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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 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 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.ThesisItem Open Access EPIDEMIOLOGICAL AND EXPERIMENTAL STUDIES ON ETIOLOGY OF VISCERAL GOUT IN BROILER CHICKS(AAU, Anand, 2005) Khodabhai, Patel Ashvinkumar; Ghodasara, D. J.Gout is a disorder of uric acid metabolism that allows uric acid to accumulate in the blood and tissues. Uric acid is the end product of protein and purine metabolism and is produced normally by the body during tissue remodeling and breakdown. The present epidemiological and experimental study was carried out to know the incidence of visceral gout in field condition and role of high protein, high calcium, water deprivation and diclofenac drug for causation of visceral gout as well as the role of allopurinol drug for treatment of visceral gout in broiler chicks. The available data on poultry autopsies from Department of Pathology, Veterinary College, Anand for a period of six years were analysed for epidemiological study. Based on autopsy records, incidence of visceral gout in broiler chicks up to three weeks of age was found to be 9.98 per cent with a yearly frequency of 6.52 to 19.50 per cent. Out of total cases of visceral gout recorded, 65.23 per cent (autopsy incidence) cases were recorded during colder months. The highest autopsy incidence of 94.99 per cent was recorded in Cobb breed of broiler. Of the susceptible age of first three weeks, the highest autopsy incidence of 57.03 per cent was recorded during the second week of age followed by 21.48 per cent each in third and first week of age. The experimental studies were carried out by conducting the two experiments. Experiment-I was conducted to know the role of marginal high protein (27%), high calcium (1.6%), water deprivation (for first 36 hours and than drinking water was given for one hour every day) and diclofenac drug (2.5 mg/kg body weight/day) for causation of visceral gout as well as the role of allopurinol drug (15mg/kg body weight/day) for treatment of visceral gout in broiler chicks. For this experiment 210 day old broiler chicks were divided in to seven different groups with different treatments and samples were collected on 3rd, 6th 9th 12th and 15th days of age from each group. Experiment II was carried out to know the effect of diclofenac drug at various durations for causation of visceral gout in broiler chicks. Diclofenac was given orally at the dose of 2.5 mg/kg body weight/day for one, three, seven and fifteen days in group II, III, IV and V respectively. Blood samples and kidney tissue were collected from all the. birds on fifteenth days for plasma uric acid and histopathology. — The plasma uric acid concentrations were increased on 6th day onward with high protein diet (27 %). Histopathological investigation revealed mild congestion and haemorrhages of kidney parenchyma, degeneration of tubular epithelium and lymphoid aggregation at 12th day onward in high protein group. Increased level of plasma uric acid with high protein diet came to normal level when allopurinol (15 mg/kg body weight/day) treatment was given in high protein + allopurinol group. In water deprivation group the plasma uric acid concentrations were increased significantly at ninth day onward. The kidney body weight ratio gradually increased from first to fifth collection in water deprivation group. In high calcium fed group the concentration of plasma uric acid was increased three times at fifteenth day of age. The concentration of plasma creatinine was also significantly increased at 15th day of experiment. In high calcium group^ the microscopic lesions of the kidney were characterised by mild to moderate congestion and haemorrhages of kidney parenchyma, tubular degeneration, inter- tubular fibroblastic proliferation and focal infiltration of mononuclear cells. Diclofenac caused hyperuricemia and mortality due to visceral gout in broiler chicks by oral administration. The plasma uric acid concentration increased gradually from collection I to collection V in diclofenac treated group in experiment-I. The plasma uric acid levels in experiment II were also increased in ascending order according to the duration of diclofenac treatment. In experiment II maximum mortality due to visceral gout was observed in group V (12%) followed by group IV (8%) and III (4%). The lesions were characterised by varying degree of white chalky deposition on the serosal surfaces of visceral organs. Histopathologically radiating pattern of urate crystals were seen in the kidney parenchyma.ThesisItem Open Access PATHOLOGiCAL AND SEROLOGICAL STUDIES ON RESPIRATORY INFECTIONS IN COMMERCIAL BROILER CHICKS(AAU, Anand, 2004) Girdharbhai, Rank Chirag; Joshi, B. P.Epidemiological, serological and pathological studies on respiratory infections were carried out in ten commercial broiler farms in and around Anand district of Gujarat State. To know the prevalence of respiratory infections, autopsy records of the last three years i.e. Jan. 2001 to Dec. 2003 were analyzed. The available data of autopsy and farm mortality records were classified year wise, month wiss, age wise and breed wise to ascertain the susceptibility and incidence of disease. A total of 1,565 flocks were affected with respiratory infections out cf 3,940 flocks received for autopsy. The relative incidence of respiratory infections based on autopsy records was found to be 39.72 per cent with year wise frequency ranging from 36.50 to 42.49 per cent. Among these, the condition complicated chronic respiratory disease was recorded at highest rate of 90.86 per cent. There was meagre incidence of Ranikhet disease, aspergillosis and pneumonia. Month Vi^ise autopsy records showed higher incidence (45.17%) during the summer season. Among the affected flocks the highest incidence of 50.99 per cent was recorded in avian-34 breed of broilers followed by hubbard and hubchix constituting 46.61 and 41.57 per cent respectively. Age wise incidence showed tiiat tiie iiigliest number of cases at the rate of 55.62 per cent were recorded during 3rd week followed by fourth week (47.75%) and fifth week (43.98%) of age. Studies of the farm incidence at ten broiler farms totalling the population of 68,378 broiler birds showed an average mortality rate of 4.38 per cent with a range of 2.26 to 7.98 per cent. Farm wise mortality due to respiratory infections varied between 0.16 to 5.80 per cent with an average mortality rate of 1.86 per cent. Among tnese the condition colisepticaemia was recorded at the highest rate of 95.29 per cent. Week wise farm mortality showed the highest incidence of 0.64 per cent during the third week of age followed by fourth (0.57%), fifth (0.35%) and sixth (0.24%) week of age. Breed wise farm mortality due to respiratory infections showed the higher incidence in Hubbard (1.35%) in comparison to cobb (0.51%o) breed of broiler out of a total mortality of 1.86 per cent due to respiratory infections. The overall seroprevalence of infectious bronchitis (IB) was found to be 18.52 per cent (100/540 sera sample). The antibodies against IB were detected in 6 out of 10 farms (60.00%)). The individual farm wise seroprevalence ranged between 0.0 to 100 per cent. All the sera sample were negative for Mycoplasma gallisepticum antibodies. The Micro HI titres of Ranikhet disease in most of the farms were within the protective antibody levels. Escherichia coli organisms were isolated from all the 60 samples of colisepticaemia representing all the ten broiler farms. The important gross lesions observed in colisepticaemia were congestion of mucosa of nostrils, trachea and bronchi covered by thick mucous exudate. In majority of cases airsacculitis, presence of caseous plugs in the lumen of trachea and bronchi, fibrinous pericarditis and fibrinous perihepatitis were noticed. Lesions observed in aspergillosis were presence of grayish white nodules of varying sizes and numbeis on the lung parenchyma. The histopathological lesions in colisepticaemia were of the nature of variable degree of congestion, vacuolar degeneration and desquamation of epithelial cells and mild to moderate infiltration of heterophils and mononuclear cells in the tracheal and bronchial mucosa. The lesions in the lung parenchyma were comprised of mild to moderate oedema, mild to severe congestion, focal to diffuse haemorrhages, focal to diffuse infiltration of hererophils and mononuclear cells and presence of caseous exudate in the lumen of parabronchi. Air-sacs revealed mild oedema and heterophil infiltration with fibroblast proliferation and accumulation of caseous exudate. Liver and heart revealed thickening of the liver capsule and pericardium with deposition of fibrinous