾血清中酸性磷酸酶活性显著高于对照组(P<0.05)。与对照组相比,CB组对虾肝胰腺中所有免疫相关基因(SOD、LZM、proPO、LGBP、HSP70、Imd、Toll、Relish、TOR、4E-BP、eIF4E1α、eIF4E2)相对表达量均显著上调(P<0.05),而PHB组的Toll基因以及BS组的Imd、Toll、Relish、eIF4E2基因相对表达量与对照组无显著差异(P>0.05)。副溶血弧菌攻毒实验表明,3个处理组对虾的累积死亡率均显著低于对照组(P<0.05)。研究结果表明,饲料中添加丁酸梭菌、PHB和丁酸钠均可不同程度地提高凡纳滨对虾的生长性能和非特异免疫能力,能够显著提高对虾对副溶血弧菌感染的抵抗力。总体比较,添加丁酸梭菌和PHB的作用效果基本相当,在一定程度上要优于添加丁酸钠。在饲料生产加工过程中,在难以添加丁酸梭菌活菌的情况下,建议适当添加聚β-羟基丁酸酯来替代丁酸梭菌。

饲粮复合添加丁酸梭菌和葡萄糖氧化酶对生长肉兔生长性能、屠宰性能、免疫和抗氧化功能以及盲肠微生物区系的影响

本试验旨在研究饲粮复合添加丁酸梭菌和葡萄糖氧化酶对生长肉兔生长性能、屠宰性能、免疫和抗氧化功能以及盲肠微生物区系的影响。选取250只体重相近的35日龄断奶伊拉商品肉兔,随机分为5组,每组50个重复,每个重复1只兔。对照组饲喂基础饲粮,抗生素组饲喂在基础饲粮中添加有效含量为75 mg/kg金霉素和20 mg/kg恩拉霉素的饲粮,试验组分别饲喂在基础饲粮中添加300 g/t丁酸梭菌(试验1组)、200 g/t丁酸梭菌+200 g/t葡萄糖氧化酶(试验2组)和300 g/t丁酸梭菌+200 g/t葡萄糖氧化酶(试验3组)的饲粮。预试期7 d,正试期35 d。结果表明:1)与对照组相比,试验2组和试验3组肉兔平均日增重(ADG)显著提高(P<0.05),料重比(F/G)显著降低(P<0.05),且这2个组与抗生素组相比均无显著差异(P>0.05);试验2组死亡率显著低于对照组(P<0.05),且腹泻频率和死亡率与抗生素组无显著差异(P>0.05)。由此可见,试验2组替抗效果最好。2)各组肉兔屠宰性能和免疫器官指数均无显著差异(P>0.05)。3)与对照组相比,试验2组肉兔血清促炎细胞因子白细胞介素-1β(IL-1β)含量有降低趋势(P=0.085),血清白细胞介素-6(IL-6)含量显著降低(P<0.05)。4)与对照组相比,试验2组肉兔肝脏谷胱甘肽过氧化物酶(GSH-Px)活性显著提高(P<0.05)。5)与对照组相比,试验2组肉兔盲肠微生物群落丰富度和进化多样性有提高的趋势(0.05<P<0.10)。6)与对照组相比,试验2组肉兔盲肠微生物特有扩增子序列变异(ASV)数提高了42.79%,2组间显著差异的标志物种可能是ASV_19729(P<0.05),属于厚壁菌门的梭菌属。综上所述,肉兔饲粮中复合添加200 g/t丁酸梭菌和200 g/t葡萄糖氧化酶能够提高其生长性能和成活率,降低机体炎症反应,增强肝脏抗氧化能力,改善盲肠微生物群落结构,替代抗生素效果较好。