Browsing by Author "Mishra, A C"
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Item Antigen distribution pattern of Japanese encephalitis virus in Culex tritaeniorhynchus, C. vishnui & C. pseudovishnui.(2000-05-16) Mourya, D T; Mishra, A CDistribution of Japanese encephalitis (JE) virus antigen in different tissues of mosquitoes was studied in three species viz., Culex tritaeniorhynchus, C. Vishnui and C. pseudovishnui. Overall per cent positivity was higher in the intrathoracically infected as compared to the orally infected mosquitoes, suggesting the existence of a midgut barrier. The cells at the junction of the foregut-midgut, and midgut-hindgut showed intense fluorescence from the second day post feeding onwards. This suggests that the dissemination of virus takes place from these regions of the gut. A small number of salivary glands were found negative even though fluorescence was seen in the respective head squashes, suggesting involvement of the salivary gland barrier in these mosquitoes. Though there was no difference in the salivary gland positivity between these three species, the salivary gland area positivity was high in C. tritaeniorhynchus mosquitoes. Presence of virus antigen in the ovaries and developing eggs of these three species on the third day suggests the possibility of transovarial transmission of virus even in the first gonotrophic cycle which may have epidemiological importance.Item Antigen distribution pattern of West Nile virus in Culex tritaeniorhynchus, Culex vishnui and Culex pseudovishnui mosquitoes.(2001-09-11) Mishra, A C; Jadi, R S; Paramasivan, R; Mourya, D TDistribution of West Nile (WN) virus antigen in different tissues of mosquitoes was studied in three species viz., Culex tritaeniorhynchus, C. vishnui and C. pseudovishnui. Overall per cent positivity was higher in the intra thoracically inoculated as compared to the orally infected mosquitoes, suggesting the existence of a midgut barrier. In a small number of mosquitoes salivary glands were found negative even though fluorescence was seen in the respective head squashes, suggesting salivary gland barrier in these mosquitoes. There was no difference in the per cent salivary gland and salivary gland area positivity between these three species. Presence of virus antigen in the ovaries of these three species on the 3rd post infection day suggests the possibility of transovarial transmission of virus even in the first gonotrophic cycle, which is of epidemiological importance.Item Antiviral drug profile of human influenza A & B viruses circulating in India: 2004-2011.(2014-08) Potdar, V A; Dakhave, M R; Kulkarni, P B; Tikhe, S A; Broor, S; Gunashekaran, P; Chawla-Sarkar, M; Abraham, A; Bishwas, D; Patil, K N; Kadam, A A; Kode, S S; Mishra, A C; Chadha, M Sbackground & objectives: Recent influenza antiviral resistance studies in South East Asia, Europe and the United States reveal adamantane and neuraminidase inhibitor (NAIs) resistance. This study was undertaken to evaluate antiviral resistance in influenza viruses isolated from various parts of India, during 2004 to 2011. methods: Influenza viruses were analyzed genetically for known resistance markers by M2 and NA gene sequencing. Influenza A/H1N1 (n=206), A/H3N2 (n=371) viruses for amantadine resistance and A/H1N1 (n=206), A/H3N2 (n=272) and type B (n=326) for oseltamivir resistance were sequenced. Pandemic (H1N1) (n= 493) isolates were tested for H274Y mutation by real time reverse transcription (rRT)-PCR. Randomly selected resistant and sensitive influenza A/H1N1 and A/H3N2 viruses were confirmed by phenotypic assay. results: Serine to asparagine (S3IN) mutation was detected in six isolates of 2007-2008.One dual-resistant A/H1N1 was detected for the first time in India with leucine to phenylalanine (L26F) mutation in Mm2 gene and H274Y mutation in NA gene. A/H3N2 viruses showed an increase in resistance to amantadine from 22.5 per cent in 2005 to 100 per cent in 2008 onwards with S3IN mutation. Fifty of the 61 (82%) A/H1N1 viruses tested in 2008-2009 were oseltamivir resistant with H274Y mutation, while all A/H3N2, pandemic A/H1N1 and type B isolates remained sensitive. Genetic results were also confirmed by phenotypic analysis of randomly selected 50 resistant A/H1N1 and 40 sensitive A/H3N2 isolates. Interpretation & conclusions: Emergence of influenza viruses resistant to amantadine and oseltamivir in spite of negligible usage of antivirals emphasizes the need for continuous monitoring of antiviral resistance.Item Beta hemolytic streptococci in sore throat infections.