Virus Research Centre Research Articles

Prevalence and Characteristics of Influenza Cases From 2017 to 2019 at a Tertiary Care Teaching Hospital in Karnataka.

Introduction Influenza virus is a significant human pathogen causing severe acute respiratory illness (SARI) associated with significant mortality worldwide. The H1N1 Influenza virus that caused a pandemic in 2009 continued to cause periodic epidemics worldwide, with new variants posing significant public health problems. The present study was carried out to determine the prevalence and characteristics of influenza at a tertiary care teaching hospital. Methods From 2017 to 2019, respiratory samples from suspected cases of influenza belonging to category C received at the microbiology laboratory were transported to Manipal Centre for Virus Research, Manipal,in the cold chain for testing of influenza virus by real-time reverse transcriptase polymerase chain reaction (rRT-PCR) as per CDC guidelines. The microbiological reports were collected and evaluated. The details of patients positive for influenza were analyzed for demographic and clinical characteristics. Results During the study period, 172 samples from SARI patients were tested, out of which 44 patients were positive for the influenza virus, accounting for a prevalence of 25.58%; 84% (n=37) of the cases were infected with H1N1 influenza virus, and the other 11.36% (n=5) and 4.54% (n=2) cases yielded H1N2 and H1N3 influenza virus, respectively. Among 44 patients, 56.81% (n=25) were females and 43.18% (n=19) were males. Most of the patients, 65.9% (n=29), were between 40 and 60 years old. The predominant presenting symptoms were fever in 81.81% (n=36) patients, breathlessness in 56.8% (n=25) patients, and cough in 54.54% (n=24) patients. Twelve (27.27%) patients had acute severe respiratory distress syndrome (ARDS). A significant mortality rate of 22.72% (n=10) was noted in the study. Conclusion A significant prevalence of influenza was noted in the study at 25.58%. Along with the H1N1 Influenza virus, the new strains detected in our region were H1N2 and H1N3 influenza viruses. Regular surveillance is important in the early detection of cases, for timely management, to reduce mortality, and to take measures to prevent the spread of this important infectious disease.

Microwave hyperthermia represses human papillomavirus oncoprotein activity and induces cell death due to cell stress in 3D tissue models of anogenital precancers and cancers

Hyperthermia is a well-accepted cancer therapy. Microwaves provide a very precise, targeted means of hyperthermia and are currently used to treat plantar warts caused by cutaneous-infective human papillomaviruses (HPVs). Other HPV genotypes infecting the anogenital mucosa cause genital warts or preneoplastic lesions or cervical cancer. Effective, non-ablative therapies for these morbid HPV-associated lesions are lacking. The molecular consequences of microwave treatment were investigated in invitro cultured three-dimensional HPV-positive cervical tumour tissues, and tissues formed from HPV-infected normal immortalised keratinocytes. Microwave energy delivery to tissues was quantified. Quantitative reverse transcriptase PCR was used to quantify mRNA expression. Immunohistochemistry and fluorescence immunostaining was used to assess protein expression. Microwave energy deposition induced sustained, localised cell death at the treatment site. There was a downregulation in levels of HPV oncoproteins E6 and E7 alongside a reduction in cellular growth/proliferation and induction of apoptosis/autophagy. HSP70 expression confirmed hyperthermia, concomitant with induction of translational stress. The data suggest that microwave treatment inhibits tumour cell proliferation and allows the natural apoptosis of HPV-infected cells to resume. Precision microwave delivery presents a potential new treatment for treating HPV-positive anogenital precancerous lesions and cancers. Funding was through an Innovate UK Biomedical Catalyst grant (ID# 92138-556187), a Chief Scientist Office grant (TCS/19/11) and core support from Medical Research Council (MC_ UU_12014) core funding for the MRC-University of Glasgow Centre for Virus Research.

External quality assurance of serological diagnosis of dengue, chikungunya and Japanese encephalitis virus infection

