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Septicaemic diseases are the leading cause of mortality in piglets causing great economic loss to the porcine industry. Haemophilus parasuis, an important pathogen in piglets, is widely distributed in all pig rearing countries. Carrier pigs are considered as the major reservoir of H. parasuis and could be a significant source of infection leads to septicaemia and pneumonia in pigs. In this study, a total of 51 tissue samples from different outbreaks of septicaemia in and around Bareilly district of Uttar Pradesh state of India were evaluated for H. parasuis infection by PCR and histopathological examination. A total of 12 samples were found positive for H. parasuis on screening (12/51, 23.52%). On post mortem examination, the gross lesion in lungs, heart, spleen and brain indicated establishment of septicaemia and pneumonia. The predominant histological changes observed were fibrinous polyserositis and bronchopneumonia as compared to other tissue lesions such as splenitis, nephritis, myocarditis and meningitis. The involvement of lymph nodes and joints were also observed as an important finding of the H. parasuis infection. The study revealed H. parasuis is prevalent in Indian pigs and is responsible for large outbreaks of septicaemia. In this context, more studies targeting the H. parasuis are highly warranted to establish its pathology and pathogenesis among Indian pigs.

To understand the mechanism of host-pathogen interaction during the progression of Pasteurella multocida capsular serotype D of porcine origin causing pneumonia/septicemia, 36 adult Swiss albino mice were intranasally inoculated with 25 μl of P. multocida serotype D@109 CFU/ml and sacrificed at 6, 12, 24, 48, 72 and 96 hours post inoculation. The infected mice showed varied clinical signs of depression, ruffled fur, ocular discharge, watery to mucoid nasal discharge with labored breathing pronounced during 24-48 HPI and declined thereafter. The leucocytosis, anaemia, low PCV, progressive neutrophilia with lymphocytopenia changes in these infected groups of mice paralleled with the clinical signs. The serum biochemical parameters such as total protein, blood urea nitrogen, creatinine, glucose, SGOT, SGPT and C-reactive protein were significantly increased. The higher level of LPO, lower level of antioxidant enzymes (CAT, SOD and GSH) and higher values of proinflammatory cytokine genes (TNF-α, IL-1β and IL-6) in lungs and spleen of the inoculated mice as compared to control mice suggest towards the establishment of pneumonia and sep-ticemia. The lesions of bronchopneumonia, splenitis, nephritis, hepatitis, enteritis and myocarditis along with the demonstration of bacterial antigens in lungs and spleen well correlated with severity of histopathological lesions at different time points. Apoptotic nuclei were detected in infiltrating leukocytes of the lungs and splenocytes on TUNEL staining. The inflammatory lesions as well as the apoptosis were highest during 24-48 HPI followed by gradual decrease till the end of the experiment. This is the first report on elucidating the pathogenesis of P. multocida capsular serotype D of porcine origin.

A 4-week-old male crossbred piglet was presented for necropsy examination with history of neurological signs progressing to death. Grossly, the meninges covering the brain were thickened, opaque containing purulent exudates. Besides, the other visceral organs like lungs, heart, spleen, lymph nodes, kidneys and intestine were severely congested. Histologically, suppurative meningoencephalitis was observed in addition to suppurative bronchopneumonia, hepatitis, nephritis and splenitis indicating the septicemia in pig. Immunolabelling of S. suis antigen was observed in meninges, brain, lungs, spleen, tonsil and lymph nodes. Both S. suis and S. dysgalactiae subspecies equisimilis were isolated from the brain, lungs and spleen. The biochemical characterization by API 20 strep kit and genomic detection test by PCR further confirmed both the isolates. This is the first report of dual infection of S. suis and S. dysgalactiae subspecies equisimilis infection in a pig with severe disseminated suppurative meningoencephalomyelitis.

Aim: The small ruminant lentiviruses are known to cause maedi-visna (MV) and caprine arthritis - encephalitis in sheep and goats, typically affecting joints, udder, lungs, and the central nervous system. The diagnosis usually involves serology, clinical signs, immunohistochemistry, and polymerase chain reaction (PCR). In the present study, the histopathologically positive pneumonia cases of MV were confirmed by PCR in lung tissue probably for the first time in India. Materials and Methods: A total of 888 lungs of adult sheep, aged between 2 and 5 years, were screened during slaughter, of which 121 were found to have pneumonic lesions. The tissues from each pneumonic lung including associated lymph nodes were collected in 10% neutral buffered formalin for histopathology. The frozen tissues of the same were also collected and stored at -20°C for PCR confirmation. Results: Three of 121 cases of pneumonic lungs of sheep revealed gross and histopathological lesions suggestive of maedi or ovine progressive pneumonia infection. These 3 cases were further confirmed by PCR technique that amplified 291-base pair DNA in the long terminal repeat sequence of MV provirus. Conclusion: This study suggests the low occurrence of MV virus (MVV) infection in India in naturally affected sheep based on pathomorphological lesions and using the molecular tool of PCR detection of the virus in tissues. Further, a combination of pathomorphology or/and PCR testing might be optimal for detecting the animals infected with MVV.

