Abstract:Being an important component to the health of the host, the intestinal flora can participate in the immune regulation of the body, promote the development of the body's immune system, and maintain normal immune function. At the same time, the immune system has regulatory and restrictive effects on the intestinal flora. This paper reviews the species of intestinal flora, factors affecting the changes in intestinal flora, the mechanism of interaction between intestinal microbiota and pathogens, and the role in host infection and immune response. This paper will also provide new insights into research on the mechanism of intestinal flora participating in the immune response.

Abstract:The chicken gastrointestinal tract (GIT) harbors a complex microbial community. The intestinal microbiota is closely related to both the intestinal and overall health of the host. In order to get insights into the composition and function of chicken gut microbiota (intestinal microbiome) we provide an overview of the chicken gastrointestinal microbiota by focusing on the establishment and development, spatial distribution in all intestinal compartments and physiological significance. We foresee advances in strategies to isolate and effectively utilize the functional bacteria in chicken intestinal tract and reasonably manage/modulate the intestinal microbe-host interactions to enhance feed efficiency and improve gut health.

Abstract:[Objective] This study was to explore the effect of biochar-based fertilizer application on the microbiological mechanism of the quality of tobacco-planting yellow soil. [Methods] In 2016, 3 different treatments were designed:no fertilizer (NF), conventional fertilizer (CF) and biochar-based fertilizer (BF). Illumina miseq sequencing was applied to analyze soil bacterial and fungal communities. [Results] Compared with NF and CF treatments, the yield of flue-cured tobacco was increased, the contents of soil alkaline hydrolyzed N and available P were increased significantly in the BF treatment. In the BF treatment, the number of operational taxonomic units (OTUs, 1592), abundance and diversity indexes of soil bacterial community were the highest. However, the number of OTUs (280), abundance and diversity indexes of soil fungal community were the lowest. The relative abundance of Bacteroidetes (1.50%) was increased in the BF treatment, whereas Latescibacteria (0.11%) was decreased. There were significant differences in the relative abundance of Bacteroidetes and Latescibacteria between BF and NF treatments. In addition, BF significantly decreased the relative abundance of Zygomycota (2.18%). Compared with the NF treatment, the relative abundance of Chytridiomycota (0.01%) was significantly decreased in the BF treatment. BF changed the fungal community at the genus level and significantly decreased the relative abundance of Fusarium (5.83%). Compared with the NF treatment, the relative abundance of Mortierella (2.11%), Stachybotrys (0.90%) and Alternaria (0.01%), was decreased significantly in the BF treatment. The most important soil factor causing the change of bacterial community structure was pH, and the most important soil factor causing the change of fungal community structure was available P. [Conclusion] Biochar-based fertilizers cause changes in soil biochemical environment, lead to changes in community structure, diversity of soil bacteria and fungi and then optimize soil ecology.

Abstract:[Objective] To facilitate the localization of proteins in budding yeast, we developed yeast reporter strains expressing fusion proteins that localize to specific subcellular structures. [Methods] The yeast expression vectors expressed organelle-specific proteins tagged with much brighter variant red fluorescent protein (RedStar) at their carboxy terminal were constructed and transferred into yeast cells to generate the reporter strain collection. To this end, the yeast codon that optimized coding sequence of RedStar were in-frame inserted immediately preceding the stop codon of each ORF that controlled by their endogenous promoters. Each strain was verified by PCR and sequencing, and then analyzed by fluorescence microscopy. Co-localization of DNA-binding dyes (DAPI), Mitotracker (mitochondria) and SipA from Salmonella enteritidis in the corresponding reporter strain were also performed. [Results] The yeast strain collection containing indicators for the actin, endoplasmic reticulum, nuclei, mitochondria, peroxisomes, lipids, endosomes, and the Golgi apparatus were constructed successfully. [Conclusion] Our studies provide a fundamental tool for observing the dynamic changes of organelles and identifying the localization of unknown proteins in yeast.

