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Open Access
EFFECTS OF ROOT EXTRACT OF SAUSSUREA LAPPA ON CHEMICAL INDUCED LIVER DAMAGE IN RATS
(CSKHPKV Palampur, 2020-09-10) ABHISHEK, KUMAR; Asrani, R.K.
Open Access
STUDIES ON PATHOLOGY OF RESPIRATORY DISEASES IN BOVINE CALVES WITH SPECIAL EMPHASIS ON BOVINE RESPIRATORY SYNCYTIAL VIRUS AND PARAINFLUENZA-3 VIRUS INFECTION.
(CSK HPKV, Palampur, 2019-07-23) Sameeksha; Gupta, V.K.
The present study was conducted to determine the prevalence and the factors responsible for respiratory diseases in bovine calves during April, 2018 to May, 2019. A total 210 nasal swabs were collected from the necropsy (18), clinically sick (130) and apparently healthy calves (62). Besides, 24 tracheal and 12 lung swabs from necropsy cases were also collected. 33 calves were examined at necropsy. Pneumonia was recorded in all 33(100%) calves and was considered an important contributor to death. Out of 33 cases, gross lesions suggestive of bronchopneumonia seen in 7/33 (21.2%), interstitial pneumonia in 4/33 (12.1%) and bronchointerstitial pneumonia in 22 (66.7%). Bronchointerstitial pneumonia revealed mixed characteristic features of suppurative bronchopneumonia and interstitial pneumonia. Staphylococcus aureus was detected in 30.3 % of the clinical samples. E. coli comprised 23.6% of the isolates in necropsy. Bacterial isolates seen in nasal tract were different from the bacterial isolates obtained from lungs / trachea. Single bacterial infection was detected in 51.5% (17/33) cases and polymicrobial infection in 45.5% (15/33) cases. Total 41 lung tissues from necropsy and 104 nasal samples from clinical cases were screened for BRSV using RT-PCR with glycoprotein gene (246bp) primers and showed overall prevalence of 50.4%. BRSV infected lung showed consolidation at the cranioventral and fibrinous pleuritis with range of changes in airways histopathologically multinucleated syncytial cells was seen. Total of 145 cases were screened for BPIV-3 using RT-PCR, no sample was found positive for primer having HN gene. In necropsy, most frequent E.coli pathotype categories were found to be enterotoxigenic (80%) & enteroaggregative (80%). In clinically sick cases frequent pathotype categories found were enterotoxigenic (63.6%). Four (33.3%) of Citrobacter freundii isolates were pathogenic with shiga toxin on PCR analysis. 17.9% Staphylococcus aureus and 15.4% Staphylococcus epidermidis were recorded as pathogenic. Male and female calves showed 20/33 (60.7%) and 13/33 (39.4%) mortalities respectively. Maximum mortality was recorded in <1 M (42.2%) old calves. Most number of deaths were recorded during the summer (36.4%) season. Combination of molecular detection (PCR) along with clinical, gross & histopathology helped in better confirmatory diagnosis of BRSV infection in calves. The present study concluded co-infection of bacterial and BRSV in respiratory infection of young calves.
Open Access
STUDIES ON THE PATHOLOGY OF NEOPLASM IN DOMESTIC ANIMALS.
(CSKHPKV, Palampur, 2019-07) Masand, Rupali; Patil, R.D.