Pasteurella multocida is an economically important respiratory pathogen of pigs confronting swine industry worldwide. Despite extensive research over the decades, its pathogenesis is still poorly understood. Recent reports have demonstrated the nervous system affection as a newer aspect of pathogenesis by Pasteurella multocida type B:2 in Haemorrhagic Septicemia, but there are no reports of the involvement of nervous system by P. multocida in pigs. Therefore, the study was aimed to explore the neurovirulence of Pasteurella multocida in naturally infected pigs. A total of 15 brains were collected from the natural cases of pig mortality suggestive of Pasteurellosis. Grossly, the leptomeninges were markedly congested and brains were oedematously swollen. Histologically, there was moderate to severe fibrinohaemorrhagic and mononuclear cells exudates present in the leptomeningeal tissue and cerebrospinal spaces. Similar vascular inflammatory lesions (perivascular and perineuronal) along with gliosis, neuronal degeneration and necrosis were noted in various subanatomical sites of the brain (cerebrum, cerebellum, brainstem and spinal cord). The culture and biochemical tests showed the presence of P. multocida within the brain tissue. P. multocida type specific antibody staining in the brain tissues revealed intense distribution of antigens in the inflammatory exudates of meningeal vessels, neurons, glial cells and endothelial cells of the blood vessels contributing its association with neuropathological lesions. Pasteurella multocida specific PCR amplification of capsular polysaccharide gene yielded 460 bp and multiplex PCR showed the involvement of capsular serogroups A &D. All the isolates showed the presence of 10 genes for virulence factors. The disease confirmation of both serotypes was proven by Koch's postulates using Swiss albino mice. Further, histopathological brain lesions along with the immunohistochemical detection of bacterial antigens were corroborated with natural cases of P. multocida as described above. To the best of our knowledge, we first time report the neuroinvasion of P. multocida in naturally infected pigs.

Peste des petits ruminant (PPR), a highly contagious viral disease of small ruminants, is characterized by erosive stomatitis and pneumo-enteritis. However, its neurovirulence potential as observed with other morbilliviruses has not been fully investigated. The present study describes the neuropathological alterations induced by PPR virus through apoptotic pathway. A total number of 12 carcasses of local breed goat kids of either sex were received for postmortem examination. The clinical history was described as symptoms of mucopurulent nasal discharge, high to low grade fever, erosive stomatitis, dyspnoea and profuse watery diarrhoea followed by mortality of 35 goat kids within a week. The pathoanatomical lesions and immunohistochemical demonstration of PPRV antigen in lungs, intestine, spleen and lymph nodes confirmed PPR disease in goats. Grossly, five brain specimens showed moderate to severe leptomeningeal congestion during necropsy. Microscopically, brain sections showed leptomeningitis and nonsuppurative encephalitis characterized by vascular congestion, haemorrhages in the parenchyma, perivascular cuffing with mild to moderate mononuclear cells (mainly lymphocytes and few macrophages), focal to diffuse microgliosis, neuronal degeneration, satellitosis and neuronophagia. Immunolabelling of viral antigen was observed in the cytoplasm of neurons and glial cells. The RT-PCR amplification of N gene fragment also confirmed the presence of PPRV in the brain. The strong immunoreactivity of Caspase-3, Caspase-8 and comparatively lower expression of caspase-9 along with the absence of any reactivity for Apaf-1 antigen in the brain sections indicated the role of caspase dependent extrinsic pathway in inducing neuropathological changes. The presence of apoptotic neurons in the brain by TUNEL assay further confirmed the apoptosis and strong immunoreactivity of iNOS in neurons which suggested the generation of oxidative stress, that might have induced the apoptosis. The overall findings confirm the neurovirulence potential of PPR virus, via the extrinsic pathway of apoptosis, in natural cases of PPR disease in goat kids.

