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ThesisItem Open Access Antimicrobial Potential of Endophytes from Aerial Roots of Tinospora cordifolia (Willd.) Hook. F. &Thomson(Sher-e-Kashmir University of Agricultural Sciences & Technology of Jammu, 2023-11-09) Koushal, Diksha; Singh, BrajeshwarDetailed studies were conducted on evaluating antimicrobial potential of endophytes from aerial roots of Tinospora cordifolia in the Division of Microbiology, Faculty of Basic Sciences of Sher-e- Kashmir University of Agricultural Sciences and Technology, Jammu. Healthy samples of aerial roots of Tinospora cordifolia were collected from IIIM Research Farm, Chatha with GPS location (L. 32.6630276, A. 74.8145734). A total of six bacterial and five fungal endophytes were isolated from collected samples of aerial roots. Endophytes were identified on the basis of their morpho-cultural and microscopic characteristics. Identification confirmed that most of the bacterial endophytes belong to Genus Bacillus and fungal endophytes that to Genus Fusarium. Endophytes were screened for their antimicrobial potential against human pathogens: Escherichia coli, Klebsiella pneumoniae (MTCC 109) and animal pathogens: Escherichia coli (MTCC 1697), Staphylococcus aureus (MTCC 96) and Pseudomonas aeruginosa (MTCC 741). BI-2 and BI-5 have shown significant antimicrobial activity against the test pathogen, Escherichia coli (MTCC 1697) and Staphylococcus aureus (MTCC 96). BI-2 have shown maximum zone of inhibition of 8.44mm followed by BI-5 which showed an inhibition zone of 8.33mm against Escherichia coli (MTCC 1697). When screening was done against Staphylococcus aureus (MTCC 96), BI-5 showed maximum zone of inhibition of 7.54mm followed by BI-2 which showed a zone of inhibition of 7.30mm. Potent endophytes BI-2 (Ligilactobacillus salivarius) and BI-5 (Bacillus altitudinis) were processed and identified by IMTECH, Chandigarh using the MALDI-TOF mass spectrometry and 16s rRNA sequencing respectively. This study concluded that aerial roots of Tinospora cordifolia does have an immense biodiversity of endophytic microfauna possessing antimicrobial potential which could be further exploited for industrial applications and drug production.ThesisItem Open Access Studies on Medicinal Properties of Endophytes Isolated from Black Turmeric (Curcuma caesia Robx)(Sher-e-Kashmir University of Agricultural Sciences & Technology of Jammu, 2023-11-13) Kour, Ramanpreet; Singh, BrajeshwarCurcuma caesia is a medicinal herb in India commonly known as kali haldi. It belongs to family Zingiberaceae. C. caesia is a rare and unique member of the ginger family known for its distinctive black color and medicinal properties. It is a perennial herbaceous plant native to Northeast India and Southeast Asia. It has also been ranked among the 32 important plants by NMPB. In this study, samples of healthy rhizomes were collected from Marh area of Jammu district. During isolation, five bacterial and three fungal isolates were obtained from rhizomes of C. caesia. Endophytes were identified on the basis of morpho-cultural and microscopic characteristics. Endophytes were screened for their antimicrobial potential using the agar well diffusion method against animal and human pathogens. During the study, it was recorded that the fungal endophyte BTF-1 showed significant zone of inhibition of 8.23mm against Pseudomonas aeruginosa (MTCC 741) followed by BTF-3 which had zone of inhibition of 7.00mm. When screening was done against Klebsiella pneumoniae (MTCC 109), BTF-1 showed maximum zone of inhibition of 6.23mm followed by BTF-3 that had zone of inhibition of 6.20mm. Bacterial endophyte BT-2 showed a zone of inhibition of 4.44mm against Escherichia coli (animal pathogenic) and BT-3 had a zone of 5.82mm against Staphylococcus aureus. When screened against Klebseilla pneumoniae, BT-4 showed zone of inhibition of 6.66mm. From this study it was concluded that rhizomes of C.caesia retain ample biodiversity within that might be the reason for its medicinal values. As indicated in this work, potential fungal and bacterial endophytes can be further exploited for industrial applications and drug development.ThesisItem Open Access Study on plant growth promoting and antagonistic activities of rhizobacteria isolated from urad bean (Vigna mungo)(Sher-e-Kashmir University of Agricultural Sciences & Technology of Jammu, 2023-11-09) Kumari, Vinta; Dutta, UpmaThe availability of a reliable biofertilizer is essential for achieving high yields at a low cost. In the present study, rhizospheric soil and root nodules of Urad bean were collected from ACRA, Dhiansar. Isolation was done by serial dilution method. Ten isolates were isolated from rhizospheric soil and root nodules. The isolates were screened for PGP activities under in vitro condition