Abstract:[Objective] The aim of this study was to screen a strain with high alginate lyase activity and optimize the culture conditions of the strain. [Methods] Using sodium alginate as the sole carbon source, we screened and isolated microorganisms from the coastal soil of Zhangzhou, Fujian province, and a high-yield strain of alginate lyase was obtained. According to the morphological, physiological and biochemical characteristics and 16S rDNA sequence analysis, the target strain was identified. Then the enzyme production conditions were optimized by single factor and orthogonal test. [Results] Four strains with transparent circle to colony diameter ratio (D/d)>3 were obtained by cetylpyridine staining. DNS method was used to determine the activity of alginate lyase in the fermentation broth of four strains. The enzyme activity of strain SH-1 was the highest, reaching 315.5 U/mL. Based on morphological, physiological, biochemical and 16S rDNA sequencing identification, it was named Microbulbifer sp. SH-1. Through single factor and orthogonal test optimization, the optimum enzyme production medium was determined as follows (g/L):sodium alginate 10, NaCl 5, (NH₄)₂SO₄ 5, MgSO₄ 0.2, K₂HPO₄ 1 and FeSO₄ 0.02. Further optimization of culture conditions showed that SH-1 strain could be inoculated into 50 mL medium at initial pH 7.5 and 32℃ with 1% inoculation. The maximum enzyme activity of SH-1 strain could reach 757.9 U/mL under 240 r/min for 24-h shaking, 2.4 times higher than that before optimization. [Conclusion] The optimized conditions for SH-1 production provides reference for large-scale preparation of alginate lyase and further research.

Abstract:[Objective] To study the effect of Chlamydia muridarum on the ulcerative colitis induced by dextran sulfate sodium (DSS) in mice. [Methods] In total 15 female C57BL/6J mice were randomly divided into 3 groups:control group, DSS group, experimental group (CM+DSS group). The mice of CM+DSS group were administrated with 2×10⁵ IFU Chlamydia muridarum by gavage. After 28 days, the mice of the DSS group and CM+DSS group drank 2% DSS water for 5 days, then they were monitored for body weight and disease activity index, the colon length of all mice, and the inflammatory damage of intestine tissue. [Results] The model of ulcerative colitis induced by DSS was well established with the body weight loss of colitis mice, increased disease activity index, short colon length, and severe inflammatory reaction of colon tissue. Compared with the DSS group, CM+DSS group recovered largely, including the body weight loss, disease activity index, colon length, and colon inflammatory damage. [Conclusion] Chlamydia muridarum could ameliorate ulcerative colitis induced by DSS.

Abstract:[Objective] Porcine circovirus type 2 (PCV2) is one of the major etiological agents of porcine circovirus-associated diseases (PCVAD) and could cause severe economic loss to the swine industry worldwide. However, the mechanism of pathogenesis, epidemiology and molecular genetic evolution are not fully understood. Samples obtained from East China between 2010 and 2015 were detected for PCV2. In addition, 42 of 158 PCV2 positive samples were used for the whole genome amplification and sequencing to explore the genetic relationship, molecule epidemiology and genetic evolution of PCV2. [Methods] All of 384 field samples from the clinical diseased pigs between 2010 and 2015 in Jiangsu, Anhui and Zhejiang provinces of China, were screened by specific PCR to detect PCV2. The whole genome of PCV2 positive samples selected randomly was sequenced and analyzed. [Results] PCV2 is distributed widely among swine populations in East China and 158 clinical samples were positive for PCV2, indicating that the prevalence of PCV2 infection in pigs was about 41.15%. The phylogenetic tree constructed by the neighbor-joining method based on the whole genome or ORF2 gene was highly similar. The nucleotide homology and amino acid homology between the ORF2 gene of 42 isolates and 11 reference strains was between 87% to 100% and 82.5% to 100%, respectively. Phylogenetic analysis of the ORF2 genome showed that 53 strains were presented in 4 sub-genotypes (PCV2a, PCV2b, PCV2c and PCV2d). Based on comparing the nucleotides and amino acid sequences of ORF2, 42 isolates obtained in this study shared 89.6% to 100% and 86.8% to 100% identity, respectively. Genetic evolution analysis indicated that 42 isolates could be divided into three genotypes as PCV2a, PCV2b and PCV2d (69.05%), and PCV2b did not appear in Anhui and Zhejiang. Among the three genotypes, PCV2d was the predominant genotype circulating in the swine population of three provinces. Comparisons of the amino acid sequence of Cap protein revealed that 42 isolates in this study have a low variation, four major hypervariable genome regions and special substitutions among the different PCV2 genotypes. There were some aa changes in the reported antibody epitope regions of the Cap protein. However, whether those mutations could result in antigenic variations or change the pathogenicity of PCV2 and the vaccine effect should be explored further. [Conclusion] The results demonstrated that genetic evolution of PCV2 was relatively stable and PCV2b genotype was dominant in the porcine population in East China from 2010 to 2015. According to amino acid alignment data of Cap protein, different substitutions were observed in different genotypes and some representative substitutions exist in each genotype. This study provides a reference for investigating the genetic relationship, genetic evolution and pathogenesis of PCV2 in East China.

