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Open Access
EXPRESSION OF MECHANISTIC TARGET OF RAPAMYCIN(mTOR) AND DISHEVELLED, EGL 10 AND PLECKSTRIN DOMAIN CONTAINING MTOR INTERACTING PROTEIN (DEPTOR) IN CANINE SUPERFICIAL AND MAMMARY TUMOURS
(COLLEGE OF VETERINARY AND ANIMAL SCIENCES MANNUTHY, THRISSUR, 2019-09-30) RAIMON MATHEW; Sajitha I.S.
The present work was undertaken to study the expression of two proteins, critical in the cell signaling pathway namely, mechanistic target of Rapamycin (mTOR) and Dishevelled, EGL10 and Pleckstrin domain containing mTOR interacting protein (DEPTOR) in canine mammary and superficial tumours. Excision biopsy samples of mammary and skin tumour suspected growths from dogs presented to University Veterinary hospitals, Mannuthy and Kokkalai were collected for the study during the period from January 2018 to March 2019. Out of the 129 tumour suspected cases, 22 were mammary tumour growths and 17 were superficial tumour growths. Age wise analysis revealed that the mean age of the dogs affected with mammary tumours and superficial tumours was 7.85±0.41 years and 6.55±0.59 years respectively. The highest incidence of both the tumours was seen in the age group of seven to nine years. All the mammary tumours were recorded in female dogs but in case of superficial tumours male dogs (58.82 per cent) were more affected. Data on breed-wise occurrence showed the highest occurence of both the type of tumours in Labradors. Among the mammary tumours, 90 per cent were malignant and 10 per cent were benign, where as in case of superficial tumours majority (76.47 per cent) were benign in nature. Grossly CMTs varied in shape, colour and consistency with a mean size of 6.70±0.60 cm. The superficial tumours were predominantly round to oval in shape with grayish white colour having a mean size of 4.65±0.37cm. On grading of malignant CMTs, majority of the simple carcinomas, which are the most malignant type, had higher grade of II or III. Immunohistochemistry for mTOR and DEPTOR has revealed that both the proteins were significantly expressed (p0.05) with expression of these proteins. The mean expression of mTOR in mammary tumours and superficial tumours was significantly higher (p<0.01) than that of DEPTOR. Further, it was also found that there was significant positive correlation (p<0.01) between the expression of mTOR and DEPTOR in both these type of tumours. Thus, the present study identified that the two key proteins in cell signaling pathway, the mTOR and DEPTOR, were significantly expressed in canine mammary and superficial tumours. Further it was shown that their expression was positively correlated. Hence the oncogene role of DEPTOR in CMTs and CSTs was established through the study.
Open Access
EVALUATION OF IN VIVO HOST RESPONSE OF DIFFERENTLY DECELLULARISED OMENTAL SCAFFOLDS
(COLLEGE OF VETERINARY AND ANIMAL SCIENCES MANNUTHY, THRISSUR, 2019-12-31) ASHNA SULAIMAN; Dhanush Krishna B.
In this study, efficiency of decellularisation protocols as well as host response of differently decellularised bovine omental scaffolds were evaluated in a rat model. Bile treated omental scaffold (BBO), triton X- 100 treated omental scaffolds (TBO) and un-processed omental scaffolds (UBO) were used during the implantation period along with sham control (SCO). Efficiency of different decellularisation protocols was assessed by DNA quantification, 4, 6- diamidino-2-phenylindole (DAPI) staining, scanning electron microscopy and histological examination. Cellular and tissue response was evaluated semi quantitatively using ISO-10993-6: 2016 at 7, 14 and 30 days post- implantation (PI). CD4+ and CD8+ T-cells in retrieved implants and spleen were evaluated immunohistochemically to study the graft acceptance of these implants. Proliferation index was also evaluated by Ki67 immunolabelling. The results of this study showed that bile treatment had better decellularisation efficiency than triton X- 100. BBO showed better remodeling response in terms of neovascularisation, fatty infiltration and neocollagenisation than TBO and UBO. At all time periods studied, BBO implant had significantly (p< 0.05) lesser CD4+ per cent and higher CD8+ per cent than TBO and UBO, respectively. UBO showed significantly higher CD4+ per cent at all time periods than BBO and TBO. The CD4+/CD8+ ratio for BBO was significantly (p< 0.05) lower than TBO and UBO. Besides, BBO showed high Ki67 index (p<0.05) than TBO at 30 days PI. These results document that bile as an effective decellularising agent with less tissue toxicity than chemical detergent, triton X- 100. Apart from these, BBO enhanced remodeling potential through neovascularisation, cellular proliferation and neocollagenisation with significantly lower CD4+/CD8+ ratio and higher Ki67 index. Thus, bile treated omental scaffold may be a promising candidate for applications in human and veterinary medicine for improving patient’s life.
