An evolutionary view of the interactions between anopheline mosquitoes and malaria parasites

The transmission of malaria is governed by the mosquito vector's biting rate, its mortality, and the developmental period of the parasite within the mosquito. This review covers some data on the interactions among these parameters and describes possible evolutionary mechanisms underlying two aspects of the parasite's life cycle.     

Host choice of anopheline mosquitoes in a malaria endemic area of western Venezuela

Abstract. Bloodmeals of exophilic anopheline mosquitoes collected resting on vegetation in a malaria endemic area in western Venezuela were identified by ELISA. Using a TMB peroxidase substrate in the ELISA, human bloodmeals were readily identified up to 40 h after ingestion in all laboratory-fed mosquitoes tested. Assay sensitivity declined to 75% identifiable 44 h post-feeding.
The Human Blood Index and the Feeding Index of each species differed between the three villages studied. An.triannulatus was generally more anthropophilic than An.nuneztovari and An.oswaldoi. These contrasting results emphasize the difficulties of interpreting host choice data.     

Illustrated keys to the anopheline mosquitoes of Myanmar.

In the newly revised and illustrated keys to 4th instar larvae and adult female mosquitoes, the following 36 Anopheles species from Myanmar are included: Anopheles aconitus, An. aitkenii, An. annularis, An. argyropus, An. barbirostris, An. bengalensis, An. culicifacies, An. dirus, An. fluviatilis, An. gigas, An. insulaeflorum, An. jamesii, An. jeyporensis, An. karwari, An. kochi, An. kyondawensis, An. lindesayi, An. maculatus, An. majidi, An. minimus, An. nigerrimus, An. nitidus, An. pallidus, An. peditaeniatus, An. philippinensis, An. pseudojamesii, An. sinensis, An. splendidus, An. stephensi, An. subpictus, An. sundaicus, An. tessellatus, An. theobaldi, An. vagus, An. varuna, and An. willmori. The new keys presented in this paper will enable public health workers to rapidly identify mosquito vectors of malaria and to distinguish them from other species in the same genera.     

Age-related effects on malaria parasite infection in wild chimpanzees

Wild great apes are widely infected with a number of malaria parasites (Plasmodium spp.). Yet, nothing is known about the biology of these infections in the wild. Using faecal samples collected from wild chimpanzees, we investigated the effect of age on Plasmodium spp. detection rates. The data show a strong association between age and malaria parasite positivity, with significantly lower detection rates in adults. This suggests that, as in humans, individuals reaching adulthood have mounted an effective protective immunity against malaria parasites.     

Adaptation of a strain of Plasmodium vivax from India to New World monkeys, chimpanzees, and anopheline mosquitoes.

A strain of Plasmodium vivax from India was adapted to develop in splenectomized Saimiri boliviensis, Aotus lemurinus griseimembra, A vociferans, A. nancymai, A. azarae boliviensis, hybrid Aotus monkeys, and splenectomized chimpanzees. Infections were induced via the inoculation of sporozoites dissected from the salivary glands of Anopheles stephensi and An. dirus mosquitoes to 12 Aotus and 8 Saimiri monkeys; transmission via the bites of infected An. stephensi was made to 1 Aotus monkey and 1 chimpanzee. The intravenous passage of infected erythrocytes was made to 9 Aotus monkeys and 4 chimpanzees. Gametocytes in 13 Aotus monkeys and 4 chimpanzees were infectious to mosquitoes. Infection rates were markedly higher in mosquitoes fed on chimpanzees. PCR studies on 10 monkeys injected with sporozoites revealed the presence of parasites before their detection by microscopic examination. The India VII strain of P. vivax develops in Aotus and Saimiri monkeys and chimpanzees following the injection of parasitized erythrocytes, or sporozoites, or both. The transmission rate via sporozoites to New World monkeys of approximately 50% may be too low for the testing of sporozoite vaccines or drugs directed against the exoerythrocytic stages. However, the strain is highly infectious to commonly available laboratory-maintained anopheline mosquitoes. Mosquito infection is especially high when feedings are made with gametocytes from splenectomized chimpanzees.     

