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Anand Agricultural University, Anand
Anand Agricultural University (AAU) was established in 2004 at Anand with the support of the Government of Gujarat, Act No.(Guj 5 of 2004) dated April 29, 2004. Caved out of the erstwhile Gujarat Agricultural University (GAU), the dream institution of Sardar Vallabhbhai Patel and Dr. K. M. Munshi, the AAU was set up to provide support to the farming community in three facets namely education, research and extension activities in Agriculture, Horticulture Engineering, product Processing and Home Science. At present there seven Colleges, seventeen Research Centers and six Extension Education Institute working in nine districts of Gujarat namely Ahmedabad, Anand, Dahod, Kheda, Panchmahal, Vadodara, Mahisagar, Botad and Chhotaudepur
AAU's activities have expanded to span newer commodity sectors such as soil health card, bio-diesel, medicinal plants apart from the mandatory ones like rice, maize, tobacco, vegetable crops, fruit crops, forage crops, animal breeding, nutrition and dairy products etc. the core of AAU's operating philosophy however, continues to create the partnership between the rural people and committed academic as the basic for sustainable rural development. In pursuing its various programmes AAU's overall mission is to promote sustainable growth and economic independence in rural society. AAU aims to do this through education, research and extension education. Thus, AAU works towards the empowerment of the farmers.
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ThesisItem Open Access PHENOTYPIC AND GENOTYPIC IDENTIFICATION AND METAGENOMIC ANALYSIS OF SUBCLINICAL MASTITIC PATHOGENS IN COWS(AAU, Anand, 2011) BHANDERI, BHARAT BABUBHAI; Jhala, M. K.Subclinical mastitis occurs with no visible changes in the appearance of the milk and/or the udder, but milk production decreases which leads to economic losses to the farmers and dairy industry. There are many microbial pathogens involved in causing subclinical mastitis in cows. The present study was undertaken to know incidences of subclinical mastitis in organized farms using Somatic Cell Count (SCC) and bacteriological examination (International Dairy Federation-IDF guidelines), California Mastitis Test (CMT) and impregnated pH strip test followed by characterization and PCR based detection of important mastitic pathogens. Metagenomic analysis of subclinical mastitis milk was also done to determine the complex microbial diversity in udder environment during subclinical mastitis. A total of 349 quarters of 89 lactating cows comprising 31 Triple cross (TP) (Kankrej x Jersey x Holstein Friesian), 29 Kankrej, 17 Gir and 12 Holstein Friesian (HF) affiliated with Anand Agricultural University, Anand were screened for subclinical mastitis. Overall 52.8 per cent (47/89) cows were found to be positive for subclinical mastitis infection in one or more quarters. The highest incidence of subclinical mastitis was found in Triple cross cows (74.19%), followed by Gir cows (58.82%), HF cows (50%) and Kankrej cows (27.58%). Overall quarter wise incidence for subclinical mastitis was found to be 30.66 per cent (107/349). The highest incidence was found in Gir cows (38.80%) followed by Triple cross cows (38.08), HF cows (33.33%) and Kankrej cows (15.04%). The highest incidence of subclinical mastitis was found in fore left quarter (28.03%), followed by hind left quarter (27.1%), fore right quarter (24.29%) and hind right quarter (20.56%). Of the 47/107 cows/quarters positive for subclinical mastitis, 39/47 (82.97%) cows and 82/107 (76.63%) quarters were also positive by CMT and 27/47 (57.44%) cows and 56/107 (52.33%) quarters were positive by impregnated pH strip test. Cultural isolation ft'om 107 subclinically positive quarter milk samples yielded 126 bacterial isolates. Staphylococci was the most predominant bacterial species accounting for 53.97 per cent (68/126) of all the isolates, followed by 21.43 per cent (27/126) CAMP (Christie-Atkins-Munch-Peterson) test positive Str. agalactiae, 18.25 per cent (23/126) Micrococci, 4.77 per cent (6/126) E. coli and 1.58 per cent (2/126) Bacillus species. Out of 68 Staphylococci isolates, 38 (55.89%) isolates showed fermentation on Mannitol Salt Agar (MSA), whereas 30 (44.11%) isolates were mannitol non fermentive. Of the total 30 S. aureus identified by PCR, 21 (70%) were mannitol fermentive and 9 (30%) mannitol non fermentive. Thirty one (45.58%)) Staphylococci were found to be positive for pigment production, whereas 37 (54.42%) isolates produced white colonies on nutrient agar. Forty eight (70.58%) isolates were found positive for coagulase reaction, whereas 20 (29.41%) were negative. Thirty one (45.58%)) isolates exhibited P haemolysin production, 4 (5.89%) a haemolysin and 33 (48.53%)) isolates were non-haemolytic on 5 per cent Sheep blood agar. Phage typing at National Staphylococci Phage typing Centre, Maulana Azad Medical College, New Delhi, using five phage group sets of International Basic Set of 23 phages revealed maximum number of the Staphylococci isolates lysed by group II 14 (82.35%), followed by groups III, Not alloted (NA), I and V with 12 (70.58%), 9 (52.94%), 5 (29.23%) and, 2 (11.76%) respectively. Maximum 11 (64.7%) isolates were lysed with phage number 47 with strong reaction, followed by 10 (58.82%)) isolates with phage numbers 42E and 81, while less effective phage