Dengue, chikungunya and Japanese encephalitis are the most common arthropod-borne viral diseases in India. Due to overlapping clinical symptoms, accurate, high-quality and timely laboratory-based differential diagnosis is essential for control and containment of outbreaks. This is most commonly done by detection of IgM antibodies in serum using enzyme-linked immunosorbent assays. The Resource Centre for Virus Research and Diagnostic Laboratories (VRDLs) in Pune, India organized an external quality assurance (EQA) study to check the accuracy of serological diagnostics in the VRDL network. Three panels, one each for anti-dengue virus, anti-chikungunya virus and anti-Japanese encephalitis virus IgM antibodies, comprising six human serum samples (two positive and four negative) were distributed to test the sensitivity, specificity and reproducibility of serological testing in 124 VRDLs across India in 2018-19 and 2019-20. Among the 124 VRDLs, the average concordance for both 2018-19 and 2019-20 was 98%. In 2018-19, 78.33%, 13.33% and 6.66% of VRDLs reported 100% concordance, 91-99% concordance and 81-90% concordance with the reference results, respectively, and 1.66% of VRDLs had concordance <80%. In 2019-20, 79.68%, 14.06% and 4.68% of VRDLs reported 100% concordance, 91-99% concordance and 81-90% concordance with the reference results, respectively, and 1.56% of VRDLs had concordance <80%. The EQA programme was beneficial for assessing and understanding the performance of the VRDLs. The study data indicate good proficiency in serological diagnosis of dengue, chikungunya and Japanese encephalitis in the VRDL network laboratories. Further expansion of the EQA programme to cover other viruses of public health importance will increase confidence among the VRDL network, and generate evidence of high-quality testing.

Clinical profile and outcome of H1N1 influenza patients in a tertiary care hospital in Kochi, Kerala

Background: After the 2009 pandemic, a re-emergence of hemagglutinin type 1 and neuraminidase type 1 (H1N1) influenza cases has been noted in India recently. The number of swab-positive cases has increased in the year 2017 compared to yesteryears. Since the current circulating strain (A/Michigan/7/2009 [H1N1] pdm09) is different from previous pandemic strains, a look into the clinical profile is imperative. Aims: The aim is to study the clinical, biochemical, and radiological profile of H1N1 patients at initial presentation and its influence on the mode of treatment and outcome. Patients and Methods: A cross-sectional record-based analysis of all confirmed cases of H1N1 influenza admitted at Lourdes Hospital, Kochi, Kerala, between 2015 and 2017. Confirmation of cases was done by reverse transcriptase polymerase chain reaction of respiratory specimens at Manipal Centre for Virus Research. Results: A total of 76 confirmed cases of H1N1 influenza were detected during the study period of which 36 required Intensive Care Unit admission. Most patients were between 51 and 60 years (25%). The predominant presenting symptoms were fever (98.7%), dry cough (61.8%), breathlessness (53.9%), and the most common auscultatory finding being bilateral crepitations (64.47%). Around 32.89% of cases presented with bilateral lung infiltrates on X-ray. Sixty-nine of 76 patients (90.79%) survived the disease. Conclusions: Vaccination, early recognition of the disease, and prompt initiation of treatment seem to be the only way to reduce H1N1 disease progression and associated mortality. Patients with risk factors require additional attention as clinical course can be unpredictable. Pregnancy is associated with higher rate of complications. Early respiratory support helped in preventing progression to respiratory failure in most of our patients.

Characterisation of a Hepatitis C Virus Subtype 2a Cluster in Scottish PWID with a Suboptimal Response to Glecaprevir/Pibrentasvir Treatment.

Direct-acting antivirals (DAAs) have revolutionised the treatment of Hepatitis C virus (HCV), allowing the World Health Organisation (WHO) to set a target of eliminating HCV by 2030. In this study we aimed to investigate glecaprevir and pibrentasvir (GP) treatment outcomes in a cohort of patients with genotype 2a infection. Methods: Clinical data and plasma samples were collected in NHS Greater Glasgow & Clyde. Next generation whole genome sequencing and replicon assays were carried out at the MRC-University of Glasgow Centre for Virus Research. Results: 132 cases infected with genotype 2a HCV were identified. The SVR rate for this group was 91% (112/123) following treatment with GP. An NS5A polymorphism, L31M, was detected in all cases of g2a infection, and L31M+R353K in individuals that failed treatment. The results showed that R353K was present in 90% of individuals in the Glasgow genotype 2a phylogenetic cluster but in less than 5% of all HCV subtype 2a published sequences. In vitro efficacy of pibrentasvir against sub-genomic replicon constructs containing these mutations showed a 2-fold increase in IC50 compared to wildtype. Conclusion: This study describes a cluster of HCV genotype 2a infection associated with a lower-than-expected SVR rate following GP treatment in association with the NS5A mutations L31M+R353K.

Towards global health security: response to the May 2018 Nipah virus outbreak linked to Pteropus bats in Kerala, India

### Summary box This report summarises the complete outbreak timeline and findings, including the association of infection and Pteropus bats, and makes the link to global health security in India....

