Sporozoite Rate Research Articles

Malaria entomological inoculation rates in gold mining areas of Southern Venezuela

A longitudinal study of malaria vectors aiming to describe the intensity of transmission was carried out in five villages of Southern Venezuela between January 1999-April 2000. The man-biting, sporozoite and entomological inoculation rates (EIR) were calculated based on 121 all-night collections of anophelines landing on humans, CDC light traps and ultra violet up-draft traps. A total of 6,027 female mosquitoes representing seven species were collected. The most abundant species were Anopheles marajoara Galvão & Damasceno (56.7%) and Anopheles darlingi Root (33%), which together accounted for 89.7% of the total anophelines collected. The mean biting rate for An. marajoara was 1.27 (SD + 0.81); it was 0.74 (SD + 0.91) for An. darlingi and 0.11 (SD + 0.10) for Anopheles neomaculipalpus Curry and the overall biting rate was 2.29 (SD + 1.06). A total of 5,886 mosquitoes collected by all three methods were assayed by ELISA and 28 pools, equivalent to 28 mosquitoes, yielded positive results for Plasmodium spp. CS protein. An. neomaculipalpus had the highest sporozoite rate 0.84% (3/356), followed by An. darlingi 0.82% (16/1,948) and An. marajoara 0.27% (9/3,332). The overall sporozoite rate was 0.48% (28/5,886). The rates of infection by Plasmodium species in mosquitoes were 0.37% (22/5,886) for Plasmodium vivax(Grassi & Feletti) and 0.10% (6/5,886) for Plasmodium falciparum (Welch). The estimated overall EIR for An. darlingi was 2.21 infective bites/person/year, 1.25 for An. marajoara and 0.34 for An. neomaculipalpus. The overall EIR was four infective bites/person/year. The biting rate, the sporozoite rate and the EIR are too low to be indicators of the efficacy of control campaigns in this area.

Implementation of a novel PCR based method for detecting malaria parasites from naturally infected mosquitoes in Papua New Guinea

BackgroundDetection of Plasmodium species in mosquitoes is important for designing vector control studies. However, most of the PCR-based detection methods show some potential limitations. The objective of this study was to introduce an effective PCR-based method for detecting Plasmodium vivax and Plasmodium falciparum from the field-caught mosquitoes of Papua New Guinea.MethodsA method has been developed to concurrently detect mitochondrial cytochrome b (Cyt b) of four human Plasmodium species using PCR (Cytb-PCR). To particularly discriminate P. falciparum from P. vivax, Plasmodium ovale and Plasmodium malariae, a polymerase chain reaction-repeated fragment length polymorphism (PCR-RFLP) has further been developed to use with this method. However, due to limited samples number of P. ovale and P. malariae; this study was mainly confined to P. vivax and P. falciparum. The efficiency of Cytb-PCR was evaluated by comparing it with two 'gold standards' enzyme linked immunosorbent assay specific for circumsporozoite protein (CS-ELISA) using artificially infected mosquitoes; and nested PCR specific for small subunit ribosomal RNA (SSUrRNA) using field caught mosquitoes collected from three areas (Kaboibus, Wingei, and Jawia) of the East Sepic Province of Papua New Guinea.ResultsA total of 90 mosquitoes were artificially infected with three strains of Plasmodium: P. vivax-210 (n = 30), P. vivax-247 (n = 30) and P. falciparum (n = 30). These infected mosquitoes along with another 32 unfed mosquitoes were first checked for the presence of Plasmodium infection by CS-ELISA, and later the same samples were compared with the Cytb-PCR. CS-ELISA for P. vivax-210, P. vivax-247 and P. falciparum detected positive infection in 30, 19 and 18 mosquitoes respectively; whereas Cytb-PCR detected 27, 16 and 16 infections, respectively. The comparison revealed a close agreement between the two assays (κ = 0.862, 0.842 and 0.894, respectively for Pv-210, Pv-247 and P. falciparum groups). It was found that the eight CS-ELISA-positive mosquitoes detected negative by Cytb-PCR were false-positive results. The lowest detection limit of this Cytb-PCR was 10 sporozoites. A highly concordance result was also found between nested PCR and Cytb-PCR using 107 field caught mosquitoes, and both tests concordantly detected P. falciparum in an Anopheles punctulatus mosquito collected from Kaboibus. Both tests thus suggested an overall sporozoite rate of 0.9% (1/107) in the study areas. Subsequently, PCR-RFLP efficiently discriminated P. falciparum from P. vivax for all of the Cytb-PCR positive samples.ConclusionA single step PCR based method has been introduced here that is highly sensitive, efficient and reliable for identifying P. vivax and P. falciparum from mosquitoes. The reliability of the technique was confirmed by its ability to detect Plasmodium as efficiently as those of CS-ELISA and nested PCR. Application of the assay offers the opportunity to detect vector species of Papua New Guinea and may contribute for designing further vector control programmes.