Open Access
EVALUATION OF IN VIVO HOST RESPONSE OF DIFFERENTLY DECELLULARISED OMENTAL SCAFFOLDS
(COLLEGE OF VETERINARY AND ANIMAL SCIENCES MANNUTHY, THRISSUR, 2019-08-29) SULAIMAN, ASHNA; Krishna B., Dhanush
In this study, efficiency of decellularisation protocols as well as host response of differently decellularised bovine omental scaffolds were evaluated in a rat model. Bile treated omental scaffold (BBO), triton X- 100 treated omental scaffolds (TBO) and un-processed omental scaffolds (UBO) were used during the implantation period along with sham control (SCO). Efficiency of different decellularisation protocols was assessed by DNA quantification, 4, 6- diamidino-2-phenylindole (DAPI) staining, scanning electron microscopy and histological examination. Cellular and tissue response was evaluated semi- quantitatively using ISO-10993-6: 2016 at 7, 14 and 30 days post- implantation (PI). CD4+ and CD8+ T-cells in retrieved implants and spleen were evaluated immunohistochemically to study the graft acceptance of these implants. Proliferation index was also evaluated by Ki67 immunolabelling. The results of this study showed that bile treatment had better decellularisation efficiency than triton X- 100. BBO showed better remodeling response in terms of neovascularisation, fatty infiltration and neocollagenisation than TBO and UBO. At all time periods studied, BBO implant had significantly (p< 0.05) lesser CD4+ per cent and higher CD8+ per cent than TBO and UBO, respectively. UBO showed significantly higher CD4+ per cent at all time periods than BBO and TBO. The CD4+/CD8+ ratio for BBO was significantly (p< 0.05) lower than TBO and UBO. Besides, BBO showed high Ki67 index (p<0.05) than TBO at 30 days PI. These results document that bile as an effective decellularising agent with less tissue toxicity than chemical detergent, triton X- 100. Apart from these, BBO enhanced remodeling potential through neovascularisation, cellular proliferation and neocollagenisation with significantly lower CD4+/CD8+ ratio and higher Ki67 index. Thus, bile treated omental scaffold may be a promising candidate for applications in human and veterinary medicine for improving patient’s life.
Open Access
IMMUNOHISTOCHEMICAL EXPRESSION OF EPIDERMAL GROWTH RECEPTOR AND PATHOLOGY OF ORAL AND PHARYNGEAL NEOPLASMS IN DOGS
(COLLEGE OF VETERINARY AND ANIMAL SCIENCES, POOKODE WAYANAD, 2019-10-03) NAIK, MADHURA PRASHANT; R., Anoopraj
The present study entitled “Immunohistochemical expression of epidermal growth factor receptor and pathology of oral and pharyngeal neoplasms in dogs” was conducted for obtaining the data on the occurrence of oral and pharyngeal neoplasms in dogs, classification and grading and to compare the expression of epidermal growth factor receptor expression in tumour and normal tissues. A total of 23 oral and pharyngeal lesions were recorded. Out of the 23 lesions, 20 cases were diagnosed as neoplasms and 3 cases were of non-neoplastic origin. Age groups of seven to nine years showed highest incidence of the tumour (45 per cent), 85 per cent of the affected animals were males, while Labrador was the most affected breed (35 per cent). Gingiva formed the common site affected with tumours (60 per cent). The occurrence of malignant and mesenchymal tumours was 55 per cent and 80 per cent respectively. The malignant tumours encountered were histiocytic sarcomas, squamous cell carcinoma, fibrosarcoma, hemangiosarcoma, telangiectatic osteosarcoma and melanoma. Benign tumours included peripheral odontogenic fibroma, osteoma, oral papilloma and fibroma. Malignant tumours were graded based on specific histopathological parameters. Three cases of gingival hyperplasia were also studied. Expression of epidermal growth factor receptor (EGFR) was assessed in tumour specimens and the corresponding normal tissues by immunohistochemistry. Archived samples such as basal cell carcinoma, squamous cell carcinoma, acanthomatous ameloblastoma, desmoplastic ameloblastoma, papilloma and melanoma were also included in the study. When compared to normal tissues, significant strong over expression of EGFR was observed in squamous cell carcinoma, basal cell carcinoma, histiocytic neoplasms, hemangiosarcoma and melanoma while fibrosarcoma and ameloblastoma showed moderate over expression. No significant difference in EGFR expression was observed in papilloma and bone tumours.