Outbreak of Nipah Virus Disease in Kerala, India, 2018

Background: Nipah Virus (NiV) is a threat to global health security. On May 17, 2018, a 28 year old male presented to a private facility in Kozhikode district, Kerala, with encephalitis. The etiology was identified as NiV. We describe the characteristics of this outbreak and evaluate human to human transmission dynamics of NiV. Methods: We confirmed NiV via real-time PCR of specimens from throat swabs, blood, urine and CSF. The virus was sequenced using NGS and subjected to phylogenetic analysis. We conducted an epidemiological investigation to describe the outbreak and to elucidate transmission dynamics. Findings: During May 2 - 29, 2018, 23 cases were identified including the index case; 18 were lab- confirmed. This NiV was 97% similar to Bangladesh lineage. Median age of cases was 45; 15 (65%) were male. The median incubation period was 9.5 days (6 - 14 days); of the 23 cases, 20 (87%) had respiratory symptoms. The case fatality rate (CFR) was 91%; two cases survived. The index case acquired disease in the community, remaining transmissions were nosocomial. Risk factors for infection included close proximity (touching, feeding or nursing a NiV infected person), enabling exposure to droplet infection. Public health response included isolation of cases, contact tracing, and enforcing hospital infection control practices. Interpretation: This is the first recorded NiV outbreak in South India. Early laboratory confirmation and immediate public health response contained the outbreak. Rapid detection and response to outbreaks, and the culture of laboratory confirmation and infection control practices should be strengthened. Funding Statement: Department of Health Research, Indian council of Medical Research. (We thank adhoc research grants from Department of Health Research, Indian council of Medical Research for the VRDL (File No. 5/8/7/15/2010– ECD– I) and Nipah outbreak investigation (File No. VIR/1/2015/ECD- I) to Manipal Centre for Virus Research (MCVR), Manipal Academy of Higher Education.) Declaration of Interests: No potential conflict of interest relevant to this article was reported. Ethics Approval Statement: The outbreak investigation was reviewed and approved by Institutional Ethics Committee of Manipal Academy of Higher Education (File No. MAHE EC/009/2018).

Molecular characterization of neuraminidase genes of influenza A(H3N2) viruses circulating in Southwest India from 2009 to 2013.

Molecular characterization of neuraminidase (NA) gene of 25 influenza A(H3N2) virus isolates (2009-2013) archived at the Manipal Centre for Virus Research was carried out. The annual rate of amino acid substitutions in the N2 gene of influenza A(H3N2) virus isolates was 0.2-0.6%. Out of the 25 NA sequences analyzed, catalytic site mutations were observed in three isolates. Two of the mutations (D151G and E276G) were detected in functional catalytic residues, and an E227V mutation was detected in the framework residues. To the best of our knowledge, NA inhibitor resistance associated with the mutations E276G and E227V has not been reported. However, the mutation D151G, which is commonly associated with culturing of influenza A(H3N2) virus in Madin-Darby canine kidney (MDCK) cells, has been reported to result in a reduction in virus susceptibility to NA inhibitor drugs. Our study also detected mutations in antigenic residues. Some of the mutations (except D197G, K249E, A250T, S334C, and H347R/N) remained conserved in isolates of succeeding seasons. Antigenic residue mutations (D197G and S334C) have not been reported globally to date. The effect of these catalytic and antigenic mutant residues on drug and antibody binding was analyzed using three-dimensional structural analysis and biochemical assays. Antigenic variability of influenza A(H3N2) viruses is a major concern, and vaccine failures are mainly due to genetic variations in the HA gene. Our study documents that genetic changes in N2 occur at a slower rate, and this information is useful for the consideration and standardization of NA in influenza vaccines.

Circulating Genotypes of Dengue-1 Virus in South West India, 2014-2015.

Dengue is the most prevalent arboviral disease affecting humans, with tropical and subtropical regions having the highest risk of dengue virus infections. The clinical symptoms often range from mild fever to fever with hemorrhagic manifestations and shock. Since 2010, India has been witnessing a marked increase in the number of dengue virus serotype 1 (DENV-1) infections, apart from those of serotypes 2 (DENV-2) and 3 (DENV-3). The present study was undertaken to understand the circulating genotypes of DENV-1 in South West India, by sequencing the envelope gene of DENV-1 samples representative of the 2014-2015 time period. The extracted RNAs from dengue NS1 antigen ELISA-positive samples, archived at the APEX Referral Laboratory for Arboviruses (National Vector Borne Disease Control Programme, Manipal Centre for Virus Research), were used for the study. The currently circulating genotypes of DENV-1 identified from the study area belong to genotypes V and I. Multicentric laboratory-based studies analyzing the envelope gene of all dengue serotypes are required from endemic countries like India.

