Optimization of Salmonella Typhi biofilm assay on polypropylene microtiter plates using response surface methodology

The objective of this study was to develop an optimized assay for Salmonella Typhi biofilm that mimics the environment of the gallbladder as an experimental model for chronic typhoid fever. Multi-factorial assays are difficult to optimize using traditional one-factor-at-a-time optimization methods. Response surface methodology (RSM) was used to optimize six key variables involved in S. Typhi biofilm formation on cholesterol-coated polypropylene 96-well microtiter plates. The results showed that bile (1.22%), glucose (2%), cholesterol (0.05%) and potassium chloride (0.25%) were critical factors affecting the amount of biofilm produced, but agitation (275 rpm) and sodium chloride (0.5%) had antagonistic effects on each other. Under these optimum conditions the maximum OD reading for biofilm formation was 3.4 (λ600 nm), and the coefficients of variation for intra-plate and inter-plate assays were 3% (n = 20) and 5% (n = 8), respectively. These results showed that RSM is an effective approach for biofilm assay optimization

Production of hydrogen energy from dilute acid-hydrolyzed palm oil mill effluent in dark fermentation using an empirical model

Hydrogen generation was studied using palm oil mill effluent (POME) as an agro-industrial waste obtained from the palm oil industry. POME was subjected to a dilute acid hydrolysis step by HCl (37% v/v) to release fermentable sugars from cellulosic content. POME hydrolysate obtained was used as a substrate for hydrogen generation. The composition of POME hydrolysate showed glucose and xylose were the main monomeric sugars liberated. Hydrogen production was performed in dark fermentation process, in which the new bacterial strain Clostridium acetobutylicum YM1 was cultivated on POME hydrolysate based on a central composite design (CCD). CCD was constructed by considering three pivotal process variables including incubation temperature, initial pH of culture medium and microbial inoculum size. An empirical model, namely second-order polynomial regression model was generated and adjusted to CCD data. The analysis of empirical model generated showed that the linear and quadratic terms of temperature had a highly significant effect on hydrogen generation (P < 0.01). Furthermore, the quadratic effects of initial pH value of culture medium and inoculum size had a significant effect on hydrogen production at 95% probability level (P < 0.05). The regression model also showed that the interaction effect between temperature and initial pH value of the culture medium on the hydrogen generation was highly significant (P < 0.01). The empirical model suggested that the optimum conditions for hydrogen production were an incubation temperature of 38 °C, initial pH value of 5.85 and inoculum size of 17.61% with predicting the production of a cumulative hydrogen volume of 334.2 ml under optimum conditions. In order to validate the optimum conditions determined, C. acetobutylicum YM1 was cultivated on POME hydrolysate in optimum conditions. Verification test results showed that a cumulative hydrogen volume of 333.5 ml and a hydrogen yield of 108.35 ml H2/g total reducing sugars consumed were produced

Utilization of palm kernel cake as a renewable feedstock for fermentative hydrogen production

Fermentative hydrogen generation was studied using palm kernel cake (PKC) as sustainable cellulosic biomass. PKC was subjected to an acid hydrolysis approach using dilute H2SO4 (7% v/v). PKC hydrolysate obtained was then diluted (70%) and used as a substrate for hydrogen generation. Chemical analysis showed that the main fermentable sugars in diluted PKC hydrolysate were glucose, xylose and mannose with the concentrations of 2.75 g/L, 2.60 g/L and 27.75 g/L, respectively. Hydrogen production was carried out by the cultivation of Clostridium acetobutylicum YM1 on PKC hydrolysate. The effect of incubation temperature, the initial pH of culture medium and microbial inoculum size on hydrogen production was studied using a statistical model. The analysis of the model generated showed that the initial pH value of the culture medium and inoculum size had significant effects on the hydrogen production. The study showed that the optimum conditions for the biohydrogen production were 30.57 °C temperature, pH 5.5 and 20% inoculum size. A verification experiment was performed in the optimum conditions determined. Experimental results of the verification test showed that a cumulative hydrogen volume of 1575 ml/L was generated with consuming 2.75 g/L glucose, 2.20 g/L xylose and 16.31 g/L mannose