Baizhu-Baishao herb pair ameliorates functional constipation and intestinal microflora disorder in rats

Background:In China,Rhizoma atractylodis macrocephalae-Paeonia lactiflora Pall(Biazhu-Baishao,BZBS)is a classic herb pair used to treat intestinal stress syndrome,ulcerative colitis and other diseases.However,the mechanism of BZBS in the treatment of functional constipation(FC)has been little studied and remains unclear.In this study,a behavioral investigation,colon tissue morphology,enzyme-linked immunosorbent assay(Elisa)and intestinal microflora analysis have been used to illuminate the potential mechanism of the effects of BZBS on FC in a rat model.Methods:A FC rat model was constructed and BZBS was given as treatment.Observations and recordings were made of the fecal moisture content,the defecation time of the first black stool,and the rate of intestinal propulsion.Elisa was used to detect the expression levels of substance P(SP),vasoactive intestinal peptide(VIP),5-hydroxytryptamine(5-HT)in the colon.To ascertain the composition of the microbial community,a high throughput 16S ribosomal RNA(16S r RNA)gene sequencing technique was employed.Results:Oral administration of BZBS significantly ameliorated several key excretion parameters,including the time to first black stool defecation,stool water content,and the propulsion rate in the small intestine in FC rats.It increased the expression of SP,VIP and 5-HT in the colon.16S r RNA gene sequencing results showed that BZBS changed the microbial community structure,decreased the Bacteroidetes/Firmicutes ratio,increased the relative abundance of Blautia and Fusicatenibacter,and decreased the relative abundance of Ruminococcus and Roseburia.Conclusions:BZBS effectively alleviates FC and improves dysbacteriosis.

Effects of a Combination of Fu Zi and Rou Gui on Intestinal Neurotransmitters and Microflora in Rats with Slow Transit Constipation

[Objectives] This study was carried out to explore the combined effects of Fu Zi(Radix Aconiti Lateralis Praeparata, the secondary root of perennial herbaceous plant Acontium carmichaeli Dehx. of family Ranunculaceae) and Rou Gui(Cortex Cinnamomi, the bark of Cinnamamunz cassia Presl of family Lauraceae) on intestinal neurotransmitters and microflora in rats with slow transit constipation(STC). [Methods] Experimental rats were given loperamide hydrochloride by gavage to induce STC, and then treated with Fu Zi alone, Rou Gui alone, a combination of Fu Zi and Rou Gui(2:1 w/w), and prucalopride, respectively, for 14 days. Meanwhile, the general condition, the time to first black stool and the rate of intestinal propulsion of rats in each group were observed after STC was induced and after drug treatment, and the pathological changes in rat colon were observed via hematoxylin-eosin(HE) staining, and the levels of colonic 5-hydroxytryptamine(HT), vasoactive intestinal peptide(VIP) and substance P(SP) were detected by ELISA, and the changes in intestinal flora were detected by 16S rRNA Real-time PCR. [Results] Compared with healthy rats, the time to first black stool and the rate of intestinal propulsion, colonic 5-HT and SP levels significantly decreased(p<0.01), while their colonic VIP level significantly increased(p<0.01). Compared with STC rats, the time to first black stool, the rate of intestinal propulsion, colonic 5-HT and SP levels in Fu Zi-Rou Gui(2:1) treated rats and prucalopride treated rats significantly increased(p<0.01), while their colonic VIP level significantly decreased(p<0.01). There was no significant difference in alpha diversity between healthy rats and STC rats. However, analysis on beta diversity revealed that there were differences in microflora structure and composition between them. Compared with healthy rats, the relative abundance of Firmicutes and Proteobacteria in STC rats significantly increased, while that of Bacteroidetes decreased. Compared with STC rats, the relative abundance of Proteobacteria decreased and that of Bacteroidetes and Firmicutes increased in Fu Zi-Rou Gui(2:1) treated rats;the relative abundance of Bacteroidetes and Proteobacteria decreased while that of Firmicutes increased in Fu Zi treated rats;the relative abundance of Proteobacteria decreased while that of Bacteroidetes increased in Rou Gui treated rats;the relative abundance of Firmicutes and Proteobacteria decreased while that of Bacteroidetes increased in prucalopride treated rats. The intestinal flora in rats of all groups was dominated by Lactobacillus spp. and other genera of anaerobic bacteria. Compared with healthy rats, the relative abundance of Lactobacillus spp. and Clostridium spp. in STC rats decreased, while those of Blautia spp. and Ruminococcus spp. and Allobaculum spp. increased. Compared with STC rats, the relative abundance of Lactobacillus spp. in all rats treated with drugs increased. [Conclusions] The combination of Fu Zi and Rou Gui(2:1) can effectively improve intestinal motility in STC rats by regulating intestinal microbial community and the levels of colonic neurotransmitters.

