Genetic polymorphism and amino acid sequence variation in Plasmodium falciparum GLURP R2 repeat region in Assam, India, at an interval of five years.

BACKGROUND: The R2 repeat region of GLURP has been reported as a good genetic marker for Plasmodium falciparum genotyping. Proper knowledge of the extent and nature of P. falciparum genetic diversity using highly immunogenic R2 repeat region in malaria-endemic areas is a crucial element to understand various aspects related to immunity acquisition and disease pathogenesis. METHODS: Population diversity of P. falciparum GLURP and amino acid sequence repeats in GLURP R2 region was studied in malaria-endemic Assam state, northeast India and compared at an interval of five years during 2005 (Group-A) and 2011 (Group-B). RESULTS: Of the 66 samples, a total of 55 samples showed positive PCR bands for GLURP R2 region and altogether ten types of alleles with size ranging from 501 bp to 1,050 bp (50 bp bin) were observed and coded as genotypes I-X. In Group-A (n = 29), 24 samples were found infected with single, four with double and one with triple P. falciparum genotype, while in Group-B (n = 26), single genotype was found in 23 samples, double in two samples and triple in one sample. Genotype IV showed significant increase (p = 0.002) during 2011 (Group-B). Genotypes I to V were more common in Group-B (62%), however genotypes VI to X were more frequently distributed in Group-A. The expected heterozygosity was found slightly higher in Group-A (HE = 0.87) than Group-B (HE = 0.85), whereas multiplicity of infection (MOI) in Group-A (MOI = 1.21 ± 0.49) and Group-B (MOI = 1.12 ± 0.43) did not display significant variation. The amino acid repeat sequence unit (AAU) DKNEKGQHEIVEVEEILPE (called 'a') was more frequent in the well-conserved part of R2 repeat region. CONCLUSION: The present study is the first extensive study in India which has generated substantial data for understanding the type and distribution of naturally evolved genetic polymorphism at amino acid sequence level in GLURP R2 repeat region in P. falciparum. There was decrease in the PCR amplicon size as well as the number of AAU [amino acid repeat unit] in Group-B displaying the bottleneck effect. The present study described a new type of AAU 'd' which varied from the other previous known AAUs.

Spatiotemporal distribution of dengue vectors & identification of high risk zones in district Sonitpur, Assam, India.

BACKGROUND & OBJECTIVES: Dengue is an arboviral disease of public health importance in many parts of India and recently many cases have been reported from northeastern India. Aedes mosquitoes, which are the vectors of dengue, are widely prevalent in the region. A study was initiated in Sonitpur district of Assam to understand the spatiotemporal distribution and seasonal prevalence of dengue vectors and to identify the high risk zones. METHODS: Ovitrap surveys were conducted in three randomly selected villages under each of the eight public health centres (PHC) in district Sonitpur of Assam, northeastern India during March 2011-February 2012. Three risk zones (high, medium and low) were identified on the basis of per trap density of Aedes mosquitoes. Meteorological data were collected to study the temporal distribution of dengue vectors. RESULTS: Aedes albopictus (99.3%) was the predominant dengue vector followed by Ae. aegypti (0.7%) recorded in the ovitraps. The highest vector density was observed during the post-monsoon (60.1±18 per trap) while the lowest during the winter (7.6±4.9 per trap) and the season-wise differences in the vector density were significant (P=0.005). Maximum temperature (correlation coefficient, r=0.45) and minimum temperature (r=0.408) showed the highest positive correlation with the vector density, whereas the number of rainy days showed high positive correlation (r=0.185) than the total rainfall (r=0.117). The high risk zone (Dekhiajuli, Behali, Bihaguri and Gohpur PHC) as indicated by the high larval densities of dengue vectors, 45.3±18, 42.1±22.3, 36.9±29.1, 35.3±22.6 per trap, respectively, was validated by dengue epidemiological data collected during 2012. INTERPRETATION & CONCLUSIONS: Yearlong monitoring of dengue vectors was done for the first time in this region. Monthly maximum temperature and the number of rainy days could be used for the prediction of larval density of Aedes mosquitoes. The identification high dengue risk zones would help in adopting targeted interventions for disease management in future.

Monitoring of malaria, Japanese encephalitis and filariasis vectors.

