Multidrug resistant staphylococci isolated from pigs with exudative epidermitis in North eastern Region of India.

Exudative epidermatitis or greasy pig disease (GPD) is a contagious disease of pig and endemic worldwide caused by toxigenic strains under genus Staphylococcus. The present study reported an outbreak of GPD in Champhai district of Mizoram adjoining to the southern border of Myanmar. A total of 60 samples were collected from 22 clinically affected animals and processed for isolation and identification of Staphylococcus spp. All the isolates were subjected to antimicrobial sensitivity assay, biofilm production assay and detection of virulence genes, biofilm genes and mec genes followed by cloning and sequencing for phylogenetic analysis. A total of 44 staphylococci belonged to four species (S. sciuri, S. aureus,S. lentus, and S. hyicus) were isolated. Majority of the isolates were multidrug resistant with maximum resistance against ampicillin, penicillin including vancomycin. None of the S. hyicus isolates was methicillin resistant (MRSH) but 66·67% isolates were MRSA. By PCR, mecA gene was detected in S. aureus (n = 2), S. sciuri (n = 4) and S. lentus (n = 3). Biofilm associated gene icaD was detected in S. aureus (n = 3), S. sciuri (n = 5), S. hyicus (n = 4) and S. lentus (n = 6). The exfoliative toxin genes (ehxB, shetA and tsst1) were detected in S. hyicus (n = 3) and S. aureus (n = 1) isolates. All the isolates were closely related with the isolates from pigs of China, Germany, Japan and USA. The pathogens might be transmitted through illegal migration of pigs from Myanmar to India.

Cytokine responses in pigs after natural infection with classical swine fever virus.

Classical swine fever (CSF) is a highly contagious viral infection that affects domestic and wild pig population. The classical swine fever virus (CSFV) targets immune cells which perturb the immune functions causing immunopathological disorders such as immunosuppression, leukopenia and haemorrhage. In the present study, ELISA and quantitative real-time reverse transcription PCR (qRT-PCR) analysis was employed to determine cytokine profiles in pigs naturally infected with CSFV using whole blood assay (WBA) under field conditions. Significantly higher TNF-α, IL-10, and IL-6 expression levels were found in unvaccinated pigs infected with CSFV (group B) compared to vaccinated pigs that recovered after CSF (group A), the difference being statistically significant (p = 0.001). However, the expression of IFN-γ was significantly higher in group A compared to group B (p = 0.001). The findings of this field-supported study will help us to understand the immune biology of CSFV infection in infected pigs. The WBA technique can be used as a reliable, fast and feasible in vitro method to assess porcine cellular immune responses as it imitates the porcine blood conditions. Such studies could be of some value in determining the immune status of the ailing animals infected with CSFV. Keywords: classical swine fever virus; immune response; field conditions; interleukin; IFN-γ; TNF-α.

Unusual rotavirus genotypes in humans and animals with acute diarrhoea in Northeast India.

Rotavirus (RV) infection causes acute infantile diarrhoea in humans and animals and remains a major concern for vaccine development. The close proximity of humans to animals may foster cross-species infection resulting in the emergence of novel/unusual strains by genetic reassortment. In this study, we characterized 500 diarrhoeal samples for group A rotaviruses (RVA) from children (n = 290), piglets (n = 95) and calves (n = 115) in Northeast India during 2012-2013. The data showed that 142/500 (28·4%) faecal samples were positive for RVA with the highest level of infection detected in piglets (57/142, 40·1%) followed by children (51/142, 35·9%) and calves (34/142, 23·9%). Sequence-based G- and P-typing showed G1P[8] (25%) and G1P[7] (35%) were the prevailing genotypes in both humans and animals. Single cases of unusual genotypes, i.e. G9P[8], G5P[8] in humans and G1P[13], G1P[23] and G3P[7] in animals were also identified. Cluster analyses of the sequences showed regional strains were genetically closer to their homologous strains. However, human G5P[8] and porcine G1P[8] strains showed homology to heterologous hosts of their prototype strains. The subsequent global spread of unusual RV strains may result in their establishment over time, presenting challenges to future vaccine evaluation programmes. More studies on emerging genotypes are required to elucidate how RVA strains evolve post-vaccination. This study supports the need for continuous surveillance of RVA infections after detecting from diverse hosts in a common setting.