Effect of synbiotics on intestinal microflora and digestive enzyme activities in rats

AIM: To investigate the effect of synbiotics, i.e. probiotics and prebiotics mixture, on the gut microbial ecology and digestive enzyme activities in rats. METHODS: Forty-eight SD rats weighing about 280 g were used in this study. Rats were divided into three groups according to the contents of probiotics and prebiotics mixture in the feed as control, low and high dose groups. The duration of the experiment was 8 wk. RESULTS: Compared with the control group, thefecal Lactobacillus and Bifidobacterium counts were significantly increased and the fecal Coliform organism counts were markedly reduced in the low and high dose groups. Concerning the digestive enzyme activity of jejunum, only lactase activity increased in low dose group. However, significant increase of lipase, lactase, sucrase, and isomaltase activities were observed in high dose group.CONCLUSION: Intake of low and high dosages of probiotics and prebiotics mixture significantly improved the ecosystem of the intestinal tract by increasing the probiotics population and digestive enzyme activities in rats.

Changes in intestinal microflora in rats with acute respiratory distress syndrome

AIM: To implement high-throughput 16S rDNA sequencing to study microbial diversity in the fecal matter of rats with acute lung injury/acute respiratory distress syndrome (ALI/ARDS).

A simple and rapid method for extracting bacterial DNA from intestinal microflora for ERIC-PCR detection

AIM: To develop a simple and convenient method for extracting genomic DNA from intestinal microflora for en- terobacterial repetitive intergenic consensus (ERIC)-PCR detection. METHODS: Five methods of extracting bacterial DNA, including Tris-EDTA buffer, chelex-100, ultrapure water, 2% sodium dodecyl sulfate and 10% Triton-100 with and without sonication, were compared with the commercial fecal DNA extraction kit method, which is considered as the gold standard for DNA extraction. The comparison was based on the yield and purity of DNA and the indexes of the structure and property of micro-organisms that were reflected by ERIC-PCR. RESULTS: The yield and purity of DNA obtained by the chelex method was similar to that obtained with the fecal DNA kit. The ERIC-PCR results obtained for the DNA extracted by the chelex method and those obtained for DNA extracted with the fecal DNA kit were basically the same.CONCLUSION: The chelex method is recommended for ERIC-PCR experiments in view of its simplicity and cost- effectiveness; and it is suitable for extracting total DNA from intestinal micro-organisms, particularly for handling a large number of samples.

Characterizing the composition of intestinal microflora by 16S rRNA gene sequencing

BACKGROUND This study determined the composition and diversity of intestinal microflora in patients with colorectal adenoma(CRA),which may provide precedence for investigating the role of intestinal microflora in the pathogenesis of colorectal tumors,the composition of intestinal microflora closely related to CRA,and further validating the possibility of intestinal flora as a biomarker of CRA.AIM To study the relationship between intestinal microflora and CRA.METHODS This is a prospective control case study from October 2014 to June 2015 involving healthy volunteers and patients with advanced CRA.High-throughput sequencing and bioinformatics analysis were used to investigate the composition and diversity of intestinal microflora in 36 healthy subjects and 49 patients with advanced CRA.Endpoints measured were operational taxonomic units of intestinal flora,as well as their abundance and diversity(αandβtypes).RESULTS In this study,the age,gender,body mass index,as well as location between controls and patients had no significant differences.The mucosa-associated gut microbiota diversity and bacterial distribution in healthy controls and colorectal adenomas were similar.The operational taxonomic unit,abundance,andαandβdiversity were all reduced in patients with CRA compared to controls.At the phylum level,the composition of intestinal microflora was comparable between patients and controls,but the abundance of Proteobacteria was increased,and Firmicutes and Bacteroides were significantly decreased(P<0.05).The increase in Halomonadaceae and Shewanella algae,and reduction in Coprococcus and Bacteroides ovatus,could serve as biomarkers of CRA.High-throughput sequencing confirms the special characteristics and diversity of intestinal microflora in healthy controls and patients with CRA.CONCLUSION The diversity of intestinal microflora was decreased in patients with CRA.An increase in Halomonadaceae and Shewanella algae are markers of CRA.