Kaempferol is a flavonoid and important part of the diet. Kaempferol has shown antioxidant, antiinflammatory and antidiabetic activities in various studies. However, protective potential of kaempferol in acute lung injury induced by sepsis and its mechanism remains unclear. The present study was undertaken to evaluate the effect of kaempferol in sepsis-induced acute lung injury in mice and its possible mechanism of action.Acute lung injury was induced by CLP surgery in mice. Kaempferol (100 mg/kg bw) was administered orally one hour before caecal ligation and puncture surgery in mice. Mice were divided into four groups sham, KEM+sham, sepsis (CLP), and KEM+sepsis. Assessment of lung injury was done by estimation of protein content in lung tissue, lung edema, proinflammatory cytokines in plasma and lung tissue, oxidative stress, antioxidant enzymes, nitrite production, and histopathology.Kaempferol pretreated mice showed significant (P0.001) decrease in water content in lungs. Kaempferol pretreatment showed reduction in cytokines IL-6, IL-1β, and TNF-α in plasma as well as in lung tissue in comparison with septic mice without pretreatment. Pretreatment with kaempferol did not show any reduction in MDA level in comparison with septic mice. Antioxidant enzymes SOD and catalase and nonenzymatic antioxidant GSH activities were also increased with kaempferol pretreatment in septic mice. Further, kaempferol pretreatment reduced the lung tissue nitrite level (P0.01) and iNOS (P0.05) level in septic mice. A significant (P0.01) downregulation of mRNA expression of ICAM-1 and iNOS was observed with this pretreatment. Kaempferol pretreatment did not decrease bacterial load in septic mice. Mice pretreated with kaempferol followed by sepsis showed lesser infiltration of cells and more arranged alveolar structure in histopathological analysis.The study suggests that kaempferol showed attenuation in sepsis-induced acute lung injury in mice through suppression of oxidative stress, iNOS, and ICAM-1 pathways.

Xanthium strumarium has traditionally been used in the treatment of urolitiasis especially by the rural people in India, but its antiurolithiatic efficacy was not explored scientifically till now. Therefore, the present study was designed to validate the ethnic practice scientifically, and explore the possible antiurolithiatic effect to rationalize its medicinal use. Urolitiasis was induced in hyperoxaluric rat model by giving 0.75% ethylene glycol (EG) for 28days along with 1% ammonium chloride (AC) for first 14days. Antiurolithiatic effect of aqueous-ethanol extract of Xanthium strumarium bur (xanthium) was evaluated based on urine and serum biochemistry, oxidative/nitrosative stress indices, histopathology, kidney calcium and calcium oxalate content and immunohistochemical expression of matrix glycoprotein, osteopontin (OPN). Administration of EG and AC resulted in hyperoxaluria, crystalluria, hypocalciuria, polyurea, raised serum urea, creatinine, erythrocytic lipid peroxidise and nitric oxide, kidney calcium content as well as crystal deposition in kidney section in lithiatic group rats. However, xanthium treatment significantly restored the impairment in above kidney function test as that of standard treatment, cystone. The up-regulation of OPN was also significantly decreased after xanthium treatment. The present findings demonstrate the curative efficacy of xanthium in ethylene glycol induced urolithiasis, possibly mediated through inhibition of various pathways involved in renal calcium oxalate formation, antioxidant property and down regulation of matrix glycoprotein, OPN. Therefore, future studies may be established to evaluate its efficacy and safety for clinical use.

Complete cDNA sequences of kiss1 ( gmkiss1 ) and its receptor kiss1r ( gmkiss1r ) were cloned and characterized from brain tissue of adult golden mahseer ( Tor putitora ). Thereafter, quantification of gmkiss1 and gmkiss1r mRNA expression in brain-pituitary-gonad (BPG) axis of male and female golden mahseer was carried out using quantitative real-time (qRT)-PCR assay during an annual reproductive cycle, at different gonadal development stages. The gmkiss1 cDNA was 508 bp, with 330 bp open reading frame (ORF), encoding a precursor protein of 109 amino acids, whereas gmkiss1r cDNA was 1383 bp with an ORF of 1004 bp, which encodes a 334 amino acid protein residue. The qRT-PCR study shows that gmkiss1 and gmkiss1r are expressed in brain, pituitary and gonads of both the sexes of golden mahseer. An apparent sexual dimorphism in transcript level of gmkiss1 and gmkiss1r in brain and gonads was evident during the reproductive cycle. Overall, in brain, testis and ovary, the gmkiss1 and gmkiss1r mRNA expression level was comparatively higher during the initial stages of gonadal development, than that of spermiation or ovulation stage. In pituitary of both the sexes, throughout the gonadal development, consistently low transcript level of gmkiss1 and gmkiss1r was observed. The gmkiss1 mRNA expression level in brain and ovary of female golden mahseer was several folds higher than the brain and testis of male fish. In conclusion, we confirm the presence of kiss1 and its receptor in golden mahseer, and results of our study strongly suggested the involvement of kisspeptin1 system in gonadal development and annual reproductive cycle of this species.

