Pyrethroid resistance in Anopheles gambiae s.s. and Anopheles arabiensis in western Kenya: phenotypic,metabolic and target site characterizations of three populations

Field and laboratory investigations revealed phenotypic, target site and metabolic resistance to permethrin in an Anopheles gambiae s.s. (Diptera: Culicidae) population in Bungoma District, a region in western Kenya in which malaria is endemic and rates of ownership of insecticide‐treated bednets are high. The sensitivity of individual An. gambiae s.l. females as indicated in assays using World Health Organization (WHO) test kits demonstrated reduced mortality in response to permethrin, deltamethrin and bendiocarb. Estimated time to knock‐down of 50% (KDT₅₀) of the test population in Centers for Disease Control (CDC) bottle bioassays was significantly lengthened for the three insecticides compared with that in a susceptible control strain. Anopheles arabiensis from all three sites showed higher mortality to all three insecticides in the WHO susceptibility assays compared with the CDC bottle assays, in which they showed less sensitivity and longer KDT₅₀ than the reference strain for permethrin and deltamethrin. Microplate assays revealed elevated activity of β‐esterases and oxidases, but not glutathione‐S‐transferase, in An. gambiae s.s. survivors exposed to permethrin in bottle bioassays compared with knocked down and unexposed individuals. No An. arabiensis showed elevated enzyme activity. The 1014S kdr allele was fixed in the Bungoma An. gambiae s.s. population and absent from An. arabiensis, whereas the 1014F kdr allele was absent from all samples of both species. Insecticide resistance could compromise vector control in Bungoma and could spread to other areas as coverage with longlasting insecticide‐treated bednets increases.     

Conspecific Sharing of Breeding Sites by Anopheline Female Mosquitoes (Diptera: Culicidae) Inferred from Microsatellite Markers

The number of Anopheles gambiae and Anopheles arabiensis females that used each of the 33 sampled breeding sites in west Kenya was estimated by microsatellite markers and related statistics to test the hypothesis that conspecific females share aquatic sites. Totally, 166 An. gambiae and 168 An. arabiensis larvae were identified and were genotyped. The mean number of larvae per breeding site was 8.3 for An. gambiae and 8.4 for An. arabiensis. The likelihood method estimated that, for An. gambiae, the mean number of females that would have laid eggs per breeding site was 5.2 and ranged from 2 to 9, and for An. arabiensis, the mean was 5.0 with a range of 2–10. The clustering method estimated that the mean number of females laying eggs per breeding site was 6.8 for An. gambiae. The results provide molecular evidence that females of one or both species share breeding sites.     

Insecticide resistance monitoring of field‐collected Anopheles gambiae s.l. populations from Jinja,eastern Uganda,identifies high levels of pyrethroid resistance

Insecticide resistance in the malaria vector Anopheles gambiae s.l. (Diptera: Culicidae) threatens insecticide‐based control efforts, necessitating regular monitoring. We assessed resistance in field‐collected An. gambiae s.l. from Jinja, Uganda using World Health Organization (WHO) biosassays. Only An. gambiae s.s. and An. arabiensis (?70%) were present. Female An. gambiae exhibited extremely high pyrethroid resistance (permethrin LT₅₀ > 2 h; deltamethrin LT₅₀ > 5 h). Female An. arabiensis were resistant to permethrin and exhibited reduced susceptibility to deltamethrin. However, while An. gambiae were DDT resistant, An. arabiensis were fully susceptible. Both species were fully susceptible to bendiocarb and fenitrothion. Kdr 1014S has increased rapidly in the Jinja population of An. gambiae s.s. and now approaches fixation (?95%), consistent with insecticide‐mediated selection, but is currently at a low frequency in An. arabiensis (0.07%). Kdr 1014F was also at a low frequency in An. gambiae. These frequencies preclude adequately‐powered tests for an association with phenotypic resistance. PBO synergist bioassays resulted in near complete recovery of pyrethroid susceptibility suggesting involvement of CYP450s in resistance. A small number (0.22%) of An. gambiae s.s. ×An. arabiensis hybrids were found, suggesting the possibility of introgression of resistance alleles between species. The high levels of pyrethroid resistance encountered in Jinja threaten to reduce the efficacy of vector control programmes which rely on pyrethroid‐impregnated bednets or indoor spraying of pyrethroids.     

