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.

Impacts of dietary supplementation of bamboo vinegar and charcoal powder on growth performance,intestinal morphology,and gut microflora of large-scale loach Paramisgurnus dabryanus

Although accumulating data demonstrate that dietary supplementation of bamboo vinegar and charcoal powder(BVC)can significantly strengthen the immunity and boost the growth of domestic animals,its application potency still awaits verification in commercial fish species such as the large-scale loach Paramisgurnus dabryanus.The impacts of 90-day dietary supplementation of 1%and 2%BVC on survival and growth performance of the loach,and on the intestinal morphological characteristics and gut microflora were analyzed.Our data show that the large-scale loach supplied with BVC at the experimental doses had significant higher survival rates and better growth performance(indicated by greater weight gain(1.13–1.14 times),higher specific growth rate(1.04 times),and lower feed conversion ratio(0.88–0.89 times))compared to that of the control(P<0.05).Histological examination revealed significant longer villus(3.22–5.54 times),deeper crypt(1.77–1.87 times),and thicker muscle(1.59–3.17 times)in the intestines of large-scale loach fed with BVC(P<0.05).Furthermore,we found that the gut microflora consisted of significantly fewer proportions of potential pathogenic bacterial species(Aeromonas veronii and Escherichia coli),but significantly greater proportions of beneficial microbes(Lactococus raffinolactis and Faecalibacterium prausnitzii).Therefore,dietary intake of BVC can promote intestinal tract development and optimize gut microflora,by which the survival and growth of large-scale loach may be improved.

Study on the interaction between β-carotene and gut microflora using an in vitro fermentation model

β-Carotene,a typical non-oxygenated carotenoid,is the most efficient source of retinol(VA).The low bio-availability ofβ-carotene lead to large accumulation in colon;however,the relationship betweenβ-carotene and gut microflora remains unclear.This study intends to explore the interaction betweenβ-carotene and gut microflora using an in vitro fermentation model.After 24 h fermentation,the degradation rate ofβ-carotene was(64.28±6.23)%,which was 1.46 times that of the group without gut microflora.Meanwhile,the production of VA was nearly 2 times that of the group without gut microflora,indicating that the gut microflora can metabolizeβ-carotene into VA.β-Carotene also influences the production of short-chain fatty acids(SCFAs),the production of total SCFAs in 0.5 mg/mLβ-carotene(BCM)group was(44.00±1.16)mmol/L,which was 2.26 times that of the blank control(BLK)group.Among them,the production of acetic acid in BCM group was(19.06±0.82)mmol/L,which was 2.64 time that of the BLK group.Furthermore,β-carotene significantly affected the structure and composition of gut microflora,increasing the abundance of Roseburia,Parasutterella and Lachnospiraceae,and decreasing the abundance of Dialister,Collinsella and Enterobacter(P<0.05).This study provides a new way to understand howβ-carotene works in human body with gut microflora.

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.

Research progress on adaptive modifications of the gut microflora and regulation of host glucose and lipid metabolism by cold stimulation

The gut microflora is a combination of all microbes in intestine and their microenvironment,and its change can sensitively reflect the relevant response of the body to external environment and remarkably affect body's metabolism as well.Recent studies have found that cold exposure affects the body's gut microflora,which can lead to changes in the body's metabolism of glucose and lipid.This review summarizes recent research on the effects of cold exposure on gut microbes and metabolism of glucose and lipid,aiming to provide some new ideas on the approaches and measures for the prevention and treatment of diabetes and obesity.

