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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 IMMUNOTOXICOLOGICAL STUDIES OF SUBACUTE ACEPHATE EXPOSURE IN WHITE LEGHORN COCKEREL BIRDS(AAU, Anand, 2006) TRIPATHI, SYAMANTAK MANI; THAKER, A. M.Acephate (Ace), a water-soluble insecticide, belongs to the phosphoramidothioate group of organophosphate (OP) insecticides. Acephate is an organophosphate foliar spray insecticide of moderate persistence with residual systemic activity of about 10-15 days. It is being widely used for the protection of vegetables and fruits due to its activity against lepidopterans and aphids. As this insecticide is in use as crop protectant, it is likely to cause indirect exposure in poultry through contamination of feed, soil and ground water (in very low amount) and hence, the present study was conducted in Day old White Leghorn Cockerels birds; approximate medium lethal dose (ALD50) of Acephate taken into consideration for the study was 852mg/kg. One hundred twenty five birds were divided into five different groups each comprising 25 birds. The birds of group Ci were given no treatment and served as control. Group C2 were administered groundnut oil (Iml/kg) and served as control (vehicle). Group T1 was put onl/40th of ALD50 (21.3 mg/kg), while group T2 received 1/30th of ALD50 (28.4 mg/kg) and group T3 was administered withl/20th of ALD50 (42.6 mg/kg) of Acephate suspended in 1 ml of groundnut oil. Once daily oral dosing was carried out for 28 days. All the birds were monitored for any observable toxic symptoms throughout the experimental period and they were also weighed weekly to monitor body weight gain. The blood samples were collected before sacrificing the birds on day 15 (after 14 days oral dosing) and day 29 (after 28 days oral dosing) of the study and were analyzed for hematological, biochemical and immunological parameters. Organ (liver, spleen, bursa and thymus) weights were recorded and organs (liver, spleen, bursa thymus, lung, and kidney) were collected for histopathological examinations. Severity and extent of the clinical signs varied according to dosage administered to the birds. The clinical symptoms observed were sudden onset of depression, reduced feed intake, dullness, ruffled feathers, cyanosis of comb, green diarrhea and severe limb weakness and some time paralysis. During the study period 6-8 birds died. Nervine symptoms like tremor, head down condition and torticolis were noticed only for few minutes before death. There was a reduction in the body weight of the Acephate treated birds. No alteration had been recorded in haematological parameters (hemoglobin, packed cell volume, total erythrocyte count and total leukocyte count) due to Acephate exposure. Dose dependent significant increase in blood glucose due to administration of Acephate was observed. A significant dose dependent increase in Serum Glutamic Oxaloacetate Transaminase and Serum Glutamic Pyruvate Transaminase and non significant increase in Lactate Dehydrogenase level of the birds treated with the Acephate indicates its systemic effect. A significant decrease in serum proteins during study was observed. A significant decrease in serum albumin and globulin were observed on 28th day of study. During the experimentation, Acephate had dose dependent immunosuppressive effect on Humoral immune response of birds from 28th day of experimentation. These findings indicate significant effect on protein metabolism and humoral immune response at the administered doses of Acephate. Cellular immunity was not affected as tested by DNCB dye test. Present study revealed that Acephate at administered doses seems to be toxic for multiple systems in growing WLH birds. Gross postmortem and histopathological changes in various organs of birds treated with Acephate were observed with typical organophosphate dose dependent toxicity signs. Microscopic changes observed in different organs viz. liver, lung, kidney spleen, thymus and bursa were of typical to organophosphate insecticide poisoning. Though, Acephate has been reported mild to moderately toxic to the birds; it seems that doses of Acephate given in the present study produce mild toxicity to multiple body systems of growing birds including immune system.ThesisItem Open Access ECONOMIC RAISING OF GIR ( Bos indicus ) CALVES BY FEEDING MILK REPLACER(AAU, Anand, 2006) ODEDRA, MULRAJ DEVABHAI; RAVIKALA, K.An experiment of thirteen weeks duration was conducted on eighteen weaned Gir calves randomly divided into three groups of six each. The first group received whole milk feeding, while the second group received milk replacer feeding replacing 25 per cent of milk and the third group received milk