A sustainable approach of turning potato waste towards bioethanol production using indigenous microbes of Himachal Pradesh, India.

Potato peel waste is one of the zero-value wastes with the potential of bioethanol production through the Waste to Energy (WtE) approach. The newly isolated, phenotypically characterized, and molecular identified high-altitude strain, B. amyloliquefaciens, shown promising starch hydrolysis (12.06 g/L reducing sugars) over acid hydrolysis and is capable of working at 30-50 °C and pH 6.0-8.0. The ethanol production by Acinetobacter sp. (a newly isolated, phenotypically characterized, molecular identified) has been modelled and optimized through the central composite design of response surface methodology by taking the fermentation variables as input variables and ethanol yield as the output variable. The ethanol production by Acinetobacter sp. showcased a non-linear relationship of fermentation variables with the ethanol yield (5.83 g/L) with a 99.11% desirability function (R2) and 97.50 adj. R2 values. Optimal fermentation variables of 38.8% substrate concentration, 7% inoculum, pH 5.45 have been utilized for bioethanol production in 55.27 h at 27 °C. Overall, the present study evaluated the efficiency of newly isolated, indigenous extremophilic microbes of The Himalayan region in sustainable bioethanol production from zero-value waste "Potato peel waste" through the WtE approach. Moreover, the present study introduces the promising, unexplored extremophilic microbial strains with the starch-hydrolyzing and fermentation capabilities to bioethanol biorefinery.

Compliance assessment of cigarette and other tobacco products act in public places of Alwar district of Rajasthan.

BACKGROUND: The Government of India has taken various initiatives for tobacco control by enacting comprehensive tobacco control legislation (Cigarette and Other Tobacco Products Act [COTPA], 2003). OBJECTIVES: The aim of this study was to assess the level of compliance of Sections 4, 5, 6-a, and 6-b, and 7, 8, and 9 of COTPA with respect to public places, educational institutes, point of sale (PoS), and warning on packaging (COTPA) in public places of Alwar District of Rajasthan. METHODS: A cross-sectional observational study was conducted in 2014 in Alwar city and four blocks of the district. The study was done around 365 public places for observing the compliance of Section 4 of COTPA, 357 educational institutions for observing the compliance of Section 6-b of COTPA, and 357 tobacco retailers for observing the compliance of Sections 5 and 6-a of COTPA. RESULTS: The criteria for the evaluation (the core indicators) and decision criteria for a district to qualify for the "Smoke free" status include six parameters. From the total of 365 places visited, 90% places displayed the "No-smoking" signage and out of total 328 places, 99% were as per the COTPA specification. Alwar city, Ramgarh, Thanagaji, and Alwar rural block followed the compliance of Section 4. The PoS visited Alwar district displayed 93% (332) signage and all the displayed signage followed the COTPA compliance. In Alwar city, Thanagaji, Ramgarh, and Alwar rural block, the compliance of Section 6-a was above 90%. The compliance of Section 6-b was above 90% in Alwar city, Ramgarh, Thanagaji, and Alwar rural block. Ninety-three percent (332) of the PoS did not display tobacco advertisement in Alwar district, which is a positive sign of COTPA compliance. CONCLUSION: This finding suggest a high level of compliance of Section 4, Section 5, Section 6-a, and Section 6-b of COTPA at Alwar district.

Genetic diversity and bottleneck studies in endangered Bhutia and Manipuri pony breeds.

For studying the genetic diversity and bottleneck problem in Bhutia and Manipuri pony breeds of India, we analysed DNA samples of 34 Bhutia and 50 Manipuri, true to breed, ponies using 47 polymorphic microsatellite markers. All the microsatellites were observed to be highly polymorphic in nature in both Bhutia and Manipuri breeds with mean no. of alleles as 8.702 ± 0.0493 and 8.416 ± 0.0548 respectively. Genetic diversity values in terms of heterozygosity values within individual breeds were also high with very low inbreeding (Fis 0.102 and 0.055 in Bhutia and Manipuri ponies, respectively). Number of alleles in both the populations together ranged from 3 to 18 with an average of 10.851 ± 1.583 per locus. The mean effective number of alleles was observed to 5.34 ± 0.253. All loci except ASB017 and HTG004 showed high values of allele richness (>5.0). The mean observed and expected heterozygosities were 0.7159 ± 0.022, 0.7986 ± 0.011 (Levene's) and 0.7936 ± 0.011 (Nei's), respectively. The high mean values of heterozygosity indicated the presence of high genetic diversity in both the pony populations. The overall mean value of within-population inbreeding estimates (Fis) was low (0.101 ± 0.023) indicating low to moderate level of inbreeding. Bottleneck studies revealed that no recent bottleneck problem has taken place in both the populations. Both pony populations were found to be in mutation drift equilibrium. The study reveals that both the pony breeds have high diversity and timely action needs to be taken to conserve them.

