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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 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 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.ThesisItem Open Access TOLL LIKE RECEPTORS (TLR) EXPRESSION IN MILK SOMATIC CELLS DURING MASTITIS IN CATTLE USING REAL-TIME PCR(AAU, Anand, 2011) GHADIGAONKAR, DINESH DILIP; Rank, D. N.The dairy sector in India has shown remarkable progress in the recent years and consequently the country has now become the largest milk producer and valueadded milk products in the world. Though the country is the largest producer of milk, milk production per animal is much less compared to agriculturally developed countries. One of the reasons for this less milk production per animal is loss of milk production capacity because of inflammation of the mammary gland or mastitis in dairy animals. Mastitis is defined as an inflammatory reaction of the parenchyma of the mammary glands to bacterial, chemical, thermal or mechanical injury which is characterized by a range of physical, chemical and usually bacteriological changes in the milk and pathological changes in the glandular tissue which is the most common and the most expensive disease of dairy cattle throughout the world. Mastitis is classified as subclinical and clinical mastitis. The cows that have subclinical mastitis are reservoirs of organisms that lead to infection to other cows. Most clinical cases start as subclinical; thus, controlling subclinical mastitis is the best way to reduce the clinical cases. Somatic cells consist of leukocytes and epithelial cells exfoliated from the mammary epithelium. Mastitis is associated with an influx of inflammatory cells; hence somatic cell count of milk increases. Enumeration of somatic cell counts and bacterial culture of milk has been suggested as a standard method for detecting subclinical udder infections in dairy cows. Genes have major impact on health status of animals. Genetic variability of mastitis resistance is well established in dairy cattle. Resistance to mastitis is a complex function involving various biological pathways, molecules and cells. Study of the expression of genes involved in mastitis resistance is major tool for early diagnosis of disease and genetic improvement to superior stock. Toll Like Receptors (TLR) are cell-surface receptors that recognizes a broad class of pathogen-associated molecular patterns, activates immune system, and induces the over expression of inflammatory factors, which participate in irmate immune responses to confer disease resistance. The bovine TLR genes have been studied in recent years. Hence the study was planned with the objective to investigate expression of TLR-2, TLR-4, TLR-9 in somatic cells in healthy and mastitic udder by Real Time PCR in cattle. The study was undertaken to assess different TLRs (TLR-2, TLR-4 and TLR- 9) in three cattle breeds namely Gir, Kankrej and Triple crossbred (Kankrej x Jersey x Holstein Friesian) with sub-clinical mastitis. A total of 65 lactating cows comprising 16 Gir, 29 Kankrej and 20 Triple crossbred animals were screened for presence of mastitis using Electronic Somatic Cell Counter and bacteriological culture examination. Total RNA was extracted from milk somatic cells from 15 positive and 6 healthy quarters from each breed using TRIZOL method. The RNA was treated with DNase enzyme to remove any traces of genomic DNA. cDNA was synthesized from RNA using Qiagen's Omniscript reverse transcriptase kit and random hexamer primers. The amplification of cDNA template of TLR 2, TLR 4 and TLR 9 genes was carried out using gene specific primers. Expression of TLR 2, TLR 4 and TLR 9 genes mRNA was quantified by Real Time PCR and analysed using Applied Biosystems 7500 SDS software. Relative expression study of these genes was carried out using GAPDH as internal control. Results indicated that there was upregulation of TLR 2, TLR4 and TLR9 gene expression in animals affected with subclinical mastitis compared to healthy animals. Targeted amplification of 421 bp TLR 2, 108 bp TLR 4 and 108 bp TLR 9 was confirmed by agarose gel electrophoresis. Prevalence of subclinical IMI was higher in Triple crosbred cows (65%) compared to Gir (50%) and Kankrej cows (27.59 %). The mean SCC of infected quarters was significantly higher than that of noninfected quarters (P < 0.05) in all three breeds. The average relative expression of all three genes i.e. TLR2, TLR4 and TLR9 was higher (ranged from 7 to 35 folds) in mammary gland with subclinical intramammary infection than those measured in the uninfected glands. The concomitant increase in somatic cell count and upregulation of TLR2, TLR4 and TLR9 gene expression was observed during subclinical mastitis in all three breeds. Comparison of SCC upregulation between breeds indicated that, there was no significant difference between breeds in the SCC in the diseased quarter during subclnical mastitis in Gir, Kankrej and triple crossbred cows. In Gir cows, TLR2 gene expressions level in diseased quarters was found to be upregulated with an average 10.54 (10.54 ± 7.12) fold compared to pooled healthy quarters. In Kankrej cows, TLR 2 gene expressions level in diseased quarters was found to be upregulated with an average 12.22 (12.22 ± 11.61) folds compared to pooled healthy quarters. In Triple crossbred cows, TLR 2 gene