[Show abstract][Hide abstract] ABSTRACT: Kyasanur Forest disease (KFD), a tick-borne viral hemorrhagic fever, is endemic in five districts of Karnataka state, India. Recent reports of the spread of disease to neighboring districts of the Western Ghats, namely Chamarajanagar district in Karnataka, Nilgiri district in Tamil Nadu, Wayanad and Malappuram districts in Kerala, and Pali village in Goa are a cause for concern. Besides vaccination of the affected population, establishing an event-based surveillance system for monkey deaths in the national parks, wildlife sanctuaries and reserve forests of the Western Ghats would help detect the disease early and thereby help implement appropriate control measures.
[Show abstract][Hide abstract] ABSTRACT: Kyasanur Forest disease (KFD) is a febrile illness characterized by hemorrhages, and is reported endemic in the Shimoga district in Karnataka state, India. It is caused by the KFD virus (KFDV) of the family Flaviviridae, and is transmitted to monkeys and humans by Haemaphysalis ticks.
We investigated a new focus of KFD among tribals in a reserve forest in Kerala state, India. A suspected case was defined as a person presenting with acute fever, headache, or myalgia. Human sera were collected and tested for KFDV RNA by real-time RT-PCR, RT-nPCR assay, and anti-KFDV IgM and IgG by ELISA. The index case was a tribal woman with febrile illness, severe myalgia, gum bleeding, and hematemesis. Anti-KFDV IgM antibody was detected in acute and convalescent sera of the index case along with IgG in the second serum. None of her family members reported fever. On verbal autopsy, two more fatal cases were identified as probable primary cases. Acute serum from a case in the second cluster was detected positive for KFDV RNA by real time RT-PCR (Ct = 32) and RT-nPCR. Sequences of E gene showed highest similarity of 98.0% with the KFDV W-377 isolate nucleotide and 100% identity with amino acid. Anti-KFDV IgM was detected in the serum of one family member of the index case, as well as in one out of 17 other tribals.
We confirmed a new focus of KFDV activity among tribals in a reserve forest in the Malappuram district of Kerala, India.
[Show abstract][Hide abstract] ABSTRACT: Viral hemorrhagic fevers (VHFs) are major public health problems in the South-East Asia Regional (SEAR) countries. VHFs are a group of illnesses; that are caused by four families of viruses, viz. Arenaviridae, Bunyaviridae, Filoviridae and Flaviviridae. All VHFs have common features: they affect several organs and damage the blood vessels. These symptoms are often accompanied by hemorrhage. To understand pathogenesis, genetic and environmental influence that increase the risk of VHFs, efficacy and safety studies on candidate vaccines and testing of various therapeutic agents, appropriate animal models are essential tools in public and animals health. In the current review, the suitable animal models for Flavivirus [Dengue hemorhagic fever (DHF), Kyasanur forest disease (KFD)]; Bunyavirus [Crimean-Congo hemorrhagic fever (CCHF), Hantavirus fever (HF)]; and Paramyxovirus [Nipah virus fever (NiV)] have been reviewed with specific emphasis on emerging and reemerging viruses in SEAR countries.
[Show abstract][Hide abstract] ABSTRACT: Abstract Crimean-Congo hemorrhagic fever (CCHF) is a tick-borne viral disease that causes a fatal hemorrhagic illness in humans. This disease is asymptomatic in animals. CCHF was first confirmed in a nosocomial outbreak in 2011 in Gujarat State. Another notifiable outbreak occurred in July, 2013, in Karyana Village, Amreli district, Gujarat State. Anti-CCHF virus (CCHFV) immunoglobulin G (IgG) antibodies were detected in domestic animals from the adjoining villages of the affected area, indicating a considerable amount of positivity against domestic animals. The present serosurvey was carried out to determine the prevalence of CCHFV among bovine, sheep, and goat populations from 15 districts of Gujarat State, India. A total of 1226 serum samples from domestic animals were screened for IgG antibodies using a CCHF animal IgG enzyme-linked immunosorbent assay (ELISA) kit from the Centers for Disease Control and Prevention. Antibodies were detected in all the 15 districts surveyed; with positivity of 12.09%, 41.21%, and 33.62% in bovine, sheep, and goat respectively. This necessitates the surveillance of CCHFV IgG antibodies in animals and hemorrhagic fever cases in human.