replacer feeding replacing 50 per cent of whole milk. Milk replacer was more or less similar to milk in composition and could sustain tiie growth rate of weaned Gir calves as appeared from the acceptability. The results revealed non significant differences in average daily gains under three treatment groups though the periodical changes in body weight were linear in nature. Milk replacer fed groups recorded poor growth compared to whole milk fed group. Changes in biometry were also non significant due to treatment effect. Overall liquid feed consumption, concentrate intake, roughage intake, total DMI (g/day), per cent DMI and DMI per kg metabolic weight did not differ significantly among three treatment groups. Though the consumption pattern of DMI was at par in all the three treatment groups, body weight gain was poor in T-2 and T-3 due to negative growth recorded in the second week of the experiment. Feed efficiency was at par in all the three treatment groups and it was higher in T-1 followed by T-3 and T-2 indicating the superiority of natural milk to that of milk replacer. Overall difference in the cost of feeding/kg body weight gain was also non significant in three treatment groups. Cost of feeding/kg gain was the lowest in T-3 followed by T-1 and T-2.ThesisItem Open Access THE STUDY OF CANINE MAMMARY TUMORS WITH SPECIAL REFERENCE TO MUTATIONS IN p53 TUMOR SUPPRESSOR GENE BY PCR-SSCP(AAU, Anand, 2006) DHAYGUDE, VITTHAL SHRIRANG; Joshi, B. P.The present study entitled "The study of canine mammary tumors with special reference to mutations in p53 tumor suppressor gene by PCR-SSCP" was conducted to know the prevalence of canine neoplasms as well as mammary tumors by analyzing the data available at Department of Pathology, Veterinary College, Anand and to investigate the role of p53 gene in molecular pathogenesis of canine mammary tumors by mutational analysis using molecular technique PCR- SSCP. Epidemiological studies on canine neoplasms with special reference to mammary tumors were carried out by obtaining the autopsy/ biopsy records of last five years (January 2001- December 2004). Information like breed, age, sex and histopathological type was analyzed and classified year wise, age wise and breed wise to acertain the susceptibility of the canine mammary tumors. For study of mutations in the p53 tumor suppressor gene a total of sixteen cases of spontaneously occurring canine mammary tumors were collected by personal visits at clinics, hospitals and Dept. of Surgery at Veterinary College, Anand as well as Bombay Veterinary College, Mumbai with detailed history and gross observations of individual case. The tumor tissues were collected in two sets of vials from each individual case; one in 10% neutral buffered formalin for histopathological examination and other in liquid nitrogen (- 196°C) for DNA extraction and subsequent PCR- SSCP studies for detection of mutations in Exon 5, Exon 6 and Exon 5 to 7 of p53 tumor suppressor gene using gene specific primers. During the period from January 2001 to December 2005, a total of 124 autopsy / biopsy samples of various canine neoplasms were recorded. Among these, tumors of skin and soft tissues were recorded highest 74 (59.67 %) followed by mammary gland tumors 43 (34.67 %), transmissible venereal granulomas 3 (2.4 %) and tumors each of ovary and testes 2 (1.6 %). Among different breeds of canines, these neoplasms were recorded highest in Alsatian 27.41 % followed by Pomeranian 21.77%, Labrador 12.90 %, Mongrels 12.10%, Doberman 8.87 %, Spitz 8.5 %, Great-Dane, Cocker Spaniel, Tibetan Mastiff each of 2.42 % and Daschounds 1.6 %. Majority of tumors (70.15%) were recorded between 5-12 years of age. Overall there were 67.7 % tumors of benign nature while remaining 32.3 % tumors were malignant. A total of 43 mammary gland tumors were recorded during the period January 2001 to December 2005. All the tumors were recorded in female dogs with near equal frequency during last five years. Among different age groups the canine mammary tumors were recorded highest in the age group of 5 to 8 years (41.9%) followed by 9 to 12 years (37.2%), 13 to 16 years (11.6%) and 0 to 4 years (9.3%). Majority of the neoplasms (79.1%) were recorded between 5 to 12 years of age. Alsatian 37.2 % was found to be most susceptible to develop mammary tumors followed by Pomeranian 25.6 %, Doberman 13.9 %, Labrador 9.3 %, Spitz 6.9 % and Mongrels 6.9 %. Histopathologically out of a total 43 mammary tumors 48.8% were of benign nature while remaining 51.2 % were malignant. Benign neoplastic conditions recorded were adenoma simple & papillary (4), fibroadenoma (5) and mixed tumors involving myoepithelial cells, bone, cartilage and fibrous connective