Sporozoite Rate Research Articles

Baseline Entomological Data for a Pilot Malaria Control Program in Equatorial Guinea

Entomological studies conducted on the island of Bioko and the mainland province, Río Muni, in Equatorial Guinea provided baseline data for a pilot malaria control program. The main species collected resting in houses at low density were Anopheles gambiae s.l. Giles and Anopheles funestus Giles. An. gambiae s.l. density increased during the rainy season, whereas An. funestus populations remained stable during both dry and wet seasons. Both species tended to be endophilic and were not collected resting or biting outdoors. Overall, delayed sporozoite rates were about 10% for both species and were highest in the dry season. An. gambiae s.l. from the mainland showed resistance to 4% DDT, but the island population was still susceptible. All populations were completely susceptible to 5% malathion and 0.025% deltamethrin.

Malaria in urban and rural Kinshasa: the entomological input.

Mosquitoes were collected on human bait over a 16-month period (September 1988 to December 1989) in an urban and a rural area of Kinshasa, Zaïre. P. falciparum malaria sporozoite rates were determined by ELISA. In the urban area Culex quinquefasciatus accounts for 96% of the 121 bites/person/night (b/p/n). The only anopheline is Anopheles gambiae, sensu stricto, with an average of 5.1 b/p/n and a sporozoite rate of 1.86%. The entomological inoculation rate (EIR) averages 0.08 infective b/p/n. Malaria transmission is almost interrupted at the end of the dry season. In the rural area mosquito nuisance is small (20 b/p/n), almost entirely due to six species of Anopheles including four vectors of malaria: An.gambiae (13.3 b/p/n), An.funestus (2.4 b/p/n), An.nili (0.4 b/p/n) and An.brunnipes (0.7 b/p/n) with mean sporozoite rates of 7.85%, 6.60%, 6.63% and 0.53% respectively. An.paludis (0.4 b/p/n) and An.hancocki (0.2 b/p/n) were not found infective. Malaria transmission is intense and perennial: the overall EIR varies monthly between 0.60 and 3.29 infective b/p/n. The specific contributions of An.gambiae, An.funestus and An.nili average 1.07, 0.14 and 0.03 infective b/p/n respectively. Malaria transmission peaks during the rainy season in both study areas. The daily mean survival rates for An.gambiae were 0.91 and 0.78 in the rural and urban area, respectively. All An.gambiae examined belonged to the forest cytotype (Coluzzi et al., 1979). Through its effect on the sporozoite rate, the higher vector survival rate in the rural environment appears to be the major determinant of the greater malaria transmission rate in the rural area as compared to urban Kinshasa.

Plasmodium falciparum sporozoite and entomological inoculation rates at the Ahero rice irrigation scheme and the Miwani sugar-belt in western Kenya.

Anopheles arabiensis and An. funestus were collected by pyrethrum spray sheet collections in houses and by human-bait catches at a village in western Kenya adjacent to the Ahero rice irrigation scheme; and using the same methods, An. gambiae s.l. and An. funestus were collected at Miwani, a village in the sugar-cane belt. Plasmodium falciparum sporozoite rates were determined by ELISA. At Ahero the mean sporozoite rates were 1.1% and 4.3% in An. arabiensis and An. funestus, respectively, while at Miwani the rates were 6.0% in An. gambiae s.l. and 4.3% in An. funestus. Entomolgoical inoculation rates (EIR) were derived from both human-bait collections (IR-HBC) and by the proportion of human blood-fed females caught resting indoors (IR-HBF). The IR-HBF appeared to be a more realistic index of EIR. At Ahero and Miwani people were exposed to an average of 416 and 91 infective bites/person/year, respectively. The main vectors were An. funestus at Ahero and An. gambiae s.l. at Miwani. In view of the intense and perennial malaria transmission at Ahero, vector control by insecticides should be considered, while at Miwani, where transmission is seasonal, permethrin-impregnated bed nets could be an alternative to indoor spraying. These measures must be augmented with availability of effective antimalarials.

Entomological risk factors for severe malaria in a peri-urban area of The Gambia.

