Role In Malaria Transmission Research Articles

The development and evaluation of a single step multiplex PCR for simultaneous detection of Anopheles annularis group mosquitoes, human host preference and Plasmodium falciparum sporozoite presence

The Anopheles annularis group mosquitoes, subgenus Cellia Theobald (Diptera: Culicidae), includes five recognized species: An. annularis Van der Wulp, An. nivipes Theobald, An. pallidus Theobald, An. philippinensis Ludlow and An. schueffneri Stanton. From these five, the three most common species found in Orissa were considered for this study because of their remarkable vectorial and behavioral variation and the important role they play in malaria transmission. To identify and understand their role in malaria transmission we developed a single multiplex PCR-based assay. This assay included the detection of human blood feeding habit and Plasmodium falciparum sporozoite presence. Of the 186 An. annularis mosquitoes collected, morphological character-based identification showed that 94 were An. annularis, 54 were An. philippinensis and 38 were An. pallidus. However, the multiplex PCR assay confirmed that 91 were An. annularis, 56 were An. philippinensis and 39 were An. pallidus individuals after adjustments were made for misidentified specimens in the morphological method. Anopheles annularis and An. philippinensis were found positive for human blood, and two samples of An. annularis species were positive for P. falciparum sporozoites. This one-step PCR-based method constitutes a very powerful tool in large surveys of anopheline populations.

Distribution, host preference and infection rates of malaria vectors in Mauritania

This study reports for the first time on the distribution, host preference and infection rates of malaria vectors in Mauritania. It was conducted during an outbreak of Rift valley fever. Three anopheline species were reported. An. arabiensis was the predominant species observed in all regions whereas An. pharoensis and An. funestus were observed along the south border in the Senegal River valley where extensive irrigation schemes are present. The distribution limits of anopheline species were observed from the Senegal River basin in the Trarza region up to the south limit of the Saharan desert in Tidjikja city. Overall, all An. funestus and An. pharoensis were fed respectively on human and ovine hosts whereas the mean anthropophilic rate of An. gambiae s.l. was 53%. A low Plasmodium falciparum infection rate was observed for species of the An. gambiae complex (0.17%) represented mainly by An. arabiensis. Because of the specific nature of this investigation, longitudinal studies are essential to better characterize the malaria vectors and their respective role in malaria transmission.

A pre-intervention study of malaria vector abundance in Rio Muni, Equatorial Guinea: Their role in malaria transmission and the incidence of insecticide resistance alleles

BackgroundFollowing the success of the malaria control intervention on the island of Bioko, malaria control by the use of indoor residual spraying (IRS) and long-lasting insecticide-treated nets (LLITN) was extended to Rio Muni, on the mainland part of Equatorial Guinea. This manuscript reports on the malaria vectors present and the incidence of insecticide resistant alleles prior to the onset of the programme.MethodsAnopheles mosquitoes were captured daily using window traps at 30 sentinel sites in Rio Muni, from December 2006 to July 2007. The mosquitoes were identified to species and their sporozoite rates, knockdown resistance (kdr) and acetylcholinesterase (AChE) sensitivity measured, to define the role of vector species in malaria transmission and their potential susceptibility to insecticides.ResultsA total of 6,162 Anopheles mosquitoes were collected of which 4,808 were morphologically identified as Anopheles gambiae s.l., 120 Anopheles funestus, 1,069 Anopheles moucheti, and 165 Anopheles nili s.l.. Both M and S molecular forms of Anopheles gambiae s.s. and Anopheles melas were identified. Anopheles ovengensis and Anopheles carnevalei were the only two members of the An. nili group to be identified. Using the species-specific sporozoite rates and the average number of mosquitoes per night, the number of infective mosquitoes per trap per 100 nights for each species complex was calculated as a measure of transmission risk. Both kdr-w and kdr-e alleles were present in the S-form of An. gambiae s.s. (59% and 19% respectively) and at much lower frequencies in the M-form (9.7% and 1.8% respectively). The kdr-w and kdr-e alleles co-occurred in 103 S-form and 1 M-form specimens. No insensitive AChE was detected.ConclusionAnopheles gambiae s.s, a member of the Anopheles gambiae complex was shown to be the major vector in Rio Muni with the other three groups playing a relatively minor role in transmission. The demonstration of a high frequency of kdr alleles in mosquito populations before the onset of a malaria control programme shows that continuous entomological surveillance including resistance monitoring will be of critical importance to ensure the chosen insecticide remains effective.

