Sudhansu Sekhar Sahu

RDNA-ITS2 POLYMERASE CHAIN REACTION ASSAY FOR THE SIBLING SPECIES OF ANOPHELES FLUVIATILIS

Species-specific differences in the nucleotide sequences of the second internal transcribed spacer (ITS2) region of the ribosomal DNA (rDNA) were used to develop a diagnostic polymerase chain reaction (PCR) assay for two of the sibling species of the Anopheles fluviatilis complex, members of which are major vectors of malaria in central and northern parts of India. This assay consisted of a three primer reaction, which could amplify the DNA of both the species producing fragments of two distinct sizes, 350 bp for species X and 450 bp for species Y, respectively. The assay was found to be highly specific and sensitive.

Knowledge, attitude and practice on malaria: A study in a tribal belt of Orissa state, India with reference to use of long lasting treated mosquito nets

Local knowledge and practice related to malaria is important for the implementation of culturally appropriate, sustainable and effective interventions. In this context, to know peoples knowledge, attitude and practice on malaria and its prevention, a study was carried out in two districts viz., Malkangiri and Koraput of Orissa state in India, the former with ongoing insecticide treated mosquito nets (ITNs) programme and the latter without such programme (non-ITNs). Both qualitative and quantitative methods were used for data collection. The local concepts used for malaria describe only the biomedical symptoms of the disease although a few by meaning in local language reflect peoples misconceptions about the cause of malaria. About 63% of the respondents mentioned mosquito bite as the cause for this disease and 65% considered malaria as a serious problem. Qualitative data showed that people from remote villages seek treatment from traditional healers, Disharis. About 64% of the respondents stated that avoiding mosquito bites could prevent malaria. Majority (99%) of the people reported using personal protection measures to avoid mosquito bites. Although, majority of the people were aware of the cause and prevention of malaria (about 70% stated sleeping under mosquito net prevents malaria), a sizable proportion still had misconceptions and hence appropriate communication strategies should be developed and imparted alongside ITNs/LLINs distribution for a behaviour change to adopt such preventive measures. Since, the tribes are habituated to seek treatment from traditional healers; they could be involved in motivating people to use ITNs/LLINs to protect from mosquito bites and malaria.

Efficacy of aqueous suspension and granular formulations of Bacillus thuringiensis (Vectobac) against mosquito vectors.

The efficacy of aqueous suspension (AS) and granular (G) formulation of Bacillus thuringiensis var. israelensis (Vectobac) was tested against the immatures of mosquito vectors in the laboratory and under field conditions. Laboratory tests showed that the aqueous suspension was relatively more effective against Culex quinquefasciatus than Aedes aegypti and Anopheles stephensi, the respective LC(50) values being 0.046, 0.060 and 0.190 mg/l. In stream pools, with the application of Vectobac AS at 1.2 l/ha, more than 80% reduction in immature density of Anopheles larvae (Anopheles fluviatilis and Anopheles culicifacies) was observed for 2-8 days, and at 2.4 l/ha for 3.5-9.0 days. At the dosage of 7.0 kg/ha of the granular formulation, a reduction in immature density by more than 80% was observed for 2-9 days. In polluted habitats such as cesspits, U-drains and cement tanks, the effectiveness of Vectobac AS lasted for 1-4 days when applied at 1.2 and 2.4 l/ha, and Vectobac G was effective for 1-3 days at application rates of 7. 0 and 14.0 kg/ha against C. quinquefasciatus. There was no significant difference in the effectiveness between the two formulations and the two application rates.

Acceptability, willing to purchase and use long lasting insecticide treated mosquito nets in Orissa State, India

Long lasting insecticide treated nets (LLINs) that require no re-treatment have been advocated as an effective tool against malaria transmission. However, success of this community based intervention measure largely depends on its acceptability and proper usage by the target population, besides assuring access to bed nets. To determine the acceptability of LLIN, its usage and peoples willingness to buy the net, a study was conducted in two tribal districts viz., Malkangiri (with ongoing ITN programme) and Koraput (no ITN programme) of Orissa State, India. Both qualitative and quantitative data collection methods were used to collect information regarding the objective of the study. A total of 2457 LLINs (Olyset Nets) were distributed in the selected villages of these districts at free of cost. In the study villages of Malkangiri, 58% of the households had either ITNs (73%) or other types of mosquito nets aside from the LLINs and in the villages of Koraput, only 8% had other nets, as majority (96%) informed that buying nets from market was not affordable to them. Physical verification of the nets during the house visits revealed that 75.4% and 83% (in ITNs and non-ITNs villages, respectively) of the LLINs and 76% of the other nets (including ITNs) were used by the respondents, the night before the survey as nets were in a hanging position at the time of the visit. Majority of the respondents (76-98%) felt that reduction of mosquito bites as the main perceived benefit of using the LLINs. About 55% and 67% of the respondents from non-ITNs and ITNs areas, respectively, expressed their willingness to buy the LLINs. Among them, 76.8% and 94.7% offered to pay INR<100 for a net and also ready to buy it by cash payment. Social marketing of LLINs at a subsidized price or free supply to the deserving sections of people (socially/economically poor and/or under-privileged) and ensuring the availability of nets during harvesting season could encourage people to buy and use LLINs.

