Molecular detection of Orientia tsutsugamushi in ectoparasites & their small mammal hosts captured from scrub typhus endemic areas in Madurai district, India.

BACKGROUND OBJECTIVES: Scrub typhus, caused by Orientia tsutsugamushi present in small mammals harbouring the ectoparasites. A study was undertaken to detect the pathogen present in small mammals and its ectoparasites in the scrub typhus-reported areas. METHODS: The small mammals (rodents/shrews) and its ectoparasites were screened for O. tsutsugamushi using nested PCR amplification of the groEL gene. Small mammals were collected by trapping and screened for ectoparasites (mites, ticks and fleas) by combing method. RESULTS: All the chigger mites collected were tested negative for O. tsutsugamushi . Interestingly, adult non-trombiculid mites ( Oribatida sp., Dermanyssus gallinae ), fleas ( Xenopsylla astia, X. cheopis, Ctenophalides felis and Ctenophalides sp.) and ticks ( Rhipicephalus sanguineus , R. haemaphysaloides ) screened were found to be positive for O. tsutsugamushi , which the authors believe is the first report on these species globally. Bandicota bengalensis with O. tsutsugamushi infection is reported for the first time in India. The O. tsutsugamushi groEL sequences from the positive samples were similar to the reference strains, Karp and Ikeda and phylogenetically clustered in clade IV with less evolutionary divergence. The blood samples of Rattus rattus , Suncus murinus and B. bengalensis collected from this area were tested positive for O. tsutsugamushi ; interestingly, the sequence similarity was much pronounced with their ectoparasites indicating the transmission of the pathogen to host or vice versa . INTERPRETATION CONCLUSIONS: The outcome of the present investigations widened our scope on the pathogens present in ectoparasites and rodents/shrews from this area. This will help to formulate the required vector control methods to combat zoonotic diseases.

A review of public health important fleas (Insecta, Siphonaptera) and flea-borne diseases in India.

Fleas (Insecta, Siphonaptera) are important vectors of plague and murine typhus in many parts of the world. Currently, about 2700 flea species were described in the world. The most common vector flea Xenopsylla cheopis is found throughout India, but X. astia, and X. brasiliensis are found less and limited in distribution associated with the domestic rats such as Rattus rattus, R. norvegicus, Mus musculus, and Bandicota bengalensis. Bubonic plague is a major flea-borne disease caused by the bacterial pathogen Yersinia pestis, transmitted from rats to humans via the rodent flea, X. cheopis. A major outbreak of plague and high mortality occurred in India. After 1966 with the 3 decadal intervals, plague cases occurred only during the year 1994 reported in 5 different states (Gujarat, Maharashtra, Karnataka, Uttar Pradesh, Madhya Pradesh and New Delhi and subsequently plague cases occurred during 2002 and 2004 after the one-decade interval in Himachal Pradesh (2002). Another outbreak of bubonic plague was reported in Dangud village, Barkhot tehsil, Uttarkashi district, Uttarakhand during October 2004. Ctenocephalides fleas are common in cats and dogs, which are the main vectors of bacteria rickettsiae, such as Rickettsia typhi, R. felis, R. conorii, and Bartonella henselae. Molecular and serological evidence also confirms the presence of R. typhi, R. conorii R. felis and B. henselae pathogens in cats and other fleas in India. Flea bites and flea-borne dermatitis are common in men and pet animals. Because of the re-emergence of the plague, updated information on fleas and flea-borne diseases are essential to control the flea vectors and flea-borne diseases in India. Hence, this comprehensive review updates the available information on fleas and fleas transmitted diseases in India.

Detection of Orientia tsutsugamushi in Novel Trombiculid Mite Species in Northern Tamil Nadu, India: Use of Targeting the Multicopy traD Gene.

