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Open Access
STUDIES ON PATHOLOGY OF ENDOSULFAN TOXICITY IN BROILERS
(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2006-02) SUNEETHA, D; SRILATHA, Ch(MAJOR); SUJATHA, K; CHANDRA SEKHARA RAO, T.S
Open Access
PATHOLOGICAL CHANGES IN ARSENIC INDUCED TOXICITY AND ITS AMELIORATION IN BROILER CHICKEN
(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2009-12) TANJU, SAGI; MADHURI, D(MAJOR); ANJANEYULU, Y; DHANA LAKSHMI, K
ABSTRACT : The objective of the present work was to evaluate the toxic effects of sodium arsenite, As(III) and its amelioration with Moringa leaf powder @ 0.1% and ascorbic acid @ 200 ppm in broiler chicken. In the present study, chicks of day-old age belonging to Vencobb strain were assigned to six groups (I,II,III,IV,V,VI) consisting of eighteen chicks in each group. Group I served as control (basal diet) and birds in group II were given sodium arsenite @ 200 ppm in basal diet for 42 days. Group III and IV were given Moringa oliefera leaf powder and ascorbic acid @ 0.1% and 200 ppm in basal diet respectively. Group V was fed, diet containing sodium arsenite @ 200 ppm and Moringa leaf powder @ 0.1% for 42 days. Birds in group VI were given with sodium arsenite @ 200 ppm and ascorbic acid @ 200 ppm in basal diet for 6 weeks. Clinically, reduced feed intake and decreased body weight gain were observed in birds fed with sodium arsenite and improvement in these were noted in ameliorative groups. Six birds from each group were sacrificed at fortnightly intervals. Blood, serum and tissue samples were collected. Over all mean values of Hb,PCV and TEC levels in control groups were in normal range where as levels were significantly (P<0.05) decreased in toxin group. The serum biochemical assays showed significant (P<0.05) reduction in total protein, albumin, globulin, A/G ratio while significant (P<0.05) increase in AST, creatinine, GGT was observed in group II birds. Oxidative stress parameters revealed significant (P<0.05) decrease of liver GSH in group II. The ameliorative groups V and VI showed marked improvement in all the above parameters as compared to toxic group. Grossly group II birds revealed enlarged liver with hyperemia, haemorrhages on kidney and liver, reduction in the size of spleen and bursa of Fabricius. Histopathological lesions in group II comprised of severe sinusoidal and venous congestion with mononuclear cell infiltration. Kidney sections showed severe congestion, intertubular haemorrhages, hydropic degeneration, cystic spaces and necrosis of epithelium. Severe depletion of germinal centres was observed in sections of spleen. Depletion of lymphocytes and cystic spaces was observed in sections of bursa of Fabricius. Heart sections revealed complete disruption of muscle fibres and heavy mononuclear cell infiltration. Similar types of gross and histopathological lesions were noted in ameliorative groups of less intensity. In conclusion, sodium arsenite caused a significant toxicity in broiler chicken and its amelioration with Moringa oleifera leaf powder and ascorbic acid was effective in combating the arsenic induced toxicity.
Open Access
TOXICO PATHOLOGICAL STUDIES OF RAW KARANJ (Pongamia glabra vent) CAKE AND ITS AMELIORATION IN BROILER CHICKS
(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2009) ASHOK KUMAR REDDY, A; ANAND KUMAR, A(MAJOR)
ABSTRACT: Pongamia glabra vent, popularly known as karanj, belongs to Leguminaceae family which is a medium sized glabrous tree capable of growing under wide range of agro-climatic conditions. Karanj seed cake is the residue left after oil extraction, having scope for being used as protein source by replacing costly and scares soyabean meal or ground nut cake in poultry ration as it is rich in crude protein and is economical too. It’s incorporation in poultry ration is limited due to the toxic principle karanjin. The present study was undertaken to asses the effects of raw expeller pressed karanj seed cake (EKC) @10% of diet and efficacy of ameliorating agents methionine and activated charcoal @ 0.2% and 0.1% respectively. Day-old broiler chicks were divided into five groups consisting of 18 chicks in each group, various combinations of EKC, methionine and activated charcoal were incorporated as group I: control feed (CF), group II: CF + EKC @ 10%, group III: CF + EKC @ 10% + methionine @ 0.2%, group IV: CF + EKC @ 10% + activated charcoal @ 0.1% and group V: CF + EKC @ 10% + methionine @ 0.2% + activated charcoal @ 0.1% for 6 weeks of experimental period. All the experimental diets were isonitrogenous and isocaloric and they were fed through out the experimental period of 6 weeks and the influence of experimental diets on broilers was observed for performance, changes in haematological, serobiochemical parameters, gross and histopathological changes. Body weight gains and feed intake were recorded in each group at weekly intervals. Six birds from each group were sacrificed randomly at fortnight intervals. Blood and serum samples were collected prior to sacrifice for haematological and sero biochemical estimations and tissue samples were collected at necropsy for histopathological studies. Very few birds exhibited loose droppings irrespective of EKC or amelioration groups and mortality was not recorded in any of the groups. Significant (P< 0.05) reduction in the mean body weight gains, feed consumption and increase in FCR in EKC fed group was observed in comparison to control group. Improved body weight gains, feed consumption and feed efficiency in amelioration groups from 2nd week onwards till the end of the experiment in comparison to EKC fed group was observed but the results were still better in methionine amelioration group. Haematological findings revealed that overall mean values of Hb, PCV, TEC and TLC were significantly (P< 0.05) reduced in EKC fed group in comparison to control group, this might be attributed to disturbances in metabolism that lead to deficiency of clotting factors. The serobiochemical studies showed a significant (P< 0.05) reduction in glucose, total protein, albumin and globulin levels while significant (P< 0.05) increase in total cholesterol, creatinine and AST levels in EKC fed group in comparison to control group. The gross pathological findings in EKC fed group included enlargement of heart, liver, kidney and regression of spleen, bursa of Fabricius and congestion of liver and kidney at the end of 6th week. In amelioration groups lesions were mild at the end of 4th week but they almost normal at the end of the experiment. The histopathological changes of EKC fed group revealed severe disruption, degeneration and separation of myocardial fibers in heart, severe sinusoidal congestion, degeneration, necrosis of hepatic cells and diffused lymphoid aggregations in hepatic lobes. Kidney sections showed severe degeneration of tubules and glomeruli, diffused inter tubular haemorrhages and presence of hyaline casts, spleen showed severe lymphoid depletion, thickened trabecular arteries and presence of haemosiderin pigment and bursa revealed depletion of lymphocytes, separation & rupture of follicles and presence of big cystic spaces in the follicles at the end of 6th week. The amelioration groups showed mild lesions in various organs in comparison to EKC fed group which might be due to their protective effect against toxicity induced by EKC. Depending on the results of the present study can be concluded that methionine @ 0.2% is effective in comparison to activated charcoal @ 0.1% in ameliorating the toxicity of raw EKC.