BACKGROUND: Vector monitoring in military stations would help in protecting the armed forces from vector borne diseases such as malaria, Japanese encephalitis and filariasis. METHODS: Adult mosquitoes were collected from four villages around a military station in India using light traps and the species composition was estimated. Insecticide susceptibility of disease vectors against DDT, deltamethrin and permethrin was established using WHO kits. RESULTS: The known malaria vectors constituted 4.9% of the total mosquito collections and Anopheles philippinensis/nivipes (2.05%) was the most abundant. Japanese encephalitis and dengue vectors constituted 25.3 and 0.05% whereas the known vectors of both Japanese encephalitis and filariasis formed 50.9%. The mean (±SEmean) of annual parasitic index, slide positivity and Plasmodium falciparum percentage among the civilian population during the study period were 1.46 ± 0.37, 1.65 ± 0.77 and 50.2 ± 10.7. The filariasis vector Culex quinquefasciatus was resistant to DDT with 65.4% mortality whereas the DDT resistance in the Japanese encephalitis vector Culex vishnui gr. with 91.9% mortality needs to be confirmed. All other species tested were susceptible to DDT, deltamethrin and permethrin. CONCLUSION: Targeted interventions are needed to reduce the disease burden and vector activity in the villages adjoining the military station. The use of insect repellents, bed nets and repellent impregnated uniforms by the troops should be ensured for protection from vector borne diseases.

DDT & deltamethrin resistance status of known Japanese encephalitis vectors in Assam, India.

BACKGROUND & OBJECTIVE: Japanese encephalitis (JE) outbreaks are common in Assam, northeastern State of India. Information on resistance in known JE vectors in the affected area is important for effective control measures. This study was undertaken to determine the species abundance of JE vectors endemic to Sibsagar district of Assam, and their susceptibility against DDT and deltamethrin. METHODS: Adult mosquitoes were collected using CDC light trap and aspirators from human dwellings from 13 endemic villages falling under three Primary Health Centres. Collected mosquitoes were identified and unfed female mosquitoes were used for DDT and deltamethrin sensitivity bioassay. The bioassay was performed following WHO protocol using standard susceptibility test kit. Knockdown time (KDT) was monitored at every 10 minutes intervals, whereas mortalities were recorded 24 h post-exposure. Vector density and resistance status were mapped using geographic information system (GIS) technique. RESULTS: A total of 7655 mosquitoes were sampled under three genera, i.e. Anopheles, Culex and Mansonia, and nine species, the JE vector Cx. vishnui group (31.78%) was the most predominant species, followed by Ma. uniformis (16.81%) and Ma. indiana (16.45%). All vector species were suspected to be resistant to DDT and sensitive to deltamethrin, except Ma. indiana, which was suspected to deltamethrin resistant. The KDT50 and KDT95 values of vector mosquitoes for DDT were significantly higher as compared to deltamethrin. The probit model used to estimate KDT50 and KDT95 values did not display normal distribution of percentage knockdown with time for all the vectors tested for DDT and deltamethrin, except for Ma. indiana for deltamethrin assay and Cx. gelidus for the DDT assay. INTERPRETATION & CONCLUSION: Differences in insecticide resistance status were observed between insecticides and vector species. The results of this study provided baseline data on insecticide resistance in known JE vectors of Sibsagar, Assam. The maps generated may allow better communication in control operations and comparison of changes in susceptibility status of these vectors over time.

Nested PCR detection of Plasmodium malariae from microscopy confirmed P. falciparum samples in endemic area of NE India.

The present study evaluates the performance of OptiMAL-IT test and nested PCR assay in detection of malaria parasites. A total of 76 randomly selected blood samples collected from two malaria endemic areas were tested for malaria parasites using microscopy and OptiMAL-IT test in the field. PCR assays were performed in the laboratory using DNA extracted from blood spots of the same samples collected on the FTA classic cards. Of the total of 61 field confirmed malaria positive samples, only 58 (95%) were detected positive using microscopy in the laboratory. Sensitivity, specificity, positive predictive value, negative predictive value and false discovery rate of OptiMal-IT in comparison to the microscopy were 93%, 83%, 95%, 79% and 5%, respectively. On the other hand, the sensitivity and specificity of PCR assay were 97% and 100%, respectively, whereas positive predictive value, negative predictive value and false discovery rate were 100%, 90% and 0%, respectively. The overall performance of OptiMal-IT and PCR assays for malaria diagnosis was 76% and 97%, respectively. PCR assay enabled the identification of infection with Plasmodium malariae Laveran, 1881 in four samples misidentified by microscopy and Plasmodium-specific antigen (PAN) identified by the OptiMAL-IT test. In addition to the standard methods, such PCR assay could be useful to obtain the real incidence of each malaria parasite species for epidemiological perspectives.