Good News in Malaria Control… Now What?

1describe remarkable malaria control progress and benefits on the island of Bioko in Equatorial Guinea over the last 4 years. Using a pre-program time period as their comparison, they show that between 2004–2008 with high household coverage of twiceyearly indoor residual spraying (IRS), the introduction of artemisinin combination treatment (ACTs), the use of intermittent preventive treatment in pregnant women (IPTp), and the somewhat more recent introduction of very high coverage with long-lasting insecticide treated mosquito nets (LLINs), they had remarkable impact on malaria infection, disease, and mortality in the population. Although they present their findings fully in their article, some merit repeating. First, their efforts achieved 4 years of very good coverage: spraying IRS every 6 months in typically > 80% of households; achieving 73% LLIN use (not just ownership) in households through house-to-house distribution with instruction and education all provided at no cost to the people; widespread training of doctors and nurses, routine use of diagnostics and free first-line treatment of children 90% and reduced sporozoite rates in the remaining mosquitoes by > 90%. Infection prevalence in 2- to 5-year-old children was down ~57% to 18%; fever rates in the previous 4 weeks were down ~56% to 6.3%; ACT treatment needs decreased by ~62%; anemia rates (Hb < 8 gm/dL) in 2 to 5 year olds were down ~87% to just 2 %. This progress appears to have contributed substantially to a reduction in all-cause under-five mortality from 152 → 55 deaths per 1,000 live births (down ~64%); and the drop occurred rapidly and timed directly with the intervention scale-up. These are findings from a program effort, albeit well monitored and supported with national contributions and much external support from experts and donors. And, the authors must rely on before-scale-up and after-scale-up comparisons. As a consequence, the authors document as well as possible the other potential contributions to improved child survival. The improved economy in Equatorial Guinea caused by the oil industry is explored and the slightly lower rainfall during the 2005–2008 program years is examined in their analysis. They looked for other substantial improvements in disease control efforts (e.g., immunization coverage, other maternal or child health interventions) and found no dramatic scaleup of these programs during this interval to account for their findings. In summary, scaled up malaria control with currently available interventions works; it reduces vector populations and their transmission potential, lowers infection rates, illness rates, anemia rates, and saves lives. This is consistent with the controlled trials for some of the interventions 2–5

The susceptibility of Anopheles lesteri to infection with Korean strain of Plasmodium vivax