Open Access
A PATHOLOGICAL STUDY ON GRADED LEVELS OF JATROPHA (Jatropha curcas) DEOILED SEED CAKE TOXICITY IN BROILER CHICKS
(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2009-01) ANIL KUMAR; ANAND KUMAR, A(MAJOR); ANJANEYULU, Y; RAJASHEKAR REDDY, A
ABSTRACT : Jatropha (Jatropha curcas) the medicinal plant of India has potential to solve the growing feed insecurity of world economically at cheaper rate. Jatropha seed cake is rich in crude protein, but proved to be toxic to poultry and livestock unless and until complete oil is not removed. But, the degree of toxicity varies at different concentrations. The present study was undertaken to study the pathological changes in broiler chicks fed with differently graded Jatropha deoiled seed cake. A total 120 day-old male broiler Vencobb strain chicks were divided into 5 groups consisting of 24 birds in each group. Group I was under control feed for the whole experimental period i.e., 6 weeks. Group II was fed with control fed (CF) for 1 – 14 days and then CF + JSC @5% for 15-42 days. Group III was fed with CF JSC @5% for 1 – 42 days. Group IV was fed with CF for 1 – 14 days and CF + JSC @10% for 15 – 42 days. Group V was fed with CF + JSC @10% for 1 – 42 days. The influence of treatment diets on broiler was evaluated in terms of performance, haematological, biochemical, gross and histopathological changes. Body weight gain and feed intake was recorded at weekly intervals in each group. Six birds from each group were sacrificed at fortnightly intervals. Blood, serum and tissue samples were collected for haematological, biochemical profile and hitopathological studies. Treatment groups resulted in significant (P<0.05) reduction in body weight gains, feed consumption and increase in FCR. The percentage mortality in groups III, IV and V was 12.5, 29 and 33 per cent, respectively and in groups I and II no mortality was observed. Clinical signs such as dullness, inappetance, loss of condition, greenish diarrhoea, reduced water intake and muscular and nervous imbalance prior to death in birds of group V were observed. Haematological study revealed that overall mean values of PCV, Hb, TEC and TLC were significantly (P<0.05) reduced in all the treatment groups. Group II and IV showed less reduction as compared to group III and V, respectively. The biochemical profile showed a significant (P<0.05) reduction in total protein, albumin, globulin, HDL cholesterol while significant increase in A/G ratio, ALT, AST, creatinine, bilirubin, total cholesterol and triglycerides was observed in all the treatment groups but the changes were less in group II and IV as compared to group III and V respectively. The gross pathological changes in treatment groups included hydropericardium, enlargement with rounded borders of liver, enteritis, regression in size of spleen and bursa of faricius. The histopathological changes included congestion and haemorrhages in various organs of treatment groups. Fatty changes were found in liver of group III, IV and V and in heart fatty changes were noted in group V. Heart sections revealed separated, disrupted muscle fibres. Liver sections revealed degenerative changes, necrosis in groups IV and V. Enterisis, infilitration of mononuclear cells in villi, broad and eroded villi observed in intestine. Focal shrinkage of glomeruli, degenerative ruptured tubules were noted in kidney sections. Sections of spleen revealed congestion and depletion of germinal centres. Bursa of fabricisus sections showed severe congestion and depletion of lymphoid follicles. The lesions described were more evident in group III and V as compared to group II and IV respectively as prolonged duration of toxicity in group III and V. The present study concluded that the toxicity of Jatropha at 10% level was severe in comparison to 5% level. At same level it’s toxicity was considerably less if the birds were maintained on normal feed for the 1st two weeks as the immune status of the birds will be sufficiently functional to withstand the toxicity.