The present work was conducted to study the pathomorphological changes of animal neoplasms by using cytological, gross and histopathological, histochemical and immunohistochemical studies. A total of ninety-six (96) tumor cases were collected during September, 2017 to June, 2019. The incidence of neoplasms was highest in canines (71.87%) followed by the bovines (16.67%), rabbits (7.2%), equines (2.08%) and caprine (1.04%). The maximum incidence of neoplasms occurred in the age group of 3-6 years (28.12%) and the incidence was highest during summer (31.25%) followed by winter (25%), monsoon (23.45%) and spring (19%) season. Benign tumors (53.12%) were higher than malignant tumors (46.87%). Location wise highest occurrence of neoplasms was recorded in mammary gland (20.33%) followed by oral region (18.75%), genitalia (11.45%). On basis of cytology, tumors of epithelial origin were 58.04%, mesenchymal origin 32.25% and round cell tumor 9.6%. Canine mammary tumors (18.84%) were found to be most frequently recorded tumors followed by squamous cell carcinoma (13.04%). Histopathological examination of papilloma, squamous cell carcinoma, Fibroma, fibrosarcoma, haemangioma, haemangiosarcoma, mast cell tumor, melanocytoma, malignant melanoma, sebaceous adenoma, mammary adenoma and adenocarcinoma, mixed mammary tumor, solid carcinoma of mammary gland, hepatocellular carcinoma, cholangiocellular carcinoma, acinar cell carcinoma, malignant renal cell carcinoma, Sertoli cell tumor, cystic papillary carcinoma revealed charecteristic benign and malignant features. Histochemical staining such as Masson’s trichrome, Van Gieson’s, Toluidine blue and Schmorl's confirmed the tumors as fibroma or fibrosarcoma, leiomyoma, mast cell tumors and melanocytoma or malignant melanoma, respectively. Silver stained nucleolar organizer region (AgNOR)count showed significantly higher count in malignant than benign tumors. Amongst malignant tumor, the highest AgNOR count was found in cholangiocellular carcinoma (6.89±2.00) followed by fibrosarcoma of mammary gland (6.64±2.47) and least in haemagiosarcoma (2.96±1.25). Immunohistochemical studies in tumor for the expression of proliferating cell nuclear antigen (PCNA), p53 and H-ras found to have diagnostic value and supported the hisopathological observations of tumors in animals. Further, TdT mediated dUTP nick end labeling (TUNEL) assay was found to be useful for determining the role of apoptosis in different animal neoplasms.
Open Access
INVESTIGATION ON PROTECTIVE EFFECT OF MEDICINAL PLANT(S) AGAINST HEPATIC DAMAGE IN RATS
(CSKHPKV, Palampur, 2019-02-08) Sharma, Rakshita; Asrani, R.K.
The present study was planned with a view to observe the In-vitro cytotoxicity of 70% ethanolic extract of plant Saussurea lappa, Phyllanthus urinaria, Rheum webbianum, Artemisia nilagirica and Zingiber crysanthum on A549, KB and SiHA cell lines. Extracts were screened for the presence of various polyphenols and main bioactive compounds of respective plant. The plant extract from 3 plants namely Phyllanthus urinaria, Saussurea lappa and Rheum webbianum was used in combination (1:2:2) to study the growth response, mortality, clinical signs, biochemical and pathological changes against DEN induced hepatic damage in rats. Acute toxicity was studied in 6 rats. For final experimentation, 42 rats were randomly divided into 6 groups, Group-1: plain control, Group-2: N-diethylnitrosamine (DEN) alone. Group-3: DEN with Silymarin @25mg, Group-4: DEN with combination of plant extracts @100mg/kg, Group-5: DEN with combination of plant extract @250mg, Group-6: DEN with combination of plant extract @500mg. The N-diethylnitrosamine (DEN) was given in drinking water to rats throughout the experiment. The route of plant extract and standard drug was oral intubation throughout the period of experimentation of 17 weeks. Results of the present study showed that rats of group-2 (DEN only) exhibited clinical signs of decrease in body weight, hair loss, dullness, and depression. ALT, AST, Creatinine activity was found to increase, whereas total protein concentration decreased in all the DEN treated groups as compared to the control groups. Biochemical parameters in case of combination groups were lower as compared to the DEN alone group. Grossly, group-2 animal showed hepatomegaly, liver paleness, cirrhosis and multiple nodules over the liver surface. However, in the combination groups the gross lesions of less intensity were observed. Microscopically, the liver of the rats of group-II revealed severe distortion of the hepatic cord arrangement, lobular pattern, bile duct proliferation, nuclear changes, fibrous tissue proliferation, inflammatory cell infiltration, Kupffer cell hyperplasia and degenerative changes. Similarly, in this group spleen showed coalescing lymphoid pulp, increase in the red pulp. Similar changes were noticed in the combination groups (group-3, 4, 5 and 6), but the changes were less severe in intensity as compared to the DEN alone group. Histopathologically, liver and spleen showed regenerative changes in plant treated groups. Immunohistochemical studies showed positive result for the presence of apoptotic bodies in DEN treatment groups. The results indicated hepatoprotective effect of plant extract in combination. DEN was found to be carcinogenic and produced significant hepatic nodules in rat model during in vivo experimentation.
Open Access
PATHOLOGY OF GASTROINTESTINAL AFFECTIONS IN BOVINE CALVES
(CSKHPKV, Palampur, 2016-07-25) Thakur, Vishal; Gupta, Vipin K.
