- Open Access
Establishment of a large semi-field system for experimental study of African malaria vector ecology and control in Tanzania
© Ferguson et al; licensee BioMed Central Ltd. 2008
Received: 13 May 2008
Accepted: 20 August 2008
Published: 20 August 2008
Medical entomologists increasingly recognize that the ability to make inferences between laboratory experiments of vector biology and epidemiological trends observed in the field is hindered by a conceptual and methodological gap occurring between these approaches which prevents hypothesis-driven empirical research from being conducted on relatively large and environmentally realistic scales. The development of Semi-Field Systems (SFS) has been proposed as the best mechanism for bridging this gap. Semi-field systems are defined as enclosed environments, ideally situated within the natural ecosystem of a target disease vector and exposed to ambient environmental conditions, in which all features necessary for its life cycle completion are present. Although the value of SFS as a research tool for malaria vector biology is gaining recognition, only a few such facilities exist worldwide and are relatively small in size (< 100 m2).
The establishment of a 625 m2 state-of-the-art SFS for large-scale experimentation on anopheline mosquito ecology and control within a rural area of southern Tanzania, where malaria transmission intensities are amongst the highest ever recorded, is described.
A greenhouse frame with walls of mosquito netting and a polyethylene roof was mounted on a raised concrete platform at the Ifakara Health Institute. The interior of the SFS was divided into four separate work areas that have been set up for a variety of research activities including mass-rearing for African malaria vectors under natural conditions, high throughput evaluation of novel mosquito control and trapping techniques, short-term assays of host-seeking behaviour and olfaction, and longer-term experimental investigation of anopheline population dynamics and gene flow within a contained environment that simulates a local village domestic setting.
The SFS at Ifakara was completed and ready for use in under two years. Preliminary observations indicate that realistic and repeatable observations of anopheline behaviour are obtainable within the SFS, and that habitat and climatic features representative of field conditions can be simulated within it. As work begins in the SFS in Ifakara and others around the world, the major opportunities and challenges to the successful application of this tool for malaria vector research and control are discussed.
Recent advances in genomics and bioinformatics are allowing science to test the boundaries of reductionism as never before: how well can biological processes observed at the level of individual molecules, genes or cells predict the behaviour of more complex systems such as whole organs, individuals, or populations? These exciting technological developments have generated renewed interest within existing, more long-established biological disciplines to seek out empirical tools for quantifying and testing the relationship between phenomena occurring at different levels of biological organization in order to generate better predictions. One such field is medical entomology. Typically research in medical entomology is conducted at two very different scales: the first being laboratory-based studies of arthropod disease vectors under controlled insectary conditions, and the second being large-scale epidemiological surveys of their abundance and distribution in nature. While the former approach is clearly advantageous for identifying potential biological mechanisms, and the latter for generating hypotheses from correlations, it is increasingly recognized that the ability to make inferences across these scales is hindered by a conceptual and methodological gap in between that limits our ability to conduct hypothesis-driven empirical research on relatively large and environmentally realistic scales .
Arguably for the first time in the history of their discipline, medical entomologists now find themselves in the unique position of having both the need for such experimental initiatives recognized , and the financial support to create them becoming available through new funding streams in global health. The need to fill the research gap between laboratory and field is also stimulated by awareness that although current approaches to important vector-borne diseases such as malaria based on artemisinin-based combination therapies and insecticide treated bed nets are proving successful [3–7], their long-term effectiveness may be undermined by the emergence of drug and insecticide resistance [8–10]. Consequently, the need for new strategies that exploit novel aspects of vector genetics, physiology, behaviour and ecology are increasingly needed. These innovations must be drawn from an understanding of vector biology within natural transmission settings if they are to yield rapid, locally appropriate strategies for disease control.
While almost all new approaches to vector control could benefit from a closer integration of laboratory and field perspectives , the most prominent candidate is the development of transgenic parasite resistant and/or sterile vectors whose release into the wild could reduce disease transmission by reducing parasite and/or vector populations [12–15]. In the past decade, the genetic transformation of a number of important disease-transmitting mosquito species has become possible [16–20]. Transgenes have been identified that deliver effector molecules that substantially reduce the development of rodent malaria parasites [21–23] and human dengue virus within mosquitoes , which has fuelled optimism that mass-release of laboratory-reared genetically-modified individuals could reduce disease transmission. The greatest unknown with respect to the feasibility of this approach is whether genetically-modified mosquitoes would be able to survive and successfully compete for mates against their wild counterparts outside of the confines of the laboratory. Initial laboratory studies indicated that transgenes impose fitness costs which reduce the reproductive success of the mosquito bearer [25–27]. A recent study suggests this disadvantage can be reduced by use of out-crossed mosquito lines , although so far only under conditions where exposure to parasites is substantially greater than mosquitoes encounter in the wild. While this improvement is encouraging, it does not address the problem that all laboratory-reared mosquitoes, regardless of their genotype, may have poor competitive ability in the wild. For example, recent comparative analysis of Anopheles gambiae s.s. in captivity and in nature in southern Tanzania suggest free-living males are larger and have greater lipid reserves than those reared under apparently optimal laboratory conditions . Regardless of whether this reduction in energetic reserves was due to selection for smaller individuals during the colonization process and/or sub-optimal conditions of insectary environments, it suggests laboratory-reared mosquitoes could be at a sizeable disadvantage to their wild counterparts.
Additional studies have shown that the mating success of male mosquitoes depends on subtle variation in environmental conditions experienced during larval development [30, 31], which may not be fully captured in mass-rearing facilities. These limitations are thought to have been responsible for the failure of many genetic control trials during the 1970's and 80's, which found that laboratory-reared male mosquitoes were unable to compete in the wild . Clearly, to avoid repeating these failures with the new generation of transgenic mosquitoes, intermediary testing grounds between the laboratory and field within disease-endemic countries are needed.
Semi-Field Systems (SFS) have been proposed as the best mechanism for bridging this gap. A semi-field system is here defined as an enclosed environment, ideally situated within the natural ecosystem of the target disease vector and exposed to ambient environmental conditions, within which all features necessary for its lifecycle completion are present . In the case of mosquito vectors of human disease, this typically involves a large outdoor cage in which the movement of the disease vector of interest either in or out of the unit is restricted by netting, and within which features such as aquatic larval habitats, blood hosts for adult females, sugar sources (plants) for adults, appropriate resting sites (houses, cattle sheds, etc.) and environmental features (e.g swarm markers to stimulate mating), are present. There are no general guidelines for the appropriate size of such a unit, but ideally it should be large enough to sustain a population of similar density to that encountered in the target environment for numerous generations.