Oncolytic herpes viruses, chemotherapeutics, and other cancer drugs.

Oncolytic viruses are emerging as a potential new way of treating cancers. They are selectively replication-competent viruses that propagate only in actively dividing tumor cells but not in normal cells and, as a result, destroy the tumor cells by consequence of lytic infection. At least six different oncolytic herpes simplex viruses (oHSVs) have undergone clinical trials worldwide to date, and they have demonstrated an excellent safety profile and intimations of efficacy. The first pivotal Phase III trial with an oHSV, talimogene laherparepvec (T-Vec [OncoVexGM-CSF]), is almost complete, with extremely positive early results reported. Intuitively, therapeutically beneficial interactions between oHSV and chemotherapeutic and targeted therapeutic drugs would be limited as the virus requires actively dividing cells for maximum replication efficiency and most anticancer agents are cytotoxic or cytostatic. However, combinations of such agents display a range of responses, with antagonistic, additive, or, perhaps most surprisingly, synergistic enhancement of antitumor activity. When synergistic interactions in cancer cell killing are observed, chemotherapy dose reductions that achieve the same overall efficacy may be possible, resulting in a valuable reduction of adverse side effects. Therefore, the combination of an oHSV with “standard-of-care” drugs makes a logical and reasonable approach to improved therapy, and the addition of a targeted oncolytic therapy with “standard-of-care” drugs merits further investigation, both preclinically and in the clinic. Numerous publications report such studies of oncolytic HSV in combination with other drugs, and we review their findings here. Viral interactions with cellular hosts are complex and frequently involve intracellular signaling networks, thus creating diverse opportunities for synergistic or additive combinations with many anticancer drugs. We discuss potential mechanisms that may lead to synergistic interactions.

Prevalence of Hepatitis C Virus and its genotypes among a cohort of drug users in Kenya

Prevalence of hepatitis C virus and that of its main genotypes varies between the worlds geographic regions. The risk factors for infection with HCV include blood transfusion, tattoing and injecting drug use. To examine the prevalence of HCV and determine its main genotypes among a cohort of drug users in Kenya. A laboratory based study. Hepatitis research laboratory in the Centre for Virus Research at the Kenya Medical Research Institute, Nairobi. Three hundred and fourteen male and 19 female intravenous and non-intravenous drug users aged between 15-55 years. Seventy four (22.2%) out of 333 samples tested positive for anti-HCV. Sixty nine out of the 74 serum samples were assayed for HCV RNA and 38 (55.5%) were positive. The RNA positive samples were further subjected to sequencing and 19 (73%) of the samples were classified as genotype 1a, while seven (27%) samples were classified as genotype 4. Genotypes 2, 3, 5 and 6 were not identified in this study. These results demonstrate a high HCV infection prevalence among this cohort of drug users (22.2%) as compared to that of the general population, which is estimated to be 0.2-0.9%. The study also confirms the presence of at least two major genotypes among Kenyan drug users (genotypes 1 and 4).

New indirect immunofluorescence assay as a confirmatory test for human immunodeficiency virus type 1

Screening of blood and blood products for human immunodeficiency virus (HIV) is routinely performed using the enzyme-linked immunosorbent assay (ELISA), and the results confirmed by Western blot (WB). However, western blot is expensive and mostly performed in developed countries. A technique more superior or comparable to WB and adaptable to developing countries must be sought. In an effort to identify such a technique, this study determined the efficiency of indirect immunofluorescence assay (IFA) to detect antibodies to HIV-1. To determine the accuracy and sensitivity of an in-house immunofluorescence assay (IFA) to detect antibodies to HIV-1 in plasma. A comparative study to evaluate the performance of indirect immunofluorescence assay (IFA) and western blot (WB) techniques in the detection of antibodies to HIV-1. Kenya Medical Research Institute, Centre for Virus Research. The study was conducted between June and December 2001. The evaluation of IFA as a technique for detecting antibodies to HIV-1 utilized a total of 400 samples. For these samples, IFA was compared with ELISA and particle agglutination (PA) (manuscript under preparation). Of the 400 samples, there were discrepant results in the three assays in only 36 samples. IFA was compared with Western blot (WB) to confirm the true HIV-1 serostatus in these 36 plasma specimens. The IFA technique used acetone-fixed HIV-1 infected MOLT-4 cells in one spot on a Teflon coated slide and uninfected MOLT-4 cells alone in a second spot to asses non-specific fluorescence. Western blot was performed according to the instructions of the manufacturer. The sensitivity and specificity of IFA based on 36 plasma specimens tested was 71.4% and 100% respectively. All samples that were HIV seronegative by WB were also HIV seronegative by IFA. However, two (5.6%) samples were HIV seronegative by IFA but seropositive by WB. The data obtained show that IFA can be used as a primary confirmatory test in Kenya.

