Novel pathologic findings and viral antigen distribution in cattle and buffalo calves naturally infected with Foot-and-Mouth disease virus.

The Foot-and-Mouth disease is highly contagious acute viral disease of livestock inflicting huge economic loss to the farmers. The limited knowledge regarding the pathological lesions vis-a-vis distribution of the FMDV in lesser explored endocrine glands and important vital organs other than the target organs of infected calves prompted us to take the present investigation to have detailed insight into the pathogenesis. The systematic necropsy of 37 dead calves (cattle-28 and buffalo-9) was conducted, and thin representative tissue pieces from the affected organs were collected in 10% neutral buffered formalin (NBF) for pathological and immunohistochemical investigations. The genomic detection and its serotyping were done by RT-PCR and multiplex-PCR, respectively. Necropsy examination in all cases showed myocardial lesions resembling 'tigroid heart appearance'. Other organ specific lesions include vesiculo-ulcerative stomatitis, edema of the lungs, petechial hemorrhages, edema of the endocrines, and gastroenteritis. Histopathological examination showed varying sizes of vesicles and ulcerations in stratified squamous epithelium of the tongue, acute necrotizing myocarditis, lymphoid depletion in lymphoid tissues, hepatitis, pancreatitis, thymic hyperplasia, thyroiditis, adrenitis, and enteritis. Positive immunolabeling for viral antigens was observed in endocrine glands, lymphoid organs, lungs, liver, kidneys, and intestine, in addition to other typical locations. The thyroid, adrenal glands, and pancreas, in addition to the tongue and heart, are the tissue of choice for sampling in the field during epidemics. Further, the viral genome and serotype A was confirmed in the affected tissues. This study provides insights into novel tissue tropism and pathogenesis in young calves naturally infected with FMDV.

Patho-morphological and immunohistochemical diagnosis of fibrous tissue pulmonary hamartoma in young buffaloes.

Hamartoma is a developmental disorder characterized by the presence of well-differentiated tissue, of multiple types, in excess at its normal location. The present study describes a fibrous tissue pulmonary hamartoma in two buffaloes. Grossly, there were large pale white masses present on the left diaphragmatic lobe in both cases. Microscopic examination of the lungs revealed that the bronchioles were dilated and showed many terminal respiratory bronchioles lined by a single layer of cuboidal epithelium supported by thick bands of collagenous tissue. Alveoli were distorted and separated by wide bands of connective tissues. Interalveolar septa were greatly expanded by collagen fiber proliferation and subsequent fibrosis. Fibrous connective tissues were demonstrated by immunohistochemistry (IHC) and Masson's Trichrome staining (MTS). Based on gross and histological examination, coupled with IHC and MTS, it was possible to confirm a case of hamartoma, which is rarely reported worldwide and is the first reported case from India.

Type II Myocardial Infarction: Predisposing Factors, Precipitating Elements, and Outcomes.

Introduction Myocardial infarction (MI) is a subset of the spectrum of the disease known as acute coronary syndrome (ACS), which comprises three distinct entities including unstable angina (UA) and MI with or without ST-segment elevation. However, many clinicians are unaware that MI itself is classified into five types, the most common being type I, followed by type II. Type II MI occurs due to coronary insufficiency not related to acute plaque change in the coronary vasculature. Data available on type II MI is still limited, particularly in the South Asian setting, despite documented poorer outcomes for the same compared to other types. Therefore, we conducted this study as an attempt to outline the predisposing factors, precipitating elements, and possible outcomes of type II MI. Materials and methods This prospective study was conducted at a tertiary care hospital in Kochi, Kerala for 12 months. A total of 59 patients of ages 10-99 years, with a final diagnosis of MI based on the levels of cardiac biomarkers and electrocardiography (ECG), no previous history of coronary angiography, thrombolysis, percutaneous coronary intervention (PCI), and non-ischemic conditions producing elevations in cardiac biomarkers were included in this study. Demographic features, cardiac biomarker levels, comorbidities, precipitating factors, foci of sepsis, and outcomes of type II MI were noted. The mean was calculated for age and cardiac biomarkers. The frequency and percentages were calculated for gender, comorbidities, precipitating factors, foci of sepsis, and the outcomes of type II MI. Results The mean age of the patients was 69.66 years; 38 (64.4%) patients were males and 21 (35.59%) were females. Mean elevation of creatine kinase myocardial band (CK-MB) was 47.457 IU/L and highly sensitive troponin I (Hs-Trop I) was 8.712 ng/mL. Diabetes mellitus [44 (74.57%)] and hypertension [41 (69.49%)] were the most common underlying patient comorbidities followed by dyslipidemia [38 (64.4%)]. Most of the patients had more than two comorbidities at a time; 33 (55.93%) patients had sepsis, 31 (52.4%) patients had anemia, 29 (49.1%) patients had electrolyte imbalance, 19 (32.2%) patients had respiratory failure, 16 (27.11%) patients had arrhythmia (tachyarrhythmia/bradyarrhythmia), and two (3.3%) patients had postoperative (non-cardiothoracic) stress. Sepsis was originating from the lower respiratory tract in 14 (42.42%) patients, blood in 11 (33.33%) patients, urinary tract in eight (24.24%) patients, and abdomen in six (18.18%) patients. Thirty-four (57.62%) patients had heart failure, 13 (22.03%) had arrhythmias, and 19 (32.20%) patients died. Conclusions Type II MI has a high mortality rate, mostly due to heart failure and arrhythmia. Patients with diabetes mellitus and hypertension are at increased risk of type II MI. Sepsis is the most common precipitating factor, primarily originating from the lower respiratory tract, followed by anemia and dyselectrolytemia. Treatment of precipitating factors is the primary way to manage type II MI.

