Probiotic characterization of lactic acid bacteria isolated from infants feces and its application for the expression of green fluorescent protein

Aims: In this study, lactic acid bacteria (LAB) were isolated from 42 healthy infants and determined for probiotic properties. Twelve LAB isolates with potential probiotic properties were selected and screened for their feasibility of heterologous protein expression by selection of erythromycin sensitive isolates. Methodology and results: One of eleven erythromycin-sensitive LAB isolates identified and designated as Lactobacillus fermentum 47-7 was able to acquire and stable maintain the Escherichia coli-Lactobacillus shuttle vector, pRCEID-LC13.9. Further electrotransformation of L. fermentum 47-7 with the recombinant pLC13.9:LDH-PRO1:GFPuv containing green fluorescent protein (GFP) gene found that recombinant L. fermentum can express GFP. Conclusion, significance and impact of study: The probiotic L. fermentum isolate can be used as host for expression of heterologous proteins and could possibly be further developed as the alternate oral delivery system for various biomolecules for biotechnological application.

Detection and Differentiation of Mycobacterium Species in Clinical Specimens by Real-Time Polymerase Chain Reaction

Mycobacterium tuberculosis complex and Mycobacterium avium complex are causative agents of tuberculosis which is a major cause of human morbidity and mortality. Rapid diagnosis and species differentiation of mycobactria in specimens are important to control the disease and use appropriate antimicrobial therapy. In this study, primers and probe were design based on 16S-23S rDNA spacer sequence by using LightCycler 2.0 software. One set of primer, Rank 9, was used to perform real-time PCR and conventional PCR for detection and differentiation of Mycobacterium spp. Sybr Green dye for signal detection, successfully amplified all extracted DNA of mycobacteria. Its amplicon can be visualized by gel electrophoresis with the corresponding length of 214 bp. The nonspecific products of about 300 bp, were found in conventional PCR. The result suggested that these primers are not appropriate for conventional PCR. A hybridization probe for strain differentiation and verification of the protocol in clinical specimens is needed to complete the protocol.

In-vitro evaluation of antiseptics and disinfectants against agianst Burkholderia pseudomallei from clinical isolates.

Objective: Chemical widely used as antiseptics and disinfectants for killing various microorganisms in many studies whether no evaluation of chemical agents against B. pseudomallei . Materials: Our study performed  various concentrations of common used as antiseptics and disinfectants in hospitals for example phenol derivatives, alcohol, halogen compounds, aldehyde and quaternary ammonium compounds against B. pseudomallei. These informations will help physician who work in infectious unit easy choose these agents for killing or controlling spread in hospital environment especially  these organisms can survive in water or humid area as Psedomonas aeruginosa. Results: This study using chemical agents in clinic performed in-vitro evaluation on B. pseudomallei from various specimens of Srinagarind hospital by dilution method.  This found that good antiseptics against these organisms were 70%Alcohol 2%ProvidineÒ 0.1%ThimersalÒ and 2% MecurochromeÒ; good disinfectants against that were 4%Formaldehyde, 2%LysolÒ and 3%H2O2. The 0.5% HibitaneÒ  has modertaedly  antibactericidal effect on B. pseudomallei . Both 1:100 SavlonÒ and 0.1%AcriflavinÒ were poor antibactericidal against these organisms. Whereas 1% VirkonÒ  was optimum concentration for killing these organisms and against contaminations not more than 107 cells/ml. Conclusions: The effective chemical agents against  B. pseudomallei  were  0.1-0.5% Chlorine, 4%Foramldehyde, 3%H2O2, 2%LysolÒ, 70%Alcohol, 2%ProvidineÒ, 0.1%ThimerosalÒ and 2% MecurochromeÒKeywords: antiseptics, disinfectants, Burkholderia pseudomallei, clinical isolates