Molecular identification of mosquito species in the Anopheles annularis group in southern Asia.

The Anopheles annularis group of subgenus Cellia Theobald (Diptera: Culicidae) includes five currently recognized species in southern Asia: An. annularis Van der Wulp, Anopheles nivipes (Theobald) and Anopheles philippinensis Ludlow, which are widespread in the region, Anopheles pallidus Theobald, which is known in Sri Lanka, India and Myanmar, and Anopheles schueffneri Stanton, which occurs in Java and Sumatra. Identification of the four mainland species based on morphology is problematic. In view of the fact that the three widespread species are variously involved in malaria transmission in different parts of the region, we developed a species-specific polymerase chain reaction assay based on rDNA internal transcribed spacer 2 (ITS2) sequences to facilitate entomological and epidemiological studies of the four species. The method proved to be reliable when tested over a wide geographical area.

Maintenance behaviour and long-lasting insecticide-treated nets (LLITNs) previously introduced into Bourapar district, Khammouane province, Lao PDR.

BACKGROUND: Long-lasting insecticide-treated nets (LLITNs) are expected to be an important advance in malaria control, but operational experience is still scarce. This study presents some operational findings concerning the introduction of Olyset LLITNs (Sumitomo Chemical Co., Ltd, Japan) in Laos. The study site, Bourapar district, a remote district at high risk of malaria, received Olyset nets during 1999-2000. After distribution of the nets the number of malaria cases in the district hospital decreased for a time, however it began to rise again a year after the intervention. To sustain the effect of the nets, net users were given instructions on maintenance and use. This study aimed to investigate the condition of Olyset nets and the maintenance behaviour of net users after 2-3 years of use, and to examine the associations between maintenance behaviour and the number of malaria episodes during the previous year. METHODS: Questionnaire interviews and inspections of nets were conducted at 240 households during February-March 2003. RESULTS: About 40% of the observed nets had holes/were torn, and the maintenance instructions had not been followed sufficiently. Households following the recommended washing frequency (38.2%) reported fewer malaria episodes during the past year, which demonstrates the importance of the recommended washing frequency in the effective use of the nets. CONCLUSIONS: Our study promotes the idea that, in addition to pursuing high coverage of LLITNs, more effort should be made to ensure that nets are kept in good condition in future LLITN programmes.

An open, randomized comparison of artesunate plus mefloquine vs. dihydroartemisinin-piperaquine for the treatment of uncomplicated Plasmodium falciparum malaria in the Lao People's Democratic Republic (Laos).

OBJECTIVE: To determine the efficacy and safety of oral dihydroartemisinin-piperaquine (DP, Artekin) in the treatment of uncomplicated Plasmodium falciparum malaria in southern Laos. METHODS: An open, randomized clinical trial of oral artesunate-mefloquine (AM) vs. DP in 220 patients with acute uncomplicated falciparum malaria in Savannakhet Province, Laos. RESULTS: The 42-day cure rates (95% CI), as determined by survival analysis and adjusted for reinfection, were excellent and similar for the two groups [99 (94-100)% and 100 (100-100)% for AM and DP, respectively]. The median (range) fever and parasite clearance times for the AM and DP groups were also similar [20 (4-63) h and 2 (1-4) days vs. 20 (7-57) and 2 (1-4) days, logrank P = 0.4 and 0.17, respectively]. There were more patients with at least one potential side effect following treatment in the AM group when compared with the DP group [64/110 (58%) vs. 48/110 (44%), respectively, P = 0.031]. CONCLUSION: Dihydroartemisinin-piperaquine did not have superior efficacy to AM for the treatment of uncomplicated falciparum malaria in Laos but was associated with fewer adverse effects.

Epidemiology of malaria in Attapeu Province, Lao PDR in relation to entomological parameters.

Surveys were conducted in malaria-endemic villages in the southern province of Attapeu, Lao PDR during various seasons over a 3-year period. All-night mosquito landing collections, blood surveys and a case-control study were conducted. Plasmodium falciparum was the predominant species, and slide positivity rates were higher during the transition/dry season compared with the wet season. Anopheles dirus A was found to be the primary vector, and sporozoite rates were highest during the transition/dry season. Anopheles dirus was found to be endophagic and endophilic. Not using insecticide-treated bed nets, houses close to breeding sites and sleeping away from home were risk factors associated with malaria.

Serological investigation of the prevalence of anti-dengue IgM and IgG antibodies in Attapeu Province, South Laos.