DNA probes for species identification of mosquitoes in the Anopheles gambiae complex

Abstract. Identification of species within the Anopheles gambiae Giles species complex is essential for the correct evaluation of malaria vector ecology studies and control programmes. The development of DNA probes to distinguish species of the An.gambiae complex is described. Genomic libraries were prepared for four members of the An.gambiae complex. These were screened using radiolabeled DNA from different species of An. gambiae sensu lato and a number of clones selected on the basis of their species specificity. These clones could be divided into two groups, each containing homologous sequences. Sequences homologous to group 1 inserts are highly reiterated in the genomes of Anopheles arabiensis Patton and Anopheles merus Dönitz, present in low copy number in Anopheles melas Theobald, but were not detected in Anopheles gambiae sensu stricto. Studies on the organization of this sequence in the genome of An.arabiensis show that homologous sequences are male specific and interspersed within the chromatin. Sequences homologous to group 2 inserts are highly repeated in the genomes of An.merus and An.melas, but present in low copy number in An.gambiae s.s. and An.arabiensis. Group 2 homologous sequences are not sex-specific in the species tested and appear to be tandemly repeated. When used as hybridization probes, these sequences provide a sensitive means for the identification of species within the Anopheles gambiae complex.     

Application of a reverse dot blot DNA–DNA hydridization method to quantify host‐feeding tendencies of two sibling species in the Anopheles gambiae complex

A DNA–DNA hybridization method, reverse dot blot analysis (RDBA), was used to identify Anopheles gambiae s.s. and Anopheles arabiensis (Diptera: Culicidae) hosts. Of 299 blood‐fed and semi‐gravid An. gambiae s.l. collected from Kisian, Kenya, 244 individuals were identifiable to species; of these, 69.5% were An. arabiensis and 29.5% were An. gambiae s.s. Host identifications with RDBA were comparable with those of conventional polymerase chain reaction (PCR) followed by direct sequencing of amplicons of the vertebrate mitochondrial cytochrome b gene. Of the 174 amplicon‐producing samples used to compare these two methods, 147 were identifiable by direct sequencing and 139 of these were identifiable by RDBA. Anopheles arabiensis bloodmeals were mostly (94.6%) bovine in origin, whereas An. gambiae s.s. fed upon humans more than 91.8% of the time. Tests by RDBA detected that two of 112 An. arabiensis contained blood from more than one host species, whereas PCR and direct sequencing did not. Recent use of insecticide‐treated bednets in Kisian is likely to have caused the shift in the dominant vector species from An. gambiae s.s. to An. arabiensis. Reverse dot blot analysis provides an opportunity to study changes in host‐feeding by members of the An. gambiae complex in response to the broadening distribution of vector control measures targeting host‐selection behaviours.     

The association between the phytoplankton, Rhopalosolen species (Chlorophyta; Chlorophyceae), and Anopheles gambiae sensu lato (Diptera: Culicidae) larval abundance in western Kenya

