Potential surface active agent production using very low grade and cheap substrate by Bacillus subtilis as microbial cell factory

Bio-surfactants are surface-active molecules which are produced by the wide range of microbes including bacteria, fungi, moulds, and yeast. This study was conducted to identify bio-surfactants by Bacillus subtilis combined with use of cheap substrates and industrial wastes (Mustard cake, Whey and Soya cake) which are found locally in Nepal. Bacillus subtilis, one of the most potential bio-surfactants producer; was isolated from soil sample of hydrocarbon contaminated site. Isolates were grown in a Minimal Salt Media (MSM) with 10% (v/v) mustard oil cake, whey and soya cake separately. The presence and potential of surfactant was determined by the oil spreading technique, emulsification index (%E24) and surface tension measurement. It was revealed that the surface tensions of cell free extract were 54.41, 60.02 and 56.64 mN/m for from mustard cake, whey and soya cake respectively as compared to distilled water (72.09) at 25oC. The emulsification index values was found to be highest in engine oil from the bio-surfactant extracted from mustard cake, soya cake and whey respectively. Similarly, mustard oil showed the lowest value of emulsification index. The highest emulsification activity was shown in mustard oil i.e. 1.13 from the cell free extract from mustard oil and lowest in engine oil i.e., 0.07, by the extract from soya cake medium, when measured in spectrophotometer at 540 nm. In conclusion, strain of Bacillus subtilis was found to be the potential surface active agent producers on the mustard oil cake, which can be useful medium for various environmental, food, medicinal and industrial processes

Potential surface active agent production using very low grade and cheap substrate by Bacillus subtilis as microbial cell factory

Bio-surfactants are surface-active molecules which are produced by the wide range of microbes including bacteria, fungi, moulds, and yeast. This study was conducted to identify bio-surfactants by Bacillus subtilis combined with use of cheap substrates and industrial wastes (Mustard cake, Whey and Soya cake) which are found locally in Nepal. Bacillus subtilis, one of the most potential bio-surfactants producer; was isolated from soil sample of hydrocarbon contaminated site. Isolates were grown in a Minimal Salt Media (MSM) with 10% (v/v) mustard oil cake, whey and soya cake separately. The presence and potential of surfactant was determined by the oil spreading technique, emulsification index (%E24) and surface tension measurement. It was revealed that the surface tensions of cell free extract were 54.41, 60.02 and 56.64 mN/m for from mustard cake, whey and soya cake respectively as compared to distilled water (72.09) at 25oC. The emulsification index values was found to be highest in engine oil from the bio-surfactant extracted from mustard cake, soya cake and whey respectively. Similarly, mustard oil showed the lowest value of emulsification index. The highest emulsification activity was shown in mustard oil i.e. 1.13 from the cell free extract from mustard oil and lowest in engine oil i.e., 0.07, by the extract from soya cake medium, when measured in spectrophotometer at 540 nm. In conclusion, strain of Bacillus subtilis was found to be the potential surface active agent producers on the mustard oil cake, which can be useful medium for various environmental, food, medicinal and industrial processes

Population-level risks of alcohol consumption by amount, geography, age, sex, and year: a systematic analysis for the Global Burden of Disease Study 2020

BACKGROUND: The health risks associated with moderate alcohol consumption continue to be debated. Small amounts of alcohol might lower the risk of some health outcomes but increase the risk of others, suggesting that the overall risk depends, in part, on background disease rates, which vary by region, age, sex, and year. METHODS: For this analysis, we constructed burden-weighted dose-response relative risk curves across 22 health outcomes to estimate the theoretical minimum risk exposure level (TMREL) and non-drinker equivalence (NDE), the consumption level at which the health risk is equivalent to that of a non-drinker, using disease rates from the Global Burden of Diseases, Injuries, and Risk Factors Study (GBD) 2020 for 21 regions, including 204 countries and territories, by 5-year age group, sex, and year for individuals aged 15-95 years and older from 1990 to 2020. Based on the NDE, we quantified the population consuming harmful amounts of alcohol. FINDINGS: The burden-weighted relative risk curves for alcohol use varied by region and age. Among individuals aged 15-39 years in 2020, the TMREL varied between 0 (95% uncertainty interval 0-0) and 0·603 (0·400-1·00) standard drinks per day, and the NDE varied between 0·002 (0-0) and 1·75 (0·698-4·30) standard drinks per day. Among individuals aged 40 years and older, the burden-weighted relative risk curve was J-shaped for all regions, with a 2020 TMREL that ranged from 0·114 (0-0·403) to 1·87 (0·500-3·30) standard drinks per day and an NDE that ranged between 0·193 (0-0·900) and 6·94 (3·40-8·30) standard drinks per day. Among individuals consuming harmful amounts of alcohol in 2020, 59·1% (54·3-65·4) were aged 15-39 years and 76·9% (73·0-81·3) were male. INTERPRETATION: There is strong evidence to support recommendations on alcohol consumption varying by age and location. Stronger interventions, particularly those tailored towards younger individuals, are needed to reduce the substantial global health loss attributable to alcohol. FUNDING: Bill & Melinda Gates Foundation