viz, phosphate solubilizing, nitrogen producing activity, ammonia producing activity, HCN production and indole production test and for antagonistic activities against Fusarium sp. and Colletotrichum sp. using agar well diffusion method. Result showed that out of 10 isolates,5 isolates showed significant results for PGP and antagonistic activities. I-1, I-3, I-4, I8, I-10. These potential isolates were identified by Gram staining method and biochemical test. All the isolates were identified as Gram-negative bacteria. These isolates were tested biochemically and it was observed that isolates I-3, I-4 and I-10 had resemblance to the genus Pseudomonas and I-1 and I-10 had resemblance to the genus Xenorhabdus. Further, these isolates were evaluated under in vivo study to see their efficacy on growth parameters of urad bean. Seven treatments were carried out in two sets. In first set of treatment T0 is taken as control T1 is taken as inoculated control i.e., Fusarium sp. was inoculated to pot soil with no seed treatment. T2, T3, T4, T5 and T6 were inoculated with Fusarium sp. and seed was treated with potential isolates I-1, I-3, I-4, I8 and I-10 respectively. In second set of treatment same treatment was taken but the soil was inoculated with Colletotrichum sp. Out of seven treatments, two treatments i.e., T4 and T6 in which seeds were treated with I-4 and I-10 showed the best results in terms of fresh root weightroot length, shoot length, no. of pods. This study concludes that these isolates were used as bio-fertilizers for improving plant growth and used as antifungal against Fusarium sp. and Colletotrichum sp.ThesisItem Open Access Study on antimicrobial and antioxidant properties of peel and seed extract of Carica papaya(Sher-e-Kashmir University of Agricultural Sciences & Technology of Jammu, 2023-10-25) Sharma, Vasundra; Dutta, UpmaCarica papaya Linn. is a tropical, succulent and delicious fruit which belongs to the family Caricaceae and genus Carica. Since ancient times, the entire plant including peel, seeds, leaves etc. has been used as traditional medicine. Papaya peel and seeds are rich in antioxidants and have many pharmacological properties like antimicrobial, anticancer, antifertility,anti- inflammatory and many more. In the present study, papaya peel and seed samples were collected from the local market of Jammu district of J&K. Their fine powder was used for extraction using ethanol as solvent to obtain crude extract. These extracts were used for phytochemical analysis as well as for evaluation of antimicrobial and antioxidant activities. The antimicrobial properties were assessed using agar well diffusion method and the results revealed that both the extracts showed inhibition against four human pathogens (Escherichia coli, Bacillus cereus, Klebsiella pneumonia and Staphylococcus aureus) at 100 µg concentration followed by 80 µg concentration except for E. coli in peel extract. The minimum inhibitory concentration was conducted using macro broth dilution method and the result showed that MIC of seeds was higher than that of peel against the test pathogens. The qualitative phytochemical analysis showed that phytochemicals including carbohydrates, flavonoids, quinones, glycosides, terpenoids, phenols and steroids were present whereas tannins, saponins, alkaloids and phytosteroids were absent in both the extracts. The total phenolic content and total flavonoid content were more in seed extract than peel extract. The antioxidant potential of papaya peel and seeds were screened using DPPH and β-Carotene /Linoleic acid test in which peel extract showed higher inhibition as compared to seed extract. There was positive correlation between total phenol content and total flavonoid content with DPPH free radical scavenging activity and β-Carotene /Linoleic acid test for both the extracts and also between TPC and TFC of papaya peel and seeds with MIC’s of the test pathogens. This study concludes that the papaya waste can be a good source of antimicrobial and antioxidant agents. However, further research should be done to identify the active components responsible for these activities.ThesisItem Open Access Studies on bio-degrading potential of fungi isolated from kitchen waste(Sher-e-Kashmir University of Agricultural Sciences and Technology of Jammu, 2022-11) Chib, Muskaan; Dutta, UpmaThe non-edible portions of food that are wasted during handling, processing, or after consumption are referred to as "kitchen waste." It contains unavoidable food wastes from refrigerators and food preparation areas, as well as garbage from homes, businesses, institutions, and other sources. Due to its high moisture concentrations, kitchen waste is difficult to handle using conventional methods, such as incineration. However, it is crucial to find an alternative to securely dispose of organic