Poultry meat can be contaminated by different types of microorganisms during processing in processing plant. The microbiological quality of chicken carcasses and along with processing steps and environmental condition was analyzed in this study in an ISO 22000:2005 certified poultry processing plant of Kathmandu. Standard plate count method was applied for the enumeration and detection of total mesophilic bacteria, total coliform, total faecal coliform, Staphylococcus load along with selected pathogens like Salmonella spp., S. aureus, Escherichia coli, Clostridium perfringens, and Listeria spp. in chicken meat at four processing step (evisceration, final washing, frozen and market). It was observed that the level of microbial load decreased with subsequent processing phases in poultry processing plant where high level of bacteria were reduced during final washing and frozen phase. After processing poultry meat in an ISO 22000:2005 certified meat processing plant, total aerobic mesophilic count, total coliform count, total faecal coliform count, total Staphylococcus count were decreased from 6.92 to 4.45 log CFU/g, 3.49 to 2.19 log CFU/g, 2.41 to nil log CFU/g, and 3..43 to 1.99 log CFU/g respectively. Pathogenic bacteria like Salmonella spp., C. perfringens, and Listeria spp. were absent in chicken meat at the fourth processing step. Prevalence of E. coli was reduced from 37.4% to 10.2%, whereas S. aureus was decreased from 18.57% to 17.1%. It was concluded that the final washing and freezing steps were the Critical Control Point (CCP) to control microbial hazards in poultry processing phase.

Pectinase are the group of enzymes that catalyze the degradation of pectin substances through de-polymerisation and de-esterification. This study is concerned on the isolation, screening and selection of suitable strain of pectinolytic organism and optimization of cultural conditions for the biosynthesis of pectinase. From the soil samples collected from Lalitpur, Kathmandu, Gulmi, Manang and Dang, 18 fungal colonies were isolated on the basis of halozone formation on Potato dextrose agar and identified. Enzyme production was carried out by submerged state fermentation. The partially purified enzymes showing higher pectinolytic activity were characterized on the basis of temperature of incubation, substrate concentration and pH of the substrate by Dinitro salicylic acid assay (DNSA) method. The fungal isolate showing highest enzyme activity was subjected to optimization of culture medium for the production of enzymes. On optimization, it was found that MG1 (Aspergillus niger) was the most potent strain at 1.5% substrate (pectin) concentration, pH 4.5 and temperature of 30°C. On the enzyme application, the yield of the orange juice, Total Soluble Solid and absorbance increased as the concentration of the enzyme increased and hence increasing the possibility of being used commercially for maximum pectinase production. DOI: http://dx.doi.org/10.3126/jfstn.v8i0.11752 J. Food Sci. Technol. Nepal, Vol. 8 (65-70), 2013

Amylases are starch degrading enzymes which are produced by plants, animals and microorganisms. Amylases produced by microorganisms have a wide range of industrial applications such as in pharmaceutical, food, textile and paper industries. However, there are still limitations in the isolation of amylase producing microorganisms. The objective of this study was to isolate the potent amylase producing Bacillus sp. from soil samples and evaluate their abilities for inhibiting the aflatoxin producing Aspergillus flavus. In this study, 30 soil samples were used. For the screening and identification of Bacillus strain, morphological and biochemical tests were performed. Iodine assay was done to screen the potent amylase producers. Two parameters (pH and temperature) were used to optimize the cultural conditions for the production of amylase. To determine the total reducing sugar, dinitrosalicylic acid (DNS) assay was used. Altogether 29 colonies were selected and identified as Bacillus spp out of which 16 were selected to determine enzyme activity by cup plate method. Four isolates (DK9, DK10, IM4 and KD7) showing highest amylolytic activities (16 mm, 12 mm, 14 mm and 14 mm zone of hydrolysis) were subjected for further study. Isolate KD7 showed the highest amylolytic activity (0.19 U/mL) compared to other isolates. Maximum amylase production was found at pH 6 and temperature 50° C (0.19 U/mL). Among these 4 isolates, DK9 and KD9 showed strong antagonistic activity against Aspergillus flavus while DK10 and IM4 showed moderate antifungal activities. Thus, the bacterial isolate KD7 was identified as the most potent strain for maximum amylase production.

Background: Pseudomonas aeruginosa is an opportunistic human pathogen and are reported to cause acute and chronic infectious diseases. Due to its high ability to acquire resistance to many antibiotics, it has become a global public health threat. It consists of some virulence genes that may lead to its pathogenicity. The main objective of this cross-sectional study was to detect the virulence genes and antibiotic susceptibility pattern of P. aeruginosa isolated from clinical specimens collected from governmental hospital of Nepal.Methods: A total of 7898 clinical specimens were analyzed for the period of six months from November 2018 to April 2019. The specimens were cultured on Nutrient agar, Blood agar, MacConkey agar, Chocolate agar, Cysteine-Lactose, Electrolyte Deficient agar plates and were incubated at 37°C for 24 hours. All the isolates were identified by standard biochemical tests and further confirmed by growth on Cetrimide agar plate. The antibiotic susceptibility testing was performed by modified Kirby-Bauer disc diffusion method following CLSI guideline. Multiplex-PCR was done to detect the virulence genes oprL and toxA. Statistical analysis was carried out using IBM SPSS Statistic ver. 25 and the p-value was calculated at significance level (0.05%) by using Chi square.Results: Out of these specimens investigated, 87 isolates were tentatively identified to be P. aeruginosa in which 20 (22.98 %) were found to be multidrug resistant. Comparatively, most of the P. aeruginosa were isolated from outpatients 63 (72.41 %) than inpatients 24 (27.58 %), from male 56 (64.36 %) than female 31 (35.63 %) and in age group 60-79 years (41.37 %). AST result showed the highest resistance of 100% with cefixime whereas susceptibilities of 83.9% and 81.6% with polymixin B and tobramycin were noticed respectively. The PCR results showed that all P. aeruginosa isolates carried oprL 87 (100%) and 83 (95.4 %) isolates showed toxA genes. Conclusion: The studies revealed that almost all P. aeruginosa harbors both oprL and toxA genes.

Objectives: The aim of this study was to determine the prevalence of methicillin resistant Staphylococcus aureus (MRSA) and antibiotic resistance pattern of the isolates from wound infections. Methods: A total of 706 wound specimens including pus and wound swab were analyzed in the laboratory of B and B Hospital, Lalitpur from May to October 2014. The specimens were cultured on Blood Agar and Mannitol Salt Agar plates and incubated at 37°C for 24 hours. Antibiotic susceptibility test was performed by modified Kirby-Bauer disc diffusion method. Strains resistant to cefoxitin (30mcg) with inhibition zone ≤ 21mm were identified as MRSA. Results: Out of 366 bacterial isolates, 90 (24.6%) were S. aureus and among them 16.7% were MRSA and 54.4% multi-drug resistant (MDR). All isolates were sensitive to vancomycin and most of the isolates were sensitive to cefoxitin (83.3%). High rate of resistance was observed towards penicillin (98.9%) and ampicillin (86.7%). All MRSA isolates and 52.9% of methicillin sensitive S. aureus (MSSA) were MDR. Conclusion: MRSA incidence is increasing in the population, and therapeutic measures are few and accompanied by diverse side effects. It is noteworthy to state that vancomycin is still the first line drug although vancomycin-resistant strains have been reported.