The present study aimed to isolate an optimal lactic acid bacterial strain from the feces of healthy giant pandas. The strain exhibited good stability at low pH and high bile salt concentrations, activity against pathogens relevant to pandas, and antibiotic susceptibility. In the current study, 25 isolates were obtained from de Man, Rogosa, and Sharpe agar. Two (E21 and G83) and eight (E1, E2, E16, E18, E21, E69, E70, and G83) isolates demonstrated good performance at pH 2.0 and bile 2% (w/v), respectively. Three isolates (G83, G88, and G90) possessed better antimicrobial effect on enterotoxigenic Escherichia coli CVCC196 (ETEC) than the rest. One isolate (G83) strongly affected Salmonella, whereas three (G83, G87, and G88) exhibited inhibitory activity against Staphylococcus aureus. All isolates were multi-drug resistant. These isolates were identified as Lactobacillus (5 isolates) and Enterococcus (20 isolates) by 16S rRNA sequencing. Virulence genes were detected in Enterococcus isolates. Isolate G83 was identified as Lactobacillus plantarum and was considered as the best probiotic candidate among all of the experimental isolates. This study provided necessary and important theoretical guidance for further experiments on G83 in vivo.

In this work, we searched for an effective probiotic that can help control intestinal infection, particularly enterotoxigenic Escherichia coli K88 (ETEC) invasion, in giant panda (Ailuropoda melanoleuca). As a potential probiotic strain, Lactobacillus plantarum BSGP201683 (L. plantarum G83) was isolated from the feces of giant panda and proven beneficial in vitro. This study was aimed to evaluate the protective effect of L. plantarum G83 in mice challenged with ETEC. The mice were orally administered with 0.2 mL of PBS containing L. plantarum G83 at 0 colony-forming units (cfu) mL-1 (control; negative control, ETEC group), 5.0 × 108 cfu mL-1 (LDLP), 5.0 × 109 cfu mL-1 (MDLP), and 5.0 × 1010 cfu mL-1 (HDLP) for 14 consecutive days. At day 15, the mice (LDLP, MDLP, HDLP, and ETEC groups) were challenged with ETEC and assessed at 0, 24, and 144 h. Animal health status; chemical and biological intestinal barriers; and body weight were measured. Results showed that L. plantarum G83 supplementation protected the mouse gut mainly by attenuating inflammation and improving the gut microflora. Most indices significantly changed at 24 h after challenge compared to those at 0 and 144 h. All treatment groups showed inhibited plasma diamine oxidase activity and D-lactate concentration. Tight-junction protein expression was down-regulated, and interleukin (IL)-1β, IL-6, IL-8, TLR4, and MyD88 levels were up-regulated in the jejunum in the LDLP and MDLP groups. The number of the Enterobacteriaceae family and the heat-labile enterotoxin (LT) gene decreased (P < 0.05) in the colons in the LDLP and MDLP groups. All data indicated that L. plantarum G83 could attenuate acute intestinal inflammation caused by ETEC infection, and the low and intermediate doses were superior to the high dose. These findings suggested that L. plantarum G83 may serve as a protective probiotic for intestinal disease and merits further investigation.

为了解成都地区犬布氏杆菌病的流行情况并建立一种快速、可靠、特异、灵敏的布氏杆菌的检测方法,对来自宠物医院、狗场等的860份犬血清样品进行了血清学检测,并针对布氏杆菌属保守基因设计1对特异性引物并建立了一种基于SYBR Green的荧光定量PCR检测方法.血清学检测结果显示,成都地区存在犬布氏杆菌病的总阳性率为1.51％,其中流浪狗阳性率最高,其次是狗场,宠物医院犬布氏杆菌病的阳性率最低.本研究建立的荧光定量PCR具有较高的特异性,标准曲线相关系数为0.996,扩增效率为91％,能够用于未知样品的定量检测.用建立的荧光定量PCR检测了33份虎红平板凝集试验(RBPT)阳性血清,结果,其中10份呈阳性;检测了30份随机抽取的阴性血清,其中2份呈阳性.结论:本研究建立的荧光定量PCR与RBPT的检测结果有差异,两种方法均检测到成都地区犬布氏杆菌病.