Physicochemical characteristics of habitats in relation to the density of container-breeding mosquitoes in Asom, India.

BACKGROUND & OBJECTIVES: Container-breeding mosquitoes, especially Aedes spp are vectors of diseases such as dengue and chikungunya. The abundance of these disease vectors in an area depends on the availability of container habitats and their physicochemical characteristics. The species composition of container-breeding mosquitoes in Asom, India was studied and the larval density was correlated with the habitat characteristics. METHODS: Natural and man-made water-holding containers in Sonitpur district of Asom were surveyed for the presence of mosquito larvae. The percent composition of container-breeding mosquitoes and container index were calculated. The physicochemical characteristics of breeding water such as pH, conductivity, salinity, total dissolved solids, turbidity and dissolved oxygen were measured. RESULTS: Aedes albopictus (93.7%) was the predominant species in the container-breeding habitats whereas Culex quinquefasciatus (2.77%), Armigeres subalbatus (2.26%), Ae. aegypti (0.76%), Toxorhynchites sp (0.4%) and Lutzia sp (0.11%) were recorded in relatively low numbers. The larval density (mean ± SEmean) of the container breeding mosquitoes ranged from 4.4 ± 1.8 to 15.4 ± 8.2, while the container index ranged from 1.58 to 5.68%. The mean (± SEmean) pH, conductivity, salinity, total dissolved solids, turbidity, and dissolved oxygen of water in the container habitats were 7.15 ± 0.11; 396.1 ± 58.5 µS/cm; 0.24 ± 0.04 ppt; 207.1 ± 30.4 mg/l; 32.3 ± 5.1 NTU; and 1.42 ± 0.12% respectively. The mosquito larval density in the container habitats was having significant negative correlation with the conductivity of breeding water (r = - 0.89; p = 0.003). Salinity, total dissolved solids and turbidity of water in the habitats were negatively correlated, whereas pH and dissolved oxygen were positively correlated with the larval density. INTERPRETATION & CONCLUSION: The studies indicated the predominance of Ae. albopictus in the container-breeding habitats and reiterated its importance as a potential vector of dengue and chikungunya in the region. The spread of Ae. aegypti, the principal vector of dengue, in the semi-urban areas probably through road transport is a matter of public health concern. The use of conductivity of breeding water as an index for the proliferation of container breeding mosquitoes in the region could be explored further.

Malaria incidence among paramilitary personnel in an endemic area of Tripura.

BACKGROUND & OBJECTIVES: Paramilitary operations along the Indo-Bangladesh border are adversely affected by malaria induced morbidity and mortality. Villages surrounding the paramilitary installations often serve as disease reservoirs. Malaria incidence in Tripura State Rifles (TSR) units in Dhalai District of Tripura was studied and the role of the village population in disease transmission was also assessed. METHODS: Mass blood surveys were carried out among TSR personnel and villagers during 2007 to 2009. Malaria diagnosis through blood smear examination and rapid detection kits was done, and percentage parasitaemia was determined. Activity of malaria vectors was monitored using CDC light traps. RESULTS: Slide positivity rates (SPR) in the neighbouring villages (51.4%) was significantly higher than that in TSR (27.7%) (P<0.0001). Malaria incidence in villages did not show seasonal variability while it was lowest during post-monsoon season in TSR (P<0.325; OR = 0.74). Per cent Pf parasitaemia was high in TSR (0.29) as compared to villagers (0.20) (P<0.0001). Anopheles minimus and An. dirus were the major malaria vectors observed. INTERPRETATION & CONCLUSIONS: Paramilitary and public health authorities should adopt targeted measures to reduce the malaria incidence in the villages surrounding the paramilitary installations as the village populations play a major role in disease transmission.

