Three fungal species were evaluated for their abilities to saccharify pure cellulose. The three species chosen represented three major wood-rot molds; brown rot (Gloeophyllum trabeum), white rot (Phanerochaete chrysosporium) and soft rot (Trichoderma reesei). After solid state fermentation of the fungi on the filter paper for four days, the saccharified cellulose was then fermented to ethanol by using Saccharomyces cerevisiae. The efficiency of the fungal species in saccharifying the filter paper was compared against a low dose (25 FPU/g cellulose) of a commercial cellulase. Total sugar, cellobiose and glucose were monitored during the fermentation period, along with ethanol, acetic acid and lactic acid. Results indicated that the most efficient fungal species in saccharifying the filter paper was T. reesei with 5.13 g/100 g filter paper of ethanol being produced at days 5, followed by P. chrysosporium at 1.79 g/100 g filter paper. No ethanol was detected for the filter paper treated with G. trabeum throughout the five day fermentation stage. Acetic acid was only produced in the sample treated with T. reesei and the commercial enzyme, with concentration 0.95 and 2.57 g/100 g filter paper, respectively at day 5. Lactic acid production was not detected for all the fungal treated filter paper after day 5. Our study indicated that there is potential in utilizing in situ enzymatic saccharification of biomass by using T. reesei and P. chrysosporium that may lead to an economical simultaneous saccharification and fermentation process for the production of fuel ethanol.

Aims: Oleaginous yeasts are widely used for the production of biodiesel feedstocks because of their high lipid content. This research was aimed to conduct random mutagenesis of Rhodotorula mucilaginosa using ethyl methane sulfonate (EMS) and identify the mutants with improved lipid production. Methodology and results: A total of twenty-two mutant isolates prescreened with cerulenin were produced and further characterized via M13 PCR fingerprinting to determine their polymorphism and genetic distances. Eight strains, namely M1, M2, M3, M4, M7, M10, M11 and M18, were chosen based on their genetic distances from the parental strain for biomass production. Six mutants (M1, M2, M3, M4, M7 and M18) showing the highest dry cell weights were further selected for evaluation of lipid production in a laboratory-scale bioreactor using glucose as a carbon source. Results indicated that parental strain exhibited lipid content of 1.83 g/L, while strains M1, M2, M3, M7 and M18 generated 2.37 g/L, 2.27 g/L, 2.27 g/L, 3.10 g/L and 3.83 g/L of intracellular lipid, respectively. These five mutants were identified to have significant increase in lipid production compared to the parental strain. Conclusion, significance and impact of study This study demonstrated enhanced lipid production in R. mucilaginosa by random mutagenesis. New generated strains had higher lipid productivity compared to parental strain and application of these strains in industry may reduce the overall cost of biodiesel production.

Thirty one Vibrio cholera isolates recovered from cholera outbreak in Bintulu, Sarawak (Malaysia) were detected with the presence of ctx gene by using specific PCR. These isolates were further characterized and differentiated by using the Enterobacterial Repetitive Intergenic Consensus PCR (ERIC-PCR) and BOX-PCR to determine their genomic fingerprints. The specific PCR result confirmed the identities of 27 isolates out of 31 as pathogenic V. cholerae. The ERIC-PCR generated several genetic profiles consisting of 4-6 bands with sizes in the range of 100 to 600 bp, while the BOX-PCR produced profiles numbering 2-7 bands in the sizes between 200 to 1000 bp. Based on the dendrogram generated from the DNA fingerprinting profiles (ERIC-PCR and BOX-PCR), all of the isolates can be divided into 2 main clusters that is further divided into 2 sub-clusters. The low genetic diversity of the isolates indicated the outbreak of V. cholerae in the study area was due to the contamination from a single or few sources of V. cholerae.
Cholera

Aims: Bacillus cereus is a Gram-positive, rod-shaped and spore-forming bacterium. It is a ubiquitous bacterium which is widely distributed in several environments such as soil and plants and is commonly isolated from food and its processing environment. This study was aimed to determine the genetic diversity and antibiotic resistance of B. cereus isolated from sago processing in Sarawak. Methodology and results: Out of 120 samples, 42 B. cereus isolates were detected with the presence of hly gene of B. cereus by using specific polymerase chain reaction (PCR). Twenty B. cereus isolates were randomly selected and further characterized by pulsed-field gel electrophoresis (PFGE) of chromosomal DNA digested with NotI to examine the genetic diversity. The result of the PFGE analysis confirmed that the B. cereus strains in sago processing were genetically diverse. Based on the dendrogram generated, B. cereus strains were grouped into two major clusters and these clusters were grouped together based on sources of isolation. The investigation on the antibiotic resistance of B. cereus strains revealed that the B. cereus strains were uniformly highly resistant to penicillin and ampicillin and highly susceptible to imipenem and norfloxacin. Conclusion, significance and impact of study The results of this study suggest that the B. cereus isolated from sago processing derived from a mixture of sensitive and resistant strains with diverse genetic contents.