The prevalence of dengue antibodies was determined in the Attapeu region of South Laos with 225 blood samples collected from mostly febrile patients during the rainy season August - October 2001. An IgM capture ELISA was positive for one (0.4%) sample, while 177 (79%) samples were positive in an indirect IgG ELISA. Of the positive IgG samples, 20 (11.3%) were also positive on blood slides for Plasmodium falciparum. Dengue fever seems to be widespread in this area, but clinical dengue diagnosis remains difficult, especially in the first days of illness when physicians have to discriminate between dengue and other febrile illnesses.

Malaria transmission by Anopheles dirus in Attapeu Province, Lao PDR.

A study was carried out in four malaria-endemic villages in Attapeu Province, in the southern region of Lao PDR. All-night human landing collections were carried out in May, August, and October 2002, to determine malaria vectors. At the same time, mass blood surveys were also carried out in the same villages. Anopheles dirus was the predominant species in three of the study villages. Sporozoites were found only in An. drius from Phou Hom. However, in Beng Phoukham, An. dirus was positive for oocysts. The distribution of malaria cases was highest in Phou Hom and this correlated well with the vectorial capacity of An. dirus. The risk for infection from An. dirus was also high, at 0.99.

Endemic malaria in four villages in Attapeu Province, Lao PDR.

A study was conducted in four villages in Attapeu Province, Lao PDR in 2002 to determine malaria endemicity. The study villages were Mixay, Beng Phoukham, Phou Vong and Pier Geo. Mass blood surveys were conducted in May, August, and October. Finger prick blood was collected for thick and thin blood film as well as for dipstick. The slide positivity rate was highest in Phou Hom in October (41.7%). Plasmodium falciparum was the dominant species comprising more than 80% of the cases. As a whole, the distribution of malaria was similar among males and females. Children below 15 years accounted for a large percentage of the cases. The sensitivity of the optimal dipstick was 62.36 and the specificity was 61.7. Microscopy was taken as the gold standard. Anopheles dirus was found to be the main vector and the vectorial capacity correlated well with the cases.

The prevalence of Anopheles (Diptera: Culicidae) mosquitoes in Sekong Province, Lao PDR in relation to malaria transmission.

A longitudinal study was conducted on the prevalence of Anopheles in three malaria endemic villages in Sekong province, in the southern region of Lao PDR, from August 2000 to October 2001. All night, human landing collections took place in August and October 2000 and April and October 2001, and blood smears were taken for malaria parasites during the same period. Mosquitoes were tested for sporozoite antigen using enzyme-linked immunosorbent assay. In August 2000 (wet season) and April 2001 (dry season) the ovaries of the mosquitoes were examined for parity. A total of 16 species of Anopheles were caught in the study sites of which An. dirus A, An. maculatus sl and An. jeyporiensis were positive for sporozoites. The entomological inoculation rate (EIR) ranged from 0.06 to 0.25. There was a good correlation between EIR and vectorial capacity in the wet season, especially in Pai Mai where the prevalence of malaria was also high during the wet seasons (11.8 and 10.53). An. dirus A showed ambivalence in their choice of feeding as approximately 50% attacked man indoors and an equal proportion outdoors. An dirus A was the main vector in Pai Mai. The parous rate did not significantly differ between the wet and dry season, although it was higher in the dry season. In Takaio the parasite prevalence ranged from 8.7% (dry season) to 37.1% (wet season) and An. jeyporiensis was the vector, and the risk of infection was 0.85 in the dry season while 0.99 in the wet season. In Toumgno An. maculatus sl was the vector and infection was found only in August and October 2000. However, malaria prevalence ranged from 9.69 to 20.4% and was equally high in the dry season. Cattle were also present close to the houses in all the villages and this might be a contributory factor in the prevalence of malaria.

The efficacy of chloroquine for the treatment of acute, uncomplicated, Plasmodium falciparum malaria in Laos.

To assess the local efficacy of chloroquine for the treatment of acute, uncomplicated, Plasmodium falciparum malaria, children and adults from Sekong province (an area of Laos with a low intensity of transmission) were tested in a 28-day, in-vivo study. Complete data were collected from 88 of the 102 subjects enrolled between October 1999 and September 2000. After genotypic analysis to distinguish recrudescing infections from re-infections, 35 (39.7%, with a 95% confidence interval of 29.5%-50.7%) of these 88 patients were considered treatment failures. These results seriously question the use of chloroquine as the first-line treatment for P. falciparum malaria in the study area.