Open Access
PATHOLOGICAL STUDIES AND MOLECULAR DETECTION OF Neospora caninum, Ehrlichia canis and Borrelia burgdorferi INFECTION IN DOGS WITH NERVOUS DISORDERS
(COLLEGE OF VETERINARY AND ANIMAL SCIENCES, POOKODE WAYANAD, 2019-10-03) H. B, VIDYARANI.; K. S, Prasanna
Neurological disorders in dogs with brain affections have multiple aetiology and often confused as rabies. In the present research work we studied gross and histopathological lesions in the rabies negative brain samples and screened them for the presence of Borrelia burgdorferi (B. burgdorferi), Ehrlichia canis (E. canis) and Neospora caninum (N. caninum). Out of the 84 dog carcasses presented to the Department of Veterinary Pathology, College of Veterinary and Animal Sciences, Pookode during January 2018 to June 2019, 36 brain samples were found negative for rabies (42.9 per cent) by using direct Fluorescent Antibody test. The prominent histopathological lesions in the cerebrum, cerebellum and meninges were classified as vascular (86 per cent) and cellular changes (41.66 per cent). The vascular changes were congestion, haemorrhage and vasculitis (90. 3 per cent) and perivascular cuffing with mononuclear cells infiltration (22.6 per cent). The prominent neuronal changes were chromatolysis, shrinkage of neurons, perineuronal vacuolations, degeneration of neurons and neuronophagia. The common glial changes observed were satellitosis, peri glial vacuolations and gliosis. We did other diagnostic methods of blood smear examination for detection of morulae of E. canis and rapid immunochromatographic test for detection of antibodies against the B. burgdorferi in blood samples. Faecal sample examination was carried out for detection of N. caninum oocysts. Stained brain and tissue impression smears (Sellers and Giemsa stain) were examined for the detection of parasitic stages of N. caninum. All these tests were found negative for any of the specific pathogen targeted. Molecular tests were carried out for B. burgdorferi detection by nested PCR targeting 16SrRNA gene and conventional PCR for N. caninum detection by targeting Nc5 gene and none of the samples were found positive. For E. canis detection, the conventional PCR targeting 16SrRNA gene was carried out and one sample (2.78 per cent) was found positive for both genus and species-specific primer sets used. The specific lesions in the brain sample positive for E. canis were congestion, haemorrhage, diffuse vasculitis and perivascular cuffing with mononuclear cell infiltration in cerebrum and cerebellum.
Open Access
PATHOLOGICAL STUDIES OF ENTERO-HEPATIC AFFECTIONS IN DUCKS AND SCREENING FOR SPECIFIC VIRAL ETIOLOGY
(COLLEGE OF VETERINARY AND ANIMAL SCIENCES, POOKODE WAYANAD, 2019-10-03) SANJAY RAO, NIKHIL; M., Pradeep
Duck production holds an important position in the Indian poultry scenario, being the second most domesticated species of poultry in the country after chicken. Ducks are reared for both meat and egg production and a healthy liver and intestine capacitate the bird for its best performance and help the farmers for better profitability. Reports on systematic studies on the liver and intestinal lesions of ducks are scanty in India or have only been studied in relation to particular disease aetiology. Even though ducks show less susceptibility to diseases compared to the chicken, a few infectious and non-infectious diseases that affects liver and intestine were reported in ducks from many duck rearing countries. Among the viral diseases, the most important infections reported are duck enteritis virus (DEV) and duck virus hepatitis (DVH). Duck virus hepatitis (DVH) is an acute and contagious disease affecting liver but so far has not been reported in India. The present study was aimed at systematic post mortem recording of gross and histopathological lesions in the liver and intestine of duck carcasses and screening for DVH viruses and DEV by Polymerase chain reaction (PCR) using species specific primers. A total of 112 duck carcasses were examined in the study. Liver showed enlargement with characteristic deep yellow discoloration and firmness in 43.75 per cent cases. Pale yellow or dark brown friable liver recorded in 21.43 per cent cases. Amyloidosis of varying degrees were observed in 61.61 per cent cases of duck livers examined. A correlation between the severity of amyloidosis and gross lesions could also be derived. The enlarged, yellow and firm liver was observed to have moderate to severe amyloidosis histologically, while pale pink liver had moderate amyloidosis. The intestine also showed amyloid deposits in such cases without appreciable gross lesions. Ducks farmed under intensive system of rearing had a higher incidence of amyloidosis particularly in White Pekin ducks. Friable consistency of liver was either due to severe congestion or fatty change. Hepatocellular carcinoma with cirrhosis was identified in one case. DVE and DHV were not detected in any of the cases. Further screening for DVH viruses in a larger population of ducks is warranted. The absence of DEV in the current study may be due to effective control of the disease through appropriate vaccination strategies. The high incidence of amyloidosis in this study and the possible risk of dietary consumption of amyloid livers and its zoonotic potential needs to be investigated.