This definition of a SFS differs from others that apply 'semi-field' to studies that actually involve observation of vectors in a non-contained setting or habitat, where only one part of its life cycle is present . A major goal of SFS is to establish multiple generations of a vector population within a contained setting, without outside intervention  in addition to facilitating short-term behavioural or ecological studies based on a single cohort. The main advantage of this approach is that because the abundance and composition of vectors within the SFS can be known, and if desired experimentally manipulated (either at the time of introduction, or through removal of some target individuals), much more precise estimates of the value and variability of demographic and life-history parameters can be obtained than would be from the field. Additionally, they allow researchers to conduct high throughput assays of control tools and ecological phenomena year round without risk of exposure to infection, as all mosquitoes used within the SFS will be free of parasites.
Previous and current location, size, target species and research aims of Semi-Field Systems (SFS) established for mosquito vector research.
Number of units
10 × 5 × 6
Various European anophelines
Basic ecological studies
India – Madras
12.2 6.1 × 3.05
Basic ecological studies, evaluation of genetic control strategies for population suppression
India – Delhi
5.6 × 3.3 × 2.1
Ae. aegypti Cx. Fatigans
11.4 × 7.1 × 4.4
An. gambiae s.s.
Basic ecological studies, vector-malaria parasite interactions, evaluation of novel trap designs and repellents
10 × 10 × 4
Basic ecological studies
Tanzania – Muheza
12.2 × 8.2 × 4.6
An. gambiae s.s Cx. quinquefasciatus
Evaluation of trapping methods, training and basic ecological studies
18 × 8 × 2.75
Fitness of sterilized males, basic ecological studies
Tanzania – Ifakara
29.8 × 21 × 7.1
An. gambiae s.s An. arabiensis
Basic ecological studies, evaluation of trapping methods and repellents
17 × 9 × 4.3
Assessment of biocontrol strategy using Wolbachia, basic ecological studies
25 × 10 × 3
Research on Sterile Insect Technique
Materials and methods
The SFS was estabished at the Ifakara Health Institute (IHI) located in the Kilombero district of southern Tanzania. Malaria transmission intensities within this area are amongst the highest described for sub-saharan Africa [49, 50]; with annual entomological inoculation rates exceeding three hundred infectious bites a year in some locations [49, 51, 52]. The major malaria vectors in this region are Anopheles arabiensis, An. gambiae s.s. and An. funestus [52–54].
SFS site selection
The crucial first step in establishing a SFS is identifying an appropriate site that adequately captures the environmental conditions experienced by local mosquito species. Additional logistic criteria include ease of access by research personnel and electricity/water supply, being situated where potential hazards to surrounding residents arising from accidental vector release are negligible, and continual monitoring by security staff is possible. Trade-offs may arise in attempting to maximize all these criteria at particular locations which will require careful case-by-case consideration. For example, it has been suggested that the best way to limit hazards posed by unintentional release of mosquitoes into the environment would be to build containment units as far away from communities as possible . However, the majority of SFS currently in existence and being planned are located within disease-endemic settings in the developing world. In many of these settings, access to roads, water, an electrical supply, and reliable 24-hour surveillance is possible only near towns or cities. In balancing these components of potential risk, it was decided to select a site for the SFS that is within the campus of the IHI, which is located in Ifakara town. By building within the fenced-off perimeter of the research centre, it was possible to ensure constant surveillance and containment, and strictly control those who had access to the SFS.
Another key factor in the site selection process for SFS is the availability of background data on the dynamics of local vector populations and their disease transmission ability . This information is essential to examine how closely the behaviour, life-history and population dynamics of contained vectors represent those of the wild. As mosquitoes in the SFS will be exposed to many of the same environmental conditions as those of neighbouring populations (e.g temperature, humidity, vegetation), it is anticipated they will be subject to similar selective forces. However, one deviation from complete 'naturalness' was made in the IHI SFS by covering its roof with polyethylene plastic; a decision taken on the basis that this compromise would permit experimental manipulation of rainfall in future experiments. How this modification influences the environmental suitability of the SFS relative to ambient conditions can be assessed by comparison of mosquito population dynamics in the SFS with those of the surrounding area. An advantage of selecting a site in Ifakara was that substantial baseline epidemiological and entomological information on the dynamics of malaria and Anopheles populations in the area is already available [50, 54, 56, 57]. Additionally, detailed knowledge of mosquito ecology exists for the Kilombero valley, and new studies specifically addressing the mating biology [29–31, 58] and population genetics (Ng'habi et al., in prep.) of An. gambiae and An. arabiensis within this region were initiated concurrently with the establishment of the SFS.
Planning and design
Given that Ifakara town is occasionally subject to flooding during the rainy reason, it was decided that the entire SFS structure should be raised 1.6 m above ground level to ensure floodwaters would not breach the structure even during heavy precipitation. The SFS was thus mounted on top of a 22 × 30 m steel-reinforced concrete platform of 0.16 m thickness. This platform was supported by 56 steel-reinforced concrete posts (1.1 m × 1.1 m) equidistantly spaced along the length and width which would allow for natural water flow to continue unimpeded under the structure during times of heavy floods.
Ethical considerations and community awareness
A potential risk of using SFS in disease-endemic settings is the accidental release of vectors into an environment where they could become infected with a human pathogen, increase the size of the local vector population or introduce a novel phenotype with enhanced transmission capacity. Consequently great care and vigilance is required to ensure the physical integrity of the structure and containment protocols. Access to the IHI SFS is restricted to a small number of research personnel. Research technicians conduct weekly intensive inspections of all areas of the inner and outer structure for physical damage that could allow mosquitoes to escape or enter from outside.
In addition to making sure that mosquitoes do not escape from a SFS, it is also imperative to ensure that malaria parasites are not accidentally introduced through mosquito contact with an infected person. A protocol for weekly malaria screening for all those working within the SFS was developed. Individuals found to be infected during this screening would be immediately treated with appropriate first line anti-malarial medication and excluded from the screen-houses for one month. Should it be found that a staff member has had malaria parasites while working within the SFS, the experimental chamber in which they worked can be shut down and all mosquitoes within it killed (by depriving them of water, blood and breeding sites for at least two weeks) to ensure no potentially infectious mosquitoes remain within it. Additional methods to reduce the risk of the unintentional introduction of parasites include the use of non-amplifying animal hosts such as livestock as the main blood source for captive vector populations. This procedure is being adopted in the IHI SFS where cows are used as the blood source for free-living Anopheles populations.
