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ThesisItem Open Access Extraction and Characterization of Essential Oil and Phytochemicals From Galgal (Citrus pseudolimon) Peel and Seeds for Utilization in Food Formulations(Punjab Agricultural University, 2024) Grover, Sumit; Dr Poonam A. SachdevThe study investigated the extraction and characterization of essential oil and phytochemicals from galgal (Citrus pseudolimon) peel and seeds for potential utilization in food formulations. The focus was on identifying the most promising cultivar, evaluating antioxidant activities, mineral content, volatile components, phytochemical extraction, fatty acid composition, sterol profiling, and various extraction methods for essential oil and microencapsulation, thereof. Among the studied galgal cultivars (HRG, PBG & HPG), the one from Punjab (PBG) exhibited superior properties in terms of phytochemicals, essential oil, and bioactive compounds. The antioxidant properties of galgal fruits were evaluated using different methods, including DPPH, FRAP, ABTS, and reducing power assays. PBG (whole fruit) demonstrated the highest antioxidant activity. PBG cultivar showing the highest levels of potassium, sodium, and calcium content analyzed by ICP-OES. Gas Chromatography-Mass Spectrometry (GCMS) analysis of galgal juice revealed the presence of various volatile compounds. PBG was rich in caryophyllene, D-limonene, á-Pinene, and other minor components. Different solvents i.e hexane, methanol, and DMSO, were used to extract phytochemicals from galgal peel. PBG primarily contained D-limonene, caryophyllene, and á-Myrcene. Fatty acid composition in the seed oil of different galgal cultivars was found to be similar, with SFA:MUFA:PUFA ratios meeting WHO/ICMR recommendations. GCMS analysis identified various sterols in the seed oil, including tetraponerine, squalene, campesterol, stigmasterol, β-sitosterol, and others. Essential oil was extracted from galgal peel using different methods, with supercritical CO2 extraction yielding the highest percentage whereas, Clevenger technique was also found to be effective and cost-efficient. GCMS analysis of essential oil revealed the presence of compounds like D-Limonene, Trans-á-Ocimene, à-Pinene, çTerpinene, Linalool, and more. Essential oil from PBG exhibited the highest antioxidant and antimicrobial activities. Microemulsions of essential oil using maltodextrin, gum Arabic, and soy lecithin were prepared and most stable emulsion was used for spray drying for the formation of microencapsulated powder. These extracted phytochemicals were added in yoghurt. The incorporation of microencapsulated powder into food products such as desi ghee, canola oil, and bread spread resulted in enhanced shelf life and sensory quality. This microencapsulated powder could serve as a natural preservative in food formulations. The research aimed to harness the potential of galgal peel and seeds for their phytochemical and essential oil content, demonstrated their utility as natural additives and preservatives in various food products. The Punjab cultivar, in particular, showed promising results across multiple parameters.ThesisItem Open Access Studies on avian diversity, biology and behaviour along Beas river conservation reserve(Punjab Agricultural University, Ludhiana, 2021) Sachin Kumar; Kler, Tejdeep KaurRiver Beas was declared as conservation reserve in year 2017 under section 36A of Wildlife Protection Act, 1972 but no study is available on the avian diversity along river. Therefore, present study was conducted from April 2019 to March 2021along Beas conservation reserve to understand the avian diversity in various sub habitats present along the river by selecting three locations i.e. Location I (Talwara: In foot hills of Shiwaliks), Location II (Beas: near urban settlement) and Location III (Harike: near rural settlement). Each location was further divided as crop fields, riparian, shallow water, edge, island and river. Study revealed that river Beas provides shelter to 245 bird species where 228 species were recorded at Location I with 40 exclusive species, 126 at Location II and 202 at Location III with 13 exclusive species. Crop fields had avian diversity of 104 at Location I, 78 at Location II and 94 at Location III. Riparian zone of river harbours 143 at location I, 83 at location II and 117 at location III. Shallow water river zone had 110 at location I, 126 at location II and 58 at location III. During this study a total of 113 species were recorded to exploiting edge sub habitat and 85 species Island sub habitat for various activities while 126 in shallow water sub