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2018 - 20195
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Start date: 2010 × End date: 2019 × Subject: Veterinary Pathology ×
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    ThesisItemOpen Access
    “STUDIES ON MAJOR RESPIRATORY PATHOGENS OF BROILER CHICKENS IN AND AROUND JUNAGADH” 2824
    (JAU, JUNAGADH, 2019-06) CHHATROLA MAYUR HARJIVANBHAI; D. T. Fefar
    The present research work was carried out to determine the etiological agents responsible for respiratory distress with special reference to presence of LPAI (H9N2), IB virus, ND virus and E. coli infection in broiler birds. The study comprised of epidemiological information in relation to farm wise mortality and age wise mortality, gross and histopathological examination of involved respiratory organs especially trachea, bronchi, lung and air sacs, detection of LPAI (H9N2) virus, IB virus and ND virus by RT-PCR, detection of avian pathogenic E. coli by PCR and also isolation as well as antibiogram against E.coli from the birds affected with respiratory distress. Studies on the farm incidence and age wise mortality among twenty five broiler flocks having total population of 75,800 birds of second to fifth week of age showed an average mortality rate of 3.25 percent ranging between 1.33 to 8.00 percent. The week wise mortality was found highest during third week of age followed by second, fourth and fifth weeks of age. Gross pathological lesions were predominantly found in organs of respiratory system i.e. in trachea, bronchi, lungs and air sacs. The tracheal lesions observed were severe congestion, excessive mucous exudate, presence of fibrino necrotic or caseous cast in the tracheal bifurcation and extending in the bronchi and were pathognomonic of respiratory distress. The lung lesions were mild to marked oedema and congestion. In some of flocks affected during early age air sacs revealed thickening with fibrin deposition. The liver and spleen in early infected flock showed congestion, mild petechial haemorrhages and necrotic foci as well as pale and swollen kidneys. Microscopic changes were most consistently seen in the trachea and bronchi and variable from mild to severe in nature. In many cases trachea and bronchi showed congestion along with infiltration of mononuclear cells in the mucosa and submucosa and presence of fibrino necrotic masses in the lumen. The tracheal lesions were mainly confined to mucosal glands and epithelial lining. Loss of cilia and hypertrophy of mucous gland were noticed in early stages of the disease followed by degeneration and desquamation of epithelium with infiltration of mononuclear cells leading to thickening of tracheal mucosa. The microscopic lesions observed in bronchi were fibrino necrotic masses in the bronchial lumen which were occluding the air passage. The microscopic lesions observed in lungs were vascular engorgement and haemorrhages, smooth muscle hypertrophy and infiltration of mononuclear cells in the parabronchi. Air sac lesions were marked by presence of fibrinous exudate and mononuclear cells infiltration. Molecular studies were carried out for detection of LPAI (H9N2), IB virus, ND virus and avian pathogenic E.coli (APEC) from pooled tissue samples of bronchi and trachea containing caseous plug of the twenty five broiler flocks. All the twenty five flocks (100%) were found positive for LPAI (H9N2) virus as well as for pathogenic E.coli, whereas IB virus, ND virus were detected in 4 (16%) and 8 (32%) farms respectively. In addition to above it was also seen that out of twenty-five broiler flocks, all the twenty five (100 percent) were positive for LPAI + APEC, three flocks (12 per cent) were positive LPAI + APEC + IBV; seven flocks (28 per cent) positive for LPAI + APEC + NDV; one flocks (4 per cent) positive for LPAI + APEC + IBV + NDV. To analyze the risk of secondary E. coli infection in case of respiratory distress, isolation, identification and characterization of E. coli from pooled tissue samples of caseous bronchial plugs from each flock was undertaken. Out of twenty-five tissue samples processed for bacterial isolation, all the samples (100%) were found positive for secondary E.coli infection. The antibiotic sensitivity test was carried out from all the isolates obtained from affected flock. The isolates were found highly sensitive to antibiotic disc colistin (Methane sulphonate) (100%) and meropenem (100%) followed by levofloxacin (76.67%), ceftriaxone (64%), gentamicin (60%), amikacin (60%), co-trimoxazole (40%), amoxycilin-clavunic acid (08%) and imepenem (08%). Molecular characterization of E. coli was carried out for detection of presence of four virulence genes i. e. iss, papC, tsh and vat. DNA was extracted from tissue samples (caseous plugs) and subjected to PCR with gene specific primer pairs. Out of twenty-five tissue samples, the iss gene, vat gene, tsh and papC gene found in 22 (88%), 13 (52%) , 15 (60%) and 8 (32%) in respiratory affected farm, respectively.