Reliability of light trap sampling for Anopheles fluviatilis, a vector of malaria.

The utility of light traps for sampling Anopheles fluviatilis, the major malaria vector in the hill tracts of Koraput district, Orissa, was assessed. A modified version of the CDC miniature light trap was used in the study. The data generated from light trap collections were analysed and compared with daytime resting, man-biting and night resting collections carried out in parallel. The number per trap recorded in cattle sheds was significantly higher than that recorded in human dwellings. A similar pattern was observed in hand catches of indoor resting population though the difference was not significant. The density recorded from light traps over time was found to have a significant positive linear association with corresponding indoor (r = 0.625; p = 0.0019) as well as outdoor (r = 0.603; p = 0.0029) diurnal resting densities. The proportion with different gonotrophic stages varied significantly between different types of collections. The age structure of A. fluviatilis did not differ significantly with the types of collection. As the light trap catches reflect seasonal fluctuations in the diurnal resting collections, they can justifiably be used in monitoring the relative density of A. fluviatilis in the place of cumbersome hand catch methods. Other uses of light traps are also discussed.

Comparative susceptibility of Anopheles subpictus from fresh and brackish water areas to Plasmodiumfalciparum infection

Anopheles subpictus from fresh and brackish water areas were fed simultaneously, on five volunteers, with Plasmodium falciparum gametocytes in their blood. The susceptibility to infection was compared. Infections were found in both waters, but to different degrees. The mean susceptibility rate among fed mosquitoes was 6.1% from the fresh water area and 17.4% from the brackish water area. The mean number of oocysts in the former was 3.4 +/- 3.2 and 4.2 +/- 4.32 in the latter. The mortality rate, up to day 9 post feeding, was 93.8% in fresh water form and 90.1% in brackish water form. Dissections of wild caught females showed that longevity of adults from both areas was shorter, 7-8 days. Despite a considerable proportion of mosquitoes being anthropophilic and susceptible to infection, their low longevity seems to be the limiting factor in their role in malaria transmission in these areas.

Glutathione S Transferase Activity in Indian Vectors of Malaria: A Defense Mechanism Against DDT

ABSTRACT Glutathione S transferases (GSTs) are multifunctional enzymes involved in detoxification of xenobiotic compounds in majority of the insect groups. Significance of insect GSTs is their elevated level of activity in association with insecticide resistance. This investigation was to explore the metabolic status of GSTs in two Indian DDT-resistant malaria vectors, Anopheles culicifacies and Anopheles annularis, and one DDT-susceptible vector, Anopheles fluviatilis. Malkangiri and Koraput districts of Orissa State, endemic for falciparum malaria and having a long insecticide spraying history, were the study areas. F1 progeny was raised from wild-caught females of the three vectors and used for biochemical assays to detect the GST-mediated DDT resistance mechanism. Results of the enzyme assay showed a significant 3-fold increase in GST activity in DDT-resistant An. annularis compared with its susceptible population. In DDT-resistant An. culicifacies, the median GST activity (71.8 µmol/min/mg) was almost the same as estimated in the DDT-resistant An. annularis (74.6 µmol/min/mg), suggesting that the GST activity estimated in An. culicifacies could be an elevated level for detoxification of DDT. Furthermore, the GST activity in DDT-resistant An. culicifacies and An. annularis was significantly higher than that in the DDT-susceptible An. fluviatilis, which had a GST activity of 20.0 µmol/min/mg. Also, the GST-mediated DDT detoxification was confirmed by comparing GST activity in wild-caught females with that in their F1 progeny.

