Sporozoite Rate Research Articles

THE USE OF NESTED-PCR TO DETECT THE PRESENCE OF PLASMODIUM IN ANOPHELES ARABIENSIS IN JAZAN REGION, SAUDI ARABIA

The present study was carried out in 26 viJlages at two Governates (Al-Khobah, and Haroob) in Jazan Region in Southwest Saudi Arabia to identify and detect the presence of Plasmodium in Anopheles arabiensis using nested-PCR technique. An. Arabiensis was identified by PCR and it was the predominant Anopheles mosquito in all the collection sites. A total of 257 An. Arabiensis females were collected and two samples from two villages (Almuatan and Alsabkha) out of 107 (1.87%) female mosquitoes from Haroob Governorate were found positive for the sporozoites of Plasmodium falciparum. Similarly, 3 out of 150 (2%) female mosquito samples from Um-alkhameir, AL-Khobah Governorate, were also found positive. Around fourfold increase of the sporozoite rate (from 0.61 to 2.0%) in An. arabiensis in Al-Khobah Governorate has been observed compared to the previous study of 2007-2008. The wide spread of An. arabiensis in Jazan region with >90% of the malaria cases caused by P. falciparum, along with infectivity rate ranges between 1.87 to 2.0% for P. falciparum in Al-Khobah and Haroob Governorates, suggests that P. falciparumn is the most predominant malaria parasite and An. Arabiensis is a very efficient malaria vector in the region. It also suggests more in-depth researches on the ecology, behavior, and control of An. Arabiensis to promote area-specific control programs.

Entomological Surveillance of Malaria Vectors in Saumlaki, Maluku Tenggara Barat Regency, Maluku Province

The research aims to determine the prevalence of malaria and Anopheles spp using bio-ecology surveillance in Alusi and Waturu community health centers in Maluku Tenggara Barat Regency. The study was conducted in March-April 2015 with cross sectional design. In this research, we performed mass blood survey on 489 participants in the Kilmasa village and 434 participants in Waturu village. We also performed entomology surveillances, i.e. larval density, catching Anopheles spp, temperature, humidity, and salinity. To confirm malaria vectors, we used enzyme linked immunoabsorbent assay (ELISA) techniques. The data analyzed descriptively. The results of the study showed proportion 0.20% malaria morbidity in Kilmasa village and 0.23% in Waturu village. Anopheles flavirostris and An. barbirostris group were likely to bite a human outside and inside the house and peaked at 11.00 pm-12.00 pm. The parous rate of An. flavirostris and An. barbirostris was 46% and 26%, respectively. Human blood index of An.flavirostris and An. barbirostris was 33.3% and by 70%, respectively. Anopheles flavirostri and An. barbirostris were malaria vectors with sporozoite rate 0.38% and 12.5%, respectively.

Beyond the entomological inoculation rate: characterizing multiple blood feeding behavior and Plasmodium falciparum multiplicity of infection in Anopheles mosquitoes in northern Zambia

BackgroundA commonly used measure of malaria transmission intensity is the entomological inoculation rate (EIR), defined as the product of the human biting rate (HBR) and sporozoite infection rate (SIR). The EIR excludes molecular parameters that may influence vector control and surveillance strategies. The purpose of this study was to investigate Anopheles multiple blood feeding behavior (MBF) and Plasmodium falciparum multiplicity of infection (MOI) within the mosquito host in Nchelenge District, northern Zambia. Mosquitoes were collected from light traps and pyrethroid spray catch in Nchelenge in the 2013 wet season. All anophelines were tested for blood meal host, P. falciparum, and MOI using PCR. Circumsporozoite (CSP) ELISA and microsatellite analysis were performed to detect parasites in the mosquito and MBF, respectively. Statistical analyses used regression models to assess MBF and MOI and exact binomial test for human sex bias. Both MBF and MOI can enhance our understanding of malaria transmission dynamics beyond what is currently understood through conventional EIR estimates alone.ResultsThe dominant malaria vectors collected in Nchelenge were Anopheles funestus (sensu stricto) and An. gambiae (s.s.) The EIRs of An. funestus (s.s.) and An. gambiae (s.s.) were 39.6 infectious bites/person/6 months (ib/p/6mo) and 5.9 ib/p/6mo, respectively, and took multiple human blood meals at high rates, 23.2 and 25.7% respectively. There was no bias in human host sex preference in the blood meals. The SIR was further characterized for parasite genetic diversity. The overall P. falciparum MOI was 6.4 in infected vectors, exceeding previously reported average MOIs in humans in Africa.ConclusionsBoth Anopheles MBF rates and P. falciparum MOI in Nchelenge were among some of the highest reported in sub-Saharan Africa. The results suggest an underestimation of the EIR and large numbers of circulating parasite clones. Together, the results describe important molecular aspects of transmission excluded from the traditional EIR measurement. These elements may provide more sensitive measures with which to assess changes in transmission intensity and risk in vector and parasite surveillance programs.