Abstract:[Objective] Vibrio parahaemolyticus is a common foodborne pathogen in aquatic products. The formation of biofilm is essential for the survival and spread of Vibrio parahaemolyticus. The aim of this study was to evaluate the structural diversity of biofilms formed by 44 strains of Vibrio parahaemolyticus isolated under clinical and environmental conditions. [Methods] This study proposed a high-throughput method based on confocal laser scanning microscopy (CLSM), using 96-well microtiter plates compatible with high-resolution imaging, combined with structural analysis software ISA-2 to study biofilm formation and structure, the biofilm structure parameters (Biovolume, Average thickness, Biofilmroughness) formed by 22 clinical strains and 22 environmental strains were analyzed. [Results] CLSM images showed that 44 strains of Vibrio parahaemolyticus could form 3D structure after 48 h incubation, and further analyzed the similarities and differences of biofilm formation between clinically derived strains and environmentally derived strains, showed that the coefficient of variation in the clinical strains were smaller than that of the environmental strains, and the strains BF with both tdh and trh virulence factors had the smallest variability. Agglomerative hierarchical clustering (AHC) showed that biofilm can be divided into dense and smooth surface (39%), mottled and rough surface (27%) and loose and uneven surface (34%), the clinical strain formed a biofilm which is easy to form dense and smooth surface, mottled and rough surface, whereas the environmental strain formed a biofilm which is easy to dense and smooth surface and loose and uneven surface. [Conclusion] This study provides an in-depth understanding of the biofilm structure of Vibrio parahaemolyticus, it provides theoretical support for different prevention and control measures for Vibrio parahaemolyticus biofilms from clinical and environmental sources.

Abstract:Biogenetic coalbed methane is a significant composition in Erlian and Hailar basins. However, little information is available how biogenetic coalbed methane is generated. [Objective] To evaluate the methanogenic potential of volatile fatty acids by the coalbed-methane production water microbiome, and to observe the shift of microbial community structure involved. [Methods] Simulation experiments were performed by inoculating coalbed-methane production water from four coalbed methane wells of Erlian and Hailar basins. Acetate, propionate and butyrate were selected as substrates. Methane production and substrate utilization were measured over time. The microbial communities of coalbed methane production water and enrichment cultures after incubation were analyzed through high-throughput sequencing of 16S rRNA gene. [Results] With the exception of H303 coalbed water without propionate degradation, all coalbed water microbiomes had the ability to degrade acetate, propionate and butyrate under methanogenic degradation. The maximum specific methane production rate, substrate conversion ratio and lag-phase period exhibited great difference in different volatile fatty acid group. The maximum specific methane production rate and substrate conversation ratio was highest in the butyrate group and the lag-phase was the shortest with acetate addition. High-throughput sequencing analysis reveals that the archaeal community structure of enrichment cultures was significantly correlated with the source of coalbed water. The predominant archaeal groups of Erlian basin were hydrogenotrophic Methanocalculus (13.5%-63.4%) and Methanosarcina (7.9%-51.3%). The predominant archaea of Hailar basin were hydrogenotrophic Methanobacterium (24.3%-57.4%) and Methanosarcina (29.6%-66.5%). The enriched bacterial community structure is significantly correlated with substrates, sulfate-reducing Desulfovibrio (12.0%-41.0%), syntrophic propionate-degrading Syntrophobacter (39.63%-75.45%) and syntrophic butyrate-degrading Syntrophomonas (8.5%-21.9%) were enriched in acetate, propionate and butyrate group, respectively. [Conclusion] The coalbed-methane production water microbiome possessed potential of methanogenic volatile fatty acid degradation, among which acetate was probably fermented by methanogens, propionate and butyrate are consumed by syntrophic bacteria Syntrophobacter and Syntrophomonas respectively. This result will contribute to the development research of microbial enhanced methane recovery from coalbed.

