H.S. Banga

Ameliorating Effects of Vitamin E and Selenium on Immunological Alterations Induced by Imidacloprid Chronic Toxicity in Chickens

The purpose of the current study was to investigate the immunological impacts of chronic imidacloprid insecticide toxicity in broiler chickens and the protective effects of vitamin E and selenium. Broiler chicks (n=90) aged day old were randomly segregated into three groups of 30 chicks each and were kept in separate pens. Newcastle disease (ND) vaccine was given on day 7 and 28 of age. On day 8, chicks in group I were administered 5 mg/kg bw (1/20 LD50) imidacloprid orally. Chicks in group II were given imidacloprid 5 mg/kg bw orally plus a mixture of vitamin E and selenium ≅ 200 mg/kg diet, while group III was given distilled water (DW) orally and served as a control. The treatments were given daily based on weekly body weight till day 45. Antibody titration, serum total immunoglobulins, circulating immune complexes, cell-mediated immunity and histopathology of bursa of Fabricius and spleen were examined. Imidacloprid produced significant decline in the titer of antibodies against ND vaccine, total immunoglobulins and circulating immune complexes in imidacloprid treated group on day 45 as compared to control group. There were no significant changes in the skin thickness between treated chickens and chickens of control group. Histopathology of the bursa of Fabricius revealed edema, lymphocytic depletion in the medulla and cortex and mild interfollicular fibrosis in imidacloprid treated group. The spleen showed mild haemorrhages and lymphocytic depletion. Supplementation of vitamin E and selenium resulted in marked improvements in humoral immunity and pathology of lymphoid organs. It is concluded that imidacloprid insecticide has immunological deleterious effects in chickens targeting the humoral immune responses and vitamin E and selenium supplementation decreased the ill effects of imidacloprid.

Clinicopathological studies on vitamin D 3 toxicity and therapeutic evaluation of Aloe vera in rats

A study was conducted to examine the clinical signs, hematological, biochemical and histopathological changes in vitamin D 3 toxicity at a dose rate 2 mg/kg b.wt. of vitamin D 3 and to assess the protective effect of Aloe vera in vitamin D 3 toxicity. The clinical signs observed were anorexia, progressive weight loss, difficulty in movement and respiration, diarrhea, epistaxis, subnormal body temperature and nervous signs before death. Mortality was observed in treated rats between day 10 and day 19 of treatment. The gross postmortem changes observed were severe emaciation, white chalky deposits on epicardial surface of heart, pin point white deposits on cortical surface of kidneys with pale yellow discoloration and diffused white deposits on serosal surface of stomach and intestine with bloody ingesta in lumen. The hematological changes included non-significant increase in hemoglobin and total leukocyte count and significant increase in relative neutrophil count. The biochemical changes observed were significant increase in plasma concentration of calcium, phosphorus and blood urea nitrogen, whereas a significant decrease in the concentration of albumin and total plasma protein was observed. The histopathological lesions included calcification of various organs, viz., tongue, stomach, intestines, kidney, heart, aorta, larynx, trachea, lungs, spleen, choroid plexus arteries of brain and vas deferens. The Aloe vera juice (2.5% in drinking water) has no protective effect on vitamin D 3 toxicity (2 mg/kg b.wt.).

Neuropathological studies of chickens following exposure to chlorpyrifos

The objective of this study was to determine the putative neuropathological effects in young chickens after administration of a single dose of 55 mg/kg bw chlorpyrifos. The gross lesions of the nervous system comprised of congestions in the brain. Microscopic examination of brain showed mild congestion of cerebral blood vessels and mild perivascular cuffing of lymphomononuclear cells in the cerebral cortex and necrosis of the neurons. The interesting findings were the presence of cytoplasmic vacuolations of cerebral neurons and swelling of the endothelial cell of the cerebral capillaries. Cerebellum showed congestion and hemorrhages in the granular layer and necrosis of Purkinje cell. Sciatic nerve exhibited mild edema, swelling and degeneration of axons, and swelling of Schwann cells. There was a significant inhibition of plasma cholinesterase enzyme activity in chickens administered with chlorpyrifos compared to chickens of control group. The study revealed that administration of chlorpyrifos produces neuropathological lesions in chickens shortly after exposure.