Spatio-temporal heterogeneity of malaria vectors in northern Zambia: implications for vector control.

BackgroundDespite large reductions in malaria burden across Zambia, some regions continue to experience extremely high malaria transmission. In Nchelenge District, Luapula Province, northern Zambia, almost half the human population carries parasites. Intervention coverage has increased substantially over the past decade, but comprehensive district-wide entomological studies to guide delivery of vector control measures are lacking. This study describes the bionomics and spatio-temporal patterns of malaria vectors in Nchelenge over a two and a half year period, investigates what household factors are associated with high vector densities and determines why vector control may not have been effective in the past to better guide future control efforts.MethodsBetween April 2012 and September 2014, twenty-seven households from across Nchelenge District were randomly selected for monthly light trap collections of mosquitoes. Anopheline mosquitoes were identified morphologically and molecularly to species. Foraging rates were estimated and sporozoite rates were determined by circumsporozoite ELISAs to calculate annual entomological inoculation rates. Blood feeding rates and host preference were determined by PCR. Zero-inflated negative binomial models measured environmental and household factors associated with mosquito abundance at study households such as season, proximity to the lake, and use of vector control measures.ResultsThe dominant species in Nchelenge was An. funestus (s.s.) with An. gambiae (s.s.) as a secondary vector. Both vectors were found together in large numbers across the district and the combined EIRs of the two vectors exceeded 80 infectious bites per person per annum. An. funestus household densities increased in the dry season whilst An. gambiae surged during the rains. Presence of insecticide treated nets (ITNs) and closed eaves in the houses were found to be associated with fewer numbers of An. gambiae but not An. funestus. There was no association with indoor residual spraying (IRS).ConclusionIn Nchelenge, the co-existence of two highly anthropophagic vectors, present throughout the year, is likely to be driving the high malaria transmission evident in the district. The vectors here have been shown to be highly resistant to pyrethroids used for IRS during the study. Vector control interventions in this area would have to be multifaceted and district-wide for effective control of malaria.Electronic supplementary materialThe online version of this article (doi:10.1186/s13071-016-1786-9) contains supplementary material, which is available to authorized users.

Vector Competence of Anopheles kleini and Anopheles sinensis (Diptera: Culicidae) From the Republic of Korea to Vivax Malaria-Infected Blood From Patients From Thailand.

In total, 1,300 each of Anopheles kleini Rueda and Anopheles sinensis Wiedemann sensu stricto (s.s.) females (colonized from the Republic of Korea) and Anopheles dirus Peyton & Harrison (Thai strain) were allowed to feed on blood from Thai malaria patients naturally infected with Plasmodium vivax The overall oocyst infection rates for An. dirus, An. kleini, and An. sinensis s.s. were 77.4, 46.1, and 45.9%, respectively. The mean number of oocysts was significantly higher for An. dirus (82.7) compared with An. kleini (6.1) and An. sinensis s.s. (8.6), whereas the mean number of oocysts for An. kleini and An. sinensis s.s. was similar. The overall sporozoite infection rates for An. dirus, An. kleini, and An. sinensis s.s. dissected on days 14-15, 21, and 28 days post-feed were significantly higher for An. dirus (90.0%) than An. kleini (5.4%), whereas An. kleini sporozoite rates were significantly higher than An. sinensis s.s. (<0.1%). The overall sporozoite indices for positive females with +3 (100-1,000 sporozoites) and +4 (>1,000 sporozoites) salivary gland indices were significantly higher for An. dirus (85.7%), compared with An. kleini (47.1%). Only one An. sinensis s.s. had sporozoites (+2; >10-100 sporozoites). These results indicate that An. kleini is a competent vector of vivax malaria. Although An. sinensis s.s. develops relatively high numbers of oocysts, it is considered a very poor vector of vivax malaria due to a salivary gland barrier.

Kapasitas Vektor dan Laju Inokulasi Entomologis Anopheles vagus dari Wilayah Endemis Malaria di Provinsi Banten