Antibiotic resistance patterns of Staphylococcus aureus, Escherichia coli, Salmonella, Shigella and Vibrio isolated from chicken, pork, buffalo and goat meat in eastern Nepal

Abstract Objective Food-borne pathogens are a major cause of illnesses, death and expenses. Their occurrence in meat and other food is considered a global health problem. The burden of food-borne disease is increasing due to antimicrobial resistance which represents a greater risk of treatment failure. However, very little is known about the antibiotic resistance profile of food-borne pathogens in Nepal. This study was conducted to examine the antibiotic resistance profile of common food-borne bacterial pathogens isolated from raw meat sold in Nepal. A total of 83 meat samples were collected from the market and analyzed. Results The prevalence of Staphylococcus aureus, Escherichia coli, Salmonella, Shigella, and Vibrio were 68%, 53%, 35%, 6%, and 6% respectively. The resistance of Salmonella was most frequently observed to amoxicillin (100%), tetracycline (24%), chloramphenicol (11%), and nalidixic acid (11%). S. aureus was resistant to amoxicillin (100%) followed by tetracycline (63%), nalidixic acid (17%), and cefotaxime (13%) respectively. Vibrio isolates resisted amoxicillin (100%), tetracycline (40%) and chloramphenicol (20%). Shigella expressed the highest resistance to amoxicillin (100%), followed by chloramphenicol (80%), tetracycline (60%) and nalidixic acid (20%). E. coli exhibited the highest resistance to amoxicillin (100%), followed by tetracycline (93%), nalidixic acid (25%) and cefotaxime (19%). </jats:sec

Interaction of Cetylpyridinium Chloride and Methylene Blue in Methanol-Water Solvent Media

The interaction of Methylene blue with Cetylpyridinium chloride (CPC) in a series of solvents containing 0.1, 0.2 and 0.3 volume fractions of methanol in water were studied at room temp (300±2 K) by UV-Vis spectrophotometer. The CPC concentrations were varied from 0.2 x 10-3 to 2 x 10-3 mol dm-3. A fixed dye concentration of 2.5 x 10-5 mol dm-3 was used for this purpose. The spectrum analysis was performed for both the dye solution and mixtures containing dye and surfactant of different concentrations. The dye solution showed the absorbance maximum at 664 nm and a shoulder at 610 nm. In 0.3 volume fraction of methanol as solvent, the dye and surfactant mixed solution showed a shift in peak values towards shorter wavelength (blue shift) from peak of the dye solution. The shift in peak increased with increase in surfactant concentration up to 0.6 x10-3 mol dm-3, beyond which no further shift increment occurred. In the case of 0.1 and 0.2 volume fractions of methanol in water, no shift in peak was found at all concentrations of dye and surfactant. The absorbance of dye and surfactant mixed solution decreased gradually in 0.2 volume fraction of methanol with increase in surfactant concentration, whereas no significant change in absorbance was seen in 0.1 volume fraction of methanol. The shift decreased gradually with decrease in methanol concentration in the solvent.</jats:p

Antibiogram Profiling and Thermal Inactivation of Staphylococcus aureus and Escherichia coli Isolated from Milk of Dharan, Nepal

Milk is an excellent medium for the growth of many bacteria. This study aimed to determine antibiotic profiling and thermal inactivation of Staphylococcus aureus and Escherichia coli isolated from raw milk of Dharan. Total viable count, total Staphylococcal count, and total coliform count were carried out by conventional microbiological methods. Identification was done on the basis of Gram staining and biochemical tests. The antibiotic susceptibility test of the isolates carried out by the modified Kirby-Baur disc diffusion method. Thermal inactivation of S. aureus and E. coli were carried out by subjecting to thermal treatment in a water bath. Total plate count ranged from 204×104 CFU/mL to 332×105 CFU/mL. Total staphylococcal count and total coliform count ranged from 14×105 CFU/mL to 8×106 CFU/mL and 11×104 CFU/mL to 3×106 CFU/mL respectively. S. aureus showed an increasing resistance patterns towards Ampicillin, Cefotixin, Carbenicillin and Cefotaxime. Ciprofloxacin, Erythromycin, Amikacin, Gentamycin, Azithromycin, and Chloramphenicol were found to be effective against S. aureus. All the E. coli isolates were resistant to Ampicillin and least resistant to Cefotixin. Chloramphenicol, Amikacin, Azithromycin, and Nalidixic acid were found highly effective to E. coli. The D-values for S. aureus at 56°C, 58°C and 60°C were 1.36 min, 1.19 min, and 1.09 min respectively. The Z-value was 14.92°C. While D-values were obtained as 0.98 min, 0.75 min, and 0.57 min for E. coli at 56° C, 58° C and 60° C respectively, and Z-value was 9.75° C. Hence, S. aureus was found to be more heat resistant than E. coli.</jats:p