[Show abstract][Hide abstract] ABSTRACT: Since the enactment of Environmental Protection Act in 1989 and Department of Biotechnology (DBT) guidelines to deal with genetically modified organisms, India has embarked on establishing various levels of biosafety laboratories to deal with highly infectious and pathogenic organisms. Occurrence of outbreaks due to rapidly spreading respiratory and haemorrhagic fever causing viruses has caused an urgency to create a safe laboratory environment. This has thus become a mandate, not only to protect laboratory workers, but also to protect the environment and community. In India, technology and science are progressing rapidly. Several BSL-3 [=high containment] laboratories are in the planning or execution phase, to tackle biosafety issues involved in handling highly infectious disease agents required for basic research and diagnosis. In most of the developing countries, the awareness about biocontainment has increased but planning, designing, constructing and operating BSL-3 laboratories need regular updates about the design and construction of facilities and clear definition of risk groups and their handling which should be in harmony with the latest international practices.
This article describes the major steps involved in the process of construction of a BSL-3 laboratory in Indian settings, from freezing the concept of proposal to operationalization phase. The key to success of this kind of project is strong institutional commitment to biosafety norms, adequate fund availability, careful planning and designing, hiring good construction agency, monitoring by experienced consultancy agency and involvement of scientific and engineering personnel with biocontainment experience in the process.
The Indian Journal of Medical Research 08/2014; 140(2):171-83. · 1.40 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Kyasanur Forest disease virus (KFDV) was first identified in 1957, when it was isolated from a sick monkey from the Kyasanur Forest in Karnataka State, India. Since then it has been reported to be enzootic in five districts of Karnataka State, India. Recent reports of human infections have reached an alarming level, in spite of the availability of a vaccine. This disease has also been reported from new areas, such as Tamil Nadu and Kerala State. During January-March 2014, KFDV-positive cases were detected in Thirthahalli taluk, Shimoga District, Karnataka State, India. Here, we report an outbreak of Kyasanur Forest disease occurring in the Kannagi [Au?2] and Konandur area, Thirthahalli taluk in Karnataka State, India, with sporadic cases from eight other areas.
International Journal of Infectious Diseases 07/2014; 26. DOI:10.1016/j.ijid.2014.05.013 · 1.86 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: During a survey in the year 2010, a novel phlebovirus was isolated from the Rousettus leschenaultii species of bats in western India. The virus was identified by electron microscopy from infected Vero E6 cells. Phylogenic analysis of the complete genome showed its close relation to severe fever with thrombocytopenia syndrome (SFTS) and Heartland viruses, which makes it imperative to further study its natural ecology and potential as a novel emerging zoonotic virus.
Journal of Virology 03/2014; 88(6). DOI:10.1128/JVI.02617-13 · 4.44 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: India is considered as a hot spot for emerging infectious diseases. In the recent past many infectious diseases of emerging and re-emerging nature have entered this subcontinent and affected a large number of populations. A few examples are Nipah, Avian influenza, Pandemic influenza, severe acute respiratory syndrome corona virus and Chikungunya virus. These diseases have not only affected human and animal health but also economy of the country on a very large scale. During December 2010, National Institute of Virology, Pune detected Crimean-Congo hemorrhagic fever virus specific IgG antibodies in livestock serum samples from Gujarat and Rajasthan states. Subsequently, during January 2011 Crimean-Congo hemorrhagic fever virus was confirmed in a nosocomial outbreak, in Ahmadabad, Gujarat, India. Retrospective investigation of suspected human samples confirmed that the virus was present in Gujarat state, earlier to this outbreak. This disease has a case fatality rate ranging from 5 to 80 %. Earlier presence of hemagglutination inhibition antibodies have been detected in animal sera from Jammu and Kashmir, the western border districts, southern regions and Maharashtra state of India. The evidences of virus activity and antibodies were observed during and after the outbreak in human beings, ticks and domestic animals (buffalo, cattle, goat and sheep) from Gujarat State of India. During the year 2012, this virus was again reported in human beings and animals. Phylogenetic analysis showed that all the four isolates of 2011, as well as the S segment from specimen of 2010 and 2012 were highly conserved and clustered together in the Asian/Middle East genotype IV. The S segment of South-Asia 2 type was closest to a Tajikistan strain TADJ/HU8966 of 1990. The present scenario in India suggests the need to look seriously into various important aspects of this zoonotic disease, which includes diagnosis, intervention, patient management, control of laboratory acquired and nosocomial infection, tick control, livestock survey and this, should be done in priority before it further spreads to other states. Being a high risk group pathogen, diagnosis is a major concern in India where only a few Biosafety level 3 laboratories exist and it needs to be addressed immediately before this disease becomes endemic in India.