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    ThesisItemOpen Access
    STUDIES ON PROLIFERATION BIOMARKERS IN SQUAMOUS CELL CARCINOMA OF BULLOCK HORN BY IMMUNOHISTOCHEMISTRY AND ITS CORRELATION WITH HISTOPATHOLOGY, 2996
    (JAU, JUNAGADH, 2019-09) CHAUHAN JUHIBEN MANSUKHBHAI; D. T. FEFAR
    The present study was undertaken to evaluate the expression of proliferation biomarkers in squamous cell carcinoma of bullock horn by immunohistochemistry and to correlate with histopathological study. A total of twenty horn cancer tissue samples were collected from bullock from Teaching Veterinary Clinical Complex (TVCC) of College of Veterinary Science & A. H., JAU, Junagadh, Gujarat as well as from the field. The primary information pertaining to case viz. sex, breed, age and side of affected horn were collected. Pre-operatively, clinical observation of the horn cancer were carried out. Gross pathological lesions of cancerous growth were recorded after surgical amputation of horn. Subsequently, affected tissue samples as well as tissue of normal horn core were collected and fixed in 10% neutral-buffered formalin and later processed for histopathological confirmation of squamous cell carcinoma. Further, histopathologically confirmed SCC tissues were subjected to grading of squamous cell carcinoma based on cytoplasmic keratinization, nuclear pleomorphism, mitotic figure, mode of invasion and cellular response. Subsequently, expression study of proliferative biomarkers viz., Ki-67, PCNA and cyclin D1 were also carried out in histopathologically confirmed and graded squamous cell carcinoma as well as in normal horn core by immunohistochemistry. In present the study, the frequency of occurrence of horn cancer cases was highest in the age group of 6 to 10 years (70%) followed by 10 to 15 years (30%). Majority of the cases (70%) were recorded between 6 to 10 years of age. The breed wise frequency of horn cancer in randomly selected twenty cases under study stated that the frequency of occurrence of horn cancer was highest in Kankrej 13/20 (65%) followed by Gir 6/20 (30%) and non-descript 1/20 (5%) breeds of cattle. Clinical observations found in horn cancer cases were frequent shaking of head, tilting of head at the affected side, tilting of affected horn and increase nasal discharge from affected side in advance cases. Gross examination of surgical amputation of affected horn revealed cancerous growth in the middle and distal region of the horn core in first stage of horn cancer. Histopathological examination of affected tissue from this region revealed the well-differentiated squamous cell carcinoma in five cases while low differentiated squamous cell carcinoma revealed in one case. In second stage of horn cancer, cancerous tissue found nearly in entire horn core, that was dry, hard and dirty grey in color, and revealed the well-differentiated squamous cell carcinoma in four cases as well as low differentiated squamous cell carcinoma in two cases in histopathological examination of all the affected tissue from this region. Third stage of horn cancer showed completely filled horn core with cauliflower like cancerous growth and histopathological examination of all the affected tissue from this region revealed the well-differentiated squamous cell carcinoma in one case as well as low differentiated squamous cell carcinoma in seven cases. Histopathologically, all collected samples found positive for squamous cell carcinoma of horn core, which further classify into well-differentiated and poorly differentiated SCC. Well differentiated SCC revealed higher cytoplasmic keratinization, low nuclear differentiation, less mitotic figures, well defined border line and mild cellular response. The low cytoplasmic keratinization, high nuclear differentiation, high mitotic figures, poorly defined border line and high cellular response were found in poorly differentiated SCC. Immunohistochemically expression study of Ki-67 and PCNA revealed showed strong immunorecativity in low differentiated SCC and weak immunoreactivity in well-differentiated SCC and normal horn core tissue. The expression of Ki-67 and PCNA significantly found higher in poorly differentiated SCC compared to well-differentiated SCC. The expression study of cyclin D1 could not be appreciated in any of the squamous cell carcinoma as well as in normal horn tissue. Hence, the present study suggested that analysis of Ki-67 and PCNA expression in tumoral tissue may help to predict the clinical course and prognosis in patients with squamous cell carcinoma.