waste since landfills and incinerators are closing at an alarming rate. Efficient decomposition of food waste may be achieved by a variety of fungus that uses cellulose, proteins, lipids, and carbohydrates to fulfill their energy needs. Therefore,it is possible to accelerate the biodegradation of kitchen trash by establishing consortia of these fungus. In the present study, eight fungal isolates were isolated from kitchen waste samples collected from the canteen and mess of SKUAST-Jammu. The screening of the isolates was done by measuring zone of clearance and halo formation around the growth on specific media viz. tributyrin agar, skim milk agar, carboxy methyl cellulose agar, and starch agar and five potential isolates were screened out followed by their morphological and microscopic identification. The compatibility of the potential isolates was checked by dual culture method and a consortium was prepared by the inoculation of compatible isolates in Potato dextrose broth followed by incubation at 28C for 48 hrs. 450 g of kitchen waste was used in different trials separately at different pH (5.7, 6.5, and 7.5) and temperatures (25C, 30C, and 40C), and the study showed that at 30C temperature with pH 5.7, weight reduction was highest (90.80%) The effect of consortium on pre-treated kitchen waste (acid and thermal treatments) was also studied and the results revealed that the pre-treated kitchen waste with consortium significantly degraded the waste over control and among them, acid treatment of kitchen waste with consortium showed the maximum degradation of kitchen waste (37.47%). This study concluded that the consortium of potential biodegrading fungi enhanced the biodegradation process of kitchen waste.ThesisItem Open Access Bioprospecting Actinobacteria in quest of industrial application(Sher-e-Kashmir University of Agricultural Sciences and Technology of Jammu, 2022-09) Vironika; Singh, BrajeshwarAn investigation entitled “Bioprospecting Actinobacteria in quest of industrial application” was conducted in the Division of Microbiology, Sher-e-Kashmir University of Agricultural Sciences and Technology of Jammu, to study the antibacterial and biopigments potential of actinobacteria. Actinomycetes are widely distributed throughout the globe, and show overwhelming excellence in the industrial, pharmaceuticals, food and textile sector. In this study 33 actinobacteria cultures were isolated, purified for further studies from forest area of Sidhra, Miran Sahib and Bhaderwah-I & II of Jammu region. Morphological study revealed that isolated cultures had white, silver-grey, off-white colonies, pin head size, rough margin, powdery consistency, were raised and embedded; characteristics similar to that of actinobacteria on YMA media. 33 actinobacteria were screened by agar well diffusion method against four test human pathogens Pseudomonas aeruginosa, Salmonella typhimurium, Staphylococcus aureus and Escherichia coli. 26 isolates had indicated antibacterial activity viz. 10 from Sidhra (SA01, SA05, SA07, SA08, SA10, SA13, SA23, SA82, SA92, SA95); 02 from Bhaderwah-I (SB01, SB02), 09 from Bhaderwah-II (SD02, SD03, SD04, SD06, SD09, SD12, SD13, SD16, SD19) and 05 from Miran sahib (SC01, SC04, SC05, SC06, SC07). 05 potential isolates SA92, SD04, SD12, SD09 & SD16 were selected for further studies as they had antibacterial activities as well as biopigments production potential. Water extract of SA92 and SD04 used for dying cotton fabric, that produced brown and purple pigment respectively that were stable against heat and sunlight, indicating industrial application potential. Substrate utilization by these 05 isolates indicated that they are member of Streptomyces group.ThesisItem Open Access Studies on Vitamin B12 production by Lactobacillus species isolated from milk products(Sher-e-Kashmir University of Agricultural Sciences and Technology of Jammu, 2021-12) Sharma, Shruti; Singh, BrajeshwarAn investigation entitled “Studies on production of Vitamin B12 by Lactobacillus species isolated from milk products” was conducted in the Division of Microbiology, Sher-e-Kashmir University of Agricultural Sciences and Technology of Jammu, to study the capability of mutants generated out of UV and EMS mutagens of native Lactobacillus isolates, for Vitamin B12 production potential. In this study 8 isolates of Lactobacillus were isolated from curd and kaladi. Screening and Quantification of isolates was done on Vitamin B12 assay medium. The Vitamin B12 production by Lb-7 was 0.39 mg/g DCW. Lb-7, the fastest growing strain was selected for strain improvement, that was done by UV and EMS mutagens. Based on performance during screening, UV mutant was taken up for fermentation optimisation studies. The Vitamin B12 production by UV mutant was 0.63mg/g DCW. Three fermentation parameters i.e