In addition to the precautions described above, a key ethical requirement of working with SFS is the creation and maintenance of strong support and awareness within the local community for these research activities. A series of public meetings with IHI staff, workers involved with the construction of the SFS, district health and government officials, and local residents were held in which information on the function and purpose of the SFS was disseminated. Ethical clearance from both the IHI Institutional Review Board (IHDRC/EC4/CL.N96/2004) and Tanzanian National Institute of Medical Research (NIMR/HQ/R.8a/Vol.IX/345) for SFS studies was obtained before the start of this study.
Constructing the SFS
Establishing research activities
Different research activities were allocated to each of the four SFS sections on the basis of maximizing logistical efficiency and minimizing the risk of mosquito escape or entry from outside. The first section behind the main front access point (section 1, Figure 3b) was designated for use as an An. arabiensis insectary. All mosquitoes in this section are thus additionally contained either in adult cages, or larval trays covered with netting. Initial consultation with the greenhouse manufacturer suggested that the netted outer walls would allow temperatures inside the SFS to equilibrate with ambient conditions outside. Ambient temperatures during the hot rainy season in Ifakara can exceed 40°C for several hours each day which are sufficiently high to kill adult and larval mosquitoes. To buffer the insectary from extreme temperatures that could knock out the colony, a traditional thatch roof was built within the insectary area to provide additional shading and cooling (Figure 3b). Under these conditions, an average of 638 pupae per day (± 114.8) were obtained from the F1 generation of An. arabiensis collected from a nearby village in March 2008.
The insectary connects directly onto two experimental spaces; the first being a 9.6 m × 10 m chamber within which an experimental hut (3.5 m × 4 × 2.5 m) was constructed for studies of mosquito host seeking and house entry behaviour (Figure 3c). This hut was fitted with 6 window traps that can be used to capture mosquitoes leaving and/or attempting to enter the house while a live host is within it . This experimental hut section is designated for short-term behavioural studies in which no more than 300 mosquitoes at a time are released (at dusk), and subsequently recaptured the next day and removed in a cage. A further screen door separates this section from the insectary area meaning that three security doors must be passed through before reaching the outside, and minimizing the risk of mosquito escape during exit or entry.
The insectary also connects directly to a 9.1 × 21 m chamber designated for establishment of a free-living, self-replicating An. arabiensis population within a realistic ecosystem (Figure 3d). This section is intended for study of Anopheles behaviour, ecology and gene flow within an environment that mimics the natural surroundings as closely as possible. The exact number of free-living mosquitoes that will be held within this unit is uncertain, and will depend upon the carrying capacity of the established population at equilibrium. This section is linked to the main insectary by another double entry door system, requiring four doors to be passed through before reaching outside.
The fourth experimental section (9.1 × 21 m) is set up as an stand-alone experimental unit isolated from all other areas of the SFS, within which studies of olfaction and chemical ecology are ongoing (Figure 3e). This section is physically separated from the adjoining central section by thick polyethylene plastic which minimizes the direct flow of air and odours between them. Entry into this section is possible only from the rear SFS double entry door, and not through any other adjoining section. Studies using odour-baited traps to compare the attraction and repellency of different compounds to Anopheles gambiae s.s. are being conducted in this section.
Replicating the natural environment
Average temperatures at different locations within the SFS from February 29th – May 9th 2008.
Ground-level in SFS
Inside mud house
23.86 – 34.69
Artificial breeding site
25.19 – 36.67
22.60 – 34.43
In just under two years a 625 m2 state-of-the-art SFS for large-scale experimentation on anopheline mosquito ecology and control was established within a remote area of southern Tanzania where malaria transmission intensities are amongst the highest ever recorded [49–52, 68]. This unique facility is more than 4 times larger than any SFS previously or currently in existence, and has capacity for a wide variety of research activities including mass-rearing of African malaria vectors under natural conditions, high throughput evaluation of novel control and trapping techniques, short-term assays of host-seeking behaviour and olfaction, and long-term experimental investigation of anopheline population dynamics and gene flow within a contained environment that simulates a local village domestic compound. This was accomplished through a multidisciplinary collaboration between entomologists, senior public health scientists of the IHI, architects, engineers, site managers and a dedicated team of labourers who built this structure largely in the absence of electricity or any other mechanized construction aids.
Experimental activities have only recently been initiated within the SFS, and the ultimate value of this facility as a research tool will be realized as the studies now underway reach conclusion. Preliminary results from short-term behavioural assays of An. gambiae host-seeking behaviour using odour-baited traps and live animal baits suggest that realistic and repeatable results can be obtained within the SFS in a relatively short period of time (I. Lyimo & S. Moore, pers. comm.). The longer-term task of establishing a self-replicating, free-living population of An. arabiensis within simulated village conditions is currently underway. Although too early to forecast the outcome of this objective, the fact that an amenable spectrum of climatic conditions can be generated within the SFS is encouraging. Although daily temperatures within exposed areas of the SFS routinely exceeded the optimum temperature of An. gambiae [26.5 C under insectary conditions, ] for some periods of the day, they were not significantly higher than those of the natural environment immediately outside of the SFS. Had the aim been to create an insectary facility for efficient mass production of An. gambiae, the regular daily periods of excessive ambient temperatures within the SFS (>35°C) would be a cause for concern. However the goal was instead to simulate the ambient climatic conditions within the Kilombero region and this was accomplished. Furthermore, the features that were built within the SFS provided microclimatic refuges in which mean temperature and variability was substantially reduced and stayed within the acceptable limits for adult survival and reproduction. For example, the mean temperature within the mud house inside the SFS was 3.5°C lower than exposed areas of the SFS. This observation matches reports from South Africa of air temperature inside mud and thatch houses being 3–6°C cooler than ambient conditions . Temperatures within the mud house in our SFS were significantly higher than those reported in a similar structure within the SFS at Mbita, western Kenya , which may be more reflective of the different climatic conditions between study sites than structural differences in SFS design. Water temperatures within the artificial larval habitats in our SFS were higher than the reported optimal value gauged from insectary studies [24–26 C, ] and those reported for the Mbita SFS , but remained within the natural range observed in An. gambiae s.l. larval habitats in east Africa [72, 73]. Given that free-living An. gambiae s.s. were able to complete their life cycle within the slightly cooler and much smaller confines of the Mbita SFS  there is optimism that the same can be achieved in the IHI SFS with An. arabiensis, a species known to have greater tolerance of hot and arid environments [66, 74].
