Aims: This study aimed to evaluate antidiabetic potential of indigenous Lactobacillus isolates by measuring the ability of α-glucosidase inhibitory (AGI) and antioxidant activity. The mechanism of probiotics as antidiabetic can occur through the AGI and antioxidant activity of LAB, which is able to suppress oxidative stress that causes chronic inflammation and pancreatic β cell apoptosis, and then through the ability to produce exopolysaccharide (EPS) and short chain fatty acids (SCFA). Methodology and results: MRS broth enriched with 10% glucose was selected as the growth medium for Lactobacillus. The growth medium was then centrifuged to obtain CFS and CFE was produced by extracting the medium with 96% ethanol as a solvent. The results showed that Lactobacillus pentosus MK42 had the highest AGI activity of 80.32 ± 2.20%. Antioxidant activity was not significantly different (P>0.05) among the tested Lactobacillus isolates. Lactobacillus paracasei RK41 produced the highest EPS (360.13 ± 50.01 mg/L), which was not significantly different (P>0.05) from Lactobacillus plantarum1 RB210. All Lactobacillus isolates were able to produce acetic acid, but not all were able to produce propionic and butyric acid. The highest propionic acid was produced by L. plantarum1 RB210 at 0.40 ± 0.31 mmol/L and the highest butyric acid was produced by L. plantarum1 MK2 at 0.22 ± 0.08 mmol/L. Conclusion, significance and impact of study The results show definitively that indigenous Lactobacillus isolates have considerable α-glucosidase inhibitor, antioxidant activity and the ability to produce of EPS and SCFA. This preliminary study suggests the use of indigenous Lactobacillus isolates which have the potential as antidiabetic agent, although the responsible compounds are unknown.
alpha-Glucosidases ; Antioxidants ; Lactobacillus--isolation & ; purification ; Hypoglycemic Agents

OBJECTIVETo investigate the vaginal flora in patients with recurrent vulvovaginal candidiasis (RVVC).

METHODSVaginal swabs were collected at different time points from 6 RVVC patients and 5 healthy women of child-bearing age. The dynamic changes, microbiota composition, alpha diversity and beta diversity in the two groups were assessed by analyzing the 16S rRNA V4 hypervariable region amplified from the total genomic DNA from the swabs.

RESULTSLactobacillus was the predominant species in healthy women with similar proportions of L.iners and L.crispatus; small proportions of Gardnerella, Prevotella and other genus were also detected. In some healthy women, the vaginal flora showed a high relative abundance of anaerobic bacteria such as Gardnerella, Prevotella, Atopobium, Sneathia. Compared with the healthy women, patients with RVVC showed a significantly reduced diversity of vaginal flora, where L.iners was the predominant species and the content of L.crispatus decreased significantly. In healthy women, the vaginal flora fluctuated with the menstrual cycle, and the fluctuation was the most prominent during menstruation; the dominant species either alternated regularly or maintain an absolute superiority in the menstrual cycle. The vaginal flora showed attenuated fluctuation in women with RVVC, were highly conserved within the menstrual cycle, and maintained a similar composition in the episodes and intermittent periods.

CONCLUSIONThe vaginal flora of RVVC patients do not undergo regular variations with the menstrual cycle and shows a similar composition between the episodes and intermittent periods. Promoting the production of L.iners or inhibiting the colonization of L.crispatus to restore the composition of the vaginal flora may help in the treatment of RVVC.

Candidiasis, Vulvovaginal ; microbiology ; Case-Control Studies ; Female ; Humans ; Lactobacillus ; classification ; isolation & purification ; Longitudinal Studies ; Menstrual Cycle ; Microbiota ; RNA, Ribosomal, 16S ; isolation & purification ; Vagina ; microbiology

OBJECTIVETo analyze the whole microbial structure in a case of rampant caries to provide evidence for its prevention and treatment.