Detection of Orientia tsutsugamushi DNA in a trombiculid mite chigger species suggests that it might be a potential vector of scrub typhus in an endemic area. Over a period of 20 mo, 85 rats were trapped, 57 had chiggers that were identified by standard morphometric techniques. The chigger pools were assessed by performing PCR assays targeting fragments of the single-copy genes 56 kDa type-specific antigen gene (TSA56) by nested PCR and the 47 kDa (htrA) quantitative real-time PCR (qPCR). The novel traD SYBR green assay that detects a multicopy gene was also performed. In total, 27 chigger pools were positive by traD qPCR, of which only 7 were positive by 47 kDa qPCR and in 3 of these, 56 kDa gene was amplified by nested PCR. Orientia tsutsugamushi-specific DNA was detected in Ascoschoengastia spp., Schoengastiella ligula, Leptotrombidium rajasthanense, Leptotrombidium deliense, and Leptotrombidium jayawickremei chigger pools. Therefore, they could be potential vectors of scrub typhus in Southern India. The three 56 kDa sequences belonged to TA716 genotype and Kato genogroup. Further studies are needed to confirm these chigger species as scrub typhus vectors in Northern Tamil Nadu.

Dengue infection among tribal population in the Nilgiris district, Tamil Nadu, India.

BACKGROUND & OBJECTIVES: Dengue emerged as an important public health problem in Tamil Nadu from 2000 onwards, reported in all the districts as an endemic disease of Tamil Nadu except Nilgiris district. So this study was carried out to understand the dengue epidemiology in Nilgiris district. METHODS: Block-wise study was made at the Nilgiris district. The clinicians at the Nilgiris Adivasi Welfare Association hospitals (NAWA) situated in Kotagiri, Kozhikarai and Primary Health Centers from Kunjpannai, Arayoor, and Soloor Mattam, examined and recorded symptoms and collected blood samples from the dengue-suspected patients. These samples were centrifuged at 4°C and stored. Serum samples (267 nos.) collected from dengue-suspected patients for two years period from 2014 to 2016 were screened for dengue infection. RESULTS: First year study conducted during 2014-15 showed 13 dengue positives (8.39%) mainly from Kotagiri block (9 nos. - 69.2%) and the second year study conducted during 2015-16 showed 12 dengue positives (10.71%) found mostly from Udagamandalam block (6 nos.- 50%). People belonging to 6 different tribes - Irular, Toda, Kota, Kurumba, Kattunaickan, and Paniya were found infected with dengue and more Irular positives were recorded in both the years (5 Irular-2014-15 & 11 Irular -2015-16). First year detected more female positives (92.3%) whereas the second year showed 5 males (41.7%) and 7 females (58.3%). INTERPRETATION & CONCLUSION: This study unearthed the hidden disease dengue to be prevalent among the tribal community and emphasized the need for the establishment of a permanent dengue surveillance system with improved disease diagnostics, to initiate effective vector control efforts to stop dengue transmission from this hilly region.

Changing paradigm in the epidemiology of Japanese encephalitis in a non-endemic region.

BACKGROUND & OBJECTIVES: Japanese encephalitis (JE) is a mosquito-borne zoonotic disease. The JE virus (JEV) does not cause any disease among its natural hosts and transmission continues through mosquitoes belonging to Culex vishnui subgroup. This study was aimed to investigate the prevalence of JEV in mosquitoes and humans in the Thanjavur district, a non-endemic region for JE, in Tamil Nadu, by using standard available assays. METHODS: : A sero-surveillance study was conducted in Thanjavur district among the normal rural school children in the 5-12 yr age group, during the JE season (October) and post-JE season (February) from 2011 to 2013 for the detection of JEV infection. Vector abundance studies were carried out from 2011 to 2014. JE seropositivity and its association between the seasons were analysed statistically. RESULTS: : The occurrence of JE infection among children aged 5-12 yr was very high in the study area. The infection rates for JE in two consecutive seasons for 2011-12 and 2012-13 were 32.2 and 65.2%, respectively. The Cx. tritaeniorhynchus sp. dominated the catch, and was majorly responsible for the transmission. There was a significant difference in the human infection rate compared to the years 1991-92 and 1992-93; and a marked decrease in the cattle to pigs ratio (123 : 1) compared to the studies in 1991-93. INTERPRETATION & CONCLUSION: : The study unearthed the prevailing situation of JE among children, who are at higher risk of developing the disease during the transmission season. The decrease in the cattle to pigs ratio might be the one of the reasons for increase in the JEV infection among the children population compared to 20 years before. This trend requires urgent attention as it could be prevented with effective surveillance systems and vaccines.