Status of Serum Zinc in Multidrug Resistant Tuberculosis.

Zinc plays a vital role in the immune status. Its deficiency affects host defense by reducing the number of circulating T cells and phagocytosis activity of other cells which ultimately impair cell mediated immunity. The cell-mediated immunity plays a major role in the causation of pulmonary tuberculosis. The present study was carried out to estimate serum zinc level in newly detected multidrug resistant tuberculosis (MDR-TB) in adult population. In this study total fifty (50) MDR-TB patients were enrolled conveniently from the in-patients departments of National Institute of Diseases of the Chest Hospital (NIDCH), Bangladesh. Serum zinc was estimated by atomic absorption spectrophotometry method from early morning fasting blood sample. Serum zinc level was assessed according to normal cut-off value 70-120 µgm/dl and 76% studied population were found lower than this value. The mean±SD serum zinc level was observed 60.40±8.91 µgm/dl. No associations were found between serum zinc level with age (p=0.11) and with sex (p=0.085) of the study population respectively. The low level of serum zinc in MDR-TB patients suggested impaired immune status of our study population.

Biomass Fuel Smoke and Tuberculosis: A Case-Control Study.

This case control study was done to ascertain the association between exposures to biomass cooking fuel smoke and pulmonary tuberculosis. Cases were all newly diagnosed pulmonary tuberculosis patients, randomly collected from the Hospital (DOTS centers) Registry from January 2012 to June 2012 from two Upazillas of Sirajganj district, Bangladesh. A home visitation program was done subsequently. Healthy controls were recruited from the neighbourhood of each case through a predefined procedure. Total 276 new pulmonary tuberculosis cases and 276 neighbourhood controls were enrolled. A semi-structured questionnaire containing demographic information, smoking habits, cooking place, kitchen condition, use of biomass fuel for cooking was used for interview. Crude (unadjusted) odd ratios and their corresponding 95% confidence limits for independent variables were determined by binary logistic regression. All significant variables were entered into a multivariate conditional logistic regression model and a final fitted model was determined by backward elimination methods. In univariate analysis, member of a nuclear family {Unadjusted odd ratio (Unadj. OR) 1.570, 95% CI 1.008-2.44)}, having non-formal education (Unadj. OR 2.739, 95% CI 1.219, 6.153) or primary (Unadj. OR 3.407, 95% CI 1.492, 7.782) to secondary level (Unadj. OR 2.392, 95% CI 1.032, 5.544) education, using cow dung (Unadj. OR 3.961, 95% CI 1.267, 12.376) and biomass fuel i.e. plant origin (Unadj. OR 3.382, 95% CI 1.087, 10.518) for cooking, past smoker (Unadj. OR 2.504, 95% CI 1.061, 5.910), using open oven (Unadj. OR 3.109, 95% CI 0.995, 9.716), having small kitchen area (Unadj. OR 0.99, 95% CI 0.989, 0.999) all were significantly associated with tuberculosis. When all significant variables were entered into a multivariate conditional logistic regression model nuclear family {Adjusted odd ratio (Adj OR) 1.808, 95% CI 1.127, 2.9)}, primary level education (Adj OR 2.86, 95% CI 1.23, 6.647) and non formal education (Adj OR 2.312, 95% CI 1.013, 5.276), biomass fuel (plant origin, Adj OR 3.497, 95% CI 1.059, 11.55), and cow dung (Adj OR 4.368, 95% CI 1.319, 14.464)) used in cooking and irregular smoker (who stop smoking at least 6 month back) (Adj OR 2.806, 95% CI 1.141, 6.901) were significantly associated with tuberculosis cases. From this study significant association have been found between exposure to combustion of biomass cooking fuel and tuberculosis.