Histopathological and immunohistochemical diagnosis of infectious bursal disease in poultry birds

Aim: The aim of the present study was to diagnose infectious bursal disease (IBD) using gross, histopathological, and immunopathological approaches and to compare efficacy of immunohistochemical techniques with conventional diagnostic techniques. Materials and Methods: A total of 33 samples were collected from the six different poultry farms from Ludhiana and the nearby districts. Upon gross analysis of the necropsied birds, the relevant tissue samples such as bursa, kidney, junction of proventriculus and gizzard, heart, and muscles were then processed for histopathological and immunohistochemical studies. Results: Varied macroscopic changes were noted in bursa, characterized as swollen, hemorrhages to atrophy in size. Nonetheless, hemorrhages over thigh muscles were rarely seen. Histologically, the bursa showed prominent fibrotic and atrophic changes. Rarefaction of bursal follicles with intermittent infiltration of lympho-mononuclear cells with chronic cystic changes was additional changes, considered to be paramount for IBD. Expression and localization of IBD specific viral antigens were noticed mainly intracellular to the rarefied areas of bursal follicle section(s), in conjunction to inner lining of the cystic cavities of affected follicles. In addition, the junction of proventriculus and gizzard, the heart muscle, respiratory ciliated epithelium, and proventriculus also revealed positive expression to IBD virus (IBDV) antigen. Advanced immunopathological techniques, i.e., immunofluorescence further testified the evidence of antigen as positive green signal within affected follicles. Further consideration to the reliability of various techniques employed, positive correlation (r=0.64623) was emerged out with conventional pathological scoring. Conclusion: It is concluded that the bursa acts as an organ of choice for demonstrating IBDV antigen for specific diagnosis of disease using immunohistochemistry (IHC), and IHC staining is a precise, specific, rapid, and reliable method to demonstrate the IBDV antigen in the altered tissues due to IBDV infection.

Histopathological and immunohistochemical approaches for the diagnosis of Pasteurellosis in swine population of Punjab.

Aim: Infectious porcine bronchopneumonia, caused by Pasteurella multocida, is a widespread disease of major economic significance. Thus, the aim of the present study was to diagnose swine Pasteurellosis using gross, histopathological, and immunopathological approaches in the swine population of Punjab and to compare the efficacy of immunohistochemical (IHC) techniques with conventional diagnostic techniques. Materials and Methods: A total of 71 adult swine lung samples showing gross pneumonic changes were collected along with the associated lymph nodes to carry out the study. The collected samples were then processed for histopathological and IHC studies. Results: Out of the total 71 lung samples, 26 samples were found to be suspected for Pasteurellosis as per the microscopic changes observed, and out of these 26 samples, 16 cases were confirmed to be positive for Pasteurellosis by IHC. Varied macroscopic changes noted in lungs were pneumonic patches with consolidation of many lobes, congestion, and focal hemorrhages. Main lesions associated with lymph nodes were its enlargement and hemorrhages. Histologically, the lung showed fibrinous and suppurative bronchopneumonia, multifocal suppuration, thickening of septa with fibrin combined with cellular infiltration and edema. The higher IHC expression of P. multocida was seen in the bronchial epithelium besides in alveolar and bronchial exudate. Moreover, on comparing the histopathological and IHC scores which were calculated on the basis of characteristic microscopic lesions and number of antigen positive cells, respectively, a significant positive correlation (r=0.4234) was found. Conclusion: It was concluded that swine population of Punjab is having P. multocida infection. The gross and histopathological lesions can be helpful in the preliminary diagnosis of Pasteurellosis but needs to be supplemented by other immunodiagnostic tests. Moreover, IHC technique proved to be a specific, reliable, precise, and rapid technique to supplement these conventional methods of diagnosis for Pasteurellosis.

Localization of classical swine fever virus antigen in young piglets by immunohistochemistry

The present study investigated the diagnosis of classical swine fever in piglets of four to six months using gross lesions, histopathology and immunohistochemistry. Gross examination revealed hemorrhagic kidneys, lymph nodes and spleen along with button shaped ulcer in intestine. Histopathological examination revealed focal to diffuse haemorrhages in the red pulp of spleen with depletion of the lymphocytes and multiple ulcerative lesions with necrosis of the epithelial mucosa in large intestine. Immunohistochemically, the viral antigen was demonstrated in the bronchial epithelial cells, mononuclear cells in spleen, intestine and Purkinje cells of the brain.