METHODSClinical samples including blood, supragingival plaque, plaque in the caries cavity, saliva, and mucosal swabs were collected with the patient's consent. The blood sample was sent for routine immune test, and the others samples were stained using Gram method and cultured for identifying colonies and 16S rRNA sequencing. DNA was extracted from the samples and tested for the main cariogenic bacterium (Streptococcus mutans) with qPCR, and the whole microbial structure was analyzed using DGGE.

RESULTSThe patient had a high levels of IgE and segmented neutrophils in his blood. Streptococci with extremely long chains were found in the saliva samples under microscope. Culture of the samples revealed the highest bacterial concentration in the saliva. The relative content of hemolytic bacterium was detected in the samples, the highest in the caries cavity; C. albicans was the highest in the dental plaque. In addition, 33 bacterial colonies were identified by VITEK system and 16S rDNA sequence phylogenetic analysis, and among them streptococci and Leptotrichia wade were enriched in the dental plaque sample, Streptococcus mutans, Fusobacterium nucleatum, and Streptococcus tigurinus in the caries cavity, and Lactobacillus in the saliva. S. mutans was significantly abundant in the mucosal swabs, saliva and plaque samples of the caries cavity as shown by qPCR. Compared to samples collected from a healthy individual and another two patients with rampant caries, the samples from this case showed a decreased bacterial diversity and increased bacterial abundance shown by PCR-DGGE profiling, and multiple Leptotrichia sp. were detected by gel sequencing.

CONCLUSIONThe outgrowth of such pathogenic microorganisms as S. mutans and Leptotrichia sp., and dysbiosis of oral microbial community might contribute to the pathogenesis of rampant caries in this case.

Abnormalities, Multiple ; Dental Caries ; microbiology ; Dental Plaque ; microbiology ; Fusobacterium ; isolation & purification ; Humans ; Immunoglobulin E ; blood ; Lactobacillus ; isolation & purification ; Leptotrichia ; isolation & purification ; Limb Deformities, Congenital ; Microbiota ; Mouth Mucosa ; microbiology ; Neutrophils ; cytology ; Phylogeny ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S ; genetics ; Saliva ; microbiology ; Streptococcus ; isolation & purification ; Tooth Abnormalities

OBJECTIVETo evaluate the effect of water pollution with dexamethasone on intestinal flora in mice.

METHODSTwenty Balb/c mice were randomly divided into control group and low-, moderate- and high-dose dexamethasone groups. The mice in dexamethasone groups were exposed to dexamethasone sodium phosphate in drinking water at doses of 0.035, 0.225, and 2.25 ng for 36 days. The changes in behaviors, fur condition, and feces of the mice were observed daily. All the mice were sacrificed at 36 days and the tissues in the ileocecal region was collected for denaturant gradient gel electrophoresis (DGGE) of 16S rDNA V6 variable regions of microbes and sequence analysis with BLAST.

RESULTSThe mice in the 3 dexamethasone groups all showed aggressive behaviors. Cluster analysis of DGGE graph showed relatively stable floras in the ileocecal region in all the mice, but principal component analysis identified differences in the dominating flora among the groups. Diversity analysis of the flora revealed significantly increased amount and types of bacteria in the intestinal flora in all the 3 dexamethasone groups (P<0.05 or 0.01) compared with the control group. Sequence analysis of 16S rDNA V6 regions showed 15 common bacterial species and 2 differential species between the dexamethasone groups and the control group with changes in the type and proportion of the dominating bacterium in the dexamethasone groups. Lactobacillus colonization was detected in the control group but not in moderate- and high-dose dexamethasone groups, and Shigella species were found in the latter two groups.

CONCLUSIONSWater contamination with dexamethasone can affect the nervous system of mice, cause changes in the types and amounts of intestinal bacteria and the dominating bacteria, and inhibit the colonization of probiotics in the intestinal floras to increase the risk of invasion by intestinal pathogenic bacteria.