tissue (12). Malignant neoplastic conditions were adenocarcinoma solid, tubular and papillary (16) and mixed malignant tumors involving bone, cartilage and fibrous connective tissue (6). Out of a total 16 cases of mammary tumors collected for study of p53 gene mutations; majority of dogs (13/16) developed mammary tumors between the age of 8 to 10 years. Only three were spayed previously while in other 13 cases ovariohysterectomy was done at the time of surgical removal of the tumor. There was variation in size of the mammary tumor ranging between 2 cm X 1.5cm X 1 cm to 21cm X 15cm X 14cm. Shape of the tumors varied from ovoid, elongated, rounded to irregularly nodular. Maximum tumors were hard and firm in consistency. Mammary glands of the right side were found affected more in comparison to left side. The fourth (caudal abdominal) and fifth (inguinal) pairs of mammary gland were found more suscep*:ibie to develop mammary tumor. Histopathologically all the 16 tumors revealed six conditions of benign nature while remaining ten were of malignant nature. The benign neoplastic conditions diagnosed were mixed benign tumor involving myoepithelial cells, cartilage, fibrous connective tissue and glandular epithelial cells (5) and fibroadenoma (1). The malignant neoplastic conditions were diagnosed in 10 cases. They were adenocarcinoma (solid and papillary) in nine cases and squamous cell carcinoma in one case. DNA was extracted from 15 out of total 16 tumor samples by proteinase K and phenol method. PCR amplification of exon 5 and exon 7 of p53 tumor suppressor gene was obtained in all the 15 samples with separate gene specific primers for respective regions. The amplification of exon 6 could not be made by PCR with different annealing temperatures of 50 °C, 55 °C, 60 °C and 66°C. PCR using Hot start Taq Polymerase was also not successful. PCR- SSCP profile revealed aberrantly migrating bands suggestive of mutations in amplified exon 5 (codon 117-175) of p53 tumor suppressor gene in 20% cases of spontaneous canine mammary tumors. Sequencing in one case of papillary adenocarcinoma out of three with mutant bands revealed four mutations. These were represented as two silent mutations; one on codon No. 117 (CTC-> CTT), second on codon 137 (CCA—> CCG) as well as two missense mutations on codon 157 (ACC—>ACA) and codon 158 (GAG—>AAG) which resulted amino acid change from threonine to lysine and glutamine to lysine. All the above changes in nucleotides were heterozygous. Screening of amplified exon 5 (from codon 165) to exon 7 (upto codon 241) of p53 gene segment for detection of mutations with PCR-SSCP revealed similar band pattern for all samples suggesting no mutations when primer E 5-7 was used. The PCR-SSCP studies suggested that mutations in p53 gene might be invariably involved leading to its inactivation in canine mammary neoplastic condition.ThesisItem Open Access USE OF NEOSTIGMINE AS AN ADDITIVE TO LIGNOCAINE AND BUPIVACAINE FOR EPIDURAL ANAESTHESIA IN COW CALVES(AAU, Anand, 2006) KULKARNI, SHEETAL VIVEK; BARVALIA, D. R.A study on epidural anaesthesia was conducted in 24 cow calves allotted to 4 groups of 6 each by injecting the drug(s) into the first intercoccygeal site. In group A, 2% Hgnocaine hydrochloride @ 0.2mg/kg b.wt.; in group B, combination of 2% lignocaine hydrochloride @ 0.2mg/kg b.wt. with neostigmine 250µg; in group C, 0.5% bupivacaine @ 0.1mg/kg b.wt. and in group D, combination of 0.5% bupivacaine @ 0.1mg/kg b.wt. and neostigmine 250µg were injected epidurally through first intercoccygeal site. The total volume for epidural injection was kept constant (2ml) by adding normal saline solution in all the individuals. The efficacy of each drug alone and in combination was evaluated by observing induction time, return of reflex, duration of anaesthesia, heart and respiration rates, rectal temperature at 0, 5, 10, 15, 30 and 60 minutes interval after injection. Onset of anaesthesia was found to be significantly (P < 0.05) early in animals of group D as compared to animals of groups B and C, however, inspite of longer time for onset no significant difference was observed in animals of group A as compared to the animals of group D. Duration of anaesthesia was found to be significantly longer (P < 0.05) in animals of group D as compared to animals of other groups. Moreover, duration of anaesthesia also significantly differ (P < 0.05) in animals of groups B and C as compared to animals of group A, however, no significant difference was observed in animals of groups B and C.Return of pinprick reflex was earliest in the animals when lignocaine HCl was used alone (group A) whereas it was almost similar when lignocaine-neostigmine combination (group