Pit Shelters Research Articles

Methods of sampling malaria vectors and their reliability in estimating entomological indices in Africa.

In efforts to intensify malaria control through vector control and hasten the progress towards elimination, the impact of control interventions needs to be evaluated. This requires sampling vector population using appropriate trapping methods. The aim of this article is to critically review methods of sampling malaria vectors and their reliability in estimating entomological indicators of malaria transmission in Africa. The standard methods are human landing catch (HLC), pyrethrum spray catch, and pit shelter for sampling host-seeking, indoor resting, and outdoor resting malaria vectors, respectively. However, these methods also have drawbacks such as exposure of collectors to infective mosquito bites, sampling bias, and feasibility issue. Centers for Disease Control and Prevention (CDC) light traps placed beside human-occupied bed nets have been used as an alternative to the HLC for sampling host-seeking malaria vectors. Efforts have been made to evaluate the CDC light traps against HLC to generate a conversion factor in order to use them as a proxy estimator of human biting rate and entomological inoculation rates in Africa. However, a reproducible conversion factor was not found, indicating that the trapping efficiency of the CDC light traps varies between different geographical locations. Several other alternative traps have also been developed and evaluated in different settings but most of them require further standardization. Among these, human-baited double net trap/CDC light trap combination and mosquito electrocuting trap have the potential to replace the HLC for routine malaria vector surveillance. Further research is needed to optimize the alternative sampling methods and/or develop new surveillance tools based on vector behavior.

Cattle feeding tendency of Anopheles mosquitoes and their infection rates in Aradum village, North Wollo, Ethiopia: an implication for animal-based malaria control strategies

BackgroundSurveillance of indoor and outdoor resting malaria vector populations is crucial to monitor possible changes in vector resting and feeding behaviours. This study was conducted to assess the resting behaviour, blood meal sources and circumsporozoite (CSP) rates of Anopheles mosquito in Aradum village, Northern Ethiopia.MethodsMosquito collection was conducted from September 2019 to February 2020 using clay pots (indoor and outdoor), pit shelter and pyrethrum spray catches (PSC). The species of Anopheles gambiae complex and Anopheles funestus group were identified using polymerase chain reaction (PCR). Enzyme-linked immunosorbent assay (ELISA) was done to determine CSP and blood meal sources of malaria vectors.ResultsA total of 775 female Anopheles mosquitoes were collected using the clay pot, PSC and pit shelter. Seven Anopheles mosquito species were identified morphologically, of which Anopheles demeilloni (593; 76.5%) was the dominant species followed by An. funestus group (73; 9.4%). Seventy-three An. funestus group screened by PCR, 91.8% (67/73) were identified as Anopheles leesoni and only 2.7% (2/73) were found to be Anopheles parensis. The molecular speciation of 71 An. gambiae complex confirmed 91.5% (65/71) of Anopheles arabiensis. The majority of Anopheles mosquitoes were collected from outdoor pit shelter (42.2%) followed by outdoor clay pots. The majority of the blood meal of An. demeilloni (57.5%; 161/280), An. funestus sensu lato 10 (43.5%) and An. gambiae (33.3%; 14/42) originated from bovine. None of the 364 Anopheles mosquitoes tested for Plasmodium falciparum and Plasmodium vivax sporozoite infections were positive.ConclusionSince the Anopheles mosquitoes in the area prefer to bite cattle, it may be best to target them with an animal-based intervention. Clay pots could be an alternative tool for outdoor monitoring of malaria vectors in areas where pit shelter construction is not possible.

Outdoor and early hour human biting activities of malaria mosquitoes and the suitability of clay pot for outdoor resting mosquito collection in malaria endemic villages of southern Rift Valley, Ethiopia

BackgroundSampling adult Anopheles mosquitoes is important for assessing vector density, estimating the sporozoite infection rate, and quantifying the impact of vector control interventions. The objective of this study was to assess the Anopheles mosquito species composition, and their outdoor and indoor biting activities, and to evaluate the suitability of clay pots for indoor and outdoor resting mosquito collections. MethodsTwo malaria-endemic villages in the Gamo zone were purposely selected. Forty clay pots were deployed for outdoor resting mosquitoes sampling and another forty for indoor resting sampling. Twenty pit shelters were constructed for outdoor resting mosquito collection. The human landing catch (HLC) technique was employed to collect indoor and outdoor host-seeking mosquitoes in two households in each village. Morphological identification of the Anopheles mosquito was done using an identification key. Enzyme-linked immunosorbent assay technique was used for blood meal origin and circumsporozoite proteins (CSP) test. Speciation of An. gambiae complex was done using polymerase chain reaction. A Chi-square test was used to compare the effectiveness of clay pot and pit shelters for outdoor resting sampling. ResultsA total of 904 female Anopheles mosquitoes comprising An. gambiae complex, An. pharoensis, An. tenebrosus, An. dencalicus and An. demelloni were sampled. The majority (64%) of them were sampled by the HLC technique. There was a slight difference between the outdoor clay pot (19%) and pit shelter (17%) collection. No Anopheles mosquitoes were collected indoor using clay pots. All mosquitoes were tested for CSPs, but none of them were found to be positive. Anopheles mosquitoes were tending to bite humans outdoor than indoors, and their peak biting hours was 10–11 pm. The human blood meal index of Anopheles mosquitoes was 0.07 from pit shelters and it was 0.04 from clay pots. The bovine blood meal index was 0.45 for mosquitoes from both pit shelters and clay pot collections. ConclusionAnopheles arabiensis was the predominant species and it was tending to bite cattle more than humans. Clay pot could be suitable for outdoor resting mosquito collection, but not for indoor resting species.