BackgroundFollowing its recent re-emergence, malaria has gained renewed attention as a serious infectious disease in Korea. Three species of the Hyrcanusgroup, Anopheles lesteri, Anopheles sinensis and Anopheles pullus, have long been suspected malaria vectors. However, opinions about their vector ability are controversial. The present study was designed with the aim of determining the susceptibility of these mosquitoes to a Korean isolate of Plasmodium vivax. Also, An. sinensis is primarily suspected to be vector of malaria in Korea, but in Thailand, the same species is described to have less medical importance. Therefore, comparative susceptibility of Thai and Korean strains of An. sinensis with Thai strain of P. vivax may be helpful to understand whether these geographically different strains exhibit differences in their susceptibility or not.MethodsThe comparative susceptibility of An. lesteri, An. sinensis and An. pullus was studied by feeding laboratory-reared mosquitoes on blood from patients carrying gametocytes from Korea and Thailand.ResultsIn experimental feeding with Korean strain of P. vivax, oocysts developed in An. lesteri, An. sinensis and An. pullus. Salivary gland sporozoites were detected only in An. lesteri and An. sinensis but not in An. pullus. Large differences were found in the number of sporozoites in the salivary glands, with An. lesteri carrying much higher densities, up to 2,105 sporozoites in a single microscope field of 750 × 560 μM, whereas a maximum of 14 sporozoites were found in any individual salivary gland of An. sinensis. Similar results were obtained from a susceptibility test of two different strains of An. sinensis to Thai isolate of P. vivax, and differences in vector susceptibility according to geographical variation were not detected.ConclusionThe high sporozoite rate and sporozoite loads of An. lesteri indicate that this species is highly susceptible to infection with P. vivax. Anopheles sinensis appears to have a markedly reduced ability to develop salivary gland infection, whilst in An. pullus, no sporozoites were found in the salivary glands. Provided that the survival rate of An. lesteri is sufficiently high in the field, it would be a highly competent vector of vivax malaria.

Transmission dynamics of malaria in four selected ecological zones of Nigeria in the rainy season

Two of the problems of malaria parasite vector control in Nigeria are the diversity of Anopheline vectors and large size of the country. Anopheline distribution and transmission dynamics of malaria were therefore compared between four ecotypes in Nigeria during the rainy season. Polymerase chain reaction (PCR) was used in molecular identification after morphological identification microscopically. Enzyme linked immunosorbent assay (ELISA) was used for the blood meal analysis and sporozoite detection. Five species were identified out of 16,410 anophelines collected. An. gambiae s.s made up approximately 29.2%-36.6% of the population in each zone. All five species acted as vectors for P. falciparum. An. gambiae s.s had the highest sporozoite rate. The most infected mosquitoes were found in the rain forest. More blood meals were taken from bovids, except the savannah forest, where 73.3% were on humans and Human Blood index (HBI) was 57.3%. The Entomological inoculation rate (EIR) was a mean of 13.6 ib/p but was highest in the rainforest zone. This study demonstrates the complex distribution of anophelines and the considerable variations in the intensity of malaria transmission in Nigeria. We highlight the need to consider diverse epidemiological situations when planning countrywide control programmes.

Increased Endophily by the Malaria Vector Anopheles arabiensis in Southern Zambia and Identification of Digested Blood Meals

An increase in Anopheles arabiensis showing endophilic behavior was observed in Macha, Zambia during March 2007. To determine whether this shift in resting behavior was accompanied by a change in feeding preference, an attempt was made to calculate the human blood index. However, only 46.2% of blood meals were successfully identified with existing polymerase chain reaction (PCR) diagnostics. This failure was hypothesized to be caused by the limitations of existing methods that are not capable of identifying host source from anophelines resting for extended time periods. Using an assay we developed that allows for the identification of mammalian host DNA out to 60 hours post-feeding, we were able to successfully determine the host source of 94.3% of recovered blood meals. The data show that, although An. arabiensis in Macha experienced a period of higher endophily, the degree of anthropophily and the sporozoite rate in the population remained comparable to the previous malaria season.

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.

Dynamics of multiple insecticide resistance in the malaria vector Anopheles gambiae in a rice growing area in South-Western Burkina Faso