B), and bupivacaine alone (group C) were used. However, it was much delayed when bupivacaine-neostigmine combination (group D) was used. Animals developed loss of hindlimb co-ordination i.e. ataxia whenever lignocaine HCl alone (group A) was used, however, all the animals of this group were standing throughout the period of study upto 60 min. post injection, whereas, animals of other groups did not showed any signs of ataxia. Analgesia was restricted only to the base of tail when pin prick reflex made in animals of group D where bupivacaine was used in combination with neostigmine, whereas, analgesia was present on entire tail in animals of other groups. In all the groups, alterations in mean heart and respiration rates and rectal temperature observed at various time intervals were non-significant and remained within the normal physiological limits throughout the period of study. Side effects such as ataxia was seen in some animals wherein lignocaine HCl was used. The present study infers that epidural neostigmine in combination with Hgnocaine HCL and bupivacaine is safe, effective and well tolerated by cow calves and as caudal additive to lognocaine HCl and bupivacaine increased the duration of analgesia vis-a-vis when used alone.ThesisItem Open Access FEASIBILITY OF FEEDING DETOXIFIED CASTOR CAKE (Ricinus communis) TO COMMERCIAL LAYERS(AAU, Anand, 2006) PATEL, D. K.; Parnerkar, SubhashThe experiment was designed to evaluate effects of feeding detoxified castor seed cake at various stages of laying cycle, by simultaneously selecting four age group birds age 20, 34, 51 and 60 weeks. All the age group birds were fed experimental diets for 9 weeks following 3 - days pre-experimental period. A total of 2000 White Leghorn (BV 300) layer birds under each of four age groups i.e. 2000 birds per group were housed in California cages in 668 compartments, each having space for 3 birds and only one bird was housed in last compartment for both the treatment groups for various age groups. One thousand birds were divided into 10 replicates each of 100 birds and in each individual cage three birds of each treatment were placed and only one in last compartment in each age group. Layer birds of four different age groups (1-20 weeks, II- 34 weeks, III-51 weeks and IV-60 weeks) were randomly assigned to two dietary treatment, viz. T1-(control) and T2: 5 per cent detoxified (4 % lime treated and extrusion cooked) castor cake (CCK). The layer mash formulated was isonitrogenous and isocaloric and was offered adlib. The observation on egg production, egg weight, egg quality, feed intake and feed efficiency were recorded for 9 weeks. The metabolic trial was conducted at the end of experiment to find out balance of nitrogen, phosphorus, and calcium and metabolizable energy content of each diet. The economics of feeding was calculated from the record of feed consumption on egg production. The results revealed that the total number of egg / bird in age group (I, II, III and IV) was 43.07±0.47, 55.49±0.55, 50.70±0.52 and 50.60±0.51 in control and 40.7310.45, 53.53±0.53, 50.02±0.51 and 49.16±0.49 in treatment groups, respectively. The means of treatment and control groups did not differ significantly in age group-Ill but differed significantly (P < 0.05) for II and IV and were highly significant (P < 0.01) for age group-I. The hen housed egg production in control group was significantly higher (P < 0.01) in age group-II, III and IV; whereas the hen day egg production was significantly higher (P < 0.01) in age group-II and IV as compared to CCK fed birds. The average egg weight for control birds under 1, II, III and IV age groups was 46.83+2.85, 46.65±1.54, 54.34+0.06 and 53.68±1.45 whereas 53.40±].67, 52.65±l.03, 49.57±2.44 and 54.03±1.03 for treatment birds, respectively. The average egg weight was significantly (P<0.05) higher in treatment group of age group II. In the birds given treatment diet egg weight was, though non-significant, was higher in age group I and IV while lower in age group III as compared to control group. The different egg quality traits viz. Albumin Index, Yolk Index, Haugh Unit and shell thickness were similar in both control and treatment groups. In age group-II the albumin index and shell thickness were significantly (P < 0.05) higher in treatment group. Daily feed intake of birds in age group I, II, III and IV was 103.91 ±0.61, 108.37 ±.83, 113.53 ±0.98 and 115.70 ± 0.90 in control whereas 103.35+ 1.00, 107.59 ± 0.94, 107.94 ± 0.1.00 and 117.66 ± 1.26 in treatment group, respectively were (P < 0.01). The feed consumption / dozen egg produced in group I, II, III and IV was 2.36 ± 0.18; 1.48 ± 0.013; 1.70 ± 0.016 and 1.73 ± 0.016 in control and 2.82 ± 0.28, 1.53 ± 0.014, 1.64 ± 0.016 and 1.82 ± 0.022 in treatment group, respectively. The change was non significant in age group I, significant (P < 0.05) in age group II and highly significant (P < 0.01) in group III and IV. The overall mortality (2.69 %) was very low during the entire experimental period. It was 2.2, 1.2, 7.4 and 2.9 % in control and 1.6, 0.9, 4.3 and 1.0 % in treatment group in age groups I, 11, 111 and IV, respectively. The average cost of feeding (Rs. / dozen eggs) of birds in age group 1, 11, 111 and IV was 15.62 ± 1.16, 9.80 ± 0.09, 11.22 ±0.10 and 11.46 ±0.11 in control and 18.41 ± 1.80, 9.96 ± 0.09,10.69 ± 0.11 and 11.88 ± 0.14 in treatment group, respectively which was significantly (P < 0.05) higher in age group III and IV whereas non significant in age group I and II. The average balance of nitrogen ranged between (0.24±0.021 and 0.41±0.03), calcium (0.48±0.015 and 0.88±0.23) and phosphorus (0.12±0.04 and 0.29±0.13) were more or less similar in all the four age groups in control and treatment groups. The metabolizable energy consumed also did not differ significantly in age groups I, III and IV, but was higher (P < 0.05) in age group-II of treatment group as a result of high dry matter intake. Thus, the inclusion of detoxified (4 % lime treated and extrusion cooked) castor cake at 5 per cent level in layer mash had no any adverse effect on over all performance of caged layer birds and can be used as alternate feed resource without any economic benefit.ThesisItem Open Access STUDIES ON CRYOSURGICAL OOPHORECTOMY IN BITCHES(AAU, Anand, 2007) MANKAD, MANAN YOGESHBHAI; TANK, P. H.Cryosurgical oophorectomy (Group - I; n=6) and conventional surgical oophorectomy (Group - II; n=6) following standard midline laparotomy were compared in a clinical trial on twelve healthy bithces. The ovaries were cryofrozen to -20° C by liquid nitrogen spray under cryogaurd protection employing double freeze-thaw cycles. The cryosurgical oophorectomy was bit quicker (31.53 ± 0.9 minutes) as compared to conventional surgical oophorectomy (34.24 ± 0.64 minutes). There was significant fall in the rectal temperature following both the methods of oophorectomy. Additionally, there was mild but nonsignificant fluctuations within physiological range in the physiological parameters viz., pulse rate, respiratory rate and haemoglobin oxygen saturation as a combine effect of general anaesthesia and surgical and cryosiirgical procedures. The haemoglobin concentration and packed cell volume did not differ significantly in the animals during perioperative observations but it showed significant difference between the animals of both the groups. The variation in the erythrocyte sedimentation rate remained nonsignificant during perioperative observations in the animals of both the groups. The total erythrocyte count fluctuated nonsignificantiy within the physiological range at different time intervals, but this variation was significant statistically between the animals of both the groups. The nonsignificant variation of total leucocyte count observed between the animals of both the groups indicated that none of the procedures has specific influence on this vital cell count. The neutrophils rose initially up to 48 hours, which then got substituted by the lymphocytes. Hence, an almost reverse trend was observed for the lymphocyte during the present study. A transient post-operative elevation in serum Cortisol level following the procedures in both the groups of animals with tendency to resume normalcy at later phases was suggestive of temporary pain and / or stress imposed by surgical as well as cryosurgical oophorectomy. Cryosurgery remained uneventful during post-operative observations with absence of systemic reflections, which indicated local damage limited to the frozen ovaries only. Laparoscopic as well as histomorphological assessment of cryodamaged ovaries a month later confirmed their complete nonvitality. The cryosurgical oophorectomy was found simple, quick, bloodless, stressfiree, safe and equally effective as compared to conventional surgical oophorectomy. Additionally, it was cost-effective as well as its clinico-surgical execution was feasible.ThesisItem Open Access GENETIC ANALYSIS OF PRODUCTION AND REPRODUCTION TRAITS OF TRIPLE CROSS (HF 25 % X J 25 % X KANKREJ 50 %) CATTLE ON AN ORGANIZED FARM(AAU, Anand, 2006) PATEL, JITENDRAKUMAR SHANTILAL; Rank, D. N.Present study was undertaken for genetic analysis of production and reproduction traits of HJK triple cross (HF 25% x J 25% x Kankrej 50%) maintained at Livestock Research Station, AAU, Anand, Gujarat from 1990 to 2005. The study included analysis of 572 lactation records of 223 HJK triple cross cows