Abstract:[Objective] Screening biocontrol strains for controlling sugarcane red rot caused by Colletotrichum falcatum Went. [Methods] The endophytic bacteria isolated from sugarcane were used as target bacteria. The sugarcane red rot pathogenic fungus Colletotrichum falcatum Went. was used as an indicator. Strains with strong inhibitory effect on the pathogen were screened by confrontation culture. Antibacterial activity of the metabolites was determined by agar diffusion method. Then, the broad-spectrum analysis of the antimicrobial activity of highly effective strains with good antagonistic effect was carried out and identified. Finally, the high-efficiency strain YC89 was identified through morphological, physiological and biochemical characteristics in combination with 16S rDNA and gyrA sequence analysis. [Results] Five strains of antagonistic bacteria with bacteriostatic bands greater than 1.60 cm were screened by initial screening, among which X22, W2 and YC89 were up to 1.87 cm. The five endophytic bacteria obtained from the initial screening were rescreened. Strains YC89, H1, X22, W2 and YT93 had a inhibition rate above 75% against sugarcane red rot, among which YC89 had the best inhibition rate of 78%. The culture solution, supernatant, filtrate and crude protein extract of strain YC89 showed strong antagonistic activities to the hyphal growth of Colletotrichum falcatum Went. In addition, strain YC89 also had good activity against 7 plant pathogenic fungi. Strain YC89 was identified as Bacillus velezensis. [Conclusion] Strain YC89 has good biocontrol effect on sugarcane red rot pathogen, and has great application potential in the biological control of sugarcane red rot.

Abstract:[Objective] Due to their unique antibacterial activity, silver nanoparticles (AgNPs) have attracted great attention recently. And it is said that AgNPs are extensively antibacterial agents which have strong inhibition and toxic effects on dozens of pathogenic microorganisms. Compared with traditional synthetic methods, biosynthesis way has the advantages of mild reaction conditions and environmental-friendliness and is becoming a hot research topic. [Methods] In this study, we have used the cell-free extracts of fungal Mariannaea sp. HJ to synthesize AgNPs. The conditions for synthesis and the antibacterial activity were also investigated. [Results] Study has shown that the cell-free extracts of 350 mg/L, AgNO₃ 5 mmol/L and pH 7.0 were the optimal synthesis conditions for this reaction. Transmission Electron Microscope (TEM) images showed that the as-synthesized AgNPs were mainly spherical and pseudo spherical with good dispersibility. Diffraction of x-rays (XRD) analysis indicated that the crystal structure of AgNPs were face-centered cubic. The Fourier Transform Infrared Spectrometer (FTIR) analysis results suggested that the hydroxyl, carboxyl and other functional groups in the extracts might have played a functional role in the reduction or stabilization process. Besides, it has been verified that the AgNPs synthesized under the optimal experimental conditions had good antibacterial activity against both Gram-negative bacteria Escherichia coli BL21 and Gram-positive bacteria Arthrobacter sp. W1. [Conclusion] In brief, the cell-free extracts of Mariannaea sp. HJ could greenly synthesize spherical AgNPs with uniform size and good dispersion. The synthesized AgNPs might have potential research value in antibacterial advances.

Abstract:[Objective] To screen and identify resistant strains of Salmonella phage in the process of lysis, the differences of biological characteristics and the pathogenicity between resistant strains and the host strains were studied, to provide theoretical basis for solving the problem of presence of resistant bacteria in the phage therapy. [Methods] Salmonella phage-resistant bacteria were screened by secondary infection method and double-layer plate method. The differences between host strain ATCC 13076 and its phage-resistant strain R3 were compared by biological characteristics and virulence genes detection. The virulence of the bacteria was compared using mice challenge test and cell adhesion test. [Results] The growth curve of phage-resistant strain R3 was slightly slower than that of host strain; biochemical and virulent genetic tests showed that there was no difference between the resistant strain and host strain; compared to the host strain, the pathogenicity of the resistant strain to mice (Lethal Dose, 50%, LD₅₀) increased by 74.8% (P>0.05); and the adhesion ability to MODE-K cells were slightly weaker, but the difference was not significant. [Conclusion] Compared with the host strain, the biological characteristics and virulence genes of the bacteriophage-resistant strains did not change, and the pathogenicity of the strains was weakened in mice, but the adhesion ability to MODE-K cells was not significantly different.