habitat of riverine system which shows that the edge and island habitats are equally important from avian conservation point of view. Both Island and edge habitat are made of relatively very small area as compare to overall river habitat zone therefore these sub habitats are easy to manage but need attention during conservation planning. Crop fields and riparian habitats support more single species roosters and single group roosters. Riverine habitats (Edge, Shallow water and Islands) supports more mix group roosters. A total of 527 nests were recorded during study period having 193 nests at Talwara, 115 at Beas and 219 at Harike. Nesting of birds was niche area specific where colony nesters were tree species specific including Baya weaver nesting was specific to Date Palm and Cattle Egret nesting was specific to Kikar. Riparian zone of Talwara was wide and had minimum anthropogenic activities therefore harbour maximum of 143 species in riparian zone while Harike had wide shallow water zone therefore maximum of 126 water dependent species as compare to other two locations. Location II had almost lowest values for all diversity indexes under study (Abundance, Richness, Shannon index, Simpson index) in comparison to Location I and III shows the effect of proximity of human habitation on avian diversity. Though the all six sub-habitats understudy were adjoining to each other still a clear distinction of niche area preference was recorded during study. The birds may enter the adjoin niche but overall abundance remains similar for preferred niche for each bird species. The study concludes that significant number of species recorded have specific niche area requirements that are completed by the river’s sub-habitats including feeding, foraging, roosting and nesting therefore conservation plans should be designed in such a way that whole area come under protection as a single unit while maintaining its sub habitat identity.ThesisItem Open Access Technologies for utilization of hull less barley for high fibre foods(Punjab Agricultural University, Ludhiana, 2021) Mahamudu Mohamed Sasamalo; Amarjeet KaurBarley (Hordeum vulgare) is the fourth significant cereal cultivated for human use next to wheat, rice and maize. The demands of barley production keep on increasing day to day due to its diverse utilities, nutritive value and ever-increasing demand. Keeping these in view Punjab Agricultural University has developed two hull less barley, variety PL 891 and BHS 352. These two hull less barley varieties PL 891 and BHS 352 were procured from Department of Plant Breeding and Genetics and were given treatments viz. germination, pressure cooking, acid and alkali and milled into flour. Milled sample were analyzed for chemical composition, functional and rheological characteristics. Barley flour was stored and estimated for shelf life. The effect of treatments for changes in functional, nutritional and antinutritional was studied. All the treatments showed increase in protein content, crude fiber and dietary fiber ranging from 12.5912.77%, 4.06-4.52% and 17.48-23.8%, respectively. β-glucan content improved after treatments; ranging from 4.02-5.05%. Processing caused 33.7-74.3%, 24.4-81.4 %, 75.7-84.6% and 43.173.1% reduction in antinutritional factors such as tannins, saponins, phytic acid and trypsin inhibitor, respectively. Notably, the mineral bioavailability as well the in-vitro protein digestibility and the antioxidant potential improved with the processing. Food products such as chapatti, flat bread and cookies were prepared by 30% incorporation level. Barley drink was prepared by using 100% roasted barley grains. Among the food products prepared, for chapatti all the treatments were accepted by the panelist with the highest overall acceptability of 8.50 from those prepared from germinated flour, for flat bread and cookies those prepared from alkali treated grains had highest overall acceptability of 8.55 and 8.50, respectively. Between hull less barley varieties, variety PL 891 was found better in preparation of high fiber chapatti, flat bread and cookies and variety BHS 352 was found better for preparation of fiber barley drink.ThesisItem Open Access Characterization of pea processing waste for use in food products(Punjab Agricultural University, Ludhiana, 2021) Bhullar, Harinderjeet Kaur; Amarjeet KaurUtilization of vegetable processing industrial waste remain unexplored, but their by-products have been recognized as potential source of nutrients. Thus, the present study focused on the characterization and utilization of the pea processing waste. Pea processing waste as pea peels and small peas