A study was undertaken of possible entomological risk factors for severe malaria in a peri-urban area of The Gambia. Households of children who had experienced a severe or a mild attack of malaria and of matched controls were visited and their characteristics recorded. Mosquitoes were then collected in the bedrooms of study subjects using both insecticide spray catches and light traps. Mud-walled buildings and bedrooms without ceilings were found more frequently in the households of children who had experienced malaria than in those of controls. Only one difference, which may have arisen by chance, was found between the households of severe and of mild malaria cases; animals were recorded significantly more frequently in the compounds of the severe cases. Mosquito catches reported on two occasions 2 weeks apart showed good reproducibility, indicating that measurements made at the time that a child presented with malaria were likely to reflect those that would have been present at the time that the child was infected. Anopheles gambiae s.l. was found on average more frequently in the bedrooms of children who had experienced malaria than in those of the controls but no difference was found in mosquito numbers between bedrooms of severe or mild cases of malaria. The human blood index and sporozoite rate were similar in mosquitoes caught in households of malaria cases and in those of controls. No evidence was found to support the hypothesis that the level of exposure to malaria-infected mosquitoes is a risk factor for the development of severe malaria.

Ookinete Rates in Afrotropical Anopheline Mosquitoes as a Measure of Human Malaria Infectiousness

Anopheles gambiae s.1. and An. funestus were sampled for Plasmodium spp. ookinetes in two P. falciparum-endemic sites in western Kenya. Since the ookinete is a transitional stage of short duration, occurring after fertilization and before oocyst development, only females in the half-gravid and gravid stages of blood digestion were examined. Preparations of homogenized midguts were spotted onto microslides and examined microscopically after staining with Giemsa. Overall, ookinetes were detected in 4.4% of 1,079 anophelines examined over an eight-month period. Anopheles funestus had higher ookinete rates than An. gambiae s.1., and ookinete rates were higher in half-gravid than in gravid An. gambiae s.1. Geometric mean numbers of ookinetes per infected female were less than five for each species at the two sites, and the maximum number observed was only 12. The low frequencies and numbers of ookinetes were sufficient to produce sporozoite rates of 4-18% in the vector populations. The intense transmission of P. falciparum in these two sites is maintained by anthropophilic vectors where only one in 23 blood meals initiates an infection of generally less than five ookinetes. Relationships between human malaria infectiousness and vector infectivity are dependent upon the high efficiency of the developmental transition from the ookinete to the subsequent oocyst and sporozoite stages.

Malaria transmission and vector biology on Sainte Marie Island, Madagascar.

A 17-mo longitudinal malaria survey (November 1988-March 1990) was carried out on Sainte Marie Island, an area on the east coast of Madagascar which is frequently visited by tourists. During 706 man-nights of capture, 46,401 mosquitoes belonging to 32 species were caught. Sporozoite rates were determined by ELISA and incriminated Anopheles gambiae Giles s.s., An. funestus Giles, and An. mascarensis De Meillon as vectors of malaria. An. gambiae, the main vector, was highly anthropophilic but largely exophilic. Transmission by this species occurred mainly from November to April; the overall circumsporozoite antigen positivity rate was 1.7%, with a maximum of 3.2% in March-April. The nightly peak of transmission occurred between midnight and 0400 hours. The annual inoculation rate was calculated to be 100 infective bites per human, of which 92 were of Plasmodium falciparum. An. funestus played a minor role in transmission. An. mascarensis, an anopheline endemic to Madagascar, was incriminated for the first time, as a malaria vector. The sporozoite rate in this species varied from 0.4 to 0.9% as shown by both ELISA and salivary gland dissections. Parasite indices in humans up to 20 yr of age fluctuated during the year from 64 to 80%. Bed nets are recommended for malaria protection for the local population and tourists.

Method to estimate relative transmission efficiencies of Anopheles species (Diptera: Culicidae) in human malaria transmission.

