Human Blood Index Research Articles

Estimation of the Human Blood Index in Malaria Mosquito Vectors in Equatorial Guinea after Indoor Antivector Interventions

We determined the Human Blood Index (HBI) of malaria mosquito vectors in Equatorial Guinea. We used a polymerase chain reaction (PCR)-based methodology to identify blood meal sources in engorged mosquitoes. We observed high HBI values, indicating that these vectors are highly anthropophilic despite intensive intradomicillary application of residual insecticides. Our results suggest that estimating the HBI can be a relatively simple and easy way to evaluate the efficacy of antimalaria interventions where an observed diversion to non-human hosts may successfully contribute to the interruption of malaria transmission.

Daily survival and human blood index of major malaria vectors associated with oil palm cultivation in Cameroon and their role in malaria transmission

To describe the daily survival rate, biting habits, feeding behaviour, sporozoite and entomological inoculation rates of Anopheles species and their relative contribution to Plasmodium falciparum transmission in large-scale oil palm-growing zones in Cameroon. Entomological surveys by employing human-landing catches, both indoors and outdoors, two nights per month for 12 months from October 2004. A total of 2485 Anopheles were collected and four species recorded: An. funestus, An. hancocki, An. gambiae s.s. and An. nili. There was substantial indoors to outdoors variation and seasonality in the distribution of the Anopheles population. All four species showed similar nocturnal biting patterns and were sympatrically involved in malaria transmission simultaneously or replacing each other seasonally. Their constant presence throughout the year and longevity suggest that transmission can occur at any time if parasite reservoirs are present in the area. The circumsporozoite protein rates were 5.8%, 6.3%, 8.4% and 4.1%; mean anthropophilic rates were 97.1%, 94.5%, 73.9% and 77.6%, respectively, for An. funestus, An. gambiae s.l, An. nili and An. hancocki. The annual entomological inoculation rate was 1077 infectious bites per person. Of those An. gambiae specimens testing positive for P. falciparum sporozoites, 69.01% were M form and 30.99% were S form. Any vector control strategy intending to significantly reduce the malaria burden in the palm estate will have to take into account entomological heterogeneity in addition to ecological and socio-economic multiplicity.

Studies on seasonal fluctuation of different indices related to filarial vector, Culex quinquefasciatus around foothills of Susunia of West Bengal, India

Objective To collect information on the seasonal activity of filarial vector around foothills of Susunia of West Bengal, India. Methods Indoor-resting mosquitoes were collected from the human habitations of 16 villages. Collected mosquitoes were identified and examined for different parameters following standard methods recommended by World Health Organization and pioneer workers. Results Overall man hour density, infection rate, infectivity rate, human blood index and daily mortality rate of the vector were assessed as 10.54%, 6.31%, 1.38%, 77.33% and 13.00%, respectively, which were 12.17%, 8.63%, 2.26%, 100.00% and 17.00%, respectively in the summer season. Average load of microfilaria, 1 st stage, 2 nd stage and 3 rd stage larvae of Wuchereria bancrofti in infected vectors were 10.02, 7.50, 5.56 and 4.68, respectively, which were 11.58, 9.90, 6.27 and 5.44, respectively in summer. Among the searched shelters 63.37%, 8.16% and 2.08% were found to be invaded by any vector, infected vector and infective vector, respectively, which were 68.75%, 12.76% and 3.91%, respectively in summer. Conclusions Different indices related to the vector were much higher in summer, which indicates that, summer is the most favorable season for transmission of lymphatic filariasis in the study area. Available data will help to formulate an effective vector control measure.

Wash-resistance and field evaluation of alphacypermethrin treated long-lasting insecticidal net (Interceptor ®) against malaria vectors Anopheles culicifacies and Anopheles fluviatilis in a tribal area of Orissa, India