Proceedings of the National Academy of Sciences, India - Section B: Biological Sciences 03/2014; 84(1). DOI:10.1007/s40011-013-0197-3 · 0.40 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Crimean-Congo hemorrhagic fever virus (CCHFV) etiology was detected in a family cluster (nine cases, including two deaths) in the village of Karyana, Amreli District, and also in a fatal case in the village of Undra, Patan District, in Gujarat State, India. Anti-CCHFV IgG antibodies were detected in domestic animals from Karyana and adjoining villages. Hyalomma ticks from households were found to be positive for CCHF viral RNA. This confirms the emergence of CCHFV in new areas and the wide spread of this disease in Gujarat State.
International journal of infectious diseases: IJID: official publication of the International Society for Infectious Diseases 11/2013; 18(1). DOI:10.1016/j.ijid.2013.09.019 · 1.86 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Background & objectives:
Pipistrellus ceylonicus bat species is widely distributed in South Asia, with additional populations recorded in China and Southeast Asia. Bats are the natural reservoir hosts for a number of emerging zoonotic diseases. Attempts to isolate bat-borne viruses in various terrestrial mammalian cell lines have sometimes been unsuccessful. The bat cell lines are useful in isolation and propagation of many of the viruses harboured by bats. New stable bat cell lines are needed to help such investigations and to assist in the study of bat immunology and virus-host interactions. In this study we made an attempt to develop a cell line from P. ceylonicus bats.
An effort was made to establish cell line from embryo of P. ceylonicus species of bat after seeding to Dulbecco's modified eagle medium (DMEM) supplemented with 10 per cent foetal bovine serum; a primary cell line was established and designated as NIV-BtEPC. Mitochondrial DNA profile analysis was done using cyt-b and ND-1 gene sequences from the cell line. Phylogenetic tree was constructed using neighbour-joining algorithm for cyt-b and ND-1 genes with 1000-bootstrap replicates.
NIV-BtEPC cell line was susceptible to Chandipura (CHPV) and novel adenovirus (BtAdv-RLM) isolated from Rousettus leschenaulti from India but did not support multiplication of a number of Bunyaviruses, Alphaviruses and Flavivirus. This might be useful for isolation of a range of viruses and investigation of unknown aetiological agents.
Interpretation & conclusions:
In this study, a new bat cell line was developed from P. ceylonicus. This cell line was successfully tested for the susceptibility to Chandipura and BtAdv-RLM virus isolated from bats. The approach developed and optimised in this study may be applicable to the other species of bats and this established cell line can be used to facilitate virus isolation and basic research into virus-host interaction.