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    ThesisItemOpen Access
    STUDIES ON PROLIFERATION BIOMARKERS IN SQUAMOUS CELL CARCINOMA OF BULLOCK HORN BY IMMUNOHISTOCHEMISTRY AND ITS CORRELATION WITH HISTOPATHOLOGY 2996
    (JAU, JUNAGADH, 2019-09) CHAUHAN JUHIBEN MANSUKHBHAI; D. T. FEFAR
    The present study was undertaken to evaluate the expression of proliferation biomarkers in squamous cell carcinoma of bullock horn by immunohistochemistry and to correlate with histopathological study. A total of twenty horn cancer tissue samples were collected from bullock from Teaching Veterinary Clinical Complex (TVCC) of College of Veterinary Science & A. H., JAU, Junagadh, Gujarat as well as from the field. The primary information pertaining to case viz. sex, breed, age and side of affected horn were collected. Pre-operatively, clinical observation of the horn cancer were carried out. Gross pathological lesions of cancerous growth were recorded after surgical amputation of horn. Subsequently, affected tissue samples as well as tissue of normal horn core were collected and fixed in 10% neutral-buffered formalin and later processed for histopathological confirmation of squamous cell carcinoma. Further, histopathologically confirmed SCC tissues were subjected to grading of squamous cell carcinoma based on cytoplasmic keratinization, nuclear pleomorphism, mitotic figure, mode of invasion and cellular response. Subsequently, expression study of proliferative biomarkers viz., Ki-67, PCNA and cyclin D1 were also carried out in histopathologically confirmed and graded squamous cell carcinoma as well as in normal horn core by immunohistochemistry. In present the study, the frequency of occurrence of horn cancer cases was highest in the age group of 6 to 10 years (70%) followed by 10 to 15 years (30%). Majority of the cases (70%) were recorded between 6 to 10 years of age. The breed wise frequency of horn cancer in randomly selected twenty cases under study stated that the frequency of occurrence of horn cancer was highest in Kankrej 13/20 (65%) followed by Gir 6/20 (30%) and non-descript 1/20 (5%) breeds of cattle. Clinical observations found in horn cancer cases were frequent shaking of head, tilting of head at the affected side, tilting of affected horn and increase nasal discharge from affected side in advance cases. Gross examination of surgical amputation of affected horn revealed cancerous growth in the middle and distal region of the horn core in first stage of horn cancer. Histopathological examination of affected tissue from this region revealed the well-differentiated squamous cell carcinoma in five cases while low differentiated squamous cell carcinoma revealed in one case. In second stage of horn cancer, cancerous tissue found nearly in entire horn core, that was dry, hard and dirty grey in color, and revealed the well-differentiated squamous cell carcinoma in four cases as well as low differentiated squamous cell carcinoma in two cases in histopathological examination of all the affected tissue from this region. Third stage of horn cancer showed completely filled horn core with cauliflower like cancerous growth and histopathological examination of all the affected tissue from this region revealed the well-differentiated squamous cell carcinoma in one case as well as low differentiated squamous cell carcinoma in seven cases. Histopathologically, all collected samples found positive for squamous cell carcinoma of horn core, which further classify into well-differentiated and poorly differentiated SCC. Well differentiated SCC revealed higher cytoplasmic keratinization, low nuclear differentiation, less mitotic figures, well defined border line and mild cellular response. The low cytoplasmic keratinization, high nuclear differentiation, high mitotic figures, poorly defined border line and high cellular response were found in poorly differentiated SCC. Immunohistochemically expression study of Ki-67 and PCNA revealed showed strong immunorecativity in low differentiated SCC and weak immunoreactivity in well-differentiated SCC and normal horn core tissue. The expression of Ki-67 and PCNA significantly found higher in poorly differentiated SCC compared to well-differentiated SCC. The expression study of cyclin D1 could not be appreciated in any of the squamous cell carcinoma as well as in normal horn tissue. Hence, the present study suggested that analysis of Ki-67 and PCNA expression in tumoral tissue may help to predict the clinical course and prognosis in patients with squamous cell carcinoma.