Malaria epidemiology along Indo-Bangladesh border in Tripura State, India.

Malaria epidemiological surveys were conducted in 16 villages along the Indo-Bangladesh border in Tripura, northeastern India. Insecticide resistance among malaria vectors and chloroquine resistance in the parasite were also studied along with monitoring of vector density using light traps. The epidemiological data indicated that malaria incidence was highest during June-July and lowest during November. Examination of blood smears collected through door to door surveys indicated slide positivity rate (SPR) of 25.2% and that Plasmodium falciparum was the predominant parasite (slide falciparum rate of 22.3%). The incidence rates of falciparum malaria varied significantly among the age groups (p<0.001) and 2-4 year olds were the most affected. Major malaria vectors recorded in light trap collections were An. dirus, An. minimus and An. philippinensis/nivipes. Chloroquine resistance studies indicated that treatment failure occurred in 35% of the cases and hence the use of artesunate combination therapy (ACT) was recommended for treatment of malaria in the area.

Molecular analysis of knock down resistance (kdr) mutation and distribution of kdr genotypes in a wild population of Culex quinquefasciatus from India.

OBJECTIVE: To investigate the presence of knock down resistance (kdr) mutation, its frequency distribution in the principal vector of bancroftian filariasis, Culex quinquefasciatus from northeastern India, and to relate kdr genotypes with susceptibility and/or resistance to DDT and deltamethrin in this vectors. METHODS: Adult female mosquitoes were collected by aspiration from human dwellings in two villages, Benganajuli and Rikamari, and two military establishments, Field Units I and II. Insecticide susceptibility tests were performed following WHO methods with 4% DDT and 0.05% deltamethrin. Molecular identification of kdr mutation and genotyping of kdr locus was performed by allele-specific PCR (AS-PCR) and direct sequencing in a subset of samples. RESULTS: Mosquitoes were resistant to DDT and showed 11.9-41.2% mortality, whereas the knock down bioassay for deltamethrin suggests complete susceptibility to this insecticide in all study sites except Benganajuli. The result of AS-PCR confirmed the presence of three genotypes: susceptible (SS), resistant (RR) and heterozygous (SR) in the population. Genotype frequencies at kdr locus for DDT-resistant individuals conformed with the Hardy-Weinberg proportion, whereas DDT and deltamethrin susceptible individuals differed significantly (P < 0.05). The efficacy of AS-PCR in detecting the correct genotype was not encouraging. CONCLUSIONS: This is the first report from India on kdr genotyping in C. quinquefasciatus, and it confirms the occurrence of kdr allele in this vector in northeastern India. This finding has serious implications for the filariasis control programmes in India.

Development of physiological resistance and its stage specificity in Culex quinquefasciatus after selection with deltamethrin in Assam, India.

The study investigated the development and stage specificity of physiological resistance to insecticides in a colony of Culex quinquefasciatus Say (Diptera: Culicidae) mosquitoes, which are vectors of bancroftian filariasis in India, after selection with deltamethrin. Resistance was selected by exposing the larvae to the concentration of deltamethrin that caused 50% mortality in the tested population (i.e., LC50). Under continuous selection pressure, the LC50 increased steadily in subsequent generations. The estimated LC50 for the F0 generation was 0.409 microg/L; the LC50 first displayed a substantial increase in the F5 generation (5.616 microg/L) and reached 121.902 microg/L in the F10 generation. The objective of this study was to establish a deltamethrin-resistant colony to develop a research programme that will study the evolution of physiological resistance patterns and stage-specific resistance responses in Cx. quinquefasciatus larvae and adults under laboratory conditions. An approximately 298-fold increase in resistance was recorded after 10 generations, as evidenced by the resistance ratio (RR50). The progress and effect of the selection pressure in the adult stage was monitored with the World Health Organisation (WHO) diagnostic test. The mortality, as observed using the WHO diagnostic test, declined significantly from the F5 generation (85%) onwards and the highest rate of survival (65%) was observed in the F10 generation.

Role of asymptomatic carriers and weather variables in persistent transmission of malaria in an endemic district of Assam, India.