Low prevalence of hepatitis C virus infection in Lusaka, Zambia

Journal Article Low prevalence of hepatitis C virus infection in Lusaka, Zambia Get access H. Oshitani, H. Oshitani 1Virology Laboratory, University Teaching Hospital, Lusaka, Zambia Address for correspondence: H. Oshitani, MD, Virus Research Centre, Sendai National Hospital, 2-8-8 Miyagino Miyagino-ku, Sendai, 983, Japan. Search for other works by this author on: Oxford Academic PubMed Google Scholar F. Kasolo, F. Kasolo 1Virology Laboratory, University Teaching Hospital, Lusaka, Zambia Search for other works by this author on: Oxford Academic PubMed Google Scholar N.P. Luo, N.P. Luo 1Virology Laboratory, University Teaching Hospital, Lusaka, Zambia Search for other works by this author on: Oxford Academic PubMed Google Scholar M. Mpabalwani, M. Mpabalwani 1Virology Laboratory, University Teaching Hospital, Lusaka, Zambia Search for other works by this author on: Oxford Academic PubMed Google Scholar K. Mizuta, K. Mizuta 1Virology Laboratory, University Teaching Hospital, Lusaka, Zambia Search for other works by this author on: Oxford Academic PubMed Google Scholar N. Numata, N. Numata 2Department of Microbiology, Sendai Municipal Institute of Public Health, Sendai, Japan Search for other works by this author on: Oxford Academic PubMed Google Scholar H. Suzuki, H. Suzuki 3Virus Research Centre, Sendai National Hospital, Sendai, Japan Search for other works by this author on: Oxford Academic PubMed Google Scholar Y. Numazamki Y. Numazamki 3Virus Research Centre, Sendai National Hospital, Sendai, Japan Search for other works by this author on: Oxford Academic PubMed Google Scholar Transactions of The Royal Society of Tropical Medicine and Hygiene, Volume 89, Issue 4, July-August 1995, Page 380, https://doi.org/10.1016/0035-9203(95)90017-9 Published: 01 August 1995 Article history Received: 12 December 1994 Accepted: 17 January 1995 Published: 01 August 1995

Bordetella Pertussis and Upper Respiratory Tract Infection Viruses in Egyptian Children with Whooping Cough

ABSTRACT Isolates of Bordetella pertussis and upper respiratory tract infection (URTI) viruses from nasopharyngeal secretions were studied in 100 Egyptian children with whooping cough. Thirteen (13...

Ctenophyllus (Conothobius) orientalis sp. n. (Siphonaptera: Leptopsyllidae), a Parasite of Pikas in Northern India

Ctenophyllus (Conothobius) orientalis sp. n. is described from material collected from Ochotona roylei and Rattus rattoides in Himachal Pradesh and Uttar Pradesh, India. The species, as is characteristic of all members of the genus, parasitizes a species of Ochotona. It is characterized by the presence of spiniform setae on the frons and the shape of the modified abdominal segments in both sexes. During extensive collecting activities conducted by personnel of the Virus Research Centre, Poona, India, a number of new species of ectoparasitic arthropods have been amassed. Among them are some undescribed species of fleas which have contributed considerably to our knowledge of the range of certain genera. and the relationships of the various species. One of these, a new species of Ctenophyllus, is the subject of this study. Ctenophyllus (Conothobius) orientalis sp. n.

Haemaphysalis (Kaiseriana) anomala Warburton (Ixodoidea: Ixodidae) from India: Description of Immature Stages and Biological Observations