Calfhood diseases have a major impact on the economic viability of cattle operations. The present study was conducted to investigate the pathology of gastrointestinal affections and to identify the major bacterial and parasitic etiologic factors responsible for gastrointestinal affections in calvesbyevaluation of necropsy and clinical samples. Out of 40 dead calves received for necropsy, 23 calves showed mild to severe enteritis and two cases hadconcurrent parasitic infestation due to Moniezia sp. Out of 23 cases of enteritis, 2 (8.70%), 17 (73.91%) and 4 (17.39%) cases showed enteritis, pnuemoenteritis and enteritis associated with other diseases, respectively. The histopathological changes in small intestine were infiltration of lymphocytes, macrophages, eosinophils in lamina propria and hyperemia of vessels. MNC infiltration was the prominent feature in all enteritis cases. In four cases submucosal edema was present. The villi exhibited variable changes including distortion, fusion and thickening. Lymph nodes showed mild lymphoid depletion in lymphoid follicles. Analysis of faecal samples revealed that out of 80 isolates in clinical and necropsy cases, E.coli, Citrobacter andSalmonella present in 80%, 18.75% and 1.25%, respectively. Isolation from 25 necropsycases showed presence of E.coli. A total of 20 E. coli isolates from necropsy cases were confirmed and serotyped for ‘O’ antigen from Central Research Institute, Kasauli. In diarrhoeic calves the prevalence of serotypes O89 and O120 were highest. A total of four serotype of O89 and three of O120 followed by two serotype of O9 and one each serotype of O119, O63, O88, O15, O118, O49 and O35 besides three untypeable and one rough. Disk diffusion test was used to check sensitivity of isolates from both clinical and necropsysamples. Cloxacillin was found most resistant drug while amoxy-sulbactum, ciprofloxacin and gentamicin were most effective drugs against the E.coli isolates. A total of 16 blood samples from diarrhoeic cases were analysed for total and differential leukocyte count, out of these four cases showed drastic increase in immature neutrophil count.Examination of 82 faecal samples shows three(3.65%)Strongyloidsspp, two(2.44%) Amphistomes sp. and four (4.82%) Eimeria spp.positive cases. PCR screening of the 23 faecal samples from necropsy cases for group A rotaviruswas found negative. In conclusion overall incidence of enteritis was 57.5%with highest numberin winters.
Open Access
PATHOLOGY OF RESPIRATORY DISEASES IN BOVINE CALVES
(CSKHPKV, Palampur, 2016-07-25) Pradhan, Stephanie S.; Gupta, V.K.
The present study was conducted to determine the prevalence and the factors responsible for respiratory diseases in bovine calves (sick-51, dead-40) of Himachal Pradesh. The nasal swabs and blood samples from sick and apparently healthy animals were collected from Palampur and its surrounding districts. The necropsy samples were collected from the postmortem received in the Department. Samples were collected from clinically sick and dead calves for bacteriological identification. Blood samples collected for TLC and DLC from calves exhibiting severe disease conditions revealed neutrophilia with increase in immature cells. Dead calves were subjected to necropsy examination and tissue samples were investigated at histopathology. Cytosmears prepared from nasal turbinate and affected portion of the lung revealed morphology of bacteria and the nature of exudate. In the present study, pneumonia was present in 31 (77.5%) of 40 calves and was considered an important contributor to death in 14(35%) of 40 calves whereas pneumonia associated with enteritis was observed in the remaining 17 (42.5%) calves. Out of 31 calves, 4 (12.9%) and 27 (87.1%) calves showed lesions suggestive of bronchopneumonia and interstitial pneumonia respectively. A few small white worms were observed in the upper respiratory tract in three cases, one with bronchopneumonia and two with interstitial pneumonia and the worms identified were Dictyocaulus viviparus. The most significant gross finding in the lung was hyperemia of the cranioventral region with pulmonary oedema. In 2 (6.45%) of 31 cases multifocal to coalescing abscesses were present in the cranioventral region. Mild rib impressions were also observed on the pleural surface of the lung in 13 (41.93%) cases with hazy appearance of the pleura in 24(77.4%) cases exhibiting both bronchopneumonia and interstitial pneumonia. The mediastinal and the bronchial lymph nodes showed gross changes of enlargement, oedema, hyperaemia and haemorrhage. On histopathological examination the turbinates showed mild inflammation in 27 (71.05%) out of 38 cases with infiltration of mononuclear (MNC) and polymorphonuclear (PMN) cells. Interstitial pneumonia was consistently characterized by the thickening of interalveolar septa due to MNC infiltration, hyperplasia of the bronchial and bronchiolar epithelium with peribronchial and peribronchiolar lymphoid proliferation. Clumping of macrophages was seen in nine cases with syncytia in one. In bronchopneumonia, the bronchioles and the alveolar lumen were filled with polymorphonuclear cells.Staphylococcusaureuswas isolated from the lungs showing subacute to chronic abscesses. Lymph nodes revealed mild to severe depletion of lymphocytes in the cortical region. E.coli, Streptococcus.sp, Staphylococcus.sp or Citrobacter.spwas common isolates from both upper as well as lower respiratory tract. Ciprofloxacin was found to be the most sensitive antibiotic followed by Gentamicin and Amoxycillin/Sulbactum. The present study revealed that pneumonia and pneumonia associated with enteritis are the common causes of calf mortality with its peak occurrence at six months of age.