The Indian Journal of Medical Research 08/2013; 138(2):224-31. · 1.40 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: A nosocomial outbreak of Crimean Congo hemorrhagic fever (CCHF) was reported among humans in Ahmadabad district, Gujarat, India during January, 2011. In the present study we provide the complete genomic sequences of four CCHFV isolates derived from two human patients and two pools of Hyalomma anatolicum ticks during the period of this outbreak and the complete S segment sequence of two retrospective human serum samples, positive for CCHFV in 2010. Sequence-based molecular characterization of the Indian CCHFV showed that they possessed the functional motifs known to occur in the S, M and L gene segment products as in other CCHF viruses. The S segment of the six Indian CCHF viruses showed 99.8% nucleotide identity. Notably both tick isolates shared 100% nucleotide identity with one of the Indian human isolates of 2011. Phylogenetic analysis based on the S segment demonstrated that the Indian CCHFV isolates formed a distinct cluster in the Asian-Middle East group IV of CCHF viruses. The S segment was closest to a Tajikistan strain TADJ/HU8966 of 1990 (98.5% nucleotide identity) and was of South-Asia 2 type while the M segment was of type M2. Both M and L segments were closest to an Afghanistan strain Afg09-2990 of 2009 (93% and 98% nucleotide identity respectively). The Indian isolates were thus identified as a South-Asia 2/ M2 far-east virus combination and the differing parental origin in the S and L/M segments is suggestive that it may be an intra-genotypic reassortant. Molecular clock studies further revealed that the ancestry of the viruses was not very recent and dated back to about 33 years on the basis of the S segment while it was about 15 years based on the M segment. Thus though the 2011 outbreak may not have resulted from a very recent introduction, considering that so far there is no evidence of multiple circulating strains in the country, the possibility of a recent re-introduction of the virus from any of the neighboring countries cannot be ruled out. The study thus warrants the need for continued surveillance and increased sampling of CCHFV in different parts of the country.
Infection, genetics and evolution: journal of molecular epidemiology and evolutionary genetics in infectious diseases 11/2012; 14(1). DOI:10.1016/j.meegid.2012.10.005 · 3.02 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Background & objectives:
Chittoor virus (CHITV) belongs to genus Orthobunyavirus, family Bunyaviridae. It has been isolated from various species of mosquitoes and pig from different parts of India. Five isolates of CHITV were characterized at the molecular level and compared with other Batai viruses (BATV) to find out any kind of reassortment in their genome.
Complete nucelocapsid (S), glycoprotein (M) and partial RNA polymerase (L) segments of CHITV were amplified and sequenced. These sequences were compared with those of Batai viruses, isolated from different geographical locations in Asia, Africa and Europe.
Phylogenetic analysis revealed CHITV as a variant of BATV. High level of conservation was seen among the CHITV isolates studied. The CHITV sequences showed clustering in one lineage with the sequences from Japan and Malaysia, however, BATV sequences from Europe and Africa formed a separate phylogenetic lineage.
Interpretation & conclusions:
The study indicates the presence of a single genotype of CHITV circulating in India, despite the involvement of different hosts in the natural cycle by this virus. Analysis of the sequences of the S, M and L segments of genome indicated that the virus has not undergone any reassortment. This virus has not caused any epidemic involving humans, however, replication of the virus in different mosquito and vertebrate hosts species suggests that it is a cause of concern.
The Indian Journal of Medical Research 11/2012; 136(5):792-8. · 1.40 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Kyasanur forest disease (KFD) is a zoonotic viral disease caused by infection by a Flavivirus, a member of the family Flaviviridae. KFD is a public health concern in the Karnataka State in southern India. Available conventional diagnostic tests such as virus isolation and serological tests, such as haemagglutination inhibition and complement fixation tests are time consuming. This study reports the development of a nested RT-PCR [nRT-PCR] and a TaqMan-based real-time RT-PCR and IgM antibodies capture ELISA [MAC-ELISA] for rapid and accurate diagnosis of suspected KFD cases. The nRT-PCR and the TaqMan-based real-time RT-PCR assays were developed using gene sequences of the NS-5/non-coding region. Both the assays detected KFD viral RNA in acute phase human serum samples and can provide early diagnosis of infection. Real-time RT-PCR was found to be more sensitive than nRT-PCR, which could detect 38 copies of KFDV RNA. MAC-ELISA was developed for the detection of recent infections. Although real-time RT-PCR and nRT-PCR require expensive reagents, expensive equipment and trained personnel, the developed MAC-ELISA can be used easily in the affected areas. These tests add to the existing diagnosis arsenal against haemorrhagic viruses that are prevalent in India. These assays will also help to extend our knowledge of the pathology of KFD virus and its associated clinical features, by measuring the viral titre during infection and at the time of seroconversion. Information, which is not available currently because of the lack of appropriate diagnostic methods. In addition, early laboratory diagnosis of KFDV infection will help in the application of appropriate control measures and management of KFD cases.