Open Access
PATHOMORPHOLOGICAL STUDIES ON SHEEP LUNGS
(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2009-01) SANDHYA, M; MADHURI, D(MAJOR); ANJANEYULU, Y; GOPAL REDDY, A
ABSTRACT: Multiple etiological and environmental factors contribute to high susceptibility of sheep to pneumonia and pose trouble in control of this malady to sheep farming community. In present investigation, 1713 lungs were collected from slaughterhouses in and around Hyderabad. On detailed gross pathological examination, pneumonia was encountered in 248 lungs. The histopathological examination of lungs revealed interstitial pneumonia (29.03.%), fibrinous pneumonia (27.41%), broncho-pneumonia (22.58%), granulomatous pneumonia (9.67%), suppurative pneumonia (5.24%), Maedi (4.83%), aspiration pneumonia (0.80%), Jaagsiekte (0.40%) and other common conditions like congestion, edema, haemorrhages, emphysema, bronchiectasis etc in (2.01%) cases respectively. In interstitial pneumonia lungs appeared voluminous and enlarged. Histologically, marked thickening of the interalveolar septa and interlobular septa due to haemorrhages and infiltrations of polymorphonuclear cells and mononuclear cells observed. In fibrinous pneumonia lungs were hyperemic and haemorrhagic, stringy, copious exudate was noticed in the bronchioles. Microscopically, fibrinous exudate mixed with neutrophils and macrophages were seen in the alveolar lumen. Bronchopneumonia was characterized by pulmonary tissue showing patchy to diffuse areas of consolidation ranging from red to grayish colour prominent on cranial lobes and copious bronchial exudation. Microscopically, lining epithelium of the bronchioles were hyperplastic with varying amount of exudation in the lumen. Marked mononuclear cells in bronchiolar sub-mucosa and in adjoining alveoli was noted. Granulomatous pneumonia caused by Mycobacterium was noticed as tuberculosis. Localized lesions with caseation were seen in upper lobes of lungs. Histological examination revealed characteristic granulomatous inflammatory reaction forming both caseating and non caseating tubercle. In suppurative pneumonia lungs showed purulent nodules and microscopically heavy neutrophils and cellular debris was observed in the lumen of alveoli. In Maedi, lesions were of interstitial pneumonia type. The lungs were pale and consolidations were prominent. Microscopically, large lymphoid cell aggregates around most of the bronchi, bronchioles and blood vessels was the prominent lesion noticed in all the Maedi affected lung sections. In aspiration pneumonia lungs showed congestion and consolidation. Microscopically the pleura were thickened, edematous and congested. The parenchymatous tissue around the foreign debris was necrosed. Jaagsiekte was characterized by lung consolidation with a few grayish white nodular foci. Histologically normal thin alveolar cells were replaced by cuboidal or columnar cells with papillary projections into the alveoli. Microbial isolation was attempted from some pneumonic cases. The major pathogenic bacteria isolated from bronchopneumonia were E. coli, Pasteurella multocida and Staphylococcus aureus. In fibrinous pneumonic cases pathogenic bacteria isolated were Staphylococcus, E.coli, Pasteurella multocida and Mycobacterium, E.coli, Staphylococcus and Pasteurella multocida were isolated in 16 of 107 cases of interstitial pneumonia. Pathogenic bacteria Mycobacterium tuberculosis was isolated in cases of granulomatous pneumonia. The results of the present study suggested pneumonia could be one of the important cause of death in sheep in Hyderabad. Secondly incidence of granulomatous pneumonia due to mycobacterium spp. has been recorded in 16 lungs (28.57%) suggesting prevalence of TB in sheep in Hyderabad. As this disease causes huge economic losses due to decline in lifetime productivity and death of animal in advance stage of disease. Therefore further studies are to be done to study the seroprevalence of tuberculosis infection in sheep.
Open Access
PATHOLOGICAL STUDIES ON ENDOSULFAN TOXICITY IN RATS AND ITS AMELIORATION WITH OCIMUM SANCTUM
(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2008-11) BHARATH, B.K; ANJANEYULU, Y(MAJOR); Srilatha, Ch
ABSTRACT : India has highest animal population in the world and animal husbandry occupies a prominent position in agricultural sector.It has been one of the subsidiary occupation for rural people offering socioeconomic stability and is one of the major contributors to the national economy. Indiscriminate use of pesticides to control pests in various agricultural and animal husbandry practices and vector control programmes of public health concern may lead to health hazards due to their presence in food of human beings and livestock. Endosulfan is a chlorinated hydrocarbon pesticide under the cyclodiene subgroup. It emerged as a leading chemical used against a broad spectrum of insects and mites in agriculture and allied sector. It acts as contact and stomach poison and has slight fumigant action. It is used in vegetables, fruits, paddy, cotton, cashew, tea, coffee, tobacco, timber crops and also as a wood preservative. Exposure to endosulfan can induce a number of effects including neuro, hepato and nephrotoxicity, haematological and biochemical effects, altering the immune system and damage the reproductive organs. Most of the earlier studies are restricted to acute effects of pesticide in experimental animals. However the situation in practice is not toxicity resulting from a single or a few large doses of a given pesticide, but due to oral intake of very small quantities over a reasonably long period of time. Studies related to endosulfan toxicity have been carried out in different animals. There is paucity of detailed information regarding endosulfan induced chronic toxicity and