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    ThesisItemOpen Access
    STUDIES ON MAJOR RESPIRATORY PATHOGENS OF BROILER CHICKENS IN AND AROUND JUNAGADH 2824
    (JAU, JUNAGADH, 2019-06) Chhatrola Mayur H.; Dr. D. T. Fefar
    The present research work was carried out to determine the etiological agents responsible for respiratory distress with special reference to presence of LPAI (H9N2), IB virus, ND virus and E. coli infection in broiler birds. The study comprised of epidemiological information in relation to farm wise mortality and age wise mortality, gross and histopathological examination of involved respiratory organs especially trachea, bronchi, lung and air sacs, detection of LPAI (H9N2) virus, IB virus and ND virus by RT-PCR, detection of avian pathogenic E. coli by PCR and also isolation as well as antibiogram against E.coli from the birds affected with respiratory distress. Studies on the farm incidence and age wise mortality among twenty five broiler flocks having total population of 75,800 birds of second to fifth week of age showed an average mortality rate of 3.25 percent ranging between 1.33 to 8.00 percent. The week wise mortality was found highest during third week of age followed by second, fourth and fifth weeks of age. Gross pathological lesions were predominantly found in organs of respiratory system i.e. in trachea, bronchi, lungs and air sacs. The tracheal lesions observed were severe congestion, excessive mucous exudate, presence of fibrino necrotic or caseous cast in the tracheal bifurcation and extending in the bronchi and were pathognomonic of respiratory distress. The lung lesions were mild to marked oedema and congestion. In some of flocks affected during early age air sacs revealed thickening with fibrin deposition. The liver and spleen in early infected flock showed congestion, mild petechial haemorrhages and necrotic foci as well as pale and swollen kidneys. Microscopic changes were most consistently seen in the trachea and bronchi and variable from mild to severe in nature. In many cases trachea and bronchi showed congestion along with infiltration of mononuclear cells in the mucosa and submucosa and presence of fibrino necrotic masses in the lumen. The tracheal lesions were mainly confined to mucosal glands and epithelial lining. Loss of cilia and hypertrophy of mucous gland were noticed in early stages of the disease followed by degeneration and desquamation of epithelium with infiltration of mononuclear cells leading to thickening of tracheal mucosa. The microscopic lesions observed in bronchi were fibrino necrotic masses in the bronchial lumen which were occluding the air passage. The microscopic lesions observed in lungs were vascular engorgement and haemorrhages, smooth muscle hypertrophy and infiltration of mononuclear cells in the parabronchi. Air sac lesions were marked by presence of fibrinous exudate and mononuclear cells infiltration. Molecular studies were carried out for detection of LPAI (H9N2), IB virus, ND virus and avian pathogenic E.coli (APEC) from pooled tissue samples of bronchi and trachea containing caseous plug of the twenty five broiler flocks. All the twenty five flocks (100%) were found positive for LPAI (H9N2) virus as well as for pathogenic E.coli, whereas IB virus, ND virus were detected in 4 (16%) and 8 (32%) farms respectively. In addition to above it was also seen that out of twenty-five broiler flocks, all the twenty five (100 percent) were positive for LPAI + APEC, three flocks (12 per cent) were positive LPAI + APEC + IBV; seven flocks (28 per cent) positive for LPAI + APEC + NDV; one flocks (4 per cent) positive for LPAI + APEC + IBV + NDV. To analyze the risk of secondary E. coli infection in case of respiratory