Open Access
STUDIES ON THE EFFECTS OF PLANT BASED EXTRACT AGAINST Escherichia coli INFECTION IN BROILERS
(CSKHPKV, Palampur, 2016-07-25) Thakur, Shalini; Asrani, R.K.
The present study was planned with a view to observe the effects of methanolic extract of aerial parts of Artemisia nilagirica on growth, mortality, clinical signs, biochemical, pathological and ultrastructural changes in broiler chicks infected with E. coli. Three hundred thirty day old broiler chicks were randomly divided into six groups. The LD50 dose of E. coli was determined in a pilot experiment which was found to be 1x107 cfu ml-1. Plain E. coli infection, E. coli infection and methanolic extract (0.5g), E. coli infection and methanolic extract (1g), E. coli infection and methanolic extract (2g), methanolic extract (2g) and only feed was given to group(s) 1, 2, 3, 4, 5 and 6, respectively. E. coli infection was given on 7th day. Methanolic extract was given in water from day 4 to day 21. Results of present study showed that E. coli infected groups exhibited clinical signs of whitish diarrhea, anorexia, drooped wings, ruffled feathers, laboured and shallow breathing, huddling, dullness and depression. Higher values of ALT, AST, enzymatic creatinine, cholesterol, lower total protein and lower albumin values was observed in the plain infected group as compared to the control birds. However, in the combination groups (group 2, 3 and 4) the values were significantly less as compared the plain infected group (group 1). Grossly, in the group 1 fibrinous perihepatitis, fibrinous pericarditis, airsacculitis, splenomegaly and peritonitis were observed mainly. However, in the combination groups the gross lesions of significantly less intensity were observed. Microscopically, the liver and heart of group 1 revealed perihepatitis, pericarditis, vacuolar changes, leukocytic infiltration, necrosis and increased cytoplasmic granularity. Similarly in this group spleen showed reticuloendothelial hyperplasia, coagulative necrosis and lymphoid depletion. But the changes in the liver, heart and spleen were less severe in the combination groups. Significantly lower values were observed in group(s) 3 and 4. These changes suggest that Artemisia nilagirica has antimicrobial and hepatoprotective effects. Ultrastructural changes in the group 5 revealed relatively dense population of mitochondria along with abundant endoplasmic reticulum proliferation with no significant nuclear changes, as compared to control (group 6). These changes indicate that methanolic extract of Artemisia nilagirica is not toxic to liver even at the concentration of 2g per litre of water over a period of 18 days.
Open Access
STUDIES ON THE EFFECT OF CANNABIS SATIVA AGAINST DATURA STRAMONIUM- INDUCED TOXICITY IN RATS
(CSKHPKV, Palampur, 2016-12-14) Thakur, Nitika; Patil, Rajendra D.