also ameliorative effect of O S against endosulfan, hence a systematic work was undertaken to study the toxic effects of low levels of endosulfan and ameliorative effect of Ocimum sanctum against endosulfan induced damages and immuno suppression with the objectives to study the effect of endosulfan on haematobiochemical, gross and histopathological changes in different organs and to study the ultrastructural changes in liver and RBCs and ameliorative effect of Ocimum sanctum against endosulfan induced damages. The present experiment was designed to make a systematic study of experimentally induced endosulfan toxicity in male rats. Technical grade endosulfan was given at 6, 3 and 1.5 mg / Kg b.wt to groups II, III and IV by mixing in ground nut oil for 6 weeks, to the groups V, VI and VII in addition to endosulfan, Ocimum sanctum was given @200 mg / Kg b.wt daily per orally for the same duration. Group I and VIII served as oil and Ocimum sanctum control. Specific clinical signs were not observed, except few rats of the Group II and Group V. Showed almost similar clinical signs in general during the experimental period which included dullness, depression, diarrhoea, intermittent anorexia and hyper excitability. In the present investigation no mortality was observed in any of the treated group. Significant (P<0.05) reduction in the weight gain was observed in toxin treated groups when compared to control groups and these values are nearer to normal in lower doses of endosulfan with OS ameliorated groups. Significant (P<0.05) reduction in the PCV, Hb, TEC, TLC, percent lymphocytes count and significant increase in the percent neutrophil count were observed in all the endosulfan treated groups. These counts were significantly improved in the OS ameliorated groups with lower dose of endosulfan (groups VI and VII). Significant (P<0.05) increase in serum glucose, creatinine, cholesterol, ALT, AST, and significant (P<0.05) decrease in the TSP levels in endosulfan treated groups when compared to control groups in a dose dependant manner. And these levels were in between the above groups in the OS ameliorated groups. Significant (P<0.05) decrease in the serum T3 and T4 levels were noticed in all toxin treated groups when compared to the control groups in a dose dependent manner, and there was no change in TSH levels in any of the groups. In OS treated groups significant improvement was observed compared to the toxin treated groups in T3 and T4 levels. Significantly (P<0.05) decreased testicular wet weights, sperm counts were observed in all the experimental group when compare to the Oil control group. Significant decrease in the HA titer (log) and DNCB contact sensitivity score in groups II, III and V when compare to the control groups and these values are in between the above groups in IV, VI and VII groups. In the present study gross lesions were prominently observed in the liver followed by kidney, lung, spleen, brain and heart. Liver revealed moderate enlargement during the 2nd week. Severe enlargement of the liver with rounded borders and paleness of the liver was observed after 4th and 6th week in all endosulfan treated groups in a dose dependant manner and changes are less severe in OS ameliorated groups. Kidneys revealed enlargement with moderate congestion in all the endosulfan treated groups throughout the experimental period. Lungs of the rats from the endosulfan treated groups revealed moderate to severe congestion throughout the experimental period. With less severity in OS treated groups. Severe enlargement of spleen was observed during the 2nd week of the experiment but after 4th and 6th week observed the reduction in the size of the spleen in endosulfan treated groups with less severity in the OS groups. Brain showed mild to moderate congestion in all the experimental animals throughout the experiment. Mild enlargement of the heart was observed in all the endosulfan treated groups throughout the experimental period. Histopathologically the liver of rats fed with endosulfan for 2 weeks revealed congestion, degenerative changes in hepatic cells with cellular swelling, mild fatty changes, focal MNC aggregation, mild perivascular and periportal MNC infiltration, sinusoidal dilatation with haemorrhages and focal loss of hepatocytes with MNC infiltration and mild bile duct proliferation. In addition to above changes mild to moderate fatty changes, few areas of hyper chromatic nuclei, bi nucleated hepatocytes, loss of hepatocytes with dilated sinusoids and micro granulomatous lesions, MNC infiltration around the blood vessels with fibroblast proliferation by the end of 4th week were noticed. More number of bi nucleated hepatocytes, individualization of hepatocytes, bile duct proliferation with hyperplasia of epithelium, karyomegaly and focal areas of centri lobular necrosis were observed very conspicuously by the end of 6th week in a dose dependant manner and these lesions were mild in the OS ameliorated groups with lower doses of endosulfan (group VI and VII). Microscopic examination of kidneys treated with endosulfan revealed dose dependent congestion of blood vessels and glomeruli, haemorrhages in between the tubules, mild fibroblast proliferation, degenerative changes in the tubular epithelium and mild infiltration of MNCs by the end of 2nd week. In addition desquamation of epithelium leaving only basement membranes, severe haemorrhages, loss of tubules leaving cystic spaces, perivascular edema with fibroblast proliferation, karyomegaly, focal aggregation of MNCs and atrophy of glomerular tuft were observed by the end of 4th week. In addition hyaline casts in the lumen of tubules, some tubules with hyperplasia of tubular epithelium, in some areas total loss of tubules with haemorrhages and lytic changes around the