High malaria cases in Banten Province remains in two districts, Pandeglang and Lebak. Yet, the knowledge and information about vector capacity in this region is still unclear. Our study aimed to determine the vector capacity and entomological inoculation rate of Anopheles vagus in endemic regions in Banten Province. A cross-sectional survey has been conducted for June – October 2014 in Lebak and Pandeglang district in 2014. The data of mosquito was collected through human landing collection, ovarian dissection, ELISA ,and PCRbased to determine the man-biting rate (MBR), longevity, vectorial capacity, and entomological inoculation rate (EIR) of An.vagus. As a result, we found 11 species of Anopheline mosquito: An. aconitus, An. annularis, An. barbirostris, An. flavirostris, An. indefinitus, An. kochi, An. maculatus, An. minimus, An. sundaicus, An. tesselatus, and An. vagus. The predominantly species found in Pandeglang is An. vagus with peak density at 3.00-4:00 am (MBR = 2.07individual per person per night). Meanwhile, in Lebak dominated by An. sundaicus with peak densities starting at 10.00 pm (MBR 4.1 individual per person per night), while An. vagus as 24Vektora Volume 8 Nomor 1, Juni 2016: 23 - 30the second dominant species. The sporozoites rate of 1.02%, daily life expectancy = 0.244, longevity = 4.093 days; and Human Blood Index 0.1, then the vector capacity of An. vagus at 0.281 and entomological inoculation rate = 0.021 (~ 7 infective bites/person/ year), respectively. Our study concluded that malaria transmission may still occur in this two areas.

Circumsporozoite protein rates, blood-feeding pattern and frequency of knockdown resistance mutations in Anopheles spp. in two ecological zones of Mauritania.

BackgroundMosquitoes belonging to Anopheles gambiae species complex are the main malaria vector in Mauritania but data on their vector capacities, feeding habits and insecticide susceptibility are still scanty. The objectives of this study were to fill this gap.MethodsAdult Anopheles spp. mosquitoes were collected using pyrethrum spray catch method from two ecological zones of Mauritania: Nouakchott (Saharan zone) and Hodh Elgharbi region (Sahelian zone). Circumsporozoite proteins (CSP) for P. falciparum, P. vivax VK210 and P. vivax VK247 were detected by enzyme-linked immunosorbent assay (ELISA) from the female anopheline mosquitoes. To confirm CSP-ELISA results, polymerase chain reaction (PCR) was also performed. Blood meal identification was performed in all engorged females by partial sequencing of the mitochondrial cytochrome b gene. Molecular assessments of pyrethroid knockdown resistance (kdr) and insensitive acetylcholinesterase resistance (ace-1) were conducted.ResultsIn Nouakchott, the only species of Anopheles identified during the survey was Anopheles arabiensis (356 specimens). In Hodh Elgharbi, 1016 specimens of Anopheles were collected, including 578 (56.9 %) Anopheles rufipes, 410 (40.35 %) An. arabiensis, 20 (1.96 %) An. gambiae, 5 (0.5 %) An. pharoensis and 3 (0.3 %) An. funestus. Three of 186 female An. arabiensis collected in Nouakchott and tested by ELISA were found positive for Plasmodium vivax VK210, corresponding to a sporozoite rate of 1.6 %; however PCR confirmed infection by P. vivax sporozoite in only one of these. In Hodh Elgharbi, no mosquito was found positive for Plasmodium spp. infection. There was a statistically significant difference in the percentage of human blood-fed Anopheles spp. between Nouakchott (58.7 %, 47 of 80 blood-engorged An. arabiensis females) and Hodh Elgharbi (11.1 %, 2 of 18 blood-engorged mosquitoes). Analysis of the kdr polymorphisms showed 48.2 % (70/145) of East African kdr mutation (L1014S) in Nouakchott compared to 10 % (4/40) in Hodh Elgharbi region (P < 0.001). Nevertheless, West African kdr mutation (L1014F) was found only in An. gambiae populations (4/40, 10 %) from Hodh Elgharbi region. No ace-1 mutation was found in mosquito specimens from the two study zones.ConclusionsOverall, this study confirmed the autochthonous P. vivax malaria transmission in Nouakchott, involving An. arabiensis as the main vector. It also described for the first time the absence of ace-1 mutation, the co-occurrence of both West and East African kdr mutation in An. gambiae in Mauritania, and highlighted the regional variations in the prevalence and type of kdr mutations.

Partitioning the contributions of alternative malaria vector species.

BackgroundIn many locations malaria is transmitted by more than one vector species. Some vector control interventions, in particular those using genetic approaches, are likely to be targeted against a single species or species complex, at least initially, and it would therefore be useful to be able to predict the epidemiological impact of controlling a single species when multiple vector species are present.MethodsTo address this issue, the classical Ross-McDonald model of malaria epidemiology is expanded to account for multiple vector species, giving expressions for the equilibrium prevalence, sporozoite rates and reproductive number. These allow one to predict when control of just one vector species will lead to elimination of the disease. Application of the model is illustrated using published data from a particularly extensive entomological and epidemiological survey before the rollout of bed nets in eastern Kenya, where Anopheles gambiaes.l. and An. funestus were vectors.ResultsMeta-analysis indicates that sporozoite rates were 38 % higher in An. gambiaes.l. than in An. funestus, and, according to the model, this difference could be due to An. gambiae s.l. having a higher frequency of feeding on humans, a higher human-to-mosquito transmission rate, a lower adult mortality rate, and/or a shorter incubation period. Further calculations suggest that An. gambiaes.l. would have been sufficient to maintain transmission by itself throughout the region, whereas An. funestus would not have been able to support transmission by itself in Malindi District.ConclusionsPartitioning the contributions of different vector species may allow us to predict whether malaria will persist after targeted vector control.

