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ThesisItem Open Access Effect of Halotolerant Bio-fertilizer Formulations on Production of Rice (Oryza sativa L.) in Sodic Soil of Eastern Plain Zone of Uttar Pradesh(Department of Industrial Microbiology, Jacob Institute of Biotechnology and Bioengineering, Faculty of Engineering and Technology, Sam Higginbottom University of Agriculture, Technology & Sciences, Prayagraj (Allahabad), (U.P.) India, 2019) Hafeez, Mohammad; Masih, HarisonThe present research work was conducted in the Department of Industrial Microbiology, on the topic entitled, “Effect of Halotolerant Bio-fertilizer Formulations on Production of Rice (Oryza sativa L.) in Sodic Soil of Eastern Plain Zone of Uttar Pradesh”, and the soil samples were collected from different villages of Allahabad, was analyzed for their physicochemical properties and halotolerant bacterial strains were isolated. The organisms for salt, pH tolerance, nitrogen fixing Azotobacter and phosphate solubilizing bacterial strains (PSB) were identified. The efficient strains were formulated with fly ash for bio-efficacy of carrier based bio-fertilizer was performed in pots with sodic soil during 2013 and 2014 for rice crop. The results showed that a total number of 26 efficient isolates of Azotobacter and phosphobacterial strains were isolated and identified (8 isolates of Azotobacter and 18 isolates of PSBs). These isolates survived at pH 9 and found to be tolerant up to 2% NaCl. The maximum nitrogen fixing activity (9.00 mg N/g substrate) by Azotobacter salinestris (MF144547.1) and maximum phosphate solubilizing activity (SI) = 3.80 cm) and 33.00 (μg/ml) qualitatively and quantitatively respectively by Enterobacter cloacae (MG255304) was selected for further study. Formulations of A. salinestris and E. cloacae in fly ash at 30 oC and 25 oC storage temperatures and 35% and 30% to moisture content found best conditions respectively. Shelf life study of bio-formulations stored at optimized conditions up to six months showed that the cells were gradually decreased up to six months of storage and were found maximum at initial stage (41 and 51 cfu x 108/g) respectively. Bio-efficacy studies with A. salinestris and E. cloacae formulations with different treatments showed that soil condition of pots were improved maximum by treatment T3 (A. salinestris and E. cloacae), and combination of these enhanced maximum root length (cm) (13.92%) and (14.53%), root biomass (g) (13.53%) and (19.30%), number of tillers (10.34%) and (15.62%), plant height (cm) (7.69%) and (4.00%), plant dry weight (g) (10.22%) and (15.09%), test weight (g) (5.88%) and (6.84%) and seed yield /hill (g) (8.69%) and (15.03%) were increased in first year and second year as well in comparison to control, respectively. The results of present investigation suggested the positive effect of co-inoculation of bio-fertilizer formulations that improved soil health/soil fertility which enhanced the growth and yield of rice in sodic soil. From the above findings it can be concluded that halotolerant strains of bio-fertilizer formulations in fly ash could be a better option for reclamation, growth and yield of the crop in sodic soil.ThesisItem Open Access ―Study of Erythromycin resistance genes of Staphylococcus aureus isolated from throat swab culture”(Faculty of Health Sciences Sam Higginbottom University of Agriculture, Technology and Sciences India, Allahabad-211007 2018, 2018) ABULKASIM, HALEMAH MOHAMED; Shukla, Prof. G S; Masih, Dr. HarisonThis study entitled “Erythromycin resistance genes of Staphylococcus aureus isolated from throat swab culture” was carried out in Cytogen Research and Development laboratory Indira Nagar, Lucknow, Uttar Pradesh, India from January 2016 to May 2016. The aims of this study: (1) To isolate and identify Staphylococcus aureus from throat swab samples of respondents. (2) To study the antibiotic resistance pattern of the isolates. (3) To determine erythromycin resistance methylase (erm) genes of Staphylococcus aureus by PCR and sequence technique. Out of one hundred eighty five (185) throat swab samples, 87(47%) of samples were positive for bacterial growth and 98(52.9%) were negative growth. The presence of S. aureus in throat swab culture was 47(54%) , while presence of CNS was 40(45.9). the throat swab samples were collected from students ages 17-28 years, presence of S. aureus in age group from 17-18 years was 6 (12.76%), 19-20 years was 13 (27.65%), 21-22 years was 15 (31.91%), 23-24years S. aureus was 6(12.76%), 25-26 years S. aureus was 4(8.51%),27-28 years S. aureus was 3(6.38%). The differences in presence of S. aureus (F.Cal=17.552) in relation to different age group was statistically significant. The