Emergence of dengue in the tribal pockets of Nilgiris district, Tamil Nadu.

As original tribal ways of living have morphed from a forest dweller existence, dengue is no longer an urban infection but is now also found in rural hilly areas. The spread of dengue is enhanced by the frequent movement of people to endemic areas where there is a vector mosquito presence. The impact of the virus is known to be great in the immunologically naive population. Our study reports on the threat of the dengue virus in these hilly areas.

Seasonal abundance & role of predominant Japanese encephalitis vectors Culex tritaeniorhynchus & Cx. gelidus Theobald in Cuddalore district, Tamil Nadu.

BACKGROUND & OBJECTIVES: Japanese encephalitis (JE) is the leading cause of viral encephalitis in Asia. The first major JE outbreak occurred in 1978 and since 1981 several outbreaks had been reported in the Cuddalore district (erstwhile South Arcot), Tamil Nadu, India. Entomological monitoring was carried out during January 2010 - March 2013, to determine the seasonal abundance and transmission dynamics of the vectors of JE virus, with emphasis on the role of Culex tritaeniorhynchus and Cx. gelidus. METHODS: Mosquito collections were carried out fortnightly during dusk hours in three villages viz. Soundara Solapuram, Pennadam, Erappavur of Cuddalore district. Mosquitoes were collected during dusk for a period of one hour in and around the cattle sheds using oral aspirator and torch light. The collected mosquitoes were later identified and pooled to detect JE virus (JEV) infection by enzyme linked immunosorbent assay (ELISA). RESULTS: A total of 46,343 mosquitoes comprising of 25 species and six genera were collected. Species composition included viz, Cx. tritaeniorhynchus (46.26%), Cx. gelidus (43.12%) and other species (10.62%). A total of 17,678 specimens (403 pools) of Cx. gelidus and 14,358 specimens (309 pools) of Cx. tritaeniorhynchus were tested, of which 12 pools of Cx. gelidus and 14 pools of Cx. tritaeniorhynchus were positive for JE virus antigen. The climatic factors were negatively correlated with minimum infection rate (MIR) for both the species, except mean temperature (P<0.05) for Cx. gelidus. INTERPRETATION & CONCLUSIONS: High abundance of Cx. tritaeniorhynchus and Cx. gelidus was observed compared to other mosquito species in the study area. Detection of JEV antigen in the two species confirmed the maintenance of virus. Appropriate vector control measures need to be taken to reduce the vector abundance.

Dengue vectors prevalence and the related risk factors involved in the transmission of dengue in Thiruvananthapuram district, Kerala, South India.

BACKGROUND & OBJECTIVES: A longitudinal, entomological and virological study was conducted from 2007 to 2010 in four dengue fever affected areas of Thiruvananthapuram district, Kerala to understand the risk factors involved in the dengue transmission. METHODS: Aedes surveys were carried out seasonally in the selected localities both indoors and peridomestic sites. Water holding containers were sampled for the presence of immature. Outdoor and indoor resting/landing mosquitoes were collected. Blood meal identification was performed by gel diffusion test and viral assay using the ELISA test. RESULTS: The species found were Aedes (Stegomyia) aegypti (Linn.), Ae. (Stegomyia) albopictus (Skuse) and Ae. (Stegomyia) vittatus (Bigot). Aedes aegypti and Ae. albopictus immature stages were also found during the study period. Aedes aegypti was the only prevalent species in the water-starved Vizhinjam, a rural coastal area with breteau index (BI) ranging from 40 to 271. Aedes albopictus was recorded in rest of the three surveyed localities­two urban and one rural ghat areas of Thiruvananthapuram district. INTERPRETATION & CONCLUSION: The vector control measures should be focused mainly on source reduction of water storage containers present in both outdoor (Ae. albopictus and Ae. vittatus) and indoor (Ae. aegypti). To achieve effective vector management, a public health response beyond routine larviciding or focal spraying is essential throughout the year.