Algae are important food resources of the larvae of the African malaria vectors, Anopheles gambiae Giles and Anopheles arabiensis Patton (Anopheles gambiae sensu lato), and other zooplankton, but empirical evidence remains meager about the agal flora in ephemeral water bodies. The animals present in natural aquatic habitats in western Kenya were sampled from July to November 2002 to study abiotic and biotic environmental factors determining A. gambiae sl larval abundance. The five highest concentrations of third and fourth instars and pupae (hereafter referred to as old-stage larvae) were sampled in conjunction with the unicellular epizoic green algae, Rhopalosolen species (Chlorophyta; Chlorophyceae). Canonical correspondence analysis revealed that the presence of Rhopalosolen species was the most important determinant of the animal assemblage. The density of old-stage A. gambiae sl larvae was positively correlated with the presence of Rhopalosolen species, but the density of first and second instars of A. gambiae sl was not. The water bodies with Rhopalosolen sp. yielded larger mosquitoes in spite of the higher density of larvae. We demonstrated that the productivity of water bodies in terms of the larvae of malaria vectors can differ in magnitude depending on the agal flora. We discuss phytoplankton as a regulator of mosquito larval populations.     

A survival and reproduction trade-off is resolved in accordance with resource availability by virgin female mosquitoes

The first 2-4 days after an Anopheles gambiae female mosquito emerges are critical to her survival and reproductive success. Yet, the order of behavioural events (mating, sugar feeding, blood feeding) during this time has received little attention. We discovered that among female cohorts sampled from emergence, sugar feeding had a higher probability than blood feeding of occurring first, and mating rarely occurred before a meal was taken. The night after emergence, 48% of females fed on sugar in mesocosms, and 25% fed on human blood; in the absence of sugar, 49% of females fed on human blood. After 5 days, 39% of the sugar-supplied females had blood fed and mated, and were fructose negative, whereas only 8% of the sugar-denied females had both blood fed and mated by this time. The model that best explained the transitions suggests that females made use of two distinct behavioural pathways, the most common one being to sugar-feed, then mate, and then seek blood. Other females sought blood first, then mated, and forwent a sugar meal. Lipid levels were higher in females with access to sugar than in females without access to sugar, particularly for those in later gonotrophic stages, while glycogen levels in the sugar-supplied group were higher throughout. In single-night experiments with females having had access to sucrose since emergence, those given a blood meal 1 day before spending a night with males had higher insemination rates than those not receiving the blood meal. These results indicate that the trade-off between survival and immediate reproduction is resolved by young adult females in accordance with availability of resources and gonotrophic state.     

Use of rDNA-PCR to investigate the ecological distribution of Anopheles bwambae in relation to other members of the An.gambiae complex of mosquitoes in Bwamba County, Uganda

Abstract Environmental relationships were investigated among three species of the Anopheles gambiae complex of mosquitoes associated with the geothermal springs located in Bwamba County, Uganda. The degree of ecological isolation between An.gambiae and An. bwambae, a sibling species known only from the geothermal springs environment, was assessed on the basis of adult distribution and abundance as well as differences in larval habitats. Field data were gathered during June 1995 without knowing which of the species were being collected. Specimens identified subsequently by rDNA-PCR were used to interpret the ecological data. Ten of twenty aquatic sites sampled were found positive for immature stages of the An.gambiae complex. Larvae of An.bwambae were associated with ‘springwater’ habitats having much higher conductivity, much greater concentrations of dissolved solids and slightly higher temperature and pH than ‘normal’ fresh water sites inhabited by larvae of An.gambiae. Larval habitats of both species were unshaded: An.bwambae occurred among dense sedge (Cyperus laevigatus) whereas those of An.gambiae were almost devoid of vegetation. One mixed sample showed that larvae of both species occur together in peripheral aquatic sites with intermediate physical and ecological characteristics. In water preference tests, free-flying females were reluctant to lay eggs on bowls of water in cages; gravid females (with one wing amputated) placed on the surface of water in a cup laid eggs on seasoned rainwater (12/51 An.bwambae; 2/3 An.gambiae) as well as spring-water (39/51 An.bwambae; 1/3 An.gambiae). All three An.gambiae oviposited on the first water option, whereas 86% of An. bwambae witheld oviposition until being moved to the other type of water after 5–6 h, and 82% (36/44) of these laid eggs on geothermal water in preference to rainwater. Larval and adult collections showed that An.gambiae occurs sympatrically with An.bwambae throughout its range in the humid foothill environment of the geothermal springs, whereas the distribution of An.arabiensis overlaps only slightly with An.bwambae towards the savanna environment north of the springs.     