colonization of S. aureus was in male 30(63.8%)(Table 4.4, figure 4.4) higher than in female 17(36.1%), which was closed to previous study (56.8%) in male and 43.2% in female. The differences in presence of S. aureus (F.Cal=29.138) in relation to different gender was statistically significant. All the isolated were resistance to erythromycin and penicillin 47 (100%). The S. aureus isolates showed high resistance to erythromycin, penicillin, Amoxicillin and clindamycin (100%, 100%, 97.87%, 91, 48%) respectively. Antibiotic resistance rates for gentamicin, vancomycin were moderate ( 53.19%, 78.72%) respectively, while S. aureus isolates showed low resistance against tetracycline, ciprofloxacin and trimethoprime (19.14%, 6.38% and 44.68%). The results of isolation of DNA from erythromycin resistance of Staphylococcus aureus were by PCR technique. Fourty seven (47) of S. aureus were resistance to erythromycin, these isolates contain any of erythromycin resistance genes tested (erm A, B, C). In our study, the most prevalent resistance gene in S. aureus was erm A (29/47; 61.7%), followed by erm C (11/47;23.40%). Less common were erm B which occurring in (7/47; 14,89%) of the erythromycin resistant S. aureus isolates tested. The differences in prevalence of erm A,B and C (p=7.815) in S. aureus was statistically significant. In this study 16SrRNA detected in 45 (95.74%/)out of 47 S. aureus samples.ThesisItem Open Access Characterization and praxis of keratinase produced from Bacillus spp.(Department of Industrial Microbiology Jacob Institute of Biotechnology and Bioengineering Sam Higginbottom University of Agriculture, Technology & Sciences Allahabad (U.P.) India, 2018) Singh, Sandeep; Masih, Dr. HarisonThe present study was mainly focused on the isolation and identification of keratinolytic isolate and their utilization in production and optimization of cultural conditions to maximize the keratinase production. Also focused for purification and characterization of produced keratinase. Some applications of keratinases were also studied. Out of 100 soil samples 25 keratinolytic bacterial strains were isolated and identified. Two selected isolates were identified as Bacillus subtilis S1 and Bacillus pumilis S2 according to 16 S rRNA sequencing. The influence of cultivation temperature and initial pH of the medium on keratinase production by Bacillus subtilis S1 and Bacillus pumilis S2 revealed the optimum values for the temperature and pH as (40°C and 7) and (40°C and 8), respectively. Maximum keratinolytic activity was observed at 72h and 96 h in S1 and S2, respectively after incubation. Optimized value for inoculum size and substrate concentration was found to be (5% and 1%) and (3% and 1%) in S1 and S2 , respectively, 150 rpm found to be the optimum agitation level in both the cases. The optimum nitrogen source was beef extract and peptone in S1 and S2, respectively and all the carbon sources showed negative effect on keratinase production by both isolates. DEAE-Cellulose chromatography resulted in purification yield of (5.5% and 4.8%) and a purification fold of (5.64 and 5.84) in S1 and S2, respectively. After characterization S1 and S2, keratinases found stable in a broad pH range 5-11 with optimum at pH 8 and pH 9. Both the keratinases from S1 and S2, found stable in a broad temperature range 30-70°C with optimum at 50°C and 40°C. Km and Vmax values for S1 were 0.128 mg/ml and 8 μm/ml/min, respectively. Km and Vmax values for S2 were 0.359 mg/ml and 11.24 μm/ml/min, respectively. Molecular weight of both the enzymes from S1 and S2 were 29.8 kDa and 35 kDa, respectively. Both the isolates showed remarkable dehairing of hides and washing effect. Both the isolates could be utilized as prospective microbes for profitable application in feather degradation, in dehairing in tanneries in place of harmful chemicals and could furthermore stimulate proficient solid waste management, where unremitting amassing of feather wastes causes severe environmental problems.ThesisItem Open Access DEVELOPMENT AND VALIDATION OF A MICROARRAY CHIP FOR DIAGNOSIS OF ANIMAL VIRUSES(DEPARTMENT OF BIOCHEMISTRY INDIAN VETERINARY RESEARCH INSTITUTE IZATNAGAR, 2016) Rai, Gaurava Kumar; Lawrence, RubinaThesisItem Open Access Bacterial isolates and drug susceptibility patterns of urinary tract infection among pregnant and non-pregnant women(FACULTY OF HEALTH SCIENCES SAM HIGGINBOTTOM INSTITUTE OF AGRICULTURE, TECHNOLOGY AND SCIENCES Deemed-to-be-University (FORMERLY ALLAHABAD AGRICULTURAL INSTITUTE), 2016) ABULMESHAH, MANSSUOR ABULKASIM AHMED; Shukla, Prof. (Dr.) G. S.; Bajaj, Prof. (Dr.) H. K.The study ʻʻBacterial isolates and drug susceptibility patterns of urinary tract infection among pregnant and non-pregnant