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ThesisItem Restricted 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 Evalution of Plant Growth Promoting Rhizobacteria As potemtial Biofertilizer For Broccoli ( Brassica Oleracea L.)(Punjab Agricultural University, Ludhiana, 2022) Sharma, Kavia; Jupinder KaurThe present study was carried out with the aim to develop a plant growth-promoting rhizobacterial inoculant for improving the growth and yield of broccoli. A total of 24 bacteria were isolated from rhizosphere of broccoli collected from the fields of Punjab Agricultural University, Ludhiana and village Sangrur, Mansa, Barnala and Patiala. Biochemical characterization of these rhizobacterial isolates showed that most of these were oxidase, urease, indole, MR and catalase positive and were involved in starch hydrolysis and gelatin liquefaction whereas negative for citrate and VP test. These isolates were screened for multifarious plant growth-promoting traits viz. solubilization of phosphate and zinc, production of IAA, siderophore, ammonia, HCN and growth on nitrogen free medium. Out of 24 isolates, 17 isolates showed solubilization of phosphate on modified Pikovskaya agar with solubilization index ranging from 0.5-4.5 cm and 53.63-178.6 μg/ml in liquid medium. Eleven isolates showed solubilization of zinc oxide (solubilization index: 0.4-1.9 cm), five isolates showed solubilization of zinc carbonate (solubilization index: 0.3-1.2 cm) and zinc phosphate (solubilization index: 0.1-2.5 cm). Siderophore production was exhibited by fourteen isolates (0.3-1.4 cm). All the isolates were positive for auxin (without tryptophan: 2.04- 13.90 μg/ml; with tryptophan: 10.05-32.04 μg/ml) and ammonia production as well as ammonia excretion (1.803.65 μg/ml). The luxuriant growth on different nitrogen free medium was observed for all the isolates and higher HCN production was observed in three bacterial isolates. Out of these 24 isolates, two potential bacterial isolates RP4 and RP5 (which were named as CBF 2) were selected on the basis of PGP traits for evaluation of their potential to promote growth and yield of broccoli under pot as well as under field conditions. In green house experiment, these isolates when used synergistically showed a significant improvement in soil enzymatic activities (viz. dehydrogenase: 26.13 µg TPF formed h-1 g-1 of soil, alkaline phosphatase: 14.02 µg PNP formed h-1 g -1 of soil and urease: 370.80 µg urea hydrolysed formed h-1g -1 of soil) and plant growth parameters (plant height: 45.7 cm, root length: 6.3 cm, stem diameter: 3.9 cm, fresh weight of shoot :167.85 g, dry weight of shoot:35.70 g, number of leaves per plant: 26, days taken to visible curd:56 days, head compactness: 1.36 cm, number of heads/plant: 4 heads, mean head weight: 87 g and floret stalk length: 12.8 cm) over the control as well as other bioinoculants. The field evaluation of these bacterial isolates as consortium along with FYM and inorganic fertilizers was carried out using Palam samridhi variety of broccoli. The combined application of bacterial strains (RP4 and RP5) and FYM resulted in significantly higher soil microbial population and enzyme activities. Significantly higher available nitrogen (137.30 kg ha-1), available phosphorus (34.21 kg ha-1) and available potassium (170.15 kg ha-1) were also observed in treatment having FYM and bacterial inoculants (N75FYM+N25 + CBF 2). Significantly higher plant nutrient uptake of nitrogen (98.87 kg ha-1), phosphorus (24.84 kg ha-1), potassium (182.27 kg ha-1) was observed in (N75FYM+N25+ CBF 2) and higher uptake of micronutrients viz. zinc (75.5 mg/kg), iron (172.16 mg/kg) and manganese content (71.11 mg/kg) were found to be significantly higher in treatment (N100FYM+ CBF 2) having synergistic application of organic and bio-fertilizers. The various growth attributes of broccoli viz. plant height: 42.1 cm, root length: 11 cm, stem diameter: 4.9 cm, fresh weight of shoot: 214.20 g, dry weight of shoot:38.18 g, number of leaves per plant: 29, days taken to visible curd:44 days, head compactness: 1.11 cm, head yield: 153.15 q/acre, number of heads/plant: 6 heads, mean head weight: 88 g and floret stalk length: 12.8 cm was observed to be statistically higher in treatment T10 with combined application