The prevalence of antibiotic resistant bacteria in aquaculture has reached alarming proportions and intensified the search for microbe derived antimicrobial compounds. This study isolated bacteria from the intestine of Sagor catfish (Hexanematichthys sagor) and screened it for antagonistic properties. Five out of 334 bacterial isolates inhibited growth of fish pathogens. The 5 bacterial strains included relatives of Shewanella haliotis, Myroides odoratimimus, Vibrio harveyi, Vibrio alginolyticus and Alcaligenes faecalis. The growth profiles and probiotic properties of these bacteria were examined. The results showed that the isolate 9 (3) 220.127.116.11, whose closest relative was S. haliotis exhibited growth and probiotic advantage compared to the other bacterial strains, such as highest doubling time and the ability to survive at all experimental temperatures (18 to 60℃) , and bile concentrations (0.01 to 1.00%) and pH (pH2 to 9) . While the bacteria with probiotic properties were successfully isolated. Further study is necessary to examine the efficiency of the probiotic candidate bacteria in boosting fish immunity against pathogens.
Although the most common bacteria in the supragingival plaque are Gram-positive streptococci, no extensive investigations have been conducted into the susceptibility of these species to chlorhexidine and cetylpyridinium chloride. Therefore, in this study, we investigated the susceptibility of 80 streptococcal strains in planktonic or biofilm states to these two antimicrobial agents. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the planktonic streptococci were measured using the microdilution method, as were the minimum biofilm inhibitory concentration (MBIC) and minimum biofilm eradication concentration (MBEC) measured on streptococcal biofilms formed on 96-well plates. In all species, the MIC, MBC, MBIC, and MBEC values were higher for chlorhexidine than for cetylpyridinium chloride, with sensitivity values varying according to species. For chlorhexidine, the MIC, MBC, and MBIC values showed statistically significant differences among species. However, only MBEC values showed statistically significant differences for cetylpyridinium chloride. The MIC against Streptococcus mutans and the MBC against Streptococcus salivarius were significantly lower than those against the other species. With he exception of a few species, most of the bacterium susceptibility values were higher in the biofilm state than in the planktonic state.
Striped catfish (Pangasianodon hypophthalmus) farming in the Mekong Delta Vietnam (MKDVN) importantly contributes to national aquaculture export. Currently, however, diseases occur more frequently across the entire MKDVN region. One of the most common types is hemorrhagic septicemia caused by Aeromonas hydrophila. In this study, isolation and selection of the phages for control in vitro Aeromonas hydrophila were conducted. 24 phages were isolated from 100 striped catfish pond water samples. Next, lytic activity of these phages was clarified. Four phages with short latent period (about 25 to 40 min) and/or high burst size (about 67 to 94 PFU/ cell) were selected to evaluate their infection activity to different phage-resistant A. hydrophila strains. Two phages termed as TG25P and CT45P were subjected to the phage cocktail to inactivate A. hydrophila. Re-growth of the host bacteria appeared about eight hours after treatment. Usage of the phage cocktail that attach different host bacterial receptors is not always much effective than usage of single phage. This is the first report about phage therapy to control A. hydrophila isolated from striped catfish. Some challenges in the phage cocktail were shown to achieve strategies in prospective studies in the context of high antibiotic resistance of A. hydrophila.
In this study, we developed a system, known as MicroStarTM Rapid Microbe Detection System (RMDS) , to detect Lactobacillus brevis, which usually requires 2-4 days for examination by the conventional plate count procedure, for beer quality control using a bioluminescence method within 24 hr and also aimed to develop a technology to detect bacterial growth without the need for cultivation. We used a highly sensitive luminous reagent that increased the activity of the luciferin- luciferase reaction to 2.5×10－18 mol ATP/0.2 μl and could detect even a single lactic acid bacterial cell. The limitation of the method was that ATP derived from the beer hindered bacterial measurement and the supply of energy source to secure ATP of lactic acid bacterial cell. The sample beer was filtered through a membrane filter, avoiding the formation of beer foam to the best extent, the filter was cleaned with 10% ethanol and 0.1% sodium hydrogen carbonate solution, and incubated on a GMY agar plate (1% glucose, 0.2% malic acid, 0.67% yeast nitrogen base, 1% agar; pH 5.2) at room temperature for 2 hr. Post incubation of the filter, bacterial cell count was measured with RMDS. This method could overcome the hindrance of ATP measurement and could stably detect lactic acid bacteria without the need for cultivation.