Species Composition and Distribution of Adult <I>Anopheles</I> (Diptera: Culicidae) in Panama

Anopheles (Diptera: Culicidae) species composition and distribution were studied using human landing catch data over a 35-yr period in Panama. Mosquitoes were collected from 77 sites during 228 field trips carried out by members of the National Malaria Eradication Service. Fourteen Anopheles species were identified. The highest average human biting rates were recorded from Anopheles (Nyssorhynchus) albimanus (Wiedemann) (9.8 bites/person/night) and Anopheles (Anopheles) punctimacula (Dyar and Knab) (6.2 bites/person/night). These two species were also the most common, present in 99.1 and 74.9%, respectively, of the sites. Anopheles (Nyssorhynchus) aquasalis (Curry) was encountered mostly in the indigenous Kuna Yala Comarca along the eastern Atlantic coast, where malaria case history and average human biting rate (9.3 bites/person/night) suggest a local role in malaria transmission. An. albimanus, An. punctimacula, and Anopheles (Anopheles) vestitipennis (Dyar and Knab) were more abundant during the rainy season (May-December), whereas An. aquasalis was more abundant in the dry season (January-April). Other vector species collected in this study were Anopheles (Kerteszia) neivai (Howard, Dyar, and Knab) and Anopheles (Anopheles) pseudopunctipennis s.l. (Theobald). High diversity of Anopheles species and six confirmed malaria vectors in endemic areas of Panama emphasize the need for more detailed studies to better understand malaria transmission dynamics.

Species Composition and Distribution of AdultAnopheles(Diptera: Culicidae) in Panama

Anopheles (Diptera: Culicidae) species composition and distribution were studied using human landing catch data over a 35-yr period in Panama. Mosquitoes were collected from 77 sites during 228 field trips carried out by members of the National Malaria Eradication Service. Fourteen Anopheles species were identified. The highest average human biting rates were recorded from Anopheles (Nyssorhynchus) albimanus (Wiedemann) (9.8 bites/person/night) and Anopheles (Anopheles) punctimacula (Dyar and Knab) (6.2 bites/person/night). These two species were also the most common, present in 99.1 and 74.9%, respectively, of the sites. Anopheles (Nyssorhynchus) aquasalis (Curry) was encountered mostly in the indigenous Kuna Yala Comarca along the eastern Atlantic coast, where malaria case history and average human biting rate (9.3 bites/person/night) suggest a local role in malaria transmission. An. albimanus, An. punctimacula, and Anopheles (Anopheles) vestitipennis (Dyar and Knab) were more abundant during the rainy season (May-December), whereas An. aquasalis was more abundant in the dry season (January-April). Other vector species collected in this study were Anopheles (Kerteszia) neivai (Howard, Dyar, and Knab) and Anopheles (Anopheles) pseudopunctipennis s.l. (Theobald). High diversity of Anopheles species and six confirmed malaria vectors in endemic areas of Panama emphasize the need for more detailed studies to better understand malaria transmission dynamics.

Identification of an eosinophil chemotactic factor from anopheline mosquitoes as a chitinase family protein

Anopheline mosquitoes play an essential role in malaria transmission. The mosquito salivates copiously when probing for the location of a blood vessel. We found that the saliva of anopheline mosquitoes has chemotactic activity for naive eosinophils or neutrophils. The major eosinophil chemotactic component in saliva was shown to be one of the chitinase family proteins. A similar chitinase family protein was found also in the midgut of the anopheline mosquito. Production of antibodies to the chitinase family protein was generally observed in the sera of residents of a malaria endemic area. Both Plasmodium falciparum-infected and uninfected individuals had antibodies to chitinases. These results suggest that the chitinase family protein in mosquito saliva contributes to eliciting an inflammatory response of eosinophils in the host skin followed by antibody production in the host.