of FYM and bacterial isolates (N100FYM+ CBF 2). The study has led to the selection of a plant growth-promoting bacterial consortium as a potential bacterial inoculant for improving the growth, yield and nutrient content in broccoli.ThesisItem Restricted Bioprospecting production of fermented type II low fodmap (fermentable oligosaccharides, disaccharides, monosacchrides and polyols) sourdough(Punjab Agricultural University, 2023) Khanna, Rahul; Arora, RichaFermentable oligosaccharides, disaccharides, monosaccharides and polyols (FODMAPs) in the food are short chain carbohydrates which are poorly absorbed and easily fermented in the large intestine by colon micro-organisms, thereby, producing acid and gas. These FODMAPs adversely affect human health leading to irritable bowel syndrome (IBS). In this context, sourdough technology can be exploited to reduce the FODMAP levels in food and alleviate the symptoms of IBS. In the present study, three yeast cultures, viz. Saccharomyces cerevisiae, Clavispora lusitaniae and isolate YB1 were compared for dough raising capacity, where yeast isolate YB1 was screened for maximum dough raising capacity (9.20 %). The screened isolate was characterised as Meyerozyma guillermondii by 18SrRNA gene sequencing and analysed by BLAST. Furthermore, the fermentation parameters for maximum dough raising capacity were optimised as inoculum size 13.6 % (2.4 x cfu/ml) and fermentation time 25.9 h using response surface methodology. After optimisation, fermentation of all- purpose flour using YB1 isolate resulted in reduction of total sugars (84.1 %), reducing sugars (79.8 %), glucose (60.2 %), fructose (88.8 %), lactose (39.3 %), sucrose (98 %) and raffinose (30.5 %). Apart, from sugar reduction, the fermented sourdough was found to have enhanced total antioxidants by 42.3 % and decrease in concentration of anti- nutritional compound i.e. phytic acid (16 %) after 25 h. Colour change by the fermentation process was found to be non- significant. Rheological characteristics of the sourdough, viscosity and shear strain decreased by 60 and 50 % respectively, thereby, increasing the porosity of the product. A total of 34 volatile compounds were detected using GC-MS with major contribution of butanoic acid, 2,3- butanediol tetracontane, n-hexadecanoic acid, octadecanoic acid which enhances aroma and shelf life of the fermented food. The present study demonstrates the potential of yeast isolate YB1 for production of low FODMAP sourdough fermented products using all-purpose flour as a promising alternative food for IBS patients.ThesisItem Embargo Evaluation of microalgal-bacterial consortia for waste water treatment(Punjab Agricultural University, 2023) Poonam; Sharma, Shiwani GuleriaThis study investigated the bioremediation abilities of microalgae and bacteria for the treatment of domestic waste water. Two microalgal viz. PL1 and PL2 and eight bacterial strains viz. B1(L1), B2(L1), B3(L1), B4(L1), B1(L2) B2(L2), B3(L2) and B4(L2) were isolated form indigenous population i.e L1 & L2. Both the microalgae isolates had a long exponential phase and continued to remain in this phase till the end of experimental growth period i.e. 30 days. Log phase of all bacterial isolates were observed at 60 hours of incubation except B1(L1) (48 hours). After screening, two microalgae i.e. PL1 & PL2 and four bacterial isolates were selected for efficient wastewater treatment. B1(L1), B3(L1), B2(L2) and B4(L2) were observed to be efficient in wastewater treatment than other bacteria under study. Coexistence of each strain with other was seen without any trace of growth inhibition which represented their compatibility. From the total of 29 tested consortia, most potential consortium was selected based on the treatment efficacy after 15 days of treatment. Developed consortium no. 29 i.e. PL1+PL2+B4(L2) was selected for domestic wastewater treatment as it showed significant percent reduction among all combinations i.e. BOD (82.9%), COD (77.77%), EC (53.8%), TDS (74.3%),Total nitrogen (71.05%) and Total Phosphorus (69.25%). Higher percent reduction in BOD (88.90%), COD (82.35%) was recorded with selected consortia 29 when treatment provided with source of aeration in 30 days of treatment. SEM observations revealed that several bacterial