Animals ; Bacteria ; classification ; Dexamethasone ; pharmacology ; Drinking Water ; chemistry ; Feces ; Gastrointestinal Microbiome ; drug effects ; Lactobacillus ; isolation & purification ; Mice ; Mice, Inbred BALB C ; Probiotics ; RNA, Bacterial ; genetics ; RNA, Ribosomal, 16S ; genetics ; Shigella ; isolation & purification

OBJECTIVE: To investigate the risky factors for pelvic floor functional disorder.
 METHODS: A total of 346 patients, who undergone surgery for pelvic floor reconstruction from January 2007 to December 2013 in the Third Affiliated Hospital of Zhengzhou University, were enrolled for this study and served as an experimental group. Meanwhile, 349 female healthy subjects were selected as a control group. The data including the times of gestation and parity, history of menopause, hysterectomy surgery, uterine fibroid and leucorrhea were recorded.
 RESULTS: Compared with the controls, the numbers of people with hysterectomy, menopause and clean vagina≥III degree, lack of lactobacillus as well as the times of gestation and parity in the experimental group were significantly increased (P<0.05). The history of hysterectomy, the times of gestation and parity and lack of lactobacillus in vagina were the independent risk factors of pelvic floor dysfunction.
 CONCLUSION The history of hysterectomy, the times of gestation and parity, and clean vagina may involve in the occurrence and development of pelvic floor functional disorder, and the existence of lactobacillus in the vagina might be a protective factor.
Case-Control Studies ; Female ; Humans ; Hysterectomy ; Lactobacillus ; isolation & purification ; Menopause ; Multivariate Analysis ; Parity ; Pelvic Floor ; physiopathology ; Pelvic Floor Disorders ; physiopathology ; Pregnancy ; Risk Factors ; Vagina ; microbiology

Bacteria ; genetics ; isolation & purification ; Bacterial Physiological Phenomena ; Bacteroidetes ; genetics ; isolation & purification ; Body Mass Index ; Body Weight ; Child ; Energy Metabolism ; Humans ; Infant ; Intestines ; metabolism ; microbiology ; Lactobacillus ; Obesity ; etiology ; metabolism ; prevention & control ; Probiotics ; therapeutic use ; Risk Factors

China ; Drug Resistance, Bacterial ; Electrophoresis, Gel, Pulsed-Field ; Fast Foods ; microbiology ; Lactobacillus ; isolation & purification ; physiology ; Microbial Sensitivity Tests ; RNA, Ribosomal, 16S ; genetics ; Streptococcus ; isolation & purification ; physiology ; Yogurt ; microbiology

OBJECTIVETo investigate the changes in fecal flora and its correlation with the occurrence and progression of inflammatory bowel disease (IBD).

METHODSWe collected fresh fecal specimens from 167 IBD patients (including 113 with ulcerative colitis and 54 with Crohn's disease) and 54 healthy volunteers. The fecal flora was analyzed by gradient dilution method and the data of inflammatory markers including WBC, PLT, CRP and ESR were collected to assess the association between the fecal flora and the inflammatory markers.

RESULTSThe species Enterrococcus (6.60∓0.23, P<0.01), Saccharomyces (2.22∓0.27, P<0.05), Bacteriodes (5.57∓0.28, P<0.001), Bifidobacterium (5.08∓0.30, P<0.01), Peptococcus (6.22∓0.25, P<0.001), Lactobacillus (6.00∓0.26, P<0.001), and Clostridium (3.57∓0.30, P<0.05) all increased significantly, while Eubacterium (1.56∓0.24, P<0.01) reduced markedly in patients with ulcerative colitis compared with those in the control subjects. Enterrococcus (6.93∓0.28, P<0.01), Saccharomyces (2.73∓0.37, P<0.01), Bacteriodes (4.32∓0.52, P<0.05), Bifidobacterium (4.88∓0.42, P<0.05), Peptococcus (6.19∓0.32, P<0.01) and Lactobacillus (4.73∓0.47, P<0.001) all increased significantly and Eubacterium (1.01∓0.29, P<0.01) and Clostridium (0.87∓0.31, P<0.01) decreased in patients with Crohn's disease. The positivity rates of bacterial culture were consistent with the results of quantitative analysis of the fecal flora. The changes in fecal flora did not show a significant correlation with these inflammatory markers.