exudate along with mononuclear cells infiltration. Hepatic and cardiac parenchyma showed mild to severe congestion and haemorrhages, focal to diffuse areas of necrosis and focal to diffuse infiltration of mononuclear cells. The lesions in the lung were comprised of mild to moderate congestion, hyperplasia of reticulo-endothelial cells with lymphoid cell depletion and focal presence of bacterial colonies. On the basis of serological observations, isolation of Escherichia colt organisms as well as gross and microscopic lesions, it may be opined that the predominant respiratory infection is colisepticaemia which is of systemic involvement and may exist either alone or aggrevated in the presence of primary infection such as infectious bronchitis virus causing heavy mortality.ThesisItem Open Access ETIOPATHOLOGICAL STUDIES ON VISCERAL GOUT IN BROILER CHICKS(AAU, Anand, 2001) Shrivastava, Nidhi; Joshi, B. P.ThesisItem Open Access STUDIES ON INFECTIOUS BURSAL DISEASE VIRUS INDUCED IMMUNOSUPPRESSION AND ITS EFFECT ON LYMPHOID ORGANS IN LAYER CHICKS(AAU, Anand, 2001) A D, Deshmukh; Joshi, B. P.The present study was undertaken to evaluate IBDV induced immunosuppressive effect on different lymphoid organs i.e. bursa of Fabricius, spleen, thymus, caecal tonsils and gland of Harder in layer chicks. A primary survey in relation to autopsy incidence of the disease was also undertaken from the available data on poultry autopsies from January 1996 to December 2000 to ascertain the severity and importance of the disease condition. The relative incidence of infectious bursal disease (IBD) was found to be 9.74 per cent with yearwise frequency varying from 7.70 to 12.39 per cent. Majority of these cases (70.7%) were recorded during summer months (March to July). The mortality was recorded in the age group of 2nd to 16th week with peak incidence between fourth to ninth week of age. Among different breeds of layer birds the frequency of the disease was found to be highest in Babcock layers followed by Bovans, Rhode Island Red (RIR) and Lohman. Experimental induction of IBDV induced immunosuppression was carried out by procuring 140, day-old White Leghorn Babcock layer chicks and after dividing them into two equal groups as-control and infected on 14th day of age. The infected group of birds were inoculated witli bursal homogenate prepared from natural outbreaks of infectious bursal disease. Ten birds from each group were sacrificed at each weekly interval for the period of five weeks (7 to 35 day PI) and detailed study was made for the evaluation for set parameters. Experimental IBDV Infection resulted in significant rise in IBD ELISA antibody titres and significant fall in NDHI titres at each PI interval upto 35 day PI. The serological response to NDV vaccination was found to be reduced at each PI interval. Boosting with LaSota vaccination on 28* day of birds age (14 day PI) could not result in improvement of titres. There was significant reduction in bursa : body weight ratio (BF:BW) at 14, 21, 28 and 35 day PI and no alteration in spleen body weight (S:BW), thymus body weight (T:BW) and gland of Harder : body weight (GH:BW) at any post-infection interval. Total leukocyte count fall significantly while differential leukocyte count showed significant rise in heterophils and significant fall in lymphocytes throughout the period of experiment. Clinical signs of mild dullness, depression, ruffled feathers and white diarrhoea along with twelve per cent mortality was noticed during acute phase of the first week post-infection period. Gross lesions were confined to BF and marked by reduction in its size to its pronounced atrophy with the advancement of age. Histopathological changes in the BF were progressive in nature. The lesions became severe and more marked with progression of the experiment. At 7 day PI, mild to moderate depletion of lymphoid cells, cystic spaces in the bursal follicles giving moth eaten appearance and interfollicular heterophilic and mononuclear cells infiltration were the prominent lesions. The lesions were more marked at 14 day PI with diffuse necrosis of bursal follicles, hyperplasia and hypertrophy of corticomedullary epithelium and extensive fibrous tissue proliferation. In subsequent post-inoculation interval of 21 and 28 day PI, the lesions became still severe with extreme small size of follicles, cystic dilatation of mucosal epithelium