cells were attached to microalgal cells over 10 days of incubation and it led to the conclusion that microalgae PL1 & PL2 and bacteria B4(L2) can live together and efficiently treat wastewater. Molecular identification of members of consortia 29 was performed. It was revealed after 18S rRNA sequencing PL1 was identified as Chlorella variabilis, PL2 was tentatively identified as Desmodesmus and after 16S rRNA sequencing B4(L2) was identified as Paracoccus sp.. Sequence of PL1 and B4(L2) was submitted to NCBI GenBank with accession no. OR122520.1 and OR467496, respectively. Thus, present study concluded that microalgae and bacteria have more potential to treat wastewater when they were applied in form of consortia. The selected developed consortia no. 29 showed highest percent reduction in all physiochemical parameters with source of aeration in 30 days of wastewater treatment. Nevertheless, the assessment of polluted water composition should be completed prior to the designing and application of microalgae or cyanobacteria based remediation technology, since the pollutant removal efficiency is found to be essentially species dependent.ThesisItem Restricted Effect of slow release urea nano fertilizer on soil microflora and yield of direct seeded rice (Oryza sativa L.)(Punjab Agricultural University, 2023) Yashika; Kalia, AnuThe present work aimed to study the effect of a slow release urea nanofertilizer on soil microflora and yield of direct seeded rice (Oryza sativa L.). A soil column study indicated slow-release pattern of the urea from the nano-urea product varied for the soil samples having three different textures (low, medium and heavy).The leachate analysis revealed release of ammonical and nitrate nitrogen from the synthesized product. Further, the effect of the nanourea fertilizer on soil microflora study identified that the viable cell counts for a variety of soil microbes were not negatively affected on application of different doses of nanourea fertilizer. The soil urease, protease, and dehydrogenase enzyme activities exhibited significant increase in nano urea (100% RDN) treatment in light textured soil. A field experiment was performed to observe the effect of nano urea fertilizer on the growth and yield of direct seeded rice. An increase in the plant height, shoot and root fresh weight and dry weight was recorded for different doses of Nitrogen applied through nano urea and conventional urea N-fertilizer level for plants sampled after every 30 days after sowing till three months of the growth. The maximum plant growth was recorded for treatments which received 100% of the recommended N-fertilizer. Likewise, the yield attributing traits particularly the number of filled grains and thousand grain weight increased on application of 100% Recommended dose of N through nano urea. The soil chemical parameters were recorded to be non-significantly altered for plant and grain nutrient content (NPK content) at nano-urea fertilizer (100% RDN). Non-significant effect was recorded for the soil microbial viable cell count on different doses of nano-urea fertilizer. These results indicate the use of nano-urea fertilizer as a possible alternative for conventional urea fertilizer.ThesisItem Embargo Characterization and industrial application of oxidative/hydrolytic enzymes of Pleurotus florida (Mont.) Sing. and Calocybe indica (Pur. & Chan.)(Punjab Agricultural University, 2023) Manmeet Kaur; Sharma, ShivaniPleurotus florida and Calocybe indica are the edible mushrooms that have been widely accepted due to their ability to grow on a variety of substrates and possess the potential to produce oxidative and hydrolytic enzymes. The present work involved the study and characterization of oxidative/hydrolytic enzymes of P. florida and C. indica for their potential applications in alcohol fermentation, dye decolorization, biobleaching and mushroom production. The oxidative and hydrolytic enzymes extracted from P. florida and C. indica were estimated at different mycelial growth stages for enzyme production. The intracellular and extracellular enzyme activity of P. florida and C. indica increased with time and maximum activity of lignocellulolytic enzymes was found to be on 14th day of incubation. In both mushrooms, ligninolytic enzyme activity