Entomological investigations into an epidemic of Japanese encephalitis (JE) in northern districts of West Bengal, India (2011-2012).

BACKGROUND & OBJECTIVES: Japanese encephalitis (JE) is one of the most important arboviral diseases of human beings with outbreaks in many parts of Southeast Asia including India. We present the entomological findings of an outbreak occurred in northern part of West Bengal during 2011-2012 with special emphasis on the role of JE vectors in different seasons. METHODS: Adult mosquito collections were made with the help of mouth aspirators, aided by flash lights during day time resting inside human and animal habitations as indoor, and resting outside field grasses, bushes, underneath of culverts and bridges as outdoor, and in and around the pig enclosures and cattle sheds during dusk period in JE affected villages from Cooch Behar, Dakshin Dinajpur, Darjeeling and Jalpaiguri districts in North West Bengal. In all study villages, a long handled with enamel bowl dipper was used to obtain immature stages of mosquitoes from various breeding habitats. RESULTS: A total of 19 different types of mosquito breeding habitats were examined for vectors of JE. From these habitats, 23.7 per cent were positive for breeding during the study period. Overall, nine different species were recorded through emergence, but none was positive for JE virus when subjected for detection of virus. Adult mosquitoes of more than 50 per cent of the potential JE vector species obtained through dusk and the rest through indoor and outdoor collections in all seasons. Altogether, 27 different species were recorded. Most of these were JE vectors. INTERPRETATION & CONCLUSIONS: Our results showed that in addition to Cx. vishnui subgroup, detection of JE virus antigen in Cx. quinquefasciatus indicated the possible maintenance of JE virus in nature through poor vector mosquitoes throughout the year. Since, all potential vector species reported elsewhere in India were also found in this region and fluctuated in density in different seasons, a proper integrated vector control programme needs to be implemented to control JE transmission.

Description of a new species, Toxorhynchites (Toxorhynchites) tyagii (Diptera: Culicidae), from Nilgiri hills, Western Ghats, southern India.

Specimens of Toxorhynchites (Toxorhynchites) tyagii, sp. n., were collected from the fringe areas of Gudaloor town, Ooty in the Nilgiri hills at an altitude of 1000 m above sea level in Western Ghats ranges in southern India during October 2011 and from Darjeeling ranges in the northern hilly region of Raymatang TG in Jalpaiguri district, West Bengal, in eastern India during February 2012. The adults, pupa and fourth-instar larva of this species are described and illustrated to distinguish it from Toxorhynchites (Tox.) splendens and Toxorhynchites (Tox.) edwardsi, which are the closest allies of Tx. (Tox.) tyagii. Besides possessing remarkable distinguishing male genital characteristics, Tx. tyagii differs from Tx. splendens also by the presence of conspicuous yellow scale-patches over the wing root that extend to the scutellum, and differs from Tx. edwardsi by having midtarsomeres 3-5 all dark whereas in Tx. edwardsi tarsomeres 3 and 4 and a larger part of 5 are white.

Prevalence of Ascogregarina spp. in the container breeding Aedes albopictus from Chikungunya fever affected areas of Kerala State, India.

The prevalence of protozoan, Ascogregarina sp. had been determined in the container breeding mosquitoes, Aedes albopictus. Since, the cyst of Ascogregarina might play role in the maintenance of the Chik virus during silent period, the presence of Ascogregarina has gained importance in recent days. The prevalence was found to be 71.62.

A study on chikungunya outbreak during 2007 in Kerala, south India.

