Co-culture systems consisted of photosynthetic microorganisms and others heterotrophic microbes have attracted great attention in recent years. These systems show many advantages when compared with single culture grown under autotrophic conditions, such as less vulnerable to pollution and more stability, thus have been applied to wastewater treatment, soil remediation, biodegradable harmful substances, and production of high value-added products. In order to explore basic theory and further applications, we summarize here recent progresses in artificial co-culture systems of using photosynthetic microorganisms, to provide a current scientific understanding for the rational design of the co-culture system based on photosynthetic microorganisms using synthetic biology.
Coculture Techniques ; Heterotrophic Processes ; Microbiological Techniques ; trends ; Microbiota ; physiology ; Photosynthesis ; physiology ; Synthetic Biology ; trends

BACKGROUND/AIMS: Human gut microbiota harbors numerous metabolic properties essential for the host's health. Increased intestinal transit time affects a part of the population and is notably observed with human aging, which also corresponds to modifications of the gut microbiota. Thus we tested the metabolic and compositional changes of a human gut microbiota induced by an increased transit time simulated in vitro. METHODS: The in vitro system, Environmental Control System for Intestinal Microbiota, was used to simulate the environmental conditions of 3 different anatomical parts of the human colon in a continuous process. The retention times of the chemostat conditions were established to correspond to a typical transit time of 48 hours next increased to 96 hours. The bacterial communities, short chain fatty acids and metabolite fingerprints were determined. RESULTS: Increase of transit time resulted in a decrease of biomass and of diversity in the more distal compartments. Short chain fatty acid analyses and metabolite fingerprinting revealed increased activity corresponding to carbohydrate fermentation in the proximal compartments while protein fermentations were increased in the lower parts. CONCLUSIONS: This study provides the evidence that the increase of transit time, independently of other factors, affects the composition and metabolism of the gut microbiota. The transit time is one of the factors that explain some of the modifications seen in the gut microbiota of the elderly, as well as patients with slow transit time.
Aged ; Aging ; Biomass ; Colon* ; Constipation ; Dermatoglyphics ; Fatty Acids ; Fermentation ; Gastrointestinal Microbiome* ; Humans ; In Vitro Techniques* ; Metabolism* ; Microbiological Techniques

Anti-Bacterial Agents ; therapeutic use ; Bacterial Infections ; diagnosis ; drug therapy ; etiology ; microbiology ; Breast Implants ; microbiology ; Clinical Laboratory Techniques ; Early Diagnosis ; Humans ; Microbiological Techniques

This study aims at trying to establish a novel method of sterility test for injections based on biothermodynamics, in order to overcome the deficiencies of routine sterility tests such as long detecting cycle, low sensitivity and prone to misjudgments. A biothermodynamics method was adopted to rapidly detect the microorganism contamination of injections by monitoring the heat metabolism during the growth of microbe. The growth rate equal to or greater than zero and the heat power difference of P(i) and P(0) with three folds higher than the noise of baseline were chosen as indexes to study the heat change rule of microbe. In this way, the effectiveness of the new method to detect strains required by conventional sterility test or in injection samples was also investigated. Results showed that the Gram-positive bacteria, Gram-negative bacteria and fungi demanded by sterility testing methodology could be detected by biothermodynamics method within 10 hours, with the sensitivity lower than 100 CFU x mL(-1). Meanwhile, this method was successfully applied to the sterility test of Compound Yinchen injection (FFYC), Shuanghuanglian powder injection (SHL) and Compound Triamcinolone injection (TAND) which were sterilized with different degrees. Therefore, the biothermodynamics method, with advantages of fast detection and high sensitivity, could be a complementary solution for conventional sterility tests.
Anti-Inflammatory Agents ; administration & dosage ; chemistry ; Drug Contamination ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; Fungi ; isolation & purification ; Gram-Negative Bacteria ; isolation & purification ; Gram-Positive Bacteria ; isolation & purification ; Hot Temperature ; Injections ; Microbiological Techniques ; methods ; Sensitivity and Specificity ; Sterilization ; Triamcinolone ; administration & dosage ; chemistry