Tick infestation risk and <Emphasis Type="Italic">Borrelia burgdorferi</Emphasis> s.l. infection-induced increase in host-finding efficacy of female <Emphasis Type="Italic">Ixodes ricinus</Emphasis> under natural conditions

An investigation of the risk of human tick infestation, together with the prevalence of Borrelia burgdorferi s.l. infection, was conducted in a sylvatic habitat in western Germany to provide data needed for future risk-benefit evaluations of acaricides used for clothing impregnation. Additionally, data were collected on behavioural changes in Borrelia burgdorferi s.l.-infected adult female I. ricinus ticks and the possible impact of such changes on host-finding efficacy. The risk of I. ricinus-infestation was determined by collecting from the protective clothing of volunteers and by dragging in known tick-infested sites in the Kühkopf Mountain area, Koblenz, Germany, from June through October 2006. The overall tick infestation rate per person per hour was 7.4 ± 5.5, with the following sex- and stage-specific differences: males 0.32 ± 0.37, females 1.1 ± 1.2, nymphs 3.6 ± 4.4, larvae 2.4 ± 3.5. Concurrent dragging revealed an average 19.4 ± 16.2 times higher infestation rate as well as a markedly lower infection rate with borreliae in adult I. ricinus ticks when compared to ticks collected from exposed human volunteers. Although the difference in infection rates was statistically significant (P < 0.023) only in adult female ticks, our data indicate that B. burgdorferi s.l. infection may increase host-finding efficacy in adult I. ricinus. The overall exposure risk was 1.0 B. burgdorferi s.l.-infected ticks per person per hour of exposure, or 0.25 ticks per 100 m walking distance in the study area.     

Protandry in the parasitoidCardiochiles nigriceps,as related to its mating system

A laboratory study was conducted of the emergence times and mating success ofCardiochiles nigriceps Viereck (Hymenoptera: Braconidae), a larval parasitoid of the tobacco budworm,Heliothis virescens (F.) (Lepidoptera: Noctuidae), and in view of this information, the parasitoid's mating system was explored. Observations on laboratory populations ofC. nigriceps confirmed the occurrence of two types of protandry, i.e. seasonal and diurnal; males emerged 2 days before females in a generation, and emerged about 1 hr before females on a given day. An experiment on mating success showed that newly emerged females are not receptive to males that emerged 1 hr earlier, but that these males and females often mate successfully after 1 more hr. The experiment also showed that 1- to 5-day-old males are more successful than 1-hr-old males in mating with 0- to 1-hr-old females. Thus, it is argued that males emerging on a given day have a disadvantage in competition for mates with males that emerged days earlier, and that this disadvantage may serve as a selection pressure toward diurnal protandry. A monogamous mating system for females ofC. nigriceps is suggested because sexual selection would be expected to be strong in species exhibiting both seasonal and diurnal protandry. A possibility of a sibling mating system inC. nigriceps is questioned partly because newly emerged females are unreceptive to males that emerged 1 hr earlier and partly because this parasitoid is solitary in what is considered highly dispersed hosts in the field.     

Studies on the critical water mass and the rehydration potential of unfed adult Dermacentor marginatus and D. reticulatus ticks (Acari: Ixodidae)