glomeruli and capillaries and cystic glomeruli were observed by the end of 6th week. All these changes were dose and time dependent and these changes are less severe in OS treated groups in lower dose groups. Congestion, thickened blood vessels, mild to moderate edema, perivascular and peribronchiolar edema, haemorrhages, emphysematous changes and inflammatory cell infiltration were observed by the end of 2nd week. In addition to above hyperplasia of bronchiolar epithelium, hyalinised blood vessels, peribronchiolar fibrous tissue proliferation, thickened interstitial space due to infiltration of MNCs, RBCs and fibroblast by the end of 4th week were noticed. Hyper trophy and hyperplasia of bronchiolar lymphoid follicles, MNCs infiltration around the blood vessels, peri bronchial regions aggregation of the macrophages, plasma cells and other MNCs and haemorrhages around the blood vessels were observed by the end of 6th week all these changes are dose and time dependent. In OS ameliorated groups these changes were mild compare to the corresponding toxin treated groups. Cerebrum of rats treated with 6mg/kg b.wt endosulfan showed congested blood vessels, focal minute haemorrhages in the cerebrum, by the end of 2nd week. In addition proliferation of capillaries, mild to moderate gliosis, shrinkage of neurons, central chromatolysis, neuronophagia, micro nodular lesions in the cerebrum, sub meningeal haemorrhages in cerebrum and cerebellum region, prominent chromatolysis, gliosis, neuronophagia, demyelinating changes, vacuolation, shrinkage of neurons, focal loss of grey matter with aggregation of glial cells by the end of 4th week. Sub meningeal accumulation of MNCs, sever gliosis, vacuolation, thickening of meninges with fibroblasts, in certain areas and Purkinjee cell hyperplasia were observed, Central chromatolysis of neurons was more conspicuous in rats sacrificed by the end of 6th week. Cerebellum of rats congested blood vessels, focal minute haemorrhages focal loss of purkinjee cells, sub meningeal haemorrhages, grouping of purkinjee cells and central chromatolysis at the end of 4th and 6th week. Testis showed congestion, interstitial edema, separation of seminiferous tubules and degenerative changes in the seminiferous tubules by the end of 2nd week. Perivascular edema, loss of tubular epithelium leaving only basement membranes with sertoli cells and degeneration with desquamated cell debris within the lumen of tubules, MNCs and plasma cell infiltration in the interstitial spaces, hyalinization of blood vessels and few seminiferous tubules were observed by the end of 4th week. Complete separation of germinal epithelium from basement membrane, severe degenerative and lytic changes were observed by the end of 6th week. All these changes were dose and time dependent and these changes were severe in OS treated groups. Spleen of group II and III rats revealed congestion, mild lymphoid depletion by the end of 2nd week, engorgement of red pulp with erythrocytes, severe congestion of vessels with moderate to severe depletion of lymphocytes and reduced number of lymphoid follicles at 4th week to 6th week was observed. In other groups mild lymphoid depletion with congestion of vessels was observed and in spleen of group VII rats only mild depletion was observed when compare to the control groups. In group II and V rats revealed changes like congestion of blood vessels, mild to moderate increase in number of goblet cells, edema, desquamation of epithelium and areas necrosis with MNCs infiltration by the end of 6th week. Group III and VI rats revealed changes like mild increase in number of goblet cells and mild mononuclear cell infiltration. These changes were mild in OS ameliorated groups compared to toxin treated groups. Pancreas of all the endosulfan treated rats showed dose dependant congestion of blood vessels, haemorrhages and mild degenerative changes both in acini and Islets of Langerhans by the end of 2nd week. In addition hyalinised blood vessels, infiltration of MNCs within and between the follicles, severe vacuolar degeneration of both endocrine and exocrine part, atrophy and necrosis of Islets, epithelial hyperplasia of intercalated ducts, fibroblast proliferation around the blood vessels, ducts and in between the follicles, atrophy of the acini were observed by the end of 6th week. Similar lesions were observed in the group V as that of group II but these changes was less severe in group VI and VII. Adrenals cortex of endosulfan treated groups showed dose dependent vacuolar degeneration, telengectasis, swollen foamy cytoplasm, few bi nucleated cells, sub capsular and medullary haemorrhages. Very mild changes were observed in OS ameliorated groups. Lymph nodes of all the experimental groups showed dose and time dependent congestion, haemorrhages, edema and cortical depletion. And the depletion was not so severe compare to the endosulfan treated alone in the lower dose with O S ameliorated groups. Scanning electron microscopy of RBCs treated with endosulfan revealed dose dependent changes like loss of normal cellular outlines with membrane fusions, crenations, surface pits, loss of biconcavity and threading of membranes by the end of 4th and 6th week. Where as in OS treated groups these changes were restored towards normal. Transmission electron microscopy liver treated with endosulfan revealed swollen nucleus, clumping of outer and inner nuclear membrane, scanty chromatin material, vacuolation of cytoplasm, reduction in the number of mitochondria and endoplasmic reticulum, fragmentation and condensation of chromatin material by the end of 4th and 6th week on dose dependant manner compare to the control. In higher dose levels of toxin in the amelioration dense presence of mitochondria and endoplasmic reticulum were noticed.