While early observations are promising, much still remains to be known about how representative conditions inside the SFS will be of mosquito ecology in the wild. Open questions include whether a self-replicating population can be maintained over numerous generations on this spatial scale, what carrying capacity this population will reach under ambient climatic and host (bovine) conditions, whether additional climatic refugia or controls will be needed, and if existing plant and nectar sources within the SFS will be sufficient to maintain the adult male population. Importantly, the identification of limitations in the ability of our SFS to replicate natural mosquito dynamics as experimental work progresses will in itself provide valuable knowledge of the crucial determinants of anopheline population growth and stability that would not be possible under natural field conditions.
As research begins at the SFS in Ifakara and similar facilities around the world (Table 1), it is useful to consider the major challenges to the successful use of this research tool. These challenges are varied and range from the purely scientific to those of logistics and ethics. Five key areas merit discussion. The first is the possibility that although biological inferences made from SFS may be much more realistic than those from cage studies, they may still misrepresent some areas of mosquito ecology and population processes in nature. For example, although full life cycle completion of An. gambiae s.s. was achieved within the SFS in western Kenya, it was noted that the artificial breeding sites within it gave rise to considerably fewer larvae than expected . Whether this reduced efficiency was due to a problem with the environmental conditions inside the SFS, or maladaptation of the laboratory population used in these experiments to ambient conditions is unknown, but suggests there could be unique constraints or bottlenecks acting on population growth within these systems. Conversely, absence of the full range of environmental risks within the SFS such as stochasticity in host encounter rates, insecticide treated bed nets, predation by small vertebrates, pathogens and extreme environmental conditions such as flooding may result in an overestimation of life-history and demographic rates. For example, Knols et al estimated the daily survival of An. gambiae s.s within their SFS to be 90% which is higher than reported in many field studies . In order to reduce the risk of accidental parasite introduction, non-amplifying animal hosts will be used as the main source of blood in many SFS. While numerous disease vectors include the blood of non-human animals in their diet, many of the species that are most problematic exhibit a pronounced preference for humans . Several studies have shown that the fitness haematophagous insects derive from blood varies with host species [77–82]. It is also known that selection for divergent preference for human or cow hosts in An. gambiae mosquitoes can be generated in as little as 5–6 generations of selection . Thus constantly exposing vector populations within SFS to non-human hosts could result in the generation of individuals with different phenotypes, genotypes and population dynamics than those who feed on and transmit disease to humans. Continued monitoring and comparison of SFS results to those observed in the field will be useful to identify which, if any, of these issues pose serious obstacles to interpretation and reinforce the point that SFS studies are intended to complement but not replace field studies.
A second scientific concern is that vector populations established within SFS will likely be considerably smaller than those in the wild and thus experience inbreeding and a resultant reduction in genetic diversity which could impede fitness. It is well known that genetic diversity within insect vectors can be considerably reduced during laboratory colonization [84–86]. Free-living populations established within SFS may be considerably larger than typical laboratory colonies and thus avoid a similar intensity of inbreeding, however it is unlikely they will escape some bottle-necking and an associated loss of diversity from founder populations. Within only a few generations of laboratory colonization, mosquitoes can develop significant behavioural divergence from wild populations which restricts mating between them . This phenomenon may also occur within SFS, although perhaps at a slower rate than in small laboratory cages. As genetic and phenotypic divergence between contained SFS and wild populations may be unavoidable, the need for repeated comparative sampling of individuals in both settings is advocated to track if and how genetic diversity is reduced in captivity, and provide guidelines for how frequently captive populations should be enriched by fresh genetic material to maintain representative levels of diversity.
Should self-replicating vectors be successfully established in SFS, a logistical obstacle to the estimation of precise demographic rates from them will be the problem of disentangling overlapping generations. While much more precise estimates of mosquito population size will be possible within the contained environment of an SFS than in nature, it will remain difficult to accurately monitor individual-level activities such as mating behaviour and resource acquisition, and its resultant impact on fitness. The development of novel marking schemes using stable isotopes [88, 89] or distinct genetic traits may permit more precise monitoring of the behaviour and reproductive success of specific subsets of individuals, or individuals themselves.
For greatest public health relevance, SFS should be situated within or as near as possible to natural disease transmission environments as possible. Placing a large contained population of competent disease vectors within an appropriate transmission setting will always raise biosecurity concerns. Any breach of containment could result in increasing the disease transmission within the local area, and the accidental introduction of parasites into contained populations from asymptomatic carriers working within the facility could also generate the potential for infection. Awareness and discussion of how to prevent those risks are absent from early accounts of SFS use, but are justifiably coming to the forefront as plans for large-scale studies with genetically-modified disease vectors come under development. Recently an international committee of scientists formalized guidelines on recommended biosecurity measures and precautions for contained SFS trials with genetically-modified mosquitoes . The publication of these guidelines represents a significant step forward in thinking regarding the ethical responsibility for good practice within these facilities.
A final, crucial issue for the expansion of SFS-based research programmes throughout the world is the need to engage and promote awareness within the communities that host these facilities. The communities surrounding SFS research facilities should be the primary beneficiaries of research conducted within them, and their particular needs as end-users must be kept in mind when using these facilities to trial new vector control strategies. While researchers working in SFS may have this goal clearly in mind, it will be of little value unless clearly and regularly communicated to local communities in an open and discursive manner. Understandably local residents may be apprehensive about the placement of an SFS containing live insect vectors near their home, and misinformation about the purpose of this work and risks associated with it could cause considerable friction. This lesson was painfully learnt by scientists working at the Indian Council for Medical Research in the 1970's who had their research unit on Aedes mosquitoes shut down when journalists falsely alledged that the actual purpose of the work was biological warfare and human population control . That such a debacle could occur in one of the most successful disease vector control research programmes of all time [with 104 papers published in 6 years and numerous insights into population suppression gained, ] is a sobering thought for all those involved in this new generation of SFS research. Community awareness activities have begun in Ifakara, and must be sustained and scaled up if both local community members and researchers working at the SFS are to obtain maximum benefits from this research facility.
Engagement must extend beyond local communities to include scientists and students working within the disease endemic countries that host SFS. These facilities can provide substantial indirect benefits by acting as state-of-the-art training tools for young vector biologists in which methodological skills can be honed, and independent research hypotheses experimentally tested in a disease-free setting. Currently, at the IHI, there are three east African postgraduate students pursuing their PhD studies on research based within the SFS and plans to recruit several more underway. Thus this research tool will contribute to the IHI's goal of substantially increasing Ph.D-level capacity in malaria vector research within Tanzania and east Africa. Much of the recent motivation for initiating SFS programmes has been driven by laboratory-based research on genetic modification of disease vectors that has occurred almost exclusively in developing countries. For both the transgenic approach and other emerging vector control strategies to fulfill their potential, it is absolutely imperative that endemic country scientists are actively involved in driving SFS-based research and taking forward innovative techniques developed within it.