Mucosal bacterial microflora and mucus layer thickness in adolescents with inflammatory bowel disease

AIM:To assess the mucosa-associated bacterial microflora and mucus layer in adolescents with inflammatory bowel disease(IBD) .METHODS:Sixty-one adolescents(mean age 15 years,SD ± 4.13) were included in the study.Intestinal biopsies from inflamed and non-inflamed mucosa of IBD patients and from controls with functional abdominal pain were cultured under aerobic and anaerobic conditions.The number of microbes belonging to the same group was calculated per weight of collected tissue.The mucus thickness in frozen samples was measured under a fluorescent microscope.RESULTS:The ratios of different bacterial groups in inflamed and non-inflamed mucosa of IBD patients and controls were specific for particular diseases.Streptococcus spp.were predominant in the inflamed mucosa of Crohn's disease(CD) patients(80% of all bacteria) ,and Lactobacillus spp.were predominant in ulcerative colitis patients(90%) .The differences were statistically significant(P = 0.01-0.001) .Lower number of bifidobacteria was observed in the whole IBD group.A relation was also found between clinical and endoscopic severity and decreased numbers of Lactobacillus and,to a lesser extent,of Streptococcus in biopsies from CD patients.The mucus layer in the inflamed sites was significantly thinner as compared to controls(P = 0.0033) and to non-inflamed areas in IBD patients(P = 0.031) .CONCLUSION:The significantly thinner mucosa of IBD patients showed a predominance of some aerobes specific for particular diseases,their numbers decreased in relation to higher clinical and endoscopic activity of the disease.

Effects of Salvia miltiorrhiza on intestinal microflora in rats with ischemia/reperfusion liver injury

BACKGROUND: Hepatic ischemia/reperfusion injury may induce intestinal microflora imbalance. Salvia miltiorrhiza is effective in promoting blood circulation and counteracting peroxidation in tissues. The aim of the present study was to determine the effects of Salvia miltiorrhiza on intestinal mi- croflora, endotoxemia, and bacterial translocation in rats with hepatic I/R injury. METHODS: Sprague-Dawley rats in specific pathogen free grade were divided into 3 groups: group I(n =6) for sham operation: groups ( n = 7) for liver ische- mia for 20 minutes and reperfusion for 22 hours. Group was also pretreated with 4 ml/day of Salvia miltiorrhiza solu- tion (250 mg/kg) by daily gavage for 7 days. The levels of serum alanine aminotransferase (ALT), aspartate amino- transferase (AST), malondialdehyde ( MDA) and supero- xide dismutase ( SOD ) in liver tissues, serum endotoxin, intestinal bacterial counts, intestinal mucosal histology and bacterial translocation were studied. RESULTS: The levels of ALT, AST, plasma endotoxin and MDA in liver tissues were decreased more markedly in group (57.57 ± 18.08 U/L, 147.57 ±40.84 U/L, 0.42 ± 0.144 EU/ml and 0. 52 ±0.19 nmol/mg-prot respectively) in group 295.9±216.92 U/L, 0.80± 0.262 EU/ml and 0.72±0.12 nmol/mg-prot; P <0.05-0.01 respectively). Liver SOD activity was increased more sig- nificantly in group (318.47±64.62 U/mg-prot) than in group U/mg-prot, P<0.05). The counts of Bifidobacteria and Bacteroides increased more significantly in group than in group but were similar to those in group I. Bacterial translocation to the kidney in group was 50% (5/10), whereas no bacterial translocation to the kidney occurred in the other two groups (P <0. 01). Ileal mucosal structure was markedly ameliorated in group as compared with group CONCLUSIONS: Salviae miltiorrhiza could partially restore intestinal microflora balance, improve intestinal mucosal integrity, and reduce bacterial translocation and plasma en- dotoxin in rats with hepatic ischemia/reperfusion injury.

Effect of lactobacillus on the gut microflora and barrier function of the rats with abdominal infection