Open Access
HISTOPATHOLOGICAL AND IMMUNOHISTOCHEMICAL STUDIES OF INFECTIOUS BURSAL DISEASE IN CHICKS
(College of Veterinary and animal Science,Mannuthy, 2019) RAHMAAN S.B.; Sajitha I. S.
The study was conducted to detect the presence of Infectious Bursal Disease (IBD) virus in the bursa of Fabricius of unvaccinated chicks below three weeks of age by polymerase chain reaction (PCR), study the pathology induced by the virus in the lymphoid organs of affected chicks and to evaluate the lymphocyte subpopulation in the bursa of affected chicks using immunohistochemistry (IHC). One hundred samples were collected from dead birds brought for postmortem examination to the Department of Veterinary Pathology, College of Veterinary and Animal Sciences, Mannuthy as well as from egger nurseries and private farms in and around Thrissur. The detailed history and gross lesions were recorded and tissue samples such as bursa of Fabricius, spleen, thymus and other organs were collected for histopathology and PCR. Out of one hundred samples, eight samples were positive for IBD virus by PCR. Gross lesions such as splenomegaly, haemorrhagic typhilitis, swollen and congested kidneys were the common findings in the affected birds. Histopathologically, lymphoid depletion and infiltration of heterophils in bursa of Fabricius, moderate to diffuse lymphoid depletion in spleen and decrease in thymocyte density with congestion in thymus were noticed. Viral antigen was demonstrated in bursa, spleen and liver of affected birds and influx of CD4+ and CD8+ lymphocytes were seen in the bursa of affected chicks by IHC. The present study confirms the occurrence of subclinical IBD in poultry in and around Thrissur and the organ of choice for demonstrating IBD viral antigen by IHC was found to be bursa of Fabricius when compared to other lymphoid organs.
Open Access
MOLECULAR DIAGNOSIS AND PATHOLOGY OF PORCINE REPRODUCTIVE AND RESPIRATORY SYNDROME VIRUS AND PORCINE PARVO VIRUS INFECTIONS
(College of Veterinary and animal Science,Mannuthy, 2019) SAIRAM R; Dhanush Krishna B.
Porcine reproductive and respiratory syndrome virus (PRRSV) and porcine parvo virus (PPV) are important emerging viral diseases having significant impact on global swine industries. Hence, the present research work was designed with the objective of detecting PRRSV and PPV from naturally occurring cases in swine population. Gross and histopathological lesions due to PRRSV and PPV infections were studied and these infections were further confirmed with immuno-histochemistry (IHC). 56 samples from pig carcasses suspected for PRRSV and PPV infections were collected during the study period from June 2018 to June 2019 from different swine farms across Kerala. Reverse transcriptase polymerase chain reaction (RT-PCR) and polymerase chain reaction (PCR) identified eight cases positive for PRRSV and one case positive for PPV, respectively. Common gross lesions of PRRSV positive cases were non-collapsed voluminous lungs, enlarged and congested lymph nodes, splenomegaly, hemopericardium with multifocal whitish areas of necrotic spots in the liver. Gross lesions in PPV positive cases were non-collapsed voluminous lungs, fibrinous pericarditis, splenomegaly and multifocal whitish necrotic foci in kidneys. Histopathologically, PRRSV cases revealed interstitial pneumonia and severe lymphoid depletion in the tonsils of the soft palate, spleen and lymph nodes. PPV positive case displayed interstitial pneumonia, moderate lymphoid depletion in various lymphoid organs and interstitial nephritis. With IHC, viral antigens in positive cases of PRRSV and PPV were demonstrated in lungs and lymphoid organs. In addition, phylogenetic analysis of PRRSV isolate in this study was genetically related to other Indian isolates but forms a separate clade whereas PPV showed close relationship with Korean and Pookode strains. Therefore, this study establishes the presence of PRRSV and PPV in the swine population of Kerala. Routine screening of these viruses in swine farms needs to be practised as an effective measure to control these diseases. So, this study will serve as an evidence for the importance of vaccination against PRRSV and PPV infections of swine populations in near future.