Comparison between Decompressive Craniectomy with Durotomy and Conservative Treatment in Spontaneous Supratentorial Intracerebral Hemorrhage.

Large Intracerebral hematoma (ICH), compounded by perihematomal edema can produce severe elevations of intracranial pressure (ICP). Decompressive craniectomy (DC) beneficially addresses mass effect. Therefore this study is aimed to prove that decompressive craniectomy with durotomy has better outcome in patients with spontaneous supratentorial ICH than conservatively treated patients. This Quasi-Experimental study was carried out in the Department of Neurosurgery, Dhaka Medical College & Hospital (DMCH), Dhaka, Bangladesh from July 2012 to December 2013 using purposive sampling procedure. A total of 80 admitted adult hypertensive patients with spontaneous supratentorial ICH were enrolled in the study population fulfilling all selection criteria. Of them 40 patients underwent decompressive craniectomy with durotomy was considered as surgical group and 40 patients were treated conservatively was considered as conservative group. All the cases were continuously followed up and GCS score at 7(th) day of treatment (D7) and at discharge was recorded. Mean±SD hematoma volume was 56.91±13.72ml in surgical patients and in conservative group 51.80±13.58ml. Outcome measured by modified Rankin Scale at 3 months. Sixty percent (60%) patients had good outcome (mRS 0-4) and 40% patients had poor outcome (mRS 5-6) in surgical group. On the other hand, 52.5% patients had good (mRS 0-4) and 47.5% had poor (mRS 5-6) outcome in conservative group. In logistic regression analysis, conservative group was 3.643 times more prone to develop poor outcome than surgical group which was statistically significant (95% CI, 1.040-13.047; p value <0.05) and volume of hematoma [OR (95% CI), 1.131(1.059-1.207); p value <0.001)] was the most important predictor of outcome. This study indicates that decompressive craniectomy with preservation of brain integrity in patients with spontaneous supratentorial ICH is feasible and safe. It can be a useful alternative surgical procedure in the treatment of spontaneous supratentorial ICH.

Restriction fragment length polymorphism analysis of rotavirus VP7-encoding gene from humans and animals of Northeast India: a relative study of Indian and global isolates.

A restriction fragment length polymorphism (RFLP) assay was developed to examine the genetic relationship between 67 (29 Indian, 38 global) rotavirus isolates of human, bovine and porcine neonates. The assay involved direct digestion of RT-PCR amplified VP7 cDNAs with three restriction enzymes (VspI, HaeIII, NlaIV) independently. Forty-eight RFLP patterns were identified for all 67 strains, and of these 20 patterns were associated with Indian isolates. A correlation between the restriction patterns and G type was apparent through deduction of enzyme restriction sites from known sequences. Major G serotypes (G1, G2, G6, G8) with a few mixed types could be differentiated where there was a positive assortment of intrinsic serotypes from multiple host origin, and certain single or combined enzyme profiles were highly dominant in the population. Significant genetic variations were established between global and Indian isolates and none of the RFLP patterns were shared between them. These data suggest that the Indian wild-type rotavirus population is distinguishable based on the VP7 gene, and co-circulation of distinct strains in different hosts is foremost, indicating the possible likelihood of inter-species transmission.

Gender Variation in the Risk Factors with Ischemic Stroke: Bangladesh Perspective.