Investigation on Mosquitoes Fauna (Diptera: Culicidae) and Probable Vector of West Nile Virus in Lorestan Province, Western Iran.

Fauna and larval habitat characteristics studies on mosquitoes are important tools to identify the breeding places of the vectors and management of the control strategies. This study was done to provide data on Culicidae fauna, larval habitat characteristics and identifying potential vectors of West Nile virus in Lorestan Province, west of Iran. Culicidae mosquitoes were collected at three counties and nine site stations from Lorestan Province, west of Iran in 2017. Adult mosquitoes were collected using human and animal bite collection methods, New Jersey and CDC light traps and pit shelters by aspirator. Larva were collected by dipping method. RT-PCR technique was employed for detection of the West Nile virus among mosquito's samples. 4805 mosquitoes were collected from three counties and nine sites in Lorestan Province during June-October 2017, including 4363 adults and 442 larvae. The most abundant species collected from all counties in both adult and larval stages were Culex pipiens (49.10%), Cx. theileri (31.82%), Anopheles maculipennis (11.09%), An. superpictus (2.66%), An. stephensi (2.12%), Cx. perexiguus (1.89%), An. dthali (1.17%) and An. sacharovi (0.15%) respectively. West Nile virus was detected in none of mosquitoes examined. The results of this study revealed that arbovirus vectors such as Cx. pipiens along with Cx. theileri and Cx. perexiguus are well adapted to a broad range of habitats and different climatic conditions in Lorestan Province. That necessitates further routine surveillance of arboviral infections.

Population Fluctuations and Abundance Indices of Mosquitoes (Diptera: Culicid), as the Potential Bridge Vectors of Pathogens to Humans and Animals in Mazandaran Province, Northern Iran.

Background:Seasonal activity patterns of mosquitoes are essential as baseline knowledge to understand the transmission dynamics of vector-borne diseases. This study was conducted to evaluate the monthly dynamics of the mosquito populations and their relation to meteorological factors in Mazandaran Province, north of Iran.Methods:Mosquito adults and larvae were collected from 16 counties of Mazandaran Province using different sampling techniques, once a month from May to December 2014. “Index of Species Abundance” (ISA) along with “Standardized ISA” (SISA) was used for assessing the most abundant species of mosquitoes based on the explanations of Robert and Hsi. Pearson’s correlation coefficient (R) was used to assess the relationships between the monthly population fluctuations and meteorological variables.Results:Overall, 23750 mosquitoes belonging to four genera and nineteen species were collected and identified. The highest population density of mosquitoes was in July and the lowest in May. The ISA/SISA indices for Culex pipiens were both 1 for larvae and 1.25/0.973 for adults in total catch performed in human dwellings. For Cx. tritaeniorhynchus, the ISA/SISA were 1.68/0.938 in pit shelter method. A significant positive correlation was observed between population fluctuations of Cx. tritaeniorhynchus and mean temperature (R: 0.766, P< 0.027).Conclusion:The results indicated that the mosquitoes are more active in July, and Cx. pipiens and Cx. tritaeniorhynchus were the most abundant species. Considering the potential of these species as vectors of numerous pathogens, control programs can be planed based on their monthly activity pattern in the area.

Assessment of Malaria Vector Behavior in Selected Sentinel Sites of Ethiopia

In Ethiopia, malaria incidence has significantly reduced in the past decade through the combined use of conventional vector control approaches and antimalarial drugs. However, the sustainability of this achievement is threatened by the shift in biting and resting behaviors and emergence of insecticide resistance by the primary malaria vector. Therefore, continuous monitoring of the behaviour of malaria mosquitoes in different sentinel sites is crucial to design effective prevention and control methods in the local context. Entomological investigations were conducted in three sentinel sites for five consecutive months during the major malaria transmission season. The species composition, population dynamics, biting and resting behaviours of malaria vectors were determined using center for disease control and prevention (CDC) light trap, human landing catch (HLC), pyrethrum spray catch (PSC) and Pit shelter collection (PFS). Accordingly, 10 households for CDC, 10 households for PSC, 10 households for PFS and 5 households for HLC from each site were randomly enrolled for mosquito collection. A total of 8,297 anopheline mosquitoes were collected from the three sites, out of which 4,525 (54.5%) were An. gambiae, s.l. 2,028 (24.4%) were An. pharoensis, 160 (1.9%) were An. funestus and the rest 1,584 (19%) were other Anophelines (An. coustani, An. cinerus and An. tenebrosus). No significant variation (P = 0.476) was observed between indoor (25.2/trap-night and outdoor collections (20.1/trap-night). Six hundred seventy six (43.3%) of An. gamiabiae s.l. (primary vector) were collected between 18:00 and 22:00 hours. Biting activity declined between 00:00 and 02:00 hours. The national malaria control program should pay close attention to the shifting behavior of vector mosquitoes as the observed outdoor feeding tendency of the vector population could pose challenges to the indoor intervention tools IRS and LLINs. Funding Information: This work was supported by the Ministry of health (Global fund). Declaration of Interests: The authors declare that they have no conflicts of interest. Ethics Approval Statement: This study was issued ethical clearance by the Scientific and Ethical Review Committee (SERC) in Ethiopia Public Health Institute with reference number EPHI 6.13/976. Verbal consent was obtained from the heads of households before commencing mosquito collection in houses. The heads of households surveyed were informed that participation in the study was absolutely voluntary and that even after consenting they would be free to withdraw from the study at any time.

