Fecal indicator bacteria diversity and decay in an estuarine mangrove ecosystem of the Xuan Thuy National Park, Vietnam.

Mangroves are complex and dynamic ecosystems that are highly dependent on diverse microbial activities. In this study, laboratory experiments and field studies for fecal indicator bacteria (FIB) decay rates are carried out for the first time in the Xuan Thuy Mangrove Forest Reserve of Vietnam. Results show that there are significant differences in bacterial diversity in the water of mangrove areas that have been deforested compared to those which have been planted. The highest mean total coliform (TC) and Escherichia coli (EC) values were found in the natural mangroves (3,807±2,922 and 964±1133 CFU 100 ml-1, respectively). The results indicated that the source of contamination and seasonal changes affect the abundance of fecal bacteria. These results were exceeding by far the safety guidelines for individual, non-commercial water supplies in most of the samples. In the planted mangrove sampling sites, the highest mean Fecal streptococci (FS) values of 1,520±1,652 CFU 100 ml-1 were found. Microbial die-off rates were calculated over 5 days, and observed to be systematically higher for TC than for EC.

Expression of recombinant T-cell epitopes of major Japanese cedar pollen allergens fused with cholera toxin B subunit in Escherichia coli.

Peptides containing T-cell epitopes from allergens, which are not reactive to allergen-specific IgE, are appropriate candidates as antigens for specific immunotherapy against allergies. To develop a vaccine that can be used in practical application to prevent and treat Japanese cedar pollen allergy, four major T-cell epitopes from the Cry j 1 antigen and six from the Cry j 2 antigen were selected to design cry j 1 epi and cry j 2 epi, DNA constructs encoding artificial polypeptides of the selected epitopes. To apply cholera toxin B subunit (CTB) as an adjuvant, cry j 1 epi and cry j 2 epi were linked and then fused to the CTB gene in tandem to construct a fusion gene, ctb-linker-cry j 1 epi- cry j 2 epi-flag. The fusion gene was introduced into a pET-28a(+) vector and expressed in Escherichia coli BL21(DE3). The expressed recombinant protein was purified by a His-tag affinity column and confirmed by western blot analysis using anti-CTB and anti-FLAG antibodies. The purified recombinant protein also proved to be antigenic against anti-Cry j 1 and anti-Cry j 2 antibodies. Expression of the recombinant protein induced with 1mM IPTG reached a maximum in 3-5h, and recovery of the affinity-purified recombinant protein was approximately 120mg/L of culture medium. The present study indicates that production of sufficient amounts of recombinant protein with antigenic epitopes may be possible by recombinant techniques using E. coli or other bacterial strains for protein expression.