A field trial was conducted on the efficacy of Interceptor nets—a long-lasting insecticidal net (LLN) factory treated with alphacypermethrin 0.667% (w/w) corresponding to 200 mg/m 2, against malaria vectors Anopheles culicifacies and Anopheles fluviatilis in one of the highly endemic areas of Orissa. The study area comprised 19 villages which were randomized into three clusters and designated as Interceptor net cluster, untreated net cluster, and no net cluster. Baseline studies showed that both the vector species An. culicifacies and An. fluviatilis were 100% susceptible to alphacypermethrin. Results of wash-resistance and bio-efficacy of Interceptor nets showed 100% mortality in An. culicifacies and An. fluviatilis even after 20 washings. Bioassays on the Interceptor nets while in use in the field conditions showed a knockdown effect on 70–90% mosquitoes during different months of intervention after 3 min of exposure and 100% mortality was recorded after 24 h of recovery period. The median knockdown time for these species ranged between 4.10–5.25 min and 4.00–5.00 min respectively during intervention period. In Interceptor net study area, there was a significant reduction of 88.9, 96.3 and 90.6% in the entry rate of An. culicifacies, An. fluviatilis and other anopheline species respectively with an over all reduction of 87.5% in total mosquitoes. The overall feeding success rate of mosquitoes in the trial villages was only 12.8% in comparison to 35.0 and 78.8% in villages with untreated nets and no nets respectively. A significant reduction was also recorded in parity rate and human blood index of vector species in Interceptor net area. The results of the study showed that Interceptor nets are effective against the malaria vectors and may be used as a suitable intervention strategy in high-risk areas.

Host-Feeding Patterns ofAedes albopictus(Diptera: Culicidae) in Urban and Rural Contexts within Rome Province, Italy

Knowledge of the frequency of contact between a mosquito species and its different hosts is essential to understand the role of each vector species in the transmission of diseases to humans and/or animals. However, no data are so far available on the feeding habits of Aedes albopictus in Italy or in other recently colonized temperate regions of Europe, due to difficulties in collecting blood-fed females of this diurnal and exophilic species. We analyzed Ae. albopictus host-feeding patterns in two urban and two rural sites within the area of Rome (Italy). Ae. albopictus was collected using sticky-traps and the blood-meal origin of 303 females was determined by direct dot-ELISA. The blood-fed sample, although representing only 4% of the total Ae. albopictus collected, demonstrates the useful application of sticky-trap in studying the feeding behavior of the species. The human blood index was significantly different among sites, ranging from 79-96% in urban sites to 23-55% in rural sites, where horses and bovines represented the most bitten hosts. The results obtained confirm the plastic feeding behavior shown by Ae. albopictus in its original range of distribution and highlights the high potential of this species as a vector of human pathogens in urban areas of Italy, where both humans and the mosquito itself may reach very high densities.

Host-Feeding Preference of the Mosquito,Culex quinquefasciatus, in Yucatan State, Mexico

Studies were conducted to determine the host-feeding preference of Culex quinquefasciatus Say (Diptera: Culicidae) in relation to the availability of human and domestic animals in the city of Merida, Yucatan State, Mexico. Mosquitoes were collected in the backyards of houses using resting wooden boxes. Collections were made five times per week from January to December 2005. DNA was extracted from engorged females and tested by PCR using universal avian- and mammalian-specific primers. DNA extracted from avian-derived blood was further analyzed by PCR using primers that differentiate among the birds of three avian orders: Passeriformes, Columbiformes and Galliformes. PCR products obtained from mammalian-derived blood were subjected to restriction enzyme digestion to differentiate between human-, dog-, cat-, pig-, and horse-derived blood meals. Overall, 82% of engorged mosquitoes had fed on birds, and 18% had fed on mammals. The most frequent vertebrate hosts were Galliformes (47.1%), Passeriformes (23.8%), Columbiformes (11.2%) birds, and dogs (8.8%). The overall human blood index was 6.7%. The overall forage ratio for humans was 0.1, indicating that humans were not a preferred host for Cx. quinquefasciatus in Merida.