numbers were 71 and 94, which lysed only one strain (5.89% each) and phage number 95 not giving strong reaction with any of the isolates. The methicillin and oxacillin antibiotic sensitivity pattern by disc diffusion method revealed that, all the 68 (100%)) Staphylococci isolates were sensitive. Serotyping of six E. coli isolates (at National Salmonella and Escherichia Centre, Kasauli, Himachal Pradesh for 'O' antigen) resulted in identifying 014, O20, 045, 055 and 0112 serotypes, while one isolate was untypeable (UT). Out of 68 Staphylococci isolates tested for identification of 5. aureus by PCR, 30 isolates were identified as S. aureus by obtaining amplification product of 1318bp using S. aureus specific primer for 23S rRNA. Out of 30 PCR positive S. aureus, 18 (60%)) were positive and rest were negative for coagulase test. All the 27 Streptococci isolates were identified as Str. agalactiae by amplifying 586bp product using Str. agalactiae specific primer for the 16S rRNA while, none were amplified for Str. dysgalactiae (401bp) and Str. uteris (94bp) based on primers specific for the 16S rRNA and 23 S rRNA respectively. All the six E. coli isolates yielded 232bp amplified product using E. coli specific primer targeting DNA sequence coding for the 23 S rRNA. Metagenomic analysis (using GS FLX 454 Life Sciences) of DNA of subclinical mastitis milk sample of TP, Kankrej and Gir cows yielded an out put of 274190 bp, 17,727 bp, 42,548 bp and 1,960, 170, 301 contigs respectively. Average fragment length obtained were 139.89, 104.28 and 141.36 bp for TP, Kankrej and Gir cows respectively. The longest sequence length was 560, 327 and 454 bp, while shortest sequence length was 40, 40, and 41 bp for TP, Kankrej and Gir cows respectively. A total of 54 (2.76%), 39 (22.94%) and 12 (3.99%) sequences for TP, Kankrej and Gir cows respectively could be matched to proteins in SEED subsystems of MG-RAST (Meta Genome Rapid Annotation with Subsystem Technology) (using an e-value cut-off of le-5). Metagenomic analysis of the three breeds identified bacterial organisms belonging to phyla (5), class (8), Subclass / order (15), Family (19), Genus (23) and species (28); of these, 19 genera and 26 species, many of which were fastidious/anaerobic organisms, were identified additionally than the cultural methods. Out of five genera Staphylococcus, Streptococcus, Micrococcus, Bacillus and Escherichia detected in the subclinical mastitis milk samples of TP, Gir and Kankrej breeds by culture based methods, four genera Staphylococcus, Streptococcus, Bacillus and Escherichia were also identified in the corresponding pyrosequencing data, while Micrococcus identified by culture based methods was not found in the pyrosequencing data. In pyrosequencing, over all 28 bacterial species were identified from all the three breeds of cows viz. Leifsonia xyli, Propionibacterium acnes, Streptomyces coelicolor, Chlamydophila abortus, Staphylococcus aureus, Staphylococcus epidermidis, Lactobacillus acidophilus, Streptococcus mitis, Burkholderia cenocepacia, Burkholderia cepacia, Ralstonia solanacearum, Nitrosomonas europaea, Pseudoalteromonas atlantica. Salmonella Dublin, Serratia marcescens, Azotobacter vinelandii, Pseudomonas aeruginosa, Pseudomonas mendocina, Stenotrophomonas maltophilia. Bacillus subtilis, Lactobacillus delbrueckii. Aster yellows witches'-broom phytoplasma, Pannbaculum lavamentivorans, Thermosipho melanesiensis, Aeromonas hydrophila, Escherichia coli, Shigella hoydii and Pseudomonas fluorescens. Of these, except S. aureus and E. coli, all were additionally identified than the culture based method but, Str. agalactiae identified by cultural method was not found in the pyrosequencing data. The role of lesser known or less frequently involved organisms as identified by metagenomic analysis may be further explored in future so as to understand the complete etiopathology of subclinical mastitis in cows.ThesisItem Open Access LABORATORY STUDIES ON BIOLOGY AND FEEDING POTENTIAL OF MEXICAN BEETLE, Zygogramma bicolorata Pallister ON PARTHENIUM, Parthenium hysterophorous L. AND EVALUATION OF HERBICIDES FOR THEIR SAFETY TO THE BIOAGENT(AAU, Anand, 2013) PAWAR, SATISH RAMCHANDRA; Korat, D. M.Investigations on biology, morphometries and feeding potential of Mexican beetle, Zygogramma bicolorata Pallister (Chrysomelidae: Coleoptera) and toxicity of weedicides as well as effect of temperature on biological attributes of the bioagent were carried out in Biological Control Research Laboratory, Anand Agricultural University, Anand (Gujarat) during theyear 2011 to 2012. Studies on biology of Z. bicolorata on Parthenium hysterophoms L. revea:ied that the female laid their eggs either singly or in cluster on under surface of leaves. Eggs were oblong in shape, slightly elongated, smooth and the surface was finely reticulated. There were four distinct larval instars. Newly hatched larvae were yellowish in colour and gradually turned in creamy white with the advancement of age. Adults of Z. bicolorata were elongate and oblong in shape. Dorsal surface was strongly convex and glabrous. In general, females found relatively larger in size than males. Average egg, larval, pupal and adult period was 3.93+0.80, 13.87 ± 1.36, 7.40 ± 1 . 