Microcystins (MCYST) are the freshwater cyanobacterial toxins, known to induce hepatocellular carcinoma, necrosis, intrahepatic bleeding, as well as human and livestock mortality. Within hepatocytes, MCYST selectively bind to protein phosphatases 1 and 2A, resulting in severe liver damage. The toxicology of MCYST in mice and rats has been well studied, but little is known regarding genotoxicity in aquatic animals. In this study, the zebrafish, Danio rerio was exposed to crude extract of Microcystis aeruginosa bloom. Liver and heart were examined for MCYST-induced toxicity. Light microscopy at 36 h revealed severe, widespread apoptotic necrosis of the majority of hepatocytes, and cytoskeletal deformation in myocardiocytes. Hepatocytes were dissociated with cell shrinkage and margination of nuclear chromatin. Laddering of genomic DNA from the liver and heart of the exposed fish in an increment of 180–200 bp was consistent with apoptosis. Fluorimetric analysis of DNA unwinding was carried out to determine th...

Hepato-renal syndrome is a type of functional kidney failure due to acute or chronic liver diseases. A five year old female, crossbred goat was presented for post-mortem examination. On necropsy, icteric discoloration of carcass with enlarged liver, thickened jejunal and ileal mucosa and enlarged mesenteric lymph nodes were observed. Severe lipidosis of liver, hypertrophied hepatocytes, mononuclear cell infiltration of parenchyma and portal triads, fusion of villi, and extensive infiltration by mononuclear cells (epithelioid-macrophages, plasma cells and lymphocytes) and occasional giant cells were observed around a central caseated and calcified mass surrounded by fibrous tissue proliferation in the lamina propria of intestine. Calcification of the mesenteric lymph node was confirmed by Von Kossa staining as dark brown to black deposits. Severe glomerular and tubular degeneration, necrosis and presence of bile casts were observed in kidney, which was confirmed by Hall's staining. Diffuse golden yellow hemosiderin pigment deposits were observed in the splenic parenchyma, which wasconfirmedbyPerl's Prussianbluestainingasbluishblackpigments. Ziehl-Neelsenstainingrevealedacid-fastbacilli inthelamina propria of intestine. Immunohistochemistry (IHC) using anti-M. aviumsubspp. paratuberculosis immune serum showed positive brown signals intracellularly in the macrophages and epithelioid cells in the lamina propria of intestine. Demonstration of acid-fast organism and IHC confirmed paratuberculosisinfection and the cascade of injury consequent to the disease might have resulted in hepato-renal syndrome and death of the affected animal.

Altogether 185 pig carcasses from privately run slaughter slabs of Rohilkhand region were screened to ascertain the prevalence of cysticercosis. Out of them, 14 cases (7.56%) were found to be infected with cysticercosis. The cysts were found distributed heavily within the skeletal, cardiac and scrotal muscles, and onto the leptomeninges of the brain causing pressure atrophy. Histological examination showed cystic structures within the musculature along with vascular congestion, infiltration of inflammatory cells within the connective tissue. The brains showed cystic cavities with or without the presence of parasites, meningo-encephalitis, perivascular cuffing, neuronophagia and focal gliosis.

Corneal hypertrophy was observed in fingerlings of chocolate mahseer, Neolissochilus hexagonolepis (McClelland, 1839) from Uttarakhand, India. The advanced stage of infection was marked with damage of cornea and shrunken eye. Skeletal deformity was also recorded in some of the infected fishes. Samples of eye and other organ were collected for histopathological, parasite and microbial investigation. Histological examination of the eye indicated ocular collapse, entirely damaged cornea, disruption of vitreous, aphakic globe, oedema of choroid and optic nerve and, deformed retina. Microscopic observations of wet mounts and histopathology ruled out infectious agents like microsporidia, myxosporidea, cystercoidiasis, atypical mycobacteria, digeneans and fungi. PCR for viral aetiology did not yield any significant amplicons. Further detail study related to physic-chemical parameter need to be carried out to ascertain the exact cause of this disease.

A rare case of ‘Diprosopus monauchenos’ was recorded in a newborn live piglet from a five year-old multipara sow. The unique animal had two faces fused caudally by a single parietal bone and joined ventrally by the medial ramie of the mandible. The ‘diprosopus’ consisted of four eyes (tetraophthalmus), two pairs of nostrils, two mouths and each with a tongue. The piglet died on second day besides best nourishing care. Clinical and radiographic observations revealed normal anatomic, thoracic and abdominal viscera and confirmed the aforementioned findings.