Identification of four members of the Anopheles funestus (Diptera: Culicidae) group and their role in Plasmodium falciparum transmission in Bagamoyo coastal Tanzania

The role of Anopheles funestus group in malaria transmission was investigated in Bagamoyo coastal Tanzania, in the process of characterizing the area as a malaria vaccine testing site. Mosquitoes were sampled inside houses and multiplex PCR was used to identify 649 specimens. The following species were found: A. funestus s.s. (84.3%), A. leesoni (13.6%), A. rivulorum (1.5%) and A. parensis (0.6%). Multiplex PCR of 147 blood-fed specimens showed that over half (57.1%) of the identifiable blood meals were taken from human hosts, and human blood index in A. funestus and A. leesoni was 55% and 82% respectively. Plasmodium falciparum infection rate determined by nested PCR was 11% in A. funestus s.s. Although the abundance was low, 26 specimens of A. leesoni, two of A. rivolurum and one of A. parensis were found positive for P. falciparum. The presence of four A. funestus species in Tanzania emphasizes the relevance to define precisely their spatial and temporal distribution, specific behaviour, ecology and exact role in malaria transmission.

Biology and dynamics of potential malaria vectors in Southern France

BackgroundMalaria is a former endemic problem in the Camargue, South East France, an area from where very few recent data concerning Anopheles are available. A study was undertaken in 2005 to establish potential malaria vector biology and dynamics and evaluate the risk of malaria re-emergence.MethodsMosquitoes were collected in two study areas, from March to October 2005, one week every two weeks, using light traps+CO2, horse bait traps, human bait catch, and by collecting females in resting sites.ResultsAnopheles hyrcanus was the most abundant Anopheles species. Anopheles melanoon was less abundant, and Anopheles atroparvus and Anopheles algeriensis were rare. Anopheles hyrcanus and An. melanoon were present in summer, whereas An. atroparvus was present in autumn and winter. A large number of An. hyrcanus females was collected on humans, whereas almost exclusively animals attracted An. melanoon. Based on an enzyme-linked immunosorbent assay, almost 90% of An. melanoon blood meals analysed had been taken on horse or bovine. Anopheles hyrcanus and An. melanoon parity rates showed huge variations according to the date and the trapping method.ConclusionAnopheles hyrcanus seems to be the only Culicidae likely to play a role in malaria transmission in the Camargue, as it is abundant and anthropophilic.

Pattern of mitochondrial DNA variation between and within Anopheles stephensi (Diptera: Culicidae) biological forms suggests extensive gene flow

Anopheles stephensi Liston is the most prevalent anopheline species and plays an important role in malaria transmission in Indian subcontinent and Middle East including southern parts of Iran. It exists as three biological forms; “type”, “intermediate”, and variety mysorensis. The type form is reported to be an efficient vector of urban malaria, whereas mysorensis and intermediate are considered to be rural species and poor vectors. Moreover, differences in cuticular hydrocarbon and chromosomal characters have been described between urban and rural forms. However, the genetic structure of the biological forms remains unclear. This study was conducted to determine the genetic structure of the An. stephensi biological forms in south of Iran where all three forms are present. Live specimens were collected from the field and transferred to insectaries, reared, and identified based on egg morphological characters. Genetic structure of the biological forms was studied using PCR-RFLP of 1512 bp of mitochondrial DNA (mtDNA) cytochrome oxidase subunit I and II (COI–COII) and sequence of about 712 bp of COI and 562 bp of COII genes. Sequence analysis showed that except for a few substitutions in COII, all three forms and populations were nearly identical. The high homology of COI and COII sequence of An. stephensi forms indicates extensive gene flow between populations and forms in the region. This data will serve as first report on the sequence of mDNA COI–COII of biological forms of An. stephensi, which could be used as a diagnostic tool to identify vector/non-vector, gene flow, and geographical exchanges.