Climate change and altitudinal structuring of malaria vectors in south-western Cameroon: their relation to malaria transmission

An entomological survey was conducted in Cameroon between October 2004 and September 2005, in nine localities targeted for malaria vector control based on adult productivity and variability. Mosquitoes were collected by human-landing catches (HLCs) and pyrethrum spray catches. A total of 12 500 anophelines were collected and dissected: Anopheles gambiae s.l. (56.86%) , An. funestus s.l. (32.57%) , An. hancocki (9.38%) , and An. nili (1.18%). Applying PCR revealed that specimens of the An. funestus group were An. funestus s.s. and An. gambiae complex were mostly An. melas and An. gambiae s.s. of the M and S molecular forms with the M forms being the most predominant. The natural distribution patterns of Anopheles species were largely determined by altitude with some species having unique environmental tolerance limits. A human blood index (HBI) of 99.05% was recorded. Mean probability of daily survival of the malaria vectors was 0.92, with annual mean life expectancy of 21.9 days and the expectation of infective life was long with a mean of 7.4 days. The high survival rates suggest a high vector potential for the species. This information enhances the development of a more focused and informed vector control intervention.

Seasonal Abundance and Host-Feeding Patterns of Anopheline Vectors in Malaria Endemic Area of Iran

Seasonal abundance and tendency to feed on humans are important parameters to measure for effective control of malaria vectors. The objective of this study was to describe relation between feeding pattern, abundance, and resting behavior of four malaria vectors in southern Iran. This study was conducted in ten indicator villages (based on malaria incidence and entomological indices) in mountainous/hilly and plain regions situated south and southeastern Iran. Mosquito vectors were collected from indoor as well as outdoor shelters and the blood meals were examined by ELISA test. Over all 7654 female Anopheles spp. were captured, the most common species were Anopheles stephensi, An. culicifacies, An. fluviatilis, and An. d'thali. The overall human blood index was 37.50%, 19.83%, 16.4%, and 30.1% for An. fluviatilis, An. stephensi, An. culicifacies, and An. d'thali, respectively. In addition, An. fluviatilis fed on human blood during the entire year but the feeding behavior of An. stephensi and An. culicifacies varied according to seasons. Overall, the abundance of the female mosquito positive to human blood was 4.25% per human shelter versus 17.5% per animal shelter. This result indicates that the vectors had tendency to rest in animal shelters after feeding on human. Therefore, vector control measure should be planned based on such as feeding pattern, abundance, and resting behavior of these vectors in the area.

Effect of untreated bed nets on blood-fed Phlebotomus argentipes in kala-azar endemic foci in Nepal and India

Observational studies in the Indian subcontinent have shown that untreated nets may be protective against visceral leishmaniasis (VL). In this study, we evaluated the effect of untreated nets on the blood feeding rates of Phlebotomus argentipes as well as the human blood index (HBI) in VL endemic villages in India and Nepal. The study had a 'before and after intervention' design in 58 households in six clusters. The use of untreated nets reduced the blood feeding rate by 85% (95% CI 76.5-91.1%) and the HBI by 42.2% (95% CI 11.1-62.5%). These results provide circumstantial evidence that untreated nets may provide some degree of personal protection against sand fly bites.

Spatial distribution, blood feeding pattern, and role of Anopheles funestus complex in malaria transmission in central Kenya

Studies were conducted to determine the role of sibling species of Anopheles funestus complex in malaria transmission in three agro-ecosystems in central Kenya. Mosquitoes were sampled indoors and outdoors, and rDNA PCR was successfully used to identify 340 specimens. Anopheles parensis (91.8%), A. funestus (6.8%), and Anopheles leesoni (1.5%) were the three sibling species identified. A. parensis was the dominant species at all study sites, while 22 of 23 A. funestus were collected in the non-irrigated study site. None of the 362 specimens tested was positive for Plasmodium falciparum circumsporozoite proteins by enzyme-linked immunosorbent assay. The most common blood-meal sources (mixed blood meals included) for A. parensis were goat (54.0%), human (47.6%), and bovine (39.7%), while the few A. funestus s.s. samples had fed mostly on humans. The human blood index (HBI) for A. parensis (mixed blood meals included) in the non-irrigated agro-ecosystem was 0.93 and significantly higher than 0.33 in planned rice agro-ecosystem. The few samples of A. funestus s.s. and A. funestus s.l. also showed a trend of higher HBI in the non-irrigated agro-ecosystem. We conclude that agricultural practices have significant influence on distribution and blood feeding behavior of A. funestus complex. Although none of the species was implicated with malaria transmission, these results may partly explain why non-irrigated agro-ecosystems are associated with higher risk of malaria transmission by this species compared to irrigated agro-ecosystems.