[Show abstract][Hide abstract] ABSTRACT: The study deals with the survey of different bat populations (Pteropus giganteus, Cynopterus sphinx, and Megaderma lyra) in India for highly pathogenic Nipah virus (NiV), Reston Ebola virus, and Marburg virus. Bats (n = 140) from two states in India (Maharashtra and West Bengal) were tested for IgG (serum samples) against these viruses and for virus RNAs. Only NiV RNA was detected in a liver homogenate of P. giganteus captured in Myanaguri, West Bengal. Partial sequence analysis of nucleocapsid, glycoprotein, fusion, and phosphoprotein genes showed similarity with the NiV sequences from earlier outbreaks in India. A serum sample of this bat was also positive by enzyme-linked immunosorbent assay for NiV-specific IgG. This is the first report on confirmation of Nipah viral RNA in Pteropus bat from India and suggests the possible role of this species in transmission of NiV in India.
The American journal of tropical medicine and hygiene 07/2012; 87(3):576-8. DOI:10.4269/ajtmh.2012.11-0416 · 2.70 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Surveillance work was initiated to study the presence of highly infectious diseases like Ebola-Reston, Marburg, Nipah and other possible viruses that are known to be found in the bat species and responsible for causing diseases in humans. A novel adenovirus was isolated from a common species of fruit bat (Rousettus leschenaultii) captured in Maharashtra State, India. Partial sequence analysis of the DNA polymerase gene shows this isolate to be a newly recognized member of the genus Mastadenovirus (family Adenoviridae), approximately 20% divergent at the nucleotide level from Japanese BatAdV, its closest known relative.
[Show abstract][Hide abstract] ABSTRACT: In January 2011, human cases with hemorrhagic manifestations in the hospital staff were reported from a tertiary care hospital in Ahmadabad, India. This paper reports a detailed epidemiological investigation of nosocomial outbreak from the affected area of Ahmadabad, Gujarat, India.
Samples from 3 suspected cases, 83 contacts, Hyalomma ticks and livestock were screened for Crimean-Congo hemorrhagic fever (CCHF) virus by qRT-PCR of which samples of two medical professionals (case C and E) and the husband of the index case (case D) were positive for CCHFV. The sensitivity and specificity of indigenous developed IgM ELISA to screen CCHFV specific antibodies in human serum was 75.0% and 97.5% respectively as compared to commercial kit. About 17.0% domestic animals from Kolat, Ahmadabad were positive for IgG antibodies while only two cattle and a goat showed positivity by qRT-PCR. Surprisingly, 43.0% domestic animals (Buffalo, cattle, sheep and goat) showed IgG antibodies in the adjoining village Jivanpara but only one of the buffalo was positive for CCHFV. The Hyalomma anatolicum anatolicum ticks were positive in PCR and virus isolation. CCHFV was isolated from the blood sample of case C, E in Vero E-6 cells and Swiss albino mice. In partial nucleocapsid gene phylogeny from CCHFV positive human samples of the years 2010 and 2011, livestock and ticks showed this virus was similar to Tajikistan (strain TAJ/H08966), which belongs in the Asian/middle east genetic lineage IV.
The likely source of CCHFV was identified as virus infected Hyalomma ticks and livestock at the rural village residence of the primary case (case A). In addition, retrospective sample analysis revealed the existence of CCHFV in Gujarat and Rajasthan states before this outbreak. An indigenous developed IgM ELISA kit will be of great use for screening this virus in India.