Species diversity and distribution of Anopheles mosquitoes in Bure district, Northwestern Ethiopia

Malaria is one the leading health problem of the Ethiopia. Previously, areas above 2,000 m elevation were considered as malaria free areas. However, the major malaria epidemics were seen in areas at an altitude up to 3,000 m above sea level. These epidemics were due to climate and land-use changes (ecological changes) and still malaria is a growing health problem in highland parts of Ethiopia. This study aimed to investigate the species diversity, abundance and distribution of Anopheles mosquitoes in highland fringe of Bure district, Northwestern Ethiopia. It was done in the three different agroecological villages, Bukta (Irrigated), Workimdr (non-irrigated with few dry season breeding habitats) and Shnebekuma (non-irrigated with many dry season breeding habitats). Anopheles mosquitoes were collected by the Centers for Disease Control and Prevention Light Traps Catches, Pyrethrum Spray Catches, and Artificial Pit Shelters (APSs) from twenty-seven houses, thirty houses, and six APSs, respectively. Anopheles mosquitoes were identified morphologically to species using standard keys. Furthermore, molecular identification of Anopheles gambiae s.l was carried out using species-specific Polymerase Chain Reaction. Independent T-Test and One-way- ANOVA were employed to compare the mean mosquito's density between villages and species, indoor and outdoor host seeking mosquitoes. Descriptive statistic was used to calculate the proportion of each Anopheles species. Nine Anopheles mosquito species were identified in the study area which includes: Anopheles demeilloni, An. arabiensis, An. funestus group, An. coustani, An. squamosus, An. cinereus, An. pharoensis, An. rupicolus, and An. natalensis. Of the 4,703 Anopheles mosquitoes collected, An. demeilloni was the most prominent (50.7%, n = 2383) whereas An. rupicolus (0.03%, n = 3), and An. natalensis (0.02%, n = 1) were the least abundant. Higher mean density of Anopheles mosquitoes was collected from the non-irrigated village (2.395 ± 0.100) than irrigated (1.351 ± 0.109) (p = 0.001). In conclusion, three of the most important malaria vectors (An. arabiensis, An. funestus group and An. pharoensis) of Ethiopia were recorded in the study sites, especially the first two was found thought-out the year. Most of the Anopheles mosquitoes were collected from non-irrigated villages. Thus, breeding habitat management must be practiced throughout the year together with long-lasting insecticide-treated nets and insecticide residual sprays.

Morphological and molecular identification of Culicidae mosquitoes (Diptera: Culicidae) in Lorestan province, Western Iran

Culicidae mosquitoes are main vectors of arboviruses that cause arboviral diseases in humans. Studies on fauna, ecology, biology, resting behaviors of Culicidae mosquitoes are important and greatly impacts the control of arboviral diseases that are transmitted by vectors. The aim of the present study was to determine fauna of mosquitoes (Diptera: Culicidae) based on morphological and molecular (genomic) identification and their habitats in Lorestan province, Western Iran. Meanwhile mosquito samples were examined for arbovirus infection.Culicidae mosquitoes were caught in 2015 and 2016 from human homes, animal dwellings, storehouses and pit shelters in Lorestan province, Western Iran, using an oral aspirator (hand catch), total catch, human and animal bait and light trap methods. The samples were identified on the genus and species. Six species of Culex and eight species of Anopheles were caught. One complex species (Cx. pipiens complex) and a hybrid between Cx. pipiens pipiens biotype pipiens and Cx. pipiens pipiens biotype molestus were identified. Among all of the trapped mosquitoes (4211), 94.68% were from genus Culex mosquitoes (3987), which indicate that this genus is the dominant in Lorestan province, Western Iran. Anopheles comprised of 201 individuals out of the total catch. Arboviruses were not detected in these samples.

Resting behaviour of malaria vectors in highland and lowland sites of western Kenya: Implication on malaria vector control measures.