Open Access
PROTECTIVE EFFECT OF AEGLE MARMELOS (KOOVALAM) AND ANDROGRAPHIS PANICULATA (KIRIYATHA) IN SUBLETHAL AFLATOXICOSIS OF BROILER CHICKEN
(College of Veterinary and animal Science,Mannuthy, 2018) ANEESH A; Ajith Jacob George
The present study was undertaken to assess the protective effect of Aegle marmelos (“koovalam”) and Andrographis paniculata (“kiriyatha”) in sublethal aflatoxicosis of broiler chicken. Protective effect of Aegle marmelos and Andrographis paniculata, individually and in combination was checked. Aflatoxin (AF) was produced using Aspergillus flavus NRRL 6513 culture. The pooled maize culture powder was estimated to contain 63.77 ppm of AF through thin layer chromatography method. The Aegle marmelos mature leaves powder and Andrographis paniculata whole plant powder were analysed to detect various active chemical constituents using phytochemical screening. Forty-eight numbers of Vencobb 400 strain, dayold broiler chicks were randomly divided into eight groups of six birds each including control with standard feed (T1), AF control containing standard feed and 100 ppb AF (T2), standard feed with 0.1 per cent A. marmelos powder and 0.1 per cent A. paniculata powder (T3), standard feed with 100ppb AF and 0.10 per cent A. marmelos powder (T4), standard feed with 100ppb AF and 0.10 per cent A. Paniculata powder (T5), standard feed with 100 ppb AF and 0.20 per cent A. marmelos powder (T6), standard feed with 100 ppb AF and 0.20 per cent A. paniculata powder (T7) and standard feed containing 100 ppb AF 0.1 per cent A. marmelos powder and 0.1 per cent A. paniculata powder (T8). Blood was collected from the wing vein of all the birds at day 7 and 14 for enzymology and on day 21 for heamatological and biochemical analysis. All birds were sacrificed at the end of 21 days and detailed post mortem was conducted. Phytochemical screening detected presence of alkaloids, tannins and flavonoids in A. marmelos leaves and alkaloids, tannins, flavonoids and triterpenes in A. paniculata. Neither mortality nor clinical signs could be observed among any of the treatment groups. Body weight and body weight gain were significantly decreased among T2 group fed with 100 ppb of AF. Addition of Andrographis paniculata and Aegle marmelos powders at concentrations of 0.1 per cent, 0.2 per cent and in combination at 0.1 per cent each along with AF resulted in similar body weights as that of control group. Feed conversion ratio was significantly increased among T2 group compared to all the other treatments. Andrographis paniculata and Aegle marmelos were able to alleviate negative effects of 100 ppb AF on relative weight of spleen. Haemoglobin concentration, volume of packed red cells and total leukocyte count were significantly reduced among T2 group of broilers. These haematological parameters were similar to that of control in birds of groups T3, T4, T5, T6, T7 and T8. Total protein and albumin values were significantly elevated in T2 group. Globulin levels and albumin:globulin ratio did not show any statistical significance between groups. Significantly increased aspartate aminotransferase and alanine aminotransferase values were observed among T2 group, but was similar to control among T3,T4,T5,T6,T7 and T8 indicating the hepatoprotective properties of both the plants. Statically significant elevation of blood urea nitrogen and creatinine was noticed in T2 group and was similar to control in other groups indicating the nephroprotective effects of both plant materials. Lipid peroxidation levels were significantly high among T2 group. Both plant materials addition in diet could bring out he lipid peroxidation levels to that of control due to their anti-oxidant properties. Pale yellowish liver was observed among T2 group broilers, all other organs were similar in gross appearance between groups. Histopathological lesions like presence of vacuolations, hyperplasia of kupffer cells of liver, mononuclear infiltration of liver, kidney and lymphoid depletion of lymphoid organs were observed in T2 group. Severity of histopathological lesions were significantly reduced with supplementation of plant materials.

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