It is suggested in different studies that gender has a great impact on the distribution of risk factors of stroke. This study aims at determining the gender differences in risk factors of acute ischemic stroke among male and female patients. Data extracted for this study were taken from a case control study which was carried out in the departments of Neurology and Medicine of Dhaka Medical College Hospital (DMCH) and Shaheed Suhrawardy Medical College Hospital (ShSMCH) Dhaka from July 2009 to June 2011. We collected data of 125 patients with ischemic stroke and 129 healthy controls from the primary source. All secondary data were divided on the basis of gender. Among all male respondents 70 stroke patients were considered as case and 72 healthy respondents were considered as control and in female group 55 were stroke patients (case) and 57 were healthy control. Risk factors of stroke for male and female were evaluated separately. After multivariate analysis ever smoking (both present and ex smoker) (adj. OR 3.799, 95% CI 1.416, 10.19), diabetes mellitus (adj. OR 5.687, 95% CI 1.99, 16.26), hypertension (adj. OR 18.53, 95% CI 6.43, 53.4) and total cholesterol (adj. OR 1.017, 95% CI 1.01, 1.03) were found significant predictors for stroke in male whereas diabetes mellitus (adj. OR 6.989, 95% CI 1.54, 31.64), hypertension (adj. OR 17.85, 95% CI 5.1, 62.38), serum low density lipoprotein (LDL) (adj. OR 1.04, 95% CI 1.013, 1.07) and serum triglyceride (TG) (adj. OR 1.012, 95% CI 1.004, 1.02) were found significant in female. Besides taking the controlling measure for hypertension and diabetes mellitus, life style modification is an important factor to prevent stroke in both sexes.

Classical swine fever in the pygmy hog.

The pygmy hog is a rare, small and highly endangered mammal belonging to the Suidae family, and it is presently found only in the Assam state of India. While investigating the cause of death of pygmy hogs housed at a conservation centre for captive breeding and research at Basistha, Assam, it was confirmed that they were susceptible to and died as a result of contracting classical swine fever (CSF), caused by CSF virus (CSFV), which is a highly infectious endemic disease of domestic pigs in India. The post-mortem findings and serum CSFV-specific antibody titres, along with the isolation of CSFV from two pygmy hogs, and further confirmation by CSFV genomic E2 and 5' untranslated region (UTR) gene amplification in PCR (polymerase chain reaction), clearly established the cause of death of the pygmy hogs. Further, on phylogenetic analysis, the pygmy hog CSFV 5' UTR sequences were grouped in the genotype 1.1 cluster of Indian CSFVs, and hence the strains causing infection were closely related to CSFV isolates circulating in domestic pigs. Therefore, the occurrence of CSF in this endangered species may pose a potent threat to their existence unless properly controlled, and thus it needs urgent attention. To the authors' knowledge this is the first report on CSF in pygmy hogs.

Lymphocyte subsets in the small intestine of piglets fed with probiotic and zinc: a qualitative and quantitative micro-anatomical study.

BACKGROUND: Piglet mortality is a real concern to the pig farmers. The major cause is due to the late maturation of the immune system and dietary changes in postweaned piglets. The potential role of probiotic and zinc in the stimulation of the immune system is well established. Hence, the present study was undertaken to evaluate alterations of T and B cells in the small intestine after dietary inclusion of probiotic and zinc in pre and post-weaned piglets. MATERIALS AND METHODS: A total of 18 healthy Large White Yorkshire (LWY) piglets, irrespective of sex obtained from 3 litters at the age-group of 20, 30 and 60 days. They were divided into a control group fed with basal diet and a treatment group fed with probiotic and zinc supplement along with the basal diet, consisting of three animals in each group. The piglets were weaned at 28 days of age. After sacrificing the animals at day 20, 30 and 60 from both the groups, the abdominal cavity was opened and small intestinal tissue samples were collected, processed and stained by indirect immunofluorescence technique. The slides were evaluated under the fluorescent light microscope. The data were statistically analysed. RESULTS: The different T and B cell subsets were recorded in the lining epithelium, core of villus, crypt area of lamina propria and Peyer's patch area. The number of CD4+, CD8+, IgA+ and IgM+ cells was higher in the treated piglets than the control group of animals, irrespective of segments of intestine and age-group. CONCLUSIONS: It can be concluded that the dietary supplementation of probiotic and zinc was found to be good additives as they can stimulate the immune response in piglets, especially during the critical early post-weaning period.

5'-UTR-based phylogenetic analysis of Classical swine fever virus isolates from India.