Characteristics of asymptomatic Plasmodium spp. parasitaemia in Kwahu-Mpraeso, a malaria endemic mountainous district in Ghana, West Africa.

BackgroundMalaria control efforts in Ghana have reduced the countrywide average malaria prevalence from 71 % in 2000 to about 51 % in 2012; however, its main focus is on symptomatic malaria. If further progress is to be made, parasite reservoirs in asymptomatic carriers need to be moved into focus. This study profiles asymptomatic Plasmodium spp. parasitaemia amongst residents of mountainous Kwahu-Mpraeso in the Eastern region of Ghana.MethodsA cross-sectional study of 360 residents was carried out from October to December 2013. This included recording demographics, malaria testing of asymptomatic residents, and gathering of their malaria history. Assessment of malaria transmission was done with molecular identification of vectors, determination of sporozoite rate, insecticide resistance status and biting pattern. Univariate and multivariate analysis were used to establish risk determinants.ResultsIn Mpraeso, in the Kwahu highland of Eastern Region, children were at higher risk of asymptomatic parasitaemia, thereby contributing to the parasite reservoir and hence sustained malaria transmission. As well, findings suggested Hb AC genotype influenced susceptibility to asymptomatic malaria with 8.03-fold increase in odds (univariate) and 11.92-fold higher odds (multivariate) than the normal Hb AA. The mosquito vector predominant in the area was Anopheles gambiae sensu stricto of the homozygous pyrethroid resistant form (RR); with biting mainly occurring indoors.ConclusionFor an effective malaria control in this area, interventions should be formulated and implemented to target asymptomatic parasite reservoirs; especially in children and people with Hb AC. The dominant vector species An. gambiae s.s. and its feeding patterns of biting indoors should also be considered.

Malaria case in Madagascar, probable implication of a new vector, Anopheles coustani.

BackgroundIndoor spraying of insecticides and the use of insecticide-treated bed nets are key strategies for national malaria vector control in the central highlands of Madagascar. During the year 2013, malaria outbreaks were reported by the National Malaria Control Programme in the highlands, including the district of Ankazobe.MethodsEntomological trapping was carried out in April and May 2013 and in March 2014, using human landing catches, collection of mosquitoes resting in stables and in houses by oral aspirators, and Centers for Disease Control and Prevention light traps. Detection of Plasmodium in mosquitoes was carried out on head and thorax of anopheline females by ELISA, CSP and PCR (Plasmodium falciparum, Plasmodium malariae, Plasmodium vivax, or Plasmodium ovale). Human biting rate (HBR), sporozoite index and entomological infection rate (EIR) were calculated for Anopheles funestus, Anopheles arabiensis,Anopheles mascarensis, and Anopheles coustani.ResultsIn Ankazobe district, the presence of malaria vectors such as An. funestus, An. arabiensis and An. mascarensis was confirmed, and a new and abundant potential vector, An. coustani was detected. Indeed, one individual of An. funestus and two An. coustani were detected positive with P. falciparum while one An. mascarensis and four An. coustani were positive with P. vivax. For An. coustani, in March 2014, the EIR varied from 0.01 infectious bites/person/month (ipm) outdoors to 0.11 ipm indoors. For An. funestus, in April 2013, the EIR was 0.13 ipm. The highest HBR value was observed for An. coustani, 86.13 ipm outdoors. The highest sporozoite rate was also for An. coustani, 9.5 % of An. coustani caught in stable was sporozoite positive.ConclusionThe implication of An. coustani in malaria transmission was not previously mentioned in Madagascar. Its very high abundance and the detection of Plasmodium coupled with an opportunistic feeding behaviour in villages with malaria cases supports its role in malaria transmission in Madagascar.

Sporogony and sporozoite rates of avian malaria parasites in wild Culex pipiens pallens and C. inatomii in Japan.