1 8 and 32.40 ± 8.05 (males) to 44.53 + 7.33 (females) days, respectively. Entire life-span of female and male completed in 59.13 ± 7.75 and 71.33 ± 8.78 days, respectively. Fecundity, hatching percentage, adult emergence percentage and male to female sex ratio was 669.73 ± 141.34 eggs, 55.75 ± 12.27%, 76.67 ± 14.82% and 1 : 1.26, respectively.ThesisItem Open Access INFLUENCE OF DECAPITATION AND PGR's ON GROWTH, YIELD AND QUALITY OF CLUSTER BEAN SEED (Cyamopsis tetragonoloba L. Taub.) CV. PUSA NAVBAHAR(AAU, Anand, 2010) NARSHI, SATODIYA BAKUL; Patel, H. C.A field experiment was conducted at Main Vegetable Research Station, Anand Agricultural University, Anand during the years 2008-09 and 2009-10 in summer seasons on sandy loam soil with a view to study the "Influence of decapitation and PGR's on growth, yield and quality of cluster bean seed (Cyamopsis tetragonoloba L. Taub.) cv. Pusa Navbahar" under middle Gujarat conditions. There were twenty one treatment combinations comprising three decapitation treatments i.e. D1-Without decapitation (control), D2-Decapitation at 70 DAS and Ds-Decapitation at 85 DAS and seven different PGR's treatments along with control i.e. G1 -Water spray (Control), G2-NAA 20 mg/l, G3-NAA 40 mg/l, G4-GA3 20 mg/l, G5-GA3 40 mg/l, G6-Thiourea 500 mg/l and G7-Thiourea 1000 mg/l in a Randomized Block Design (Factorial) with three replications. Growth parameters viz. plant height and number of leaves per plant were found non-significant at 30 and 60 DAS. Whereas, without decapitation (control) treatment recorded significantly the highest plant height i.e. 112.07 and 115.72 cm and number of leaves per plant i.e. 50.70 and 45.15 at 90 DAS and harvest stage, respectively. The effects of decapitation on physiological parameters viz. LA, LAI, CGR, NAR and dry weight of plant were found non-significant at 30 and 60 DAS whereas, at 90 DAS all the physiological parameters and at harvest only LA and dry weight of plant recorded significantly, higher values in control treatment (Without decapitation). Decapitation of plant at 70 DAS registered significantly the highest number of pods per cluster (7.55), number of clusters per plant (18.29), number of pods per plant (138.00) and weight of 1000 seeds (37.00 g) in pooled analysis. However, dry pod length (9.59 cm) and number of seeds per pod (7.05 cm) were found maximum in treatment decapitation at 85 DAS. Decapitation of plant at 70 DAS produced significantly the highest seed yield (917.7 kg/ha) in pooled analysis as compared to without decapitation (889.13 kg/ha), which was 9.26 % higher than without decapitation. Root and shoot ratio (0.057) and harvest index (627.04) were also found significantly superior in treatment decapitation at 70 DAS. Seed quality parameters viz. seedling length, seedling dry weight, vigour index-I and II were recorded the highest values in treatment decapitation at 70 DAS at 6th 8th and at 10th day on pooled basis. Plant growth regulators treatments significantly influenced growth parameters viz. plant height and number of leaves per plant at various growth stages of crop except at 30 DAS. At 60, 90 DAS and harvest stage treatment GA3 40 mg/l recorded significantly maximum plant height and number of leaves per plant as compared to rest of treatments. Physiological parameters viz. LA, LAI, NAR and dry weight of plant at 60 and 90 DAS registered significantly the higher values in treatment GA3 40 mg/l in pooled analysis. At harvest, LA, LAI and dry weight had recorded significantly maximum values in treatments Thiourea 500 mg/l and GA3 40 mg/l which remained at par with each other. Yield attributing characters viz. number of pods per cluster (7.25), number of clusters per plant (18.75), number of pods per plant (136.01), dry pod length (9.86 cm) and weight of 1000 seeds (37.22 g) were significantly the highest in treatment Thiourea 500 mg/l on pooled basis. The significantly maximum seed yield was recorded by the treatment Thiourea 500 mg/l (1030.36 kg/ha) in pooled analysis, which was at par with treatment GA3 20 mg/l (992.81 kg/ha) which was 26.67 and 22.05 %, higher than control, respectively. The treatment GA3 20 mg/l recorded the highest harvest index (638.80). Seed quality parameters viz. germination percentage, seedling length, seedling dry weight and vigour index-I and II were recorded the maximum values in treatments of GA3 and Thiourea at all the three stages i.e. 6th, 8th and 10thday. Interaction effects between decapitation and PGR's treatments on growth parameters viz., plant height and number of leaves per plant showed significant differences at 60, 90 DAS and harvest stage except for number of leaves per plant at harvest. Different combination of decapitation and PGR's treatments recorded significant results on physiological parameters at various growth stages viz. LA, LAI, CGR, NAR and dry weight of plant. At 60 and 90 DAS LA, LAI, NAR and dry weight of plant recorded significant results in different combinations except CGR at 60 DAS. At harvest stage, CGR, NAR and dry weight of plant were found significant due to interaction between decapitation and PGR's treatments. At all the growth stages, interaction between without decapitation and treatments of GA3 and Thiourea recorded significantly higher values of all the physiological parameters. Yield attributing parameters viz. number of seeds per pod and weight of 1000 seeds significantly affected by interaction between decapitation and PGR's treatments which, were the highest in combinations D'aGe (8.20) and D2G6 (41.40 g), respectively. Seed quality parameters like, seedling length, seedling dry weight and vigour