Field Evaluation of Olyset Nets: A Long-Lasting Insecticidal Net Against Malaria VectorsAnopheles culicifaciesandAnopheles fluviatilisin a Hyperendemic Tribal Area of Orissa, India

A village-scale trial was conducted on the efficacy of Olyset nets: a long-lasting insecticidal net (LLIN) factory treated with 2% wt:wt permethrin against malaria vectors Anopheles culicifacies Giles and Anopheles fluviatilis James, in Sundargarh District, Orissa, India. The study area comprised 22 villages that were randomized into three clusters and designated as Olyset net, untreated net, and no net clusters. Baseline studies showed that both vector species were 100% susceptible to permethrin. Results of wash resistance and bioefficacy of Olyset nets showed 100% mortality in An. culicifacies up to 11 washings, whereas 100% mortality was observed in An. fluviatilis even after 20 washings. The median knock-down time for these species ranged between 4.55-6.00 and 4.45-5.45 min, respectively, during 1 yr of intervention. In the Olyset net study area, there was a significant reduction of 80.6, 94.1, and 76.7% in the entry rate of An. culicifacies, An. fluviatilis, and other anopheline species, respectively, with an overall reduction of 63.5% in total mosquitoes. Floor sheet collections in houses with Olyset nets indicated 39% immediate mortality in total mosquitoes. The overall feeding success rate of mosquitoes in the trial village was only 18.0% in comparison to 44.2 and 79.1% in villages with untreated nets and no nets, respectively. A significant reduction was also recorded in parity rate and human blood index of vector species in the Olyset net area. This study showed that Olyset nets are an effective personal protection tool that can be used in a community-based intervention program.

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.

Host choice and multiple blood feeding behaviour of malaria vectors and other anophelines in Mwea rice scheme, Kenya

BackgroundStudies were conducted between April 2004 and February 2006 to determine the blood-feeding pattern of Anopheles mosquitoes in Mwea Kenya.MethodsSamples were collected indoors by pyrethrum spay catch and outdoors by Centers for Disease Control light traps and processed for blood meal analysis by an Enzyme-linked Immunosorbent Assay.ResultsA total of 3,333 blood-fed Anopheles mosquitoes representing four Anopheles species were collected and 2,796 of the samples were assayed, with Anopheles arabiensis comprising 76.2% (n = 2,542) followed in decreasing order by Anopheles coustani 8.9% (n = 297), Anopheles pharoensis 8.2% (n = 272) and Anopheles funestus 6.7% (n = 222). All mosquito species had a high preference for bovine (range 56.3–71.4%) over human (range 1.1–23.9%) or goat (0.1–2.2%) blood meals. Some individuals from all the four species were found to contain mixed blood meals. The bovine blood index (BBI) for An. arabiensis was significantly higher for populations collected indoors (71.8%), than populations collected outdoors (41.3%), but the human blood index (HBI) did not differ significantly between the two populations. In contrast, BBI for indoor collected An. funestus (51.4%) was significantly lower than for outdoor collected populations (78.0%) and the HBI was significantly higher indoors (28.7%) than outdoors (2.4%). Anthropophily of An. funestus was lowest within the rice scheme, moderate in unplanned rice agro-ecosystem, and highest within the non-irrigated agro-ecosystem. Anthropophily of An. arabiensis was significantly higher in the non-irrigated agro-ecosystem than in the other agro-ecosystems.ConclusionThese findings suggest that rice cultivation has an effect on host choice by Anopheles mosquitoes. The study further indicate that zooprophylaxis may be a potential strategy for malaria control, but there is need to assess how domestic animals may influence arboviruses epidemiology before adapting the strategy.

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.