BackgroundMalaria infection in mosquitoes is traditionally detected by microscopic examination for Plasmodium oocysts and sporozoites. Although PCR is now widely used, the presence of parasite DNA in a mosquito does not prove that sporogony is achieved. Thus, detection of sporozoites by microscopy is still required to definitively identify vector mosquitoes. The aim of this study was to confirm sporogony of avian Plasmodium spp. in Culex pipiens pallens and C. inatomii caught from the wild.FindingsMosquitoes collected at two sites in Japan were dissected and examined by microscopy for Plasmodium oocysts and sporozoites. DNA was extracted from the midgut and salivary gland of infected mosquitoes, and the infecting Plasmodium species was identified by sequencing 478 bp of cytochrome b. Oocysts, or both oocysts and sporozoites, were found in 3.94 and 0.46 % of C. p. pallens and C. inatomii, respectively. Four (CXPIP09, GRW4, GRW11 and SGS1) and three cytochrome b lineages (CXINA01, CXINA02 and CXQUI01) were confirmed to achieve sporogony in C. p. pallens and C. inatomii, respectively. One mosquito each of C. p. pallens and C. inatomii was co-infected with two different Plasmodium lineages.ConclusionsThese findings demonstrate that C. p. pallens and C. inatomii are natural vectors of four and three lineages of avian Plasmodium spp., respectively. The data indicate that a systematic procedure combining microscopy and PCR is a feasible and reliable approach to identify natural vectors of wildlife malaria.

A shift in resting habitat and feeding behavior of Anopheles fluviatilis sibling species in the Keonjhar district of Odisha, India.

Anopheles fluviatilis exists as a complex of sibling species S, T, U and V exhibiting distinct variations. Sibling species S is considered as the main vector and anthropogenic whereas T, U and V are zoophagic non-vectors. This study was performed in a forested village of Keonjhar district, Odisha to identify the status of An. fluviatilis sibling species. Mosquito collections were made from cattle sheds (CS), human dwellings (HD) and mixed dwellings (MD) from June 2012 to May 2013. The proportion of An. fluviatilis collected from different habitats was compared with An. culicifacies. PCR assays were conducted to reveal their sibling species composition, host preference and sporozoite rate. Anopheles fluviatilis was the dominant species followed by An. culicifacies. The relative proportion of collection was high in MD and HD for An. fluviatilis and An. culicifacies respectively. PCR assay confirmed 9.4% S and 75.5% T. Mean collection of sibling species T and S were significantly high in MD and HD. Human blood index (HBI) of 0.88 and 0.61 was confirmed for sibling species S and T respectively with 13% sporozoite rate for S. High density of the sibling T was found in the study site with a shift in resting habitat and blood feeding preference. GenBank submissions: KJ451071.1, KJ451072.1, KJ451073.1, KJ451074.1, KJ451432.1, KJ451433.1, KJ451434.1, KJ451435.1, KJ451428.1, KJ451429.1, KJ451430.1, KJ451431.1.

Estimating malaria transmission from humans to mosquitoes in a noisy landscape.

A basic quantitative understanding of malaria transmission requires measuring the probability a mosquito becomes infected after feeding on a human. Parasite prevalence in mosquitoes is highly age-dependent, and the unknown age-structure of fluctuating mosquito populations impedes estimation. Here, we simulate mosquito infection dynamics, where mosquito recruitment is modelled seasonally with fractional Brownian noise, and we develop methods for estimating mosquito infection rates. We find that noise introduces bias, but the magnitude of the bias depends on the ‘colour' of the noise. Some of these problems can be overcome by increasing the sampling frequency, but estimates of transmission rates (and estimated reductions in transmission) are most accurate and precise if they combine parity, oocyst rates and sporozoite rates. These studies provide a basis for evaluating the adequacy of various entomological sampling procedures for measuring malaria parasite transmission from humans to mosquitoes and for evaluating the direct transmission-blocking effects of a vaccine.

Early biting of the Anopheles gambiae s.s. and its challenges to vector control using insecticide treated nets in western Kenya highlands