HOW RELIABLE IS THE HUMERAL PALE SPOT FOR IDENTIFICATION OF CRYPTIC SPECIES OF THE MINIMUS COMPLEX?

The Anopheles minimus Complex Theobald (Diptera: Culicidae) is composed of the 3 sibling species A, C, and E. The malaria vectors An. minimus A and C are distributed over the Southeast Asian region, whereas species E is restricted to the Ryukyu Japanese Islands. Because species A and C can be sympatric and present specific behaviors and have a role in malaria transmission, it is important to differentiate them. The literature mentioned the presence of a presector pale spot on the wing costa of An. minimus A, whereas species C may exhibit both presector and humeral pale spots. However, the reliability of their diagnostic power has not been established over large temporal and geographic surveys. From the analyses of 9 populations throughout Southeast Asia, including published data and field populations from 2 sites in Thailand, we showed that the wing patterns present spatial and temporal variations that make these two morphological characters unreliable for the precise identification of An. minimus A and C. Therefore, molecular identification remains the most efficient method to obtain an unambiguous differentiation of these 2 species. Correct species identification is essential and mandatory for any relevant study on the Minimus Complex and for the application of successful control strategies.

Molecular variation, systematics and distribution of the Anopheles fluviatilis complex in southern Asia

Species of the Anopheles fluviatilis James and Anopheles minimus complexes (Diptera: Culicidae) are difficult to distinguish morphologically. Members of the two complexes have been confused and, consequently, their distributions and roles in malaria transmission are uncertain. We identified numerous mosquitoes from China, Thailand, Vietnam, India, Nepal, Pakistan and Iran by single-strand conformation polymorphism (SSCP) and/or sequencing, and analysed the variation in the 28S D3 region of ribosomal DNA for members of the Minimus Subgroup and also the internal transcribed spacer region 2 (ITS2) for members of the An. fluviatilis complex. The D3 region is highly conserved between taxa and therefore could serve as a standard for molecular diagnosis of the subgroup members. D3 sequence, bionomics and malaria transmission data provide further evidence that An. fluviatilis S in India is conspecific with An. minimus C in South-east Asia. An. fluviatilis T has three ITS2 haplotypes (designated T1, T2 and Y) and its distribution in India, Nepal, Pakistan and Iran is confirmed. An. fluviatilis U is well defined on cytogenetic grounds in Uttar Pradesh, India, but is very close to An. fluviatilis T and the two species may hybridize in some regions. Variant ITS2 sequences suggest the possible existence of two additional taxa within the An. fluviatilis complex, one in Iran and another in India, provisionally designated An. fluviatilis forms V and X, respectively. The distributions of members of the An. fluviatilis and An. minimus complexes in south-central Asia are summarized.

RE-EMERGENCE OF ANOPHELES FUNESTUS AND ITS POSSIBLE EFFECT ON MALARIA TRANSMISSION ON MAYOTTE ISLAND, INDIAN OCEAN

Malaria constitutes a major public health problem on Mayotte Island, with the major vector being Anopheles gambiae s.s. The seeming disappearance of An. funestus, the 2nd vector on the island, was observed 15 years ago. Now, the presence of An. funestus during the biting collections performed in November and December 2004 confirms the re-emergence of this vector, which plays an important role in malaria transmission with An. gambiae.

Vecteurs de paludisme : du terrain à la génétique moléculaire Recherches en Afrique