Accumulating evidence has proven the immunomodulating activity of Yupingfeng. This study compared the immunomodulatory activity in vitro of the unfermented Yupingfeng dreg polysaccharides (UYDP) with that of the fermented Yupingfeng dreg polysaccharides (FYDP) obtained using Rhizopus oligosporus SH. Results consistently elucidated the duality of the immunomodulatory roles of UYDP and FYDP in regulating proliferation, and cytokines expressions in murine lymphocytes and macrophages. Compared with UYDP, FYDP effectively enhanced the proliferation of lymphocytes and promoted mRNA expression of inducible nitric oxide synthase (iNOS), IL-6, TNF-α, INF-γ, nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB), and TLR-4 in lymphocytes and macrophages. Moreover, compared with UYDP, FYDP effectively normalized cell proliferation and downregulated mRNA expression levels of pro-inflammatory cytokines, NF-κB, TLR-4, and iNOs in lipopolysaccharide-induced chronic inflammation cells. The results revealed that the bidirectional immunomodulatory effects in vitro of UYDP and FYDP, and the bi-directional immunomodulatory activity of FYDP is superior over that of UYDP. Moreover, more studies in vivo that needs to be studied further.

Yupingfeng (YPF) is a kind of Astragali radix-based ancient Chinese herbal supplemented with Atractylodis Macrocephalae Rhizoma and Radix Saposhnikoviae. Increasing evidence has proven the beneficial immunomodulating activity of YPF. However, the action mechanism(s) of it is not known. Here, we explored the immunomodulatory activity of unfermented Yupingfeng polysaccharides (UYP) and fermented Yupingfeng polysaccharides (FYP) obtained using Rhizopus oligosporus SH in weaning Rex rabbits. The results showed that both UYP and FYP exhibited notable growth-promoting and immune-enhancing activities, improvement of the intestinal flora homeostasis, and maintenance of intestinal barrier integrity and functionality. Notably, compared with UYP, FYP effectively enhanced average daily gain, organ indices, interleukin-2 (IL-2), IL-4, IL-10, tumor necrosis factor-alpha (TNF-α), TLR2, and TLR4 mRNA levels in spleen, IL-1, IL-2, IL-4, IL-6, IL-10, IL-12, TNF-α, and IFN-γ protein concentrations in serum, and TLR2 and TLR4 mRNA expressions in the gastrointestinal tract (GIT). Moreover, FYP exhibited greater beneficial effects in improving the intestinal flora, including augment flora diversity and the abundance of cellulolytic bacteria, reduction the abundance of Streptococcus spp. and Enterococcus spp. in the GIT, particularly the foregut and maintaining the intestinal barrier integrity and functionality by upregulating zonula occludens 1, claudin, polymeric immunoglobulin receptor, trefoil factor, and epidermal growth factor mRNA levels in the jejunum and ileum. Our results indicated the immunoenhancement effect of FYP is superior over that of UYP, which is probably related with the amelioration of the intestinal microflora and intestinal barrier in the foregut.