BACKGROUND: Malaria transmission is perennial in the Assam-Arunachal Pradesh interstate border areas in the Sonitpur district of Assam, India. A yearlong study was carried out on the incidence of symptomatic and asymptomatic malaria and the role of asymptomatic malaria carriers in persistent transmission of the disease. The relationships between malaria incidence and weather parameters were also investigated. METHODS: Active and mass blood surveys were conducted on a monthly basis in Bengenajuli, Sapairaumari Pathar, and Nigam villages near the Assam-Arunachal Pradesh border. Epidemiological indices were estimated for malaria-positive cases. Multiple linear regression between monthly malaria incidence and monthly average temperature, and relative humidity along with monthly total rainfall was carried out. The known malaria vectors collected in CDC light traps were identified and recorded. RESULTS: Slide positivity rate (SPR) and Plasmodium falciparum percent (Pf%) for symptomatic malaria were 26.1 and 79.8, respectively. Prevalence of malaria vectors was observed throughout the year with varying density. Anopheles philippinensis/nivipes and A. annularis were predominant among the seven known vector species recorded currently. Asymptomatic parasitemia was detected throughout the year with SPR ranging from 4.8 to 5.3. Monthly rainfall with 1-month lag had the highest correlation (r=0.92) with SPR. The relationship between SPR and weather factors was established as SPR=-114.22+0.58 Tmin+1.38 RH+0.03 RF (R2=0.89; p=0.00). CONCLUSION: Low and relatively constant levels of asymptomatic parasitemia was present in the study area. High malaria vector density and presence of asymptomatic malaria parasite carriers were responsible for persistent malaria transmission in the region. This study concludes that passive detection and prompt treatment of asymptomatic carriers is essential for preventing persistent disease transmission. Rainfall along with some other weather variables may be used for predicting the malaria epidemics in the region. The predictive information could be useful to target resources more effectively.

Insecticide susceptibility and dengue vector status of wild Stegomyia albopicta in a strategically important area of Assam, India.

BACKGROUND: Dengue vector control programmes are facing operational challenges due to resistance against commonly used insecticides throughout the endemic countries. Recently, there has been appreciable increase in the dengue cases in India, however, no recent data are available on susceptible status of dengue vectors. We have studied the susceptibility level of St. albopicta to commonly used insecticides in India. Adult mosquitoes were tested for the presence of dengue virus. METHODS: St. albopicta larval bioassays were carried out to determine the lethal concentrations (LC10, LC50 and LC99) and the resistance ratios (RR10, RR50 and RR99) for temephos. Susceptibility to 4% DDT, 0.05% deltamethrin and 5% malathion was assessed following standard procedure. Knock-down times (KDT10, KDT50 and KDT99) were estimated and knock-down resistance ratios (KRR10, KRR50 and KRR99) were calculated. VectorTest™ dengue antigen assay was used to detect the dengue virus in the field collected mosquitoes. RESULTS: In larval bioassays, the RR ranged from 1.4 (for RR99) to 1.7 (for RR50), which suggested that the tested St. albopicta were susceptible to temephos. There was no deviation among the lethal concentration data from linearity (r2=0.61). Adult St. albopicta mosquitoes were resistant to DDT, while fully susceptible to deltamethrin and malathion. The knock-down values (KDT10, KDT50 and KDT99) obtained for DDT displayed straight line in log-dose-probit analysis and follow linear regression model. The KRR99 for DDT was 4.9, which indicated a 4.9 folds increase in knock-down resistance to DDT. However, for malathion and deltamethrin, the KRR99 values were 1.6 and 1.5 respectively suggesting that mosquitoes were knock-down sensitive. None of the mosquito pool was dengue virus positive. CONCLUSION: St. albopicta showed resistance to DDT and reduced sensitivity to deltamethrin and malathion. This data on insecticide resistance could help public health authorities in India to design more effective vector control measures. More dengue vector specimens need to be scanned to identify the potential dengue vector.

Pfcrt mutant haplotypes may not correspond with chloroquine resistance.