Long term use of insecticides in malaria vector control has been shown to alter the behavior of vectors. Such behavioral shifts have the potential of undermining the effectiveness of insecticide-based control interventions. The effects of insecticide treated nets (ITNs) use on the composition, biting/feeding and sporozoite rates of Anopheles gambiae s.l. mosquitoes in Musilongo village, Vihiga County of western Kenya highlands were investigated. Adult mosquitoes were collected in selected sleeping spaces inside six randomly selected houses using miniature Centre for Disease Control and Prevention (CDC) light traps. Mosquito sampling in each house was conducted twice every week for 16 consecutive months (May 2010–August 2012). At each sampling a single trap was set in the selected space inside each house such that it collected mosquitoes alternatively from 18:00 to 21:00h and 21:00 to 06:00h every week. All collected mosquitoes were morphologically identified. Female Anopheles mosquitoes were classified according to their physiological status as unfed, fed, partially gravid and gravid, sorted and counted. Members of the A. gambiae complex were identified using a Polymerase chain reaction (PCR) method. Enzyme-linked-immunosorbent assay (ELISA) was used to determine blood meal sources and Plasmodium infection rates in A. gambiae s.l. mosquitoes. Blood meal tests were conducted on DNA extracted from gut contents of blood fed A. gambiae s.l. The head and thorax section of dried samples of A. gambiae s.l. were used in testing for the presence of Plasmodium falciparum (Pf) sporozoites. Overall, 735 adult female Anopheles comprising 708 [96.3%] A. gambiae s.l. and 27 [3.7%] Anopheles funestus mosquitoes were collected. A. gambiae s.l. population collected comprised, 615 [86.9%] unfed and 38 [5.4%] fed adult mosquitoes. The rest were either partially or fully gravid. The proportion of A. gambiae s.l. biting indoors within 18:00–21:00h was 15.8% (103/653) at a rate of 3.2bites per person per hour compared to 84.2% biting from 21:00-06:00h at a rate of 3.8 bites/per/h. An estimated 97.7% A. gambiae ss and 2.3% A. arabiensis constituted the indoor biting A. gambiae s.l. The population of An. gambiae s.l. biting from 18:00 to 21:00h had a Plasmodium faciparum (pf) sporozoite rate of 3.8% compared to 3.5% observed in populations biting within 21:00-06:00h. Human blood constituted 89% of An. gambiae s.l. blood meal sources. The risk of malaria transmission from 21:00 to 06:00h was approximately 5 fold the risk within 18:00–21:00h. Majority of the infective female A. gambiae s.l. adults were biting deep into the night than in the early hours of the night. Humans remain the preferred source of blood meal for A. gambiae s.s. the dominant malaria vector in the highlands. ITNs remain a fundamental control intervention against malaria transmission since female blood seekers were more during bed time than pre-bed time. Advocacy on enhanced net availability, integrity and usage in Kenyan highlands can reduce Pf transmission. Additional complementary interventions are required to control the biting and parasite transmission encountered before bed-time.

Human biting activity, spatial-temporal distribution and malaria vector role of Anopheles calderoni in the southwest of Colombia.

BackgroundAnopheles calderoni was first recognized in Colombia in 2010 as this species had been misidentified as Anopheles punctimacula due to morphological similarities. An. calderoni is considered a malaria vector in Peru and has been found naturally infected with Plasmodium falciparum in Colombia. However, its biting behaviour, population dynamics and epidemiological importance have not been well described for Colombia.MethodsTo assess the contribution of An. calderoni to malaria transmission and its human biting behaviour and spatial/temporal distribution in the southwest of Colombia, human landing catches (HLC) and larval collections were carried out in a cross-sectional, entomological study in 22 localities between 2011 and 2012, and a longitudinal study was performed in the Boca de Prieta locality in Olaya Herrera municipality between July 2012 and June 2013. All mosquitoes determined as An. calderoni were tested by ELISA to establish infection with Plasmodium spp.ResultsLarvae of An. calderoni were found in four localities in 12 out of 244 breeding sites inspected. An. calderoni adults were collected in 14 out of 22 localities during the cross-sectional study and represented 41.3% (459 of 1,111) of the collected adult specimens. Other species found were Anopheles albimanus (54.7%), Anopheles apicimacula (2.1%), Anopheles neivai (1.7%), and Anopheles argyritarsis (0.2%). In the localities that reported the highest malaria Annual Parasite Index (>10/1,000 inhabitants) during the year of sampling, An. calderoni was the predominant species (>90% of the specimens collected). In the longitudinal study, 1,528 An. calderoni were collected by HLC with highest biting rates in February, May and June 2013, periods of high precipitation. In general, the species showed a preference to bite outdoors (p < 0.001). In Boca de Prieta, two specimens of An. calderoni were ELISA positive for Plasmodium circumsporozoite protein: one for P. falciparum and one for Plasmodium vivax VK-210. This represents an overall sporozoite rate of 0.1% and an annual entomological inoculation rate of 2.84 infective bites/human/year.ConclusionsThis study shows that An. calderoni is a primary malaria vector in the southwest of Colombia. Its observed preference for outdoor biting is a major challenge for malaria control.

Surveillance of malaria vector population density and biting behaviour in western Kenya.