Antibiotics have been widely used for the prevention and the treatment of diseases to humans and animals, and they have fed additives for agricultural animals to promote growth. However, there is a growing concern over the practice due to its side effects on intestinal microbial communities which plays a vital role in animals' health. To investigate the effect of antibiotics on the bacterial population of the caecum in rex rabbits, 80 rex rabbits were randomly divided into four groups: control group (B, basal diet), chlortetracycline group (C, 50 mg/kg), colistin sulfate group (S, 20 mg/kg) and zinc bacitracin group (Z, 40 mg/kg). Caecum microbial communities of rex rabbits from the four groups were analyzed through Illumina Miseq platform after being fed 28 days. The results showed that most obtained sequences belongs to Firmicutes followed by Bacteroidetes, and the ratio of Bacteroidetes/Firmicutes in C group (42.31 %) was higher than that in Z group (21.84 %). Zinc bacitracin supplementation caused a significant decreased of the Proteobacteria phylum and Lactobacillus spp. (P < 0.05), while the Lactobacillus spp. significantly increased in S group (P < 0.05). In addition, Ruminococcus spp., especially Ruminococcus albus were the predominant bacterial species found in both S and Z groups. The proportion of Coprococcus spp. significantly increased in Z group (P < 0.05). These findings suggested that the antibiotics used may cause significant changes in the caecum microbiota of rex rabbits, and we also found C group had a similarity caecum bacteria structure with B group which was probably due to the high levels of chlortetracycline resistance.

It has been established beyond doubt that giant panda genome lacks lignin-degrading related enzyme, gastrointestinal microbes may play a vital role in digestion of highly fibrous bamboo diet. However, there is not much information available about the intestinal bacteria composition in captive giant pandas with different ages. In this study, we compared the intestinal bacterial community of 12 captive giant pandas from three different age groups (subadults, adults, and geriatrics) through PCR-denaturing gradient gel electrophoresis (DGGE) and real-time PCR analysis. Results indicated that microbial diversity in the intestine of adults was significantly higher than that of the geriatrics (p<0.05), but not significant compared to the subadults (p>0.05). The predominant bands in DGGE patterns shared by the twelve pandas were related to Firmicutes and Proteobacteria. Additionally, in comparison to healthy individuals, antibiotic-treated animals showed partial microbial dysbiosis. Real-time PCR analyses confirmed a significantly higher abundance of the Lactobacillus in the fecal microbiota of adults (p<0.05), while other bacterial groups and species detected did not significantly differ among the three age groups (p>0.05). This study revealed that captive giant pandas with different ages showed different intestinal bacteria composition.

This test was conducted to study the effects of the Lactobacilli and bursin on growth performance, slaughter performance, meat quality and serum biochemical parameters of broilers by adding Lactobacilli prepa-ration in a basal diet or intramuscular injection bursin.One hundred and eighty 1-day-old Cobb broilers were randomly divided into 4 groups with 3 replicates in each group and 15 broilers per replicate.The broilers in group I were fed the basal diet, those in group Ⅱ were fed the basal diet and injected bursin ( 0.01 mg/kg BW) , those in group Ⅲwere fed the basal diet+0.1%Lactobacilli preparation, and those in group Ⅳ were fed the basal diet+0.1%Lactobacilli preparation, and injected bursin ( 0.01 mg/kg BW) .The test lasted for 42 d.The results showed as follows: 1 ) the average daily feed intake in groups Ⅱ and Ⅳ was significantly higher than that in groupsⅠ and Ⅲ ( P<0.01) , but the average daily gain and feed/gain were no significant difference among groups (P>0.05).2) The semi-eviscerated rate in group Ⅱ was significantly higher than that in groupⅠ (P<0.01), and the slaughter rate, eviscerated rate, breast muscle rate and leg muscle rate were no significant difference among groups ( P>0.05) .3) The meat color in groupsⅡandⅢwas much bet-ter than that in groupsⅠandⅣ, but the pH was no significant difference among groups ( P>0.05) .The drip loss in groupsⅡandⅣwas significantly higher than that in groupsⅠandⅢ( P<0.01) , and the shear force in groups Ⅱ,Ⅲ and Ⅳwas significantly lower than that in groupⅠ (P<0.01).4) The surem total protein and albumin contents in groupⅢwere significantly higher than those in groupⅠ( P<0.05) and the surem tri-glyceride content in groups Ⅲ and Ⅳ was significantly lower than that in groupⅠ( P<0.05) .The composite indicators show that Lactobacilli and bursin can improve the growth performance and slaughter performance of broilers to a certain extent, and can significantly improve meat quality and serum biochemical parameters of broilers; furthermore, the combination effect of Lactobacilli and bursin is not superior to single use.