Legionella spp. exist naturally in association with amoeba in water environments and are known to be the etiological agent of a severe form of pneumonia. To detect diverse Legionella populations in cooling tower water systems, amoebic coculturing was performed for 15 water samples obtained from five different kinds of facilities in six geographically different locations. The growth of Legionella in coculture with Acanthamoeba sp. cells was monitored by quantitative PCR targeting Legionella-specific 16S rRNA genes. Seven out of the 15 samples were positive for Legionella growth and subjected to clone library analysis. A total of 333 clones were classified into 14 operational taxonomic units composed of seven known species and seven previously undescribed groups. Four of the seven Legionella-growth-positive samples harbored detectable levels of free-living amoeba and were predominated by either L. drozanskii or L. lytica, by both L. bozemanii and L. longbeachae, or by a not-yet-described group named OTU 4. The Legionella-growth- positive samples contained higher ATP levels (>980 pM) than the growth-negative samples (<160 pM) , suggesting that ATP content would be a good indicator of the presence of viable but nonculturable Legionella populations able to grow with amoeba.
A useful tool for the screening of fungi producing biologically active secondary metabolites such as antibiotics and cytotoxic substances has been developed. An agar plate-organic solvent interface cultivation (A/S-IFC) system, which comprised a hydrophobic organic solvent (upper phase) , a fungal mat (middle phase) and an agar plate (lower phase) , was constructed. The metabolite profiles were compared among the A/S-IFC, a traditional submerged cultivation (SmC) and an extractive liquid surface immobilization (Ext-LSI) system consisted of a hydrophobic solvent (upper phase) , a fungal cells–ballooned microspheres (middle phase) and a liquid medium (lower phase) , with high-performance liquid chromatography-photodiode array detector (HPLC-PDA) . In the A/S-IFC, many hydrophobic metabolites vastly different from those in the SmC were accumulated in the organic phase as with the Ext-LSI. For example, a valuable azaphilone, sclerotiorin, was remarkably produced into the organic phase in the A/S-IFC. The A/S-IFC was applied to the screening of antibiotic-producing fungi. As a result of paper disk method, it was found that 321 isolated among 811 strains produced antifungal metabolites (hit rate, 39.6%) . Furthermore, 8, 23, and 30 strains also produced cytotoxic metabolites against SKOV-3 (human ovary adenocarcinoma) , MESO-1 (human malignant pleural mesothelioma) , and Jurkat cells (immortalized human T lymphocyte) .
Effective spatial disinfection systems are required for human health care, public hygiene, and food and medicine manufacturing. Although some aerosolized disinfectants were already applied to its purpose, accurate evaluation systems were under constructed. In this study, the spatial sporicidal activity of aerosolized hypochlorite solution (AHS) to dormant cells, Bacillus subtilis spores, was evaluated by an originally designed chamber system. In the test-chamber, AHS was supplied and existed as micro-droplets, and environmental relative humidity (RH) could be controlled. Available chlorine (AC) exposure was also controlled with appropriate AC loading but was influenced by the acidity of AHS. Our results indicated that inactivation of spore was depend on AC exposure amount and time. On the other hand, unsaturated environmental RH markedly decreased spore inactivation. This study indicated that our test-chamber system can provide reproducible test data under a homogeneous air condition, and, thereby, that the data obtained by the chamber system should contribute to predicting the AC-required dose to disinfect a whole building.
The effect of food preservatives and sanitizers at low concentrations on the induction of Escherichia coli into a viable but nonculturable (VBNC) state was investigated. When E. coli was incubated in physiological saline at 37℃, the viable cell count measured by plate counting was approximately 3-logs lower than that measured by flow cytometry after 30 days. This difference, and morphological changes in cells, confirmed the transition of E. coli into a VBNC state. Adding 10 μg/l of sorbic acid significantly promoted the induction of E. coli into a VBNC state. This effect was not seen with benzoic acid or sodium hypochlorite at the same concentration. Resuscitation of E. coli VBNC cells was successful when they were grown in nutrient broth containing sodium pyruvate. These results suggest that the presence of low concentrations of food additives in a food manufacturing environment may act as potential triggers for bacterial VBNC induction.