Understanding the interactions between increased insecticide resistance and resting behaviour patterns of malaria mosquitoes is important for planning of adequate vector control. This study was designed to investigate the resting behavior, host preference and rates of Plasmodium falciparum infection in relation to insecticide resistance of malaria vectors in different ecologies of western Kenya. Anopheles mosquito collections were carried out during the dry and rainy seasons in Kisian (lowland site) and Bungoma (highland site), both in western Kenya using pyrethrum spray catches (PSC), mechanical aspiration (Prokopack) for indoor collections, clay pots, pit shelter and Prokopack for outdoor collections. WHO tube bioassay was used to determine levels of phenotypic resistance of indoor and outdoor collected mosquitoes to deltamethrin. PCR-based molecular diagnostics were used for mosquito speciation, genotype for knockdown resistance mutations (1014S and 1014F) and to determine specific host blood meal origins. Enzyme-linked Immunosorbent Assay (ELISA) was used to determine mosquito sporozoite infections. Anopheles gambiae s.l. was the most predominant species (75%, n = 2706) followed by An. funestus s.l. (25%, n = 860). An. gambiae s.s hereafter (An. gambiae) accounted for 91% (95% CI: 89-93) and An. arabiensis 8% (95% CI: 6-9) in Bungoma, while in Kisian, An. arabiensis composition was 60% (95% CI: 55-66) and An. gambiae 39% (95% CI: 34-44). The resting densities of An. gambiae s.l and An. funestus were higher indoors than outdoor in both sites (An. gambiae s.l; F1, 655 = 41.928, p < 0.0001, An. funestus; F1, 655 = 36.555, p < 0.0001). The mortality rate for indoor and outdoor resting An. gambiae s.l F1 progeny was 37% (95% CI: 34-39) vs 67% (95% CI: 62-69) respectively in Bungoma. In Kisian, the mortality rate was 67% (95% CI: 61-73) vs 76% (95% CI: 71-80) respectively. The mortality rate for F1 progeny of An. funestus resting indoors in Bungoma was 32% (95% CI: 28-35). The 1014S mutation was only detected in indoor resitng An. arabiensis. Similarly, the 1014F mutation was present only in indoor resting An. gambiae. The sporozoite rates were highest in An. funestus followed by An. gambiae, and An. arabiensis resting indoors at 11% (34/311), 8% (47/618) and 4% (1/27) respectively in Bungoma. Overall, in Bungoma, the sporozoite rate for indoor resting mosquitoes was 9% (82/956) and 4% (8/190) for outdoors. In Kisian, the sporozoite rate was 1% (1/112) for indoor resting An. gambiae. None of the outdoor collected mosquitoes in Kisian tested positive for sporozoite infections (n = 73). The study reports high indoor resting densities of An. gambiae and An. funestus, insecticide resistance, and persistence of malaria transmission indoors regardless of the use of long-lasting insecticidal nets (LLINs). These findings underline the difficulties of controlling malaria vectors resting and biting indoors using the current interventions. Supplemental vector control tools and implementation of sustainable insecticide resistance management strategies are needed in western Kenya.

Evaluation of the performance of new sticky pots for outdoor resting malaria vector surveillance in western Kenya

BackgroundSurveillance of outdoor resting malaria vector populations is crucial to monitor possible changes in vector resting and feeding behaviour following the widespread use of indoor-based vector control interventions. However, it is seldom included in the routine vector surveillance system in Africa due to lack of well standardized and efficient traps. This study was conducted to evaluate the performance of sticky pots for outdoor resting malaria vector surveillance in western Kenya.MethodsMosquito collections were conducted from September 2015 to April 2016 in Ahero and Iguhu sites, western Kenya using sticky pots, pit shelters, clay pots, exit traps, Prokopack aspirator and CDC light traps (outdoor and indoor). Species within Anopheles gambiae (s.l.) were identified using polymerase chain reaction (PCR). Enzyme-linked immunosorbent assay (ELISA) was used to determine blood meal sources of malaria vectors.ResultsA total of 23,772 mosquitoes were collected, of which 13,054 were female anophelines comprising An. gambiae (s.l.) (72.9%), An. funestus (13.2%), An. coustani (8.0%) and An. pharoensis (5.9%). Based on PCR assay (n = 672), 98.6% An. arabiensis and 1.4% An. gambiae (s.s.) constituted An. gambiae (s.l.) in Ahero, while this was 87.2% An. gambiae (s.s.) and 12.8% An. arabiensis in Iguhu. The sticky pots and pit shelters showed similar performance with regard to the relative abundance and host blood meal indices of An. gambiae (s.l.) and An. funestus. In terms of density per trap, a pit shelter caught on average 4.02 (95% CI: 3.06–5.27) times as many An. gambiae (s.l.) as a sticky pot, while a sticky pot captured 1.60 (95% CI: 1.19–2.12) times as many An. gambiae (s.l.) as a clay pot. Exit traps yielded a significantly lower number of An. gambiae (s.l.) than all other traps in Ahero, but a higher number of An. gambiae (s.l.) compared to the other outdoor traps in Iguhu. Indoor CDC light traps captured a significantly higher number of An. funestus than other traps.ConclusionsSticky pots could be a useful and complementary tool for outdoor resting malaria vector surveillance, in settings where using pit shelters is not feasible and less productive. The lower vector density in the sticky pots compared to pit shelters suggests that batches of sticky pots (i.e. four per compound) need to be deployed in order to make a direct comparison. This study also highlighted the need to concurrently undertake indoor and outdoor vector surveillance to better understand residual malaria transmission.