The immature stages of Haemaphysalis (Kaiseriana) anomala Warburton are described and characterized in relation to those of other species of the H. (K.) cornigera group. The previous single records of adults in widely scattered localities from Ceylon, Northeast India, and Southeast Asia are now expanded by relatively numerous data for adults, nymphs, and larvae from moist deciduous forests between 800 and 3,400 ft altitude in western Himalayan foothills and valleys of Uttar Pradesh and Himachal Pradesh. The temperature range in these forests is from 40 C in summer to 1 C, or rarely slightly less, in winter, and much of the total annual rain (125 to 300 cm) falls during the monsoons. Most adult ticks were taken during the monsoons; all immatures were collected in premonsoon and postmonsoon months. Nymphs and larvae were found feeding on a coucal, Centropus s. sinensis (Stephens) (Cuculidae), and rats (Rattus spp.). Adults were common on domestic cattle and buffalo. In the laboratory, the threehost type of life cycle was completed in about 4 months. Haemaphysalis (Kaiseriana) anomala Warburton was previously known only from 14 collections of adult specimens taken from humans, carnivores, and artiodactyls between 800 and 4,000 ft altitude in Northeast India, Nepal lowlands, and Southeast Asia, and it was suggested that these scattered data might be associated with certain peculiar biotopes (Hoogstraal, Kohls, and Trapido, 1967). A single collection from Ceylon has also been reported (Seneviratna, 1965). Owing to the little information regarding the widely distributed H. (K.) anomala, it is noteworthy to report the discovery of an ecological zone in the western Himalayan foothills of Uttar Pradesh and Himachal Pradesh where Received for publication 23 November 1971. * From Research Project MF12.524.009-3010, Bureau of Medicine and Surgery, Department of the Navy, Washington, D. C. The opinions and assertions contained herein are the private ones of the authors and are not to be construed as official or as reflecting the views of the Department of the Navy or of the naval service at large. The illustrations in this report were prepared under the auspices of Agreement 03-005-01 between the NIAID (NIH) and NAMRU-3. The specimen material was collected during an ICMR-sponsored survey of hematophagous arthropods in mountain regions of India, supervised by Dr. T. Ramachandra Rao, former Director or the Virus Research Centre. t Medical Zoology Department, U. S. Naval Medical Research Unit Number Three (NAMRU-3), c/o Spanish Embassy, Cairo, Egypt. t Virus Research Centre, Indian Council of Medical Research (ICMR), Poona, India. relatively numerous specimens could be collected. From this material and data we now describe the immature stages, characterize them in relation to other members of the H. (K.) cornigera group, and provide host, distribution, and ecological records, as well as details of the life cycle in the laboratory. Haemaphysalis (Kaiseriana) anomala Warburton Anomalous scissor-spurred haemaphysalid (Figs. 1-15) The adults were redescribed and illustrated, and the biological information available for this species was reviewed by Hoogstraal, Kohls, and Trapido (1967).

Dermacentor raskemensis (Ixodoidea: Ixodidae), Redescription and Notes on Ecology

Dermacentor raskemensis Serdyukova in Pomerantsev, 1948, was described on the basis of a single male from Raskem region of Turkmen SSR, without any host record. Adults of this species have now been found on domestic animals, mostly sheep and goats, rarely yak and zo (a hybrid of yak and local strain of cattle), in several localities between 2,440 and 4,000 meters altitude in Himachal Pradesh and Jammu and Kashmir states of India, and from West Pakistan and Afghanistan. The male is redescribed and the female described for the first time. Adults are compared with those of D. marginatus, D. niveus, D. pavlovskyi, and D. everestianus. The collection localities are mostly in dry mountainous regions with little or no vegetation, except some places which have forests. There is some cultivation along river valleys or mountain slopes in some areas. Dermacentor raskemensis Serdyukova in Pomerantsev, 1948, was described on the basis of a single male from Raskem region, southeastern Turkmen SSR, collected on 12 October 1899, without any host record (Pomerantsev, 1950). Because of inadequate descriptions and illustrations, and due to the presence of a number of closely related species in the region, the exact identity of D. raskemensis has remained confused (Pomerantsev, 1950; Arthur, 1960). During the last few years, many collections of D. raskemensis were made by the Virus Research Centre personnel in Jammu and Kashmir and in Himachal Pradesh, India. Additional material was available for study from West Pakistan and Afghanistan. The present descriptions are based on a study of all this material. Dermacentor raskemensis Serdyukova in Pomerantsev, 1948 Pomerantsev's Raskem dermacentor

Haemaphysalis (H.) obesa Larrousse (Ixodoidea: Ixodidae) from Northeast India and Southeast Asia: Description of Immature Stages and Biological Observations