This work is dedicated to the people of Kilombero and Ulanga districts, Tanzania in the hope that the facilities described here will help bring practical solutions to reduce the unacceptable burden that malaria places upon their communities. The authors express their sincere gratitude to the TTCIH maintenance team whose commitment made this project feasible. We thank the IHI administration for their excellent guidance in project management, and all members of the Public Health Entomology team for their support. This work was supported by a VIDI grant (no. 864.03.004) awarded by the Dutch Scientific Organization (NWO) to BGJK and BBSRC David Phillips Fellowship to HMF, and the Bill and Melinda Gates Foundation (grant # GCGH121). We thank the Filclair corporation for provision of the SFS at reduced cost, and their technical support.
- Hewitt JE, Thrush SF, Dayton PK, Bonsdorff E: The effect of spatial and temporal heterogeneity on the design and analysis of empirical studies of scale-dependent systems. Am Nat. 2007, 169 (3): 398-408.View ArticlePubMedGoogle Scholar
- Clayton J: Scientists plan field tests for GM mosquitoes. Lancet Infect Dis. 2006, 6 (4): 191-192.View ArticleGoogle Scholar
- Schellenberg JR, Abdulla S, Nathan R, Mukasa O, Marchant TJ, Kikumbih N, Mushi AK, Mponda H, Minja H, Mshinda H, Tanner M, Lengeler C: Effect of large-scale social marketing of insecticide-treated nets on child survival in rural Tanzania. Lancet. 2001, 357: 1241-1247.View ArticlePubMedGoogle Scholar
- Hawley WA, Phillips-Howard PA, ter Kuile FO, Terlouw DJ, Vulule JM, Ombok M, Nahlen BL, Gimnig JE, Kariuki SK, Kolczak MS, Hightower AW: Community-wide effects of permethrin-treated bed nets on child mortality and malaria morbidity in western Kenya. Am J Trop Med Hyg. 2003, 68 ( (4 Suppl)): 121-127.PubMedGoogle Scholar
- Barnes KI, Durrheim DN, Little F, Jackson A, Mehta U, Allen E, Dlamini SS, Tsoka J, Bredenkamp B, Mthembu DJ, White NJ, Sharp BL: Effect of artemether-lumefantrine policy and improved vector control on malaria burden in KwaZulu-Natal, South Africa. PLOS Med. 2005, 2: 1123-1134.View ArticleGoogle Scholar
- Abdulla S, Gemperli A, Mukasa O, Armstrong Schellenberg JR, Lengeler C, Vounatsou P, Smith T: Spatial effects of the social marketing of insecticide-treated nets on malarai morbidity. Trop Med Int Health. 2005, 10: 11-18.View ArticlePubMedGoogle Scholar
- Binka F, Akweongo P: Prevention of malaria using ITNs: Potential for achieving the millenium development goals. Curr Molec Med. 2006, 6 (2): 261-267.View ArticleGoogle Scholar
- Uhlemann AC, Krishna S: Antimalarial multi-drug resistance in Asia: Mechanisms and assessment. Curr Top Microbiol Immunol. 2005, 295: 39-53.Google Scholar
- Coleman M, Hemingway J: Insecticide resistance monitoring and evaluation in disease transmitting mosquitoes. Journal of Pesticide Science. 2007, 32 (2): 69-76.View ArticleGoogle Scholar
- ter Kuile FO, van Eijk AM, Filler SJ: Effect of sulfadoxine-pyrimethamine resistance on the efficacy of intermittent preventive therapy for malaria control during pregnancy - A systematic review. JAMA. 2007, 297 (23): 2603-2616.View ArticlePubMedGoogle Scholar
- Knols BGJ, Louis CE: Bridging Laboratory and Field Research for Genetic Control of Disease Vectors. 2006, Wageningen , Frontis, 210-Google Scholar
- Benedict MQ, Robinson AS: The first releases of transgenic mosquitoes: an argument for the sterile insect technique. Trends Parasitol. 2003, 19 (8): 349-355.View ArticlePubMedGoogle Scholar
- Helinski MEH, El-Sayed B, Knols BGJ: The Sterile Insect Technique: can established technology beat malaria?. Entomologische Berichten (Amsterdam). 2006, 66 (1): 13-20.Google Scholar
- Catteruccia F: Malaria vector control in the third millennium: progress and perspectives of molecular approaches. Pest Manag Sci. 2007, 63: 634-640.View ArticleGoogle Scholar
- Knols BGJ, Bossin HC, Mukabana WR, Robinson AS: Transgenic mosquitoes and the fight against malaria: Managing technology push in a turbulent GMO world. Am J Trop Med Hyg. 2007, 77 (6 Suppl): 232-242.PubMedGoogle Scholar
- Lobo NF, Clayton JR, Fraser MJ, Kafatos FC, Collins FH: High efficiency germ-line transformation of mosquitoes. Nat Protoc. 2006, 1: 1312-1317.View ArticlePubMedGoogle Scholar
- Rodrigues FG, Oliveira SB, Rocha BC, Moreira LA: Germline transformation of Aedes fluviatilis (Diptera : Culicidae) with the piggyBac transposable element. Mem Inst Oswaldo Cruz. 2006, 101 (7): 755-757.View ArticlePubMedGoogle Scholar
- Allen ML, O'Brochta DA, Atkinson PW, Levesque CS: Stable, germ-line transformation of Culex quinquefasciatus (Diptera : Culicidae). J Med Entomol. 2001, 38 (5): 701-710.View ArticleGoogle Scholar
- Catteruccia F, Nolan T, Loukeris TG, Blass C, Savakis C, Kafatos FC, Crisanti A: Stable germline transformation of the malaria mosquito Anopheles stephensi. Nature. 2000, 405 (6789): 959-962.View ArticlePubMedGoogle Scholar
- Moreira LA, Edwards MJ, Adhami F, Jasinskiene N, James AA, Jacobs-Lorena M: Robust gut-specific gene expression in transgenic Aedes aegypti mosquitoes. Proc Natl Acad Sci U S A. 2000, 97 (20): 10895-10898.PubMed CentralView ArticlePubMedGoogle Scholar
- Kim W, Koo H, Richman AM, Seeley D, Vizioli J, Klocko AD, O'Brochta DA: Ectopic expression of a cecropin transgene in the human malaria vector mosquito Anopheles gambiae (Diptera: Culicidae): effects on susceptibility to Plasmodium. J Med Entomol. 2004, 41 (3): 447-455.View ArticlePubMedGoogle Scholar