Only about 60 Anopheline species transmit malaria among more than 3,000 mosquito species recorded in the world. In Africa, the major vectors are Anopheles gambiae,An. arabiensis, An. funestus, An. nili and An. moucheti. They all belong to species complexes or groups of closely related species that are very difficult to set apart on morphological grounds, but which may have highly variable behaviours and vectorial capacities. Understanding this complexity is of major importance in vector control programs or for implementing any public health intervention program such as drugs or vaccine trials. Among the seven species of the complex,Anopheles gambiaes.s. shows a huge chromosomal polymorphism related to adaptation to specific natural or anthropic environments, from equatorial forested Africa to dry sahelian areas. Recent studies conducted in West and Central Africa suggest an incipient speciation into 2 molecular forms provisionally called M and S. A similar evolutionary phenomenon is observed in An. funestus, in which sympatric populations carrying specific chromosomal paracentric inversions showed restricted gene flow. Distribution of species from An. nili group and An. moucheti complex is restricted to more humid regions of Africa. However in some areas these species play the major role in malaria transmission. Comprehensive knowledge of transmission cycles and of behavioural and underlying genetic heterogeneities that exist within and among natural vector populations will thus benefit the whole area of malaria control and epidemiology. Molecular and genetic studies, as well as in depth monitoring of vector biology, have been recently facilitated by advances in functional and comparative genomics, including recent publication of the nearly complete genome sequence of An. gambiae. Challenge for the next years is to answer to the very simple question: why is an insect a vector?

Identification of three members of the Anopheles funestus (Diptera: Culicidae) group and their role in malaria transmission in two ecological zones in Nigeria

The role of the Anopheles funestus group in malaria transmission was investigated in two ecological zones in Nigeria. Sampling was carried out at four sites each around Ibadan (forest) and Ilorin (savanna). Human landing catches were supplemented with indoor and outdoor resting collections. PCR was used to identify 1848 A. funestus group mosquitoes to species level (749 in the savanna, 1099 in the forest) and three species were identified. In the forest, A. funestus s.s. predominated (55.4%), followed by A. rivulorum (27.6%) and A. leesoni (17.0%). Anopheles funestus was found mostly indoors. Anopheles rivulorum and A. leesoni predominated in outdoor collections (P<0.001). Only Anopheles funestus s.s. was found in the savanna. ELISA analysis of 803 blood meal-positive specimens showed that over half of the blood meals were taken from humans in both ecotypes. The human blood index in A. funestus from the two study areas was similar. Anopheles funestus s.s. was the only species found positive for Plasmodium falciparum using ELISA, with overall infection rates of 2.3% and 1.0% in the forest and savanna respectively. The presence of three A. funestus species in Nigeria emphasizes the desirability of correct species identification within a malaria vector control programme.

POLYMERASE CHAIN REACTION DIAGNOSIS AND THE CHANGING PATTERN OF VECTOR ECOLOGY AND MALARIA TRANSMISSION DYNAMICS IN PAPUA NEW GUINEA

The ecology and behavior of most of the 11 known members of the Anopheles punctulatus group remain unresolved and only the morphologic species An. farauti, An. koliensis, and An. punctulatus are known as vectors of malaria in Papua New Guinea. Of 1,582 mosquitoes examined morphologically, 737 were identified as An. farauti s.l., 719 as An. koliensis, and 126 as An. punctulatus. All specimens identified morphologically as An. punctulatus were shown to be An. punctulatus by polymerase chain reaction-restriction fragment length polymorphism analysis, but the An. farauti and An. koliensis morphotypes consisted of three or more species including An. farauti s.s., An. farauti No. 2, and An. farauti No. 4. The biting cycles and role in malaria transmission of some of these species are described here for the first time. We also show evidence that An. koliensis could be a sub-complex of two or more species. The epidemiologic implications of our findings are discussed.

Distribution of members of Anopheles quadrimaculatus say s.l. (Diptera: Culicidae) and implications for their roles in malaria transmission in the United States.

The Anopheles quadrimaculatus s.l. (Say) complex consists of at least five species distinguished by distribution, genetic incompatibility, and allele frequencies. However, the distributions of the members have only been described by collection locations. Building on this information and environmental data, preliminary predictions of their distribution were produced using a genetic algorithm and point occurrence data. Based on resulting predicted border areas and undersampled regions, we obtained and analyzed additional geo-referenced specimens and compared their distribution with our preliminary predictions. We found good agreement between the preliminary predictions and the subsequent collections, regardless of the fact that additional specimens were deliberately sought from areas most likely to reveal inconsistencies. Final predicted distributions describe widespread distribution of A.quadrimaculatus throughout the eastern United States. A. maverlius and A. smaragdinus have similar predicted ranges limited to the southeastern United States. The predicted ranges of the sister taxa A. diluvialis and A. inundatus were similar to one another along the southeastern coast even though they seem to be allopatric. The historical role of A. quadrimaculatus s.l. in transmission of malaria was also examined. We conclude that A. quadrimaculatus s.s. was the only species of the complex capable of vectoring malaria in the United States throughout the area in which malaria occurred. However, any or all the members of the complex may have been regionally important, particularly in areas of most intense transmission.