The larva and nymph of Haemaphysalis (H.) obesa Larrousse, described here, are quite similar to those of the Javan H. (H.) hirsuta Hoogstraal, Trapido, and Kohls. The two species comprise a structurally specialized and biologically similar H. (H.) obesa group. H. (H.) obesa inhabits dense, humid lowland forests from West Bengal and Assam (India) in the west, through Thailand southwestward into Perlis (the northernmost state of Malaysia) and northeastward into the Republic of Vietnam. Known hosts of adults are medium and large-size forest-dwelling mammals, the monkey, bear, boar, deer, and domestic cattle, and probably the elephant. The single nymph recovered from a host was biting on the neck of a child. Adults, nymphs, and larvae were swept or picked from forest vegetation; they rest on the underside near the tips of leaves within 1.5 m from the ground. In laboratories in India and Thailand, the life cycle was completed in about 100-130 days. The obesa group presents several unique and numerous interesting areas for structural, functional, ecological, biological, and epidemiological research. Haemaphysalis (H.) obesa Larrousse has been known only from 12 adult specimens (Hoogstraal et al., 1966; Hoogstraal and Rack, 1967). From the present description of laboratory-reared immature stages, the close structural and phylogenetic relationship of this species and the Javan H. (H.) hirsuta Hoogstraal, Trapido, and Kohls is confirmed. Both species, which form the H. (H.) obesa group, inhabit dense, humid, lowland forests, where adults feed on large and medium-size mammals. Owing to unique adult setal arrangements circumspiracularly and on femur IV, and to unusual female external genital features (Hoogstraal et al., 1966), functional anatomy, behavior, and sexual processes of the H. (H.) obesa Received for publication 7 July 1970. * From Research Project MF12.524.009-3010, Bureau of Medicine and Surgery, Department of the Navy, Washington, D. C. The opinions and assertions contained herein are the private ones of the authors and are not to be construed as official or as reflecting the views of the Department of the Navy or of the naval service at large. The illustrations in this report were prepared under the auspices of Agreement 03-005-01 between the NIAID (NIH) and NAMRU3. t Medical Zoology Department, U. S. Naval Medical Research Unit Number Three, Cairo, Egypt, UAR. $ Medical Zoology Department, Niigata University School of Medicine, Niigata City, Japan. ? Virus Research Centre, Poona, India. The Virus Research Centre is maintained by the Indian Council of Medical Research. group should be exceptionally interesting to compare experimentally with other haemaphysalid groups. Larval and nymphal hosts in nature remain unknown. Ecological characteristics, known host preferences of adults, rapid life cycle, and ease of collecting and rearing these large ticks also serve to recommend this group as an outstanding candidate for rewarding biological and epidemiological studies in the field and laboratory. Materials and data for this report derive from a program funded by the Indian Council of Medical Research to Dr. T. Ramachandra Rao, Director (now retired) of the Virus Research Laboratory, Poona, for investigating the Himalayan area tick fauna of India (Dhanda and Bhat data); the German India Expedition, 1955-58 (Hoogstraal and Rack, 1967); a tick survey of Nakhon Ratchasima, Thailand, and laboratory studies in the Faculty of Tropical Medicine, University of Medical Sciences, Bangkok (Saito data); the Malayan Tick Survey, a collaborative study between the Institute for Medical Research (Kuala Lumpur), Zoology Department of the University of Maryland, and NAMRU-3 (Cairo) (Hoogstraal et al., 1969; Hoogstraal et al., in press); and continuing studies on the genus Haemaphysalis by the first author. Haemaphysalis (H.) obesa Larrousse Indo-Vietnam hirsute haemaphysalid (Figs. 1-15) Haemaphysalis obesa Larrousse, 1925, p. 302304, fig. 2, briefly described and illustrated $.

Haemaphysalis (Kaiseriana) ramachandrai sp. n. (Ixodoidea, Ixodidae), a Parasite of Man and Domestic and Wild Mammals in Northern India and Nepal

Haemaphysalis (Kaiseriana) ramachandrai sp. n. is described from field-collected and laboratory-reared adults and immature stages from forested lowlands in the Himalayan foothills of India and Nepal. Recorded hosts of adults are a human; sambar, barking, and chital deer; tiger and leopard; and domestic cattle, buffalo, and goats. Immature stages were taken in Nepal from the leopard, sambar, and chital. This species, a member of the H. (K.) bispinosa group of southern and eastern Asia and outlying islands, is characterized by small size, faint lateral grooves and scutal punctations, and reduction of certain coxal and trochanter spurs. Haemaphysalis (Kaiseriana) ramachandrai sp. n. is described from field-collected and laboratory-reared adults and immature stages from forested lowlands in the Himalayan foothills of India and Nepal. This distinctive, small member of the H. (K.) bispinosa group is dedicated to Dr. T. Ramachandra Rao, Director of the Virus Research Centre, Poona, in grateful appreciation of his many years of personal contribution to knowledge of vectors and vector-borne diseases of Asia and of his dedicated guidance to younger workers in these research areas. Haemaphysalis (Kaiseriana) ramachandrai sp. n. Ramachandra's Bispined Haemaphysalid

Ixodes himalayensis sp. n. (Acarina: Ixodidae) Parasitizing Small Mammals in Himachal Pradesh, India