expression level in diseased quarters was found to be upregulated with an average 7.13 (7.13 ± 10.57) folds compared to pooled healthy quarters. In Gir cows, TLR 4 gene expressions level in all diseased quarters was found to be upregulated with an average 18.43 (18.43 ± 24.230) fold upregulation compared to pooled healthy quarters. In Kankrej animals, TLR 4 gene expressions level in diseased quarters was found to be upregulated with an average 31.59 (31.59 ± 18.74) folds compared to pooled healthy quarters. In Triple crossbred cows, TLR 4 gene expression level in diseased quarters was found to be upregulated with an average 23.817 (23.817 ± 27.6963) fold compared to pooled healthy quarters. In Gir cows, TLR 9 gene expression level in diseased quarters was found to be upregulated with an average 6.193 (6.193 ± 8.19) fold compared to healthy quarters. In Kankrej cows, TLR 9 gene expression level in diseased quarters was found to be upregulated with an average 5.44 (5.44 ± 8.14) folds compared to Pooled healthy quarters. In Triple crossbred cows, in all diseased quarters, TLR 9 gene expression level was found to be upregulated with an average 19.29 (19.29 ± 16.31) fold compared to pooled healthy quarters. During subclinical mastitis SCC was found to be positively correlated with the transcriptional activities of TLR2, TLR4 and TLR9 gene in Gir and Triple crossbred cow. In Kankrej cows TLR 2 and TLR 9 gene expressions were positively correlated with s e c but TLR 4 gene expression was not correlated with SCC. The level of infection as reflected by number of somatic cells had significant effect on level of upregulation in gene expression. However, there was no significant effect of a breed on level of upregulation of TLR 2, TLR 4 and TLR 9 gene expression.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.ThesisItem Open Access CLINICAL STUDIES ON ILIZAROV TECHNIQUE FOR IMMOBILIZATION OF TIBIAL FRACTURES IN DOGS(AAU, Anand, 2010) Bhatia, Ami Sandip; TANK, P. H.A clinical study on Ilizarov technique for immobilization of tibial fractures in dogs was carried out with multiple objectives of knowing the epidemiological status of various orthopaedic affections, standardizing the application of 'Ilizarov technique' for external immobilization of tibial fractures on cadaver models and lastly, applying and comparing the Ilizarov technique with intramedullary pinning for immobilization tibial fractures in dogs. Last three years' retrospective information was retrieved by scrutinizing the data bank of the College Hospital. During that period, a total of 8790 canine cases were registered out of which 486 (5.53%) cases were having orthopaedic maladies. Amongst those, the cases of fractures were 378 that accounted for nearly 78% cases of the reported orthopaedic maladies. Majority of the fractures involved radio-ulna (29.36%) followed by femur (28.30%), tibia-fibula (23.80%) and humerus (14.55%). Cadaveric tibiae were used for standardization of the Ilizarov technique. The diaphyses of tibiae were sawed in a manner to mimic different fractures and these fractures were immobilized using Ilizarov technique of circular external fixation to get hands on training. To evaluate the feasibility of the Ilizarov technique in the treatment of tibial diaphyseal fracture in dogs, a study was carried out in 12 clinical cases of tibial fractures. The cases were randomly divided into two groups based upon the fracture stabilization technique used, viz., Ilizarov technique (Group I, n=6) and Intramedullary pinning (Group II, n=6). No exudation was seen in group I whereas mild exudation was observed in few cases of group II. Partial weight bearing was seen as early as 2nd postoperative day in group I as against 7th postoperative day in group II. Complete weight bearing was observed from the 6th postoperative day in majority cases of group I as compared to 24th postoperative day in group II. The earlier restoration of the functional usage of the limb could be attributed to the closed reduction, no external infection and axial stability of the Ilizarov apparatus. In majority cases of group I joint mobility was earlier, seen from the 2nd post operative day whereas limited joint mobility was seen in the first week which gradually improved in all the cases of group II. Radiological examination was done at weekly intervals. Most of the dogs belonging to both the groups showed proper reduction and good alignment of the bone fragment immediately after the operation. Callus fomiation was earlier i.e. within 45 days in majority cases of group I as against 54 days in all cases of group II. One case of group I did not show any bridging between the fractured fragments. Minimum or no periosteal reaction was seen around fracture site in group I whereas considerable periosteal reaction was seen around the fracture site in group II. Mild periosteal reaction was seen around the K wire insertion sites in group I by 4th postoperative week. Postoperative complications observed in group I were mild wire tract infection which subsided with proper antibiotic dressing. In group I, majority of cases healed completely except one case in which the animal took off the apparatus within 15 days of operation. Also, one case did not show bridging between the fractured ends. This could have been due to preoperative infection at the fractured site. Few cases showed mild wire tract