and extensive fibroplasia. On 35 day PI bursal lesions were marked by extensive fibrous tissue proliferation, extreme small sized follicles, and glandular pattern formed by proliferation of mucosal lining epithelium. Few of the bursal follicles were seen regenerated during this stage in some of the birds. Mild lesions in the form of mild depletion of lymphocytes and RE cell hyperplasia were observed in spieenic parenchyma and caecal tonsils at 7 and 14 day PI. Gland of Harder showed depletion of lymphoid cell and plasma cell population compared to the control birds at 21, 28 and 35 day PI. The fields IBDV isolate resulted in its transmissibility by inducing immunosuppressionThesisItem Open Access SUBACUTE ORAL TOXICITY STUDY OF FEBUXOSTAT IN SPRAGUE DAWLEY RATS(AAU, Anand, 2009) PATEL, VIJAYKUMAR B.; Prajapati, K. S.Febuxostat is a non-purine analogue and is a selective inhibitor of oxidized and reduced forms of Xanthine Oxidase that significantly reduces serum uric acid levels. The present research work was conducted to evaluate the repeated dose toxicity of febuxostat in Sprague Dawley rats. The animals were divided in 5 different groups each included 6 male and 6 female animals. They were administered febuxostat at 0 (vehicle only), 2, 10, 50 and 10 (Recovery) mg/kg, orally for 28 days respectively. The animals were observed for clinical signs and growth parameters. Hematological, biochemical, urine analysis, necropsy, organ weight and histopathological studies were conducted. Oral administration of febuxostat did not produce any toxic symptoms and there was no any change in feed consumption, body weight and body weight gain as compared to control group. No significant hematological alterations were noticed up to 10 mg/kg dose in both sexes. Leucocytosis with neutrophilia was noted in both sexes at 50 mg/kg dose group. There was increase in urea, creatinine and phosphorus along with decrease in calcium and sodium in the serum of male and female from dose group of 50 mg/kg. The dose dependent reduction in serum uric acid level was observed in animals of both sexes from group II, III, IV and V. Urine was yellowish and turbid in the animals of dose group 10 mg/kg and 50 mg/kg. Microscopic examination showed presence of erythrocytes and amorphous crystals in the male and female animals of dose group 50 mg/kg. The significant increase in relative weight of kidney was found in both the sexes of high dose group only. Gross pathological changes observed in the kidney of both the sexes in high dose group were paleness, flabbiness, enlargement and pinpoint whitish necrotic foci on the surface. Urinary bladder in this group showed distention with presence of yellowish urine with fine granular material. Histopathological changes were seen only in kidneys and urinary bladder of animals from group III and IV. Major histopathological changes observed in the kidneys were hyaline deposits in kidney tubules, basophilia of tubules, thickening of glomerular basement membrane, cystic dilatation of tubules, infiltration of inflammatory cells in intertubular space, proliferation of interstitial cormective tissue, neovascularization, necrosis of tubular epithelium, presence of necrosed inflammatory cells in tubular lumen, deposition of crystals in collecting tubules and papillary duct and papillary epithelial hyperplasia. The drug febuxostat was found effective in reducing serum uric acid levels in normal SD rats and was found nephrotoxic at dose levels of 10 and 50 mg/kg. The NOAEL was 2 mg/kg in SD rats.ThesisItem Open Access STUDIES ON COMPATIBILITY OF TIAMULIN, CHLORTETRACYCLINE AND TYLOSIN WITH SALINOMYCIN IN BROILER CHICKENS(AAU, Anand, 2006) Paneliya, Sanjaykumar Mansukhlal; Joshi, B. P.The present study was conducted on day old commercial broiler chicks to know the synergistic and adverse effects related to combination of drugs tiamulin, chlortetracycline and tylosin as antimycoplasmal agents when used along with salinomycin as feed treatment as anticoccidial agent for 42 days. The experiment was conducted in two replicates comprised of five groups with 50 birds in each experimental group. All the groups were fed salinomycin 12 % at the rate of 60 ppm from day one to forty two days of experiment. The groups A1 and A2 (A) were given tiamutin 45% (Tiamulin) at the rate of 5 gm per 200 liters of water for 1-21 days and at the rate of 10 gm per 200 liters of