Impact of combining indoor residual spraying and long-lasting insecticidal nets on Anopheles arabiensis in Ethiopia: results from a cluster randomized controlled trial

BackgroundIndoor residual house spraying (IRS) and long-lasting insecticidal nets (LLINs) are the key front-line malaria vector interventions against Anopheles arabiensis, the sole primary malaria vector in Ethiopia. Universal coverage of both interventions has been promoted and there is a growing demand in combinations of interventions for malaria control and elimination. This study compared the impact on entomological outcomes of combining IRS and LLINs with either intervention alone in Adami Tullu district, south-central Ethiopia. The epidemiological outcomes were recently published on a separate paper.MethodsThis factorial, cluster-randomized, controlled trial randomized villages to four study arms: IRS + LLIN, IRS, LLIN, and control. LLINs (PermaNet 2.0) were provided free of charge. IRS with propoxur was applied before the main malaria transmission season in 2014, 2015 and 2016. Adult mosquitoes were collected in randomly selected villages in each arm using CDC light trap catch (LTC) set close to a sleeping person, pyrethrum spray catch (PSC), and artificial pit shelter (PIT), for measuring mosquito host-seeking density (HSD), indoor resting density (IRD), and outdoor resting density (ORD), respectively. Human landing catch (HLC) was performed in a sub-set of villages to monitor An. arabiensis human biting rates (HBR). Mean vector densities and HBR were compared among study arms using incidence rate ratio (IRR) calculated by negative binomial regression.ResultsThere were no significant differences in mean densities (HSD, IRD, ORD) and HBR of An. arabiensis between the IRS + LLIN arm and the IRS arm (p > 0.05). However, mean HSD, IRD, ORD, and HBR were significantly lower in the IRS + LLIN arm than in the LLIN alone arm (p < 0.05). All An. arabiensis tested for malaria infection were negative for Plasmodium species. For this reason, the entomological inoculation rate could not be determined.ConclusionsThe IRS + LLIN were as effective as IRS alone in reducing densities and HBR of An. arabiensis. However, the effectiveness of the two interventions combined was higher than LLINs alone in reducing densities and HBR of the vector. Added impact of the combination intervention against malaria infectivity rates of An. arabiensis compared to either intervention alone remains unknown and warrants further research.Trial registration PACTR201411000882128. Registered 8 September 2014, https://trialsjournal.biomedcentral.com/articles/10.1186/s13063-016-1154-2

Species composition, blood meal hosts and Plasmodium infection rates of Anopheles mosquitoes in Ghibe River Basin, southwestern Ethiopia

BackgroundVector control interventions using long-lasting insecticidal nets (LLINs) and indoor residual spraying (IRS) are commonly practiced tools for the control of malaria in Ethiopia. In order to evaluate the effectiveness of these control interventions, and understand the prevailing malaria vectors, their incrimination in disease transmission, and their resting and feeding behavior, we set out to identify the Anopheles species, their blood meal sources, and entomological inoculation rate (EIR) in Ghibe and Darge within the Ghibe River basin, southwestern Ethiopia.MethodsAdult Anopheles mosquitoes were sampled both indoors and outdoors from January 2015 to October 2016 using Centers for Disease Control and Prevention (CDC) light traps, pyrethrum spray catch (PSC), artificial pit shelters and mouth aspirators. Mosquito species were morphologically identified, and their blood meal sources and malaria sporozoite rates were assessed using enzyme-linked immunosorbent assays.ResultsIn total, 13 species of Anopheles mosquitoes were identified, among which Anopheles gambiae (s.l.) was the predominant species: 87.9 and 67.7% in Ghibe and Darge, respectively. The mean density of An. gambiae (s.l.) collected per night using CDC light traps was 1.8 and 0.7 outdoors and indoors, respectively, in Ghibe, and 0.125 and 0.07 indoors and outdoors, respectively, in Darge. Anopheles mosquito abundance was higher in houses near the river than in houses far from the river in both study sites. Among Anopheles mosquitoes sampled using CDC light trap catches, 67.6% were unfed and the indoor and outdoor human blood indices of An. gambiae (s.l.) were 58.4 and 15.8%, respectively in Ghibe, while in Darge, they were 57.1 and 50%, respectively. Sporozoite rates were 0.07% for P. vivax and 0.07% for P. falciparum in Ghibe and zero in Darge. In Ghibe, the overall EIRs for P. falciparum and P. vivax were zero and 8.4 infective bites/person/year, respectively, in 2015, while zero and 5.4 infective bites/person/year for P. vivax and P. falciparum, respectively, in 2016. No Plasmodium-positive Anopheles mosquitoes were identified from Darge.ConclusionsAnopheles gambiae (s.l.), the principal vector of malaria in Ethiopia was the most abundant species both indoors and outdoors, fed both on human and cattle blood and occurred at higher frequencies near rivers. Anopheles gambiae (s.l.) that were circumsporozoite-positive for Plasmodium species were collected from Ghibe, but not Darge.

