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ThesisItem Restricted Use of bioinsecticides in compost for button mushroom cultivation(Punjab Agricultural University, Ludhiana, 2020) Simran; Sharma, ShivaniAgaricus bisporus is the most commonly cultivated edible variety in the world which possesses an ability to grow on agricultural wastes. Therefore, the present study has dealt with the use of pre botanical insecticides such as neem powder, vinegar, castor powder and two biological insecticides such as Bacillus thuringiensis and Beauveria bassiana to study its effect on mycelial growth of A. bisporus on PDA medium and wheat straw based compost. Maximum growth of 55mm and 29mm was recorded with neem powder on PDA and compost respectively. However, vinegar and Bacillus thuringiensis also showed significant mycelial run on PDA as well as compost. The natural insecticides were supplemented @0.5 to 4.5% (w/w) in ready compost and spawned @1.0% in compost. The supplementation of neem powder @1.5% showed significantly improved yield potential 43.40% and increased number of fruit bodies 37.55%. In case of castor powder (0.5 to 4.5% v/w), supplementation @ 4.5% (v/w) showed an increase of 23.22% in yield of button mushroom. An increase upto 20.23% and 46.42% in yield and fruit bodies was observed when vinegar was supplemented @2.5%. Bacillus thuringiensis supplementation @2.5% showed maximum yield potential 33.96%. Similarly, Beauveria bassiana was supplemented at a population of 3.4×107 cfu/ml @0.54.5% (v/w) in compost which significantly improved the yield upto 14.44% and 12.62% when Beauveria bassiana was supplemented @1.5% and 0.5% at the time of spawning. Thus, it was concluded from our study that plant-derived products can be recommended to substitute for synthetic chemicals in the commercial production of edible mushrooms.ThesisItem Open Access Evaluation of antioxidant activity in selenium biofortified oyster mushrooms; Pleurotus spp.(Punjab Agricultural University, Ludhiana, 2020) Sravani, Anduri; Sharma, ShivaniPleurotus florida, P. eryngii, P. cornucopiae and P. djamor were studied for comparing the effect of selenium biofortification on antioxidant activity. Maximum radial growth, biomass production and total soluble protein content of mycelium were observed @ 5 mg L-1 in P. florida (9.00 mm/day, 88.0 g L-1 and 2.28 mg g-1 respectively), @ 10 mg L-1 in P. cornucopiae (8.02 mm/day, 58.3g L-1 and 1.94 mg g-1 respectively), P. djamor (7.11 mm/day, 88.5g L-1and 1.97 mg g-1 respectively) and @ 20 mg L-1 sodium selenate supplementation in P. eryngii (8.56 mm/day, 77.0 g L-1 and 2.55 mg g-1 respectively). The highest Se content was observed at 20 mg L-1 of Se concentration in P. djamor (920.32 µg g-1dw), followed by P. florida (880.61 µg g-1dw), P. eryngii (850.27 µg g-1dw) and P. cornucopiae (792.72 µg g-1dw). SEM studies showed that 10 mg L-1 Se concentration was best for the growth of P. florida, P. eryngii, P. djamor and P. cornucopiae and the mycelial texture became coarser and fragile in response to increased concentration of Se. EDS analysis showed that the % weight and atomic % of Se were highest in 10 mg L-1 Se supplementation treatment in all the four Pleurotus spp. There was no significant difference in the biological efficiency (%) of P. florida (66.50 vs 64.57), P. cornucopiae (36.74 vs 33.11), P. eryngii (30.41 vs 27.16) and P. djamor (30.14 vs 29.16) cultivated on Se-enriched and non-enriched wheat straw respectively. The total soluble protein content, total phenolic content, flavonoid content and free radical scavenging activity (%) were found to be highest in Se enriched fruit bodies when compared to control samples in all the four Pleurotus spp. Elemental analysis of Se biofortified Pleurotus mushrooms using SEM-EDS showed signals characteristic for selenium on the surface of P. florida and P. djamor conforming that the selenium was incorporated into the cell wall of these mushroom fruiting bodies and were absent in P. eryngii and P. cornucopiae. The Se content was found to be 22.34 µg g-1dw in Se enriched wheat straw and 0.059 µg g-1dw in respective non-enriched wheat straw. Se content of Se-enriched fruit bodies were found to be higher