infection. In group II, occasional limping was observed inspite of complete weight bearing. This might have been due to irritation to soft tissues around the entry site of Steinmann pin while flexing and extending the stifle joint. Infection was seen in one case of group II at the incision site and mild rotation of the distal fragments were seen in few cases. Removal of the Ilizarov apparatus or pin was carried out at 5-7 weeks in group I and 7-8 weeks in group II. This was done when clinical healing was observed in the fomi of functional limb use and endosteal and/or periosteal bridging was evident on radiographs. The Ilizarov apparatus, using external circular transosseous fixation provided better anatomic reduction, axial stability of the fractured fragments, greater joint mobility and functional weight bearing. This technique also prevented muscle atrophy and rotational instability unlike intramedullary pinning. Hence Ilizarov technique is better suited for treating tibial fractures than intramedullary pinning.ThesisItem Open Access ISOLATION, IDENTIFICATION AND MOLECULAR CHARACTERIZATION OF BACTERIAL PATHOGENS FROM RESPIRATORY TRACT OF APPARENTLY HEALTHY AS WELL AS SICK GOATS(AAU, Anand, 2011) AHER, TUSHAR KISAN; ROY, ASHISHA number of factors are responsible for economic losses to the goat industry; among them the pneumonia due to various bacterial species imposes serious constraints on goat production all over the world because of high mortalities. The major health problem of small ruminants is pneumonia/pleuropneumonia, which may be caused by Mycoplasma and Pasteurella species alone or in conjunction with other microbes. Pneumonia in small ruminants constitutes a serious setback to the growth in this group of animals with resultant economic losses in many parts of the world. Thus, the present study was undertaken with a view to know preponderance of this bacterial spp. in relation to respiratory tract infections in apparently healthy and sick goats. The objectives were isolation, identification, nucleic acid based detection of virulence associated and toxigenic potentials and in vitro antibiotic sensitivity patterns of the isolates from respiratory tract infections of apparently healthy as well as sick goats. In the present investigation, total 102 nasal swab samples and 96 tissue samples were collected from apparently healthy as well as sick goats. Bacterial isolation was done following standard technique by inoculating tissue sample and nasal swab sample primarily on blood agar and plates incubated for 24-48 hrs at 37°C. After incubation, the nature of growth and cultural characters of colonies were studied. Preliminary morphological identification was based on Gram's staining. Specific identification and biochemical characterization of the isolates was done as per the standard techniques. In this study, ten different types of bacteria were isolated. It includes Mycoplasma spp. (0.7%), P. inultocida (0.7%), Staphylococcus spp. (29.9%), Micrococcus spp. (4.2%), Streptococcus spp. (9.7%), Bacillus spp. (19.4%), E. coli (18.8%), Proteus spp. (4.9%), Klebsiella spp. (5.6%) and P. aeruginosa (6.3%). The most prevalent species of bacteria found was Staphylococcus spp. Gram positive organisms were more prevalent in apparently healthy goats (46.5%) than sick goats (11.8%). Gram negative organisms were more prevalent in sick goats (24.3%) than apparently healthy goats (16.7%). From 102 nasal swab samples- 68 isolates, 32 lung samples- 31 isolates, 32 trachea samples- 26 isolates, 32 tonsil samples- 19 isolates were obtained. Out of which. Gram positive bacteria were 91 (63.2%), whereas Gram negative bacteria was 52 (36.1%) and a single isolate was identified as Mycoplasma spp. (0.7%). From 102 nasal swabs, total 68 isolates were obtained and there were total nine different types of bacteria isolated, viz.. Mycoplasma spp. (1.5%), P. multocida (1.5%), Staphylococcus spp. (38.2%). Micrococcus spp. (8.82%), Streptococcus spp. (7.4%), Bacillus spp. (33.8%), E. coli (5.9%), Klebsiella spp. (1.5%) and P. aeruginosa (1.5%). The most pre\alent bacterial species found in nasal swab were Staphylococcus spp. From. 32 lung samples, total 31 isolates were obtained and there were total seven different types of bacteria isolated, viz., Staphylococcus spp. (19.4 %). Streptococcus spp. (12.9%), Bacillus spp. (6.5%), E. coli (35.5%), Proteus spp. (6.5%), Klebsiella spp., (9.7%) and P. aeruginosa (9.7%). The most prevalent bacterial species found in lung was E. coli. From 32 tracheal samples, total 26 isolates were obtained and there are total six different types of bacterial species were isolated. It involves Staphylococcus spp. (30.8 %), Streptococcus spp. (7.7%)), Bacillus spp. (11.5%), E. coli (34.6%), Proteus spp. {1.1%) and P. aeruginosa (1.1%). The most prevalent bacterial species found in trachea was E. coli. From 32 tonsillar samples, total 19 isolates were obtained and there are total six different types of bacterial species were isolated. It involves Staphylococcus spp. (15.8 %), Streptococcus spp. (15.8%), E. coli (15.8%o), Proteus spp. (15.8%), Klebsiella spp., (21.1%) and P. aeruginosa (15.8%)). The most prevalent bacterial species found in tonsil was Klebsiella spp. Molecular characterization of the isolates by PCR based method was applied for specific detection as well as detection of virulence associated and toxigenic genes.