AIM:To investigate the effect of probiotics supplemented by gut on the tight junctions of epithelial cells, barrier function and the microflora of rats with abdominal infection. METHODS: After the model of cecal ligation and perforation established, SD rats were divided into two groups: parenteral nutrition (PN) group and PIM+probiotics (probiotics) group, PN solution was supplemented by neck vein and probiotics was delivered via the jejunostomy tube for five days. Vena cava blood and the homogenated tissue of liver, lung and mesenteric lymph nodes were cultured to determine the bacterial translocation rate (BTR). The ultra-structure of epithelial tight junctions and microvilli of the gut were observed by electron microscopy; occluding expression was measured by indirect-immune fluorescence method; anaerobic bacterial growth by anaerobic culture and DNA fingerprint of bacterial colonies of the feces by PCR. RESULTS: The quantity of lactobacteria and bifydobacteria in probiotics group was higher than that of PN group. The profiles of DNA fingerprint expression in probiotics group were similar to that in the normal group, a new 16S rDNA sequence appeared in the profile in PN group. The occludin expression, the integrality of the gut epithelial tight junction and microvilli in probiotics group were improved as compared with PN group. The BTR and endotoxin in blood were reduced more significantly in probiotics group as compared with PN group. CONCLUSION: The probiotics could improve the gut microflora disturbance, increase occludin expression, maintain the gut epithelial tight junction and decrease the bacterial translocations rate.

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.

Intestinal microflora in rats with ischemia/reperfusion liver injury

Objectives: To investigate the intestinal microflora status related to ischemia/reperfusion (I/R) liver injury and explore the possible mechanism. Methods: Specific pathogen free grade Sprague-Dawley rats were randomized into three groups: Control group (n=8), sham group (n=6) and I/R group (n=10). Rats in the control group did not receive any treatment, rats in the I/R group were subjected to 20 min of liver ischemia, and rats in the sham group were only subjected to sham operation. Twenty-two hours later, the rats were sacrificed and liver enzymes and malondialdehyde (MDA), superoxide dismutase (SOD), serum endotoxin,intestinal bacterial counts, intestinal mucosal histology, bacterial translocation to mesenteric lymph nodes, liver, spleen, and kidney were studied. Results: Ischemia/reperfusion increased liver enzymes, MDA, decreased SOD, and was associated with plasma endotoxin elevation in the I/R group campared to those in the sham group. Intestinal Bifidobacteria and Lactobacilli decreased and intestinal Enterobacterium and Enterococcus, bacterial translocation to kidney increased in the I/R group compared to the sham group. Intestinal microvilli were lost, disrupted and the interspace between cells became wider in the I/R group.Conclusion: I/R liver injury may lead to disturbance of intestinal microflora and impairment of intestinal mucosal barrier function,which contributes to endotoxemia and bacterial translocation to kidney.

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.

Soil Physico-Chemical Properties and Microflora as Influenced by Bispyribac Sodium 10% SC in Transplanted Kharif Rice

The effects of bispyribac sodium 10% SC and butachlor 50% SC on soil physico-chemical properties and microflora in transplanted kharifrice were investigated over two seasons （2010 and 2011 ） Effects of the herbicide on bulk density, water holding capacity, moisture content, soil pH, organic matter content, electrical conductivity, as well as total nitrogen, available phosphorus and available potassium contents were analyzed along with microflora population （total bacteria, actinomycetes and fungi）. No significant changes in soil physico-chemical properties were observed. Herbicide treatments resulted in decreases in microbial counts initially. With the degradation of applied herbicides within a considerable time, the microflora populations even exceeded the initial count at 60 d after application of the herbicide.

Effects of bar-transgenic rice on the intestinal microflora of the mice (<i>Mus musculus</i>)

Microbial molecular ecology approaches were used to the effects of Bar-transgenic rice on Intestinal Micro-flora of the Mice (Mus musculus). Kunming mice (Mus musculus) of 100 SPF-grade (20 g ± 2 g), half of which were male and the other half female, were randomly divided into five groups with four replications per group and five mice per replication to assess the safety of Bar-transgenic rice. Five diets meetinging or exceeding the minimum nutrient requirement were fed for 180 days. After 90 days, parental generation (P) was bred to produce the first filial generation (F1). Each generation was fed for 180 days. On the 180th day, five mice from each group were randomly sampled, and their intestinal contents were collected for DNA isolation. The V3 region of the 16S rDNA was amplified by polymerase chain reaction (PCR) and analyzed via denaturing gradient gel electrophoresis (DGGE). The resulting PCR-DGGE band number (bacterial species) was counted, and the banding patterns were analyzed by calculating the Sorenson’s pairwise similarity coefficients (Cs), an index used to measure bacterial species found among all samples. The sequence analysis of bands was performed to identify the intestinal predominant microflora of the mice. The intergroup Cs values of the samples across all groups did not differ (P > 0.05) from each other. The effect of Bar-transgenic rice on the intestinal microflora of the mice was considered insignificant.