as compared to non-enriched fruit bodies of P. florida (124.00 vs 2.48 µg g-1dw), P. eryngii (57.25 vs 0.49 µg g-1dw), P. cornucopiae (46.25 vs 0.161 µg g-1dw) and P. djamor (156.6 vs 4.33 µg g-1dw). FT-IR spectra of proteins from all the four Pleurotus spp. indicated an increase in flexibility, unfolding and hydrophilicity upon Se supplementation. Therefore, Pleurotus spp. can be used as an excellent nutraceutical with numerous health benefits.ThesisItem Open Access Use of bioinsecticides in compost for button mushroom cultivation(Punjab Agricultural University, Ludhiana, 2020) Simran; Sharma, ShivaniAgaricus bisporus is the most commonly cultivated edible variety in the world which possesses an ability to grow on agricultural wastes. Therefore, the present study has dealt with the use of pre botanical insecticides such as neem powder, vinegar, castor powder and two biological insecticides such as Bacillus thuringiensis and Beauveria bassiana to study its effect on mycelial growth of A. bisporus on PDA medium and wheat straw based compost. Maximum growth of 55mm and 29mm was recorded with neem powder on PDA and compost respectively. However, vinegar and Bacillus thuringiensis also showed significant mycelial run on PDA as well as compost. The natural insecticides were supplemented @0.5 to 4.5% (w/w) in ready compost and spawned @1.0% in compost. The supplementation of neem powder @1.5% showed significantly improved yield potential 43.40% and increased number of fruit bodies 37.55%. In case of castor powder (0.5 to 4.5% v/w), supplementation @ 4.5% (v/w) showed an increase of 23.22% in yield of button mushroom. An increase upto 20.23% and 46.42% in yield and fruit bodies was observed when vinegar was supplemented @2.5%. Bacillus thuringiensis supplementation @2.5% showed maximum yield potential 33.96%. Similarly, Beauveria bassiana was supplemented at a population of 3.4×107 cfu/ml @0.5- 4.5% (v/w) in compost which significantly improved the yield upto 14.44% and 12.62% when Beauveria bassiana was supplemented @1.5% and 0.5% at the time of spawning. Thus, it was concluded from our study that plant-derived products can be recommended to substitute for synthetic chemicals in the commercial production of edible mushrooms.ThesisItem Restricted Upscaling and comparison of beer production by Saccharomyces cerevisiae fermentation of sweet sorghum(Punjab Agricultural University, Ludhiana, 2020) Jasmine Kaur; KeshaniBeer is the third most popular beverage in the world. It is generally made from malted barley. Tropical countries need to import barley from temperate countries for brewing which makes it an expensive process. So, brewers are searching for alternate substrates for beer production to meet the increasing demand of beer and to reduce its cost. Sweet sorghum is one of the potential substrate for beer production. In the present research, fifteen varieties of sweet sorghum grains viz. SPV 2402, SPV 2530, SPV 2596, SPV 2597, SPV 2599, SPV 2600, SPV 2601, SPV 2603, SPV 2606, SPV 2610, SPV 2611, CSV 19 SS, CSV 24SS, CSH 22 SS and SSV 84 were evaluated for their potential as a substrate for beer production. Beer was produced using sweet sorghum and pilsner malt blend in the ratio of 40:60 respectively. Out of all the sweet sorghum varieties assessed, beer made from CSV 24SS using Saccharomyces cerevisiae GP4 had highest alcohol content of 5.8% (v/v) and the highest sensorial acceptance. The CSV 24SS variety was then used for upscaled production (20 litres) of beer using S. cerevisiae GP4 fermentation. The beer produced was pale gold in color and had a pH of 4.2 and titrable acidity of 0.36%. The beer was evaluated by a panel of judges for its sensorial profile and received a mean score of 31.5 out of 36 on the Hedonic scale. The beer was subjected for shelf life studies. Few bottles of beer were force carbonated and others were kept flat and stored at 4°C. The flat beer showed greater stability during storage as compared to carbonated beer.ThesisItem Open Access Seasonal emergence of pathogenic bacteria in drinking water(Punjab Agricultural University, Ludhiana, 2020) Pandey, Swati; Sahota, Param PalThe impact of seasonal variations on microbial diversity of drinking