index-I and II recorded the maximum values in combinations of decapitation at 70 DAS with treatments of GA3 and Thiourea at all the three stagesi.e. 6th 8th and10th day. The higher net return of 47,849 Rs./ha along with B.C.R. of 1: 2.60 was obtained by decapitation at 70 DAS, whereas, the lowest net realization (41,935 Rs. /ha) and B.C.R. (1:2.43) was obtained when plant decapited at 85 DAS. The treatment Thiourea 500 mg/l recorded the maximum net realization of Rs. 52,277 per hectare with 1:2.73 B.C.R., followed by treatment GA3 20 mg/l.ThesisItem Open Access EXPRESSION PROFILING, SNP DETECTION AND VALIDATION IN SQUAMOUS CELL CARCINOMA OF HORN IN KANKREJ CATTLE (Bos indicus) USING NEXT GENERATION SEQUENCING(AAU, Anand, 2014) KORINGA, PRAKASHKUMAR G.; Joshi, Chaitanya G.Horn cancer is a widely prevalent cancer amongst Kankrej cattle (Bos indicus) seen sporadically, especially in case of working class of castrated male animals i.e. bullocks. A transcriptome envisaged characterization as well as correlation to known genomic changes such as structural and copy number alterations, focused ins/dels and single nucleotide mutations. Here, we employed high throughput RNA-seq using GS-FLX Titanium for characterization and comparison of normal and cancerous horn transcriptome in Bos indicus. A total of 909,362 reads with average read length of 405bp for horn cancer (HC) and 583,491 reads with average read length of 411bp for horn normal (HN) were obtained by sequencing gene transcripts derived from HC and HN tissues. Assembled data were analyzed for identifying novel as well as differentially expressed transcripts using CLC Genome Workbench. RNA-seq analysis using different bioinformatics pipelines and software identified differentially expressed genes i.e. upregulation of KRT6A, KRT6B, KRT6C, KRT14, SFN, KRT84, PI3, CAl, C0L17A1, ANLN, SERPINB5 etc., as well as down-regulation of NR4A1, FOSB, LRIGl, BOLA, SCGBIAI, CXCL17, KRT19, BPIFBl, NR4A1 and TFF3 etc., in HC tissues. The signaling pathway investigation in this study revealed many of the cancer related pathways which mainly include cell cycle regulation pathways, p53 tumor suppressor pathways, NFKB and MAPKs pathways, LPS signaling pathway and PI3K-Akt pathways. The resuh of transcriptome expression profiling was validated using RT-qPCR in nine randomly selected genes. It revealed concordance of gene expression profile with RNA-seq analysis. We also used transcriptome data to elucidate complexity of the alternative splicing in HC transcriptome. We identified potential candidate splice variants that might be helpful in development of relevant biomarkers for early diagnosis of HC. The fiiture studies targeted at in depth characterization of these potential candidate splice variants might change the currently used clinical approaches. Herein we characterized global landscape of alternative splicing events exhibited by pair of HC and HN tissue and confirmed selected alternative splicing events with significant association to HC by RT-qPCR. Ine analysis of the same RNA-seq data using SeqMan Pro Version 10.0.0 resulted in to a 9532 and 7065 SNPs as well as 1171 and 1172 Indels in HC and HN, respectively. Out of total, 7889 SNPs and 1736 Indels uniquely present in HC, 5886 SNPs and 1146 Indels uniquely present in HN are novel and reported first time in Bos indicus, whereas rest are already reported in Bos taurus dbSNP database at NCBI. The gene-associated SNPs and Indels were high in upregulated genes of HC as compared to HN tissues. SNPs identified in RNA-seq analysis were validated in fiirther studies in two groups consisting of 50 animals each of HC and HN bullocks. DNA from HC tissue and blood of HN individual was extracted and 96 pairs of primers were used to generate amplicons of an average 300bp to get sequenced using Ion Torrent PGM. The resulting reads were assembled using SeqMan N Gen of DNASTAR and data were analyzed using Arraystarll. Case control analysis was carried out to find SNP significantly associated with HC. SNP at position 63251805 (dBSNP ID rsl36870681) identified in BPIFAl can serve as a potential candidate genetic marker in HC. The SNPs and Indels identified in this study will be useful resource for future studies to understand genetic basis for phenotypic variation between Bos taurus and Bos indicus as well as cancers in animals. A very large number of SNPs are essential for the designing and construction of arrays. SNPs identified in this study will enrich the dbSNP database of NCBI (http://www.ncbi.nlm. nih.gov/projects/SNP/) and will be useful resource for array designing. This study is the first attempt to reveal novel transcripts, differentially expressed genes as well as identification and validation of SNPs using digital expression analysis in Bos indicus and provides novel insights into bovine transcriptome. Our study will serve as a step further in detailed characterization of HC transcriptome and provide firm base to explore and mitigate HC at finer resolution. The present findings would provide basis for further screening of genes and identification of markers for early diagnosis and therapeutic intervention of HC.ThesisItem Open Access HAEMATOLOGICAL, BIOCHEMICAL AND ENDOCRINE PARAMETERS AT DIFFERENT AGES AND PHYSIOLOGICAL STAGES IN GIR CATTLE AND JAFFARABADI BUFFALOES(AAU, Anand, 2012) JACOB, NINAN; ARYA, J. S.The study titled "Haematological, Biochemical and Endocrine parameters at