Effect of herbal medicine Juzentaihoto on hepatic and intestinal heat shock gene expression requires intestinal microflora in mouse

AIM： To evaluate the role of intestinal microflora in the effects of multi-herbal medicine on gene expression in the gut and liver. METHODS： The multi-herbal medicine Juzentaihoto （JTX） was administered to five germ-free mice and regular mice for 2 wk. Among the results of the comprehensive gene chip analysis of the intestine and liver, we featured heat shock proteins （HSPs） 70 and 105 because their gene expression changed only in the presence of microflora. Real-time RT-PCR was performed to confirm the expression levels of these HSP genes. To determine whether JTX acts directly on the HSP genes, sodium arsenite （SA） was used to induce the heat shock proteins directly. To examine the change of the intestinal microflora with administration of JTX, the terminal restriction fragment polymorphism （T-RFLP） method was used. To identify the changed bacteria, DNA sequencing was performed.documented by gene chip and real-time RT-PCR, changed with the administration of JTX in the regular mice but not in the germ-free mice. JTX did not suppress the direct induction of the HSPs by SA. T-RFLP suggested that JTX decreased unculturable bacteria and increased Lactobacillus johnsoni. These data suggested that JTX changed the intestinal microflora which, in turn, changed HSP gene expression.CONCLUSION： Intestinal microflora affects multi-herbal product JTX on the gene expression in the gut and liver.

Bacterial diversity in the intestine of young farmed puffer fish Takifugu rubripes

The aim of the study was to examine the bacterial community associated with the intestinal mucus of young farmed puffer fish Takifugu rubripes. Polymerase chain reaction and partial 16 S r DNA sequencing was performed on DNA from bacteria cultivated on Zobell 2216 E medium. All the isolates were classified into two phyla—Proteobacteria and Firmicutes. Proteobacteria were the dominant, culturable intestinal microbiota(68.3%). At the genus level, V ibrio, Enterobacter, Bacillus, Pseudomonas, Exiguobacterium, Staphylococcus, Acinetobacter, Pseudoalteromonas and S hewanella were isolated from the intestine, with representatives of the genera V ibrio, Enterobacter and Bacillus accounting for 70.7% of the total. This is the first report of Enterobacter, Bacillus, Exiguobacterium and Staphylococcus as part of the intestinal bacterial microflora in T. rubripes. The profile of the culturable bacterial community differed between samples collected from the same tank at 2-month intervals, as indicated by Bray-Curtis and Sorensen indices, and the impact on the intestinal physiology and health of puffer fish requires further investigation.

Effect of Lycopus lucidus Turcz.supplementation on gut microflora and short chain fatty acid composition in Crj:CD-1 mice

We investigated the diversity and composition of microflora in feces of Lycopus lucidus Turcz.-fed mice.In addition,we evaluated the production of major cytokines(Interleukin-6 and-10)which are related to inflammation and fatty acid composition of several tissues.16S ribosomal DNA sequencing-based microbiome taxonomic profiling analysis was performed utilizing the EzBioCloud data base.Male mice fed on L.lucidus showed a significantly reduced number of lactic acid bacteria and coliform in the feces compared with the control group(p<0.05).16S rDNA sequencing analysis of fecal samples showed that L.lucidus supplementation decreased the community of harmful microflora(Enterobacteriaceae including Escherichia coli and Bacteroides sp.)in feces compared with the control group(p<0.05).There were no significant differences in mRNA expression of cytokine IL-6 and IL-10 between the control and L.lucidus fed groups.The fecal fatty acid composition in the L.lucidus group had percentages of 4:0,6:0,8:0 and 10:0 in the intestine but those short chain fatty acids were not detected in the control group.Our results showed that L.lucidus supplementation influenced gut environment by decreasing harmful microflora and increased the percentages of several short fatty acids.

Effects of Spent Mushroom Substrate of Pleurotus eryngii as Fermentation Bed Padding on Growth Performance, Intestinal Microflora and Immunity of Weaning Piglets