Anopheline species composition and the 1014F-genotype in different ecological settings of Burkina Faso in relation to malaria transmission

BackgroundA three-year longitudinal study was conducted in four sentinel sites from different ecological settings in Burkina Faso, between 2008 and 2010 to identify longitudinal changes in insecticide resistance within Anopheles gambiae complex species based on larval collection. During this study, adult mosquitoes were also collected indoor and outdoor using several methods of collection. The present study reports the diversity of malaria vectors and the 1014F-genotype from this adult collection and investigates the association between this 1014F-genotype and sporozoite rate.MethodsAdult mosquitoes were collected from July to August (corresponding to the start of rainy season) and October to November (corresponding to the end of rainy season) over 3 years (2008–2010) at four sites across the country, using pyrethrum spray catches (PSC), exit traps and pit shelters. Anopheles gambiae complex mosquitoes were identified to species and genotyped for the L1014F kdr mutation by PCR using genomic DNA. The circumsporozoite antigen of Plasmodium falciparum was detected in mosquitoes using sandwich ELISA.ResultsOverall 9212 anopheline mosquitoes were collected during the study period. Of those, 6767 mosquitoes were identified as Anopheles gambiae sensu lato (s.l.). Anopheles arabiensis, Anopheles coluzzii, Anopheles gambiae and or Anopheles funestus were incriminated as vectors of P. falciparum in the study area with an average sporozoite rate of 5%, (95% CI 4.14–5.99%). The kdr1014F-genotype frequencies were 11.44% (95% CI 2.5–39.85%), 19.2% (95% CI 4.53–53.73%) and 89.9 (95% CI 63.14–97.45%), respectively for An. arabiensis, An. coluzzii and An. gambiae. The proportion of the 1014F-genotype varied between sporozoite-infected and uninfected An. gambiae s.l. group. There was no significant difference in the 1014F-genotype frequency between infected and uninfected mosquitoes.ConclusionThe current study shows the diversity of malaria vectors and significant interaction between species composition and kdr1014F-genotype in An. gambiae complex mosquitoes from Burkina Faso. In this study, no associations were found between the 1014F-genotype and P. falciparum infection in the major malaria vector An. gambiae s.l.

Bionomics of Anopheles subpictus (Diptera: Culicidae) in a Malaria Endemic Area, Southeastern Iran.

Anopheles subpictus Grassi is considered a secondary malaria vector in parts of Asia. The current study determined some ecological and bionomical characteristics of this species in southeastern Iran. The temporal patterns of abundance, resting behavior, blood feeding activity, host selection, adult susceptibility to insecticides and larval habitats were investigated. Most adults were collected by pyrethrum space-spray collection, followed by pit shelters and outlet window traps, respectively. The abdominal condition index of gravid to blood fed females resting outdoors was more than one, thereby showing exophilic resting behavior. Only 25% of engorged females tested positive for human blood, even though most of the samples were collected from houses. The host seeking activity of An. subpictus was bimodal with peaks at 22-2300 h and 03-0400 h. Also, the relative abundance showed peaks in March and December. The results of susceptibility tests showed a resistance of field strains to DDT. Future studies are needed to investigate the possible role of this species in malaria transmission in southeastern Iran.

Study on the species composition and ecology of anophelines in Addis Zemen, South Gondar, Ethiopia

BackgroundMalaria is a public health problem in Ethiopia and its transmission is generally unstable and seasonal. For the selection of the most appropriate vector control measures, knowledge on the ecology of the vector is necessary at a local level. Therefore, the objectives of this study were to document the species composition, breeding habitat characteristics and occurrence of anopheline larva in Sheni stream and the vectorial role of the prevailing Anopheles in relation to malaria transmission in Addis Zemen, Ethiopia.MethodsImmature anophelines were sampled from breeding habitats and characteristics, such as water temperature, turbidity, water current, water pH and other variables, of the habitats were measured from October 2011 to February 2012. Adult anophelines were sampled inside human dwellings using space spray and Center for Disease Control light traps. Artificial pit shelters and clay pots were also used for outdoor adult collections. Anophelines collected were identified using morphological key. The enzyme-linked immunosorbent assay was applied to detect circumsporozoite proteins of Plasmodium and source of blood meals.ResultsA total of 6258 Anopheles larvae were collected and identified morphologically. Five anopheline species were found: An. gambiae (s.l.), An. cinereus, An. demeilloni, An. christi and An. pretoriensis. Anopheles gambiae (s.l.) existed in most of the habitats investigated. Only the former three species were captured in the adult collections. Sun-lit Sheni stream, rain pools, hoof prints, drainage and irrigation canals were found to be habitats of larvae. Anopheles gambiae (s.l.) larvae were most abundantly sampled from sand mining and natural sand pools of Sheni stream. Multiple regression analysis showed that clear, permanent and temporary habitats devoid of mats of algae were the best predictors of An. gambiae (s.l.) larval abundance. It is also the responsible malaria vector in the study area and exhibits anthropophilic and endophagic behaviour.ConclusionsThe malaria vector An. gambiae (s.l.) was found in Addis Zemen throughout the study period from both adult and larval collections. Sheni stream is the main larval habitat responsible for the occurrence of anopheline larvae during the dry season of the study area when other breeding sites perish.