Non-coding RNA gene families in the genomes of anopheline mosquitoes

Background

Only a small fraction of the mosquito species of the genus Anopheles are able to transmit malaria, one of the biggest killer diseases of poverty, which is mostly prevalent in the tropics. This diversity has genetic, yet unknown, causes. In a further attempt to contribute to the elucidation of these variances, the international “Anopheles Genomes Cluster Consortium” project (a.k.a. “16 Anopheles genomes project”) was established, aiming at a comprehensive genomic analysis of several anopheline species, most of which are malaria vectors. In the frame of the international consortium carrying out this project our team studied the genes encoding families of non-coding RNAs (ncRNAs), concentrating on four classes: microRNA (miRNA), ribosomal RNA (rRNA), small nuclear RNA (snRNA), and in particular small nucleolar RNA (snoRNA) and, finally, transfer RNA (tRNA).

Results

Our analysis was carried out using, exclusively, computational approaches, and evaluating both the primary NGS reads as well as the respective genome assemblies produced by the consortium and stored in VectorBase; moreover, the results of RNAseq surveys in cases in which these were available and meaningful were also accessed in order to obtain supplementary data, as were “pre-genomic era” sequence data stored in nucleic acid databases. The investigation included the identification and analysis, in most species studied, of ncRNA genes belonging to several families, as well as the analysis of the evolutionary relations of some of those genes in cross-comparisons to other members of the genus Anopheles.

Conclusions

Our study led to the identification of members of these gene families in the majority of twenty different anopheline taxa. A set of tools for the study of the evolution and molecular biology of important disease vectors has, thus, been obtained.

Electronic supplementary material

The online version of this article (doi:10.1186/1471-2164-15-1038) contains supplementary material, which is available to authorized users.     

Effects of salinity on the larvae of some Afrotropical anopheline mosquitoes

ABSTRACT. Laboratory experiments showed that the larvae of some freshwater-breeding Afrotropical anopheline species exhibit various degrees of tolerance to 25% sea-water. At 24^oC, survival rates from egg to fourth instar larva were significantly less in salt-water than in freshwater for four species: 39.5% versus 55.4% for Anopheles tenebrosus Dönitz, 31.9% versus 86.2% for An. mousinhoi De Meillon & Pereira, 3.5% versus 67.9% for An. pharoensis Theobald and 3.9% versus 41.9% for An. quadriannulatus (Theobald). Conversely, An. merus Dönitz showed a significantly better survival rate of 46.4% in 25% sea-water compared with 15.5% in freshwater. Simulated winter conditions did not significantly change these differential survival rates of An. merus. Effects of salinity on the anal papillae of third instar larvae are discussed.     

Malaria infection potential of anopheline mosquitoes sampled by light trapping indoors in coastal Tanzanian villages

Abstract. Anopheline mosquito populations were studied during 1992 in seven villages south of Bagamoyo, coastal Tanzania, prior to malaria control intervention using insecticide treated bednets. To collect mosquitoes, CDC light traps were used in ten houses per village fortnightly for 12 months. Anopheles females were identified and checked by ELISA for the presence of malaria sporozoite antigen and source of bloodmeal. An. funestus peaked in June-July after the long rains. Three members of the An. gambiae complex had different seasonality: An. arabiensis, An. gambiae and small numbers of An. merus were collected.
In most villages transmission was extremely high and perennial with the entomological inoculation rate reaching three to eleven infective bites per person per night in July and persisting at around 0.1 and 1 for most of the remainder of the year. Sporozoite infection rates within the An. gambiae complex ranged from 2% to 25%, with the peaks in January and July following the two rainy periods. An. funestus showed a similar pattern. The light traps were reliable, simple to operate, and proved to be satisfactory to study the mosquito vector population.     