womenʼʼ was carried in Sam Higginbottom Institute of Agriculture Technology and Sciences (Deemed to be University), Allahabad India during the period of June 2013 to October 2016. Five hundred (500) urine samples were collected and analyzed during the study period in BioAxis DNA Research Centre, L.B. Nagar Hyderabad India between March 2015 to August 2016. The aim of this study is to isolate and identify bacteria on the basis of their cultural, morphological & biochemical characteristics, also to identify the specific strains of isolated bacteria which cause UTI by PCR and 16s rRNA sequence technique, and to study the antibiotics resistance pattern of the isolated from urine sample of pregnant and non-pregnant woman. Out of five hundred (500) urine samples, one hundred seventy nine (179) samples showed significant growth, which amounted to a prevalence of 35.8%. The incidence of UTI in pregnant women (65.3%) was higher than the incidence of non-pregnant women (34.6%). Of the 179 isolates obtained, Gram negative rods 103 (57.54%) occurred more frequently than Gram positive cocci 76 (42.45) constituting the total isolates. Among the significant isolates, Staphylococci aureus had the highest percentage isolation with frequency rate 76 (42.45%) [94 (41.88%) in pregnant women and 27 (43.54%) in non-pregnant women], followed by Escherichia coli was 59 (32.96 %) [39 (33.33 %) in pregnant women and 20 (32.25 %) in non-pregnant women], Klebsiella spp was 23(12.84%) [14 (11.96%) in pregnant women and 9 (14.51%) in non-pregnant women], Pseudomonas areuginosa was 17(9.49%) [12 (10.25%) in pregnant women and 8 (8.06%) in non-pregnant women], while the lowest was Proteus spp 4(2.23%) [3 (2.56%) in pregnant women and 1 (1.61%) in non-pregnant women]. The prevalence rate of 117 pregnant patients in relation to the trimester of pregnancy was higher in the second trimester (52.13%) compared to the first (16.23%) and third (31.62%) trimesters. ii The BLAST analysis of the 16s rRNA region of the DNA sequences of the five bacterial isolates showed the following results, isolate (Bsequ-01) revealed 100% similarity to Escherichia coli and thus the isolate Bsequ-01 was confirmed as Escherichia coli. The bacterial isolate (Bsequ-02) revealed 96% similarity to Pseudomonas aeruginosa. The bacterial isolates (Bsequ-03, Bsequ-04 and Bsequ-05) revealed 100%, 97% and 100% similarity to Proteus mirabilis, Klebsiella pneumoniae and Staphylococci aureus respectively. Amikacin was found to be the most effective drug (97.2%) followed by Cefotaxime (86.5%). In this study there was no statistically significant value in antibiotic susceptibility patterns of uropathogen between pregnant and non-pregnant women due to P- value more than 0.05(P > 0.05). Key words: Urinary Tract Infection, UTI, Antibiotic Resistance, PCR, DNA SequencesThesisItem Open Access EVALUATION OF PERFORMANCE OF PLANT GROWTH PROMOTING BACTERIA (PGPB) CONTAINING 1-AMINOCYCLOPROPANE- 1-CARBOXYLATE DEAMINASE (ACCD) IN IMPROVING GROWTH AND YIELD OF CEREAL CROPS UNDER SALINITY STRESS(DEPARTMENT OF INDUSTRIAL MICROBIOLOGY JACOB INSTITUTE OF BIOTECHNOLOGY AND BIOENGINEERING SAM HIGGINBOTTOM UNIVERSITY OF AGRICULTURE, TECHNOLOGY AND SCIENCES ALLAHABAD 2017, 2017) SAGAR, ALKA; Ramteke, Pramod W.Soil salinity is an important problem for crop production in many parts of the world, especially in irrigated fields of arid and semiarid regions. Plant growth promoting bacteria (PGPB) are known to reduce adverse effect of environmental stress and thereby enhance the growth and development of plant health, soil fertility and agricultural sustainability. PGPB contain enzyme 1-aminocyclopropane-1-carboxylate deaminase (ACCD) that inhibits the ethylene production from its precursor ACC into α-ketobutyrate and ammonia. Removal of ACC significantly increases plant root and shoot length and protect the plant from stress. In the present study, 650 bacterial cultures were isolated from the soil of Model Organic Farm (SMOF) of Sam Higginbottom University of Agriculture, Technology and Sciences (SHUATS), Allahabad, India. Rich bacterial diversity in soil from SMOF both in terms of their types and functional plant growth promoting (PGP) traits was observed and represented by heterotrophs, coliforms, Pseudomonas spp., Azotobacter spp. and Rhizobium spp. and majority of them displayed multiple plant growth promoting (MPGP) traits. Bacterial isolates were predominately positive to production of ammonia (NH3) (93.2%), indole acetic acid (IAA) (89.6%), catalase (85.0%), 1-aminocyclopropane- 1-carboxylate deaminase (ACCD) (78.6%) and siderophore (69.0%). Richness of their functional characteristics is further revealed by their tolerance to salinity and wide range of pH. In