(1997-06-01) Srivastava, S P; Kumar, A; Mishra, A C; Bharti, L KItem Biochemical basis of DDT-resistance in Aedes aegypti population from a dengue affected area in Shahjahanpur city.(1994-05-01) Mourya, D T; Gokhale, M D; Mishra, A CEntomological studies conducted in Jalalnagar, Shahjahanpur city, Uttar Pradesh, India, during an outbreak of dengue in 1992, showed that Ae. aegypti mosquitoes were resistant to DDT and had some tolerance to malathion in the adults and the larvae. Biochemical analysis suggested that DDT resistance was related to elevated glutathione s-transferase and tolerance to malathion was due to a little increase in esterase activity. Crosses of DDT-resistant and susceptible strain suggested that resistance was codominant (metabolic type).Item Chandipura virus growth kinetics in vertebrate cell lines, insect cell lines & embryonated eggs.(2010-08) Jadi, R S; Sudeep, A B; Kumar, Satyendra; Arankalle, V A; Mishra, A CBackground & objectives: Since not much information on Chandipura virus is available, an attempt was made to study the growth kinetics of the virus in certain vertebrate, invertebrate cell lines and embryonated chicken eggs. Methods: Comparative study of Chandipura virus (CHPV) growth kinetics in three vertebrate cell lines [Vero E6, Rhabdo myosarcoma (RD), Porcine stable kidney (PS) cell lines], two insect cell lines [Aedes aegypti (AA) and Phlebotomus papatasi (PP-9) cell lines] and embryonated pathogen free chicken eggs was conducted, by tissue culture infective dose 50 per cent (TCID50) and indirect immunofluorescence assay (IFA). Results: All the cell lines and embryonated egg supported the growth of CHPV and yielded high virus titre. The vertebrate cell lines showed distinct cytopathic effect (CPE) within 4-6 h post infection (PI), while no CPE was observed in insect cell lines. PP-9 cell line was the most sensitive system to CHPV as viral antigen could be detected at 1 h PI by IFA. Interpretation & conclusions: Our results demonstrated that all the systems were susceptible to CHPV and achieved high yield of virus. However, the PP-9 cell line had an edge over the others due to its high sensitivity to the virus which might be useful for detection and isolation of the virus during epidemics.Item Characterization of a newly established potato tuber moth (Phthorimaea operculella Zeller) cell line.(2005-03-02) Sudeep, A B; Khushiramani, R; Athawale, S S; Mishra, A C; Mourya, D TBACKGROUND & OBJECTIVE: Potato tuber moth (PTM), Phthorimaea operculella Zeller is a widely distributed, devastating pest of potatoes attacking the foliage and infest the tubers in both field and store causing serious economic damage. As application of PTM granulovirus (PTM-GV) has shown significant reduction in damage, attempts were made to develop a new cell line from this insect to grow PTM-GV for use as a biopesticide. METHODS: Approximately 100 mg of insect eggs were collected, surface sterilized and crushed gently in a boiling tube aseptically. The tissues were washed with physiological saline, suspended in growth medium and incubated stationary at 28 degrees C. Morphology of cells was studied after staining with Giemsa. Besides karyological and growth curve studies, PCR amplification was also done for rapid amplified polymorphic DNA pattern. RESULTS: A new cell line from the embryonic tissue of PTM was maintained in Mitsuhashi Maramorosch medium supplemented with 10 per cent foetal bovine serum. It is in the 78th passage level and designated as NIV-PTM-1095. Random amplified polymorphic DNA profile analysis indicated this as a new cell line from potato tuber moth and differed from the profiles of two other lepidopteran cell lines maintained in the laboratory. Three different cell types were observed at the 40th passage level and comprised of epithelial-like cells (77%), fibroblast-like cells (20%) and giant cells (3%). The chromosome number varied from 54-176. The cell line had a cell doubling time of approximately 42 h during the logarithmic phase of growth. The cell line did not support the multiplication of any of the baculoviruses used in the study. INTERPRETATION & CONCLUSION: Since the new cell line is found to replicate PTM-GV, it may be useful for the propagation of PTM-GV in large scale. Studies to scale up the production of the GV in the cell line and field trials may lead to its widespread use as an eco-friendly biopesticide.Item Double blind clinical trial on centperazine & DEC in bancroftian filariasis.