were blanched at 80°C for 3 minutes in 5% sodium chloride solution prior to drying (60°C for 5 hours). Blanching significantly (p<0.05) reduced ascorbic acid content (22.81%), total soluble sugars, reducing sugars, non-reducing sugars by 3.44%, 4.76%, 2.28% respectively and antinutritional factors as tannins, phytates and saponins by 35.71%, 50% and 5% respectively, while enhanced the phytochemical composition and antioxidant activities (8.18%, 10.02%, 4.81% and 24.34% increase in DPPH radical scavenging activity, ABTS radical scavenging activity, ferric reducing antioxidant power and reducing power respectively). Blanched pea peel powder contained considerable amount of all essential and non-essential amino acids, micro and macro minerals and also exhibited good functional properties. Further, protein was extracted from fresh pea peel by isoelectric precipitation method at 9 solubilization pH and 4.5 precipitation pH. Protein content of extracted pea peel protein was found to be 83.25% which exhibited variety of polypeptide subunits of molecular weight between 25 and 75 kDa. The incorporation of pea peel powder into wheat flour decreases peak viscosity, final viscosity and dough stability but increases water absorption (%) and dough development time. Thermal analysis of refined wheat flour substituted with pea peel powder revealed the increase in the onset, endset and peak temperature and also rheological attributes as storage modulus (G´) and loss modulus (G´´) increased with the increase in level of incorporation in both frequency sweep and temperature sweep test. The blanched pea peel powder was further incorporated in bakery products (cookies), extruded products (noodles), fermented products (pizza base) and traditional product (chapatti). Based on the quality and sensorial attributes, incorporation level of 10%, 7.5%, 20% and 20% was found optimum in cookies, noodles, pizza base and chapatti respectively. Cookies and noodles remain acceptable beyond 4 months of storage at room temperature (30±2°C) whereas, pizza base was acceptable for 3 days at ambient conditions (25±2°C) and for 7 days at refrigeration conditions (4±2°C). Pea peel powder incorporation increased the fiber content in all prepared products (cookies, noodles, pizza base, chapatti). This study opens an avenue to boost nutritional value of food products with additional benefit of waste management and sustainable environment by utilizing pea peels.ThesisItem Open Access Characterization and valorization of super grains(Punjab Agricultural University, Ludhiana, 2021) Kataria, Ankita; Sharma, SavitaSuper grains are considered as treasure of nutrition and taste. Present research investigation was undertaken to characterize, process and evaluate the potential of super grains (quinoa, chia and teff) as ingredients for development of functional foods. The grains were subjected to thermal processing treatments (conventional heating, roasting, microwave processing and steaming) and fermentation to achieve an optimal balance of enhanced antioxidative and functional properties, and lower antinutritional factors to improve their potentiality for utilization. The effect of the aforesaid treatments was observed on the physico-chemical, functional, antinutritional and antioxidative (bioactive compounds and antioxidant activities) characteristics. Processed super grains exhibited increased crude protein and crude fat contents with reduced carbohydrate and crude fibre levels. Thermal processing and fermentation improved the hydration properties but the former significantly affected the emulsification characteristics. Phytic acid and enzymatic inhibitors reduced during thermal processing and fermentation. Saponins and tannins increased during fermentation of super grains. All the processes improved the levels of bioactive compounds, and therefore the antioxidant activities also enhanced. Microwave processing and fermentation were more effective compared to other processing techniques for obtaining the optimal balance of improved techno-biofunctionality perceived as enhancement in the antioxidant activity and improvement in the functional properties and reduction of antinutrients. These were evaluated for their morphology, rheological properties and fourier transform infra-red (FTIR) spectra which confirmed the structural, functional and compositional changes between the raw and processed super grains. Prebiotic assessment was performed by enumerating probiotic (Lactobacillus rhamnosus), enteric (Escherichia coli) and non-probiotic (Lactococcus spp.) organisms on the super grains and comparing their growth against glucose. Teff and chia