Effects of soybean meal replacement with fermented alfalfa meal on the growth performance,serum antioxidant functions,digestive enzyme activities,and cecal microflora of geese

Fermented forages are important feedstuffs. Bacillus subtilis inoculants are often used to improve the value of forage legume fermentation. The present work was conducted to study the effects of replacing soybean meal with solid-state fermented alfalfa meal（FAM） with B. subtilis ACCC 01746 on growth performance, serum antioxidant and digestive enzyme activities, and cecal microflora in goose. 300 healthy geese with similar body weights were randomly assigned to six treatment groups with five replicates of 10 geese（five males and five females） each. Geese were fed ad libitum for 35 days. Results showed that the geese fed with 4 and 8% FAM exhibited no significant effects on their final body weights（FBW） and average day gain（ADG）（P〉0.05）, whereas 12% or higher FAM caused poor growth of the geese compared with control diet（linear（L）： P〈0.05）. The average daily feed intake（ADFI）（quadratic（Q）： P〈0.05） and feed conversion ratio（FCR）（L： P〈0.05） with 8% or higher supplementation level were higher than those of the control group. The activities of antioxidant enzyme in serum increased, and the level of malondialdehyde（MDA） significantly decreased with increasing dietary FAM level（L： P〈0.05）. However, no significant differences were observed at 8% or lower supplementation level for glutathione peroxidase（GSH-Px） and superoxide dismutase（SOD）（P〉0.05） and at 4% for catalase（CAT） supplementation level compared with the control group. All diets containing FAM increased digestive enzyme activities in geese. However, geese fed diets with 12% FAM supplementation showed the highest trypsin activities in pancreas（Q： P〈0.05）. Supplementation with 12% or higher FAM significantly increased amylase activities in pancreas（L： P〈0.05） and duodenum（L： P〈0.05） compared with the control group. Significant differences were not observed in total anaerobic bacteria between geese fed with FAM and control diets on day 35（P〉0.05）. The numbers of Bifidobacterium and Lactobacillus in the cecum of geese fed with FAM significantly increased（L： P〈0.05）, but no significant effects were found with 4 and 8% FAM supplementation levels compared with the control（P〉0.05）. By contrast, the coliform counts of cecum decreased with increasing inclusion of FAM, but these counts were significantly reduced in geese fed diets with 12% or higher FAM supplementation level（L： P〈0.05）. Collectively, our results indicated that supplementation of the basal geese diet with 8% FAM had no apparent adverse effects on growth performance, serum antioxidant enzyme activities, and digestive parameters and beneficial microbiota.

Effects of dietary forage to concentrate ratio and wildrye length on nutrient intake, digestibility, plasma metabolites, ruminal fermentation and fecal microflora of male Chinese Holstein calves