increased during substrate colonisation but quickly decreased during fruiting body development. On the other hand, P. florida and C. indica showed relatively modest hydrolase activity during substrate colonisation. The activity of hydrolytic enzymes increased dramatically during primordial formation and peaked at the fruiting body stage. The purification of laccase, lignin peroxidase, manganese peroxidase and endoxylanase from the fruiting body of P. florida resulted in maximum purification fold of 21.49, 17.73, 16.81 and 12.78 with yield of 24.98%, 20.60%, 19.53% and 14.86%, respectively. However, the purification of laccase, lignin peroxidase, manganese peroxidase and endoxylanase from the fruiting body of C. indica resulted in maximum yield of 21.36%, 18.92%, 17.37% and 16.62% with purification fold of 25.6, 25.12, 20.88 and 19.97, respectively. The SDS-PAGE of the purified enzyme laccase and lignin peroxidase isolated from P. florida showed a single prominent band at 66 kDa and 55 kDa respectively. The SDS-PAGE of the purified enzyme laccase and lignin peroxidase isolated from C. indica showed a single prominent band at 64 kDa and 47 kDa, respectively. FTIR spectra of the purified enzymes indicated a secondary structure that reflected the amide I and amide II bands, respectively. The pretreatment of paddy straw and wheat straw with P. florida, C. indica and their ligninolytic enzyme resulted in decrease in lignin and hemicellulose content, respectively during the incubation period of 30 days. However, the decrease in cellulose content occurred during the pretreatment of substrates with fungus while relative increase in cellulose content during the treatment with ligninolytic enzymes was observed over the incubation period of 30 days. Under optimized conditions, saccharification of biologically pretreated paddy and wheat straw resulted in release of 0.415 and 0.389 g/gds reducing sugars, respectively. The fermentation of biologically pretreated and commercial cellulase saccharified paddy and wheat straw hydrolysate resulted in 0.129 and 0.119 g/g ethanol, respectively. The crude enzyme extract of P. florida and C. indica were able to degrade RBBR (25.74 %, 22.06%) and Amido Black (19.76%, 17.58%) dyes maximally after 96 hours of incubation at 30°C and pH 7.0. The paddy and wheat straw was treated with a ligninolytic crude enzyme, which stimulated faster mushroom growth and fructification. The present study thus revealed that expression of biosynthetic potential of P. florida and C. indica is highly dependent on the method of fungi cultivation. These ligninolytic enzymes showed a unique profile in terms of versatility, greenness, pollutant removal and efficiency in lignin degradation for the exploitation and valorization of agro-wastes.ThesisItem Restricted Prospecting plant growth promoting and endophytic bacteria for improving growth and yield of sunflower (Helianthus annuus L.)(Punjab Agricultural University, 2023) Kiranjot Kaur; Vyas, PratibhaThe current study was carried out with the aim to develop a plant growth-promoting bacterial inoculant for improving the growth and yield of sunflower. A total of 64 morphologically distinct bacteria were isolated from leaves, stem, roots and rhizosphere of sunflower collected from the fields of Punjab Agricultural University, Ludhiana. The isolates were screened for multifarious plant growth-promoting traits viz. solubilization of phosphate and zinc; production of auxins, siderophore, ammonia, hydrogen cyanide, and growth on nitrogen free medium. Thirty-three bacterial isolates showed solubilization of phosphate with solubilization index ranging from 2.7 to 3.8 by rhizospheric bacteria and 2.4 to 5.4 by endophytic bacteria on modified Pikovaskaya agar. In liquid medium, phosphate solubilization ranged from 3.6 to 171.6 μg/ml by the rhizospheric bacteria and 1.1 to 889.5 μg/ml by endophytic bacteria. Thirty isolates showed zinc solubilization, 25 isolates showed siderophore production while 40 isolates showed luxuriant growth on different nitrogen free media. All the isolates were positive for auxin and ammonia production but negative for hydrogen cyanide production. Based on multiple plant growth-promoting traits, nine potential bacterial strains were tested for growth promotion of