- Ito J, Ghosh A, Moreira LA, Wimmer EA, Jacobs-Lorena M: Transgenic anopheline mosquitoes impaired in transmission of a malaria parasite. Nature. 2002, 417 (6887): 452-455.View ArticlePubMedGoogle Scholar
- Moreira LA, Ito J, Ghosh A, Devenport M, Zieler H, Abraham EG, Crisanti A, Nolan T, Catteruccia F, Jacobs-Lorena M: Bee venom phospholipase inhibits malaria parasite development in transgenic mosquitoes. J Biol Chem. 2002, 277 (43): 40839-40843.View ArticlePubMedGoogle Scholar
- Franz AWE, Sanchez-Vargas I, Adelman ZN, Blair CD, Beaty BJ, James AA, Olson KE: Engineering RNA interference-based resistance to dengue virus type 2 in genetically modified Aedes aegypti. Proc Natl Acad Sci USA. 2006, 103 (11): 4198-4203.PubMed CentralView ArticlePubMedGoogle Scholar
- Catteruccia F, Godfray HC, Crisanti A: Impact of genetic manipulation on the fitness of Anopheles stephensi mosquitoes. Science. 2003, 299 (5610): 1225-1227.View ArticlePubMedGoogle Scholar
- Irvin N, Hoddle MS, O'Brochta DA, Carey B, Atkinson PW: Assessing fitness costs for transgenic Aedes aegypti expressing the GFP marker and transposase genes. Proc Natl Acad Sci U S A. 2004, 101 (3): 891-896.PubMed CentralView ArticlePubMedGoogle Scholar
- Moreira LA, Wang J, Collins FH, Jacobs-Lorena M: Fitness of Anopheline mosquitoes expressing transgenes that inhibit Plasmodium development. Genetics. 2004, 166: 1337-1341.PubMed CentralView ArticlePubMedGoogle Scholar
- Marrelli MT, Li CY, Rasgon JL, Jacobs-Lorena M: Transgenic malaria-resistant mosquitoes have a fitness advantage when feeding on Plasmodium-infected blood. Proc Natl Acad Sci USA. 2007, 104 (13): 5580-5583.PubMed CentralView ArticlePubMedGoogle Scholar
- Huho BJ, Ng'habi KR, Killeen GF, Nkwengulila G, Knols BGJ, Ferguson HM: Nature beats nurture: a case study of the physiological fitness of free-living and laboratory-reared male Anopheles gambiae s. l. J Exp Biol. 2007, 210 (16): 2939-2947.View ArticlePubMedGoogle Scholar
- Ng'habi KR, Huho BJ, Nkwengulila G, Killeen GF, Knols BGJ, Ferguson HM: Sexual selection in mosquito swarms: may the best man lose?. Animal Behav. 76: 105-112.Google Scholar
- Ng'habi KR, John B, Nkwengulila G, Knols BGM, Killeen GF, Ferguson HM: Effect of larval crowding on mating competitiveness of Anopheles gambiae mosquitoes. Malar J. 2005, 4: 49-PubMed CentralView ArticlePubMedGoogle Scholar
- Ferguson HM, John B, Ng'habi K, Knols BGJ: Redressing the sex imbalance in knowledge of vector biology. Trends Ecol Evol. 2005, 20 (4): 202-209.View ArticlePubMedGoogle Scholar
- Knols BG, Njiru BN, Mathenge EM, Mukabana WR, Beier JC, Killeen GF: MalariaSphere: A greenhouse-enclosed simulation of a natural Anopheles gambiae (Diptera: Culicidae) ecosystem in western Kenya. Malar J. 2002, 1 (1): 19-PubMed CentralView ArticlePubMedGoogle Scholar
- Fansiri T, Thavara U, Tawatsin A, Krasaesub S, Sithiprasasna R: Laboratory and semi-field evaluation of mosquito dunks against Aedes aegypti and Aedies albopictus larvae (Diptera: culicidae). Southeast Asian J Trop Med Public Health. 2006, 37 (1): 62-66.PubMedGoogle Scholar
- Curtis CF: Population replacement in Culex fatigans by means of cytoplasmic incompatibility. 2 Field cage experiments with overlapping generations. Bull W H O. 1976, 53: 107-119.PubMed CentralPubMedGoogle Scholar
- Rowe CL, Dunson WA: The value of simulated pond communities in mesocosms for studies of amphibian ecology and ecotoxicology. J Herpetol. 1994, 28 (3): 346-356.View ArticleGoogle Scholar
- Hackett LW, Bates M: The laboratory for mosquito research in Albania. Trans 3rd Int Cong of Trop Med Malaria. 1939, 2: 113-123.Google Scholar
- Russell PF, Rao TR: On the swarming, mating, and ovipositing behavior of Anopheles culicifacies. Am J Trop Med Hyg. 1942, s1-22: 417-427.Google Scholar
- Curtis CF, Grover KK, Suguna SG, Uppal DK, Dietz K, Agarwal HV, Kazmi SJ: Comparative field cage tests of the population suppressing efficiency of three genetic control systems for Aedes aegypti. Heredity. 1976, 36: 11-29.View ArticlePubMedGoogle Scholar
- Curtis CF, Lorimer N, Rai KS, Suguna SG, Uppal DK, Kazmi SJ, Hallinan E, Dietz K: Simulation of alternative genetic control systems for Aedes aegypt i in outdoor cages and with a computer. J Genetics. 1976, 62: 101-115.View ArticleGoogle Scholar
- Curtis CF, Reuben R: Destruction in the 1970's of a research unit in India on genetic control of mosquitoes and a warning for the future management of transgenic research. Antenna. 2007, 31: 214-216.Google Scholar
- Okanda FM, Dao A, Njiru BN, Arija J, Akelo HA, Toure Y, Odulaja A, Beier JC, Githure JI, Yan G, Gouagna LC, Knols BG, Killeen GF: Behavioural determinants of gene flow in malaria vector populations: Anopheles gambiae males select large females as mates. Malar J. 2002, 1 (1): 10-PubMed CentralView ArticleGoogle Scholar
- Okech BA, Gouagna LC, Walczak E, Kabiru EW, Beier JC, Yan GY, Githure JI: The development of Plasmodium falciparum in experimentally infected Anopheles gambiae (Diptera : Culicidae) under ambient microhabitat temperature in western Kenya. Acta Tropica. 2004, 92 (2): 99-108.View ArticlePubMedGoogle Scholar
- Impoinvil DE, Kongere JO, Foster WA, Njiru BN, Killeen GF, Githure JI, Beier JC, Hassanali A, Knols BGJ: Feeding and survival of the malaria vector Anopheles gambiae Giles on plants growing in Western Kenya. Med Vet Entomol. 2004, 18: 108-115.View ArticlePubMedGoogle Scholar