In unfed adult Dermacentor marginatus and D. reticulatusticks survival and capability to restore water balance after loss of high percentages of exchangeable body water were investigated. Furthermore, it was examined how frequently dehydrated ticks of these species were able to rehydrate by uptake of atmospheric water vapour. The critical water mass, defined as the water mass remaining in a tick at the nonambulatory state, differed between light and heavy weight groups and averaged 62.4 and 55.8%, respectively, of the total body water of fully hydrated ticks in females, and 54.4 and 51.1% respectively, in males of D. marginatus. In D. reticulatus, the corresponding figures were 55.9 and 54.7% in females and 52.1 and 52.7% in males. All ticks survived dehydration to 50, 75 or 100% of the critical water mass, and 96.7% of the D. marginatus ticks and 95.8% of the D. reticulatus ticks compensated water losses during subsequent incubation at 95% relative humidity (r.h.) and 20°C. Unfed females and males of both Dermacentor spp. were capable to balance water loss very frequently over a period of several months. When ticks were repeatedly dehydrated at 0% r.h. for 7 days and rehydrated at 95% r.h. and 20°C, females and males of D. marginatus reached the 50% mortality after 22 and 29 cycles of de- and rehydration, respectively, during 211 and 285 days, respectively. In D. reticulatus, 50% of females and males survived 23 and 17 cycles, respectively, during 248 and 186 days, respectively. Rehydration weights were as high or even higher as those of ticks kept at permanent 95% r.h.     

Factors influencing parasitism of adult Japanese beetles,Polillia japonica (Col.: Scarabaeidae) by entomopathogenic nematodes

Several factors that influence the activity of steinernematid and heterorhabditid nematodes against adult Japanese beetles were examined in the laboratory. The effect of nematode concentration on mortality of adult beetles was evaluated using a Petri plate bioassay. The adults were exposed to 1,000 to 10,000 infective stage juveniles (J3) ofSteinernema glaseri per 10 beetles with or without food for 24 hr after which they were held with food for an additional 6 days. The LC50s for males with and without food during exposure were 3,435 and 2,854 J3s/10 adults, respectively. The LC50s for mixtures of males and females with and without food were 5,228 and 1,762 J3s/10 adults respectively. Although mortality occurred during and shortly after exposure, significant additional mortality was observed 1–4 days following exposure. Exposure of males and females with food to 10,000 J3s/10 adults for 6, 12, 18 or 24 hr resulted in 47, 58, 72 and 77% mortality, respectively. Comparative activity ofS. glaseri, S. carpocapsae (All strain),S. feltiae (Biosys experimental cold adapted strain=bibionis),S. feltiae (Biosys experimental strain 27),Heterorhabditis bacteriophora, andHeterorhabditis sp. (Terceiran isolate) was evaluated against adult Japanese beetles using a 24 hr exposure to 8,000 J3s/10 adults. The most virulent species wereS. glaseri, S. feltiae (=bibionis), the Terceiran isolate ofHeterorhabditis andS. carpocapsae producing 55, 44, 36 and 34% mortality respectively. Our results indicate that adult Japanese beetles infected with entomopathogenic nematodes could serve as a mechanism for nematode dispersal.     

Odor-mediated flight behavior ofAnopheles gambiae gilesSensu Stricto andAn. stephensi liston in response to CO2, acetone, and 1-octen-3-ol (Diptera: Culicidae)

The flight behavior of Anopheles gambiae s.s. Giles and An. stephensi Patton exposed to different odor cues was studied in a wind tunnel. Odors consisted of CO₂, CO₂ + acetone (at two concentrations), and CO₂ + 1-octen-3-ol. Mosquitoes were released singly and their behavior was recorded on video. Parameters studied included flight velocity, percentage of time spent flying, percentage of time spent in plume, and number of turns toward the plume. Large differences in behavior toward the odors tested were observed. An. gambiaedid not respond well to CO ₂,whereas An. stephansiwas positively affected by this compound. In contrast, An. gambiaeresponded significantly to CO ₂ + acetone (at a low concentration), but the behavior of An. stephensiwas completely suppressed by this combination of odor stimuli. CO ₂ + a high concentration of acetone or CO ₂ + 1-octen-3-ol did not cause significant effects in An. gambiaecompared to no odor, while these treatments elicited strong behavioral responses in An. stephensi.The latter species responded particularly well to CO ₂ + 1-octen-3-ol. The results suggest that the observed differences may be inherent to the known differences in host preferences, where An. gambiaeis highly anthropophilic and An. stephensimore zoophilic. This would explain why the latter species responds well to CO ₂ and even better to CO ₂ + 1-octen-3-ol, a compound readily emitted by bovine ruminants.     