water poses an imminent threat to public health. As such the seasonal dynamics of drinking water quality characteristics were investigated during the year 2019-2020 in Punjab. A total of N=600 drinking water samples from various sources and utilities were analysed for bacteriological and physicochemical attributes; viz heterotrophic plate counts (HPC), total coliforms (TC), faecal coliform E. coli, emerging pathogens, pH, conductivity (EC), and TDS. A total of 66.3% (n=398) samples were tested positive for the presence of coliforms. The mean HPC and TCC were found to differ significantly during seasons (summer, monsoon, and winter) of the year (P < 0.05). There were considerable seasonal variations in prevalence of indicator E. coli (16.7-60%) as well as the emerging pathogens in contaminated drinking water samples. Statistical analysis of the dataset showed significant positive correlation trends between temperature and TCC (r = 0.785), E. coli (r = 0.690) and HPC (r = 0.678). The prevalence of emerging pathogens in contaminated samples and temperature showed a significant positive correlation for Enterococcus spp., Campylobacter spp., Vibrio spp. and inverse significant correlation for Aeromonas spp., Yersinia spp., Pseudomonas spp. and Listeria spp. The bacterial pathogens Klebsiella spp., Shigella spp., Staphylococcus spp., Salmonella spp. were detected from contaminated samples throughout the year in low percentages regardless of temperature influences. The concurrence of faecal indicator E. coli and emerging bacterial pathogens in contaminated samples was weak and variable. The bacterial isolates from drinking water tested for antibiotic sensitivity assay showed MAR index range of 0.12-0.87. The commercially available UV filter was explored as a remedial technology for bacteriologically contaminated water. Its effectiveness under applicable conditions debunks the common belief about the advanced disinfection potential. The presence of injured coliforms and bacterial pathogens showing marked phenotypic and antibiotic sensitivity changes post-treatment raises the alarming concern of consumer safety. Lastly collection of a large dataset at high frequency over a period of one calendar year has enabled the characterization of previously undocumented seasonal dynamics in the drinking water characteristics and highlights the need for reframed standard methods to enhance pathogenspecific assessment for better risk management.ThesisItem Restricted Nano-zinc oxide (ZnO)-polymer seed coatings and their effect on soil microbes in maize (Zea mays L.)(Punjab Agricultural University, Ludhiana, 2020) Tondey, Manisha; Kalia, AnuThe present work aimed for the synthesis of ZnONPs from zinc acetate or zinc chloride as precursor salts by using various reducing agents such as natural polymers (starch, cellulose, potato-extract), bovine serum albumin, and thiourea. On UV-Vis absorption spectroscopy analysis, the prepared ZnONPs exhibited characteristic absorption peaks ranging from 210 to 360 nm wavelength. The FT-IR spectroscopy of the synthesized ZnONPs depicted presence of Zn metal-oxide functional groups. Transmission electron microscopy analysis revealed quasispherical shapes of ZnONPs with an average particle size of <100 nm. The synthesized ZnONPs were also evaluated for their antimicrobial potential for plant beneficial and phyto-pathogenic bacterial and fungal cultures. Wet-chemistry derived ZnONPs (at 40 mg L-1) exhibited antimicrobial activity against Dickeya sp. while at 20 mg L-1 concentration radial mycelial growth of Macrophomina phaseolina was inhibited. Apart from antimicrobial potential, the free radical scavenging or antioxidant activity of prepared ZnONPs as determined through DPPH radical scavenging method. It was observed to be vary considerably among the ZnONPs synthesized using different reducing agents and hence is anticipated to positively affect seed germination. The effect of application of ZnONPs as seed priming and coating treatment on germination and seedling growth traits was evaluated for two maize varieties PMH-1 and J-1006 using different concentrations (0, 5, 10, 20, 40 mg L-1) of ZnO NPs and bulk ZnSO4 suspensions under laboratory