[Objective] The paper was to examine the effects of spent mushroom substrate of Pleurotus eryngii as fermentation bed padding on growth performance, intestinal microflora and immunity of weaning pigs. [Method] A total of 120 weaning piglets （DurocxLandracexYorkshire） with the average initial body weight of （8.0±0.5） kg were allocated to five dietary treatments in a randomized complete block design for 42 d, each of which was replicated three times with eight piglets per replicate （ half male and half female）. The padding for control group was 50% sawdust ＋50% rice husk; experimental group I, 100% spent mushroom substrate; experimental group II, 15% sawdust ＋15% rice husk ＋70% spent mushroom sub- strate; experimental group III, 25% sawdust ＋25% rice husk ＋50% spent mushroom substrate; and experimental group IV, 35% sawdust ＋35% rice husk ＋30% spent substrate. [Result] Except for experimental group IV, the other three experimental groups had higher average daily gain compared to the control group （P〈0.05）. The average daily feed intake in experimental group I increased obviously compared to the control group（P〈0.05）. Except for experimental group I, the diarrhea rate of weaning piglets in experimental groups II, III and IV significantly decreased compared to the control group（P〈0.05）. The number of lactobacilli and bifidobacteria in colon and cecum in experimental groups I, II and III increased distinctly （P〈 0.05）, while the number of Escherichia coli and Salmonella decreased remarkably compared to the control group （P〈0.05）. The positive rates of T and B lymphocytes in peripheral blood of weaning piglets in four experimental groups were significantly higher than that in control group at 21 and 42 d post weaning （P〈0.05）. The IgA content of intestinal mucous in piglets was significantly improved in experimental groups II and III （P〈0.05）. [Conclusion] It enhances the production performance when improving immunity and reducing diarrhea rate of piglets by using spent mushroom substrate of P. eryngii as the fermentation bed padding. Experimental group III （25% sawdust ＋25% rice husk ＋50% spent mushroom substrate） is the optimal proportion of spent mushroom substrate.

Development of an Efficient Method for Extracting Total DNA of Intestinal Microflora

[ Objective] The aim was to develop a fast and effective DNA extraction method of intestinal microflora, a modified method of chloroform extraction, and to provide the basis for quantitative and qualitative detection. [ Method] Through the improvement of conventional DNA extraction method, a rapid and efficient DNA extraction method was developed. Compared with the real-time PCR result of control sample and the result of QIAamp DNA Stool Mini kit, the developed method was verified. [ Result] The DNA yield of the developed method was 100 times as much as that of QIAamp DNA Stool Mini kit. And the real-time PCR result showed that the efficiency of DNA extraction of the developed method was higher than that of the QIAamp DNA Stool Mini kit. [ Conclusion] This modified method is inexpensive, efficient and rapid, and it is suitable for large quantities of feces samples.

Effects of Shuanghuanglian Propolis Oral Liquid on Growth Performance and Intestinal Microflora of Broilers

[ Objective] The paper was to investigate the effects of adding different levels of Shuanghuanglian propelis oral liquid in drinking water on growth performance and intestinal microflora of broilers. [ Method] Ninety one-day-old broilers were randomly divided into three groups （group I, group 1I and group III）： group I was the control group, and groups II and III were added with 0.5 and 1.0 mL/L Shuanghuanglian prepolis oral liquid in drinking water, respec- tively. [ Result] Compared with broilers in group I, the average daily gains of broilers in group II and group III increased significantly （ P 〈 0.05 ） ; the feed gain ratio of broilers in group III was significandy lower than that in group I （ P 〈 0.05 ） ; the number of intestinal LactobaciUus spp. in group II and group III in- creased significantly （ P 〈 0.05 ） ; the number of Salmonella spp. and Escherichia coli in group lII decreased significantly （ P 〈 0.05 ） ; the number of Bgrdobacterium spp. increased significantly （ P 〈 0.05 ）. [ Conclusion] Adding 0.5 and 1.0 mL/L Shuanghuanglian propolis oral liquid in drinking water could improve growth performance and intestinal micreflora of broilers, and the dose of 1.0 mL/L is recommended in clinical application.

Research on Mechanism and Application of DUAN-NAI-AN on Controlling Weaned Piglet Diarrhea

[ Objective] The study aimed to explore the function mechanism of microbial ecological agent DUAN-NAI-AN on controlling weaned piglet diarrhea. E Method] DUAN-NAI-AN was given to the weaned piglets by oral administration and its effects on diarrhea rate, growth performance, in- testinal microflora, pH values of different parts of the intestine and intestinal mucosal morphology were observed. [ Result] DUAN-NAI-AN could im- prove the growth performance of weaned piglets by increasing the growth rate and phase weight gain, showing as higher livability, average daily gain and daily feed intake as well as lower diarrhea rate with 19.24% decline by contrast to the control. In addition, DUAN-NAI-AN could keep the intestinal mucosal mechanical barrier and normal digestive function by strengthening intestinal resistance and thus enhance the intestinal mucosal im- mune level. It could also decrease the proportion of potential pathogens and their toxicity effects inside the intestine to help the piglets transfer to and then maintain the dynamic balance of intestinal microflora in a very short period, and it also showed a trend of decreasing the pH value of piglet in- testinal contents. [ Conclusion] DUAN-NAI-AN is an effective microbial ecological agent for the control of weaned piglet diarrhea.