Biological differences in reproductive strategy between the mosquito sibling species Anopheles gambiae sensu stricto and An. quadriannulatus

Females of the afrotropical mosquito species Anopheles gambiae Giles sensu stricto and An. quadriannulatus (Theobald) (Diptera: Culicidae) were studied for the effect of blood meal size and the frequency of blood feeding on reproductive development during the first gonotrophic cycle. To standardize the blood meals, meals were administered by enema in some experiments. The effects of insemination, mosquito size, and metabolic reserves at emergence on egg development were also investigated. Maximum insemination was reached after seven days, varying from 62% in An. quadriannulatus to 95% in An. gambiae and was significantly different (P<0.05) between the two species. Insemination had no effect on feeding success. Females of An. quadriannulatus were significantly larger than An. gambiae females (mean wing size 2.90 ±0.01 mm versus 2.82 ±0.01 mm), but the protein, glycogen, and lipid content of newly emerged females of the two species were not significantly different. Without a blood meal, larger females of both species were significantly more likely to develop oocytes up to Christopher's stage II. With one blood meal, 27% of An. gambiae became pre-gravid and 73% matured eggs. In contrast, all An. quadriannulatus females remained in the pre-gravid stage following ingestion of one blood meal. Vitellogenesis was significantly reduced in smaller-sized pre-gravid An. quadriannulatus compared to larger individuals. When given the opportunity to feed up to three times on three successive days, all females of An. gambiae matured eggs but only 85% of An. quadriannulatus did so. When 1 l of human blood was administered by enema, none of the females of either species developed eggs. With a single enema of 1.5 l of human blood, only An. gambiae developed eggs. A similar result was observed with 1 l and 1.5 l enemas of bovine blood although some An. gambiae also developed eggs with 1 l of blood. Anopheles quadriannulatus developed eggs only when given two 1 l enemas on successive days. However, the percentage of females developing eggs was significantly lower than that of An. gambiae. The implications of these differences in reproductive strategy are discussed in the light of behavioural traits in the field.     

Longitudinal follow‐up of malaria transmission dynamics in two villages in a Sahelian area of Niger during a nationwide insecticide‐treated bednet distribution programme

Malaria transmission was monitored in two villages in the Sahel zone of Niger over 4 years. During this period, a nationwide vector control programme was carried out in which insecticide‐treated bednets were distributed free to mothers of children aged <5 years. Anopheles gambiae and Anopheles arabiensis (Diptera: Culicidae) were found to be the major malaria vectors. The dynamics of An. gambiae s.l. did not vary dramatically over the study period although the proportion of female mosquitoes found resting indoors decreased in both villages and, in one village, the parity rate and sporozoite index were significantly reduced after bednet distribution. By contrast with An. gambiae, the dynamics of Anopheles funestus altered greatly after the bednet distribution period, when adult density, endophagous rate and sporozoite rates decreased dramatically. Our observations highlight the importance of quantifying and monitoring the dynamics and infections of malaria vectors during large‐scale vector control interventions.     