Indoor and outdoor malaria vector surveillance in western Kenya: implications for better understanding of residual transmission

BackgroundThe widespread use of indoor-based malaria vector control interventions has been shown to alter the behaviour of vectors in Africa. There is an increasing concern that such changes could sustain residual transmission. This study was conducted to assess vector species composition, feeding behaviour and their contribution to indoor and outdoor malaria transmission in western Kenya.MethodsAnopheles mosquito collections were carried out from September 2015 to April 2016 in Ahero and Iguhu sites, western Kenya using CDC light traps (indoor and outdoor), pyrethrum spray catches (PSCs) (indoor) and pit shelters (outdoor). Species within Anopheles gambiae s.l. and Anopheles funestus s.l. were identified using polymerase chain reaction (PCR). Enzyme-linked immunosorbent assay (ELISA) was used to determine mosquito blood meal sources and sporozoite infections.ResultsA total of 10,864 female Anopheles mosquitoes comprising An. gambiae s.l. (71.4%), An. funestus s.l. (12.3%), Anopheles coustani (9.2%) and Anopheles pharoensis (7.1%) were collected. The majority (61.8%) of the anopheline mosquitoes were collected outdoors. PCR result (n = 581) revealed that 98.9% An. arabiensis and 1.1% An. gambiae s.s. constituted An. gambiae s.l. in Ahero while this was 87% An. gambiae s.s. and 13% An. arabiensis in Iguhu. Of the 108 An. funestus s.l. analysed by PCR, 98.1% belonged to An. funestus s.s. and 1.9% to Anopheles leesoni. The human blood index (HBI) and bovine blood index (BBI) of An. arabiensis was 2.5 and 73.1%, respectively. Anopheles gambiae s.s. had HBI and BBI of 50 and 28%, respectively. The HBI and BBI of An. funestus was 60 and 22.3%, respectively. Forage ratio estimate revealed that An. arabiensis preferred to feed on cattle, An. gambiae s.s. showed preference for both human and cattle, while An. funestus preferred human over other hosts. In Ahero, the sporozoite rates for An. arabiensis and An. funestus were 0.16 and 1.8%, respectively, whereas in Iguhu, the sporozoite rates for An. gambiae s.s. and An. funestus were 2.3 and 2.4%, respectively. In Ahero, the estimated indoor and outdoor entomological inoculation rate (EIR) was 108.6 infective bites/person/year (79.0 from An. funestus and 29.6 from An. arabiensis) and 43.5 infective bites/person/year (27.9 from An. arabiensis and 15.6 from An. funestus), respectively. In Iguhu, the estimated indoor and outdoor EIR was 24.5 infective bites/person/year (18.8 from An. gambiae s.s. and 5.7 from An. funestus) and 5.5 infective bites/person/year (all from An. gambiae s.s.), respectively.ConclusionAnopheles gambiae s.s. showed an increasing tendency to feed on cattle. Anopheles arabiensis was highly zoophagic, whereas An. funestus showed anthropophagic behaviour. While the majority of malaria transmission occurred indoor, the magnitude of outdoor transmission was considerably high. Additional control tools that complement the existing interventions are required to control residual transmission.

Fauna and some biological characteristics of Anopheles mosquitoes (Diptera: Culicidae) in Kalaleh County, Golestan Province, northeast of Iran

ObjectiveTo determine fauna and some ecological aspects of Anopheles mosquitoes in northeast of Iran. MethodsIn this descriptive study, 3 villages in Kalaleh County were selected in different geographical zones. Anopheles mosquitoes were collected biweekly from May to October using standard dipping method for larvae, and hand catch, total catch, artificial pit shelter as well as night-biting collections on human and animal baits for adults. ResultsTotally 399 larvae and 2602 adults of Anopheles mosquitoes were collected and identified as 2 species: Anopheles superpictus s.l. (An. superpictus s.l.) and Anopheles maculipennis s.l. The dominant species was An. superpictus s.l. (92.1%). Activity of these mosquitoes found to be started from middle of May and extended till September with two peaks of activity in July and August. ConclusionsAn. superpictus s.l. as one of the main malaria vectors in Iran as well as some other parts of the world is the dominant species in the study area. This species has high potential for transmission and possibility of establishing a transmission cycle with low abundance. Other species, Anopheles maculipennis s.l. also has introduced as a malaria vector in northern parts of Iran. As this Anopheles is a complex species, genetic studies are recommended to determine the members of this complex in the study area.

Insights Into Resting Behavior of Malaria Vector Mosquitoes in Mutare and Mutasa Districts of Manicaland Province, Zimbabwe.