of three generations belonging to 36 sires. Data were scrutinized for abnormality and finally 266 lactation records of 126 cow progenies of 21 sires having complete set of all necessary records were used for genetic analysis. Various traits like Calf weight (CW), Weight at first calving (WFC), Age at first calving (AFC), Service period (SP), Calving interval (CI), Breeding efficiency (BE), Lactation length (LL), Dry period (DP), Lactation milk yield (LY), Standard lactation milk yield (SLMY), Lifetime total lactation days (LTTLD), Lifetime total milk yield (LTTMY), Average milk production per day of lactation length (MY/LL), Average milk production per day of calving interval (MY/CI), Average milk production per day of lifetime milking days (MY/PL), Average milk production per day of lifetime calving intervals (MY/HL), Peak yield (PY), Days to attain peak milk yield (DAPMY), Persistency of lactation. Milk production efficiency per kg of WFC (MPEK), Milk production efficiency per kg of WFC and lactation days (MPEKD), genetic and non-genetic factors affecting the traits, lactation curve dynamics and path analysis have been studied. Data were analyzed using Mixed Model Least Squares and Maximum Likelihood computer programme (Harvey, 1990) to overcome the non-orthgonality of the data. Three sets of data for F1, F2 and F3 generations were categorized. Various effects like sire, season of calving, period of calving, parity, etc. on important dairy traits were estimated by using model-1, genetic parameters like heritability, genotypic and phenotypic correlation, etc. were estimated by using model-2 and repeatability was estimated by using model-8. The Gamma, Inverse Polynomial, Gaines and Rook functions were fitted to derive lactation curves using SPSS programme. The Path analysis was carried out using phenotypic correlations of traits affecting SLMY. The least squares means of SLMY, PY, DAPMY, MY/LL, MY/CI, LL, DP, SP, CI, AFC, Birth weight, WFC, BE, MPEK, MPEKD, TC, TCI, TLTLD, TLTMY, MY/PL and MY/HL in Fl generation of HJK triple cross were 2253.76 ± 41.31 kg, 13.91 ± 0.24 kg, 27.96 ± 1.47 days, 7.49 ± 0.15 kg, 5.53 ± 0.13 kg, 299.63 ± 3.68 days, 115.56 ± 7.3 days, 137.74 ± 7.11 days, 415.20 ± 7.12 days, 1032.95 ± 27.40 days, 22.93 ± 0.19 kg, 335.55 ± 14.99 kg, 86.64 %, 6.43 ± 0.92 kg, 0.021 ± 0.0019 kg, 3.61 ± 0.64 unit, 1498.15 ±-268.97 days, 1079.39 ± 195.50 days, 7436.61 ± 634.13 kg, 4,96 ± 0.25 and 2.93 ± 0.22 respectively. The estimates for the same in F2 and F3 generations were 2166.68 ± 47.16 and 1846.90 ± 47.34 kg, 12.21 ± 0.24 and 09.56 ± 0.63 kg, 30.98 ± 3.42 and 16.50 ± 1.39 days, 7.19 ± 0.15 and 6.30 ± 0.23 kg, 5.64 ± 0.15 and 5.11 ± 0.14 kg, 308.94 ± 4.07 and 287.00 ± 4.46 days, 94.97 ± 7.99 and 66.50 ±2.08 days, 125.99 ±^.16 and 85.00 ±4.59 days, 403.92 ± 8.20 and 353.50 ± 3.69 days, 1227.65 ± 31.78 and 1285.60 ± 93.42 days, 22.42 ± 0.13 and 21.99 ± 0.29 kg, 314.09 ± 8.33 and 294.59 ± 17.50 kg, 90.09 and 99.17 %, 6.87 ± 0.52 and 6.41 ± l.Ol kg, 0.019 ± 0.0011 and 0.019 ± 0.0022 kg, 3.42 ± 0.4 and 3.32 ±0.74 unit, 1378.26 ± 168.16 and 1179.96 ± 208.98 days, 1053.36 ± 118.06 and 952.84 ± 125.53 days, 6685.15 ± 452.91 and 6207.74 ± 896.12 kg, 4.85 ± 0.16 and 5.26 ± 0.46 & 2.56 ± 0.18 and 2.51 ± 0.34 respectively. Their pooled estimates were 2207.61 ± 31.25 kg, 13.08 ± 0.18 kg, 28.98 ± 1.68 days, 7.34 ± 0.11 kg, 5.57 ± 0.09 kg, 303.30 ± 2.69 days, 105.71 ± 1.09 days, 131.56 ± 2.04 days, 409.02 ± 6.02 days, 1148.11 ± 51.02 days, 22.34 ± 0.21 kg, 322.96 ± 4.70 kg, 88.40 %,6.66 ± 0.20 kg, 0.02 ± 0.0003 kg, 3.5 ± 0.13, 1421.67 ± 81.06 days, 1059.94 ± 40.17 days, 6990.43 ± 342.51 kg, 4.91 ± 0.09 and 2.71 ± 0.14 respectively. There was decline in SLMY (18.05 %), PY (31.28 %), MY/LL (13.89 %), MY/CI (7.60 %), LL (4.02 %), WFC (12.21 %), MPEK (0.32 %), MPEKD (9.53 %), TLTLD (11.77 %), TLTMY(16.53 %) on interbreeding. AFC increased by 24.5 % and DAPMY by 41.00 %. However, there was improvement in BE (15.26 %), MY/PL (8.45 %) and MY/HL (7.96 %). Improvement was also observed in the form of decreased DP (42.45 %), SP (38.0 %), CI (14.96 %) and TCI (21.24 %). The effect of generation was significant only for SLMY, CI, MY/LL, PI-1, PI-4 and PI-7. The season of calving affected significantly SLMY, LY, SP, CI, MY/LL, MY/CI, BE, PI-I, PI-4, PI-6 and PI-7. The period of calving had significant effect on SLMY, LY, SP, CI, MY/CI, DAPMY, PI-2, PI-3 and PI-8. The parity had significant effect on SP, MY/LL, PY and PI-5. The sires contributed significantly in SLMY, LY, CI, MY/LL and PI-5.ThesisItem Open Access ASSESSMENT OF NOVEL N-GENE TARGET FOR DETECTION OF PESTE DES PETITS RUMINANTS (PPR) VIRUS BY RT-PCR AND CHARACTERIZATION BY RFLP PROFILE(AAU, Anand, 