Ixodes himalayensis sp. n. is described from female, nymph, and larva taken on rodents and a shrew from various localities in Himachal Pradesh, India. The species is related to I. redikorzevi Olenev, 1927, which is known to parasitize small mammals and birds in several localities in southwestern Russia, Greece, Bulgaria, Egypt, and Afghanistan. The new Ixodes species described below was collected by one of us (S.M.K.), during a survey of hematophagous arthropods in the Himalayan region of Himachal Pradesh, India. All the measurements in the descriptions are given in millimeters and were taken at the longest or the widest point. The illustrations were prepared from alcohol-preserved specimens with the aid of a camera lucida. Some of the larvae were mounted on slides in Hoyer's medium for studying the finer details and the chaetotaxy. Ixodes himalayensis sp. n. The Himalayan Small-mammal Ixodid (Figs. 1-16) Holotype Female, V.R.C. No. A-82035, Mus musculus, Palchan (altitude approximately 2,310 m), Kulu district, Himachal Pradesh, India, 3 October 1967, S. M. Kulkarni, deposited in the tick collection of the Virus Research Centre, Poona. Paratypes Eleven 9 9, 31 nymphs, 42 larvae, details as given in Table I. Paratypes deposited in collections of the Virus Research Centre, Poona; Indian Museum, Calcutta; British Museum (Natural History), London; and Rocky Mountain Laboratory, Hamilton, Montana. Female (Figs. 1-7) Body: Smallest specimen almost unengorged, oval, widest at level of coxae IV, 2.66 long, 1.51 wide. Largest specimen almost fully engorged, oblong, 5.11 long, 3.24 wide. Dorsal and ventral integumental areas thickly covered with moderately long white setae (Figs. 1, 2). Capitulum (Figs. 3, 4): 0.73 to 0.85 long, 0.46 to 0.56 wide. Basis capituli dorsally widest at juncture of palpi, converging gradually posteriorly to form well-developed sharp triangular cornua; posReceived for publication 13 January 1969. * The Virus Research Centre is maintained by the Indian Council of Medical Research. The Centre also receives a grant (3x4307) of the PL 480 Funds from NIH, USPHS, through the Indian Council of Medical Research. terior margin between cornua straight; porose areas large, clearly defined, subtriangular in outline. Basis capituli ventrally with well-developed posteriorly directed auriculae. Two pairs of minute posthypostomal setae. Palpi widest near apex of segment 2, greatest width about 0.16; segment 2 almost as long as 3, their combined length 0.54 to 0.61. Setae numerous, as illustrated (Figs. 3, 4). Hypostome (Fig. 5): Denticle-bearing portion 0.40 to 0.43 long; apex sharply pointed. Dental formula 3/3 in anterior half, thence 2/2 to base. Scutum: 1.12 to 1.40 long, 0.99 to 1.10 wide, widest near midlength. Scapulae sharp and pointed. Lateral carinae indistinct. Cervical grooves as shallow depressions, limited to anterior %, narrow and parallel anteriorly, wide and divergent posteriorly. Punctations large and distinct, moderately numerous, distributed all over scutum. Seta moderately long, white, distributed as illustrated (Fig. 1). Legs (Figs. 1, 2, 7): Moderately long. Coxa I with two spurs, outer small, triangular, inner long, tapering gradually to sharp points; II and III with well-developed triangular outer spur, and small distinct inner spur like projection; IV with only subtriangular outer spur. Trochanters lack spurs. Tarsus I 0.59 to 0.67 long, metatarsus I 0.31 to 0.41 long; tarsus IV 0.49 to 0.58 long, metatarsus IV 0.38 to 0.47 long. Pulvilli reach distal curvature of claws. Spiracular plate (Fig. 6): Outline circular, diameter about 0.22. Genital aperture: Situated at level between coxae III and IV. Genital groove rounded anterior to genital aperture, gradually divergent posteriorly, terminating short of posterior margin. Anal groove: Horseshoe-shaped, terminating short of posterior margin. Anus longitudinal slit, surrounded by two semilunar valves; each valve with three anal setae. Nymph (Figs. 8-14) Body: Shape as in female. Smallest specimen almost unengorged, 1.19 long, 0.64 wide; largest specimen almost fully engorged, 2.59 long, 1.65 wide. Dorsal and ventral integumental areas densely covered with long white setae. Capitulum (Figs. 10, 11): 0.25 to 0.28 long, 0.17 to 0.22 wide. Basis capituli dorsally and ventrally similar to female in outline, except that auriculae slightly smaller. Palpi widest near base