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.

Probiotics in hepatology

The paper provides a basic review of intestinal microflora and its importance in liver diseases. The intestinal microflora has many important functions, above all to maintain the microbial barrier against established as well as potential pathogens. Furthermore, it influences the motility and perfusion of the intestinal wall, stimu- lates the intestinal immune system and therefore also the so-called common mucosal immune system, reducing bacterial translocation and producing vitamins. Immune homeostasis at mucosal level results from a controlled response to intestinal luminal antigens. In liver cirrhosis, there are many changes in its function, mostly an increase in bacterial overgrowth and translocation. In this review, probiotics and their indications in hepatology are generally discussed. According to recent knowledge, these preparations are indicated in clinical practice only for cases of hepatic encephalopathy. Probiotics are able to decrease the permeability of the intestinal wall, and decrease bacterial translocation and endotoxemia in animal models as well as in clinical studies, which is extremely important in the prevention of complications of liver cirrhosis and infection after liver transplantation. Probiotics could limit oxidative and inflammatory liver damage and, in some situations, improve the histological state, and thus non-alcoholic steatohepatitis could be considered as another possible indication.

Protective effect of probiotics on intestinal barrier function in malnourished rats after liver transplantation

BACKGROUND: Most patients waiting for liver transplantation have end-stage liver diseases with malnutrition, which is prone to induce intestinal barrier dysfunction after liver transplantation. We aimed to study the effect of probiotics on intestinal barrier function in malnourished rats following liver transplantation with long-term antibiotics. METHODS: Twelve Lewis rats were selected as donors. Twelve BN rats, which served as recipients, were subjected to malnutrition by semi-starvation for 4-5 weeks. They were randomly divided into two groups: a control group which received phosphate-buffered saline and a probiotics group which received Bifidobacterium and Lactobacillus. All recipients were injected with intramuscular imipenem and subcutaneous cyclosporine A. Furthermore, six normal BN rats without any drugs or operations served as a normal group. Eight days after operation, all rats were sacrificed for examination of the following parameters: serum levels of endotoxin and TNF-α, bacterial translocation, intestinal microflora, ileocecal sIgA, lymphocyte numbers, and phenotypes (CD4, CD8, αβTCR, γδTCR)ofPeyer’spatches. RESULTS: In recipients subjected to malnutrition, weight decreased by 20% and they survived until 8 days after operation. Compared with the normal group, all recipients on postoperative day 8 showed increased levels of serum endotoxin and TNF-α as well as increased counts oftranslocated bacteria. Meanwhile, there were decreases in counts of Bifidobacterium and Lactobacillus in the ileocecum, sIgA concentration, and lymphocytes of Peyer’s patches. Moreover, partial alteration in lymphocyte phenotypes was evidenced by elevated ratios of CD8 + and γδTCR + lymphocytes. In contrast, compared to the control group, supplementation with probiotics reduced the levels of serum endotoxin, TNF-α and bacterial translocation, increased the counts of Bifidobacterium and Lactobacillus, the concentration of sIgA and lymphocytes of Peyer’s patches, and also slightly restored the alteration of lymphocyte phenotypes. CONCLUSION: Supplementation with probiotics including Bifidobac-terium and Lactobacillus promoted partial restoration of intestinal microflora and improved intestinal barrier function in malnourished rats after liver transplantation with long-term use of antibiotics.

Antibiotics and probiotics in treatment of inflammatory bowel disease

Many experimental and clinical observations suggest that intestinal microflora plays a potential role in the pathogenesis of inflammatory bowel disease （IBD）. Manipulation of the luminal content using antibiotics or probiotics represents a potentially effective therapeutic option. The available studies do not support the use of antibiotics in ulcerative colitis （UC）. Antibiotics are effective in treating septic complications of Crohn＇s disease （CD） but their use as a primary therapy is more controversial, although this approach is frequently and successfully adopted in clinical practice.There is evidence that probiotic therapy may be effective in the prevention and treatment of mild to moderate UC. In contrast, a lack of successful study data at present precludes the widespread use of probiotics in the treatment of CD. Both antibiotics and probiotics appear to play a beneficial role in the treatment and prevention of pouchitis and further trials are warranted to fully quantify their clinical efficacy.