BackgroundMalaria is a great public health burden and Africa suffers the largest share of malaria-attributed deaths. Despite control efforts targeting indoor malaria transmission, such as insecticide-treated bed nets (ITNs) and deployment of indoor residual spraying, transmission of the parasite in western Kenya is still maintained. This study was carried out to determine the impact of ITNs on indoor vector densities and biting behaviour in western Kenya.MethodsIndoor collection of adult mosquitoes was done monthly in six study sites in western Kenya using pyrethrum spray collections from 2012 to 2014. The rotator trap collections were done in July–August in 2013 and May–June in 2014. Mosquitoes were collected every 2 h between 18.00 and 08.00 h. Human behaviour study was conducted via questionnaire surveys. Species within Anopheles gambiae complex was differentiated by PCR and sporozoite infectivity was determined by ELISA. Species distribution was determined and bed net coverage in the study sites was recorded.ResultsDuring the study a total of 5,469 mosquito vectors were collected from both PSC and Rotator traps comprising 3,181 (58.2%) Anopheles gambiae and 2,288 (41.8%) Anopheles funestus. Compared to all the study sites, Rae had the highest density of An. gambiae with a mean of 1.2 (P < 0.001) while Kombewa had the highest density of An. funestus with a mean of 1.08 (P < 0.001). Marani had the lowest density of vectors with 0.06 An. gambiae and 0.17 An. funestus (P < 0.001). Among the 700 PCR confirmed An. gambiaes.l. individuals, An. gambiaes.s. accounted for 49% and An. arabiensis 51%. Over 50% of the study population stayed outdoors between 18.00 and 20.00 and 06.00 and 08.00 which was the time when highest densities of blood fed vectors were collected. Anopheles gambies.s. was the main malaria parasite vector in the highland sites and An. arabiensis in the lowland sites. Bed net ownership in 2012 averaged 87% across the study sites.ConclusionsThis study suggests that mass distribution of ITNs has had a significant impact on vector densities, species distribution and sporozoite rate. However, shift of biting time poses significant threats to the current malaria vector control strategies which heavily rely on indoor controls.

Relative Abundance and Plasmodium Infection Rates of Malaria Vectors in and around Jabalpur, a Malaria Endemic Region in Madhya Pradesh State, Central India.

BackgroundThis study was undertaken in two Primary Health Centers (PHCs) of malaria endemic district Jabalpur in Madhya Pradesh (Central India).MethodsIn this study we had investigated the relative frequencies of the different anopheline species collected within the study areas by using indoor resting catches, CDC light trap and human landing methods. Sibling species of malaria vectors were identified by cytogenetic and molecular techniques. The role of each vector and its sibling species in the transmission of the different Plasmodium species was ascertained by using sporozoite ELISA.ResultsA total of 52,857 specimens comprising of 17 anopheline species were collected by three different methods (39,964 by indoor resting collections, 1059 by human landing and 11,834 by CDC light trap). Anopheles culicifacies was most predominant species in all collections (55, 71 and 32% in indoor resting, human landing and light trap collections respectively) followed by An. subpictus and An. annularis. All five sibling species of An. culicifacies viz. species A, B, C, D and E were found while only species T and S of An. fluviatilis were collected. The overall sporozoite rate in An. culicifacies and An. fluviatilis were 0.42% (0.25% for P. falciparum and 0.17% for P. vivax) and 0.90% (0.45% for P. falciparum and 0.45% for P. vivax) respectively. An. culicifacies and An. fluviatilis were found harbouring both P. vivax variants VK-210 and VK-247, and P. falciparum. An. culicifacies sibling species C and D were incriminated as vectors during most part of the year while sibling species T of An. fluviatilis was identified as potential vector in monsoon and post monsoon season.Conclusions An. culicifacies species C (59%) was the most abundant species followed by An. culicifacies D (24%), B (8.7%), E (6.7%) and A (1.5%). Among An. fluviatilis sibling species, species T was common (99%) and only few specimens of S were found. Our study provides crucial information on the prevalence of An. culicifacies and An. fluviatilis sibling species and their potential in malaria transmission which will assist in developing strategic control measures against these vectors.

Parasites and vectors of malaria on Rusinga Island, Western Kenya

BackgroundThere is a dearth of information on malaria endemicity in the islands of Lake Victoria in western Kenya. In this study malaria prevalence and Plasmodium sporozoite rates on Rusinga Island were investigated. The contribution of different Anopheles species to indoor and outdoor transmission of malaria was also determined.MethodsActive case detection through microscopy was used to diagnose malaria in a 10% random sample of the human population on Rusinga Island and a longitudinal entomological survey conducted in Gunda village in 2012. Nocturnally active host-seeking mosquitoes were captured indoors and outdoors using odour-baited traps. Anopheles species were tested for the presence of Plasmodium parasites using an enzyme linked immunosorbent assay. All data were analyzed using generalized linear models.ResultsSingle infections of Plasmodium falciparum (88.1%), P. malariae (3.96%) and P. ovale (0.79%) as well as multiple infections (7.14%) of these parasites were found on Rusinga Island. The overall malaria prevalence was 10.9%. The risk of contracting malaria was higher among dwellers of Rusinga West than Rusinga East locations (Odds Ratio [OR] = 1.5, 95% Confidence Interval [CI] 1.14 – 1.97, P = 0.003). Parasite positivity was significantly associated with individuals who did not use malaria protective measures (OR = 2.65, 95% CI 1.76 – 3.91, p < 0.001). A total of 1,684 mosquitoes, including 74 anophelines, were captured. Unlike Culex species, more of which were collected indoors than outdoors (P < 0.001), the females of An. gambiae s.l. (P = 0.477), An. funestus s.l. (P = 0.153) and Mansonia species captured indoors versus outdoors were not different. The 46 An. gambiae s.l. collected were mainly An. arabiensis (92.3%). Of the 62 malaria mosquitoes tested, 4, including 2 indoor and 2 outdoor-collected individuals had Plasmodium.ConclusionThe rather significant and unexpected contribution of P. malariae and P. ovale to the overall malaria prevalence on Rusinga Island underscores the epidemiological importance of these species in the big push towards eliminating malaria. Although current entomological interventions mainly target indoor environments, additional strategies should be considered to prevent outdoor transmission of malaria.