Culex nigripalpus Theobald (Diptera, Culicidae) feeding habit at the Parque Ecológico do Tietê, São Paulo, Brazil

The blood feeding of a population of Cx. nigripalpus from Parque Ecológico do Tietê (PET) was investigated using an indirect ELISA protocol. Mosquitoes were captured outside houses. Five hundred sixteen engorged females collected in a reforested area and 25 in an open area were tested. Rodents and dogs were the most common blood sources, accounting for approximately 65.3% of blood meals. Human blood was detected in 10.9%, dog blood in 26.1%, chicken blood in 2.4%, and rodent blood in 39.2% of the 541 insects tested. ELISA failed in identifying the blood sources of 233 engorged females, indicating that the mosquitoes may have fed on a host which was not tested. One hundred six individuals were positive for more than one host. The unweighted human blood index was 0.14 and the rodent/human, human/chicken, and dog/rodent feeding index values were 2.70, 1.51, and 1.33, respectively. Furthermore, rodents are defensive hosts for this haematophagous insect which looks for another host to complete blood-feeding. Considering that rodents are potential reservoirs for Mucambo virus and Saint Louis encephalitis virus and that Cx. nigripalpus feed on the blood of those mammals, we hypothesize that mosquito population in PET could participate in the transmission cycle of those arboviruses. Additionally, this species might be involved in the transmission of Dirofilaria immitis to dogs at this area.

Lymphatic filariasis in the coastal areas of Digha, West Bengal, India

The state of West Bengal, India, has a long coastline with the Bay of Bengal. No information exists regarding filarial epidemiology and its vector in these coastal areas. The present study was designed to assess the epidemiology of lymphatic filariasis and the role of available mosquitoes as its vector in eight coastal villages around Digha, West Bengal. Night blood samples of 4016 individuals were collected and each of them was examined clinically for any manifestations of the disease. Overall, microfilaria rate, mean microfilarial density and disease rate were 9.06%, 8.63% and 7.72%, respectively. The causative parasite was identified as Wuchereria bancrofti and Culex quinquefasciatus was incriminated as the vector responsible. Vector infection and infectivity rates were assessed to be 12.5% and 0.73%, respectively. The human blood index of human-house-frequenting vector population was 70%. Vector density, vector infection, infectivity rates and human blood index were higher in the rainy season in the study area. Overall, the filarial situation was bad and, as a measure, single-dose diethylcarbamazine citrate (6 mg/kg body weight) treatment was given to all the microfilariaemic patients. Night blood samples of the treated individuals were tested for microfilariae on days 10 and 365, which revealed interesting results.

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

SPATIAL ANALYSIS OF MALARIA TRANSMISSION PARAMETERS IN THE RICE CULTIVATION AREA OF OFFICE DU NIGER, MALI

The effects of rice growth environment on malaria transmission, taking into account spatial correlation, were assessed in the Office du Niger, Mali. Between April 1999 to January 2001, 8 quarterly entomologic surveys were conducted in 18 villages in 3 agricultural zones. Vector densities in sleeping houses were related to rice crop, rice development stages, vegetation abundance, water state, and seasons. They were high throughout the rice-growing seasons, increased as the rice crop developed, and decreased as vegetation became abundant. They also showed large spatial correlations (up to 30.6 km). The vectorial capacity exhibited both seasonal and village-to-village variation. Parity and the human blood index were weakly related to adult densities and showed low spatial correlations (up to 3.4 km), which suggested that small area variation in malaria transmission results mainly from variations in vector-human contact. Control strategies in rice cultivation areas should pay attention to this local variation.

Identification of four members of the Anopheles funestus (Diptera: Culicidae) group and their role in Plasmodium falciparum transmission in Bagamoyo coastal Tanzania

The role of Anopheles funestus group in malaria transmission was investigated in Bagamoyo coastal Tanzania, in the process of characterizing the area as a malaria vaccine testing site. Mosquitoes were sampled inside houses and multiplex PCR was used to identify 649 specimens. The following species were found: A. funestus s.s. (84.3%), A. leesoni (13.6%), A. rivulorum (1.5%) and A. parensis (0.6%). Multiplex PCR of 147 blood-fed specimens showed that over half (57.1%) of the identifiable blood meals were taken from human hosts, and human blood index in A. funestus and A. leesoni was 55% and 82% respectively. Plasmodium falciparum infection rate determined by nested PCR was 11% in A. funestus s.s. Although the abundance was low, 26 specimens of A. leesoni, two of A. rivolurum and one of A. parensis were found positive for P. falciparum. The presence of four A. funestus species in Tanzania emphasizes the relevance to define precisely their spatial and temporal distribution, specific behaviour, ecology and exact role in malaria transmission.