different Ages and Physiological stages in Gir cattle and Jaffarabadi buffaloes" was imdertaken in different age groups and physiological stages in Gir cattle and Jaffarabadi buffaloes with the objective to determine and compare the species differences for (i) haematological parameters viz. TEC, Hb, PCV, TLC, DLC (ii) biochemical parameters viz. plasma glucose, total protein, albumin, total cholesterol, HDL cholesterol, triglycerides, urea, uric acid, creatinine, calcium, phosphorus, magnesium, sodium and potassium (iii) enzymes viz. aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, acid phosphatase, and lactate dehydrogenase (iv) hormonal levels of triiodothyronine, thyroxine, insulin, growth hormone, Cortisol, estradiol - 17 β, progesterone and testosterone and (v) to observe the milk components viz. percentage of fat, protein and lactose in lactating Gir cows and Jaffarabadi buffaloes and their relationship with the stage of lactation under study. The blood samples were collected from Gir and Jaffarabadi females (n=8 for each sampling stage) at 1 wk, 1, 3, 6, 12, 24 and 36 months age, at 1, 2 and 3 month of lactation and in non-lactating pregnant and non-pregnant animals. In Gir and Jaffarabadi males (n=6 for each sampling' stage) blood samples were collected at 1 wk, 1, 3, 6 and 12 month of age and in bulls. Castrated males were also used for blood sampling in Gir males. A total of 270 blood samples were collected and the analysis was conducted by standard techniques.ThesisItem Open Access MONITORING POSTPARTUM REPRODUCTIVE PERFORMANCE IN KANKREJ COWS THROUGH CLINICAL DIAGNOSIS, BLOOD PROFILE AND HORMONAL THERAPY(AAU, Anand, 2012) NAIKOO, MEHRAJUDDIN; Dhami, A. J.The present study was carried out at Livestock Research Station of the University on 42 Kankrej cows of 2nd to 4th parity. The chief objectives were: to monitor the early postpartum period (0-90 days) clinically and through plasma profile of progesterone, metabolites and macro-micro minerals at 10 days intervals; to evaluate the efficacy of a sustained release mineral supplement (Mega bolus PO) on the day of calving and five oestrus induction and synchronization protocols (Ovsynch, CIDR, Ovsynch + CIDR, Cosynch and PGF2α) on day 90-95 postpartum towards augmenting reproductive efficiency of anestrous and subestrous cows (6 animals in each group), keeping 6 normal cyclic animals as control, and its effect on above profile till day 40 post-treatment/post- AI, and to compare plasma profiles of conceived and non-conceived cows at first Al. The time required for expulsion of fetal membranes, weight of expelled fetal membranes and the birth weight of calf (pure and crossbred) were 5.04 ± 2.0 hrs, 2.84 ± 0.76 kg and 24.29 ± 1.54 kg, respectively. The Kankrej cows showed complete uterine involution by mean interval of 36.80 ±1.21 (range 32-45) days postpartum. The interval for occurrence of first oestrus postpartum clinically and through plasma P4 profile was 105.49 ± 1.66 (range 86-106) and 56.42 ± 3.88 (range 30-80) days, respectively (P<0.05). The first service and overall conception rates obtained at spontaneous/ induced oestrus, within 150 days postpartum were 30.95 (13/42) and 40.47 (17/42) per cent. The comparative evaluation of the efficacy of five oestrus induction/ synchronization protocols tested, on 6 cows each, viz. Ovsynch, CIDR, Ovsynch + CIDR, Cosynch and PGF2α revealed oestrus induction response of 66.66, 83.33, 50.00, 66.66 and 66.66 per cent, respectively, with behavioural signs at FTAI as confirmed by palpation per rectum. The first service conception rates obtained were 16.66, 33.33, 16.66, 50.00 and 50.00 per cent, respectively, as compared to 33.33 per cent in normal cyclic control cows. The corresponding second service conception rates were nil, 25.00, 20.00, nil, nil and nil per cent, as compared to 25.00 per cent in untreated control animals. The overall conception rates of three cycles over the 45 days period were 33.33, 50.00, 33.33, 50.00 and 50.00 per cent, respectively, as against 50.00 per cent in normal cyclic group. The results of CIDR, Cosynch and PGF2α protocols were better than the Ovsynch and normal control groups.ThesisItem Open Access EFFECT OF PROGESTERONE AND HEAT STRESS ON REPRODUCTIVE PERFORMANCE OF DAIRY COWS AND VALIDATION OF CONTINUOUS BODY TEMPERATURE MEASUREMENT IN IDENTIFYING ESTRUS(AAU, Anand, 2012) SUTHAR, VISHAL S.; Dhami, A. J.The present study with four experiments and manifold objectives were conducted on German (HF) dairy cows at or affiliated farm facilities of Clinic for Animal Reproduction, Freie Universitaet Berlin, Germany, during January 2010 to June 2012. The overall objectives of this study were to evaluate: 1) performance of temperature data logger in in vitro and in vivo conditions, 2) validity of BT to identify induced estrus, 3) effect of exogenous and endogenous P4 on BT of dairy cows and 4) and to study effect of heat stress on BT and postpartum performance of dairy cows. In first study to evaluate performance of temperature data logger (Minilog 8, Vemco Ltd., Halifax, Canada), three sub-experiments were conducted. The study began with an in vitro validation of 24 temperature loggers comparing them to a calibrated liquid-in-glass thermometer as a reference method (sub-experiment 1). The association and agreement between the 24 temperature loggers with the reference method was r = 0.996 (P < 0.001) and a negligible coefficient of variance (0.005) between the loggers. In vivo temperature loggers were tested in 11 healthy postpartum cows (sub-experiment 2) and 12 early postpartum cows with greater BT (sub-experiment 3). Temperature loggers were set to record VT and RT at 1 min intervals. To prevent rectal and vaginal straining and potential expulsion of temperature logger an epidural injection of 2.5 ml of 2% Procain was administered. Association between RT and VT was r = 0.92 (P < 0.001) in sub-experiment 2 and r = 0.94 (P < 0.001) in sub-experiment 3 with a negligible difference of -0.1 and 0.01°C, respectively. Bland-Altman plots demonstrated an agreement between RT and VT for healthy and early postpartum cows with greater BT in sub-experiment 2 and 3, respectively. Therefore, continuous VT monitoring with temperature loggers can be used as a measure for BT in dairy cows.ThesisItem Open Access REGULATION OF ACTIVIN TYPE II RECEPTOR B (ACVR2B) EXPRESSION THROUGH RNA INTERFERENCE IN GOAT MYOBLAST CELLS(AAU, Anand, 2014) PATEL, AMRUTLAL KALUBHAI; Joshi, Chaitanya G.Enhancement of skeletal muscle mass through genetic manipulation has drawn attention to increase the meat production in the farm animals. Among the various techniques of regulating gene expression, RNA interference (RNAi) has been proposed as a promising tool to suppress the target gene expression. Attempts have been made to increase the skeletal muscle mass in transgenic animals through knockdown of Myostatin, a gene with potential negative effect on muscle growth. It has been well established that myostatin mediates its action through binding to its cell surface receptor mainly to activin type II receptor B (ACVR2B). Besides regulating myostatin activity, ACVR2B has also been known to regulate the activity of other Transfonning Growth Factor beta (TGF-(3) superfamily ligands which negatively regulates muscle growth. The inhibition of ACVR2B signaling has shown dramatic increase in the muscle mass to a greater extent than myostatin inhibition. Hence, in the present study we aimed to investigate the possibility of ACVR2B knockdown to enhance the myogenesis in goat through various RNAi methods such as expression of short hairpin RNAs (shRNAs) under U6 and CMV promoters and expression of artificial microRNAs (amiRNAs) under CMV and muscle specific muscle creatine kinase (MCK) promoters. Further we studied effect of ACVR2B knockdown on the expression of myogenic regulators and assessed induction of undesired interferon response against RNAi vectors. Among the seven shRNAs tested, the U6 promoter driven shRNAs showed 63 (p= 0.0004), 76 (p= 0.0001), 75 (p=0.0000), 74 (p= 0.0005), 80 (p= 0.0001), 74 (p= 0.0000) and 57% (p= 0.0013) silencing with sh1 to 7, respectively in HEK293T cells whereas 24 (p= 0.1497), 24 (p= 0.2243), 15 (p= 0.3988), 31 (p= 0.1263), 14 (p= 0.4425), 46 (p= 0.0318), and 26 % (p= 0.1288) silencing of endogenous ACVR2B with shl to sh7, respectively and 53 (p- 0.0005), 32 (p= 0.0171), 38 (p= 0.0025), 66 (p= 0.0002) and 51% (p= 0.0008) with sh3, sh4, sh5, sh6 and sh7, respectively against ectopically expressed goat ACVR2B in goat myoblasts ceils. The knockdown of endogenous ACVR2B resulted in the 116, 105, 84, 64, 119, 102, 121. and 157% expression of MyoD; 131, 128, 128, 123, 104, 103, 69, and 157% of MyoG in sh1 to sh7 transfected cells, respectively. The transfection of U6 driven shRNA resulted in the induction of OAS1, a marker for innate interferon response by 3 to 1861 fold in 293T cells and up to 94 fold in the goat myoblasts cells. In an attempt to overcome the undesired cellular toxicity associated with U6 driven shRNAs as reported in the number of studies, we expressed these shRNAs under CMV promoter. The CMV driven shRNAs showed weak silencing of 37 (p= 0.1622) and 18% (p= 0.4877) by sh1 and sh3, respectively in HEK293T cells whereas 7% (p= 0.5749) by shl and 4% (p= 0.7493) by sh5 in goat myoblasts cells. Unlike suggested earlier, we observed significant induction of interferon response to CMV driven shRNAs up to 46 fold in 293T cells and 105.3 fold in goat myoblasts cells. Alternatively, we assessed another RNAi approach using amiRNAs which mimics the endogenous miRNA biogenesis pathway. Among the four amiRNAs tested by placing them in 5'-UTR region of GFP reporter, we observed 64 (p=0.0004), 77 (p=0.0002), 1 (p=0.8712), and 41% (p=0.0115) silencing in 293T cells by ami204, ami318, ami735 and ami878, respectively against exogenously expressed goat ACVR2B; 19 (p=0.3593) and 9% (p=0.4977) by ami204 and ami318, respectively against endogenous ACVR2B and 23% (p=0.0444) by ami318 against exogenously expressed ACVR2B in goat myoblasts cells. Since, amiRNAs placed in 5'-UTR were shown to affect the translation of reporter GFP, we further placed them in 3'-UTR of GFP which resulted in enhanced expression of GFP thereby enabling the monitoring of expression of amiRNAs. The 3'-UTR derived amiRNAs showed 50% (p=0.0002) silencing only by ami3I8 in 293T cells whereas 47 (p=0.0193), 16 (p=0.2959), 19 (p=0.1547), and 28% (p=0.0770) by ami204, ami318, ami735 and ami878, respectively against endogenous and 67%) (p=0.0004) by ami318 against overexpressed ACVR2B in goat myoblasts cells. The expression of myogenic regulators MyoD remained unchanged by amiRNAs cloned in the 5'-UTR whereas expression of MyoG was significantly up regulated by ami878 (p=0.0089). However, amiRNAs cloned in 3'-UTR showed significant down regulation of MyoD by 51 (p=0.0007), 27 (p=0.0232), 29 (p=0.0074), and 31% (p=0.0104) and MyoG by 36 (p=0.0034), 12 (p=0.2532), 22 (p=0.0303), and 37% (p=0.0026) by ami204, ami318, ami735 and ami878, respectively. As observed for U6 and CMV driven shRNAs, CMV driven amiRNAs showed significant induction of interferon response in 293T (up to 121.7 fold) and myoblasts (212.5 fold) cells. As ACVR2B has been shown to be essential for embryonic development, we tested the possibility of its knockdown in skeletal muscle using muscle specific MCK promoter. Among the MCK and MSTN promoters with and without two repeats of MCK enhancer, we observed maximum transcriptional activity by MCK promoter in goat myoblasts cells. We thus tested best amiRNAs (ami204 and ami318) by expressing under MCK promoter which showed 22% silencing efficacy by ami318 in 293T cells and 32% silencing efficacy in goat myoblasts cells by transient transfection assay. Further to test the possibility of ACVR2B knockdown after stable integration of amiRNAs into goat myoblasts genome, we generated lentivirus particles carrying amiRNAs expression cassettes and transduced the goat myoblasts. The myoblasts cells stably integrated with amiRNAs showed ~8% silencing by ami318 which was increased to 34% upon induction of differentiation under muscle specific promoter. Western blot analysis revealed 41% and 57% silencing by 5'-ami204 and 5'-ami318, respectively whereas 14% and 35% silencing by 3'-ami204 and 3'-ami318, respectively in stable myoblasts upon induction of differentiation. Unlike transient transfection assay vv'hich showed positive correlation of expression of ACVR2B with myogenic regulators, its stable knockdown resulted in up regulation of MyoD in 5'-UTR derived ami204 and ami318 and overall down regulation of MRFs in 3'-UTR derived ami204 and ami318 integrated goat myoblasts cells. The 5'- UTR derived ami204 and ami318 showed increased rate of cell proliferation as well as myoblasts fusion in stable goat myoblasts compared to scramble control indicating growth promoting effect of ACVR2B knockdown. The skeletal muscle specific partial knockdown of ACVR2B is unlikely to affect the embryonic survival and it will be interesting to further assess its possible growth promoting effect in adult animal by generating transgenic goat through somatic cell nuclear transfer of goat myoblast cells stably integrated with amiRNAs.ThesisItem Open Access MANAGEMENT EFFICIENCY OF ROSE GROWERS(AAU, Anand, 2010) PATEL, DHARMENDRAKUMAR D.; Patel, B. B.The recent advances in rose production technology have demonstrated that scientific management has great potential for increasing the rose production. Therefore, raising management efficiency is of paramount importance for rose producer. This will open up new vistas and make possible for rose growers to achieve substantial gains in income. There are number of factors affecting the rose production. Management is one of the most important factors which help the rose growers to exploit natural resources and accumulate capital. The efficient use of resources depends to a greater extent on how rose growers acquire and adopt innovations in the rose cultivation in effective manner to reach higher levels of economic performance through their management efficiency. The present study on "MANAGEMENT EFFICIENCY OF ROSE GROWERS" was undertaken with following specific objectives: 1. To know the profile of rose growers 2. To study the management efficiency of rose growers 3. To ascertain the relationship of personal, social, economical, communicational and psychological characteristics with management efficiency of rose growers 4. To know the extent of contribution of selected independent variables on dependent variable of rose growers 5. To study the direct and indirect effect of the independent variables on the management efficiency of rose growers 6. To analyze the constraints faced by rose growers regarding various aspects of rose production The present study was undertaken in Dholka taluka of Amdavad district of Gujarat state. There are 71 villages of Dholka taluka. Out of these, six villages were selected based on specific criteria. These selected villages were considered as stratum and with the help of proportional allocation method sample size of each stratum was determined. A random sampling procedure was adopted for the selection of respondents from the selected villages. Thus, total 127 rose growers were selected randomly. The Ex-post-facto research design was followed. A scale was developed to measure the management efficiency of rose growers. The dependent and independent variables were measured with appropriate scales and procedures adopted by other research workers. To measure the knowledge level regarding improved rose cultivation practices and attitude towards improved rose cultivation practices, tests were developed by the investigator. An interview schedule was developed in accordance with the objectives of the study and it was pre-tested and translated into Gujarati. The data of this study were collected through personal interview. The statistical measures such as percentage, mean, standard deviation, co-efficient of correlation, stepwise multiple regressions, standard partial regression coefficient and path analyses were used. MAJOR FINDINGS The important findings of the study are as under: 1. PROFILE OF THE RESPONDENTS 1.1 PERSONAL CHARACTERISTIC Majority of the rose growers had middle age (72.44 per cent), at least primary education level (83.50 per cent) and medium level of experience in rose cultivation (63.80 per cent). 