BackgroundInsecticide resistance of the main malaria vector, Anopheles gambiae, has been reported in south-western Burkina Faso, West Africa. Cross-resistance to DDT and pyrethroids was conferred by alterations at site of action in the sodium channel, the Leu-Phe kdr mutation; resistance to organophosphates and carbamates resulted from a single point mutation in the oxyanion hole of the acetylcholinesterase enzyme designed as ace-1R.MethodsAn entomological survey was carried out during the rainy season of 2005 at Vallée du Kou, a rice growing area in south-western Burkina Faso. At the Vallée du Kou, both insecticide resistance mechanisms have been previously described in the M and S molecular forms of An. gambiae. This survey aimed i) to update the temporal dynamics and the circumsporozoite infection rate of the two molecular forms M and S of An. gambiae ii) to update the frequency of the Leu-Phe kdr mutation within these forms and finally iii) to investigate the occurrence of the ace-1R mutation.Mosquitoes collected by indoor residual collection and by human landing catches were counted and morphologically identified. Species and molecular forms of An. gambiae, ace-1R and Leu-Phe kdr mutations were determined using PCR techniques. The presence of the circumsporozoite protein of Plasmodium falciparum was determined using ELISA.ResultsAnopheles gambiae populations were dominated by the M form. However the S form occurred in relative important proportion towards the end of the rainy season with a maximum peak in October at 51%. Sporozoite rates were similar in both forms. The frequency of the Leu-Phe kdr mutation in the S form reached a fixation level while it is still spreading in the M form. Furthermore, the ace-1R mutation prevailed predominately in the S form and has just started spreading in the M form. The two mutations occurred concomitantly both in M and S populations.ConclusionThese results showed that the Vallée du Kou, a rice growing area formerly occupied mainly by M susceptible populations, is progressively colonized by S resistant populations living in sympatry with the former. As a result, the distribution pattern of insecticide resistance mutations shows the occurrence of both resistance mechanisms concomitantly in the same populations. The impact of multiple resistance mechanisms in M and S populations of An. gambiae on vector control measures against malaria transmission, such as insecticide-treated nets (ITNs) and indoor residual spraying (IRS), in this area is discussed.

Malaria Persistence in Kumaon Foothills of District Nainital, Uttarakhand, India

Clinic data depicted a high incidence of malaria in a forest ecotype in the Kumaon foothills of District Nainital, Uttarakhand, India. A study was therefore conducted to determine the risk factors associated with the transmission of malaria from 2002 to 2004. The man-hour densities of Anopheles culicifacies and An. fluviatilis varied from 2 to 139 and 1 to 69, respectively. The sporozoite rate of 0.24% was recorded in An. culicifacies. Sibling species investigation revealed prevalence of species B (53.8%) and C (46.2%) of An. culicifacies and of species T (100%) of An. fluviatilis complexes. The slide positivity rate and slide falciparum rate were 50.4% and 28.3%, respectively. The infant parasite rate was 42.9% and the enlarged spleen rate among the children was 25.95%. The findings revealed the persistence of malaria in the area mainly due to indigenous transmission through the malaria vector An. culicifacies and the inadequacy of intervention measures. Transmission could be curtailed by indoor residual application of malathion.

Mosquito abundance, bed net coverage and other factors associated with variations in sporozoite infectivity rates in four villages of rural Tanzania

BackgroundEntomological surveys are of great importance in decision-making processes regarding malaria control strategies because they help to identify associations between vector abundance both species-specific ecology and disease intervention factors associated with malaria transmission. Sporozoite infectivity rates, mosquito host blood meal source, bed net coverage and mosquito abundance were assessed in this study.MethodologyA longitudinal survey was conducted in four villages in two regions of Tanzania. Malaria vectors were sampled using the CDC light trap and pyrethrum spray catch methods. In each village, ten paired houses were selected for mosquitoes sampling. Sampling was done in fortnight case and study was undertaken for six months in both Kilimanjaro (Northern Tanzania) and Dodoma (Central Tanzania) regions.ResultsA total of 6,883 mosquitoes were collected including: 5,628 (81.8%) Anopheles arabiensis, 1,100 (15.9%) Culex quinquefasciatus, 89 (1.4%) Anopheles funestus, and 66 (0.9%) Anopheles gambiae s.s. Of the total mosquitoes collected 3,861 were captured by CDC light trap and 3,022 by the pyrethrum spray catch method. The overall light trap: spray catch ratio was 1.3:1. Mosquito densities per room were 96.5 and 75.5 for light trap and pyrethrum spray catch respectively. Mosquito infectivity rates between villages that have high proportion of bed net owners and those without bed nets was significant (P < 0.001) and there was a significant difference in sporozoite rates between households with and without bed nets in these four villages (P < 0.001).ConclusionMalaria remains a major problem in the study areas characterized as low transmission sites. Further studies are required to establish the annual entomological inoculation rates and to observe the annual parasitaemia dynamics in these communities. Outdoor mosquitoes collection should also be considered.