- Mathenge EM, Killeen GF, O. OD, Irungu LW, Ndegwa PN, Knols BG: Development of an exposure-free bednet trap for sampling Afrotropical malaria vectors. Med Vet Entomol. 2002, 16 (1): 67-74.View ArticlePubMedGoogle Scholar
- Seyoum A, Kabiru EW, Lwande W, Killeen GF, Hassanali A, Knols BGJ: Repellency of live potted plants against Anopheles gambiae from human baits in semi-field experimental huts. Am J Trop Med Hyg. 2002, 67 (2): 191-195.PubMedGoogle Scholar
- Seyoum A, Palsson K, Kung'a S, Kabiru EW, Lwande W, Killeen GF, Hassanali A, Knols BGJ: Traditional use of mosquito-repellent plants in western Kenya and their evaluation in semi-field experimental huts against Anopheles gambiae: ethnobotanical studies and application by thermal expulsion and direct burning. Trans R Soc Trop Med Hyg. 2002, 96 (3): 225-231.View ArticlePubMedGoogle Scholar
- Njiru BN, Mukabana WR, Takken W, Knols BGJ: Trapping of the malaria vector Anopheles gambiae with odour-baited MM-X traps in semi-field conditions in western Kenya. Malar J. 2006, 5: 39-PubMed CentralView ArticlePubMedGoogle Scholar
- Smith T, Charlwood JD, Kihonda J, Mwankusye S, Billingsley P, Meuwissen J, Lyimo E, Takken W, Teuscher T, Tanner M: Absence of seasonal variation in malaria parasitaemia in an area of intense seasonal transmission. Acta Tropica. 1993, 54: 55-72.View ArticlePubMedGoogle Scholar
- Drakeley C, Schellenberg D, Kihonda J, Sousa CA, Arez AP, Lopes D, Lines J, Mshinda H, Lengeler C, Arnmstrong Schellenberg J, Tanner M, Alonso P: An estimation of the entomological inoculation rate for Ifakara: a semi-urban area in a region of intense malaria transmission in Tanzania. Trop Med Int Health. 2003, 8: 767-774.View ArticlePubMedGoogle Scholar
- Kitua AY, Smith T, Alonso PL, Masanja H, Urassa H, Menendez C, Kimario J, Tanner M: Plasmodium falciparum malaria in the first year of life in an area of intense and perennial transmission. Trop Med Int Health. 1996, 1: 475-484.View ArticlePubMedGoogle Scholar
- Charlwood JD, Smith T, Billingsley PF, Takken W, Lyimo EOK, Meuwissen JHET: Survival and infection probabilities of anthropophagic anophelines from an area of high prevalence of Plasmodium falciparum in humans. Bull Ent Res. 1997, 87 (5): 445-453.View ArticleGoogle Scholar
- Smith T, Charlwood JD, Takken W, Tanner M, Spiegelhalter DJ: Mapping densities of malaria vectors within a single village. Acta Tropica. 1995, 58: 1-18.View ArticleGoogle Scholar
- Charlwood JD, Vij R, Billingsley PF: Dry season refugia of malaria-transmitting mosquitoes in a dry savannah zone of east Africa. Am J Trop Med Hyg. 2000, 62 (6): 726-732.PubMedGoogle Scholar
- Benedict MQ, D'Abbs P, Dobson S, Gottlieb M, Harrington LC, Higgs S, James AA, James S, Knols BGJ, Lavery J, O'Neill S, Scott TW, Takken W, Toure Y: Guidance for contained field trials of vector mosquitoes engineered to contain a gene drive system: recommendations of a scientific working group. Vector Borne and Zoonotic Diseases. 8: 127-166.Google Scholar
- Schellenberg D, Menendez C, Aponte J, Guinovart C, Mshinda H, Tanner M, Alonso P: The changing epidemiology of malaria in Ifakara Town, southern Tanzania. Trop Med Int Health. 2004, 9: 68-76.View ArticlePubMedGoogle Scholar
- Charlwood JD, Kihonda J, Sama S, Billingsley PF, Hadji H, Verhave JP, Lyimo EO, Luttikhuizen PC, Smith T: The rise and fall of Anopheles arabiensis (Diptera: Culicidae) in a Tanzanian village. Bull Ent Res. 1995, 85: 37-44.View ArticleGoogle Scholar
- Huho B, Ng'habi K, Killeen GF, Nkwengulila G, Knols BGJ, Ferguson HM: A reliable morphological method to assess the age of male Anopheles gambiae. Malar J. 2006, 5: 26-View ArticleGoogle Scholar
- WHO: Technical consultation on specifications and quality control of netting materials and mosquito nets. 2007, GenevaGoogle Scholar
- Prior A, Torr SJ: Host selection by Anopheles arabiensis and An. quadriannulatus feeding on cattle in Zimbabwe. Med Vet Entomol. 2002, 16 (2): 207-213.View ArticlePubMedGoogle Scholar
- White GB: Anopheles gambiae complex and disease transmission in Africa. Trans R Soc Trop Med Hyg. 1974, 68: 278-301.View ArticlePubMedGoogle Scholar
- Killeen GF, McKenzie FE, Foy BD, Bogh C, Beier JC: The availability of potential hosts as a determinant of feeding behaviours and malaria transmission by African mosquito populations. Trans R Soc Trop Med Hyg. 2001, 95: 469-476.PubMed CentralView ArticlePubMedGoogle Scholar
- Githeko AK, Service MW, Mbogo CM, Atieli F, Juma FO: Origin of blood meals in indoor and outdoor resting malaria vectors in western Kenya. Acta Tropica. 1994, 58: 307-316.View ArticlePubMedGoogle Scholar
- Mwanagangi MM, Mbogo CM, Nzovu JG, Githure JI, Yan G, Beier JC: Blood-meal analysis for anopheline mosquitoes sampled along the Kenyan coast. J Am Mosq Control Assoc. 2003, 19 (4): 371-375.Google Scholar
- Helinski MEH, Hassan MM, El-Motasim WM, Malcolm CA, Knols BGJ, El-Sayed B: Towards a sterile insect technique field release of mosquitoes in Sudan: irradiation, transportation, and field cage experimentation. Malaria J. 7: 65-Google Scholar
- Kirby MJ, Lindsay SW: Responses of adult mosquitoes of two sibling species, Anopheles arabiensis and A. gambiae s.s. (Diptera: Culicidae) to high temperatures. Bull Entomol Res. 2004, 94: 441-448.View ArticlePubMedGoogle Scholar
- Huang J, Walker ED, Vulule J, Miller JR: Daily temperature profiles in and around Western Kenyan larval habitats of Anopheles gambiae as related to egg mortality. Malar J. 2006, 5: 87-PubMed CentralView ArticlePubMedGoogle Scholar