Concomitant Effect of Chlorpyrifos and Intranasal Endotoxin Administration on Apoptosis Related Protein Expression in Lung of Mice

Pesticides and bacterial endotoxin have been implicated in variety of respiratory dysfunctions. In present study, mice were administered chlorpyrifos at 3 mg/Kg BW/day for 30 days (by oral gavage) and/or challenged with bacterial endotoxin at 80 μg/mice by intranasal route to investigate expression of apoptosis related proteins such as p53, Bax and caspase-3 in lung. The results of immunohistochemistry of lung showed significant increase in number of p53 expressing cells in chlorpyrifos treated mice as compared to control while endotoxin and chlorpyrifos+endotoxin treated mice showed increased number of p53 expressing cells as compared to control and chlorpyrifos treated mice. Chlorpyrifos or endotoxin administration resulted in increased expression of Bax and Caspase-3 while chlorpyrifos treated and endotoxin challenged mice showed synergistic effect on expression of these proteins. From these results, it is concluded that chlorpyrifos and/or endotoxin increased critical apoptosis related proteins such as p53, Bax and Caspase-3 in lung of mice, which may play pivotal role in pathophysiology of pulmonary dysfunctions resulting from exposure to pesticides and endotoxin.

Bromadiolone Induced Oxidative Stress and Cytological Damage in Layer Birds

This study was undertaken to evaluate the oxidative stress induced by bromadiolone toxicity in layers. Birds were divided into two groups. Control group (I) received bromadiolone free feed and group (II) was exposed to bromadiolone (5 mg/kg feed) for seven weeks. Blood samples and organs were collected at the end of the experiment. Bromadiolone exposure resulted in significant rise in lipid peroxidation (LPO) levels in erythrocyte and liver. In addition decreased ( P <0.05) level of reduced glutathione (GSH), superoxide dismutase (SOD), and catalase (CAT) activities were observed in erythrocyte and liver. Histopathological results revealed mild to severe type of necrosis and degenerative changes in heart and liver of bromadiolone intoxicated animals. In addition histopathological findings of tissues ovary, liver, heart and pancreas also showed generalized haemorrhage. It is therefore concluded that feeding of bromadiolone admixed feed lead to depletion of Vitamin K causing multifocal haemorrhages and oxidative stress and cellular damage in different tissues.

Diagnosis of Newcastle disease in broiler by histopathology and immunohistochemistry

Newcastle disease (ND) is an OIE notifiable, highly contagious, generalized viral disease of poultry. It remains a constant threat to the poultry industry and is a limiting disease for poultry producers worldwide. The present study report a severe enteric form of ND virus infections in backyard poultry birds by using histopathology and immunohistochemistry. ND viral antigens were detected in the lesions of various organs. ND viral antigen were mainly localized in the cytoplasm and nuclei of necrotic cells of epithelium of proventriculus, gizzard and intestines, Kupffer cells and circulating monocytes in the liver, tracheal epithelial cells, bronchiolar and parabronchiolar epithelial cells and alveolar macrophages in the lungs. Thus, immunohistochemistry serve as rapid and reliable tools to study the tropism and distribution of ND viral antigen in the tissues of infected birds and hence its diagnosis.

Epidemiological and immunopathological studies on Porcine parvovirus infection in Punjab

Aim: The aim of this study was to get the first-hand knowledge about the seroprevalence of Porcine parvovirus (PPV) in Punjab and a diagnosis of PPV from abortion cases of swine using gross, histopathological, and immunohistopathological techniques to observe the tissue tropism of the virus strain. Materials and Methods: Tissue samples from the reproductive tract of pig (n=32), placental tissue (n=10), and aborted fetuses (n=18) were collected from Postmortem Hall of the Department of Veterinary Pathology, GADVASU, field outbreaks and from butcher houses in and around Ludhiana. These samples were processed for histopathological and immunohistochemical (IHC) studies. For seroprevalence study, 90 serum samples of different sex and age were collected from 15 swine farms of Punjab and were subjected to indirect enzyme linked immunosorbent assay using commercial kit. Results: Overall, seroprevalence of PPV was found to be 41.1%. Sex and age related difference in the prevalence was noted. In abortion cases grossly congested and emphysematous lungs, congested internal organs with fluid in abdominal cavity and congestion in brain, changes were noted in fetuses, while diffuse hemorrhages and edema was observed in placental tissue. Histopathologically, the most frequent fetal lesions in aborted fetuses were noted in lungs, liver, and brain. IHC staining revealed PPV antigens in sections of heart, liver, lung, spleen, brain, lymph node of fetuses, placenta, and uterus of sow. Gross, histopathological, and IHC examination of the samples confirmed 5 fetus, 2 placenta and 3 female reproductive samples positive for parvovirus infection. Conclusions: Seroprevalence results may serve as a support either in prevention or control of the disease. IHC is the sensitive technique for diagnosis of PPV associated with the reproductive tract of swine and was found to supplement the gross and histopathological alterations, respectively, associated with the disease.