Heparinase II (Hep II) from Flavobacterium heparinum is an enzyme that could specifically cleave certain sequence of heparin and heparan sulfate. In this work, fermentation conditions of recombinant heparinase II (His-Hep II) producing bacteria were optimized, including initial induction time, inducer (IPTG) concentration, induction temperature and induction time. The optimum conditions were as follows: cultivating recombinant bacteria to exponential prophase under 37 degrees C, then adding IPTG to a final concentration of 0.3 g/L, finally cultivating recombinant bacteria under 20 degrees C for 10 h. The total crude enzyme activity reached 570 U/L. Based on these results, high cell density fermentation of recombinant bacteria was studied. The final OD600 could reach 98 and the total crude enzyme activity of His-Hep II increased to 9 436 U/L.
Fermentation ; Flavobacterium ; metabolism ; Microbiological Techniques ; Polysaccharide-Lyases ; biosynthesis ; Recombinant Proteins ; biosynthesis

OBJECTIVETo analyze and compare vaginal microbiomes in healthy women at child-bearing ages and patients with bacterial vaginosis (BV).

METHODSA total of 74 vaginal swabs of the vaginal fornix were collected from 37 BV patients and 37 healthy women. BV status was assessed according to Amsels clinical criteria for all the subjects and confirmed using Gram-stain criteria (Nugent scores). Genomic DNA of the samples was extracted for amplifying the 16S rRNA V6 hypervariable region by PCR and pyrosequencing by Illumina. BIPES, UCHIME, TSC and GAST were employed to analyze the information of the species from the samples.

RESULTSLactobacillus was the predominant species in healthy women (more than 95%), including mainly L. iners and L. crispatus, with a small quantity of Gardnerella, Granulicatella, Streptococcus, Prevotella, Escherichia and other genus. The α diversity was significantly increased in 30 BV patients (P<0.001), and β diversity also changed obviously shown by decreased Lactobacillus (varying from 45% to 1%, consisting mainly of L. iners) or even absence Lactobacillus in 6 cases, with increased relative abundance of Gardnerella, Prevotella, Granulicatella, Anaerococcus, Parvimonas, Peptoniphilus.harei, Peptostreptococcus, and Dialister. Different from previous data, 7 BV cases showed a predominance of the rare species L.gasseri and L.acidophilus (75% to 50%).

CONCLUSIONLactobacillus is the predominant vaginal species in healthy women (mainly L. iners and L. crispatus) co-existing with many other bacteria and a variety of microorganisms. Lactobacillus is significantly decreased and even absent in most of BV patients, and some cases show the predominance of the rare species L.gasseri and L.acidophilus.

Adult ; Case-Control Studies ; DNA, Bacterial ; genetics ; Female ; Humans ; Microbiological Techniques ; Microbiota ; Middle Aged ; RNA, Ribosomal, 16S ; genetics ; Reagent Kits, Diagnostic ; Vagina ; microbiology ; Vaginosis, Bacterial ; microbiology ; Young Adult

OBJECTIVETo compare the detection effect of Legionella pollution in spring water by three methods, namely traditional plating method, fluorescent quantitation PCR method and ethidium monoazide (EMA) fluorescent quantitation PCR method.

METHODSEvery month (except May), we collected 11 water samples from the 5 selected hot spring pools in one hot spring resort in Beijing in 2011. A total of 121 water samples were collected, and then were detected by the above three methods qualitatively and quantitatively.

RESULTSIn our study, the Legionella pollution rate was separately 74.4% (90/121), 100.0% (121/121) and 100.0% (121/121) by the above three methods. The quantitative value of Legionella in the 121 water samples detected by the three methods were around 0.10-216.00 colony-forming units (CFU)/ml, 1.47-1557.75 gene units (GU)/ml and 0.20-301.69 GU/ml, respectively. The median (25th and 75th percentiles) was 75.30 (32.51-192.10) GU/ml, 36.46 (16.08-91.21) GU/ml and 5.30 (0.00-33.70) CFU/ml, respectively. The difference in the quantitative value of Legionella detected by the three methods showed statistical significance (χ(2) = 187.900, P < 0.01). The quantitative value of Legionella detected by fluorescent quantitation PCR method was the highest, followed by the value Legionella detected by EMA-fluorescent quantitation PCR method and traditional plating method.