Seasonal abundance of anopheline mosquitoes and their association with rainfall and malaria along the Matapí River,Amapí, Brazil

Three communities separated by 1.5–7.0 km, along the Matapí River, Amapá State, Brazil, were sampled monthly from April 2003 to November 2005 to determine relationships between seasonal abundance of host‐seeking anophelines, rainfall and malaria cases. Out of the 759 821 adult female anophelines collected, Anopheles darlingi Root (Diptera: Culicidae) was the most abundant (56.2%) followed by An. marajoara Galvão & Damasceno (24.6%), An. nuneztovari Gabaldón (12.4%), An. intermedius (Chagas) (4.4%) and An. triannulatus (Neiva and Pinto) (2.3%). Vector abundance, as measured by human landing catches, fluctuated during the course of the study and varied in species‐specific ways with seasonal patterns of rainfall. Anopheles darlingi and An. triannulatus were more abundant during the wet‐dry transition period in June to August, whereas An. marajoara began to increase in abundance in February in two villages, and during the wet‐dry transition in the other village. Anopheles nuneztovari and An. intermedius increased in abundance shortly after the rains began in January to February. A generalized linear mixed model (GLMM) analysis of 32 consecutive months of collections showed significant differences in abundance for each species by village and date (P < 0.0001). Correlations between lagged rainfall and abundances also differed among species. A strong positive correlation of An. darlingi abundance with rainfall lagged by 4 and 5 months (Pearson's r = 0.472–0.676) was consistent among villages and suggests that rainfall may predict vector abundance. Significant correlations were detected between numbers of malaria cases and abundances of suspected vector species. The present study shows how long‐term field research may connect entomological and climatological correlates with malaria incidence.     

Comparison between anopheline mosquitoes (Diptera: Culicidae) caught using different methods in a malaria endemic area of Papua New Guinea

The mosquito sampling efficiency of CDC (Centers for Disease Control) miniature light traps hung adjacent to mosquito nets, was compared with that of both indoor and outdoor human-bait collections in ten villages in the Wosera area of Papua New Guinea. The most frequently collected anopheline in the matched indoor and light trap samples was Anopheles koliensis Owen, followed by A. punctulatus D?nitz, A. karwari (James), A. farauti Laveran (sensu lato), A. longirostris Brug and A. bancroftii Giles. All species were much less frequent in the light traps than in landing catches. The hypothesis that the numbers of mosquitoes in light traps are proportional to human landing catches was examined using regression models that allowed for sampling error in both entomological measurements. Light traps under-sampled A. punctulatus and A. farauti s.l. at high densities. The models indicated that the ratio of light trap to landing catch females of A. koliensis and A. karwari increased with increasing mosquito density. Light trap catches of A. longirostris were proportional to indoor landing rates but when outdoor landing rates were high this species was under-sampled by light traps. Numbers of A. bancroftii in light traps were found to be proportional to those in outdoor landing catches, but were negatively related to those attempting to bite indoors. Circumsporozoite positivity rates for both Plasmodium falciparum Welch and P. vivax (Grassi & Feletti) in A. punctulatus and A. farauti s.l. were significantly higher in light trap collections than in either indoor or outdoor landing catches, suggesting that light traps may selectively sample older mosquitoes of these species.     

A theoretical approach to predicting the success of genetic manipulation of malaria mosquitoes in malaria control

Background  

Mosquitoes that have been genetically modified to better encapsulate the malaria parasite Plasmodium falciparum are being considered as a possible tool in the control of malaria. Hopes for this have been raised with the identification of genes involved in the encapsulation response and with advances in the tools required to transform mosquitoes. However, we have only very little understanding of the conditions that would allow such genes to spread in natural populations.     