CONCLUSIONIBD patients have fecal flora imbalance compared with the healthy controls, and this imbalance may contribute to the occurrence and progression of IBD. The decline of Eubacterium contributes to the occurrence and development of IBD.

Adult ; Bacteria ; isolation & purification ; Bacteroides ; isolation & purification ; Bifidobacterium ; isolation & purification ; Biomarkers ; analysis ; Clostridium ; isolation & purification ; Colitis, Ulcerative ; microbiology ; Crohn Disease ; microbiology ; Enterococcus ; isolation & purification ; Eubacterium ; isolation & purification ; Feces ; microbiology ; Female ; Humans ; Inflammatory Bowel Diseases ; etiology ; microbiology ; Lactobacillus ; isolation & purification ; Male ; Peptococcus ; isolation & purification ; Saccharomyces ; isolation & purification

We selected and characterized isolates of Lactobacillus crispatus (L. crispatus) for potential preventing infections of the female reproductive tract. We cultured vaginal swabs from healthy volunteers on de Man, Rogosa and Sharpe (MRS) agar and identified the isolates at the species level by 16S rRNA sequence and genotyped the isolates of Lactobacillus by PCR amplification of repetitive bacterial DNA elements (rep-PCR). Furthermore, 10 L. crispatus strains were assessed for hydrogen peroxide (H2O2) and acid production. Overall 65 isolates were confirmed to be Lactobacillus by sequence analogy, among them 19 were L. crispatus, 17 were Lactobacillus jensenii and 12 were Lactobacillus fermentum. rep-PCR produced specie and strain-specific genomic fingerprints for the Lactobacillus isolates. The selected 10 L. crispatus isolates produced highly acidic environment after growth in MRS. The isolates T22-3 and T29-5 demonstrated high production of H2O2. This study indicated that there are individual differences with vaginal Lactobacillus colonization, and strain diversity within vaginal L. crispatus isolates, T22-3 and T29-5 might be candidates for restoring urogenital health environment in females.
Adult ; Female ; Genotype ; Humans ; Hydrogen Peroxide ; metabolism ; Interspersed Repetitive Sequences ; Lactobacillus ; classification ; genetics ; isolation & purification ; physiology ; Polymerase Chain Reaction ; methods ; Vagina ; microbiology

OBJECTIVETo develop a pulsed field gel electrophoresis (PFGE) method for molecular typing of Lactobacillus and Streptococcus thermophilus (S. thermophilus) and to apply it in identification and characterization of both bacteria isolated from yoghurt collected from Beijing supermarket.

METHODSThe five most useful restriction enzymes including Apa I, Not I, Sfi I, Xba I and Sma I were chosen to cut DNA of 52 strains of Lactobacillus, S. thermophilus as well as associated standard bacteria strains. The endonucleases and electrophoresis conditions for PFGE analysis were optimized and applied in molecular typing of Lactobacillus and S.thermophilus isolates. Cluster analysis based on the PFGE data was conducted. The identification results of PFGE were compared with those obtained in biochemical and 16s ribosomal RNA PCR identification tests.

RESULTSNot I was suitable for L. bulgaricus, L. fermentum and L. delbrueckii digestion. While Apa I was an appropriate endonuclease for S. thermophilus, L. acidophilus and L. casei digestion. The results of molecular typing indicated that 24 strains of L.bulgaricus and 15 strains of S. thermophilus were grouped into 8 types by PFGE method, respectively. While 7 strains of L.acidophilus were grouped into 3 types and 2 strains of L. delbrueckii were grouped into 2 different PFGE types.

CONCLUSIONThe results of PFGE analysis are in compliance with those of 16s rRNA PCR and biochemical identification. The PFGE method developed in this study is suitable for molecular characterization of both Lactobacillus and S. thermophilus.

Bacterial Typing Techniques ; methods ; Electrophoresis, Gel, Pulsed-Field ; methods ; Lactobacillus ; classification ; isolation & purification ; Streptococcus thermophilus ; classification ; isolation & purification