(1990-07-01) Rath, R N; Das, B K; Ali, M L; Das, P K; Mishra, A C; Mohapatra, B NCentperazine, an analogue of DEC, was subjected to a double blind controlled trial, to evaluate its efficacy as a newer antifilarial agent. Centperazine (300 mg/day) along with equivalent quantities of DEC and placebo were administered to different types of filariasis patients. DEC was found to be significantly effective in reducing peripheral microfilaraemia, in different weeks and months of follow-up, except at the end of 6th month, as compared to Centperazine. There was no significant difference between the placebo and Centperazine treated patients, in this respect, revealing that the drugs had no efficacy in eliminating peripheral microfilaraemia. Recurrence of acute attack within 6 months was nearly equal with both Centperazine and DEC, being 28.2 and 24 per cent respectively, whereas in the placebo group the recurrence rate was 48.9 per cent. Centperazine treated patients showed significantly less side effects (8.9%), as compared to DEC treated patients (34%). Giddiness, nausea and vomiting were the common adverse effects observed.Item Effect of temperature and insecticide stresses on Aedes aegypti larvae and their influence on the susceptibility of mosquitoes to dengue-2 virus.(2005-09-28) Yadav, P; Barde, P V; Gokhale, M D; Vipat, V; Mishra, A C; Pal, J K; Mourya, D TTwo major factors, higher temperatures and the application of insecticides, can drastically alter the genetic structure of a vector mosquito population. Due to these two stresses, the majority of the population gets wiped out, but the ones that withstand the stress and survive are likely to pass on survivability, and have an altered physiology. Our study shows that exposures to higher temperatures and DDT during the larval stage affects their susceptibility as adult mosquitoes to the DEN-2 virus. The overall transcription and translation status of heat shock protein (Hsp70) in virus high- and low-susceptible was the same as that in other batches. In the case of a DDT-resistant (R-7) strain two bands were obtained during RT-PCRs after heat shock. These two alleles were obtained only with HY-1 in which R-7 males were used for the crosses, suggesting that the second allele is probably male sex linked. The higher expression of Hsp70 may provide DDT-resistant strains a better chance of survival high temperature environments, particularly in homozygotes and hybrids. It was also interesting to note that these strains have a significantly lower susceptibility to the virus. Wide-spread DDT-resistance and a rise in temperature above the average temperature during summer may result in a population with a low susceptibility to the virus. Several families of heat shock proteins are known to be expressed in mosquitoes, and may have a cumulative role in determining susceptibility to the virus, which itself is governed by several genes.Item Establishment of two new cell lines from Bombyx mori (L.) (Lepidoptera: Bombycidae) & their susceptibility to baculoviruses.(2002-05-05) Sudeep, A B; Mishra, A C; Shouche, Y S; Pant, U; Mourya, D TBACKGROUND & OBJECTIVES: Lepidopteran cell cultures and baculovirus expression vector systems are becoming popular due to their potential applications in biotechnology especially for the expression of foreign proteins. Efforts were made to develop new, indigenous, cell lines from Bombyx mori larvae and pupae. METHODS: Eight to ten B. mori larvae and 10-12 pupae were surface sterilized, dissected and ovaries were removed aseptically. Ovaries were chopped finely, washed and suspended in growth medium. When the cells formed monolayers, they were subcultured and experiments were carried out. RESULTS: Two new cell lines from larval and pupal ovaries of B. mori were established in Grace's insect tissue culture medium supplemented with 20 per cent FBS (foetal bovine serum). The larval cell line consisted predominantly of epithelial-like cells (98.31%), whereas the pupal cell line had a mixed cell population of epithelial-like (71.8%) and fibroblast-like cells (27.8%). Karyology indicated a typical lepidopteran pattern in both the cell lines and had chromosome numbers ranging from 35 to 150 and 60 to 180 for larval and