Abstract:[Objective] To explore the effect of Clostridium butyricum on pathogenicity and protection mechanism of Salmonella enteritidis (CVCC3377) in SPF mice. [Methods] Mice were sacrificed on the 6th day post infection and samples of liver, spleen and cecum samples were collected. The pathological changes of liver and spleen tissue were detected by H.E staining, the expression levels of TLR4 and MyD88 protein in spleen tissue were detected by immunohistochemistry, and the expression levels of TNF-α and IL-6 in cecum tissue were detected by fluorescence quantitative PCR. The effect on intestinal flora was analyzed by high-throughput sequencing. [Results] C. butyricum could alleviate the weight loss of mice caused by Salmonella enteritidis infection. The results of HE staining show that Clostridium butyricum could alleviate the tissue damaged by Salmonella enteritidis infection. The results of immunohistochemistry show that Clostridium butyricum could decrease the expression levels of TLR4 and MyD88 protein in liver and spleen tissue cells. Fluorescence quantitative PCR shows that Salmonella enteritidis could significantly increase TNF-α in cecal tissue (P<0.05). The expression of TNF-alpha and IL-6 decreased by feeding Clostridium butyricum. The results of high throughput sequencing show that the Chao 1 index, ACE index and Shannon index of Clostridium butyricum treatment group were higher than those of the control group and Salmonella infection group (P>0.05). Clostridium butyricum could significantly reduce the proportion of Actinobacteria and Verrucomicrobia in intestinal microorganisms (P<0.05). In addition, the relative abundance of Lactobacillus in intestinal microbial community increased by bacteria (P>0.05). [Conclusion] Clostridium butyricum could improve the immunity of mice, alleviate the inflammatory reaction caused by Salmonella infection, increase intestinal microflora diversity and regulate intestinal flora structure.

Abstract:[Objective] A lead and cadmium resistant strain was isolated from the polluted soil of Zhalong wetland, and the influences on lead and cadmium adsorption under different conditions are discussed. [Methods] The streak plate method and progressive domestication were used for screening lead and cadmium resistant strain. The strain was identified by physiological and biochemical tests, and by ITS sequence analysis. The optimal condition of the lead and cadmium resistant strain was explored, and Langmuir and Freundlich isothermal adsorption models were used. [Results] A lead and cadmium resistant strain JB15 was isolated as Beauveria bassiana, and the maximum tolerant Pb²⁺ and Cd²⁺ concentration of the strain was 1200 mg/L and 200 mg/L, respectively. Optimum conditions for adsorption were found at pH of 7.0 with inoculation quantity of 8.0 g/L, at 30℃ for 60 min. Under such conditions, the lead and cadmium adsorption rate reached 52.27% and 62.38%, and the lead and cadmium adsorption capacity reached 19.60 mg/g and 3.98 mg/g, respectively. The adsorption process of the strain was fitted well on Langmuir adsorption isotherm model. [Conclusion] The isolated strain JB15 has a good adsorption effect and provides theoretical foundation for microbial remediation of soil heavy metal contamination.

Abstract:[Objective] We characterized the function of cntRLMN in zinc ion uptake of Pseudomonas aeruginosa PAO1 (ATCC15692). [Methods] On the basis of the mutant strain ΔznuBC, we built ΔcntRLMN using homologous recombination. We also constructed its complementary strain and lacZ fusion report strain through plasmid conjugation transfer. Transcriptional regulation of Zur protein on cntRLMN was studied by beta-galactoside enzyme activity assay. In vitro binding of Zur protein to cnt promoter and mutated fragments of cnt promoter was tested using EMSA. The zinc ion uptake function of cntR, cntL, cntN and other genes in the cntRLMN operon was examined through growth curve analysis. Finally, the influence of cntRLMN mutation on the virulence of P. aeruginosa was studied using the infection model of the wax moth larva. [Results] The enzyme activity analysis of lacZ transcription and fusion showed that cntRLMN was negatively regulated by Zur protein, and its expression was induced by zinc ion starvation in a Zur-dependent manner. The EMSA results showed that the promoter of cntRLMN could bind His-Zur to form a DNA-protein complex, where the binding site was GCGTTATAGTATATCAT. The growth curve and the infection experiment of the wax moth larva showed that ZnuBC and CntRLMN were functionally complementary; when grown in Zn-restricted culture, the toxicity of P. aeruginosa to the wax moth larve was significantly inhibited only in the znuBC and cntRLMN double-deleted strain. [Conclusion] The cntRLMN, which plays an important role in toxicity of P. aeruginosa, is likely an alternative zinc ion uptake system directly and negatively regulated by the Zur protein.

