Typing of Plasmodium falciparum DNA from 2 years old Giemsa-stained dried blood spots using nested polymerase chain reaction assay.

A panel of 129 Giemsa-stained thick blood spots (TBS) confirmed for Plasmodium falciparum infection having different levels of parasite density were collected from a malaria endemic area. DNA was extracted and nested polymerase chain reaction (PCR) assay was performed to amplify P. falciparum DNA. Nested PCR assay successfully amplified P. falciparum DNA at a very low parasitaemia of ~10 parasites/µl of blood. Current PCR assay is very simple and can be used retrospectively to monitor the invasion and prevalence of different Plasmodium species in endemic areas.

Prioritizing areas for malaria control using geographical information system in Sonitpur district, Assam, India.

OBJECTIVE: To identify the malaria hot spots at health subcentre level in an endemic district using a geographical information system (GIS). The results will be useful for rapid retrieval of malaria information, and to prioritize malaria control efforts in identified hot spots. STUDY DESIGN: Extraction, analysis and synthesis of relevant data. METHODS: Malaria epidemiological data from 2006 to 2009 were analysed to determine the annual parasitic index, slide positivity rate, annual blood examination rate and Plasmodium falciparum percentage for each health subcentre in the district. Maps were produced using GIS, and integrated to identify the malaria hotspots. RESULTS: Out of 288 health subcentres, GIS identified 10 hot spots at extremely high risk of malaria and 14 hot spots at high risk of malaria. Malaria may flare up in these hot spots whenever favourable transmission conditions arise. Health authorities have been advised to establish control measures in these selected hot spots for timely prevention. CONCLUSION: There is a need for adequate monitoring and allocation of available resources for better interventions in the malaria hotspots. The GIS model used in this study can be used, even at village or cluster level, to pin point the malaria hot spots, and information can be updated and retrieved easily.

Studies on the ovitraps baited with hay and leaf infusions for the surveillance of dengue vector, Aedes albopictus in northeastern India.

Ovitraps baited with hay and leaf infusions were evaluated for enhancing the oviposition response of gravid females of the dengue vector Aedes albopictus. The egg density per trap (mean ± SEmean) was the highest with 30% infusions of Pennisetum grass hay (623.6 ± 41) and rice straw (580 ± 51.3), which corresponded to oviposition activity index (OAI) of 0.62. Infusions (5-50%) of mango and banana leaves with OAI ranging from -0.36 to 0.39 were not observed to enhance the oviposition response significantly over control. Rice straw and Pennisetum grass hay are available round the year in northeastern India and the use of these infusions can be a cost effective way to augment the ovitrap surveillance of dengue vectors.

Epidemiology and Risk Analysis of Malaria among Pregnant Women.

Malaria remains a complex problem during the pregnancy, which threatens > 35 millions pregnant women every year. Malaria pathogenesis in pregnancy results in accumulation of infected RBCs in the intervillous spaces causing severe alterations leading to the reduced materno-foetal exchanges. In this article we have revisited the current evidences of clinical implications and overall burden of malaria in pregnancy. Many adverse aftermaths including, low birth weight, intrauterine growth retardation, preterm delivery, stillbirth and anemia were found associated with malaria in pregnant women. Despite of worldwide comprehensive control programmes for malaria in pregnancy, the disease control has been a daunting task everywhere. Socio cultural, economical, lack of awareness and various logistic problems compound the disease in developing countries. Thorough evidence based information and estimates, education and awareness and strengthening of prevention programmes are needed urgently to achieve success in malaria control in pregnancy.

Insecticide resistance and detoxifying enzyme activity in the principal bancroftian filariasis vector, Culex quinquefasciatus, in northeastern India.

The insecticide resistance status of Culex quinquefasciatus Say (Diptera: Culicidae) to DDT and deltamethrin across army cantonments and neighbouring villages in northeastern India was investigated. In India, DDT is still the insecticide of choice for public health programmes. In military stations, pyrethroids, especially deltamethrins, are used for insecticide-treated nets (ITNs). Recent information on the levels of resistance to DDT and deltamethrin in mosquito populations of northeastern India is scare. Continued monitoring of insecticide resistance status, identification of the underlying mechanisms of resistance in local mosquito populations and the establishment of a baseline data bank of this information are of prime importance. Insecticide susceptibility assays were performed on wild-caught adult female Cx. quinquefasciatus mosquitoes to the discriminating doses recommended by the World Health Organisation (WHO) to DDT (4%) and deltamethrin (0.05%). Across all study sites, mortality as a result of DDT varied from 11.9 to 50.0%, as compared with 91.2% in the susceptible laboratory strain (S-Lab), indicating that Cx. quinquefasciatus is resistant to DDT. The species was found to be 100% susceptible to deltamethrin in all study sites except Benganajuli and Rikamari. Knock-down times (KDT) in response to deltamethrin varied significantly between study sites (P < 0.01) from 8.3 to 17.8 min for KDT(50) and 37.4 to 69.5 min for KDT(90). All populations exceeded the threshold level of alpha-esterase, beta-esterase and glutathion S-transferase (GST) established for the S-Lab susceptible strain, and all populations had 100% elevated esterase and GST activity, except Missamari and Solmara. Beta-esterase activity in Field Unit II (96.9%) was less than in any of the other populations. Benganajuli had the highest activity level for all the enzymes tested. There was a significant correlation between all enzyme activity levels and insecticide resistance phenotype by populations (P < 0.05). The results presented here provide the first report and baseline information of the insecticide resistance status of Cx. quinquefasciatus in northeastern India, and associated information about biochemical mechanisms that are essential for monitoring the development of insecticide resistance in the area.