pupal ovaries respectively. Four-fold increase in cell number was observed in these cell lines in 7 days. Both the cell lines were found susceptible to B. mori multiple nucleopolyhedrovirus and Autographa californica multiple nucleopolyhedrovirus, but not to Helicoverpa armigera single nucleopolyhedrovirus and Spodoptera litura multiple nucleopolyhedrovirus. INTERPRETATION & CONCLUSION: These well characterized cell lines may be of immense application in biotechnology and medicine for the production of biologically active recombinant proteins to use in vaccine studies as well as in therapeutic applications.Item Experimental transmission of Japanese encephalitis virus by Culex vishnui Theobald.(1984-03-01) Banerjee, K; Mishra, A C; Bhat, H RItem A focal outbreak of Japanese Encephalitis among horses in Pune district, India.(2003-03-09) Raut, C G; Thakare, J P; Padbidri, V S; Sapkal, G N; Mishra, A C; Paramasivan, R; Gokhale, M D; Mourya, D T; Shouche, Y S; Jayakumar, P CItem Ganjam virus.(2009-11) Sudeep, A B; Jadi, R S; Mishra, A CGanjam virus (GANV), a member of genus Nairovirus of family Bunyavirdae is of considerable veterinary importance in India. Though, predominantly tick borne, GANV was also isolated from mosquitoes, man and sheep. Neutralizing and complement fixing antibodies to GANV have been detected in animal and human sera collected from different parts of the country. Thirty three strains of GANV have been isolated from India, mainly from Haemaphysalis ticks. The virus replicated in certain vertebrate and mosquito cell lines and found pathogenic to laboratory animals. One natural infection and five laboratoryacquired infections in men were also reported. GANV is antigenically related to Nairobi sheep disease virus (NSDV) of Africa, which is highly pathogenic for sheep and goats causing 70-90 per cent mortality among the susceptible population. Recent molecular studies have demonstrated that GANV is an Asian variant of NSDV and both these viruses are related to the dreaded Crimean Congo haemorrhagic fever (CCHF) group viruses. The versatility of the virus to replicate in different arthropod species, its ability to infect sheep, goat and man makes it an important zoonotic agent.Item Immunopathological changes in kidney in Plasmodium falciparum malaria.(1990-03-01) Rath, R N; Patel, D K; Das, P K; Das, B K; Mishra, A C; Sahu, R N; Mohanty, GFifteen (34.8%) of 43 patients of falciparum malaria screened for urinary abnormalities showed significant proteinuria (greater than 150 mg/24 h), haematuria (greater than 1/HPF) and casts, with or without azotaemia. Light microscopic examination of renal biopsy tissue from 12 patients revealed mesangial and endothelial proliferative change in 8, and acute tubular necrosis in one patient. Immunofluorescence showed IgM alone, or IgG and IgM along with C3, in 7 patients within the mesangium or along the capillary walls. Repeat kidney biopsy after 6 wk in 5 patients revealed no residual pathology indicating the reversible nature of the lesions.Item Influenza virus genotypes circulating in and around Lucknow, Uttar Pradesh, India, during post pandemic period, August 2010 - September 2012.(2014-03) Dangi, Tanushree; Jain, Bhawana; Singh, Ajay Kumar; Mohan, Madan; Dwivedi, Mukesh; Singh, J V; Kumar, Rashmi; Singh, K P; Chaddha, M S; Mishra, A C; Jain, AmitaBackground & objectives: During the post influenza pandemic period, continuous surveillance of influenza virus and its subtypes is mandatory to help the policy makers to take effective and appropriate decisions. Therefore, this study was planned to determine the pattern of influenza virus activity in context to various meteorological and clinical parameters in and around Lucknow, Uttar Pradesh, India, during post pandemic period August 2010 - September 2012. Methods: Nasal swabs/throat swabs/nasopharyngeal aspirates of 2669 patients were collected. One-step real time PCR for detection of influenza virus was done according to the Centers for Disease Control and Prevention (CDC) protocol. Results: Influenza positivity was 15.8 per cent (423/2669) in symptomatic patients. Of the 423 total positives, 192 (7.2%) were influenza A and 231 (8.7%) were influenza B. Positivity for influenza virus was significantly (P=0.001, OR=2.9, CI=1.9-4.3) higher in patients with Influenza like illness (ILI) (17.4%, 396/2271) than those with severe acute respiratory illness (SARI) (6.8%, 27/398). Influenza