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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 SUBACUTE TOXICOPATHOLOGICAL STUDIES OF TAMOXIFEN IN WISTAR RATS(AAU, Anand, 2011) PANCHAL, VIJAY P.; Ghodasara, D. J.The present research work was conducted on 24 male and 24 female Wistar rats to study the toxicoathological effects of repeated dose (28 days) of tamoxifen. Wistar rats were randomly divided into 4 different groups with six males and six females in each group. Animals of group II to IV were given 1, 10 and 20 mg/kg b.wt tamoxifen by oral gavage for 28 days where as group I was administrated only 0.5% CMC as (vehicle) control. After completion of 28 days treatment, blood samples were collected for haematology and serum biochemical analysis from retro-orbital plexus with the help of capillary tube. The animals were sacrificed by high dose of anesthesia with Di - ethyl ether on 29th day for necropsy and collection of tissue. Necropsy examination was performed in all sacrificed animals and gross lesions were recorded. Tissue samples (lung, liver, kidney, intestine, spleen, testes, epididymis, heart, brain and uterus) were collected in 10% formalin solution for histopathological examination. The extent and severity of observed symptoms varied according to the dosage administered to animals. Symptoms like weakness, loss of appetite, aggressiveness and mild alopecia were noticed in rats of high dose group. The dose dependent reduction in body weight and feed consumption were observed in animals of group II, III and IV. The significant decrease in RBC count, packed cell volume, haemoglobin and MCV was recorded in group IV whereas significant increase in total leucocyte count was noticed in group III and highly significant increase in group IV animals. The differential leucocyte count revealed significant increase in neutrophil count in group III and highly significant increase in group rV animals whereas significant decrease in lymphocyte count in animals of tamoxifen treated group IV. No significant change in monocyte, eosinophil and basophil counts were observed in tamoxifen treated groups. AST and ALT values increased significantly in group III and highly significantly in group IV. The significant increase in AKP, creatinine and BUN values were recorded in treatment group IV. The significant decrease in total protein and albumin were observed in treatment group HI and highly significant decrease in group IV. All the rats exposed to tamoxifen at three different dose levels revealed dose dependant pathological changes in group III and IV in different organs. The lesions were characterized by degeneration, necrosis, inflammatory and vascular changes. The main target organs affected were liver, testes and uterus. The overall lesions gave impression that tamoxifen was hepatotoxic as well as toxic to reproductive system. The intensity and distribution of such lesions were more severe in rats of group TV followed by group III.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 Effect of foliar application of plant growth regulators and micronutrients on growth, yield and quality of acid lime cv. Kagzi (Citrus aurantifolia Swingle)(AAU, Anand, 2012) MADHUKAR, JAGTAP VAIBHAV; Patel, H. C.The present investigation on "Effect of foliar application of plant growth regulators and micronutrients on growth, yield and quality of acid lime cv. Kagzi (Citrus aurantifoUa Swingle)" was carried out during spring season of the year 2011 at Horticultural Research Farm, Department of Horticulture, B. A. College of Agriculture, Anand Agricultural University, Anand. The results revealed that treatment ZnS04 0.5 % + FeS04 0.5 % produced significantly the maximum tree height (3.45, 3.73 and 3.98 m at 4, 6 and 8 month after first spray). East West canopy spread (3.44, 3.76 and 4.04 m at 4, 6 and 8 month after first spray), South North canopy spread (3.40, 3.70 and 3.97 m at 4, 6 and 8 month after first spray). Treatment GA3 50 mg/1 took significantly minimum number of days for fruit set after first spray (23.67 days) while significantly minimum number of days for fruit maturity after first spray (135.67 days) were recorded with NAA 200 mg/1. Simultaneously, GAs 50 mg/1 also showed significant increased in tree height, East West canopy spread, South North canopy spread as well as also minimized number of days for fruit set. Yield attributing characters like fruit volume (47.90 cc), fruit diameter (4.54 cm), fruit weight (47.40 g) and fruit yield per tree (46.38 kg) significantly increased under treatment GA3 50 mg/1. In case of number of fruits per tree treatment NAA 200 mg/1 significantly increased the number of fruits per tree (1020.33) and it was statistically at par with treatments 2,4-D 20 mg/1, ZnS04 0.5 % + FeS04 0.5 %, NAA 100 mg/1, GA3 50 mg/1, ZnS04 0.5 % and