A mathematical expression was derived to estimate the relative malaria transmission efficiency of an anopheline species with respect to a standard well-characterized species for which all vector parameters can be sufficiently determined. The method is particularly useful in situations where multiple anopheline species contribute to human malaria transmission and requires the estimation of the man-biting rate, the sporozoite rate, and the human malaria incidence. Under stable conditions of vector abundance, the average sporozoite rate in a species during a transmission season would by itself reflect its relative transmission efficiency. This "efficiency" then was used to calculate the "effective human-biting rate"; i.e., the human-biting rate of that species if it were to have ecological properties identical to those of the standard species. The standard well-characterized species then could be used with the effective human-biting rate of all species to quantify transmission, thus overcoming the need to measure vector parameters for all anopheline species contributing to transmission. An expression also was derived to calculate the relative contribution made by each species to malaria transmission. The usefulness of this method was illustrated using entomological and epidemiological data from Kataragama, Sri Lanka.

Trial of pyrethroid impregnated bednets in an area of Tanzania holoendemic for malaria Part 2. Effects on the malaria vector population

The malaria vector population consisted mainly of Anopheles gambiae s.s. with a small contribution from An. funestus and An. rivulorum. The mosquitoes coming to bite in bedrooms were monitored with light traps set beside untreated bednets. When impregnated bednets were provided for all the other beds in a village the Anopheles populations declined but the Culex quinquefasciatus populations were unaffected. The survival of An. gambiae (as measured by the mean number of ovarian dilatations) and the sporozoite rate declined following introduction of the nets and the estimated sporozoite inoculation rates into people not under their nets declined by more than 90%. The net introductions caused sharp declines in the number of mosquitoes resting indoors, but the evidence was inconclusive regarding diversion to outdoor resting, animal biting, earlier biting or outdoor biting. DDT spraying greatly reduced the Anopheles populations.

Human malaria infectiousness measured by age-specific sporozoite rates in Anopheles gambiae in Tanzania

In an area of holoendemic malaria in Northern Tanzania, Anopheles gambiae s.l. females were age-graded by Polovodova's method and dissected for sporozoites. Age-specific sporozoite rates implied that mosquitoes acquired new infections at all ages. The extrinsic period lasted just over 3 gonotrophic cycles (9-11 days). Very high sporozoite rates in the oldest females implied the absence or rarity of genetic refractoriness to infection. A method is described for estimating the proportion of bloodmeals which result in mosquito infection. This method makes relatively few assumptions about mosquito behaviour, and could be useful for evaluating transmission-blocking interventions. Overall, it is estimated that about 21% of meals are infectious. This is much higher than previous estimates derived either from experimental mosquito feeding studies or from similar age-grading data collected from the same area in 1962. Various alternative explanations are considered, and it is concluded that there has been a 2.5-fold increase in human infectiousness in the last 25 years. This is partly attributable to suppression of human infectiousness by widespread chloroquine usage during the 1960s, followed by removal of this effect by drug resistance. It is argued that chloroquine would be expected to select for increased infectivity in the parasite, and this may also have contributed to the observed increase.

Monitoring human-biting mosquitoes (Diptera: Culicidae) in Tanzania with light-traps hung beside mosquito nets

AbstractMosquitoes were caught in bedrooms in Tanzanian villages by human-biting catches and in light-traps set close to occupied untreated bed nets. Catches by each method were carried out on pairs of nights in the same week at different seasons and in different villages. The pairs of adjacent catches by the different methods showed a strong correlation. Analysis of the ratio between the catches by the two methods on pairs of nights in the same week indicated that on average three light-traps caught about the same number of mosquitoes as a team of two human catchers. The ratio did not differ significantly between Anopheles gambiae Giles (sensu lato), A. funestus Giles, and Culex quinquefasciatus Say, nor between the villages, or between times when mosquito populations were high or low. The distribution of numbers of ovarian dilatations differed significantly between catches in different villages and seasons but not between pairs of catches by the two methods. Similarly, the parity and sporozoite rates agreed between pairs of light-trap and house-resting catches, but differed markedly between villages and seasons. Thus it is concluded that light-traps used in conjunction with bed nets catch a representative sample of the vectors which would have bitten humans in bedrooms in this area.