1.2 SOCIAL CHARACTERISTIC Nearly three-fifth (57,50 per cent) of the rose growers had large family size. 1.3 ECONOMIC CHARACTERISTICS More than three-fourth (77.16 per cent) of the respondents possess up to 2.00 hectares of land and a great majority (84.25 per cent) of the rose growers had above 35.00 per cent of their total land under rose cultivation. 1.4 COMMUNICATIONAL CHARACTERISTICS Majority (83.46 per cent) of the rose growers had medium to low level of participation in training programme, more than fourfifth (81,10 per cent) of the rose growers had medium to high level of contacts with extension agencies and more than three-fourth (77.95 per cent) of the rose growers possessed medium to high level of exposure to mass media. 1.5 PSYCHOLOGICAL CHARACTERISTICS Majority (74.80 per cent) of the rose growers had medium to high level of economic motivation, three-fourth (75.59 per cent) of them had medium to high degree of risk orientation, four-fifth (81.10 per cent) of them had medium to high level of achievement motivation, a great majority (89.76 per cent) of them had medium to high level of aspiration, majority (77.95 per cent) of them had medium to high level of innovative proneness, a great majority (85.83 per cent) of them had medium to high level of cosmopoliteness, a great majority (85.83 per cent) of them had medium to high level of competition orientation and a great majority (85.04 per cent) of them had medium to high level of self confidence. 2. MANAGEMENT EFFICIENCY AND ITS COMPONENTS OF ROSE GROWERS 2.1 MANAGEMENT EFFICIENCY OF ROSE GROWERS Slightly more than two-third (68.50 per cent) of the rose growers had medium level of management efficiency, while 16.54 per cent of the rose growers had high level of management efficiency and 14.96 per cent of the rose growers had low level of management efficiency. 2.2 VARIOUS COMPONENTS OF MANAGEMENT EFFICIENCY OF ROSE GROWERS Slightly more than four-fifth (81.10 per cent) of the rose growers had medium to high level of knowledge about improved rose cultivation practices, more than four-fifth (85.83 per cent) of the respondents had medium to high level of attitude towards improved rose cultivation practices, three-fourth (75.59 per cent) of the respondents had medium to high level of ability in planning in rose cultivation, slightly less than four-fifth (79.53 per cent) of the respondents had medium to high level, of ability to make rational decision, majority (83.47 per cent) of the respondents had medium to high level of timely adoption, majority (85.04 per cent) of the respondents had medium to high level of ability to mobilize resources, more than four-fifth (85.04 per cent) of the respondents had medium to high level of ability to co-ordinate activities, slightly less than three-fourth (74.02 per cent) of the respondents had medium level of efficient use of resources, majority (84.25 per cent) of the respondents had medium to high level of ability in rational marketing and slightly less than three-fourth (73.23 per cent) of the respondents had medium to high level of competence in evaluation. 3. RELATIONSHIP OF PERSONAL, SOCIAL, ECONOMICAL, COMMUNICATIONAL AND PSYCHOLOGICAL CHARACTERISTICS WITH MANAGEMENT EFFICIENCY OF THE ROSE GROWERS Out of eighteen independent variables, sixteen viz.; education, experience in rose cultivation, land under rose cultivation, participation in training programme, contact with extension agency, exposure to mass media, economic motivation, risk orientation, achievement motivation, level of aspiration, scientific orientation, innovative proneness, cosmopoliteness, competition orientation and self confidence were positively and significantly related, while age is negatively and significantly related to management efficiency of the rose growers. 4. EXTENT OF CONTRIBUTION OF SELECTED INDEPENDENT VARIABLES ON THE DEPENDENT VARIABLE OF THE ROSE GROWERS Management efficiency was found to be predicted by nine independent variables such as participation in training, scientific orientation, cosmopoliteness, risk orientation, education, land under rose cultivation, level of aspiration, innovative proneness and experience in rose cultivation, which has together contributed 75.80 per cent of the total variation in the management efficiency of the rose growers. 5. DIRECT AND INDIRECT EFFECT OF THE INDEPENDENT VARIABLES ON MANAGEMENT EFFICIENCY OF THE ROSE GROWERS Participation in training programme of the respondents was the key variable in exerting considerably direct and substantial effect on management efficiency. Education and scientific orientation were the major traits in determination of management efficiency through positive indirect effect. Whereas, age of the respondents is only character which negatively and indirectly influenced on management efficiency. 6. CONSTRAINTS FACED BY THE ROSE GROWERS REGARDING VARIOUS ASPECTS OF ROSE PRODUCTION The most important constraints faced by the rose growers regarding various aspects of rose cultivation were: Fluctuation in prices of roses, non-availability of labours for various rose cultivation practices, high cost of inputs, commission rates of agents very high, weed problem, high cost of transportation, lack of technical guidance at proper time, natural calamites/flood and lack of marketing facility near by village.