Open Access
STUDIES ON PATHOLOGY OF INDUCED FENVALERATE TOXICITY IN RATS WITH SPECIAL REFERENCE TO NEUROENDOCRINE SYSTEM AND ITS AMELIORATION WITH WITHANIA SOMNIFERA (ASHWAGANDHA)
(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2008-11) AMARAVATHI, P; SRILATHA, Ch(MAJOR); RAMADEVI, V; SURESH KUMAR, R.V
ABSTRACT : The effects of pesticides have been recognized as a serious public health concern during the past decades. Pyrethroids and organophosphates have a wide spectrum of insecticidal potency and produce acute toxicosis in vertebrates manifested by the inhibition of acetylcholine esterase (Mohamed et al. 1993). Fenvalerate is one of this groups which is potent insecticide that has been in use since 1976. Over the last 20 years, large amounts of formulated fenvalerate, approximately 1000 metric tones per year have been used as an agricultural pesticide (WHO, 1992). Although fenvalerate considered having low acute toxicity to mammals, it is having severe neurotoxic effects and estrogenic activity causing endocrine disorders. Hence fenvalerate is enlisted as one of the neuroendocrine disrupting chemicals (EDCs) and has aroused world wide concern. EDCs are known to act at multiple sites through multiple modes of action, but the mechanism of action is poorly understood (Junhe et al. 2006). Persual of literature revealed limited information regarding toxicopathological effects of fenvalerate with special reference to neuro endocrine system (Mani et al. 2002). Organochlorines and organophosphates have been proved to protect oxidative stress and very few reports are available regarding involvement of reactive oxygen species (ROS) in pyrethroid toxicity. Beneficial influence of vitamin E and vitamin C are well documented in reducing the harmful effects of fenvalerate and limited information is available regarding the herbal products in ameliorating the pyrethroid toxicity. Keeping in view of its wide spread use, the present study is taken up to investigate toxicopathological manifestations. The present study was carried out by procuring 144 female rats and were randomly divided into eight groups consisting of 18 rats in each group. Fenvalerate (20% EC) gavaged per orally using ground nut oil as vehicle @ 42.5, 21.25 and 10.125 mg / kg b.wt. to groups II, III and IV respectively and ashwagandha @ 200 mg / kg b.wt. in distilled water was given along with fenvalerate to group V, VI and VII for 6 weeks to study ameliorative effects. Group I and VIII kept as control for toxin and ashwagandha fed groups respectively. Six rats from each group were sacrificed at fortnight interval. Dullness, depression, pawing, burrowing, hyper excitability, piloerection, salivation, lacrimation, clonic seizures and whole body tremors were the main clinical signs in all toxin fed and ameliorated groups. Significant (P<0.05) decrease in the body weight gains was observed in higher dose levels of fenvalerate fed groups (group II and III) when compared to control groups (I &VIII). In ameliorated groups no significant difference was observed when compared to corresponding toxin treated groups. Significant (P<0.05) decrease in TEC, PCV, haemoglobin and TLC values was observed in all fenvalerate treated groups when compared to control groups. There was no significant difference among treated groups and ameliorated groups. There was no significant difference in ALC and ANC values. Significant (P<0.05) decrease in serum total protein, serum cholesterol and serum acetyl choline esterase and significant increase in serum AST, ALT, serum glucose and creatinine levels were observed in all fenvalerate treated groups There was no significant difference between toxin treated groups and ameliorated groups. But in group VII significant improvement was observed when compared to corresponding toxin treated group. There was significant (P<0.05) decrease in T3, T4 and E2 levels in all fenvalerate treated groups when compared to control groups and group VII. In between groups significant variation was observed. Significant improvement was observed only in lower dose levels of toxin with ameliorating agent (group VII). Significant decrease in HA titers and DNCB dermal sensitivity was observed in all toxin fed groups and indicating decrease in cell mediated immunity and humoral immunity. Grossly mild changes were noticed only in liver, lung, spleen and Brain. Histopathologically lesions were prominently noticed in kidney followed by liver, lung, brain, ovary, uterus, heart, spleen, lymph node and intestine in all toxin fed groups. In kidneys congestion, edema, degenerative changes, intertubular haemorrhages, mild fatty changes in tubular epithelial cells were observed. Glomerular atrophy and fibrous tissue proliferation around glomeruli were consistently recorded in fenvalerate treated groups in a dose dependent manner. Cystic tubules and glomeruli were observed in majority of higher doses of fenvalerate treated rats by the end of experiment. In ameliorated groups also similar changes were observed except in group VII where mild congestion and mild degenerative changes were observed. Liver revealed congestion, degenerative changes, haemorrhages, mild fatty changes, infiltration of MNCs, loss of hepatocytes and proliferation of bile ducts in all fenvalerate treated groups.. Apart from prominence of lesions in group II, other changes observed were individualization of hepatocytes, perivascular infiltration of MNCs and hyper chromatic nuclei in some hepatocytes. In ameliorated groups also similar changes were observed except in group VII where mild sinusoidal haemorrhages and mild degenerative changes were observed. In lungs moderate to severe congestion, edema, perivascular and peribronchiolar infiltration of mononuclear cells and hypertrophy of lymphoid follicle was observed. In many rats areas of emphysema, thickened alveolar septa due to edema, infiltration of MNCs, RBCs, macrophages and few plasma cells, hyalinised blood vessels and giant cells in alveolar lumen was observed prominently in higher doses of fenvalerate treated groups (group II and III). In lower doses of toxin (group IV) only mild pneumonic changes were observed. In ameliorated groups also similar changes were observed except in group VII where mild congestion with focal infiltration of MNCs was observed. Histopathologically cerebrum revealed congestion, submeningeal haemorrhages and demyelinating changes in all fenvalerate treated groups. In addition gliosis, central chromatolysis and proliferation of capillaries were observed in cerebrum of group II and III rats. Sub meningeal haemorrhages, focal loss, shrinkage, central chromatolysis of purkinjee cells in the purkinjee cell layer and mild congestion and haemorrhages in granular layer of cerebellum were observed more conspicuously in group II and III rats. In ameliorated groups also similar changes were observed except in group VII where only mild changes were noticed. Microscopic examination 0f heart revealed mild to moderate haemorrhages and congested vessels and mononuclear cell infiltration in between cardiac muscle fibers. In addition to above lesions, sarcolysis along with eosinophilic cellular infiltration were observed conspicuously in higher doses of fenvalerate treated groups. Similar changes were observed in ameliorated groups (group II and III). Microscopic lesions in intestine were not characteristic in toxin treated rats except moderate increase in number of goblet cells, necrosis and hyperplasia of intestinal epithelial cells in group II and III. Histopathological examination of spleen revealed congestion, focal areas of hemorrhages, mild lymphocytolysis and engorgement of red pulp with erythrocytes in group II and III. In lymph node congestion of blood vessels and mild to moderate lymphocyte depletion was observed group II and III. In pancreas degenerative changes in islets of Langerhans, serous acini, necrosis, congestion, haemorrhages, infiltration of MNCs loss of acini with fibrous tissue proliferation was observed in group II and III. In adrenals sinusoidal and severe medullary haemorrhages and degenerative changes in medulla were observed in all fenvalerate treated groups. In ameliorated groups similar changes like those of corresponding toxin treated groups were observed in intestine, spleen, lymph node, pancreas and adrenals. In ovary severe haemorrhages, hyaline fibrosis, increased number of atretic follicles, degenerated follicles and decreased number of primordial follicles were observed in all toxin treated groups in dose dependent manner. Similar changes were observed in ameliorated groups. Uterus revealed congested blood vessels, haemorrhages, hyperplasia of endometrial epithelium, periglandular edema and eosinophilic cell infiltration with few plasma cells, lymphocytes in sub mucosa and atrophy of endometrial glands and periglandular fibrous tissue proliferation in all fenvalerate treated groups in dose dependent manner. In ameliorated groups similar changes were observed. Ultra structurally dose dependent marked distortions in membrane, irregular shape with many small shallow pits and small perturbations on the surface were noticed by the end of 6th week in all toxin treated groups when compared to control. In ameliorated groups mild distortion was observed. In liver swollen nucleus, margination of chromatin material, thickened nuclear membrane, vacuolation of nucleoplasm, vacuolation of cytoplasm, uniform reduction of size of mitochondria and mild fatty deposition were noticed in all toxin treated groups in dose dependent manner. In ameliorated groups mild to moderate changes were observed when compared corresponding toxin treated groups.
