Investigation of buffalopox outbreaks in Maharashtra State during 1992-1996.

During 1992-96, outbreaks of buffalopox zoonosis were reported from different villages in Jalgaon, Dhule and Beed districts of Maharashtra State. In humans, pox lesions were observed on the hands whereas in affected buffaloes and cows the lesions were noticed mainly on the teats and udder. Twenty two virus strains were isolated from the skin scabs collected from infected humans and milch animals. Neutralizing antibodies were detected not only in the sera of affected humans but also in their contacts. Detection of antibodies in young individuals from endemic area, who were neither vaccinated for smallpox nor had any contact with buffaloes or history of any poxvirus disease, is suggestive of occurrence of subclinical infection. A few children who had no contact with infected animals also showed clinical manifestations with disseminated lesions on the face, arm and buttocks, and thus suspected to have acquired infection through their infected parents or other family members indicating a possible man to man transmission. Therefore, in the light of discontinuation of smallpox vaccination, buffalopox outbreaks need to be monitored carefully as this may emerge as a serious zoonotic disease in India.

Susceptibility of laboratory-bred rodents to the experimental infection with dengue virus type 2.

Various strains of laboratory-bred rodents viz. mice [Swiss, C57BL/6, C3H/Hej, DBA/2, BALB/c, NMRI (nu/nu) and BL6 (nu/nu) and their heterozygous siblings (nu/+)], Mastomys natalensis, Wistar rat, golden hamster and Indian desert gerbil were inoculated intracerebrally (ic) with mouse-adapted dengue virus type 2 (DV-2). The inoculated animals were observed daily for dullness, anorexia, occult blood in faeces, patechial haemorrhages, lacrymation, paralysis, cachexia, death. Necropsied animals were examined for gastrointestinal haemorrhages and lymphadenopathy. The severity of clinical symptoms in various rodents declined as follows: (i) BL6 (nu/nu) mice exhibited most severe manifestation of all the aforementioned symptoms followed by (ii) NMRI (nu/nu), (iii) BL6 (nu/+) (iv) NMRI (nu/+) and C57BL6, (v) DBA, C3H/Hej and BALB/c, and (vi) Swiss. These results indicate that adaptation of DV-2 to the mouse may be an important factor in exaltation of virulence. Interstrain variation in manifestation of symptoms in mice indicates that the susceptibility to DV-2 may be determined by host genetic factors.

Induction of tumour necrosis factor alpha in experimental animals treated with hepatotoxicants.

Administrations of hepatotoxicants namely carbon tetrachloride (CCl4:0.4 ml in 1.2 ml of liquid paraffin) and ANIT (1 ml of 1.5% solution in liquid paraffin) in Charles foster rats (force fed) and D-galactosamine (8 mg in water per swiss albino mouse, ip) induce the release of TNF-alpha in case of CCl4 and D-galactosamine. High TNF-alpha level was observed up to 48 hr in CCl4 and up to 24 hr in D-galactosamine treated animals. Elevated levels of biochemical like ALP and SGPT are also recorded. TNF-alpha level can be measured of tissue damage and prognosis in case of hepatitis.

Down regulation of tumour necrosis factor activity in experimental hepatitis by a herbal formulation, Liv. 52.

A herbal hepatoprotective formulation Liv 52 down regulated the tumour necrosis factor (TNF) production in Charles Foster Rats treated with CCl4. Inhibition of TNF activity was proportional to the hepatoprotective activity.

Antiviral activity of the Indian medicinal plant extract Swertia chirata against herpes simplex viruses: a study by in-vitro and molecular approach.

PURPOSE: The antiviral activity of Indian Medicinal plant extract Swertia chirata was tested against Herpes simplex virus (HSV) type-1, using multiple approaches both at cellular and molecular level. METHODS: Cytotoxicity, plaque reduction, virus infectivity, antigen expression and polymerase chain reaction (PCR) assays were conducted to test the antiviral activity of the plant extract. RESULTS: Swertia plant crude extract (1 gm/mL) at 1:64 dilution inhibited HSV-1, plaque formation at more than 70% level. HSV antigen expression and time kinetics experiments conducted by indirect immunofluorescence (IFA) test, revealed a characteristic pattern of small foci of single fluorescent cells in Swertia extract treated HSV-1 infected cells at 4 hours post infection dose, suggested drug inhibited viral dissemination. Infected cell cultures treated with Swertia extract at various time intervals, tested by PCR, failed to show amplification at 12, 24-72 hours. HSV-1 infected cells treated with Acyclovir (antiviral drug) did not show any amplification by PCR. CONCLUSIONS: In this preliminary study, the Indian medicinal plant extract, Swertia chirata showed antiviral properties against Herpes simplex virus type-1.

Outbreak of acute hemorrhagic conjunctivitis in Maharashtra and Gujarat states of India, caused by Coxsackie virus A-24 variant.

Acute hemorrhagic conjunctivitis is associated with enteroviruses. Among these, Coxsackie A-24 variant (CA-24) and Enterovirus-70 (EV-70) are known to cause epidemics and pandemics. An outbreak of acute hemorrhagic conjunctivitis occurred in August-September 2003 in Maharashtra and Gujarat states of India. The present investigation was carried out to determine the viral etiological agent associated with the epidemic. Virus isolates were obtained from 11 eye swabs of conjunctivitis patients using HeLa/ Hep-2 cell lines. The isolates were characterized by serological and mouse pathogenecity tests, RT-PCR using enterovirus common primers (VP4-VP2), CA-24 specific primers (3C-proteinase region), EV-70 primers (VP-3) followed by sequencing, and phylogenetic analysis. The virus was characterized as a Coxsackie A-24 variant (CA-24v) and none of the isolates were found to be positive for EV-70. Sequencing of the PCR products derived from all the 11 isolates revealed 98.4% (SE 0.20) nucleotide identity within the Indian strains and 98.6% (0.50) and 94.4% (0.30) nucleotide identity respectively with the West Indies and Asian strains reported worldwide. The findings suggest that the outbreak of acute hemorrhagic conjunctivitis that occurred in Maharashtra and Gujarat states of India during August-September 2003 was caused by the Coxsackie A-24 variant (CA-24v).

Xenogenic transfer of human lymphocytes in tolerized mice.

Female nu/+ or BALB/c mice were immunized with human peripheral blood lymphocytes (PBL) before and during pregnancy. Pups born to these mothers were inoculated with human PBL or human fetal bone marrow and thymus cells. Tolerization of the pups to human PBL was observed without graft-versus-host reaction. Presence of human immunoglobulins was observed in the pups for 3-4 weeks. Human T cells also could be detected for a period of 3-4 months in these mice.