distress, isolation, identification and characterization of E. coli from pooled tissue samples of caseous bronchial plugs from each flock was undertaken. Out of twenty-five tissue samples processed for bacterial isolation, all the samples (100%) were found positive for secondary E.coli infection. The antibiotic sensitivity test was carried out from all the isolates obtained from affected flock. The isolates were found highly sensitive to antibiotic disc colistin (Methane sulphonate) (100%) and meropenem (100%) followed by levofloxacin (76.67%), ceftriaxone (64%), gentamicin (60%), amikacin (60%), co-trimoxazole (40%), amoxycilin-clavunic acid (08%) and imepenem (08%). Molecular characterization of E. coli was carried out for detection of presence of four virulence genes i. e. iss, papC, tsh and vat. DNA was extracted from tissue samples (caseous plugs) and subjected to PCR with gene specific primer pairs. Out of twenty-five tissue samples, the iss gene, vat gene, tsh and papC gene found in 22 (88%), 13 (52%) , 15 (60%) and 8 (32%) in respiratory affected farm, respectively.
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    ThesisItemOpen Access
    ETIO-PATHOLOGICAL AND PROINFLAMMATORY CYTOKINE EXPRESSION STUDIES ON ENDOMETRITIS IN BUFFALOES 2512
    (JAU,JUNAGADH, 2018-04) AMIT R. BHADANIYA; DR. M.C. PRASAD
    The present inestigation was carried out to study the prevalence of endometritis, its bacterial etiology / their antibiotic sensitivity, pro-inflamatory cytokine profiling /their role in understanding the pathogenesis of endometritis leading to early diagnosis and pathomorphological changes on slaughter house materials collected from slaughter house at Junagadh. The buffaloes were comparatively aged ones and a total of 110 genitalia were collected and processed in Pathology Department for fulfilling various objectives mentioned above. The prevalence of endometritis was recorded in 81% (89/110) slaughtered buffaloes and it was also categorized as acute (25.45%), subacute (20.90%) and chronic (34.55%) based on pathomorphological feature. Acute endometritis was characterized by severe congestion along with marked stromal edema, degenerative changes and focal denudation of luminal epithelium, focal hemorrhage in sub epithelial zone, infiltration of inflammatory cells predominantly polymorphonuclear cells and mononuclear cells in lamina propria, infiltration of mononuclear cells in glandular lumina and peri-endometrial glands. Subacute endometritis consisted of denudation of luminal epithelium, congestion, stromal edema and focal haemorrhagic spot, infiltration of mononuclear cells in lamina propria, glandular lumina and around the atrophied endometrial gland, glandular dilation, hyperplasia of mucosal epithelium, atrophy of endometrial glands and thickening of blood vessels. The main features of chronic endometritis were desquamation of mucosal epithelium, infiltration of mononuclear cells and plasma cells in sub epithelial zone, dilatation of endometrial glands with degenerative changes, infiltration of mononuclear cells in glandular lumina and periglandular region with narrowing of glandular lumina, perivascular and periglandular fibrosis leading to severe thickening of blood vessels resulting in narrowing of their lumina and transformation of endometrial epithelial cells into low cuboidal against the normal columnar epithelium. Beside the above histopathological lesions three uterine samples revealed adenomyosis and twenty two genitalia showed metritis. The bacterial cultural examination was carried out on all the110 uterine swabs collected from slaughtered buffaloes. Fifty six (50.90%) uterine samples showed the growth of various bacteria and remaining 54 swabs (49.10%) were found to be sterile. Out of the 56 uterine