Effect of Rice Cultivation on Malaria Transmission in Central Kenya

A 12-month field study was conducted between April 2004 and March 2005 to determine the association between irrigated rice cultivation and malaria transmission in Mwea, Kenya. Adult mosquitoes were collected indoors twice per month in three villages representing non-irrigated, planned, and unplanned rice agro-ecosystems and screened for blood meal sources and Plasmodium falciparum circumsporozoite proteins. Anopheles arabiensis Patton and An. funestus Giles comprised 98.0% and 1.9%, respectively, of the 39,609 female anophelines collected. Other species including An. pharoensis Theobald, An. maculipalpis Giles, An. pretoriensis Theobald, An. coustani Laveran, and An. rufipes Gough comprised the remaining 0.1%. The density of An. arabiensis was highest in the planned rice village and lowest in the non-irrigated village and that of An. funestus was significantly higher in the non-irrigated village than in irrigated ones. The human blood index (HBI) for An. arabiensis was significantly higher in the non-irrigated village compared with irrigated villages. For An. funestus, the HBI for each village differed significantly from the others, being highest in the non-irrigated village and lowest in the planned rice village. The sporozoite rate and annual entomologic inoculation rate (EIR) for An. arabiensis was 1.1% and 3.0 infective bites per person, respectively with no significant difference among villages. Sporozoite positive An. funestus were detected only in planned rice and non-irrigated villages. Overall, 3.0% of An. funestus samples tested positive for Plasmodium falciparum sporozoites. The annual EIR of 2.21 for this species in the non-irrigated village was significantly higher than 0.08 for the planned rice village. We conclude that at least in Mwea Kenya, irrigated rice cultivation may reduce the risk of malaria transmission by An. funestus but has no effect on malaria transmission by An. arabiensis. The zoophilic tendency of malaria vectors in irrigated areas accounts partly for low malaria transmission rates despite the presence of higher vector densities, highlighting the potential of zooprophylaxis in malaria control.

Abundance, host preferences and infectivity rates of malaria vectors in Badagry Local Government Area of Lagos, Nigeria

The role of anopheles mosquitoes in malaria transmission was investigated in Badagry Local Government Area of Lagos, Nigeria, between July-October 2004. A total of 1,800 female anopheline mosquitoes were collected indoors by human bait and pyrethrum sprays in this coastal rainforest zone. Species identification morphologically and by polymerase chain reaction (PCR) was carried out. ELISA was used to examine P. falciparium sporozoites-infected mosquitoes and their blood meal origin. An.gambiae s.s (39%), An.funestus s.s (16.9%), An.moucheti nigeriensis (15.7%), An. arabiensis (11.7%) and An. melas (10.2%) were identified. Sporozoite rate was 4.2 %, Human blood index (HBI) 67.3% and average monthly Entomological Inoculation Rate (EIR) was 8.4 infective bites per person per month (ib/p/m) by human bait and 5.45 ib/p/m by pyrethrum sprays. This study highlights the complex distribution of different species in the zone. The combined contribution of these mosquito species to malaria transmission in this study highlights the inappropriateness of vector control strategies aimed at one vector. This shows that diverse epidemiological situations have to be taken into account to control malaria. Keywords : Malaria; vectors; Plasmodium falciparium ; transmission dynamics; Nigeria. Nigerian Journal of Parasitology Vol. 27 2006: pp. 41-48

Entomological survey and infection rates of &lt;i&gt;Plasmodium falciparum&lt;/i&gt; and &lt;i&gt;Wuchereria bancrofti&lt;/i&gt; in mosquito populations in the Kainji Lake Area, Nigeria