CONCLUSIONThe sensitivity of the PCR methods was higher than traditional plating method, in detecting Legionella pollution in spring water, especially the EMA- fluorescent quantitation PCR method, which was more suitable for detecting Legionella in water.

Environmental Monitoring ; methods ; Hot Springs ; microbiology ; Legionella ; classification ; isolation & purification ; Microbiological Techniques ; Water Microbiology

OBJECTIVETo apply pyrosequencing technique in the detection of the common pathogens in sepsis.

METHODSThe primers for amplification and sequencing in pyrosequencing were designed according to alignment of the bacterial 16S rRNA sequence. Bacterial genomic DNA was extracted for pyrosequencing, and the pathogen species were determined according to the sequencing data obtained.

RESULTSPyrosequencing effectively yielded the sequencing data of the 28 bp sequences of the pathogens and clearly distinguished the pathogen species of Streptococcus pyogenes, Streptococcus pneumonia, Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumonia, Neisseria meningitides, and Salmonella, but failed to distinguish Staphylococcus epidermidis from Staphylococcus aureus.

CONCLUSIONPyrosequencing technique can effectively distinguish the common pathogens in sepsis at the species level.

Bacteria ; classification ; isolation & purification ; DNA Primers ; DNA, Bacterial ; genetics ; Microbiological Techniques ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S ; Sepsis ; microbiology

OBJECTIVES: This study aimed to a) evaluate the knowledge of water refilling station (WRS) owners and operators regarding the proper techniques and procedures applicable to WRS based on the Certification Course for Water Refilling Station and Plant Operators (CCWRSPO); b) assess compliance to regular physical-chemical and microbiological testing of product water and sanitary permit acquisition and c) determine the quality of product water of selected water refilling stations (WRS) in a municipality in Cavite.

METHODS: The study includes WRS owners and operators who participated in the CCWRSPO from 2005 to 2009. A 50-item objective examination administered by the researchers was used to evaluate the knowledge of the respondents. This was formulated based on the objectives of the CCWRSPO. The compliance to legal requirements for WRS was assessed according to the results of the physical-chemical and microbiological tests (Multiple Tube Fermentation Technique and Pour Plate Method) and the presence of an updated sanitary permit. Results of product water analyses were compared to the 2007 Philippine National Standards for Drinking Water. Water refilling stations that failed to meet at least one of the three legal requirements were considered as "non-compliant".

RESULTS AND CONCLUSION: Results showed that 71.8% of the respondents passed the written examination whereas 28.2% obtained scores less than 50.0%. Chi-square analysis indicated that there was no significant difference between the knowledge of the trainees in 2005 to 2008 and the trainees in 2009. Similarly, majority (78.9%) of the WRS included in the study were found to be non-complaint with the provisions of P.D. 856 and the prescribed standards for water quality. The quality of product water served as an important determinant of the compliance of WRS. Although majority of the water samples tested had acceptable microbiological examination results, 16.9% of the samples exceeded the standards for microbiological water quality. Aside from this, the non-compliance of WRS was attributed to the absence of an updated sanitary permit, which was one of the important indicators of product water quality. Chi-square analysis showed that the trainees who have been operating WRS for only a year after the certification course were less compliant as compared to those operating for two to five years.

Water Quality ; Drinking Water ; Fermentation ; Patient Compliance ; Microbiological Techniques ; Surveys And Questionnaires ; Certification

Child ; Clinical Laboratory Techniques ; Humans ; Microbiological Techniques ; Viral Load ; Virus Cultivation ; Virus Diseases ; diagnosis