water for 21-42 days in drinking water and chlortetracycline 15 % at the rate of 670 mg per kg of feed for 1-42 days. The groups B1 and B2 (B) were given tylan soluble 100% at the rate of 5 gm per 1000 birds for 1-3 days and at the rate of 45 gm per 1000 birds at 20th day in drinking water. The groups C1 and C2 (C) which served as control group and were given only salinomycin 12% at the rate of 60 ppm in feed. The groups D1 and D2 (D) as well as E1 and E2 (E) were given tiamutin 45% (Tiamulin) at the rate of 1 gm per 2.3 liters of water for 1-3 days and at 21 -22 days respectively in drinking water. The weekly water and feed consumption, weekly body weight, average daily gain and feed conversion ratio were recorded. The daily observations for any clinical signs and mortality were recorded. The blood samples were collected on 3rd, 22nd and 42nd day from each experimental group for biochemical estimations of AST, LDH and CK. Detailed post mortem examination was performed on birds belonged to group D on 3rd day and group E on 22nd day of both the replicates. On day 42, birds from groups A, B and C were euthenised humanely and observed for any gross pathological lesions and microscopic changes if any. There was marked reduction in weekly water and feed consumption in groups D and E i.e. tiamulin treated groups of both the replicates. The marked reduction in water and feed consumption was also observed in group A which was given additional chlortetracycline from fourth and fifth week onwards respectively in comparison to only salinomycin treated group C as well as tylan treated group B. There was no difference in water and feed consumption between tylan treated group B and only salinomycin treated group C during the period of experimentation. Among the different experimental groups FCR was found better in salinomycin treated group C as well as tylan treated group B in comparison to tiamulin treated groups A, D and E during both the replicates. The weekly mean live body weights showed significant reduction in experimental groups A, D and E of both the replicates at the end of 6th week as compared to groups B and C. The clinical signs were only observed in the groups D and E of both the replicates. They were observed within 24 to 48 hours after tiamulin water therapy and were of the nature of sudden depression, drastic reduction in water and feed consumption, difficult movement, lethargic, sternal recumbency with leg weakness. huddle together and paralysis of leg and wing. Amongst this paralysis and huddling were only observed in group D1 and D2. The clinical signs were gradually disappeared after discontinuation of tiamulin containing water. There was no treatment related mortality observed in all the groups and mortality of 1-2 birds in each group was of routine nature i.e. yolk sac infection and colibacillosis. The enzymes AST, LDH and CK showed similar pattern of alteration. The increased levels of all the three enzymes were observed in experimental groups A, B and D in comparison to control group C on 3rd day of experiment. On day 22nd and 42nd elevated levels were seen in all tiamulin as well as tylan treated groups A, B, D and E as compared to control group C. The elevated levels of these enzymes might be due to hepatic cell, skeletal muscle or other soft tissue damage. Gross pathological lesions were observed only in birds which belonged to experimental groups D and E which sacrificed on day 3rd and 22nd respectively. They were of the nature of mild to marked paleness of pectoral muscles. The internal organs like liver, heart, spleen and kidney did not reveal any gross pathological alterations in any groups during the study. The pectoral muscle lesions were correlated with time of drug given i.e. during 1-3 days and 21-22 days as well as higher dose levels of tiamulin. The histopathological lesions observed in liver parenchyma were of the nature of mild to moderate congestion, mild to severe fatty changes, focal infiltration of mononuclear cells and mild perivascular connective tissue proliferation. These lesions were observed with more severity in tiamulin treated groups and indicated its toxic effect while undergoing metabolism in the liver. The severity of hepatic lesions was less in tylan treated group and suggested that tylan water therapy is more compatible than tiamulin water treatment in the presence of salinomycin in feed as anticoccidial