- Charlwood JD, Smith T, Lyimo E, Kitua AY, Masanja H, Booth M, Alonso P, Tanner M: Incidence of Plasmodium falciparum infection in infants in relation to exposure to sporozoite-infected Anophelines. Am J Trop Med Hyg. 1998, 59: 243-251.PubMedGoogle Scholar
- Armstrong JA, Bransby-Williams WR: The maintenance of a colony of Anopheles gambiae with observations on the effects of changes in temperature. Bull W H O. 1961, 24: 427-435.PubMed CentralPubMedGoogle Scholar
- De Meillon B: Entomological studies - observation of Anopheles funestus and Anopheles gambiae in the Transvaal . Publications of the South African Institute for Medical Research. 1934, 6: 195-248.Google Scholar
- Bayoh MN, Lindsay SW: Effect of temperature on the development of the aquatic stages of Anopheles gambiae sensu stricto (Diptera : Culicidae). Bull Entomol Res. 2003, 93 (5): 375-381.View ArticlePubMedGoogle Scholar
- Haddow AJ: Measurements of temperature and light in artificial pools with reference to the larval habitat of Anopheles (Myzomyia) gambiae, Giles, and A. (M.) funestus, Giles. Bull Entomol Res. 1943, 34: 89-93.View ArticleGoogle Scholar
- Huang J, Walker ED, Vulule J, Miller JR: The influence of darkness and visual contrast on oviposition by Anopheles gambiae in moist and dry substrates. Physiology Entomology. 2007, 32: 34-40.View ArticleGoogle Scholar
- Lindsay SW, Parson L, Thomas CJ: Mapping the ranges and relative abundance of the two principal African malaria vectors, Anopheles gambiae sensu stricto and An. arabiensis, using climate data. Proc Biol Sci. 1998, 265: 847-854.PubMed CentralView ArticlePubMedGoogle Scholar
- Killeen GF, McKenzie FE, Foy BD, Schieffelin C, Billingsley PF, Beier JC: A simplified model for predicting malaria entomologic inoculation rates based on entomologic and parasitologic parameters relevant to control. Am J Trop Med Hyg. 2000, 62 (5): 535-544.PubMed CentralPubMedGoogle Scholar
- Lehane MJ: The Biology of Blood-sucking in Insects. 2005, London , Cambridge University Press, 321-2ndView ArticleGoogle Scholar
- Bennett GF: The influence of blood meal type on the fecundity of Aedes (Stegomyia) aegypti L. (Diptera: Culicidae). Can J Zool. 1970, 48 (3): 539-543.View ArticlePubMedGoogle Scholar
- Downe AER, Archer JA: Effects of different blood meal sources on digestion and egg production in Culex tarsalis COQ (Diptera Culicidae). J Med Entomol. 1975, 12: 431-437.View ArticlePubMedGoogle Scholar
- Wilson ML, Litwin TS, Gavin TA, Capkanis MC, Maclean DC, Spielman A: Host-dependent differences in feeding and reproduction of Ixodes dammini (Acari: Ixodidae). J Med Entomol. 1990, 27: 945-954.View ArticlePubMedGoogle Scholar
- Harrington LC, Edman JD, Scott TW: Why do female Aedes aegypti (Diptera: Culicidae) feed preferentially and frequently on human blood?. J Med Entomol. 2001, 38 (3): 411-422.View ArticlePubMedGoogle Scholar
- Emmanuelle-Machado P, Koerich LB, Joukoski D, Carvalho-Pinto C, Grisard ED, Steindel M: Biology of Triatoma klugi Carcavallo, Jurbery, Lent & Galvao 2001 (Heteroptera: Reduviidae) under laboratory conditions: effects of distinct blood sources and susceptibility to Trypansoma cruzi and Trypanosoma rangeli. Mem Inst Oswaldo Cruz. 2002, 97 (4): 585-587.View ArticleGoogle Scholar
- Nieves E, Pimenta PFP: Influence of vertebrate blood meals on the development of Leishmania (Viannia) braziliensis and Leishmania (Leishmania) amazonensis in the sand fly Lutzomyia migonei (Diptera: psychodidae). Am J Trop Med Hyg. 2002, 67 (6): 640-647.PubMedGoogle Scholar
- Gillies MT: Selection for host preference in Anopheles gambiae. Nature. 1964, 203: 852-854.View ArticleGoogle Scholar
- Mukhopadhyay J, Rangel EF, Ghosh K, Munstermann LE: Patterns of genetic variability in colonized strains of Lutzomyia longipalpis (Diptera: Psychodidae) and its consequences. Am J Trop Med Hyg. 1997, 57 (2): 216-221.PubMedGoogle Scholar
- Norris DE, Shurtleff AC, Toure YT, Lanzaro GC: Microsatellite DNA polymorphism and heterozygosity among field and laboratory populations of Anopheles gambiae s. s. (Diptera: Culicidae). J Med Entomol. 2001, 38: 336-340.View ArticlePubMedGoogle Scholar
- Arias L, Bejarano EE, Marquez E, Moncada J, Velez I, Uribe S: Mitochondrial DNA divergence between wild and laboratory populations of Anopheles albimanus Wiedemann (Diptera: Culicidae). Neotropical Entomology. 2005, 34 (3): 499-506.View ArticleGoogle Scholar
- Reisen WK: Lessons from the past: an overview of studies by the University of Maryland and the University of California, Berkeley. Ecological aspects for application of genetically modified mosquitoes. Edited by: Takken W, Scott TW. 2003, Wageningen , Kluwer Academic Press, 25-32.Google Scholar
- Helinski MEH, Hood-Nowotny R, Mayr L, Knols BGJ: Stable isotope-mass spectrometric determination of semen transfer in malaria mosquitoes. J Exp Biol. 2007, 210: 1266-1274.View ArticlePubMedGoogle Scholar
- Hood-Nowotny R, Knols BGJ: Stable isotope methods in biological and ecological studies of arthropods. Entomol Exp Appl. 2007, 124: 3-16.View ArticleGoogle Scholar
- Harrington LC, Ponlawat A, Edman JD, Scott TW, Vermeylen F: Influence of container size, location, and time of day on oviposition patterns of the dengue vector, Aedes aegypti, in Thailand. Vect Borne Zoonotic Dis . 8: 415-423.Google Scholar
This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.