temperature, pH and inoculum load were optimized for mutant to enhance Vitamin B12 production. The Vitamin B12 production at 25°C was 2.05 mg/g DCW ,5.5 pH was 1.55mg/g DCW and 108 inoculum load was 1.53mg/gDCW. Best results were recorded at 25°C, 5.5 pH and 108 cfu/ml of microbial load. An increase in temperature, pH and inoculum load caused decrease in Vitamin B12 production due to death of cells.ThesisItem Open Access Augmentation of kitchen waste by resident biodegrading bacteria(Sher-e-Kashmir University of Agricultural Sciences and Technology of Jammu, 2022-02) Attri, Harkiran; Dutta, UpmaKitchen waste is renewable, low-cost, and made up of organic ingredients viz. carbohydrates, amino acids, peptides, fatty acids, and their esters, as well as processed foods and paper make up this group that provide bacteria with the nutrients they need to thrive. Improper handling of kitchen waste can cause environmental pollution and can have a substantial impact on public health. Enzymes produced by bacterial consortium (amylase, lipase & protease, cellulose, lactose fermenting bacteria) is essential for decomposition of kitchen waste. Enzymes from microbes have a wide biochemical diversity, and are easy to manipulate genetically. Effective microorganisms play an important role in composting process by enhancing the pace of bio-degradation. In the present study twenty bacterial isolates were isolated from the kitchen waste sample collected from Urja girl’s hostel SKUAST-Jammu. Biodegrading ability of these isolates were screened by measuring zone of clearance and halo formation around the growth on specific media viz. starch agar, skimmed milk agar, MacConkey agar, and Congo red agar. Nine potential isolates were screened out for further studies. Morpho-cultural and biochemical identification of potential isolates were done onas per Bergey’s Manual of Determinative Bacteriology. The compatibility of the potential isolates was studied by cross streak method and consortium was prepared by the inoculation of compatible isolates in nutrient broth. 500 g of kitchen waste was used in different trials separately at different pH (5.7, 6.5 and 7.5) and temperatures (37ºC, 40ºC and 45ºC) and the study showed that at 45ºC temperature with pH 7.5 among all the trials showed highest weight reduction (75.60%). The effect of consortium on pre-treated kitchen waste (acid and freeze-thaw treatments) was done and the results revealed that the pre-treated kitchen waste with consortium significantly degraded the kitchen waste over control but among them, acid pre-treatment of kitchen waste with consortium showed the maximum degradation of kitchen waste (21.00%). This study concluded that the consortium of resident potential bio-degrading bacteria enhanced the bio-degradation process of kitchen waste.ThesisItem Open Access Studies on bioethanol production from rice straw by Saccharomyces cerevisiae(Sher-e-Kashmir University of Agricultural Sciences and Technology of Jammu, 2022-02) Choudhary, Neeru; Singh, BrajeshwarAn investigation entitled ‘Studies on bioethanol production from rice straw by Saccharomyces cerevisiae’ was conducted in the Division of Microbiology, Sher-e-Kashmir University of Agricultural Sciences,with a view to study the bioethanol producing potential of locally isolated Saccharomyces cerevisiaestrain;using Rice straw as a substrate.Optimization of fermentation parameters i.e., temperature, pH and agitation to enhance the bioethanol production was also studied. Fifteen yeast strains were isolated from fruit sample (Apple, Papaya, Sugarcane, Grapes, Orange, Pineapple) and Kaladi samples collected from local markets of Jammu and Udhampur district respectively. Potential yeast isolates were identified on the basis of morphological and microscopic characteristics which were similar to that of MTCC 170 culture procured from IMTECH-Chandigarh. Out of 15 isolates, 14 show a positive change in the color of screening medium indicating the production of ethanol. S-9 culture was further selected based on its highest bioethanol producing ability of 17.88 mg/mL at 24 h of fermentation on YEPD medium and hence chosen for the fermentation of rice straw hydrolysate.Rice straw was subjected to alkaline and acidic pretreatment which was followed by enzymatic hydrolysis.Ethanol Fermentation of the total Rice Straw Hydrolysate obtained after pretreatments using S-9 culture revealed maximum ethanol concentration of 12.26 mg/mL at 24h havingfermentation efficiency of 69.5%, 50.7% of sugar conversion occur during the fermentation process and volumetric productivity of 0.1702 g/L/h. Ethanol content per gram of sugar utilized was estimated to be 0.168g/g. Further, S-9 culture show the maximum efficiency for bioethanol production on YEPD medium at temperature of 30ºC, pH of 5.6 and under static conditions.