Antibiogram of Biofilm Producing and Non-Producing Community Acquired-Methicillin Resistant Staphylococcus aureus Isolated from Potential Risk Population of Dharan, Nepal

Abstract Background Staphylococcus aureus is one of the common cause of hospital acquired infection and community acquired infections. Nowadays these organisms became resistant towards variety of drugs. MRSA is the emerging antibiotic resistant bacteria that are resistant to methicillin antibiotic and known to be the infectious pathogen causing severe infection and a cause of fatal mortality.Materials and methodology Altogether 200 nasal swabs and 200 hand swabs were obtained from participants and transported to microbiology lab in cold chain. The samples were swabbed in mannitol salt agar (MSA) containing oxacillin powder of 6mg/L and incubated at 37°C for 24 hrs. Staphylococcus aureus colonies were identified based on growth characteristics on MSA plates (golden yellow colonies), Gram stain and positive results for coagulase tube test and catalase test. The pure isolated MRSA were subjected to antibiotic susceptibility tests and biofilm formation assays.Results From our study the overall prevalence of CA-MRSA was 61.5%. Higher frequency of multi-drug resistant MRSA was isolated. The biofilm producing CA-MRSA were 51.2% which showed high drug resistance and rest (48.7%) were non-biofilm producers. There was significant association in biofilm production with multi-drug resistance (p&lt;0.05). Ciprofloxacin was most sensitive drug against the isolates which was statistically significant (p&lt;0.05). The resistant pattern of biofilm producers reported high ability of multi-drug resistance compared to non-biofilm producers (p&lt;0.05). Microtitre plate method was found to be gold standard over tube and congo red agar method for screening biofilm formation. Surprisingly the emergence of VISA and VRSA strains were significantly reported from our study. The prevalence of VISA and VRSA among CA-MRSA was found to be 49.5% and 40.6% respectively among the isolates which indicates the failure of Vancomycin drug in clinical therapy.Conclusions The prevalence of CA-MRSA was found more in barbers followed by beauticians and municipal waste workers in comparison to healthy controls. This study reported the higher carriage of CA-MRSA in potential risk population along with emergence of VISA and VRSA strains. Improvement in personal hygiene and formulation of appropriate health policy helps to prevent CA-MRSA infection. This study concludes that CA-MRSA is still emerging with multi-drug resistance.</jats:p

Surfactant Production by &lt;em&gt;Bacillus subtilis&lt;/em&gt; Using Very Low Grade and Cheap Substrate

Bio-surfactants are surface-active molecules which are produced by the wide range of microbes including bacteria, fungi, and yeast. This study was conducted to identify bio-surfactants by Bacillus subtilis combined with use of cheap substrates and industrial wastes (Mustard cake, Whey and Soya cake) which are found locally in Nepal. Bacillus subtilis, one of the most potential bio-surfactants producer; was isolated from soil sample of hydrocarbon contaminated site. Isolates were grown in a Minimal Salt Media (MSM) with 10% (v/v) mustard oil cake, whey and soya cake separately. The presence and potential of surfactant was determined by the oil spreading technique, emulsification index (%E24) and surface tension measurement. It was revealed that the surface tensions of cell free extract were 54.41, 60.02 and 56.64 mN/m for from mustard cake, whey and soya cake respectively as compared to distilled water (72.09) at 25oC. The emulsification index values are was found to be highest in engine oil from the bio-surfactant extracted from mustard cake, soya cake and whey respectively. Similarly, mustard oil showed the lowest value of emulsification index. The highest emulsification activity was shown in mustard oil i.e. 1.13 from the cell free extract from mustard oil and lowest in engine oil i.e., 0.07, by the extract from soya cake medium, when measured in spectrophotometer at 540 nm. In conclusion, strain of Bacillus subtilis was found to be the potential surface active agent producers on the mustard oil cake, which can be useful medium for various environmental, food and industrial processes.</jats:p