Eco-ethological heterogeneity of the members of the Anopheles minimus complex (Diptera: Culicidae) in Southeast Asia and its consequences for vector control.

The presence of cryptic species within Anopheles minimus s.l. Theobald, one of the most widespread malaria vectors in Southeast Asia, was suggested on the basis of behavioral heterogeneities observed within this taxon. Subsequently, two species, A and C, were recognized. However, the existence of these cryptic species did not explain all observed behavioral heterogeneities within this complex. Besides, data on the behavior of vectors are essential to understand the dynamics of disease transmission and thus evaluate the appropriateness of vector control measures. Different collection methods were used to collect Anopheles species from several localities in Southeast Asia to assess the inter- and intraspecific behavioral divergences of An. minimus A and C. Collection results were subjected to a correspondence analysis. The members of the An. minimus complex were identified by use of the octanol dehydrogenase allozyme profiles or the polymerase chain reaction-restriction fragment length polymorphism assay. Large intraspecific behavioral differences were observed among populations of An. minimus A. These populations belong to the same species on the basis of the applied genetic markers. In northern Vietnam, species A tended to be more zoophilic, whereas in the study sites of south central Vietnam, Cambodia, and Laos it showed marked antropophilic behavior when cattle were scarce. In the most northern study site, An. minimus A showed noteworthy endophilic behavior. An. minimus C was primarily zoophilic and based on this behavior, its role in malaria transmission is questionable. However, it was only found in one locality, so that intraspecific behavioral variation could not be assessed. An. minimus A is able to change its host preference in function of local situations in host availability. Hence, its role in malaria transmission can differ from region to region. Similarly, the impact of vector control on this species may differ between localities. In conclusion, intraspecific behavioral differences in Anopheles species can occur and these behavioral heterogeneities, albeit important for disease transmission and control, are not a priori indicative for the presence of cryptic species.

Malaria transmission and major malaria vectors in different geographical areas of Southeast Asia.

During the last decade, major progress in malaria control has been achieved in Vietnam, Laos and Cambodia. However, malaria is still a potentially fatal disease in some hilly-forested areas and continues to be endemic in a few coastal foci. To estimate the risk that stems from the major vectors after a decade of intensive malaria control, an entomological study based on human landing collections was conducted between April 1998 and November 2000 in six study villages (four in Vietnam, one in Cambodia and one in Laos) located in different physio-geographical areas. Five villages were selected in places where new cases of malaria still occurred. In the sixth village, in the northern hilly area of Vietnam, no case of malaria was detected during the past 3 years. In three study villages of the hilly forested areas of Cambodia and central Vietnam, Anopheles dirus A still played an important role in malaria transmission and maintain perennial transmission inside the villages despite its low density. Anopheles minimus A was found in all study villages except in the southern coastal village of Vietnam. Its role in malaria transmission, however, varied between localities and surveys. In one study village of central Vietnam it was almost absent (one specimen collected over 480 man nights), and in another village sporozoite positive specimens (2.8%) were only observed during the first two surveys whereas this species disappeared from the collections from November 1998 onwards (six surveys: 360 man nights). In the northern study site An. minimus A and C were found in all collections, but no local malaria transmission occurred. However, the constant presence of these two species associated with a high longevity (parous rate up around 80% and 65%, respectively), suggests that transmission can occur at almost any time if parasite reservoirs are reintroduced in the area. The proper management of malaria cases and population movement is, therefore, important to prevent outbreaks and the reintroduction of malaria in northern Vietnam. In the study site of the Mekong delta, An. sundaicus occurred at high densities (up to 190 bites/man/night). The recent changes in land use from rice cultivation to shrimp farming probably explains the increase of this brackish water breeding species during the study period. However, none of the 11,002 specimens was positive for Plasmodium circumsporozoite protein (CSP). The relative low survival rate as estimated by the parous rate (around 47%) may reflect its low vectorial status that could explain the very low malaria incidence (1.9 case/100 persons/year) in this study site. A calculated sporozoite rate of maximum 1/300,000 is enough to explain this low malaria incidence. Despite the successes in malaria control, the vector An. dirus A continues to play an important role in malaria transmission, whereas An. minimus A showed temporal and spatial variation in its role as vector. The role of An. sundaicus as vector could not be confirmed because of the low incidence in the coastal study village. Other Anopheles species may be locally involved, but in the five study villages where malaria is still present they probably do not contribute significantly to malaria transmission. The study also points towards the fact that in Southeast Asia it will become increasingly difficult to incriminate Anopheles species in malaria transmission while the risk for malaria transmission still persist.