Analysis of the sporogonic development of Plasmodium falciparum and Plasmodium berghei in anopheline mosquitoes

Basic knowledge of the sporogonic development of malarial parasites is crucial when evaluating the sporontocidal activity of antimalarial drugs or when determining why certain vectors are refractory to a particular parasite while others are competent vectors. We have developed a model which we have used to i) assess the sporogonic development of Plasmodium berghei ANKA in Anopheles stephensi and A. freeborni mosquitoes and ii) determine the effect of chloroquine on the sporogony of P. falciparum NF-54 in A. stephensi. Criteria used to assay sporogonic development include: i) number of oocysts present, ii) percentage of mosquitoes with oocysts, iii) time of release of sporozoites from the oocysts into the hemolymph, iv) time and degree of sporozoite invasion of salivary glands, and v) transmission (P. berghei) into vertebrate hosts. Parasite development in the mosquito is evaluated every other day, commencing on ca. day 7 post-feed (PF) and continuing until ca. day 22 PF. These detailed observations allow us to delineate the chronology of sporogonic development.     

From nest to nest--influence of ecology and reproduction on the active period of adult Gombe chimpanzees

The time spent between sleeping periods, which is called the active period, has to accommodate all essential activities, including feeding, resting, social behavior, and reproduction. To minimize costs in terms of, e.g., predation risk, suboptimal foraging, or sleep deficiency, the active period of diurnal animals should be less than or equal to the daylight period. Thus, the active period of an animal should be shaped by local environmental conditions as well as by metabolic and reproductive demands. Chimpanzees, which exhibit reduced predator pressure and a flexible fission-fusion society, were chosen as a model to explore these links. We investigated the influence of sex, female reproductive status, dominance rank, and season on the duration of the active period of adult chimpanzees at Gombe National Park, Tanzania (1975-1992). Sexually nonreceptive females had shorter active periods compared to males, while receptive females had even longer active periods than males. Dominance rank did not influence the duration of the active period of nonreceptive females, but high- and middle-ranking males had shorter active periods compared to low-ranking males. Nonreceptive females exhibited longer active periods during the dry season than in the wet season. No seasonal effect was discovered for males, perhaps because they already had long active periods in the wet season. Nonreceptive females seem to be able to accommodate all essential activities in the daylight period available, probably because they live less socially than males. Thus, the active period does not reflect differences in female competitive abilities, but does reflect such differences in males. The duration of the active period appears to be a simple, reliable tool for exploring basic responses and constraints in animal societies.     

Molecular surveillance of malaria in the Central Highlands,Vietnam

Vietnam achieved outstanding success against malaria in the last few decades. The mortality and morbidity of malaria in Vietnam have decreased remarkably in recent years, but malaria is still a major public health concern in the country, particularly in the Central Highlands region. In this study, molecular analyses of malaria parasites in the Central Highlands were performed to understand the population structure and genetic diversity of the parasites circulating in the region. Plasmodium falciparum (68.7%) and P. vivax (27.4%) along with mixed infections with P. falciparum/P. vivax (3.9%) were detected in 230 blood samples from patients with malaria. Allele-specific nested-polymerase chain reaction (PCR) or PCR-restriction fragment length polymorphism (PCR-RFLP) analyses of pfmsp-1, pfama-1, pvcsp, and pvmsp-1 revealed complex genetic makeup in P. falciparum and P. vivax populations of Vietnam. Substantial multiplicity of infection (MOI) was also identified, suggesting significant genetic diversity and polymorphism of P. falciparum and P. vivax populations in the Central Highlands of Vietnam. These results provide fundamental insight into the current patterns of dispersion and genetic nature of malaria parasites as well as for the development of malaria elimination strategies in the endemic region.     