Classical swine fever (CSF) caused by Classical swine fever virus (CSFV) is a globally significant disease of pigs. Genetic typing of CSFV isolates can help in understanding the epidemiology of disease and trace down the source of outbreak. 5'-UTR sequence analysis and subsequent genetic classification of nine CSFV field isolates from India indicated that 3 isolates belonged to genotype 2.1 and were closely related to European CSFV strains. The remaining 6 isolates belonged to genotype 1 that contained old and new strains. However, the genotype 2.1 group consisted of recent field isolates only. The study showed circulation of both genotypes 1 and 2.1 in north-eastern part of India.

Endotheliotropic herpesvirus infection in Asian elephants (Elephas maximus) of Assam, India.

BACKGROUND AND AIM: Elephant endotheliotropic herpesvirus (EEHV) is an emerging disease of elephant. Therefore, a study was conducted to know the actual status of the disease in Assam State of India. MATERIALS AND METHODS: A total of 289 Asian elephants of Assam were screened during 2 years of study from April 2017 to March 2019. The clinical symptoms of diseased as well as gross and histopathological changes of dead elephants were recorded for the diagnosis of the disease. Virus involved in the occurrence of the disease was confirmed by polymerase chain reaction (PCR). RESULTS: In the present study, a total of three elephant calves out of 22 were found positive to EEHV1A. On the other hand, three adult asymptomatic elephants were also found positive for EEHV1 on screening 267 captive Asian elephants of Assam. The amplified PCR product showed band size of 520, 600, and 930 bp. The PCR amplified product with size 600 bp had shown the gene sequence for EEHV1U77/HEL. Gross lesions include congested blood vessels of the liver and intestinal mucosa, foci of petechiae in the spleen, and heart and focal ulceration in the dorsal surface of the tongue. Microscopically, the kidneys showed intertubular edema and focal areas of degeneration associated with coagulative necrosis of the tubular epithelium. The liver showed hydropic degeneration and fatty changes of the hepatocytes. There was a massive proliferation of fibroblasts in the interlobular spaces which penetrated the necrosed areas of the hepatic lobules. CONCLUSION: A total of three wild rescued elephant calves and three asymptomatic adults were found positive for EEHV1A during the 2 years of study. The PCR amplified product with size 600 bp had shown the gene sequence for EEHV1U77/HEL.

Molecular characterization of shiga like toxin-producing Escherichia coli (STEC) isolates from pigs oedema.

BACKGROUND & OBJECTIVE: An oedema outbreak occurred in a Guwahati pig farm. Escherichia coli isolates from different necropsy samples collected from the dead piglets with oedema were characterized to confirm the virulence. METHODS: Haemolytic E. coli isolates recovered from liver, lung and intestine of pigs with oedema were examined for presence of genes encoding pathogroups such as enteropathogenic Escherichia coli (EPEC), (eae/bfpA), enteroaggregative Escherichia coli (EAggEC), (eagg), enterotoxigive Escherichia coil (ETEC), (elt/est) and shiga like toxin producing Escherichia coli (STEC), (stx1/ stx2) by PCR and molecular typing by randomly amplified polymorphic DNA-PCR (RAPD-PCR). RESULTS: The three haemolytic E. coli recovered from diseased pigs were STEC because of presence of the stx2 and eae genes. Analysis by RAPD-PCR indicated that two of the three isolates were genetically related. INTERPRETATION & CONCLUSION: The isolation of STEC isolates from pigs with oedema was shown. Although the three isolates were untypable, presence of eae and stx2 genes clearly indicated these as prime cause of pig oedema disease. Further, demonstration of STEC in pigs becomes a public health concern, as pigs are potential reservoir of such agents, which may cause human illness.

Classical Swine Fever in Wild Hog: Report of its Prevalence in Northeast India.