2006) CHANDRA, VARTIKA; Jhala, M. K.Peste des Petits Ruminants (PPR) is an economically important viral disease of sheep and goats, characterized by high fever, ocular and nasal discharge, pneumonia, necrosis and ulceration of the mucous membrane and inflammation of gastrointestinal tract leading to severe diarrhea. The disease is caused by a Morbillivirus of the family Paramyxoviridae. The disease is highly contagious causing varying degree of morbidity and mortality in susceptible animals. In India, severity of the disease is more pronounced in goats than in sheep and with a combined susceptible population of about 200 million, it is one of the major threats to the small ruminant population of the country. The present study was aimed to study the incidence of Peste des petits ruminants virus (PPRV) in goats of selected areas of Gujarat by sandwich ELISA and to derive estimates of overall, locationwise, agewise and sexwise incidence and samplewise positivity rates. In addition, the study also involved standardization and application of novel Nucleocapsid (N) gene based RT-PCR, for diagnosis of PPR. Further, an attempt was also made to access possible genetic variation among the field and vaccine PPRVs by RFLP profile. A total of 46 clinical samples from 31 goats suspected of PPRV from selected areas of Gujarat were tested by sandwich-ELISA, of which 23 animals were found positive yielding an overall incidence rate of 74.19 percent. Out of 18 animals suspected of PPR at Clinical Service Complex, Veterinary College. Anand, 15 (83.33%) were found positive; whereas four (80%) out of five samples from Koth, two (66.67%)) out of three samples from Vasna and two (40%) out of five samples from Harsoli yielded positive results. Out of the 31 animals, 13, 11, 6 and one animals belonged to age groups of 0-12, 12-24, 24-48 and >48 months, respectively, showing the respective incidence of PPRV as 84.61, 63.63, 66.67 and 100 per cent. Incidence rate of PPR by sandwich-ELISA was 80%) and 71.43%) in males and females, respectively. The PPRV antigen could be detected slightly more in blood samples (73.33%) than in nasal swabs (66.67%o), in case where both the sample types were taken from the same animals. All the 46 clinical samples, a reference vaccine virus (Sungri isolate) and a blood sample from apparently healthy goat were processed for RNA extraction using TRI Reagent . RNA samples showing acceptable purity and concentration were reverse transcribed with random hexamers to generate cDNA templates, which were subjected to PCR amplification using in silico designed N-gene specific primer pair N1-N2. Reference vaccine virus as well as fifteen (32.61%) of the 46 clinical samples, including fourteen blood samples and one nasal swab, produced the desired amplion of approximately 463 bp with primer pair N1-N2. The amplicons from all the positive samples and the vaccine virus migrated similarly in the gel. Thirty one (67.39%)) samples including 12 blood samples, 17 nasal swabs and two tissue samples as well as the blood sample taken from apparently healthy goat failed to produce the targeted amplification with the primer pair N1-N2. Out of 46 clinical samples tested for PPRV, 31 and 15 samples were positive by sandwich-ELlSA and RT-PCR, respectively. "None of the samples negative in sandwich- ELISA yielded positive amplification in RT-PCR. Relative to sandwich-ELlSA, sensitivity and specificity of the RT-PCR was 48.39 and 100.00 per cent, respectively. Overall agreement between the two tests was 65.22 per cent. RT-PCR products (463 bp) amplified with primer pair N1-N2, were further digested with two restriction enzymes viz. Pst-I and Alu-I for RFLP analysis. RT-PCR products of the representative samples from each of the three locations and the reference vaccine virus produced similar RFLP patterns with each of the two REs, indicating absence of genetic variation at the corresponding restriction sites of these two REs. Digestion of RT-PCR products with Alu-l yielded a fragment of 395 bp, whereas the other fragment was not visible. Similarly, digestion with Pst-I yielded a single fragment of405bp.ThesisItem Open Access EPIDEMIOLOGICAL STUDIES OF CANINE NEOPLASMS WITH SPECIAL REFERENCE TO MUTATIONAL ANALYSIS OF p53 GENE IN CANINE MAMMARY TUMORS BY PCR-SSCP(AAU, Anand, 2006) JIVAN, PAWAR SANJAY; Ghodasara, D. J.Epidemiological studies of canine neoplasms with special reference to mutational analysis ofp53 gene in canine mammary tumors by PCR-SSCP were carried out to know age, sex and breed wise incidence of neoplastic conditions in canine and to study the mutations in exon 4 and exon 8 of p53 gene segment in canine mammary tumors. The epidemiological study was conducted by analyzing