Single or combined effects of dietary arabinoxylan-oligosaccharide and inulin on growth performance,gut microbiota,and immune response in Pacific white shrimp Litopenaeus vannamei

The single and synergistic effects of dietary arabinoxylan-oligosaccharide(AXOS)and inulin at different doses(2,4,and 8 mg/g diet)on survival,growth performance,gut microbiota,and immune response in Pacific white shrimp Litopenaeus vannamei were assessed.Singular application of either inulin or AXOS at doses of 4 mg/g diet showed the most stimulatory effects on the growth rate and gene expression levels of chitinase,cathepsin L,chymotrypsin,and extracellular signal-regulated kinase(ERK)in each single prebiotic feeding trial.Compared with single prebiotic treatments,simultaneous application of AXOS and inulin at 4 mg/g diet enhanced remarkably the growth parameters of shrimp and the expression of related genes(chitinase,cathepsin L,chymotrypsin,ERK,myeloid differentiation factor 88,and phenoloxidase)after 8-week feeding(P<0.05).Additionally,gut microbiota analysis indicated the dietary supplementation with combined prebiotics increased significantly the bacterial community richness and relative abundance of Bacillus Pseudomonas Bacteriovorax,and Lactobacillus,and reduced the abundance of Vibrio Rhodococcus,and Photobacterium in the digestive tract of L vannamei.Compared with the single prebiotic treatment and the control,combined prebiotics supplementation boosted notably the survival rate and expression levels of immune-related genes in shrimp infected with Vibrio alginolyticus or white spot syndrome virus.Therefore,simultaneous application of AXOS and inulin shall have a great potential of dietary supplement in the culture of L.vannamei.

Eubacterium limosum ameliorates experimental colitis and metabolite of microbe attenuates colonic inflammatory action with increase of mucosal integrity

AIM： To examine the effect of Eubacteriuro Iiroosuro （E.Iiroosuro） on colonic epithelial cell line in vitro, and to evaluate the effect of E.limosum on experimental colitis.METHODS： E.Iimosum was inoculated anaerobically and its metabolites were obtained. The growth stimulatory effect of the E.limosum metabolites on T84 cells was evaluated by SUDH activity, and the anti-inflammatory effect by IL-6 production. The change in mRNA of toll like receptor 4 （TLR4） was evaluated by real time PCR. Colitis was induced by feeding BALB/C mice with 2.0% dextran sodium sulfate. These mice received either 5% lyophilized E.lirnosum （n=7） or control diet （n=7）. Seven days after colitis induction, clinical and histological scores, colon length, and cecal organic acid levels were determined.RESULTS： The E.Iimosum produced butyrate, acetate, propionate, and lactate at 0.25, 1.0, 0.025 and 0.07 retool/L, respectively in medium. At this concentration, each acid had no growth stimulating activity on T84 cells; however, when these acids were mixed together at the above levels, it showed significantly high activity than control. Except for lactate, these acids significantly attenuated IL-6 production at just 0.1 mmol/L. In addition, under TNF-α stimulation, butyrate attenuated the production of TLR4 mRNA. The treatment with E.limosum significantly attenuated clinical and histological scores of colitis with an increase of cecal butyrate levels, compared with the control group.CONCLUSION： E.limosum can ameliorate experimental colonic inflammation. In part, the metabolite of E.lirnosurn, butyrate, increases mucosal integrity and shows anti-inflammatory action modulation of mucosal defense system via TLR4.

A Robust Noninvasive Approach to Study Gut Microbiota Structure of Amphibian Tadpoles by Feces

The 16S rDNA amplicon high-throughput sequencing technique provides a robust and inexpensive approach to detect the gut microbiota of amphibians. Since different experimental protocols generate technical biases in drawing the gut microbiota profiles, the integrative analysis of gut microbiota produced by different studies must be performed with circumspection. In this study, we compared the efficacy of two DNA extraction methods （i.e., a phenol-chloroform method and TIANamp Stool DNA Kit） in describing intestinal and fecal bacterial communities of transplanted Asiatic toad （Bufo gargarizans） tadpoles. In terms of the DNA extraction quality （i.e., DNA purity and yield rate） and the consistency in between fecal and intestinal microbiota structures （i.e., a and 13 diversity indices）, the phenol-chloroform method was more robust than this commercial stool kit in profiling gut microbiota of tadpoles with feces.