The effects of age, exposure history and malaria infection on the susceptibility of Anopheles mosquitoes to low concentrations of pyrethroid

Chemical insecticides are critical components of malaria control programs. Their ability to eliminate huge numbers of mosquitoes allows them to swiftly interrupt disease transmission, but that lethality also imposes immense selection for insecticide resistance. Targeting control at the small portion of the mosquito population actually responsible for transmitting malaria parasites to humans would reduce selection for resistance, yet maintain effective malaria control. Here, we ask whether simply lowering the concentration of the active ingredient in insecticide formulations could preferentially kill mosquitoes infected with malaria and/or those that are potentially infectious, namely, old mosquitoes. Using modified WHO resistance-monitoring assays, we exposed uninfected Anopheles stephensi females to low concentrations of the pyrethroid permethrin at days 4, 8, 12, and 16 days post-emergence and monitored survival for at least 30 days to evaluate the immediate and long-term effects of repeated exposure as mosquitoes aged. We also exposed Plasmodium chabaudi- and P. yoelii-infected An. stephensi females. Permethrin exposure did not consistently increase mosquito susceptibility to subsequent insecticide exposure, though older mosquitoes were more susceptible. A blood meal slightly improved survival after insecticide exposure; malaria infection did not detectably increase insecticide susceptibility. Exposure to low concentrations over successive feeding cycles substantially altered cohort age-structure. Our data suggest the possibility that, where high insecticide coverage can be achieved, low concentration formulations have the capacity to reduce disease transmission without the massive selection for resistance imposed by current practice.     

Distribution of African malaria mosquitoes belonging to the Anopheles gambiae complex

The distribution of malaria vector mosquitoes, especially those belonging to species complexes that contain non-vector species, is important for strategic planning of malaria control programmes. Geographical information systems have allowed researchers to visualize distribution data on maps together with environmental parameters, such as rainfall and temperature. Here, Maureen Coetzee, Marlies Craig and David le Sueur review our current knowledge on the distribution of the members of the Anopheles gambiae complex.     

Seasonal population density and daily survival of anopheline mosquitoes (Diptera: Culicidae) in a malaria endemic area, Republic of Korea.

Mosquito surveillance was conducted near the Korean Demilitarized Zone (Paju County, Gyeonggi Province) from April to October, 1999, where malaria cases were reported. Adult mosquito surveillance, using black light and CDC UV light traps, was conducted at five and two sites, respectively. Weekly larval collections were made at five rice paddies located adjacent to the adult collection sites. Anopheles sinensis was the most abundant mosquito of 11 species collected throughout the surveillance period in 1999, comprising 47 - 48% of the total number of mosquitoes collected at cow sheds and residence. At all five sites surveyed by CDC UV light traps, anophelines appeared early in the year (May 3) and were most abundant in the cow sheds followed by the hillside forest, residence, stream/river bank, and were least abundant in rice fields. The population density of the larvae and the adults of An. sinensis increased steadily in June and reached their peaks during the second week of July (mean 112 females/trap/night). The parity rates were higher in July and September, when populations were highest. The probabilities of daily survival of An. sinensis were 0.804 in June to 0.895 in July. Cross-correlation showed a significant relationship between the number of adult anopheline mosquitoes and the number of larvae collected on the previous day, the same day, and also three and seven days later, which may be useful for determining treatment thresholds.     

Wolbachia strain wAlbB enhances infection by the rodent malaria parasite Plasmodium berghei in Anopheles gambiae mosquitoes

Wolbachia, a common bacterial endosymbiont of insects, has been shown to protect its hosts against a wide range of pathogens. However, not all strains exert a protective effect on their host. Here we assess the effects of two divergent Wolbachia strains, wAlbB from Aedes albopictus and wMelPop from Drosophila melanogaster, on the vector competence of Anopheles gambiae challenged with Plasmodium berghei. We show that the wAlbB strain significantly increases P. berghei oocyst levels in the mosquito midgut while wMelPop modestly suppresses oocyst levels. The wAlbB strain is avirulent to mosquitoes while wMelPop is moderately virulent to mosquitoes pre-blood meal and highly virulent after mosquitoes have fed on mice. These various effects on P. berghei levels suggest that Wolbachia strains differ in their interactions with the host and/or pathogen, and these differences could be used to dissect the molecular mechanisms that cause interference of pathogen development in mosquitoes.