Classical swine fever virus (CSFV) is the causative agent of a highly contagious disease, hog cholera in pigs. The disease is endemic in many parts of the world and vaccination is the only way to protect the animals from CSFV infection. Wild hogs belong to the species Sus Scrofa Cristatus under the family Suidae are quite susceptible to CSFV infection. The epidemiological role concerning classical swine fever (CSF) in India is largely unknown. We report here the three isolated cases of CSF in wild hogs from three National parks, namely Kaziranga National Park, Manas National Park and Jaldapara National Park, from north-east part of India. The post-mortem and histopathological findings were clearly indicative for CSFV infection. The presence of CSFV genome was demonstrated in several organs and tissues collected from hogs died due to viral infection. In addition, CSF-specific antibodies were detected in two wild hogs as well as in eighteen feral pigs from the same locations. The phylogenetic analysis of the partial E2 protein gene and 5' untranslated region of CSFV isolates from the wild hog showed identities with genotype 2.2 of the Indian isolates. Occurrence of CSF in wild hogs may pose a potent threat in the epidemiology of the virus in Northeast part of India. To the best of our knowledge, the report presented in the manuscript is the first comprehensive report on CSF in wild hogs form Northeast India. The findings reported would help us to understand the epidemiology and biology of CSFV in wild animals.

Jejunal and ileal Peyer's patches in pigs differ in their postnatal development.

The postnatal development of the jejunal and ileal Peyer's patches was studied before and after weaning in 1-, 1.5- and 2-month-old pigs. The follicles of the jejunal Peyer's patches grew with age and were two times longer and wider in specified pathogen-free and conventional pigs than in germ-free animals, thus indicating an influence of the living microbial antigens from the gut lumen. In germ-free pigs the size of the ileal Peyer's patch follicles increased between the 1st and 2nd month, whereas in the specified pathogen-free and conventional animals these follicles were comparable in size in all three age groups. In 1- to 1.5-month-old pigs the interfollicular area of jejunal Peyer's patches was wider (0.1 +/- 0.04 mm) than that of the ileal Peyer's patch (0.04 +/- 0.03 mm). Immunohistological studies showed that in germ-free pigs preferentially surface IgM+ but few IgA+ B cells were present in the follicles, domes and dome epithelia. In specified pathogen-free and conventional pigs the B cells expressed different levels of surface or cytoplasmic IgM or IgA. In all groups studied, more T cells were observed in the jejunal than in the ileal Peyer's patch. Here, few T lymphocytes were found because of the small interfollicular areas. Small numbers of Null cells were distributed in the interfollicular regions of all animals. The results show that living microbial antigens have a major influence on the jejunal and ileal Peyer's patches in pigs. The morphological differences between the two types of Peyer's patches are an indication that they develop differently during postnatal life. So far it remains unclear whether these morphological differences reflect a specific function of the pig's ileal Peyer's patch, such as the expansion of the genetically determined B cell repertoire as has been reported for sheep.

Passive immunization of piglets against enterotoxigenic colibacillosis by vaccinating dams with K88ac pili bearing bacterins.

Day-old-piglets were passively immunized by vaccinating the pregnant sows with K88ac enterotoxigenic Escherichia coli (ETEC) vaccine. High level of ETEC specific antibodies was excreted in colostrum (3733.33 +/- 1152.13) and maintained a detectable level (100.00 +/- 0.00) up to 21 day post partum (DPP). The IgG was the predominant immunoglobulin followed by IgA and IgM. Piglets born of vaccinated dam (group A) and unvaccinated dam (group B) were challenged in 7 day of age. Clinical and faecal scores were significantly (P < 0.01) low in group A than that of group B. Piglets of group A developed mild diarrhoea (33.33%), while all the control piglets developed profuse diarrhoea and 3 of these died before 14 day of challenge infection.

Comparative efficacy of fluorescent antibody test, immunoperoxidase test and enzyme linked immuno sorbent assay in detection of rotavirus in cell culture.