An entomological study was conducted on vectors of malaria and bancroftian filariasis in Kainji Lake Area of Niger State, Nigeria. A total of 1,545 mosquitoes were collected in this study, and of this, anopheline mosquito represented 72.6% of the total collected while culicine mosquito comprised only 27.4%. Anopheles gambiae complex constituted 60% of the mosquitoes collected. A further identification with PCR showed that An. arabiensis and An. gambiae s.s were 34.5% and 28.5 % (of the mosquitoes collected) respectively. Plasmodium falciparum sporozoites rate of 6.7% for An. arabiensis and 4.6% for An. gambiae s.s were obtained by ELISA. None of the 1,122 anopheline tested was positive for Plasmodium malariae. Dissection of Anopheles, Culex and Aedes species revealed infective larvae of Wuchereria bancrofti only in Anopheles gambiae s.s. with an overall infection rate of 0.5%. These results indicate that the current malaria vector control using pyrethroid treated nets also stand a chance of controlling the vector of bancroftian filariasis in communities where both diseases co-exist. Keywords: mosquito, vector, malaria, filariasis, Kanji, Nigeria. Nigerian Journal of Parasitology Vol. 27 2006: pp.58-61

Malaria vector control by indoor residual insecticide spraying on the tropical island of Bioko, Equatorial Guinea

BackgroundA comprehensive malaria control intervention was initiated in February 2004 on Bioko Island, Equatorial Guinea. This manuscript reports on the continuous entomological monitoring of the indoor residual spray (IRS) programme during the first two years of its implementation.MethodsMosquitoes were captured daily using window traps at 16 sentinel sites and analysed for species identification, sporozoite rates and knockdown resistance (kdr) using polymerase chain reaction (PCR) to assess the efficacy of the vector control initiative from December 2003 to December 2005.ResultsA total of 2,807 and 10,293 Anopheles funestus and Anopheles gambiae s.l. respectively were captured throughout the study period. Both M and S molecular forms of An. gambiae s.s. and Anopheles melas were identified. Prior to the first round of IRS, sporozoite rates were 6.0, 8.3 and 4.0 for An. gambiae s.s., An. melas and An. funestus respectively showing An. melas to be an important vector in areas in which it occurred. After the third spray round, no infective mosquitoes were identified. After the first spray round using a pyrethroid spray the number of An. gambiae s.s. were not reduced due to the presence of the kdr gene but An funestus and An. melas populations declined from 23.5 to 3.1 and 5.3 to 0.8 per trap per 100 nights respectively. After the introduction of a carbamate insecticide in the second round, An. gambiae s.s. reduced from 25.5 to 1.9 per trap per 100 nights and An. funestus and An. melas remained at very low levels. Kdr was found only in the M-form of An. gambiae s.s. with the highest frequency at Punta Europa (85%).ConclusionAll three vectors that were responsible for malaria transmission before the start of the intervention were successfully controlled once an effective insecticide was used.Continuous entomological surveillance including resistance monitoring is of critical importance in any IRS based malaria vector control programme. This paper demonstrates that sufficient resources for such monitoring should be included in any proposal in order to avoid programme failures.

MALARIA VECTOR INCRIMINATION IN THREE RURAL RIVERINE VILLAGES IN THE BRAZILIAN AMAZON

Vector incrimination studies were conducted from April 2003 to February 2005 at three riverine villages 1.5 km to 7.0 km apart, along the Matapi River, Amapa State, Brazil. A total of 113,117 mosquitoes were collected and placed in pools of <or=7 mosquitoes (19,883 pools) and tested for species-specific circumsporozoite protein (CSP) of P. falciparum, P. vivax VK210, and P. vivax VK247 using the enzyme-linked immunoabsorbent assay (ELISA). A subset of 63,330 mosquitoes (12,191 pools) was tested for P. malariae. Anopheles darlingi and An. marajoara had the highest proportion of circumsporozoite protein positives for human malaria parasites compared with An. nuneztovari, An. triannulatus, and An. intermedius. Anopheles darlingi and An. marajoara had the highest entomological inoculation rates (EIR) and were considered to be the most important malaria vectors in the study. Anopheles nuneztovari was also an important vector. Differences in entomological inoculation rates were more dependent on mosquito abundance than on sporozoite rates.