Malaria situation in forest-fringed villages of Sonitpur district (Assam), India bordering Arunachal Pradesh during an outbreak.

BACKGROUND & OBJECTIVES: Epidemiological and entomological studies were undertaken in forest-fringed villages in Sonitpur district (Assam) bordering Arunachal Pradesh, India to assess the malaria situation. Blood smears (thick and thin) were collected from the fever cases through door-to-door survey. Both the blood smears stained with Giemsa were used for malaria parasite detection. Slide positivity rate (SPR) was recorded as 39.1% with predominance of Plasmodium falciparum (97.1%) infections. Children between 5 and 14 yr showed higher rates of infection. METHODS & RESULTS: Over all malaria prevalence was higher among the males (SPR 43.2%) than in females (SPR 34.5%). Adult mosquito collection was made using CDC miniature light-traps from dusk-to-dawn. Per-trap night density of mosquitoes in human dwellings and cattlesheds were 289 and 925, respectively. Anopheles minimus and An. dirus, the major vectors of malaria in Northeastern region of India formed 33.2% of the total vectors recorded. An. dirus could be recorded only from human dwellings. The results showed high malaria risk in the study villages. INTERPRETATION & CONCLUSION: High vector density with high parity rate, poor socio-economic conditions, lack of awareness, poor sanitation and congenial atmosphere for mosquito proliferation are aggravating the malaria situation in the study area.

Microencapsulation of Zanthoxylum limonella oil (ZLO) in glutaraldehyde crosslinked gelatin for mosquito repellent application.

Glutaraldehyde (GA) crosslinked gelatin (G) microcapsules containing Zanthoxylum limonella oil (ZLO) were prepared by coacervation technique. The effect of various parameters such as variation of oil-loading, gelatin concentration and degree of crosslinking on release rate of oil were studied. Scanning electron microscopy (SEM) was used to understand the surface characteristics of microcapsules. FTIR-results indicated the absence of any significant interaction between polymer and oil.

The drug sensitivities of Plasmodium falciparum in the Sonitpur district, Assam, India.

Falciparum malaria is an ongoing problem in the foothills of Northeast India. Evaluation of the drug sensitivities of P. falciparum was carried out in four endemic villages of the Sonitpur District of Assam, involving 218 cases who were tested in vivo over 35 days. Chloroquine resistance was detected at the RI level in 29 cases (13%) and RII level in 8 cases (4%). No RIII chloroquine resistant cases were detected in the study. RI resistance was observed in the age groups 6-10 years, 11-14 years, and 15 years and above in 16%, 17%, and 13%, respectively. RII level resistance was observed in 4% of all those groups combined. All the RI and RII resistant cases responded well to a single dosage of Metakelfin (sulfamethoxypyrazine I.P 1,500 mg and pyrimethamine I.P 75 mg).

Evaluation of botanicals as repellents against mosquitoes.

Repellent properties of three plant extracts--essential oil (steam distillate) of Zanthoxylum limonella (fruits), Citrus aurantifolia (leaf) and petroleum ether extract of Z. limonella (fruits) were evaluated as repellent against Aedes (S.) albopictus mosquitoes in mustard (Dhara) and coconut (Parachute) oil base under laboratory conditions. Three concentrations--10, 20 and 30% of the repellents were evaluated. Repellents in mustard oil afforded longer protection time against the bites of Aedes (S.) albopictus mosquitoes than those in coconut oil. At 30% concentration, 296-304 min protection time was achieved by the test repellents in mustard oil base while repellents in coconut oil exhibited 223.5-245 min protection time at the same concentration. Oil of Z. limonella gave the highest protection time against the bites of Aedes (S.) albopictus mosquitoes at all the concentrations than other herbal repellents tested both in mustard and coconut oil.

Field evaluation of herbal mosquito repellents.

Repellent properties of Zanthoxylum armatum DC. Syn. Z. alatum Roxb. (Timur), Curcuma aromatica (Jungli haldi) and Azadirachta indica (Neem) oils were evaluated against mosquitoes in mustard (Brassica sp.) and coconut (Cocos sp.) oil base and compared with synthetic repellent. Dimethyl phthalate (DMP) as standard. Timur and jungli haldi afforded better protection in the both the base at all the concentrations. Tepellents in mustard oil gave longer protection time than those in coconut oil. At 0.57 mg/cm2 concentration timur oil gave significantly higher protection both in mustard (445 min) as well as coconut oil (404 min) than the other repellents and DMP.