conditions. Seed coating with 20 mg L-1 of ZnONPs exhibited significantly higher percent germination, root and shoot length, and fresh and dry weight of root besides number of lateral roots of the maize seedlings (variety J-1006) as compared to bulk ZnSO4 treatments. Further, ZnONPs priming and coating treatments were also evaluated for enhancing vegetative growth and yield attributing parameters under field conditions during spring and kharif seasons in the year of 2017-18 and 2018-19. The application of ZnONPs (at 40 mg L-1 concentration) significantly enhanced the total chlorophyll content, available soil nutrient content (Nitrogen and Phosphorus), improved the total soil microbial counts and soil enzyme activities(dehydrogenase, and acid and alkaline phosphatase enzyme activities). Whereas, a significant increase in the available soil potassium, and zinc contents and numerical increase in vegetative and yield parameters (number of plants, plant height, stover yield, plant biomass) was recorded on treatment of seeds with ZnONPs at 20 mg L-1 in maize variety J-1006 as compared to bulk ZnSO4 and control treatments. These findings suggest positive effect on growth and yield attributing characters in maize on seed priming and coating with ZnONPs. Therefore, use of ZnONPs at low concentrations as seed priming/ coating agent can effectively improve productivity and soil health.ThesisItem Open Access Ester Profile of indigenous Saccharomyces cerevisiae strains isolated from traditional alcoholic beverages(Punjab Agricultural University, Ludhiana, 2020) Harmanpreet Kaur; KeshaniRed wine was prepared from the red grapes var. Zinfandel by the action of twenty-three yeast cultures isolated from traditional alcoholic beverages from Lahaul & Spiti and Bada Bhangal region. The yeasts were identified on the basis of morphological studies with respect to colony characteristics, characteristics in broth culture, and cell shape as well as biochemical characteristics. Extracted juice was subjected to skin treatment for 16h to improve the color intensity of juice. The later was fermented by twenty-three different Saccharomyces cerevisiae strains under optimized conditions viz. 5% (v/v) inoculum size, 50 mg/100 ml DAHP was supplemented at 28 ˚C. The prepared wine was compared for their alcoholic content as well as their overall acceptance on the basis of appearance, mouthfeel and flavor by sensory evaluation. The ester analysis of prepared wine was done by GC-MS technique. Esters such as ethyl acetate, isoamyl acetate, phenyl ethyl acetate, ethyl caprylate and ethyl caproate as well as the other volatile compounds were reported out of which ethyl acetate and isoamyl acetate were predominant. Maximum alcohol content (11.03% v/v) and fermentation efficiency (86.16%) was noticed in the wine produced by strain code AB. It also produces enough esters viz. ethyl acetate (36.55 mg/l) and isoamyl acetate (940 mg/l) to provide suitable flavor to the wine which leads to score maximum points (7) on the “Hedonic-scale” and was considered best among all.ThesisItem Open Access Bioprocessing of Indian gooseberry for the production of lactic acid beverage and pickles(Punjab Agricultural University, Ludhiana, 2019) Kirandeep Kaur; Sahota, Param PalThe plant-based fermented beverages have received considerable interest for their rich nutrition, high content of soluble and insoluble fibers and for their fermentability by lactic acid bacteria (LAB). A set of ten allochthonous lactic acid bacterial strains phenotypically and genotypically characterized for their probiotic potential were used as starter culture for the optimization and production of Indian gooseberry-based functional beverage and lacto-pickle. The optimized bioprocess comprised of gooseberry juice blend [gooseberry juice 2: ginger extract 1: guava juice 2], dilution ratio (1:3), condiment concentration (0.7%), pasteurized at 82ºC for 10 seconds and 5.0% (v/v) (106 CFU/mL) active starter culture for fermentation at 37ºC for 24 hr. The microbiological and physicochemical parameters of stored beverage (90 days) were; 1ºBrix, Titrable acidity 0.30-0.89 %, pH 6.0 -2.6, Total Sugars 44.3-35.3 mg/100mL, Reducing sugars 5.4-10.7 mg/100mL and