Comparative determination ofPlasmodium falciparumsporozoite rates in AfrotropicalAnophelesfrom Kenya by dissection and ELISA

The Plasmodium falciparum rate was determined by microscopical examination of one salivary gland (three lobes) and by enzyme-linked immunosorbent assay (ELISA) of the other salivary gland in each of 1580 Anopheles mosquitoes collected from western Kenya during both the wet and dry seasons. The sporozoite rate in the wet season was much higher than that in the dry season, and the sporozoite rate determined by ELISA was generally lower than that determined by microscopy. The ELISA gave a positive reaction to circumsporozoite protein in some glands whose counterparts did not show the presence of sporozoites by microscopy, thus giving an 'overestimation' of the sporozoite rate. This overestimation was greater in the dry season than in the wet season, and greater in Anopheles gambiae than in An. funestus, but overall it was only 1.2% (19/1580). These results are at variance with reports of other workers, who have shown ELISA overestimation of the sporozoite rate as high as 30%. Our tests indicated that the ELISA sensitivity was 80.6%, its specificity was 98.7%, and its accuracy was 97.5%.

Anatomical dissemination of circumsporozoite protein in wild Afrotropical Anopheles affects malaria sporozoite rate determination by ELISA

The head, thorax, wings, legs and abdomen of 320 wild-caught Anopheles gambiae Giles sensu lato and 115 An.funestus Giles were tested by an enzyme-linked immunosorbent assay (ELISA) for Plasmodium falciparum Welch to determine how anatomical dissemination of circumsporozoite (CS) protein could affect the estimation of malaria sporozoite rates by ELISA. Of fifty-three Anopheles with CS protein detected in any body part, positive reactions were observed for 58.5% of heads, 67.0% of thoraces, 39.6% of wings, 52.8% of legs and 60.4% of abdomens. Mean absorbance values (range 0-2.00) were highest in thorax samples (1.17), followed by heads (0.80), abdomens (0.67), wings (0.48) and legs (0.46). Circumsporozoite protein was present in the wings or legs, but not in the head or thorax, in 11.3% (6/53) of the infected Anopheles. The ELISA infection rate of 12.8% (41/320) for An.gambiae would have increased to 14.7% (47/320) by inclusion of six mosquitoes with CS protein in wings or legs alone. The slight overestimation of the proportion of infective mosquitoes due to disseminated CS protein would have little effect on estimates of relative infection rates by ELISA for field-collected Anopheles, with abdomens removed prior to testing. However, the widespread dissemination of CS protein indicates that sporozoite load estimates by ELISA, for mosquitoes without abdomens, may not provide adequate measurements of the numbers of sporozoites in the salivary glands. Operationally, careful processing of mosquito samples for the determination of infectivity rates by ELISA is necessary to prevent the mixing of wings or legs among samples representing individual mosquitoes.

Malaria transmission potential of Anopheles mosquitoes in the Mwea-Tebere irrigation scheme, Kenya

1. Anopheles arabiensis Patton and An. funestus Giles were identified as vectors of Plasmodium falciparum malaria in the Mwea-Tebere irrigation scheme, Kenya. An. arabiensis was the only member of the An. gambiae complex identified from chromosome characteristics. Other Anopheles species found included An. pharoensis Theobald, An. rufipes Gough and An. coustani Laveran. Survival rates per gonotrophic cycle for An. arabiensis averaged 0.37 during the short rains (October-November), 0.49 during the dry season (February) and 0.78 during the long rains (May-June). Vectorial capacities were correspondingly low due to low survival rates and a high degree of zoophily. The average duration of infective life for P. falciparum was 0.2 days for both An. arabiensis and An. funestus. In contrast, entomological inoculation rates were comparatively high: 6-8 infective bites/man/month. An. pharoensis averaged 110 bites/man/night during the short rains; 1/999 (0.1%) was positive by ELISA for P. falciparum circumsporozoite antigen, but the ELISA evidence is not conclusive for vector incrimination. In correspondence with clinical observations, the transmission of P. malariae and P. ovale is unlikely due to the low vector survival rates. The observed anomaly between low vectorial capacities and high entomological inoculation rates demonstrates the importance of accurately estimating vector sporozoite rates to monitor unstable malaria transmission in irrigated areas.