Evaluation of a new lipase from Staphylococcus sp. for detergent additive capability.

Lipases are the enzymes of choice for laundry detergent industries owing to their triglyceride removing ability from the soiled fabric which eventually reduces the usage of phosphate-based chemical cleansers in the detergent formulation. In the present study, a partially purified bacterial lipase from Staphylococcus arlettae JPBW-1 isolated from the rock salt mine has been assessed for its triglyceride removing ability by developing a presoak solution so as to use lipase as an additive in laundry detergent formulations. The effects of selected surfactants, commercial detergents, and oxidizing agents on lipase stability were studied in a preliminary evaluation for its further usage in the industrial environment. Partially purified lipase has shown good stability in presence of surfactants, commercial detergents, and oxidizing agents. Washing efficiency has been found to be enhanced while using lipase with 0.5% nonionic detergent than the anioinic detergent. The wash performance using 0.5% wheel with 40 U lipase at 40°C in 45 min results in maximum oil removal (62%) from the soiled cotton fabric. Hence, the present study opens the new era in enzyme-based detergent sector for formulation of chemical-free detergent using alkaline bacterial lipase.

Production and characterization of a halo-, solvent-, thermo-tolerant alkaline lipase by Staphylococcus arlettae JPBW-1, isolated from rock salt mine.

Studies on lipase production and characterization were carried out with a bacterial strain Staphylococcus arlettae JPBW-1 isolated from rock salt mine, Darang, HP, India. Higher lipase activity has been obtained using 10 % inoculum with 5 % of soybean oil as carbon source utilizing a pH 8.0 in 3 h at 35 °C and 100 rpm through submerged fermentation. Partially purified S. arlettae lipase has been found to be active over a broad range of temperature (30-90 °C), pH (7.0-12.0) and NaCl concentration (0-20 %). It has shown extreme stability with solvents such as benzene, xylene, n-hexane, methanol, ethanol and toluene up to 30 % (v/v). The lipase activity has been found to be inhibited by metal ions of K(+), Co(2+) and Fe (2+) and stimulated by Mn(2+), Ca(2+) and Hg(2+). Lipase activity has been diminished with denaturants, but enhanced effect has been observed with surfactants, such as Tween 80, Tween 40 and chelator EDTA. The K m and V max values were found to be 7.05 mM and 2.67 mmol/min, respectively. Thus, the lipase from S. arlettae may have considerable potential for industrial application from the perspectives of its tolerance towards industrial extreme conditions of pH, temperature, salt and solvent.

Modelling and Optimization Studies on a Novel Lipase Production by Staphylococcus arlettae through Submerged Fermentation.

Microbial enzymes from extremophilic regions such as hot spring serve as an important source of various stable and valuable industrial enzymes. The present paper encompasses the modeling and optimization approach for production of halophilic, solvent, tolerant, and alkaline lipase from Staphylococcus arlettae through response surface methodology integrated nature inspired genetic algorithm. Response surface model based on central composite design has been developed by considering the individual and interaction effects of fermentation conditions on lipase production through submerged fermentation. The validated input space of response surface model (with R (2) value of 96.6%) has been utilized for optimization through genetic algorithm. An optimum lipase yield of 6.5 U/mL has been obtained using binary coded genetic algorithm predicted conditions of 9.39% inoculum with the oil concentration of 10.285% in 2.99 hrs using pH of 7.32 at 38.8°C. This outcome could contribute to introducing this extremophilic lipase (halophilic, solvent, and tolerant) to industrial biotechnology sector and will be a probable choice for different food, detergent, chemical, and pharmaceutical industries. The present work also demonstrated the feasibility of statistical design tools integration with computational tools for optimization of fermentation conditions for maximum lipase production.

A simplified chair-side remount technique using customized mounting platforms.