Blood‐feeding behaviour of the malarial mosquitoAnopheles arabiensis: implications for vector control

Feeding behaviour of the malaria vector Anopheles arabiensis Patton (Diptera: Culicidae) was monitored for 12 months (March 2003-February 2004) in the Konso District of southern Ethiopia (5 degrees 15'N, 37 degrees 28'E). More than 45 000 An. arabiensis females were collected by host-baited sampling methods (light-traps, human landing catches, cattle-baited traps) and from resting sites (huts and pit shelters). In the village of Fuchucha, where the ratio of cattle : humans was 0.6 : 1, 51% of outdoor-resting mosquitoes and 66% of those collected indoors had fed on humans, human baits outdoors caught > 2.5 times more mosquitoes than those indoors and the mean catch of mosquitoes from pit shelters was about five times that from huts. Overall, the vast majority of feeding and resting occurred outdoors. In the cattle camps of Konso, where humans slept outdoors close to their cattle, approximately 46% of resting mosquitoes collected outdoors had fed on humans despite the high cattle : human ratio (17 : 1). In both places, relatively high proportions of bloodmeals were mixed cow + human: 22-25% at Fuchucha and 37% in the cattle camps. Anthropophily was also gauged experimentally by comparing the numbers of mosquitoes caught in odour-baited entry traps baited with either human or cattle odour. The human-baited trap caught about five times as many mosquitoes as the cattle-baited one. Notwithstanding the potential pitfalls of using standard sampling devices to analyse mosquito behaviour, the results suggest that the An. arabiensis population is inherently anthropophagic, but this is counterbalanced by exophagic and postprandial exophilic tendencies. Consequently, the population feeds sufficiently on humans to transmit malaria (sporozoite rates: 0.3% for Plasmodium falciparum and 0.5% for P. vivax, by detection of circumsporozoite antigen) but also takes a high proportion of meals from non-human hosts, with 59-91% of resting mosquitoes containing blood from cattle. Hence, classical zooprophylaxis is unlikely to have a significant impact on the malaria vectorial capacity of An. arabiensis in Konso, whereas treating cattle with insecticide might do.

The relationship between malaria parasitaemia and availability of healthcare facility in Mpwapwa District, central Tanzania

A study was carried out in six villages located at different altitudes in Mpwapwa district of central Tanzania to determine malaria parasitaemia and transmission levels in villages with or without health care facilities. A total of 1119 schoolchildren (age = 5.9-12.3 years) were examined for malaria parasitaemia. Plasmodiumfalciparum was the predominant malaria species accounting for 92.8% of all species. The average malaria prevalence rate among schoolchildren was 25.8% (range 1.5-53.8%). The geometric mean parasite densities for P.falciparum was 361 (N = 286). Higher malaria prevalence was observed in villages at lower (< 1000 m) than at intermediate (1000-1500m) or higher (> 1500m) altitudes. Schoolchildren in areas with health care facilities were less at risk of acquiring malaria by 33.4% as compared with those living in areas without health facilities. Mean packed cell volume in schoolchildren was 38.5% (range = 35.2-41.0%). Splenomegaly was observed in 18.1% (0-40.2%) of the schoolchildren examined and it was higher among those in villages without health care facilities. Anopheles gambiae sensu lato was the only malaria vector found in the district and was found in all villages and at all altitudes. Sporozoite rate in An. gambiae s.l. ranged from 0-10.5%, with the lowland villages recording the highest rates. This study indicates that altitude and geographical accessibility to healthcare service are important determinants of malaria infection among rural communities in Tanzania.

Malaria control using indoor residual spraying and larvivorous fish: a case study in Betul, central India

To assess the impact of intensified malaria control interventions in an ethnic minority community in Betul using existing tools. Two rounds of indoor residual spraying with synthetic pyrethroid insecticide were applied and larvivorous fish introduced, followed by intensive surveillance for early detection of Plasmodium falciparum with rapid diagnostic tests and prompt treatment with sulphadoxine pyrimethamine. Pre-intervention surveys revealed a very high fever rate in the community in all age groups with a slide positivity rate of >50% with >90%P. falciparum. The post-intervention phase showed a sharp steady decline in number of malaria cases (beta 0.972; P < 0.0001, 95% CI 0.35-0.47). Monitoring of entomological results revealed a significant decline in both Anopheles species and An. culicifacies (P < 0.0001). A combination of indoor residual spraying and early detection and prompt treatment complemented by rapid diagnostic tests and larvivorous fishes successfully brought malaria under control. These approaches could be applied in other regions of different endemicity to control malaria in India.