samples 50 swabs (89.28%) showed single isolate and remaining 06 (10.72%) exhibited mixed infection. Bacterial isolates comprised of Staphylococcus spp. (11.82%), Escherichia coli (8.18%), Micrococcus spp. (8.18%), Corynebacterium spp. (7.27%), Fusobacterium spp. (4.55%), Pseudomonas spp. Abstract… (2.73%), Bacillus spp. (1.82%), Streptococcus spp. (0.91%) and mixed infections consisted of two different types of isolates in different combinations (E. coli + Corynebacterium spp. – 2.73%, E. coli + Staphylococcus spp. – 1.82% and E. coli + Bacillus spp. – 0.91%). The Escherichia coli and Staphylococcus spp. isolates (24.19%) topped the list followed by Corynebacterium spp. (17.74%), Micrococcus spp. (14.52%), Fusobacterium spp. (8.06%), Pseudomonas spp. (4.84%), Bacillus spp. (4.84%) and Streptococcus spp. (1.61%). Overall antimicrobial susceptibility of 62 bacterial isolates showed highest sensitivity against Chloramphenicol (83.9%) followed by Gentamicin (80.6%), Levofloxacin (77.4%), Oxytetracycline (77.4%), Ceftriaxone/Sulbactam (69.3%), Cefoperazone/Sulbactam (61.2%) and Amoxicillin/Sulbactam (33.9%). Subclinical and clinical endometritis were adjudged on the basis of endometrial cytology and white side test for cytokine profiling. The endometrial cytology showed more than 5% polymorphonuclear cells (PMNs) among 200 cell counted in 45 (40.90%) cases out of 110 sample. Eighteen genitalia out of 110 genitalia having mucopurulent or purulent discharge and positive on white side test were taken as clinical endometritis. A total of 36 categorized genitalia were divided into three groups namely, Group-I (n=12; Normal), Group-II (n=12; Clinical Endometritis / CE) and Group-III (n=12; Subclinical Endometritis / SCE). Further, each group was subdivided as follicular (n=6) and luteal (n=6) stages of the ovarian cycle based on gross morphology of ovaries. The expression of CD14, TLR-4 and IL-1β encoding gene in endometrium of buffaloes with subclinical endometritis were 1.87, 2.71 and 3.48 fold, while samples with clinical endometritis showed only 2.90, 4.76 and 8.02 fold for CD14, TLR-4 and IL-1β, respectively, though numerical value of expression was higher in subclinical and clinical endometritis but it was at par with control. The expression profile for IL- 6, IL-8 and TNF-α mRNA for subclinical endometritis were 11.95, 14.87 and 12.95 fold while for clinical endometritis, values were 18.17, 37.97 and 28.83 fold for IL-6, IL-8 and TNF-α mRNA, respectively, which were significantly higher in subclinical and clinical endometritis as compared to apparently normal samples at follicular phase. In the luteal group, the subclinical endometritis sample showed 2.13, 6.43, 14.52, 22.38, 18.64 and 2.82 fold respectively, for CD14, IL-1β IL-6, IL-8, TNF-α and TLR-4 mRNA. Except TLR-4, the values of expression were significantly higher when compared with apparently normal. Whereas, the expression values of clinical endometritic uteri were 4.01, 7.15, 17.20, 45.55, 52.58 and 32.95 fold respectively, for CD14, TLR-4, IL-1β, IL-6, IL-8 and TNF-α mRNA which were significantly higher as compared to apparently normal uteri. Based on present study it was concluded that: i) the prevalence of endometritis in slaughtered buffalo was 81% and most common bacteria involved were E coli and Staphylococcus which were highly sensitive to Chloramphenicol, ii) the mRNA expression values of different cytokines (CD14, IL-1β, IL-6, IL-8 and TNF-α) might represent a potential marker gene to detect endometritis in buffaloes, iii) chronic endometritis was the major problems seen in slaughtered buffaloes and as these animals cannot reproduce, caused economic loss to livestock owners / farmers and this is why such animals are sold for slaughtering purposes.