exhibited prebiotic effect whereas quinoa exhibited a negative score but it enhanced the growth of Lactococcus. Raw and processed grains and flour were used in the preparation of traditional (pinni and bhujia), confectionary (nutri-bar), extruded (pasta) and fermented (simulated product and curd) products. Cooking quality of pasta was improved upon incorporation of the super grains. Sensory evaluation revealed the optimum substitution levels of super grains for all the products which were comparably acceptable with respect to control. Antioxidant assessment and FTIR spectra of simulated product showed that dairy protein (casein) could form stable complex with super grains which would be beneficial for preparation of fermented dairy products of high nutritional and functional characteristics. Therefore, it was concluded that raw and processed super grains could be utilized as potential ingredients for preparation of specialty foods.ThesisItem Open Access Conjugate biological and non-biological N-fixation using nano-TiO2 in mungbean(Punjab Agricultural University, Ludhiana, 2021) Harleen Kaur; Kalia, AnuThe present research was proposed to probe the nitrate fixing and BNF up-regulating potential of TiO2 nanoparticles to fulfill the N requirements of mungbean plant thereby reducing the application of chemical fertilizers. One type of TiO2 nanoparticles (pure anatase) was synthesized and the other type (anatse+rutile) was purchased. Both the types of nanoparticles were immobilized in different substrates and their nitrate fixing potential was determined under ambient conditions. The TiO2 nanoparticles (anatase+rutile) adsorbed on filter paper fixed the maximum amount of nitrate (523.67 ppm NO3- m-2 h-1). From thirty isolates obtained from mungbean root nodules, two multifunctional (VR-H20 and VR-H21) along with two reference cultures were assessed for changes in growth and functional traits upon exposure to the two types of TiO2 nanoparticles. The maximum enhancement in growth, protein content, P-solubilization, IAA, siderophore and gibberellic acid production was observed for TiO2 (anatase+rutile) and the maximum exopolysaccharide production was observed for TiO2 (anatase). The mungbean rhizosphere soil was treated with TiO2 (anatase+rutile) to study the effect of nanoparticle leaching on soil properties. Low concentrations of nano-TiO2 (upto 10.0 mg L-1) enhanced the microbial population (except fungi and actinomycetes), soil enzymatic activities, chemical properties, macro and micronutrient contents except available K and Zn. The qRT-PCR study soil showed that the ammonia oxidizers were more affected as compared to nitrogen fixers upon nano-TiO2 treatment. The hydroponics experiment was designed to study the conjugative effect of TiO2 nanoparticles and rhizobia (TiO2 primed and non-primed) on mungbean. Vegetative parameters such as plant fresh and dry weight, root and shoot length, number of leaves, leaf area, and the chlorophyll content were significantly affected by various treatments. The maximum N content (3.633%) was observed in control treatment with complete PNM composition. The maximum P content (2.552%) was observed in treatment with nano-TiO2 composite assembly alone and the maximum K content (5.315%) was observed in control which was statistically at par with treatment amended with nano-TiO2 suspension (4.780%). The peroxidase and superoxide dismutase activities significantly increased with various treatments of nano-TiO2 composite assemblies and rhizobial cultures. The study revealed that the TiO2 nanoparticles could be considered an unconventional and environmentally safe method of fixation of atmospheric nitrogen to form nitrate-N.ThesisItem Open Access Development of lactic acid starter culture based fermented fruit and vegetable juices(Punjab Agricultural University, Ludhiana, 2021) Modi, Ritika; Sahota, ParampalThe lactic acid fermented fruit and vegetable beverages have been developed through controlled fermentation using ten allochthonous high lactic acid producing (0.612-1.35%), phenotypically and genotypically characterized, homo-lactic- Lactic Acid Bacterial (LAB) strains as starter culture consortium. Value-added secondary metabolite enriched Turmeric, Amla, Black carrots (Kanji), and Black pearl grapes-based functional beverages were developed with improved nutritive value (15-35%) antioxidants, (21-63%) polyphenols, (20-42%) flavonoids and (24%) carotenoids; microbial quality (1010 LAB CFU/ml) and acceptability (8±0.2) with shelf life of 90 days. The optimized bioprocess parameters using Box-Behnken Design in Response Surface Methodology with 5% (v/v) (107 