Activity and mating competitiveness of γHCH/dieldrin resistant and susceptible male and virgin female Anopheles gambiae and An.stephensi mosquitoes, with assessment of an insecticide-rotation strategy

The effects of gamma HCH/dieldrin resistance genes on flight activity and mating competitiveness were investigated in males from backcrossed strains of Anopheles gambiae Giles and An.stephensi Liston. Activity of males and virgin females of both species, as recorded in an acoustic actograph, occurred mainly at dusk (the E peak). The activity pattern of An.gambiae males was not affected by resistance genes; in mating competition and predator avoidance experiments, however, RR males were less successful than RS males which were less successful than SS males. The activity pattern of An.stephensi differed from An.gambiae in that the E peaks of RR males and females in a gradual dusk regime were out of synchrony with those of SS and RS, the E peaks of RR occurring slightly later. Thus, RR males and females tended to mate assortatively in mate competition experiments. When a sudden dusk regime was substituted for the gradual dusk regime, activity of RR An.stephensi became synchronized with SS and RS activity, but in mating competition experiments RR still tended to mate assortatively. Estimates of male competitiveness, together with previously-obtained estimates of female fitness, were included in population genetics models. Computer simulations showed that the frequency of resistance in populations of An.gambiae and An.stephensi should decrease in the absence of insecticide at a rate comparable with known field reversions.     

Interspecific competition between sibling species larvae of Anopheles arabiensis and An. gambiae

Mosquito larvae of the sibling species Anopheles arabiensis Patton and An. gambiae Giles sensu stricto (Diptera: Culicidae) were investigated for interspecific competition. Single-species and mixed-species populations were reared at 27 degrees C from the first instar to pupation at different densities (100, 200 or 400 larvae/200 cm2 tray) with a constant amount of food, 0.2 mg/larva/day. Pupae obtained from mixed populations were identified to species using PCR. Both species had a 1:1 sex ratio at pupation. Development time to pupation averaged about one day less for An. arabiensis compared to An. gambiae, ranging from 0.93-1.49d for males and from 0.44-0.84 d for females in single populations. In mixed species populations the difference for males ranged from 0.99-1.58d and for females from 0.93-1.62d. Survival rates of An. gambiae s.s. were significantly higher than those of An. arabiensis in both the single-species and mixed-species populations. Mixed-species rearing did not have an effect on the survival of An. gambiae, whereas the mortality rate of An. arabiensis was significantly higher in mixed populations than when only this species was reared at the same densities, suggesting a competitive disadvantage for An. arabiensis in mixed populations. High proportions of larvae (4-35%) were lost during development; these losses could not be accounted for by corpses found in the rearing pans. The possibility of cannibalism was investigated by rearing each species separately in small containers (five per 50 ml), inspected every 6h, but no cannibalism was detected at any stage of development in either species. It was concluded that, under these experimental circumstances, interspecific competition between both species did occur but with a detrimental effect on An. arabiensis only. Relevance of these findings to the ecology of both species in the field is discussed briefly.     

Assortative mating in mixed swarms of the mosquito Anopheles gambiae s.s. M and S molecular forms,in Burkina Faso,West Africa

The molecular form composition of Anopheles gambiae Giles s.s. (Diptera: Culicidae) mating swarms and the associated mating pairs (copulae) were investigated during two rainy seasons (July to October, 2005 and July to November, 2006) in the villages of Soumousso and Vallée du Kou (VK7). Although the habitats of these villages differ markedly, sympatric populations of M and S molecular forms of An. gambiae s.s. occur in both places periodically. The main aim was to assess the degree to which these molecular forms mate assortatively. In Soumousso, a wooded savannah habitat, the majority of swarm samples consisted of only S‐form males (21/28), although a few M‐form males were found in mixed M‐ and S‐form swarms. In VK7, a rice growing area, the majority of swarm samples consisted of only M‐form males (38/62), until October and November 2006, when there were nearly as many mixed‐form as single‐form swarms. Overall, ～60% of M‐ and S‐form swarms were temporally or spatially segregated; the two forms were effectively prevented from encountering each other. Of the remaining 40% of swarms, however, only about half were single‐form and the rest were mixed‐form. Of the 33 copulae collected from mixed‐form swarms, only four were mixed‐form pairs, significantly fewer than expected by random pairing between forms (χ² = 10.34, d.f. = 2, P < 0.01). Finally, all specimens of inseminated females were of the same form as the sperm contained within their spermatheca (n = 91), even for the four mixed‐form copulae. These findings indicate that assortative mating occurs within mixed‐form swarms, mediated most probably by close‐range mate recognition cues.     