Twenty-eight male, weaned Chinese Holstein calves（（156.8±33.4） kg） were used to investigate the effects of dietary forage to concentrate ratio（F：C） and forage length on nutrient digestibility, plasma metabolites, ruminal fermentation, and fecal microflora. Animals were randomly allocated to four treatments in a 2×2 factorial arrangement： whole-length forage（WL） with low F：C（50：50）; WL with high F：C（65：35）; short-length forage（SL） with high F：C（65：35）; and SL with low F：C（50：50）. Chinese wildrye was used as the only forage source in this trial. The grass in the SL treatments was chopped using a chaff cutter to achieve small particle size（-50% particles 〉19 mm）. Dry matter intake（DMI） and organic matter（OM） intake was increased by increasing both F：C（P〈0.01） and forage length（FL）（P〈0.05）, while acid detergent fiber（ADF） and neutral detergent fiber（NDF） intakes were only increased by increasing the F：C（P〈0.01）. The digestibility of NDF was increased as the FL increased（P〈0.01）, and it was also affected by interaction between F：C and FL（P〈0.05）. Cholesterol（CHO）（P〈0.01）, leptin（LP）（P〈0.05）, and growth hormone（GH）（P〈0.01） concentrations in plasma were increased as dietary F：C increased. A significant increase in plasma triglyceride（TG）（P〈0.01）, insulin（INS）（P〈0.05）, and GH（P〈0.01） levels was observed with decreasing dietary FL. Ruminal p H values of calves fed with low F：C diets were significantly lower than those in high F：C treatment（P〈0.05）. Increasing the F：C enhanced ruminal acetic acid（P〈0.05） and acetic acid/propionic acid（P〈0.01）. Fecal Lactobacillus content was significantly higher, while Escherichia coli and Salmonella contents were significantly lower in WL and high F：C groups（P〈0.05）. Lower fecal scores（higher diarrhea rate） were observed in calves fed with SL hay compared to WL hay（P〈0.05）. Denatured gradient gel electrophoresis（DGGE） bands and richness index（S） were significantly affected by the interaction between F：C and FL（P〈0.05）, under high F：C, band numbers and richness index from WL group were higher than that from SL group（P〈0.05）, whereas there were no differences between WL andSL groups under low F：C（P〉0.05）. Microflora similarity was 50–73% among the different treatments. It is concluded that the WL with high F：C（65：35） diet is suitable for weaned calves.

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 Winter Cover Crop and Straw Returning on the Functional Diversity of Rhizosphere Microflora in Double-crop Rice Paddies

The functional diversity of rhizosphere microflora which is also known as the “microbial community” is a sensitive indicator of soil quality subject to the type of winter cover crop and straw returning.In order to evaluate the effects of different winter cover crops and returning patterns on the functional diversity of rhizosphere microflora in double-crop rice paddies,we designed five winter cover crops and straw returning combinations to analyze their effects on the functional diversity of rhizosphere microflora in rice paddies: ryegrass (Lolium multiflorumL.)–double-crop rice (Ry),milk vetch (Astragalus sinicus L.)–double-crop rice (Mv),Rape (Brassica napus L.)–Double-crop rice (Ra), Potato (Solanum tuberosum L.)–double-crop rice (Po),and winter fallow–doublecrop rice (CK,the control).In this paper,the average well color development (AWCD) in Biolog-GN plates indicated the capacity for carbon utilization by the rhizosphere microbial community.We analyzed the rhizosphere microbial community functional diversity of the paddy soils with the above five treatments by using the Biolog-GN system.The results showed that applications of winter cover crop and straw returning caused high increases in AWCD compared with CK,and the AWCD values for samples with Po treatment was greater than those with Ry and CK treatments at the early and late rice maturity stages. It was concluded that applications of winter cover crop and straw returning can cause changes in the carbon utilization efficiency of rhizosphere microflora.There were differences in the genetic diversity of the rhizosphere microflora among different treatments at the maturity stage of early and late rice.The richness,Shannon,and McIntosh Index under different winter cover crop and straw returning treatments were significantly different.The highest indexes were observed in the Po treatment and the lowest in the CK at the maturity stage of early and late rice.The richness, Shannon,and McIntosh Index under different treatments ranked in descending order is as follows: Po>Ra>Mv>Ry>CK.Principal Component Analysis (PCA) of substrate reactions were conducted in this research.The results indicated that the pattern of carbon source utilization varied with winter cover crop treatments,and that carbohydrates and amino acids were the main carbon sources of rhizosphere microorganisms.To conclude,the application of winter cover crop and straw returning to paddy fields could significantly increase the carbon source utilization, species richness,and species evenness of rhizosphere microflora in double-crop rice paddies.