CFU/ml) active starter culture for Turmeric beverage -[turmeric 2%w/v), lemon juice (5%v/v), ginger juice (1.5%v/v); dilution ratio (1:3 with sterilised water); salt (1%w/v)]; Amla beverage -[amla juice: guava juice: ginger juice (1:1:1.5 %v/v); dilution ratio (1:3); salt (0.6%w/v)]; Kanji beverage -[black carrots juice (100ml); salt and rye (1.5%w/v); dilution ratio (1:3)]; Grapes beverage -[grapes juice (100ml), lemon juice (8% v/v); dilution ratio (1:1.5); salt (1.2%w/v)], pasteurized at 82ºC for 10-15 sec and fermentation at 37ºC for 28 hrs. A generic HACCP plan determining critical control points on the line was recommended as a food safety tool during the preparation of beverages. Unstructured kinetic model so developed depicts maximum LAB growth at 8th hour and highest ∆pH and Vmax on the 28th hour and 8th hour, respectively. The increase in lactic acid production (0.35, 0.55, 0.96, 0.63 %TA), reduction in pH (4.51, 4.96, 3.76, 2.64) with enhanced functional aspects based on total polyphenols (52.30, 45.58, 41.85, 52.29 gallic acid equivalents mg/100ml) and flavonoids (44.20, 31.13, 43.91, 46.96 quercitin equivalents mg/100ml) with significantly stronger scavenging activities for the 2,2diphenyl-1- picrylhydrazyl (DPPH) radical (74.25, 86.36, 86.91, 69.70%) and ferric reducing power (87.9, 94.4, 108.66, 100.3 μM FeSO4 equivalents) was observed for Turmeric, Amla, Kanji and Grapes fermented beverages, respectively. These bio-interventions showed antimicrobial activity against food borne pathogens Staphylococcus aureus MTCC3906, Listeria monocytogenes MTCC657, Klebsiella pneumonia MTCC109, Escherichia coli MTCC443, Aeromonas hydrophila MTCC173, as well arrested the initial phase of MOLT-4 and CaCo2 cancer cells lines, down regulating the expression of proto-oncogenes and up regulating the tumor suppressor gene exhibiting the antitumorigenic effect. Further, hepatoprotective and hypoglycemic effect studies showed administration of functional lactic acid fermented turmeric and amla beverages (10 ml/kg body weight) for 6 weeks, significantly reverse or reduce the physiological, metabolic damage, and histological alterations equivalent to the hepatoprotective drug Liv52 in alcohol-induced liver damaged and hypoglycemic drug glibenclamide in streptozotocin-induced diabetic Wistar rats, respectively. Further, lyophilised freeze-dried turmeric and amla fermented powders were developed which can be consumed as ready-to-use fermented beverages by reconstitution @2% and @3.2% (with sterilized water) with retention of all properties of freshly prepared beverage prepared.ThesisItem Open Access Pilot scale production and evaluation of a food biopigment from Monascus purpureus using broken rice(Punjab Agricultural University, Ludhiana, 2021) Chaudhary, Vishu; Katyal, PriyaFood colors are an important feature that fascinates consumers; however, at present synthetic colors are being used to enhance the aesthetic value, attractiveness and flavour of food. The best alternative of synthetic colors seems to be a natural and non-toxic biopigment that has the advantages of being safe and healthy. In the present study to produce biopigment at a pilot-scale, Tray-type fermenter was found to produce optimum color value (35.05 ODU/g DMS) and yield (16.80 %) with 100 % ethanol. Biopigment extraction parameters (i.e. concentration, time and temperature) were found optimal with 60 % ethanol at 60 °C for 80 min; yielding the highest biopigment color value of 49.65 ODU/g DMS (at 500 nm) having recovery of 30.60 % DMS with tray-type fermenter. The chromatography characterization of extracted biopigment by TLC revealed that biopigment comprised of three forms i.e. red, orange, and yellow biopigments and to be free of any contaminants. Physicochemical nutrients analysis of fermented biomass has shown a decrease in carbohydrate content and pH, while increase in moisture, protein, crude fat, crude fiber and ash content. Red biopigment was stable up to 100°C, can withstand 0.6 % (w/v) of various food preservatives, but sensitive to UV/ sunlight and alkaline pH >10. Qualitative test for cardiac glycosides, hormones, alkaloids, and saponins was found to be negative but existence of different bioactive components (i.e. phenolics, flavonoids and tannins contents) was confirmed. Total phenolic content was found to increase from 3.25 - 62.70 µg GAE/g, total flavonoid content was found to increase from 2.20 - 52.04 µg QUE/g and total tannin content was found to increase from 2.59 - 30.18 µg CE/g, by increasing the concentration of biopigment from 1 µg/ml to 640 µg/ml, respectively. Similarly, iron-chelating activity of red