Rotavirus is prevalent worldwide and has been established as a leading cause of mortality due to severe diarrhoea in neonates. Isolation of the virus is a gold standard method for confirmation of rotavirus infection in the host. Propagation of rotavirus in cell culture is a challenge as in many instances the virus does not produce detectable cytopathic effect. This study was undertaken to evaluate the efficacy of fluorescent antibody test (FAT), immunoperoxidase test (IPT) and sandwich ELISA (S-ELISA) to detect rotavirus antigen propagated in MA 104 cell line. The intensity of fluorescence and colour development for I-FAT and I-IPT was categorized and the ELISA OD values were analyzed. The overall mean of detection were 5.16 ± 0.47, 5.16 ± 0.54 and 5.66 ± 0.33 for I-FAT, I-IPT and S-ELISA, respectively. Significantly less number of samples were positive in the initial one or two passage, which increased up to 100 % from third/fourth passage onwards. The study concluded that I-FAT, I-IPT and S-ELISA were equally effective in detecting propagated rotavirus in cell line, and the former two tests are suitable for in situ demonstration of the virus while the later could be used to assay antigen in cell culture fluid.

Erythrina variegata Linn: A review on morphology, phytochemistry, and pharmacological aspects.

This review gives an account of the current knowledge on the morphology, phytochemistry, and pharmacological aspects of Erythrina variegata. E. variegata also called Erythrina indica is a thorny deciduous tree growing to 60 feet tall. A wide range of chemical compounds have been isolated, mainly alkaloids, flavonoids, triterpenoids, and lectin. Different parts of the plant have been used in traditional medicine as nervine sedative, collyrium in opthalmia, antiasthmatic, antiepileptic, antiseptic, and as an astringent. The alkaloids extracted from the leaves of E. variegata are reported to have anti-inflammatory and analgesic activity. Isoflavonoids isolated from E. variegata having antibacterial and anthelmintic activity. E. variegata shows several other characteristic pharmacological effects like neuromuscular blocking, smooth muscle relaxant, CNS depressant, and hydrocholeretic, which are consistent with the reported uses of the plant extracts in the indigenous system of medicine. Hence the present article includes the detailed exploration of morphology, phytochemistry, and pharmacological aspects of E. variegata in an attempt to provide a direction for further research.

Molecular Characterization of Classical swine fever virus Involved in the Outbreak in Mizoram.

Classical swine fever is the most insidious and devastating disease of pigs and wild boars. The virus is closely related to the other members of the genus Pestivirus. The outbreak recorded in Mizoram, India was strategically important as the state shares porous international boundary with East Asian countries. Both immunodiagnostic and molecular techniques were used to confirm the involvement of Classical swine fever virus (CSFV) in this outbreak. Sandwich ELISA and direct FAT could detect CSFV in the tissue samples. RT-nPCR specifically amplified E2 and 5'NTR product of 271 bp. Phylogenetic analysis showed, that the Mizoram isolate (MZ4/69) was very close to the Chinese strain Shimen-HVRI (93.0%) rather than other Indian isolate (CSF-30-03). Present study provides a valuable sequence based molecular data on Indian isolate of CSFV and will be useful in investigation on transmission of such disease from neighbour countries.

Characterization and expression of e2 glycoprotein of classical Swine Fever virus in a eukaryotic expression system.

Classical swine fever (CSF) is an economically important Office International des Epizooties (OIE) list A disease of swine characterized by high fever and multiple haemmorhages. The E2 glycoprotein of CSFV is immunogenic and induces neutralizing antibodies against CSFV. In the present study, complete coding region of the E2 gene from Indian virulent field isolate (Mathura) was amplified by reverse transcription-polymerase chain reaction (RT-PCR) and subsequently cloned into a mammalian expression vector; pcDNA3.1(+) at BamHI and XbaI site. The recombinant plasmid; pcDNA.E2.CSFV. was confirmed by restriction enzyme digestion. The pcDNA.E2.CSFV. transfected Vero cell expressed E2 protein which was confirmed by western blotting, immunoperoxidase and indirect immunofluorescent tests. Additionally, flow cytometry analysis also confirmed that 15% of transfected Vero cells expressed the E2 glycoprotein compared to mock or vector alone transfected cells. Further study is under way to evaluate recombinant pcDNA.E2.CSFV. Mathura clone as DNA vaccine against CSFV.