Sampling techniques for adult Afrotropical malaria vectors and their reliability in the estimation of entomological inoculation rate

Various entomological indicators and sampling techniques are used to monitor and evaluate the impact of many vector control interventions. A number of methods have been used in sampling mosquitoes for the purpose of estimating the entomological inoculation rate (EIR) and each is subject to some bias or shortcomings. It was the aim of this paper to critically evaluate the most common mosquito sampling techniques in relation to their reliability in the estimation of EIR. The techniques include man-landing, light trap, light trap/bednet combination and odour-baited traps. Although man-landing technique is the most reliable, it however, expose the catcher to mosquito-borne infections. On the other hand, light traps have been found to capture mosquitoes with higher sporozoite rates as compared to those from human bait catch thus leading to an overestimation of EIR. From an epidemiological point of view, the use of light-trap-bed net combination is an approach that is more meaningful than using light trap alone because, a light trap functions more efficiently when placed near the normal flight paths of mosquitoes such as inside huts or under the eaves. Unfortunately, it has been shown that estimates of EIR are influenced by trap position, hence affecting the number caught and the sporozoite rates. A variety of bednets have been used to sample mosquitoes attracted to man. Studies have shown that bednet traps normally catch fewer mosquitoes than do human baits outside them. Although the collections by indoor resting technique give a good estimate of the mean house density in a given area, they may not necessarily give a good estimate of EIR. Thus the development of improved sampling systems based on an improved understanding of host-oriented behaviour is needed. Moreover, there is need to standardise all the sampling techniques in use to enable us make valid comparisons between various studies done by different people and in different areas. In this article, the inherent limitations of conventional mosquito sampling techniques when used in estimating the EIR are discussed.

Are insecticide-treated bednets more protective against Plasmodium falciparum than Plasmodium vivax- infected mosquitoes?

BackgroundThe outcomes of insecticide-treated bednet (ITN) interventions for malaria control in Papua New Guinea tend to suggest a differential protective effect against Plasmodium falciparum and Plasmodium vivax. Little is known about the impact of ITNs on the relative abundance of mosquitoes infected with either P. falciparum or P. vivax. This paper describes the biting cycle of P. falciparum and P. vivax-infected mosquitoes and the impact of an ITN intervention on the proportion of mosquitoes infected with either parasite species.MethodsEntomological investigations were performed in East Sepik (ESP) and New Ireland Provinces (NIP) of PNG. Mosquitoes were collected using the all-night (18:00 - 06:00) landing catch and CDC light-trap methods and species specific malaria sporozoite rates were determined by ELISA.Results and discussionThe distribution of sporozoite positive mosquitoes in three four-hour periods (18:00-22:00, 22:00-02:00 & 02:00-06:00) showed that a higher proportion of P. vivax-infected mosquitoes were biting before people retired to bed under the protection of bednets. In the intervention village, the 308 mosquitoes collected before ITNs were introduced included eight (2.0%) P. falciparum-positive and four (1.0%) P. vivax-positive specimens, giving a parasite ratio of 2:1. The sporozoite rate determined from 908 mosquitoes caught after ITNs were introduced showed a significant decrease for P. falciparum (0.7%) and a slight increase for P. vivax (1.3%), resulting in a post intervention parasite ratio of 1:2. In the East Sepik Province, where ITNs were not used, P. falciparum remained the dominant species in 12 monthly mosquito collections and monthly P. falciparum:P. vivax formula varied from 8:1 to 1.2:1.ConclusionThese findings suggest that people sleeping under treated bednets may be more exposed to P. vivax than P. falciparum-infected mosquitoes before going to sleep under the protection of bednets. This difference in the biting behaviour of mosquitoes infected with different malaria parasites may partly explain the change in the P. falciparum:P. vivax formula after the introduction of ITNs.