BACKGROUND & OBJECTIVE: The first chikungunya outbreak occurred in Kerala during 2006 affecting 14 districts, followed by another during May 2007 affecting almost whole of the State. Four of the worst affected districts viz, Pathanamthitta, Idukki, Kottayam and Thrissur were surveyed during 2007 to understand the magnitude of the problem of chikungunya fever, particularly clinical signs and symptoms. METHODS: A total of 1265 persons from 310 houses were surveyed door-to-door in 20 different localities representing four affected districts. The history and examination findings from 354 clinically diagnosed chikungunya cases were recorded. The symptoms recorded were fever, headache, myalgia, arthralgia, itch/rash, oedema, eye congestion, eye pain, oral ulcers, distaste, nausea, vomiting and haemorrhage. RESULTS: The major symptoms were fever (100%), headache (97.5%), arthralgia (99.4%) and myalgia (99.4%). A significant difference was observed in oedema, distaste, nausea and headache among different age groups and these symptoms were reported to be lower (12.2-89.8%) in younger age group than in older age group (90.4-100%). No genderwise difference was observed for any of the symptoms. In clinically diagnosed chikungunya cases higher age group (>35 years) found with higher rate of severity with symptoms of oedema, distaste, nausea and headache when compared with lower age group (1-35 yr). INTERPRETATION & CONCLUSION: Chikungunya invaded Kerala State for the first time in 2006 and continues to be a major vector borne disease in the State. The clinical symptoms in affected cases highlighted high fever, sever myalgia and prolonged arthralgia, with occasional history of skin itch/rash (petechiae).

Longitudinal studies of Japanese encephalitis virus infection in vector mosquitoes in Kurnool district, Andhra Pradesh, South India.

A 4-yr (2002-2006) entomological study was carried out in Kurnool district, Andhra Pradesh state, south India, to identify the mosquito vectors of Japanese encephalitis virus (family Flaviviridae, genus Flavivirus, JEV). In total, 37,139 female mosquitoes belonging five genera and 18 species resting on vegetation were collected in villages and periurban areas at dusk. Mosquito species composition and pattern of JEV infection in mosquitoes varied in periurban and rural areas. In periurban area, Culex gelidus Theobald was abundant, making up 49.7% of total catch followed by Culex tritaeniorhynchus Giles (44.5%). In rural area, Cx. tritaeniorhynchus was predominant, making up 78.9% of total catch followed by Culex quinquefasciatus Say (10.8%), Anopheles subpictus Grassi (7.1%), and Cx. gelidus (1.1%). In light trap collections, Cx. gelidus and Cx. tritaeniorhynchus predominated in periurban and rural areas, respectively. Of 50,145 mosquitoes screened JEV isolations were made only from Cx. gelidus and Cx. tritaeniorhynchus. Based on high abundance and frequent JEV isolation, Cx. tritaeniorhynchus was found to be the principal vector in both areas, whereas Cx. gelidus plays a secondary vector role in periurban areas only.

Entomo-epidemiological investigations on chikungunya outbreak in the Lakshadweep islands, Indian Ocean.

BACKGROUND & OBJECTIVE: An outbreak of chikungunya fever characterized by prolonged incapacitation in the Lakshadweep islands in Indian Ocean occurred in November 2006. We undertook the entomological and epidemiological investigations on the Chikungunya outbreak in the Lakshadweep islands, Indian Ocean. METHODS: Epidemic information in the affected places was obtained from the local hospital records. Entomological observations like larval survey and the adult resting/landing collections by the hand collection methods were individually conducted in the two affected islands Andrott and Kalpeni in November and December 2006. RESULTS: The main breeding sources of the mosquitoes were the rodent-devoured coconuts, coconut shells, areca nut soaking mud and plastic pots, discarded containers, grinding stones, metal containers and plastic containers. Aedes albopictus was pre-dominantly present in the Lakshadweep islands. INTERPRETATION & CONCLUSION: It is concluded that the routine campaigns need to be organized regularly within the community highlighting the potential breeding grounds of mosquitoes and the possible control methods. Source reduction strategies like destruction of coconut shells and rodent-devoured coconuts by burning or by burying them inside the ground. Ae. albopictus played major role as the vector mosquitoes responsible for the chikungunya transmission.