FeSO4 0.5%. Quality attribiiting characters like total soluble solid (9.58 Brix.) and ascorbic acid content (30.41 mg/lOOg pulp) were significantly increased while number of seeds per fruit (6.13) and acidity (7.05 %) were significantly decreased under treatment GA3 50 mg/1. However, in case of total soluble solid and number of seeds per fruit, it was statistically at par with treatments GAs 50 mg/1, NAA 200 mg/1 and 2, 4-D 20 mg/1, ascorbic acid content was at par with treatment ZnS04 0.5 % + FeS04 0.5 %, NAA 200 mg/1, ZnS04 0.5 %, FeS04 0.5 %, 2, 4-D 20 mg/1 and GA3 25 mg/1 and acidity % was statistically at par with treatment NAA 200 mg/1, ZnS04 0.5 % + FeS04 0.5 %, 2, 4-D 20 mg/1, ZnS04 0.5 % and FeS04 0.5%. From economical point of view, the maximum net realization of ? 89,664 ha with BCR of 1:2.41 was obtained in treatment NAA 200 mg/1 due to increased in fruit retention of plant followed by treatment GA3 50 mg/1 with net realization of ? 87,379 ha and BCR of 1:2.30 due to increased in fruit weight and size. It can be concluded that for securing higher fruit yield (46.00 kg), net return (Rs 89,664) and cost benefit ratio (1:2.41) with quality fruits the treatment NAA 200 mg/1 perform best and next best treatment was found GA3 50 mg/1.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 STUDIES ON BALANCED ANAESTHESIA USING BUTORPHANOL - ACEPROMAZINE - GLYCOPYRROLATE (BAG) AS PREANAESTHETIC TO KETAMINE - DIAZEPAM, KETAMINE -MIDAZOLAM, PROPOFOL AND ISOFLURANE MAINTENANCE IN CANINES(AAU, Anand, 2010) KAVECHIYA, VIPULKUMAR PRAHLADBHAI; BARVALIA, D. R.A clinical study, on 36 dogs of either sex allotted to six groups of six animals in each, was undertaken to evaluate sedative effect of BAG - Butorphanol, Acepromazine and Glycopyrrolate @ 0.2, 0.04 and 0.01 mg/kg b.wt., respectively, as a combination administered intramuscularly. Induction was carried out using Diazepam @ 0.28 mg/kg b.wt. and Ketamine @ 5 mg/kg b. wt. mixture in KDK (I) and KDI (II) groups, Midazolam @ 0.28 mg/kg b.wt. and Ketamine @ 5 mg/kg b.wt. mixture in KMK (III) and KMl (IV) groups and Propofol @ 5 mg/kg b.wt. in PP (V) and PI (VI) groups intravenously (IV) about 15 min. after premedication with BAG. The maintenance of anaesthesia was carried out with ketamine (0.25 mg/kg/min.) in ketamine-diazepam and ketamine-midazolam induction for KDK and KMK groups. Animals of similar induction groups were also maintained with isoflurane in KDI and KMI groups. In propofol induction, propofol (0.4 mg/kg/min.) and isoflurane were used as maintenance agent in PP and PI groups. Anaesthesia was induced in overnight fasted and preanaesthetically evaluated dogs 15 min. after premedication over a period of 90 sec. in ketamine combination groups and over 90-120 sec. time in propofol groups. Onset of sedative effect of preanaesthesia, induction quality and time of intubation were recorded in all animals. Optimum doses for desired effect were also recorded for preanaesthesia, induction and maintenance in all animals. Different clinical parameters related to anaesthesia, recovery times, vital signs and complication, if any, were recorded at 15 min. after preanaesthesia (AP), 0 min. (at induction), 5, 10, 15, 30, 45, 60 and 90 min. inclusive of commencement of induction in all groups. Different haematological and biochemical parameters were studied at 0 min. (normal), 15 and 30 min. post induction in all cases. Cost of preanaesthesia, induction and maintenance were calculated for all cases. Postoperative pain managmant was achieved using meloxicam (0.2 mg/kg b.wt.) for 5 days after operation. All the animals showed profound signs of sedation without any complications. Combination mixture (BAG) also masked the adverse effects of opioid in preanaesthesia. Lower dose of acepromazine seem to be safe for older dogs also. Induction of anaesthesia was smooth and satisfactory without any complications, however it was comparatively faster in propofol and ketaminemidazolam groups than ketamine-diazepam. Time for induction and endotracheal intubation was also less in propofol and ketamine-midazolam induction than ketamine-diazepam. In all six groups, mean pulse rate per minute showed a nonsignificant decrease at 15 minutes after preanaesthesia. At the time of induction and after induction, mean pulse rate showed significant increase in KDK, KDI and KMK groups and nonsignificant increase in KMI, PP and PI groups. Mean Sp02 values showed nonsignificant decrease after preanaesthesia and followed by increase in all groups. However, increase was observed in animals of isoflurane maintenance groups. Respiratory rate showed nonsignificant changes through out the period of anaesthesia. Same way the rectal temperature decreased gradually but nonsignificantly in all groups. Mean blood pressure (systolic, diastolic, mean) showed