sunflower in pots. The isolates showed a significant increase in growth, chlorophyll content and N, P and K contents of plant and soil samples with the bacterial inoculations over the uninoculated control. Three isolates showing the highest growth promoting activity were identified by phenotypic features, biochemical tests and MALDI-TOF-MS analysis as Pseudomonas aeruginosa SRB1, Advenella kashmirensis SLE1 and Brevendimonas dimunata SLE15. The field evaluation of three potential bacterial strains Pseudomonas aeruginosa SRB1, Advenella kashmirensis SLE1 and Brevendimonas dimunata SLE15 and their consortium was carried out with and without 75 and 100 per cent recommended dose of fertilizers for sunflower. The highest growth promotion, nutrient content and yield enhancement was recorded with the treatment AkSLE1 + BdSLE15 + PaSRB1 with 100 per cent RDF which was statistically not different from that obtained with the treatment AkSLE1 + BdSLE15 + PaSRB1 with 75 per cent RDF, indicating the potential of these three PGPB strains in saving around 25 per cent N, P and K fertilizers for sunflower. The study has led to the selection of a plant growth-promoting bacterial consortium as a potential bacterial inoculant for improving the growth, yield and nutrient content in sunflower.ThesisItem Restricted Evaluation of bacterial consortium to boost in-situ degradation of Sesbania sp. to improve soil fertility, plant growth and yield of maize crop(Punjab Agricultural University, 2023) Khipla, Neha; Sharma, PoonamPresent research work was aimed at evaluating bacterial consortium for boosting in-situ decomposition of Sesbania (green manure) to improve soil health, growth and yield of Kharif maize (var. PMH1). Three cellulolytic bacterial cultures viz. CD2, CD9 and CD14, identified through 16S rDNA sequence similarity were published at GenBank under strain name Lysinibacillus sp. Strain JNcd2 (OM721941.1), Glutamicibacter mishari Strain JNcd9 (OM723742.1) and Paenibacillus sp. Strain JNcd14 (OM727120.1), respectively. The bacterial strains were positive for various plant growth promoting traits such as ammonia production and excretion, phosphorus and zinc solubilization, starch hydrolysis, IAA and siderophore production. All the bacterial strains were tested positive for ligno-cellulolytic enzyme production in both qualitative and quantitative assays; and thereby, designated as Ligno-cellulolytic Bacterial Consortium (LBC). Primarily, the effect of LBC inoculation on decomposition and nutrient mineralization of Sesbania and Sesbania + rice straw was studied through 60 day laboratory incubation study during Kharif 2018. LBC inoculation increased weight loss rate of Sesbania by 17.7 % and of Sesbania + rice straw by 11.6 % over uninoculated control, till 16th day of decomposition. Total carbon content and C:N ratio of both the organic inputs was significantly lower in LBC inoculated treatment over uninoculated control, till 30th day of decomposition. Highest mean soil viable population of total bacteria and cellulolytic bacteria was recorded in Sesbania + LBC treatment, total fungi and nonsymbiotic diazotrophs in Sesbania + rice straw treatment; while, PSB and actinomycetes in Sesbania + rice straw + LBC treatment. Soil respiration, organic carbon, ammonical and nitrate nitrogen available phosphorus and potassium content was higher by 19.2 %, 6.8 % 15.2 %, 2.7 %, 1.6 % and 5.4 % in Sesbania + LBC treatment, and by 4.1 %, 0.8 %, 5.3 %, 3.4 %, 2.7 % and 4.9 % in Sesbania + rice straw + LBC treatment, respectively than uninoculated controls, at 60th day of decomposition. Field studies were conducted for two consecutive Kharif seasons (2019 and 2020) to study the effect of LBC inoculation with different sole inorganic and integrated fertilizer treatments on soil bio-chemical properties, plant growth, nutrient acquisition and yield of maize. Before sowing of maize, soil treatment with Sesbania + LBC showed highest fertility status over bare, than sole LBC and Sesbania treatment. In maize rhizosphere, soil viable microbial population (total bacteria, fungi, PSB and cellulolytic bacteria) and enzymatic activities (dehydrogenase, alkaline phosphatase, urease, cellulose and invertase) remained significantly higher in treatment with 100 % N + Sesbania + rice straw