biopigment was observed to vary from 2.44 to 67.15 % with increasing biopigment concentration. Antibacterial activity of biopigment was shown against Bacillus cereus, Staphylococcus aureus, Escherichia coli, Klebsiella pneumonia and Pseudomonas aeruginosa. Antifungal activity of biopigment has been observed against Aspergillus flavus, Fusarium oxysporum, Alternaria alternata. The DPPH antioxidant assay revealed that 640 µl of 1 mg/ml, 5 mg/ml, 20 mg/ml of extracted biopigment was found to show 53.16 %, 59.69 % and 72.73 % free radical scavenging activity. Antioxidant activity was further confirmed by ABTS and FRAP antioxidant analysis. Therapeutic potential of biopigment was observed at higher concentration (50 µg/ml) with 1.905 fold increase in the value of SOD-2. Similarly, in case of GPX, an inhibitory effect has been observed with fold change value decrease from 0.627 to 0.424. The anti-inflammatory potential of red biopigment was confirmed from increased IL-10 expression ranging from 1034.584657.89 pg/ml in LNCaP cells in comparison to untreated cells. The trypan blue, MTT and Cell viability assay revealed that red biopigment significantly reduced the viability of LNCaP cells showing its potential as anticarcinogenic agent. Moreover, it was found to be free from citrinin and contain aflatoxin within the permissible limits. The present study promises the applicability of broken rice as a potential substrate for the production of "red" biopigment with additional antimicrobial, antifungal, antioxidative, anti-inflammatory and anticancerous properties. Efforts can be made to prove the safety of product by using animal model clinical studies so that the product formulation can be designed for potential application as a biocolorant.ThesisItem Open Access Effect of agronomic manipulations on crop productivity and weed dynamics in poplar (Populus deltoides Bartr.) based agroforestry system(Punjab Agricultural University, Ludhiana, 2022) Parija, Bhadra; Navneet KaurPresent study entitled “Effect of agronomic manipulations on crop productivity and weed dynamics in poplar (Populus deltoides Bartr.) based agroforestry system” was carried out at Punjab Agricultural University, Ludhiana and KVK, Roopnagar (Experiment I) during 2017- 18 and 2018-19. Experiment I consisted of crop establishment methods of wheat viz. two methods of sowing (line sowing and broadcasting) and two seed rates (100 kg/ha and 125 kg/ha) along with five weed control treatments viz. carfentrazone 20 g/ha, pinoxaden 50 g/ha, clodinafop+metribuzin 270 g/ha, weed free check and weedy check in factorial RBD with 3 replications. The age of the poplar tree was 4 years in Ludhiana with 12.4 m height and 15.8 cm diameter at breast height (dbh) while at Roopnagar the age of the plantation was 3 years with an average 9.3 m height and 12.1 cm dbh during 2018-19. Experiment II consisted of planting geometry viz. two poplar spacings (8m×2.5m and 7m×3m, paired at 2.5m) and two potato planting geometry (65cm×18.5cm and 75cm×15cm) in main plots and weed control treatments (atrazine 0.250 kg/ha, metribuzin 0.350 kg/ha, paraquat 0.300 kg/ha, clodinafop+metribuzin 0.270 kg/ha, weed free check and weedy check) in sub-plots with three replications in split-plot design. The age of the poplar here was 6 years old with 19.6 m height and 26.5 cm DBH during 2018-19. The results of experiment I revealed that line sown wheat (38.0-42.8 q/ha) with higher seed rate of 125 kg/ha (37.3-41.5 q/ha) under weed free check (40.2-45.4 q/ha) had higher yield advantages with a greater number of effective tillers, spike length, grains/ear and 1000-grain weight in comparison to broadcasted with lower seed rate wheat crop under the rest weed control treatments and it was statistically at par with clodinafop+metribuzin 270 g/ha (36.5-41.3 q/ha) applied treatments. The application of clodinafop+metribuzin 270 g/ha effectively reduced the weed count and biomass. In experiment II, poplar spacing of 8m×2.5m (263.5-272.0 q/ha) and potato planting geometry of 75cm×15cm (256.6-262.1 q/ha) recorded significantly higher potato tuber yield. Among the weed control treatments weed free check (294.9-302.9 q/ha) was at par with clodinafop+metribuzin 0.270 kg/ha (289.4-293.2 q/ha) in terms of potato tuber no., weight/hill and yield. These treatments have exhibited a higher weed smothering effect thereby subsequently reduced the weed seed bank and seed recruitment in both the experiments. All the herbicides used in both the experiments temporarily reduced the microbial population and their activity upto 90 days of application and then it recovered till harvesting of the intercrops.