Open Access
PATHOLOGICAL STUDIES OF EXPERIMENTALLY INDUCED COMBINED (ISN + RIF) DRUGS TOXICITY IN RATS AND PROTECTIVE EFFECTS OF VITAMIN-E AND WITHANIA SOMNIFERA
(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2008-10) ARUNDHATHI, S; ANAND KUMAR, A(MAJOR); ANJ ANEYULU, Y; GOPALA REDDY, A
ABSTRACT: A total of 24 healthy male Wistar rats (age 60-100 days, weighing 175-260mg) procured for the present experimental study and the experiment was carried out according to the guidelines and prior approval of Institutional Animal Ethics Committee. Animals were divided into four groups consisting of 6 in each group. The experimental study was designed as follows:- Group I-Control, Group 11-ISN@SOmg/kg body wt + RIF@lOOmg/kg body wt once daily orally for 21 days, Group 111-ISN@SOm&g body wt + FUF@lOOrng/kg body wt once daily orally for 21 days + Withania sornnifira@lOOOmgkg feed daily for 21 days, Group N-ISN@SOm&g body wt + RIF@lOOmgkg body wt once daily orally for 21 days + vitamin E@300mg/kg feed daily for 21 days. Whole blood and serum was collected 4 times at weekly intervals (0, 7, 14 and 21 days) for estimation of various haematological and biochemical parameters and analyzed statistically. Haematological studies revealed that overall means of PCV, Hb and TEC were significantly (P<0.05) reduced and TLC was increased significantly (Pc0.05) in group I1 in comparison to other groups. The biochemical assays showed a significant (Pc0.05) reduction in total protein, HDL cholesterol, while significant (Pc0.05) increase in AST, ALT. GGT, total cholesterol and triglycerides in group 11. The - tissue enzyme assays revealed a significant (P<0.05) increase in TBARS and significant (P<0.05) decrease of GSH values in group 11. The ameliorative groups 111 and IV showed significant improvement in all parameters in comparison to group 11. The gross pathological changes in the group I1 animals included mild hepatomegaly. rounded borders, congestion and in heart petechiae and ecchymotic haemomhages with focal haemorrhages on kidney. The histopathological changes in group I1 revealed enlarged portal tracts with infiltration of lymphocytes, severe sinusoidal congestion and fatty change in perigortal areas. Hepatocytes revealed degeneration and moderate to severe necrosis. Heart revealed disruption and separation of muscle fibres, endocardid haemorrhages, moderate infiltration of lymphocytes and degeneration of muscle fibres, necrosis and fragmentation. In kidney glomeruli revealed reduction in glomerular space, vacuolation in few glomeruli and degeneration with extensive haemorrhages in interstitial tubular spaces, tubular epithelial cells showed fatty change with disrupted tubules and focal areas of tubular atrophy. Brain revealed congestion, haemorrhage and infiltration of mononuclear cells in meningeal vessels surrounding areas fatty change and increased cellularity in granular cell layer of cerebellum. Group 111 and IV animals revealed mild infiltration of mononuclear cells in and around portal areas with mild sinusoidal congestion. Structural details were intact and mild fatty changes were evident. Heart revealed mild range of degeneration and disruption of muscle bundles. Kidney showed mild haemorrhages in interstitial spaces and normal glomerular space. Brain revealed decreased cellularity and mild congestion in and around the meningeal areas. The present study concluded as follows: I. The changes resulted by the antitubercular drugs in the haernatology can be attributed to the hepatic damage. 2. The biochemical changes might be attributed to the toxic metabolites of the isoniazid and rifampicin. 3. The gross and the histological changes in the study might be due to the toxic reactive metabolites of the drugs which binds to cellular macromolecules and release or form toxic free radicals inturn caused the tissue damage. 4. Supplementation of Withania somnifera (1%) resulted in significant improvement in the haematological and biochemical parameters might be due to the antioxidant, antistress, liver tonic and anti-inflammatory properties. 5. Supplementation of vitamin E (0.3%) resulted in significant improvement in the haematological and biochemical parameters which can be attributed to the antioxidant and anticholesterimic effects. 6. Elevation of tissue TBARS and reduction of GSH in the study might be because of increased lipid peroxidation due to toxic reactive metabolites of the drugs. The present study indicated that Withania somnifera @ 1OOOmgkg feed and vitamin E @300mg/kg feed were effective in counteracting the toxic effects of the antitubercular drugs but not up to the required levels. Keeping this in view, Wer studies can be advocated using different dose rates and different routes of administration to overcome the affects of antitubercular drugs which would be beneficial for conservation of rare species on the earth.