Purpose: The objective of this study is to evaluate the regulatory effect of Lactobacillus plantarum F1208 on gastrointestinal functions in constipated rats.Methods: Male Wistar rats were randomly divided into health group,constipation group,senna group,and Lactobacillus plantarum high-,middle-and low-dosage groups.A rat model of loperamide hydrochloride-induced constipation was established.Each group was treated with corresponding drugs at 2 mL/(kg.d) for 7 days.During the experiment,feed intake,water intake,body quality increase,fecal number and water content,live bacterial count in feces and gastrointestinal transit ratio were monitored.Results: The administration with Lactobacillus plantarum F1208 for 7 days in constipated rats significantly improved the feed intake and water intake,promoted the growth and development of rats,increased the number and water content of feces within 24 hours,restrained the growth of intestinal bowel enterobacter and enterbacteriaceae,and improved gastrointestinal transit ratio.Conclusion: Lactobacillus plantarum F1208 has improved gastrointestinal functions in constipated rats.Conclusion: Lactobacillus plantarum F1208 has improvement gastrointestinal function in constipation rats.

To investigate effects of Bacillus licheniformis on ileal tissue and cytokines in chicken with necrotic enteritis,one hundred twenty 1day-old male chickens were randomly divided into control group,infection group,prevention group(1g/kg BacillusⅠ)and treatment group(0.5g/kg BacillusⅠ+4g/kg BacillusⅡ).The infection group,the prevention group and the treatment group were given with coccidian on 15day-old,followed by C.perfringens(1×108 CFU/mL)from 17days-old to 20days-old.The expression levels of IFN-α,IFN-γ,IL-1βand IL-6in ileum were detected by the real-time PCR,and the intestinal histological changes were observed by tissue section.In result,the infected group showed markedly increased peripheral blood leukocytes and ileum cytokine expression level.There was obvious lesions in the ileum tissue.The supplementation of Bacillus licheniformis reduced the intestinal lesions and peripheral blood leukocytes.Moreover,compared with the infection group,the expression levels of IFN-γ,IL-1βand IL-6were significantly reduced in the treatment group,while only IL-1βreduced in the prevention group,and no significant difference in the other cytokines.The result showed that Bacilluslicheniformishave positive effect on the prevention of necrotic enteritis in chicken.

To explore the defecating function of fructooligosaccharide(FOS) and Lactobacillus,constipation model of Wistar rats was established.From 36 male wistar rats of the same age and weight,4 rats were randomly selected as blank control group,while the remaining rats were used as constipation model and randomly divided into 8 groups,including model group,positive treatment group(folium senna 10.0 mg/kg),FOS high,medium and low-dose groups(corresponding to 1.0,0.5,0.25 g/(kg.d) and Lactobacillus high,medium and low-dose groups(corresponding to 2.0 × 1010,2.0 × 109,2.0 × 108 CFU/(kg.d)).After continuous force-feeding for 7 days,6 indexes,i.e.,feed intake,water intake,body mass increment,the advance rate of carbon powder,fecal pellets and intestinal flora,were detected.Both FOS and Lactobacillus had defecating function.Water intake,body mass increment,the advance rate of carbon powder and fecal pellets of the 6 groups with gavage of FOS or Lactobacillus were significantly higher than those of model group(P 0.05).Besides,FOS and Lactobacillus had an inhibiting effect on the growth of intestinal Enterococci and Enterobacteriaceae(P 0.05).The optimal dosage of FOS and Lactobacillus was 1.0 g/(kg.d) and 2.0 × 108 CFU/(kg.d),respectively.Both FOS and Lactobacillus have defecating function,maintain the balance of intestinal flora and inhibit the growth of intestinal Enterococci and Enterobacteriaceae.