Abstract:[Objective] The Th1 and Th2-type immune responses induced by the Brucella gntR gene were analyzed by prokaryotic expression of gntR. [Methods] To clone and express gntR gene of Brucella, a pair of primers for the gntR gene of Brucella S2308 was designed based on published GenBank sequence. The gntR gene was amplified by molecular cloning technology from Brucella S2308 and then cloned into pET-30a vector and transformed to E. coli BL21. The expression of GntR protein was analyzed by SDS-PAGE. The GntR recombinant protein (rGntR) was purified by AKTAxpress intelligent multidimensional purification system. The rGntR immunoreactivity was analyzed by Western blotting. Cytokines IFN-γ, IL-2, IL-4 and IL-5 in murine macrophages (RAW 264.7) and splenocytes stimulated with rGntR were detected by ELISA kit. Mice were immunized with rGntR and IFN-γ, IL-4 and IgG in serum of mice were detected by ELISA kit. [Results] The full-length gntR gene was 735 bp, encoding 245 amino acids. SDS-PAGE showed that rGntR appeared in the approximately 35 kDa position. The band was single after purification. WB indicated that rGntR had good immunoreactivity. The rGntR could induce higher levels of IFN-γ, IL-4 and IgG in host cells and mice. [Conclusion] The rGntR could induce helper T cells (Th1 and Th2-type) immune responses in vivo and in vitro. This study provides scientific basis for the development of brucellosis vaccines.

Abstract:[Objective] To obtain complete sequence of an integrating conjugative element (ICE), ICEVpaTF2, from Vibrio parahaemolyticus TF2, and analyze its genomic characteristics; to investigate if ICEVpaTF2 has excision/cyclization activities from host genome and how att sites recombine during excision/cyclization. [Methods] The genomic frame sequences of V. parahaemolyticus TF2 was first annotated by RAST, which led to the finding of a potential and complete ICE element, named ICEVpaTF2. After manual splicing, PCR amplification followed by sequencing verification, a complete sequence of ICEVpaTF2 was obtained, and then it was annotated and characterized again. PCR detection and subsequent sequencing were performed to explore whether ICEVpaTF2 could excise from its host genome and cyclize itself and to characterize the forming of attB and attP by comparing the sequences of attL, attR, attB and attP. [Results] ICEVpaTF2, with a total length of 83588 bp, contains 52 conserved core genes of SXT/R391 ICE elements, which are related to excision, integration, self-transfer, and regulatory mechanisms. ICEVpaTF2 also contains five hot spots (HS), two variable regions (VR), and three atypical insertion sites accommodating variable genes. HS and VR contain a large number of variable genes, encoding for restrictive modification system, DNA repair system, toxin-antitoxin system, and etc., which confer the host extensive adaptive profits to the bacterial host. ICEVpaTF2 also contains unique genes with unknown functions. Through phylogenetic analysis of the two core conserved genes, int and xis, it was found that int and xis of ICEVpaTF2 belong to subgroups represented by that of R391 and SXT, respectively. ICEVpaTF2 excises at chromosomal attL and attR sites and cyclizes resulting in the forming of new heterozygous and recombined attB and attP sites. [Conclusion] ICEVpaTF2 belongs to the SXT/R391 family and it is a complete ICE element harboring self-excision and cyclization activities. New attB and attP sites are generated by hybrid recombination between primary attL and attR sites.

Abstract:[Objective] Intestinal bacteria play an important role in host metabolism and health. It has been reported that the intestinal flora of Gansu zokor is related to regional, gender and seasonal factors, however, it is not clear whether there is a difference in the intestinal bacterial of Gansu zokor feeding with different feeds under wild and artificial feeding conditions. [Methods] In this study, we selected alfalfa, the roots of Mongolian, Chinese pine and spruce for feeding groups, wild Gansu zokors captured in the same habitat as controls. Bacterial diversity of digestive tract contents in Gansu zokor was analyzed by 16S rRNA V3-V4 high-throughput sequencing technology. [Results] Under the conditions of wild and artificial feeding, the dominant bacteria were both Firmicutes and Bacteroidetes, but the content was significantly different. Under artificial feeding conditions, the intestinal bacterial community structure of Mongolian pine, Chinese pine and spruce was similar, they all had significant difference with alfalfa and wild groups. The intestinal bacterial diversity of Mongolian pine, Chinese pine, spruce were higher than alfalfa and wild groups. [Conclusion] The results showed that there were significant differences in the composition and structure of intestinal bacterial of Gansu zokor under wild and artificial feeding conditions.