INTRODUCTION: Chloroquine resistance in Plasmodium falciparum is associated with mutations in pfcrt and pfmdr1 genes. The frequency distribution of pfcrt K76T and pfmdr1 N86Y mutations and their association with chloroquine susceptibility was studied in an endemic area along the Indo-Bangladesh border. METHODOLOGY: A single-arm prospective study of clinical and parasitological responses in P. falciparum malaria patients to chloroquine was conducted in vivo. PCR-RFLP assay was used to detect pfcrt K76T and pfmdr1 N86Y mutations in P. falciparum. The PCR products of pfcrt gene were sequenced,  translated and aligned for haplotyping. RESULTS: Out of 63 cases, 44 (69.8%) responded adequately to chloroquine treatment. Pfcrt K76T mutation was recorded in 100% of the treatment failure cases, whereas pfmdr1 N86Y mutation was found in 52.6% of the cases only. Early treatment failure (84.2%) occurred more frequently than late treatment failure (15.8%). Kaplan-Meier survival analysis showed that the probability estimate for treatment success after 7 and 15 days was 0.84 (95% CI = 0.72-0.92) and 0.70 (95% CI = 0.57-0.80), respectively. Sequence analysis of 72 to 76 pfcrt gene codons revealed the presence of two mutant (CVMNT, CVIET) and two wild (CVMNK, CVIEK) haplotypes. The mutant CVIET haplotype was predominantly distributed (42.1%). CONCLUSIONS: The presence of mutations in pfcrt K76T and pfmdr1 N86Y genes is not sufficient to explain the therapeutic efficacy of chloroquine to P. falciparum. Study suggests that pfcrt K76T mutant haplotypes are widely distributed and are spreading diligently, which needs to be taken into account in devising an antimalarial policy.

High prevalence of pfcrt K76T and mdr1 N86Y mutations in Sonitpur district of Assam, India.

Present study reports the prevalence and distribution of pfcrt K76T and mdr1 N86Y mutations in malaria endemic areas of Sonitpur district of Assam. Out of 163 individuals tested for malaria, 67 (SPR = 41.1 %) were detected positive for malaria infection using rapid detection kit in the field and PCR assay in the laboratory. Nested PCR-RFLP assay was used to amplify pfcrt K76T and mdr1 N86Y genes flanking the K76T and N86Y mutations. P. falciparum was most abundant (91.04 %) among the three Plasmodium species reported and its prevalence was significantly higher as compared to P. vivax and P. malariae (χ(2) = 150.76; p ≤ 0.0001; df = 2). Malaria was equally distributed among all the age groups and both the sexes. Hemoglobin contents in severe anaemic patients had a significant linear decreasing trend among patients with the increase in age (χ(2) = 4.33; p = 0.03), whereas non severe anaemic patients exhibited significant linear increasing trend among the patients with the increase in age (χ(2) = 18.38; p ≤ 0.0001). Pfcrt K76T mutation was recorded in 44 (72.13 %) isolates, whereas mdr1 N86Y mutation could be detected in 28 (41.79 %) isolates only. Only 32.7 % of the samples had both pfcrt K76T and mdr N86Y mutations. Number of pfcrt K76T mutant isolates was significantly higher than the wild type. However no significant difference was observed among the number of isolates with mdr1 N86Y mutant and wild isolates.

Antiplasmodial activity of certain medicinal plants against chloroquine resistant Plasmodium berghei infected white albino BALB/c mice.

In the present study of antimalarial efficacy, aqueous extracts of leaves and unripe fruits of Psidium guajava, leaves of Ocimum sanctum and leaves of Murraya koenigii are evaluated against Plasmodium berghei (chloroquine resistant NK65 strain) infected white albino BALB/c mice. A 7 days oral administration was adopted with different dosage viz., 350 mg, 750 mg and 1,000 mg/kg body weight as treatment schedule along with parasite (Group I) and drug control with Chloroquine, 50 mg/kg body weight (Group II). All the parts were extracted based on the decoction method, which is commonly seen among the villagers/tribes as their usual method of preparation of decoction for most of the ailments. The antimalarial activities were evaluated from the giemsa stained blood smears collected from different treated groups of mice used in this experiment. The antiplasmodial effect that is percent parasitaemia and percent suppression (values in parenthesis) showed by the treated groups of mice at 350 mg/kg b. wt. by the aqueous extracts of P. guajava leaves (Group III) was 19.8 ± 1.22 (73.7 %), P. guajava unripe fruits (Group IV) was 52.7 ± 2.19 (30.0 %), leaves of O. sanctum (Group V) was 64.0 ± 0.73 (15.1 %) and leaves of M. koenigii (Group VI) was 28.9 ± 0.81 (61.6 %) whereas at 750 mg/kg b. wt., it all showed 10.3 ± 0.7 (80.2 %), 26.3 ± 0.52 (65.1 %), 42.0 ± 0.47 (44.2 %) and 14.9 ± 0.46 (71.5 %) whereas at 1,000 mg/kg b. wt. dose, it all showed 9.2 ± 0.39 (85.8 %), 25.6 ± 0.40 (62.0 %), 41.8 ± 0.29 (35.5 %) and 14.0 ± 0.42 (76.9 %) respectively.