A study was conducted to investigate the current resting behavior of malaria vectors in Mutare and Mutasa districts, Zimbabwe. Mosquitoes were captured using pyrethrum spray collection, prokopac aspirator, pit shelter, and exit trap methods. Mosquitoes were sorted and identified using morphological key and polymerase chain reaction (PCR) techniques. The Anopheles funestus group constituted 97%, whereas Anopheles gambiae complex mosquitoes were few (3%). Endophilic collections in both species were five times greater than exophilic catches. The endophilic trait was further demonstrated by gravid to fed index (gravid/fed) of constantly more than 1. Nearly 90% endophilic An. funestus populations were collected on sprayable and 10% collected on unsprayable surfaces. Of the sprayable surfaces, 56% were collected on the roofs, with 44% on the walls. Of those on the walls, 44, 22, and 34% were caught on wall heights >1, 1.0-1.5, <1.5 m from the ground, respectively. Of the gravid An. funestus caught, nearly two-thirds were collected exiting pyrethroid-treated structures, with a 24-h mortality of less than 10%. The PCR analysis of 120 specimens taken randomly from the An. funestus group was all An. funestus s.s. The present work indicates that for effective malaria control in Mutare and Mutasa districts using indoor residual spraying, both walls and roofs must be sprayed.

Malaria incidence and entomological findings in an area targeted for a cluster-randomized controlled trial to prevent malaria in Ethiopia: results from a pilot study.

BackgroundThis study was part of the work to prepare for a cluster-randomized controlled trial to evaluate the effect of combining indoor residual spraying and long-lasting insecticidal nets on malaria incidence. A pilot study was done to estimate the variations of malaria incidence among villages, combined with entomological collections and an assessment of susceptibility to insecticides in malaria vectors.MethodsA cohort of 5309 residents from four kebeles (the lowest government administrative unit) in 996 households was followed from August to December 2013 in south-central Ethiopia. Blood samples were collected by a finger prick for a microscopic examination of malaria infections. A multilevel mixed effect model was applied to measure the predictors of malaria episode. Adult mosquitoes were collected using light traps set indoors close to a sleeping person, pyrethrum spray sheet catches and artificial outdoor pit shelters. Enzyme-linked immunosorbent assays were used to detect the sources of mosquito blood meals, while mosquito longevity was estimated based on parity. The World Health Organization’s tube bioassay test was used to assess the insecticide susceptibility status of malaria vectors to pyrethroids and carbamates.ResultsThe average incidence of malaria episode was 4.6 per 10,000 person weeks of observation. The age group from 5 to 14 years (IRR = 2.7; 95 % CI 1.1–6.6) and kebeles near a lake or river (IRR = 14.2, 95 % CI 3.1–64) were significantly associated with malaria episode. Only 271 (27.3 %) of the households owned insecticide-treated nets. Of 232 adult Anopheles mosquitoes collected, Anopheles arabiensis (71.1 %) was the predominant species. The average longevity of An. arabiensis was 14 days (range: 7–25 human blood index days). The overall human blood index (0.69) for An. arabiensis was higher than the bovine blood index (0.38). Statistically significant differences in Anopheline mosquitoes abundance were observed between the kebeles (P = 0.001). Anopheles arabiensis was susceptible to propoxur, but resistant to pyrethroids. However, An. pharoensis was susceptible to all pyrethroids and carbamates tested.ConclusionsThis study showed a high variation in malaria incidence and Anopheles between kebeles. The observed susceptibility of the malaria vectors to propoxur warrants using this insecticide for indoor residual spraying, and the results from this study will be used as a baseline for the trial.

Zoophagic behaviour of anopheline mosquitoes in southwest Ethiopia: opportunity for malaria vector control.

BackgroundIncreased understanding of the feeding behaviours of malaria vectors is important to determine the frequency of human-vector contact and to implement effective vector control interventions. Here we assess the relative feeding preferences of Anopheles mosquitoes in relation to cattle and human host abundance in southwest Ethiopia.MethodsWe collected female Anopheles mosquitoes bi-weekly using Centers for Disease Control and prevention (CDC) light traps, pyrethrum spray catches (PSCs) and by aspirating from artificial pit shelters, and determined mosquito blood meal origins using a direct enzyme-linked immunosorbent assay (ELISA).ResultsBoth Anopheles arabiensis Patton and An. marshalli (Theobald) showed preference of bovine blood meal over humans regardless of higher human population sizes. The relative feeding preference of An. arabiensis on bovine blood meal was 4.7 times higher than that of human blood. Anopheles marshalli was 6 times more likely to feed on bovine blood meal than humans. The HBI of An. arabiensis and An. marshalli significantly varied between the collection methods, whereas the bovine feeding patterns was not substantially influenced by collection methods. Even though the highest HBI of An. arabiensis and An. marshalli was from indoor CDC traps collections, a substantial number of An. arabiensis (65 %) and An. marshalli (63 %) had contact with cattle. Anopheles arabiensis (44 %) and An. marshalli (41 %) had clearly taken bovine blood meals outdoors, but they rested indoors.ConclusionAnopheles mosquitoes are zoophagic and mainly feed on bovine blood meals than humans. Hence, it is important to consider treatment of cattle with appropriate insecticide to control the zoophagic malaria vectors in southwest Ethiopia. Systemic insecticides like ivermectin and its member eprinomectin could be investigated to control the pyrethroid insecticides resistant vectors.