A positive samples were subtyped as; pdmH1N1 (67.2%, 129/192) and seasonal H3N2 (32.8%, 63/192). It significantly correlated with monthly mean rainfall, humidity and dew point while atmospheric pressure was inversely related. No significant association was found with temperature and wind speed. Clinical variations were observed between different strains of Influenza virus. Interpretation & conclusions: The findings provide a clear picture of different clinical presentations of various strains of influenza A and B viruses and epidemiology of influenza infection from Lucknow (UP), India. The seasonality of influenza virus infection showed variation in relation to different environmental factors. Pandemic H1N1 caused more systemic infection than seasonal influenza A/H3N2 virus.Item Insect cell culture in research: Indian scenario.(2005-06-23) Sudeep, A B; Mourya, D T; Mishra, A CInsect cell cultures are widely used in viral diagnosis and biotechnology, for the production of recombinant proteins, viral pesticides and vaccines as well as in basic research in genetics, molecular biology, biochemistry, endocrinology and virology. Following KRP Singh's pioneering research in 1967, a large number of cell lines from diptera, hemiptera, and lepidopteran insects were established and characterized in India. With the availability of the modern tools in molecular biology and the advancements made in biotechnology, the indigenous cell lines may prove useful in creating a future without biohazardous chemical pesticides as well as producing life saving pharmaceuticals and vaccines for many diseases. This review summarizes information gathered regarding the insect cell lines established so far in India. It also covers the familiarization of the well characterized continuous cell lines and their potential applications. Special attention is given to virus susceptibility of the cell lines, the yield of virus with a comparative analysis with other conventional systems. The potential applications of dipteran and lepidopteran cell lines in agriculture and biotechnology are also briefly discussed for prospective studies.Item Isolation of Chandipura virus (Vesiculovirus: Rhabdoviridae) from Sergentomyia species of sandflies from Nagpur, Maharashtra, India.(2014-05) Sudeep, A B; Bondre, V P; Gurav, Y K; Gokhale, M D; Sapkal, G N; Mavale, M S; George, R P; Mishra, A CBackground & objectives: An outbreak of acute encephalitis syndrome was reported from Vidarbha region of Maharashtra state, India, during July 2012. Anti-IgM antibodies against Chandipura virus (CHPV) were detected in clinical samples. Sandfly collections were done to determine their role in CHPV transmission. Methods: Twenty nine pools of Sergentomyia spp. comprising 625 specimens were processed for virus isolation in Vero E6 cell line. Diagnostic RT-PCR targeting N-gene was carried out with the sample that showed cytopathic effects (CPE). The PCR product was sequenced, analysed and the sequences were deposited in Genbank database. Results: CPE in Vero E6 cell line infected with three pools was detected at 48 h post infection. However, virus could be isolated only from one pool. RT-PCR studies demonstrated 527 nucleotide product that confirmed the agent as CHPV. Sequence analysis of the new isolate showed difference in 10-12 nucleotides in comparison to earlier isolates. Interpretation & conclusions: This is perhaps the first isolation of CHPV from Sergentomyia spp. in India and virus isolation during transmission season suggests their probable role in CHPV transmission. Further studies need to be done to confirm the precise role of Sargentomyia spp. in CHPV transmission.Item Isolation of Japanese encephalitis virus from mosquitoes collected in Bankura district (West Bengal) during October 1974 to December 1975.(1979-02-01) Banerjee, K; Mahadev, P V; Ilkal, M A; Mishra, A C; Dhanda, V; Modi, G B; Geevarghese, G; Kaul, H N; Shetty, P S; George, P JItem Molecular characterization of Chittoor (Batai) virus isolates from India.(2012-11) Yadav, P D; Sudeep, A B; Mishra, A C; Mourya, D TBackground & 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. Methods: 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. Results: 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.Item Mosquito vectors of Japanese encephalitis epidemic (1983) in Mandya district (India).(1984-10-01) Mishra, A C; Jacob, P G; Ramanujam, S; Bhat, H R; Pavri, K M