Evaluation of Methoprene (Altosid) and Diflubenzuron (Dimilin) for control of mosquito breeding in Tezpur (Assam).

Insect growth regulators (IGRs) namely, Isopropyl (E-E)-(RS)-11-methoxy-3,7,11-trimethyldodeca-2, 4-dinoate (Methoprene) and 1-(4-cyclophenyl)-3-(2,6-diflerobenzoyl) urea (Diflubenzuron) were evaluated against mosquito larvae in laboratory as well as in different breeding habitats in Tezpur, Assam. LC90 values of diflubenzuron against Culex quinquefasciatus and Aedes albopictus were 0.0022 and 0.0027 ppm respectively, while it was 0.0027 and 0.0022 ppm respectively in case of methoprene. However, LC50 values of both the IGRs were almost same in case of Ae. albopictus and Cx. quinquefasciatus (varies between 0.0009 and 0.0011 ppm). In case of methoprene, maximum mortality was observed in pupal stage though the exposure was given in all the cases to the III instar larvae. Field trials were conducted in cemented drains, small ponds and ditches. At 0.2 ppm (0.020 kg/ha) both diflubenzuron and methoprene were found to eliminate 92-96 per cent Culex and Anopheles larvae. Methoprene and diflubenzuron were found equally effective for control of mosquito breeding in different breeding habitats and provide better efficacy than conventional larvicides and biocides.

Laboratory and field evaluation of Bacillus thuringiensis and B. sphaericus against mosquito larvae.

Laboratory and field trials were carried out with two formulations of Bacillus thuringiensis and four strains of Bacillus sphaericus (B 42, B 64, B 87 and B 33) against mosquito larvae in different breeding habitats of Tezpur, Assam. LC90 of B. thuringiensis var israelensis (formulation Teknar) against Ae. albopictus, Cx. quinquefasciatus and Cx. gelidus were recorded as 0.443, 0.453 and 2.15 ppm respectively and LC90 of B. thuringiensis (Deltox: VCRC B-17) against Ae. albopictus, Cx. quinquefasciatus, Cx. gelidus and Cx. malayi were 8.414, 11.22, 5.24 and 6.761 ppm respectively. LC90 of B. sphaericus strains B 42, B 64, B 87 and B 33 against Cx. quinquefasciatus were 0.055, 0.115, 0.046 and 0.257 ppm respectively. At the dosage of 1 l/ha 87 per cent mortality was achieved after 24 hrs with Bti and it increased to 90-95 per cent at 1.5 l/ha. In polluted cemented drains 93-97 per cent kill of Cx. quinquefasciatus was observed at 2.5 l/ha. Out of four strains of B. sphaericus evaluated, strain B 87 was found to be the most effective as 87-96 per cent kill was achieved with only 0.1 kg/ha for Cx. quinquefasciatus, Cx. vishnui and A. vagus. For others 0.2 kg/ha dosage, eliminated 96-100 per cent Cx. vishnui gr. for B 42, 92-93 per cent for B 64 and 90-93 per cent for B 33 strain.

Mosquito survey in Tripura.

Vector survey carried out in Tripura revealed the presence of 17 species of anophelines with a density of 92.39 per trap night. Major malaria vector encountered was Anopheles philippinensis which formed 30% of the total collection and is exophagic in nature. Another efficient malaria vector An. minimus which formed 5% of the total collection prevails equally both indoors and outdoors. The high parity rate of An. philippinensis (59%) and An. minimus (63.9%) gives a strong indication about their vectorial status in Tripura.

Active malaria transmission in South Mizoram.

Anopheles dirus was incriminated as malaria vector with 1.12% sporozoite rate in Tlabong (Demagiri) subdivision, Mizoram. High parity rate of A. dirus (33.69%) and A. minimus (52.87%) further confirmed their vectorial status. Both the vectors were found highly susceptible to 4% DDT.

Studies on malaria and filariasis vectors in Kamorta and Great Nicobar Islands.

A total of 1401 adult mosquitoes mainly dominated by Anopheles sundaicus (40.18%) and Culex quinquefasciatus (57.31%) were collected from Kamorta and Great Nicobar Islands. It was revealed from human bait collection that both species prevailed almost all through the night (density 8.1/man hour) with a peak around midnight. Mixed breeding of A. sundaicus with A. vagus, Cx. quinquefasciatus, Cx. bitaeniorhynchus and Cx. tritaeniorhynchus was observed at many places. Adult A. sundaicus were found highly susceptible to DDT and larvae to Temephos (Abate). Cx. quinquefasciatus was resistant to DDT (31% mortality). High parity of A. sundaicus (40.42%) is an indication of the vectorial potential of this species to sustain malaria transmission in these islands.