Persistent foci of falciparum malaria among tribes over two decades in Koraput district of Odisha State, India

BackgroundKoraput, a predominantly tribe-inhabited and one of the highly endemic districts of Odisha State that contributes a substantial number of malaria cases to the India’s total. Control of malaria in such districts would contribute to change the national scenario on malaria situation. Hence, a study was carried out to measure the magnitude of malaria prevalence in the district to strengthen the malaria control activities.MethodsPrevalence of malaria was assessed through a sample blood survey (SBS) in seven randomly selected community health centres (CHCs). Individuals of all age groups in the villages selected (one in each subcentre) were screened for malaria infection. Both thick and thin smears were prepared from blood samples collected by finger prick, stained and examined for malaria parasites searching 100 fields in each smear. The results of a blood survey (n = 10,733) carried out, as a part of another study, during 1986–87 covering a population of 17,722 spread in 37 villages of Koraput district were compared with the current survey results. Software SPSS version 16.0 was used for data analysis.ResultDuring the current study, blood survey was done in 135 villages screening 12,045 individuals (16.1% of the total population) and among them, 1,983 (16.5%) were found positive for malaria parasites. Plasmodium falciparum was the major malaria parasite species accounted for 89.1% (1,767) of the total positives; Plasmodium vivax and Plasmodium malariae accounted for 9.3% (184) and 0.2% (5), respectively. Gametocytes were found in 7.7% (n = 152) of the positive cases. The majority of parasite carriers (78.9%) were afebrile. The 1986–87 blood survey showed that of 10,733 people screened, 833 (7.8%) were positive for malaria parasites, 714 (85.7%) with P. falciparum, 86 (10.3%) with P. vivax, 12 (1.4%) with P. malariae and 21 (2.5%) with mixed infections.ConclusionThe results of the current study indicated a rising trend in transmission of malaria in Koraput district compared to the situation during 1986–87 and indicated the necessity for a focused and reinforced approach for the control of the disease by improving people’s access to diagnosis and treatment and ensuring implementation of the intervention measures with adequate coverage and compliance.

Insecticide treated mosquito nets for malaria control in India-experience from a tribal area on operational feasibility and uptake

The study assessed the operational feasibility and acceptability of insecticide-treated mosquito nets (ITNs) in one Primary Health Centre (PHC) in a falciparum malaria endemic district in the state of Orissa, India, where 74% of the people are tribes and DDT indoor residual spraying had been withdrawn and ITNs introduced by the National Vector Borne Disease Control Programme. To a population of 63,920, 24,442 ITNs were distributed free of charge through 101 treatment centers during July-August 2002. Interview of 1,130, 1,012 and 126 respondents showed that the net use rates were 80%, 74% and 55% in the cold, rainy and summer seasons, respectively. Since using ITNs, 74.5-76.6% of the respondents observed reduction of mosquito bites and 7.2-32.1% reduction of malaria incidence; 37% expressed willingness to buy ITNs if the cost was lower and they were affordable. Up to ten months post-treatment, almost 100% mortality of vector mosquitoes was recorded on unwashed and washed nets (once or twice). Health workers re-treated the nets at the treatment centers eight months after distribution on a cost-recovery basis. The coverage reported by the PHC was only 4.2%, mainly because of unwillingness of the people to pay for re-treatment and to go to the treatment centers from their villages. When the re-treatment was continued at the villages involving personnel from several departments, the coverage improved to about 90%. Interview of 126 respondents showed that among those who got their nets re-treated, 81.4% paid cash for the re-treatment and the remainder were reluctant to pay. Majority of those who paid said that they did so due to the fear that if they did not do so they would lose benefits from other government welfare schemes. The 2nd re-treatment was therefore carried out free of charge nine months after the 1st re-treatment and thus achieved coverage of 70.4%. The study showed community acceptance to use ITNs as they perceived the benefit. Distribution and re-treatment of nets was thus possible through the PHC system, if done free of charge and when personnel from different departments, especially those at village level, were involved.

Bionomics of Anopheles minimus and An. fluviatilis (Diptera: Culicidae) in east-central India, endemic for falciparum malaria: human landing rates, host feeding, and parity.

ABSTRACT The host feeding of Anopheles minimus Theobald and An. fluviatilis James was studied in the villages of east-central India by conducting human landing collections between 1800 and 0600 hours at monthly intervals from May 2006 to July 2007. Four species of anopheline mosquitoes, An. minimus, An. fluviatilis, An. maculatus Theobald, and An. subpictus Grassi, were collected, the former two known vector species of malaria being the most abundant (97.1%). The An. minimus complex has been identified as sibling species A and An. fluviatilis complex as species S (90.9%) and T (9.1%). Both the species were prevalent throughout the year and obtained from landing collections indoors and outdoors. The average human landing density (HLD) of An. minimus and An. fluviatilis was 1.76 and 1.78 indoors and 1.71 and 1.56 per human per night outdoors, respectively. The HLD was relatively higher during the rainy season, although not significant in the case of An. fluviatilis. The human landing activity of An. minimus and An. fluviatilis occurred between 2000 and 0400 hours and peaked during 2300–0200 hours both indoors and outdoors.