Correct occlusal relationships are part of the successful prosthetic treatment for edentulous patients. Fabrication of complete dentures comprises of clinical and laboratory procedures that should be executed accurately for achieving success with fabricated dentures. Errors occurring during the clinical and laboratory procedures of a denture may subsequently lead to the occlusal errors in the final prosthesis. These occlusal errors can be corrected in two ways: i) in patient's mouth ii) by recording new centric relation and remounting dentures on an articulator. The latter method is more feasible because the mobility of denture base on the mucosa in oral cavity does not permit the identification of premature contacts in centric occlusion and tooth guided eccentric excursions. This article describes a modest and effective clinical chair-side remount procedure using customized mounting platforms.

Microsatellite markers based genetic diversity and bottleneck studies in Zanskari pony.

Genetic diversity in Zanskari pony breed was evaluated at 48 microsatellite loci using fifty adult, healthy and unrelated animals. Allele frequency data was used to detect genetic diversity and bottleneck. The estimated average number of alleles (±s.e.) was 8.5208±2.5010 with a total of 409 alleles. A high level of genetic diversity within this breed was observed in terms of number of alleles, observed heterozygosity (0.6763±0.1704), expected Leven's heterozygosity (0.7724±0.795), expected Nei's heterozygosity (0.7644±0.0787) and polymorphism information content (>0.5). In-breeding coefficient (F(is)) was 0.115±0.0209, suggesting moderately high in-breeding in Zanskari breed. Although analysis of bottleneck revealed no bottleneck in recent past but population of Zanskari ponies has decreased drastically and only a few thousand pure-bred animals are left. The information is useful for proposing effective population management strategies for future.

Outbreak of Abortions and Infertility in Thoroughbred Mares Associated with Waterborne Aeromonas hydrophila.

At a thoroughbred equine breeding farm near Hissar (Haryana), three mares aborted in their seventh month of pregnancy. The vaginal swabs of all aborted mares, and stomach contents, heart blood, liver, spleen and placenta of aborted fetuses yielded pure culture of Aeromonas hydrophila. In addition, A. hydrophila was also isolated from the vaginal swabs of three repeat breeding mares and faecal sample of a diarrheic foal. The source of infection was possibly water supply as all the water samples collected from taps, mother tank and storage tank were found to be positive for A. hydrophila. The antibiogram of all the isolates was similar showing resistance to ampicillin, carbenicillin, gentamicin, kanamycin and amikacin but sensitive to chloramphenicol, ciprofloxacin, cefuroxime, ceftriaxone, cotrimoxazole, cotrimazine, nitrofurantoin, streptomycin and tetracycline. All the 20 sera samples collected from three aborted and three repeat breeding, and eight in-contact mares, a diarrheic foal, three cows and two male buffaloes maintained at the same farm contained antibodies against A. hydrophila with titres ranging from 80 to 640. The water supply was instantly chlorinated using 0.05% sodium hypochlorite for three consecutive days and all the culturally positive mares were treated with intravaginal administration of 1 g ciprofloxacin, while the foal was given nitrofurantoin for three days. After one month, A. hydrophila could not be isolated either from mares or from their environment and antibody titre in all the seropositive animals showed a declining trend. Later, all the aborted and repeat breeding mares were confirmed to be pregnant. Thus, the present study indicated that water-borne A. hydrophila might be associated with equine abortions and infertility, and diarrhea in newborn foals.

Genetic diversity and population structure of three Indian horse breeds.

The genetic relationships of three Indian horse breeds-Marwari, Spiti, and Kathiawari were studied by genotyping 96 individuals with 20 polymorphic microsatellite markers. A total of 157 alleles were detected across 20 polymorphic loci. The Marwari population showed the highest allelic diversity (A = 5.7 and Ar = 5.14), followed by Spiti (A = 4.9 and Ar = 4.74) and Kathiawari (A = 4.1 and Ar = 3.82). The gene diversity was highest in the Spiti population (He = 0.67), followed by Marwari (He = 0.66) and Kathiawari (He = 0.59). Within population inbreeding estimates (f) in Marwari, Spiti and Kathiawari breeds were 0.18, 0.08, and 0.07, respectively, suggesting high level of inbreeding in these breeds. Analysis of bottleneck revealed evidence of recent bottleneck in Spiti and Kathiawari populations. Pair-wise Fst analysis, AMOVA and assignment tests demonstrated high genetic differentiation and low gene flow between populations. The information about genetic diversity and population structure will be useful for the future development of effective breeding management in order to preserve these Indian horse breeds.