Population Structure of Anopheles gambiae in Africa.

The population structure of Anopheles gambiae in Africa was studied using 11 microsatellite loci in 16 samples from 10 countries. All loci are located outside polymorphic inversions. Heterogeneity among loci was detected and two putative outlier loci were removed from analyses aimed at capturing genome-wide patterns. Two main divisions of the gene pool were separated by high differentiation (F(ST) > 0.1). The northwestern (NW) division included populations from Senegal, Ghana, Nigeria, Cameroon, Gabon, Democratic Republic of Congo (DRC), and western Kenya. The southeastern (SE) division included populations from eastern Kenya, Tanzania, Malawi, and Zambia. Inhospitable environments for A. gambiae along the Rift Valley partly separate these divisions. Reduced genetic diversity in the SE division and results of an analysis based on private alleles support the hypothesis that a recent bottleneck, followed by colonization from the NW populations shaped this structure. In the NW division, populations possessing the M rDNA genotype appeared to form a monophyletic clade. Although genetic distance increased with geographic distance, discontinuities were suggested between certain sets of populations. The absence of heterozygotes between sympatric M and S populations in the DRC and the high differentiation in locus 678 (F(ST)>0.28) contrasted with low differentiation in all other loci (-0.02<F(ST)<0.09) and with the persistence of departures from Hardy-Weinberg expectations within each form in the DRC. Neither recent reproductive isolation alone nor selection alone can explain these results, a situation that is compatible with incipient speciation. Because it is possible that the molecular forms play different roles in malaria transmission, future studies should treat them separately.

Estimation of malaria transmission from humans to mosquitoes in two neighbouring villages in south Cameroon: evaluation and comparison of several indices

Malaria transmission from humans to mosquitoes was assessed in two neighbouring villages in a rural area near Yaoundé, Cameroon during high and low transmission seasons during 1998–2000, using several indices previously evaluated in different areas endemic for malaria but never directly compared. These indices were estimated from human parasitological data and mosquito infection rates and, for each individual, thick blood films were prepared at the same time as experimental infection of laboratory-bred mosquitoes. Among the 685 volunteers examined, the prevalence of Plasmodium falciparum gametocyte carriers was 16%, and 8% of individuals were able to infect mosquitoes. The percentage of mosquitoes that became infected by feeding on the infectious individuals was 21 %. Children aged < 10 years contributed to about 75% of the infectious reservoir, although they constituted only 35% of the total population. Differences were found between the transmission seasons and the villages, and varied according to the index examined. Although there were more infectious individuals in one of the two villages, they were less infectious than those in the other village during the high transmission season. Comparative analysis of the transmission indices suggests the existence of functioning transmission-blocking immunity in one of the villages, which until now has been only hypothetically considered to play a role in malaria transmission in a natural setting. The epidemiological value of all the indices used and their accuracy in estimating the human infectious reservoir and its natural or induced variations are discussed.