agent. The lesions in heart observed during the study were only of the nature of mild congestion and found not of much significance. The splenic lesions in different experimental groups were of the nature of mild to moderate congestion and mild RE cell hyperplasia. The lesions were observed with varied severity in the entire experimental group including control and assumed that they might be in response to birds immunological response in combating the antimicrobial agents. The renal parenchyma showed mild to moderate congestion, focal areas of intertubular haemorrhages, mild to moderate tubular vacuolar degeneration, cystic tubular dilatation and mononuclear cells infiltration. The overall nature of renal lesions gave impression that renal tubular epithelium could be damaged with tiamulin and tylan water medication. The pectoral muscles showed lesions only in the experimental groups D and E of both the replicates. These were of the nature of swelling of muscle fibers, hyalinization of muscle fibers and destruction of sarcoplasm. The lesions suggested that tiamulin treatment is not compatible when salinomycin is used in feed as anticoccidial agent. The present study provided the guidelines to the broiler farmers that tiamulin therapy is not recommended when salinomycin is used in the feed as anticoccidial agent. However further research is needed to decide dose level of tiamulin and chlortetracycline combination along with salinomycin which can prevent adverse effect on performance as well as control the mycoplasma infection which is very much prevalent in the broiler birds.ThesisItem Open Access STUDY ON PATHOGENICITY OF DICLOFENAC SODIUM ON SWISS ALBINO MICE (Mus musculus)(AAU, Anand, 2009) BHAJANTRI, BASAVARAJ S; Prajapati, K. S.The present research work was conducted on 24 male and 24 female Swiss albino mice to study the repeated dose toxicity of Diclofenac sodium. Animals were randomly divided into 4 different groups with six male and six female in each. Group I severed as control. Group II, III and IV were administered with 2.37, 4.75 and 9.5 mg/kg of Diclofenac sodium per kg body weight respectively once daily for 28 days. The animals were monitored for any observable toxic symptoms and mortality throughout the experimental period. All the animals were weighed weekly to monitor body weight gain. The food consumption was also measured once a week. After completion of 28 days treatment, blood samples were collected for haematology and serum biochemical analysis. Necropsy was performed in all sacrificed animals and gross lesions were recorded. Tissue samples 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 dullness, depression, lethargy, and rough hair coat were noticed in mice of group III and FV. The dose dependent reduction in body weight and feed consumption was observed in animals of mid and high dose group. There was significant reduction in the red blood corpuscles, packed cell volume, haemoglobin and mean corpuscular haemoglobin concentration, where as significant increase in mean corpuscular volume and absolute neutrophils in animals of group III and IV. Differential leucocyte count revealed significant increase in neutrophil and decrease in lymphocyte count in animals of Diclofenac sodium treated groups in and IV. In treatment group III and IV, the AST, ALT and Total bilirubin values were significantly increased. Where as significant decrease in total protein, albumin, globulin, and cholesterol was noticed. There was significant increase in urea, creatinine and uric acid in group in and IV. The animals of group IV revealed significant decrease in absolute organ weights of liver, kidneys and there was increase in absolute weight of spleen. Gross pathological changes were characterized by congestion and hemorrhages in intestine and stomach. Enlargement of spleen was the common findings in the animals of group in and IV. Varying degrees of degeneration was noticed in histopathological sections of liver and kidney. There were erosions, ulceration and necrosis of mucosal epithelium of stomach and intestine. Spleen showed hyperplasia of RE cells while lung showed mild congestion and edema in animals of group III and group FV. Gross and microscopic lesions were indicative of gastrointestinal and renal toxicity. The results of the present study indicated toxic potential of Diclofenac sodium, if used for longer time in animals.