The present investigation was undertaken to determine the efficacy of Cannabis sativa against Datura stramonium-induced toxicity in rats. A total of 72 (36 male and 36 female) weaned Wistar rats were randomly divided into six groups viz. Group-I served as normal control and received vehicle (NSS) only; Group-II rats treated with D. stramonium leaf extract @ 2000 mg/kg bw; Group-III rats treated with C. sativa root extract @ 1200 mg/kg bw; Group-IV rats treated with D. stramonium leaf extract @ 2000 mg/kg bw plus C. sativa root extract @ 400mg/kg bw; Group-V rats treated with D. stramonium leaf extract @ 2000 mg/kg bw plus C. sativa root extract @ 800 mg/kg bw; and Group-VI rats treated with D. stramonium leaf extract @ 2000 mg/kg bw plus C. sativa root extract @ 1200 mg/kg bw for 28 days by oral route. D. stramonium produced significant toxicity in rats from the first week of experimentation as seen by the clinical signs including typical nervous signs, mortality among 2 males and the significant reduction in the body weights on 14th and 21st day onwards in male and female rats, respectively. The absolute weights of kidneys, spleen and gonads were significantly decreased in female rats; however, relative organ weights of liver, kidneys, heart, brain and gonads were significantly increased in male rats. Haematological changes included significantly decreased PCV and MCHC and an increased MCV values seen in male rats. The serum activities such as alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, lactate dehydrogenase, serum enzymatic creatinine were significantly increased while, the decreased total protein levels and increased globulin and A:G ratio were noted in male and female rats. Gross pathological changes were less marked in group-II. However, histopathological changes were severe and consistently seen in Group-II rats with toxico-pathological damage in all the internal organs such as liver, kidneys, heart, brain, spleen, small intestine, mesenteric lymph nodes and gonads. No toxico-pathological changes were seen in the male and female rats given Cannabis sativa (Group-III) throughout the study. However, simultaneous administration of Cannabis to male and female rats from Groups-IV, V and VI was effective in alleviating the D. stramonium-induced toxicity in a dose-dependent manner.
Open Access
STUDIES ON IN VIVO EFFECT OF PLANT BASED EXTRACT AGAINST EXPERIMENTAL SALMONELLOSIS
(CSKHPKV, Palampur, 2017-06-29) Gautam, Hemlata; Asrani, R.K.
The present study was planned with a view to observe the effects of methanolic extract of roots of the plant Saussurea lappa on growth, mortality, clinical signs, biochemical and pathological changes in mice infected with a local isolate of Salmonella enterica serovar Typhimurium 4,12:i:1,2.The LD50 dose of S. Typhimurium was determined in a pilot experiment which was found to be 1.35x103 cfu ml-1.Two hundred mice were randomly divided into 7 groups, Group 1:plain control, Group 2: plant control with methanolic extract of Saussurea lappa @2000mg, Group 3: S. Typhimurium infection alone, Group 4: S. Typhimurium infection with Ciprofloxacin @200mg/kg, Group 5: S. Typhimurium infection with plant extract @200mg, Group 6: S. Typhimurium infection and plant extract @1000mg, Group7: S. Typhimurium infection and plant extract @2000mg.The mice in group 1, 2, 3 and 4 served as control. The methanolic extract of Saussurea lappa was given in drinking water to mice, 4 days before the infection and continued up to 10th of experiment. Results of the present study showed that mice of S. Typhimurium infected group exhibited clinical signs of ruffled hair coat, laboured breathing, staggering gait, hunched position, soiled anal area, soft faeces, reluctant to move and discharge from the eyes. ALT activity was found to increase, whereas total protein and albumin concentration decreased in all the infected groups as compared to the control groups. Values of creatinine was non-significantly different except in Ciprofloxacin group at 7DPI. Biochemical parameters in case of the combination groups were lower as compared to the plain infection group. Grossly group 3 animal showed hepatomegaly, hepatic necrosis, liver paleness and splenomegaly. However, in the combination groups the gross lesions of less intensity were observed. Microscopically, the liver of the mice of group 3 revealed severe necrotic foci, inflammatory cell infiltration, dilatation of sinusoids, vacuolar changes and increased cytoplasmic granularity. Similarly, in this group spleen showed congestion, necrotic foci, lymphoid depletion, granulocyte infiltration and RE cell hyperplasia. At 3 and 7 DPI, maximum lesions intensity scores were obtained. Similar changes were noticed in the combination groups (group 5, 6 and 7), but the changes were less severe in intensity as compared to the plain S. Typhimurium infected group. Histopathologically liver and spleen showed preservation of hepatocytes and hepatic cord structure and white pulp hyperplasia in plant treated groups indicated its antimicrobial, hepatoprotective and immunomodulatory effect. S. Typhimurium found to be highly pathogenic and produced significant hepatic lesion in mice model during in vivo experimentation.

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