Bayesian variable selection in modelling geographical heterogeneity in malaria transmission from sparse data: an application to Nouna Health and Demographic Surveillance System (HDSS) data, Burkina Faso.

BackgroundQuantification of malaria heterogeneity is very challenging, partly because of the underlying characteristics of mosquitoes and also because malaria is an environmentally driven disease. Furthermore, in order to assess the spatial and seasonal variability in malaria transmission, vector data need to be collected repeatedly over time (at fixed geographical locations). Measurements collected at locations close to each other and over time tend to be correlated because of common exposures such as environmental or climatic conditions. Non- spatial statistical methods, when applied to analyze such data, may lead to biased estimates. We developed rigorous methods for analyzing sparse and spatially correlated data. We applied Bayesian variable selection to identify the most important predictors as well as the elapsing time between climate suitability and changes in entomological indices.MethodsBayesian geostatistical zero-inflated binomial and negative binomial models including harmonic seasonal terms, temporal trends and climatic remotely sensed proxies were applied to assess spatio-temporal variation of sporozoite rate and mosquito density in the study area. Bayesian variable selection was employed to determine the most important climatic predictors and elapsing (lag) time between climatic suitability and malaria transmission. Bayesian kriging was used to predict mosquito density and sporozoite rate at unsampled locations. These estimates were converted to covariate and season-adjusted maps of entomological inoculation rates. Models were fitted using Markov chain Monte Carlo simulation.ResultsThe results show that Anophele. gambiae is the most predominant vector (79.29%) and is more rain-dependant than its sibling Anophele. funestus (20.71%). Variable selection suggests that the two species react differently to different climatic conditions. Prediction maps of entomological inoculation rate (EIR) depict a strong spatial and temporal heterogeneity in malaria transmission risk despite the relatively small geographical extend of the study area.ConclusionMalaria transmission is very heterogeneous over the study area. The EIR maps clearly depict a strong spatial and temporal heterogeneity despite the relatively small geographical extend of the study area. Model based estimates of transmission can be used to identify high transmission areas in order to prioritise interventions and support research in malaria epidemiology.Electronic supplementary materialThe online version of this article (doi:10.1186/s13071-015-0679-7) contains supplementary material, which is available to authorized users.

Malaria incidence and assessment of entomological indices among resettled communities in Ethiopia: a longitudinal study.

BackgroundPopulation resettlement has been considered among factors that may increase risk of malaria transmission. This study reports, the impact of resettlement on malaria incidence and entomological indices among communities resettled in suburbs of Jimma town, southwestern Ethiopia.MethodsA cohort of 604 study participants (302 resettlers and 302 non-resettlers) was monthly followed-up from September to November 2013 using active case detection. Moreover, longitudinal entomological study was conducted from June to November 2013. Anopheline mosquitoes were collected using CDC light traps and pyrethrum spray catches. Sporozoite ELISA was performed to determine Plasmodium infection rates.ResultsOverall, 112 malaria cases were recorded during the three-month follow-up, of which 74.1% of the cases were from resettlement villages. Plasmodium falciparum incidence from resettlement and non-resettlement villages was 52.5 and 14.5/1,000 person-months at risk, respectively. Resettlement villages were three times at higher risk of Plasmodium infection (OR = 2.8, 95% CI: 1.22-6.48). Anopheles gambiae s.l. was the predominant (86.6%) of all the collected anopheline mosquito species. Plasmodium sporozoite rate in the resettlement and non-resettlement villages was 2.1 and 0.72%, respectively. Plasmodium falciparum entomological inoculation rate (EIR) for An. gambiae s.l. in the resettlement and non-resettlement villages was 13.1 and 0 infective bites/person/night, respectively. Both sporozoite rate and EIR were significantly higher in the resettlement villages (p < 0.05).ConclusionResettled communities were at higher risk of malaria infection as compared to non-resettled communities. Special attention should be given to malaria control interventions during resettlement programmes.