Open Access
PATHOLOGY OF NEOPASMS IN DOGS
(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2008-10) CHANDRAVATHI, T; ANJANEYULU, Y(MAJOR); ANAND KUMAR, A; NARASIMHA REDDY, Y
ABSTRACT : Cancer is the most urgent problem of the contemporary medical field. In most of countries it is the second leading cause of the death, next to the cardiovascular disease. Neoplasm is a leading cause of death in all species. In animals dogs have highest incidence. About 45% dogs reaching middle age (6-7 years) will either develop a tumour, suffer medical complication as a result of tumour, or die as a result of neoplastic disease so there is need to emphasise on canine neoplasms. The present study was carried out to know age, breed and sex wise incidence of canine neoplasms in and around Hyderarabad and samples were collected from different hospitals after surgical incision. Immunohistochemistry was carried out to know the proliferating activity in different tumours. A total of 68 samples were collected, out of which 31 (45.59%) were benign and 37 (54.41%) were malignant tumours. Tumours were classified into epithelial 40 (58.83%), mesenchymal 20 (29.40%), round cell 5 (7.36%) and mixed tumours 3 (4.41%). The highest risk of development of various tumours was found in the age group of 7-9 years, followed by 4-6 years, above 9 years and below 3 years and the incidence was 29 (42.65%), 20 (29.41%), 19 (27.94%) and 1 (1.47%) respectively. The frequency of occurrence of neoplasms was slightly higher in females 38 (55.88%) compared to the males 30 (44.12%). Among the breeds affected Pomaranian breed represented more with 18 (26.50%) followed by non-descriptive 16 (23.52%), German Shepherd 16 (23.52%), Labrador 6 (8.82%), Doberman 6 (8.82%) Dachshund and Rottweiler 2 (2.94%), each one of Collie and Great Dane (1.47%). The organ wise incidence of tumours, included skin and mesenchymal tumours 29 (42.65%), mammary tumours 17 (25.0%), joint and bone tumours 9 (13.23%), vaginal tumours 5 (7.35%), testicular tumours 4 (5.88%), vulva and penis 3 (4.41%), ovarian tumours 2 (2.94%) and each one of transitional cell carcinoma and prostatic carcinoma (1.47%). Benign tumours include, fibroma 3 (4.42%), sebaceous gland adenoma 2 (2.94%), perianal gland adenoma 2 (2.94%), papilloma 2 (2.94%), melanoma 2 (2.94%), canine cutaneous histiocytoma 2 (2.94%), benign mammary tumours 2 (2.94%), hemangiopericytoma 2 (2.94%), lipoma 2 (2.94%), fibromatous epulis 2 (2.94%), mammary gland adenoma 1 (1.47%), apocrine gland adenoma 1 (1.47%), anal sac adenoma1 (1.47%) myxoma 1 (1.47%) and hemangioma 1 (1.47%), kaposi like tumour 1 (1.47%), leiomyoma 1 (1.47%), fibroleiomyoma 1 (1.47%), fibromyxoma 1 (1.47%), and fibrolipomyxoma 1 (1.47%). Malignant tumours included mammary gland carcinoma 14 (20.59%), squamous cell carcinoma 3 (4.42%), transmissible venereal tumour 3(4.41%), sertoli cell tumour 3 (4.41%), fibrosarcoma 2 (2.94%), osteosarcoma 2 (2.94%), ovarian adeno carcinoma 2 (2.94%), basal cell carcinoma 1 (1.47%), malignant melanoma 1 (1.47%), seminoma 1 (1.47%), prostatic carcinoma 1 (1.47%), transitional cell carcinoma 1 (1.47%), chondrosarcoma 1 (1.47%), rhabdomyosarcoma 1 (1.47%) and lymphangiosarcoma 1 (1.47%). Oxidative stress is the central mechanism in the pathogenesis of many diseases including cancer. The present study included the estimation of TBARS, SOD, catalase, GSH and GST. The TBARS, SOD, GSH, GST and catalase levels significantly (P<0.05) increased in the tumour tissue compared to the normal tissues. The number of AgNOR’s had been associated with cell proliferation. The mean AgNOR dots varied from 2.81±1.02 to 16.89±2.58 in individual tumour. The mean number of AgNOR in benign tumours was significantly (P>0.05) lower than the malignant tumours.. The lowest AgNOR count was observed in adenoma of mammary gland (2.81±1.02) and highest in cutaneous histiocytoma (16.89±2.58). In case of mammary tumours highest AgNOR counts was recorded in anaplastic carcinoma of mammary gland (10.58±2.67). PCNA was associated with the cell proliferation in different tumours. The mean PCNA positive nuclei in tumour tissues varied from 8.23±1.25 to 437.95±8.54. The lowest PCNA index was observed in myxoma while highest PCNA index was observed in cutaneous histiocytoma. The mean number of PCNA positive nuclei in benign tumours was significantly (P<0.05) lower than the malignant tumours. Anaplastic (336.95±6.98) and solid (221.38±4.78) mammary adenocarcinomas showed highest PCNA counts than cystic papillary (93.15±7.89) and tubular adenocarcinomas (77.42±5.74). There exist a significant (P<0.01) correlation between AgNOR counts and PCNA index.

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