Malaria Transmission and Vector Biology in Manarintsoa, High Plateaux of Madagascar

To evaluate the factors which determine the transmission level of falciparum malaria, entomological and parasitological surveys were conducted from October 1988 to February 1990 in Manarintsoa in the central highland plateaux of Madagascar. Mosquitoes were collected for 928 man-nights in pit shelters and indoor resting sites. Malaria vectors were Anopheles arabiensis and An. funestus, with no evidence of the presence of An. gambiae sensu stricto. Vectors were mainly exophilic and zoophilic. The index of stability was less than 1.5. The sporozoite rate was 0.11 for An. gambiae sensu lato and 0.47 for An. funestus. The transmission level was low, with an inoculation rate of 0.91 infective bites/person/year and an infection risk of 0.62. Malaria transmission occurs 7 months of the year in this area, from November to May. Human parasite rates fluctuated from 29% in October to 53% in May.

Characterization of Malaria Transmission by Anopheles (Diptera: Culicidae) in Western Kenya in Preparation for Malaria Vaccine Trials

Malaria transmission was studied for 33 mo in the villages of Kisian and Saradidi in western Kenya in preparation for field trials of malaria vaccines. Abundance estimates of Anopheles gambiae Giles sensu lato and Anopheles funestus Giles, which constituted over 99% of 26,645 anophelines collected, were compared for all-night biting collections inside houses, outdoors, and in tents. The overall numbers of Anopheles per man-night were 2.3 times greater in Kisian than in Saradidi. For the three types of collections, mean sporozoite rates by dissection ranged from 2.2 to 5.4% for 13,072 Anopheles in Kisian and from 9.9 to 13.6% for 7,058 Anopheles in Saradidi; greater than 90% of the infections were Plasmodium falciparum, either alone or mixed with P. malariae or P. ovale. Heaviest transmission from April to July coincided with the end of the long rainy season. Entomological inoculation rates (EIR) averaged 0.82 infective bites per man per night inside houses in Kisian and 0.65 in Saradidi. Outdoors, EIRs averaged 0.09 in Kisian and 0.52 in Saradidi. In tents, which were evaluated to identify methods for exposing nonindigenous volunteers during vaccine efficacy trials, EIRs were 3.3 and 2.5 times less than inside houses for Kisian (EIR = 0.25) and Saradidi (EIR = 0.26), respectively. Exposure in tents averaged one infective bite every 4.0 d in Kisian and every 3.8 d in Saradidi. The use of tents in vaccine efficacy trials should provide adequate exposure for nonindigenous volunteers. Malaria vaccine trials could be conducted efficiently in western Kenya, with timing dependent upon the intensity of transmission required by vaccine trial objectives.

Malaria Sporozoite Detection by Dissection and Elisa to Assess Infectivity of Afrotropical Anopheles (Diptera: Culicidae)

Malaria infection rates determined by dissection and Plasmodium falciparum enzyme-linked immunosorbent assay (ELISA) were compared for 26,935 Anopheles gambiae Giles sensu lato and 17,739 Anopheles funestus Giles collected during 20 mo in western Kenya. ELISA infection rates were about 43% higher than dissection sporozoite rates. In dissection-negative Anopheles, circumsporozoite (CS) protein was detected by ELISA in 5.2% of 10,017 salivary gland samples and in 12.2% of 237 thorax samples. The accuracy of dissection and ELISA techniques was compared by the following tests on a group of 352 field-collected Anopheles (held 10 d to ensure sporogonic development): salivary gland dissection, examination of Giemsa-stained dissection slides, ELISA tests on salivary gland and thorax body parts, and microscopic techniques for determining sporozoite loads. Respective infection rates were 9.9%, 10.8%, and 15.6% for dissection, stained slides, and ELISA. Sporozoite loads were associated significantly with ELISA absorbance values (r = 0.76). Compared with Giemsa-stained dissection slide results, the sensitivity of sporozoite detection was 92.1% for dissection compared with 78.9% for ELISA; specificity was 100.0% for dissection versus 92.0% for ELISA. Immunological detection of CS protein in head-thorax samples of Afrotropical vectors overestimated the proportion of infective Anopheles because the comparison of techniques indicated that 45.4% of the ELISA positive Anopheles did not contain salivary gland sporozoites.