antioxidant activity (85-67%), Total polyphenolic content (42.5-32.6 mg GAE/100mL),Total flavanoids (14.5-28.7 mg/100mL) with LAB capable of maintaining cell counts of 7.39 log CFU/mL. The bioprocess optimized for the Indian gooseberry lacto pickle as 5.0% (v/v) (106 CFU/mL) active starter culture and 5% brine solution for fermentation at 37ºC for 24 hr. The microbiological and physicochemical parameters of stored lacto-pickle (90 days) were; LAB count-6.22-7.27 log CFU/mL, 1ºBrix, Titrable acidity 0.24-0.64 %, pH 7.0-2.8, Total Sugars 20.1-11.6 mg/100g, Reducing sugars 2.60-14.6 mg/100g, viz. nutraceuticals Total antioxidant activity 86.1- 90.2%, Total polyphenolic content 16.6-37.8 mg GAE/100g and Total flavanoids 4.007-55.7 mg/100g. The overall acceptability of beverage and pickle was 7±0.2 on nine point hedonic scale. The bio-interventions: Gooseberry beverage and pickle endowed with organic acids, metabolites, antioxidants, polyphenols and flavonoids showed antimicrobial activity against food borne pathogens Staphylococcus aureus MTCC3906, Listeria monocytogenes MTCC657, Klebsiella pneumoniae MTCC109, Escherichia coli MTCC443, Aeromonas hydrophila MTCC173 as well as arrested the initial phase of Molt-4 cancer cell lines, down regulating the expression of proto-oncogenes and up regulating the tumor suppressor gene exhibiting the antitumorigenic effect.ThesisItem Restricted Bio utilization of honey processing waste into mead(Punjab Agricultural University, Ludhiana, 2020) Dhillon, Sunjit Kaur; Kocher, G.S.The honey which goes waste during processing as well as storage was selected as the substrate for the present study on the basis of evaluation of physico-chemical parameters such as total soluble solids, total sugars, reducing sugars, moisture content, refractive index, acidity, pH, specific gravity, nitrogen content, phosphorus content, phenols, antioxidant activity, total yeast count and total microbial count. The study of yeast growth dynamics on the basis of absorbance measurements, haemocytometric count and consumption of reducing sugars led to the selection of Saccharomyces cerevisiae KY069279 as the best suited strain for growth on honey in comparison to other three yeast strains viz. S. cerevisiae MTCC 11815, D7 and Meyerozyma caribbica PY1. The yeast strain S. cerevisiae KY069279 showed the maximum growth rate (0.16 log cells/h), sugar consumption rate (0.078 g/h) and yield (0.054 log cells/h/g sugar consumed) on honey. Effect of addition of various supplements like grape juice, apple juice, yeast nitrogen base (YNB) and resins was studied whereby, addition of yeast nitrogen base led to the completion of fermentation in 8 days. Hence, YNB was chosen as the preferred nitrogen source for fermentation on honey. Optimization of mead fermentation parameters like initial brix, inoculum size, YNB concentration and acidity was carried out using Response Surface Methodology. The model revealed that an initial sugar level of 15°B, inoculum size 10%, YNB concentration 20.03 % and acidity 0.2% were statistically the optimum conditions for honey fermentation scale up experiments with a desirability of 95.7%. An ethanol content of 8.79% (v/v) with an ethanol yield 0.486 g/g and fermentation efficiency of 95.23 % was observed during the scale up experiments carried out at 3L scale. The prepared mead was evaluated for the physico-chemical parameters over a period of 90 days at regular intervals of one month. Shelf life studies revealed that the mead retained its quality over a tested period of three months. Sensory analysis by a panel of 6 panelists categorized mead at 0 day and mead at 90 days as ―moderately liked‖ and ―liked very much‖ with mean scores of 7.31 and 8.01, respectively on the 10 point Hedonic scale suggesting that mead sensory quality increased during storage. Highest antioxidant activity in terms of AEAC value was observed in case of packaged honey (33%), followed by waste honey (28%) and mead (22%) w.r.t ascorbic acid (100 % v/v). Organoleptic evaluation of dehydrated samples by gas chromatography mass spectroscopy listed esters, alcohols, volatile fatty acids, acetates, amines, amides, terpenes and lactones as the major constituents of mead.