Anopheles arabiensis and An. quadriannulatus resistance to DDT in South Africa

The malaria control programme of KwaZulu‐Natal Province, South Africa, includes Mamfene and Mlambo communities. Western‐type houses there are currently sprayed with deltamethrin, whereas traditional houses are sprayed with DDT for malaria control. In 2002, mosquitoes of the Anopheles gambiae complex (Diptera: Culicidae) were collected from DDT‐sprayed houses, by window exit traps, and from man‐baited nets outdoors. Larval collections were also carried out at Mzinweni Pan near Mlambo. Species of the An. gambiae complex were identified by rDNA polymerase chain reaction assay. The majority of samples collected by window trap and baited nets were identified as the malaria vector An. arabiensis Patton, with a few An. merus Dönitz and An. quadriannulatus (Theobald). The larval collections were predominantly An. quadriannulatus with a small number of An. arabiensis. Standard WHO insecticide susceptibility tests using 4% DDT and 0.05% deltamethrin were performed on both wild‐caught females and laboratory‐reared progeny from wild‐caught females. Wild‐caught An. arabiensis samples from window traps gave 63% and 100% mortality 24‐h post‐exposure to DDT or deltamethrin, respectively. Wild‐caught An. arabiensis samples from man‐baited net traps gave 81% mortality 24‐h post‐exposure to DDT. The F₁ progeny from 22 An. arabiensis females showed average mortality of 86.5% 24‐h post‐exposure to DDT. Less than 80% mortality was recorded from five of these families. Biochemical analyses of samples from each of the families revealed comparatively high levels of glutathione‐S‐transferases and non‐specific esterases in some families, but without significant correlation to bioassay results. Wild‐caught An. quadriannulatus larvae were reared through to adults and assayed on 4% DDT, giving 47% (n = 36) mortality 24‐h post‐exposure. Finding DDT resistance in the vector An. arabiensis, close to the area where we previously reported pyrethroid‐resistance in the vector An. funestus Giles, indicates an urgent need to develop a strategy of insecticide resistance management for the malaria control programmes of southern Africa.     

The demography of Howler Monkeys (Alouatta palliata) on Barro Colorado Island,Panamá

Chronological ages of Alouattaon Barro Colorado Island (BCI) were estimated from longitudinal dental wear. Combining these data with visual censuses, we approximated the study population’s age profile. A stable model was then constructed; from it we derived agespecific mortality rates. Mortality of immature animals is high, with 88% of the males and 65% of the females dying before 5 years. Adult mortality is low until 11 years, when it accelerates. The average adult life span is 16.6 years for males and 15.5 for females. The maximum life span is over 20 years. A pubertal male growth spurt occurs from 3 to 5 years, at which time females are primiparous. Sexual size differences develop primarily during this time. The 1976 age profile had anomalously few animals aged 7 years and males aged 8–9 and 15–16 years. With corroborating evidence, we hypothesize that these deficits resulted from excessive rainfall in 1963 and 1971, which reduced the fall fruit crop and led to a high juvenile mortality. Within half of our study troops, some adult males have nearly identical ages, suggesting an active process of agemate coalition. We hypothesize a form of kin selection, wherein peripheral male cohorts from the same natal troop have greater survival and social success than solitary animals. The stable model suggests an annual growth rate of 1.5% during the 1970s. Estimates of 16.7 and 4% for the previous two decades, following a yellow-fever epidemic, imply that the BCI population is becoming stationary. A very different age structure existed on neighboring Orchid Island in 1976, suggesting general food limitation. By contrast, the lowered, but continuing growth on BCI suggests a less drastic limiting mechanism. We hypothesize that intratroop social competition limits population growth by regulating subadult survival rates during food scarcity cycles.