the present study all isolates from organic farm were tolerant to > 5 % NaCl and wide range of pH. We obtained major (96-97%) constituent of bacterial population of nitrogen fixers Azotobacter spp. and Rhizobium spp. with multiple PGP traits and tolerance to salinity and wide range of pH. Twenty nine (29) potential PGPB were identified on the basis of molecular with 16S rRNA and sequences were submitted to NCBI and got different accession no. Also the potential PGPB were tolerant to high levels of trace elements, and resistant to multiple antibiotics. On the basis of tolerance to high salt (20%) concentration XVIII three salt tolerant PGPB (STPGPB) viz. E. cloacae (KP226569) (PR4), Enterobacter sp. (KP226570) (PR14) and A. nigricans (KP966496) (PR19) were selected for detailed studies. All the three STPGPB expressed significant increase (p<0.001) antioxidant enzymes activities (SOD, CAT and GSH) under the abiotic stress of salinity and pH. Inoculation with STPGPB exhibited higher percentage of seed germination and enhanced growth parameters in cereals seed such as rice, maize and millets under salinity stress. Further significantly (p<0.001) increased percentage of seed germination and enhanced growth of these cereals was noted under salinity in presence of ammonium sulfate (substitute of ACC) clearly demonstrating the role of ACCD in alleviation of abiotic stress, especially salinity.ThesisItem Open Access Genomic diversity of antagonistic Trichoderma species against phytopatogen of Lycopersicon Esculentum Mill(Dept. of Microbiology, Sam higginbottom university of agriculture , technology and science, 2017) Rai, ShaliniThesisItem Open Access Salt induced alterations in the Physiological, Biochemical and Proteomic profile of the symbiotic cyanobacterium Anabaena azollae(Sam Higginbottom Institute of Agriculture, Technology & Sciences (SHIATS), 2016) Yadav, Ravindra Kr.; W. Ramteke, PramodThe aquatic nitrogen fixing pteridophyte Azolla is an excellent biofertilizer for rice paddy fields owing to its ability fix to atmospheric nitrogen. However, wider exploitation of Azolla is hampered due to increasing soil salinity. Understanding the physiological response of the cyanobiont to salinity is therefore important to unravel the salinity tolerance mechanism operating in this system. This will further pave way for the intervention of advanced molecular biological tools to work out the mechanisms operating under salinity stress conditions in the cyanobionts. Therefore in the present study the physiological and biochemical response of freshly separated cyanobionts from two different species of Azolla viz. A. microphylla and A. caroliniana exposed to salinity was studied. Freshly separated cyanobionts of Azolla microphylla and Azolla caroliniana plants exposed to salinity showed decline in the cellular constituents such as chlorophyll (23.1 and 38.9%) and protein (12.9 and 19.3%). However, an increase in the carotenoid and sugar content was observed. Exposure to salinity stress reduced the heterocyst frequency (35.4 and 57.2%) and nitrogenase activity (37.7 and 46.3%) of the cyanobionts. Increase in the activity of antioxidant enzymes such as super oxide dismutase (50.6 and 11.5%), ascorbate peroxidase (63.7 and 57.9%), catalase (94.2 and 22.5%) as well as non-enzymatic antioxidant proline (18.8 and 13.3 %) was also observed in response to salinity. The cyanobionts exhibited significant increase in the intracellular Na+ level and reduced intracellular K+/Na+ and Ca2+/Na+ ratio in response to salinity. The results demonstrated the adverse impact of salinity on the freshly separated cyanobionts as similar to free living cyanobacteria. Salinity stress response mechanisms in the freshly isolated cyanobionts Anabaena azollae was studied at the proteome level using two dimensional gel electrophoresis (2DE) followed by MALDI-TOF-MS/MS analysis. Thirty five (35) protein spots were significantly altered due to exposure to salinity. Among them, thirteen (13) spots were down regulated and twenty two (22) spots were up regulated. A total of twenty (20) new or hypothetical proteins have also been identified in the present study. These proteins are associated with a variety of functions including energy metabolism protein synthesis and folding, DNA damage and repair, stress and defense. The study might help in understanding the biological processes and stress proteins involved in salinity stress adaptation. Results show that the cyanobiont resorts to elaborate changes in the protein synthesis and cell signaling to survive under salinity induced stress conditions. These results may be helpful in the critical evaluation of salinity tolerance mechanism of the cyanobiont and its interaction with the host.