nonsignificant decrease after preanaesthesia in all groups there after increased nonsignificantly in ketamine maintenance groups but remained in normal range in all groups. There was nonsignificant increase in AST and ALT values in all groups but minor increase in PP, PI, KDI and KMI groups. BUN and creatinine values decreased nonsignificantly in all groups but minor decrese in PP, PI, KDI and KMI groups. Blood glucose concentration showed nonsignificant increase in all groups whereas the total protein concentration were decreased nonsignificantly in all groups. TEC decreased nonsignificantly in PP and PI groups whereas there was nonsignificant increase in all other groups. TLC, haemoglobin and PCV decreased in all groups however was nonsignificant. In DLC, neutrophils increased and lymphocytes decreased nonsignificantly in all groups. The changes in monocytes, eosinophils and basophils were not significant. Recovery characteristics like recovery start time, time for response to pin prick, head raising time, sternal recumbency time, stand unassisant time and resume to feeding or watering time were significantly decreased in PP and PI groups. The cost of propofol and ketamine-midazolam inductions were significantly higher as compared to ketamine-diazepam. Maintenance cost of anaesthesia with isoflurane and propofol were cheaper as compared to ketamine. Meloxicam provided optimum postoperative pain relief All the protocols were competitively effective having no adverse effects on animals. Propofol in combination with isoflurane for maintenance in BAG premedicated group was cheaper and the safest protocol.ThesisItem Open Access STUDY ON BLOOD BIOCHEMISTRY IN RELATION TO MILK PROTEIN WITH SPECIAL REFERENCE TO KAPPA CASEIN IN INDEGENOUS GOAT(AAU, Anand, 2010) PATEL, SANJAYKUMAR B.; Pande, A. M.Caprine casein genes exiiibit an unusual and extensive polymorphism that affects milk quality and composition. The Kappa casein gene includes 5 exons, 4 of them carrj'ing more than 90 per cent of the information to encode for the mature protein. Caseins comprise about 80 per cent of the total protein content of milk and present polymorphism with changes in the amino acid sequence. Within this abundance of proteins. Kappa casein is noteworthy, since it has been associated with differences in milk yield, composition and processing. The objective of this study was to observe the existence of polymorphism in the Kappa casein gene and to compare the levels of plasma and milk biochemical parameters in lactation stage of Zalawadi goats. For this purpose, blood samples from 50 lactating goats, collected with vacutainer by needle puncture of the jugular vein. Plasma separated from blood and cellular parts (WBCs) was used for genomic DNA extraction. Milk samples were collected in glass test tube (50 ml capacity) v^ith boric acid preservative and milk samples were preserved at -20 °C till laboratory analyses. The milk biochemical profiles were estimated by using the ECOMILK analyzer and milk urea nitrogen and total casein were estimated by Auto method and kjeidahl method, respectively. The blood biochemical profiles were estimated by using the BS 120 CHEMISTRY analyzer. Plasma sodium and potassium were measured by Flame photometer. DNA was extracted from cellular part of blood. Goat Kappa Casein (CSN3) gene (exon 4) was amplified from genomic DNA samples by PCR. The PCRs were carried for the amplification of Kappa casein {CSN3) gene with specific primers (F: - 5' TCC CAA TGT TOT ACT TTC TTA ACA TC 3') and (R: - 5' GCG TTG TCC TCT TTG ATG TCT CCT TAG 3') gene loci. Amplified products were visualized as a single compact band of expected size of 645bp under UV light by gel documentation system. PCR products and restriction enzymes were incubated overnight in water bath at 37 °C for digestion with Haelll and Alw441 RE and at 65 °C for digestion with BseNI RE. Digested products were visualized as per restriction patterns with compact band size under UV light by gel documentation system. There were monomorphic restriction patterns with HaeIII-RFLP digestion, whereas polymorphic restriction patterns shown with BseeNI-RFLP and Alw44I-RFLP. In this study, genotypes AA, AG, GG ware observed with BseNI and CC, CT, TT with Alw44I in these animals except HaeIII RFLP. The frequency of allele A and allele G was 0.87 and 0.13, respectively and the frequency of allele C and allele T was 0.88 and 0.12, respectively. There were non-significant and positive correlations between blood glucose with milk fat, plasma total proteins with milk protein and milk casein and plasma total cholesterol with milk fat. There was non-significant and negative correlation between plasma triglycerides with milk fat. There was highly significant (P<0.01) and positive correlation between blood urea nitrogen and milk urea nitrogen.