Effect of human circumsporozoite antibodies in Plasmodium-infected Anopheles (Diptera: Culicidae).

Human circumsporozoite (CS) antibodies to Plasmodium falciparum were detected in blood meals from 45.0% of 1,547 field-collected Anopheles gambiae Giles sensu lato and Anopheles funestus Giles from western Kenya. Possible effects on malaria infections within the Anopheles host were investigated. Circumsporozoite antibodies were detected in blood meals up to 36 h after feeding. Antibodies crossing the midgut were detected experimentally in hemolymph from 4 to 36 h after feeding; human IgG also was present in hemolymph from fully gravid field-collected Anopheles. Ingestion of high-titer human CS antibodies or 2A10 monoclonal antibody to P. falciparum sporozoites by P. falciparum-infected An. gambiae, 10 d after feeding on an infected human, had no effect on oöcyst maturation, sporozoite rates, or sporozoite loads. Contact between CS antibodies and sporozoites in the hemocoel did not block sporozoite invasion of salivary glands. Human IgG antibodies were detected by an indirect fluorescent antibody technique on salivary gland sporozoites from 83.3% of 114 field-collected Anopheles. In 65.4% of 26 infections, antibodies persisted on sporozoites for at least three days. Thus, a high proportion of naturally infected An. gambiae s.l. and An. funestus in western Kenya transmit sporozoites that are bound with human IgG acquired during previous blood meals. The infectivity of such sporozoites needs to be determined in relation to natural transmission and to the potential use of malaria sporozoite vaccines.

Application of a biochemical key to study transmission of malaria and bancroftian filariasis in sibling species of the Anopheles gambiae complex in north-eastern Tanzania

A biochemical key was applied in order to study transmission of malaria and Bancroftian filariasis in Anopheles gambiae sensu stricto, An. arabiensis and An. merus in different localities in north-eastern Tanzania. The technique was found to be a useful additional taxonomic tool for field entomologists. Significant differences between species in the rate of infection with Bancroftian filariasis were obtained between An. gambiae s.s. and An. funestus ( P<0.005) and between An. funestus and An. arabiensis ( P<0.0001). There were also significant differences between most of the investigated localities in the rate of filarial infection. However, there were no significant differences between the three species or between localities with respect to malaria sporozoite rates. Possible reasons for the observed variation between species and localities with respect to vectorial activity for Bancroftian filariasis are discussed.

Seasonal population changes and malaria transmission potential of Anopheles pharoensis and the minor anophelines in Mwea Irrigation Scheme, Kenya

A study in 1984 and 1985 showed that Anopheles gambiae s.l. and An. pharoensis were the major anophelines in Mwea Irrigation Scheme, Kenya, constituting 83.86% and 15.69% of the catch respectively. Four minor species made up the remaining 0.45%. The irrigation phase of the rice cultivation cycle in August, which linked the flooding effects of the two rainy seasons, resulted in major population increases of An. pharoensis and enabled continuous breeding for up to 9 months per year. The average of mean monthly proportions of unfed, bloodfed, and gravid females was 26.6, 58.8, and 14.6% respectively. The Plasmodium falciparum sporozoite rates for An. pharoensis were 1.3% by ELISA and 0.68% by dissection, while those for An. funestus were 1.7% by ELISA and 1.25% by dissection. An. pharoensis can contribute to the epidemiology of Malaria in the Mwea area.

An analysis of some factors determining the sporozoite rates, human blood indexes, and biting rates of members of the Anopheles punctulatus complex in Papua New Guinea.

The degree to which Anopheles punctulatus complex members feed on humans in different Papua New Guinea villages has a significant effect on sporozoite rates. Among villages, the human blood index (HBI) of the members of the complex varied with the average number of persons sharing a bednet. Although dogs are the preferred hosts by the 3 malaria vector species, the number of dogs did not significantly affect the HBI. The HBI was dependent upon the human-biting rate, implying increased avoidance of anophelines by people relative to other hosts at times of greater mosquito numbers. Human-biting rates and HBIs were also influenced by the distribution of alternative hosts relative to people.