available data of last ten years (1996- 2005) of 175 specimens of canine neoplasms at Department of Pathology, Veterinary College, Anand and PCR-SSCP based mutational analysis p53 gene segment was carried out on 16 canine mammary tumor specimens collected at Department of Surgery, Veterinary College, Anand and various clinics during surgical removal. The mutations were further confirmed by direct sequencing of PCR products by using Gene specific primers. During last ten years (1996-2005), Department of Pathology, Veterinary College, Anand has received 175 specimens of canine neoplasms for histopathology/ biopsy examinations. Data analysis of 175 specimens revealed occurrence of neoplastic conditions more in female (65.14%) than male (34.85%) dogs. Highest risk of development of various tumors was observed at age group between 7-9 years of age followed by 10-12 years, 4-6 years and 0-3 years. Frequency of occurrence of neoplasms was observed more in Alsatian breed (25.71%) followed by Pomerian (22.85%), Doberman (12%), Labrador (10.28%) and Mongrel (6.85%). Benign neoplastic conditions were observed in 51.42%o cases while malignant neoplasms were observed in 48.57 % cases. Rate of malignancy was noted higher in Alsatian, Pomerian and Doberman breed whereas, Labrador and Mongrel breed showed majority of benign cases. Age group between 4-12 years shows majority of malignant cases and mammary gland tumors in females was deciding factor. Maximum malignancy was observed in the age group between 7 to 9 years of age followed by 10 to 12 years. Adenocarcinoma was the most frequently observed neoplasm followed by fibroma, transmissible veneral granuloma, basal cell carcinoma and squamous cell carcinoma. Skin, mammary gland and genital organs were the most common sites for the neoplasms. The tumors of the muscles, bones, urinary tract and gastrointestinal tract showed relatively low incidence. Clinical diagnosis of 16 mammary gland tumors used for p53 gene mutational analysis showed more incidences in Alsatian breed followed by Pomerinian and Doberman. Majority of the mammary tumors were occurred in the age group between 7- 10 years (14/16). All of the tumors were observed in adult bitches. Risk of mammary tumor was greater in non spayed bitches (13/16) as compared to spayed bitches (3/16) indicating that hormonal imbalance may be major risk factor. Gross morphological examination of mammary tumors revealed majority of the tumors as spherical, ovoid or round and were nodular with hard consistency, some of which were contained inflammatory exudate. Fourth and fifth pair of mammary glands were the common mammary tumors occurring lands. The size of tumor was ranging from peanut to coconut size measuring from 1 to 12cm in diameter. Histopathological examinations showed six conditions of benign nature while remaining 10 were of malignant nature. Among the benign tumors, mixed tumor involving myoepithelial cells, cartilage, fibrous connective tissue and glandular epithelial cells were in five cases with single case of fibroadenoma. In malignant neoplasms adenocarcinoma (solid and papillary) and squamous cell carcinoma observed in nine and one case respectively. The exon 4 and exon 8 of p53 tumor suppressor gene was investigated for mutations in 16 spontaneous canine mammary neoplasms using PCR-SSCP with direct sequence analysis of PCR products. Genomic DNA was successfully extracted from 15 out of 16 tissues. PCR fragments having expected size of 233 bp were obtained from all 15 DNA samples. But PCR failed to amplify the target sequence with primer exon 8 inspite of using appropriate PCR conditions. Fifteen samples were screened for mutation detection on exon 4 of the p53 tumor suppressor gene by PCR-SSCP. Analysis revealed that 5 (30.3%) of 15 canine mammary tumors displayed aberrantly migrating bands indicative'of a p53 gene mutation. Variation in migration pattern in SSCP bands was obtained in sample No. 1 (adenocarcinoma), 5 (myoepithelioma), 7 (benign mixed), 13 (myoepithelioma) and 14 (papillary adenocarcinoma). Mutations detected by PCR-SSCP were further confirmed by sequence analysis in 3/5 samples. Sequence analysis of 3 tumor samples revealed 2 mutations in exon 4. One missense mutation on codon 106 (GCC—>GGC) was found in all three samples. Whereas Sample No. 13 presented one silent mutation at codon 58 (GAT—»GAA). The silent mutation in sample No. 13 was due to heterozygosity at third nucleotide (T) on codon 58. Mutations were present on highly conserved domain of p53 tumor suppressor gene suggesting importance of the region in apoptosis. These results demonstrated the involvement of p53 gene mutations in the development of the canine mammary tumors.