Chemotherapy and drug resistance status of malaria parasite in northeast India.

India reports the highest number of malaria cases in Southeast Asia, of which Plasmodium falciparum contribute more than half of the cases every year. North eastern states of India contribute only 3.96% of country's population but account for >10% of total reported malaria cases, 11% of Plasmodium falciparum cases and 20% of malaria related deaths annually. In India, chloroquine resistance was reported for the first time from northeast region and since then chloroquine treatment failure is being reported from many parts of the region. Increased chloroquine treatment failure has led to change of the drug policy to artemisinin combination therapy as first line of malaria treatment in the region. However, replacing chloroquine to artemisinin combination therapy has not shown significant difference in the overall malaria incidence in the region. The present review addresses the current malaria situation of northeastern region of India in the light of antimalarials drug resistance.

Polymerase chain reaction detection of human host preference and Plasmodium parasite infections in field collected potential malaria vectors.

This study was carried out to determine the human host preference and presence of Plasmodium parasite in field collected Anopheles mosquitoes among four villages around a military cantonment located in malaria endemic Sonitpur district of Assam, India. Encountered malaria vector mosquitoes were identified and tested for host preference and Plasmodium presence using PCR method. Human host preference was detected using simple PCR, whereas vectorial status for Plasmodium parasite was confirmed using first round PCR with genus specific primers and thereafter nested PCR with three Plasmodium species specific primers. Out of 1874 blood fed vector mosquitoes collected, 187 (10%) were processed for PCR, which revealed that 40·6% had fed on human blood; 9·2% of human blood fed mosquito were harbouring Plasmodium parasites, 71·4% of which were confirmed to Plasmodium falciparum. In addition to An. minimus, An. annularis and An. culicifacies were also found positive for malaria parasites. The present study exhibits the human feeding tendency of Anopheles vectors highlighting their malaria parasite transmission potential. The present study may serve as a model for understanding the human host preference of malaria vectors and detection of malaria parasite inside the anopheline vector mosquitoes in order to update their vectorial status for estimating the possible role of these mosquitoes in malaria transmission. The study has used PCR method and suggests that PCR-based method should be used in this entire malarious region to correctly report the vectorial position of different malaria vectors.

A cross sectional investigation of malaria epidemiology among seven tea estates in Assam, India.

In the present investigation, the epidemiology of malaria among seven tea estates of Nagaon and Udalguri districts of Assam, India has been described. A cross-sectional open study was carried out to understand the malaria epidemiology and associated risk factors among the tea tribes during March to September 2009. Out of 1,182 peripheral blood smears examined, 506 found positive for malaria (slide positivity rate, SPR = 42.8) with Plasmodium falciparum as predominant species. Dimakuchi tea estate was having highest SPR (P = 0.0275) and contributed more number of P. falciparum cases (P < 0.00001). Tea estates studied in both Udalguri and Nagaon districts were equally affected and the SPR recorded were 41.75 and 43.32% respectively. 154 malaria cases detected were having 'O' blood group but each blood group was found to have similar susceptibility of acquiring malaria infection (χ(2 ) = 3.603; P = 0.3076) and P. falciparum infection (χ(2 ) = 1.818; P = 0.6110). The SPR was highest among children more than 2 years of age group and variation in SPR among the age groups was statistically significant (χ(2 ) = 17.186; P = 0.0018). No gender biasing was observed in malaria distribution. Anemia was found associated with the infection among both the sexes. The findings suggest that tea estates are endemic for stable malaria transmission primarily due to P. falciparum and the prevalence rate decline with age, suggesting the development of protective immunity. Promising intervention measures could be able to reduce the malaria prevalence effectively in the study areas.