mulch (RSM) + LBC, during both experimental years. However, soil actinomycetes and non-symbiotic diazotrophs were highest in 75% N + Sesbania + RSM + LBC treatment and soil amylase activity in 100 % N + Sesbania + LBC treatment. Soil chemical properties viz. electrical conductivity, organic carbon, available P and K content, were highest in 100 % N + Sesbania + RSM + LBC treatment; soil available N content in 100 % N + Sesbania + LBC treatment; and soil pH varied numerically. During both experimental years; highest plant nutrient (NPK) uptake, growth and yield attributes, grain and stover yield of maize was recorded with treatment 100 % N + Sesbania + RSM + LBC. It was concluded that integrating LBC with organic and inorganic fertilizers could ensure efficient decomposition and nutrient mineralization in soil, and thus contribute to improved soil health, growth and productivity of maize.ThesisItem Restricted Value Addition Of Volvariella Volvacea (bull.) Singer Through Mushroom Paste Formulations In Culinary Recipes(Punjab Agricultural University, 2023) Haifa; Jaspreet KaurPresent investigation was conducted to develop paste formulations of Volvariella volvacea (Paddy Straw mushroom) for its shelf life enhancement. Fruiting bodies of mushrooms harvested at egg stage were grinded to make salted, spiced and their fermented pastes. The salted paste formulation consisted of 2.5% salt, 0.5% sugar, 2ml refined oil, 1.5ml vinegar and 0.1% each of ginger and garlic while spiced paste was formulated by adding 0.5% onion, 0.1% cumin, 0.1% red chilli and 0.01% black pepper to the salted paste. To develop fermented paste formulations, salted and spiced paste was fermented using Lactobacillus rhamnosus GG for 2 days at 35±2°C. The unfermented mushroom spiced and salted paste was preserved with sodium benzoate @ 0.065% while fermented formulations were kept devoid of the any chemical preservative. The paste formulations were packed in glass jars and retort bags, and thereafter treated in boiling water bath for one hour. In organoleptic evaluation, the spiced fermented mushroom paste scored (8.3) significantly higher from the salted fermented formulation (7.7). Nutritional analysis of all the formulations revealed that spiced fermented mushroom paste contained highest content of crude protein (23.67%) and moisture content (36.72%) while lowest content of ash (3.01%), fat (0.51%), crude fiber (7.92%) and carbohydrates (28.17%). Highest fat (0.92%) was recorded in salted formulation. The overall energy was recorded highest (233.23J) in salted mushroom paste which was 10.04% higher than fermented spiced paste. The shelf life studies revealed that unfermented and fermented paste formulations showed spoilage on 3rd and 28th day of storage in glass jars at room temperature, as a indicated by total bacterial count of more than permissible limit of FSSAI, India. In contrast, longer shelf life of 56 days was observed for fermented mushroom paste stored in glass jars in refrigerated conditions. Variation of pH was indicator of spoilage, it was found that mushroom pastes stored in retort bags varied significantly in pH and were more prone to bacterial as well as fungal contamination irrespective of the temperature conditions. However, when stored in glass jars, the fermented (salted and spiced) mushroom pastes showed numeric variation in pH at 4ºC. The antioxidant analysis revealed that DPPH radical scavenging activity of fermented paste formulations increased significantly upto 21 days followed by gradual decrease till 56th day. While in case of unfermented paste steep decrease in DPPH activity was observed during storage. Total phenolic content of all the paste formulations was found to be decreased with storage. Above results showed that fermented spiced mushroom paste was best formulation, thus was selected for incorporation to culinary recipes. Incorporation of this mushroom paste at 25%, 10% and 45% was found best in noodles, spread and aloo gobhi sabji, respectively. It was concluded that fermentation of Volvariella volvacea spiced mushroom paste with L. rhamnosus GG culture for two days significantly enhanced its nutritional properties, antioxidants activity and shelf life upto 56 days; hence can be a good value addition to most perishable mushroom.