Immunohistochemical and molecular detection of natural cases of bovine rotavirus and coronavirus infection causing enteritis in dairy calves.

Bovine rotavirus (BRoV) and bovine coronavirus (BCoV) are major enteric viral pathogens responsible for calve diarrhoea. They are widespread both in dairy and beef cattle throughout the world and causing huge economic losses. The diagnosis of these agents is very difficult due to non-specific nature of lesions and the involvement of some intrinsic and extrinsic risk factors. We performed postmortem of 45 calves, which was below three months of age. Out of 45 necropscid calves, three (6.66%) cases were positive for BRoV and four (8.88%) cases were found positive for BCoV, screened by reverse transcriptase polymerase chain reaction (RT-PCR). Further RT-PCR positive cases were confirmed by immunohistochemistry (IHC) in paraffin-embedded intestinal tissue sections. Three cases of enteritis caused by BRoV showed the hallmark lesions of the shortening and fusion of villi, denudation and infiltration of mononuclear cells in the lamina propria. The BRoV antigen distribution was prominent within the lining epithelium of the villi, peyer's patches in the ileum and strong immunoreactions in the lymphocytes and some macrophages of the mesenteric lymph nodes. Four cases in which BCoV was detected, grossly lesions characterized by colonic mucosa covered with thick, fibrinous and diphtheritic membrane. Histopathologically, jejunum showed skipping lesion of micro-abscesses in crypts. The BCoV antigen distribution was prominent within the necrotic crypts in the jejunum and cryptic micro-abscesses in the colon and ileum. It is the first report of BRoV and BCoV antigen demonstration in the jejunum, colon, ileum, Peyer's patches and mesenteric lymph nodes of naturally infected calves from India by using IHC.

Experimental bovine rotavirus-A (RV-A)infection causes intestinal and extra-intestinal pathology in suckling mice.

We describe here the intestinal and extra-intestinal spread of the species A rotavirus (RV-A) and associated lesions thereof in Swiss albino suckling mice pups, inoculated with a bovine-origin RV-A strain. In total, 35 suckling pups were used, wherein 20 pups received cell culture isolated RV-A @ 160 µL (TCID50/ml, 5 × 106.5) per pup [oral 80 µL and intra peritoneal (IP) 80 µL] and served as an infected group, while 15 pups were kept in the control group and inoculated the same volume of phosphate buffered saline (PBS) of neutral pH orally and IP. Four pups from the infected group and 3 from control group were sacrificed at 3, 5, 7, 9 and 12 day post infection (DPI). Of note, infected pups exhibited signs of dullness and restlessness till 5DPI, but none showed diarrhea at any point of time. No appreciable gross lesions were evident in any of the organs, except for mild congestion of the small intestine and yellowish catarrhal smearing over the luminal surface. However, light microscopic lesions in hematoxylin and eosin (H&E) stained sections of jejunum and ileum revealed vacuolation and pyknosis of nuclei of the mature enterocytes, their lysis and detachment, constriction and detachment of villi, mild mononuclear cells (MNCs) infiltration in the lamina propria and mildcell depletion of Peyer's patches and mesenteric lymph nodes (MLN). The extra-intestinal lesions of the cellular degeneration and mild MNCs infiltration were identified in the liver and kidneys from 3 to 7 DPI, but no lesion was seen in the brain. Interstitial thickening with MNCs of lung parenchyma was visible from 3 to 7 DPI. The lesions in the intestine, lymphoid tissues and lungs resolved after 7 DPI. The presence of viral nucleic acid was seen in the intestinal contents from 3 to 5 DPI by using a RV-A specific reverse-transcription polymerase chain reaction (RT-PCR), while in the MLNs and the lungs it could be detected till 5 DPI by both the RT-PCR and direct fluorescent antigen test (dFAT). However, liver, spleen and brain were tested negative for the presence of RV-A by any of these tests. Nonetheless, the persistence of the RV-A was seen in the MLNs even after the absence of virus from the small intestines. Findings here conclusively indicates that heterologous host origin RV-A has an affinity not only to the intestine but also to extra-intestinal tissues like MLNs and lung tissues.

Pathology and polymerase chain reaction detection of ovine progressive pneumonia (maedi) cases in slaughtered sheep in India.

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.