Effect of non-human hosts on the human biting rate of primary and secondary malaria vectors in Tanzania.

BACKGROUND: Malaria vectors vary in feeding preference depending on their innate behaviour, host availability and abundance. Host preference and human biting rate in malaria vectors are key factors in establishing zooprophylaxis and zoopotentiation. This study aimed at assessing the impact of non-human hosts in close proximity to humans on the human biting rate of primary and secondary malaria vectors, with varying host preferences. METHODS: The effect of the presence of non-human hosts in close proximity to the human host on the mean catches per person per night, as a proxy for mosquito biting rate, was measured using mosquito-electrocuting traps (METs), in Sagamaganga, Kilombero Valley, Tanzania. Two experiments were designed: (1) a human versus a calf, each enclosed in a MET, and (2) a human surrounded by three calves versus a human alone, with each human volunteer enclosed individually in a MET spaced 10 m apart. Each experiment was conducted on alternate days and lasted for 36 nights per experiment. During each experiment, the positions of hosts were exchanged daily (except the human in experiment 2). All anopheline mosquitoes caught were assayed for Plasmodium sporozoites using enzyme-linked immunosorbent assay. RESULTS: A total of 20,574 mosquitoes were captured and identified during the study, of which 3608 were anophelines (84.4% primary and 15.6% secondary malaria vectors) and 17,146 were culicines. In experiment 1, the primary malaria vector, Anopheles arabiensis, along with Culex spp. demonstrated a preference for cattle, while the primary vectors, Anopheles funestus, preferred humans. In experiment 2, both primary vectors, An. arabiensis and An. funestus, as well as the secondary vector Anopheles rivolurum, demonstrated behaviours amenable to zooprophylaxis, whereas Culex spp. increased their attraction to humans in the presence of nearby cattle. All anopheline mosquitoes tested negative for sporozoites. CONCLUSIONS: The findings of this study provide support for the zooprophylaxis model for malaria vectors present in the Kilombero Valley, and for the zoopotentiation model, as it pertains to the Culex spp. in the region. However, the factors regulating zooprophylaxis and zoopotentiation are complex, with different species-dependent mechanisms regulating these behaviours, that need to be considered when designing integrated vector management programmes.

Drivers behind co-occurrence patterns between pathogenic bacteria, protozoa, and helminths in populations of the multimammate mouse, Mastomys natalensis.

Advances in experimental and theoretical work increasingly suggest that parasite interactions within a single host can affect the spread and severity of wildlife diseases. Yet empirical data to support predicted co-infection patterns are limited due to the practical challenges of gathering convincing data from animal populations and the stochastic nature of parasite transmission. Here, we investigated co-infection patterns between micro- (bacteria and protozoa) and macroparasites (gastro-intestinal helminths) in natural populations of the multimammate mouse (Mastomys natalensis). Fieldwork was performed in Morogoro (Tanzania), where we trapped 211 M. natalensis and tested their behaviour using a modified open-field arena. All animals were checked for the presence of helminths in their gastro-intestinal tract, three bacteria (Anaplasma, Bartonella, and Borrelia) and two protozoan genera (Babesia and Hepatozoon). Besides the presence of eight different helminth genera (reported earlier), we found that 19% of M. natalensis were positive for Anaplasma, 10% for Bartonella, and 2% for Hepatozoon species. Hierarchical modelling of species communities was used to investigate the effect of the different host-related factors on these parasites' infection probability and community structure. Our results show that the infection probability of Bartonella increased with the host's age, while the infection probability of Anaplasma peaked when individuals reached adulthood. We also observed that less explorative and stress-sensitive individuals had a higher infection probability with Bartonella. Finally, we found limited support for within-host interactions between micro-and macroparasites, as most co-infection patterns could be attributed to host exposure time.

Distance from human dwellings differentially affects the efficacy of a synthetic cattle urine odour lure to trap malaria vectors.

BACKGROUND: Cost-effective outdoor-based devices for surveillance and control of outdoor mosquito vector populations can substantially improve their efficacy when baited with synthetic human and animal odours. This study aimed at assessing the dose-dependent efficacy of a previously developed synthetic cattle urine odour to lure malaria vectors, and other mosquito species, to traps placed at different distances from human dwellings outdoors. METHODS: The efficacy of the cattle urine odour lure was assessed through a 5 × 5 Latin square design, using two sets of 5 Suna traps placed at either 1.5 m or 5 m from an adjacent human dwelling, in the rural village of Sagamaganga, Tanzania. Each trap was deployed with one of four doses of the synthetic cattle urine odour blend or a solvent control (heptane). Traps were rotated daily so that each dose and control visited each position twice over a period of 20 experimental nights. The relative attractiveness of each treatment dose and control was compared using a generalized linear mixed model for each species caught. RESULTS: A total of 1568 mosquitoes were caught, of which 783 were anophelines and 785 were culicines. Of the anophelines, 41.6 and 58.3% were primary and secondary vector species, respectively. Unfed and fed females of the primary vector, Anopheles arabiensis, were caught dose-dependently, close to human dwellings (1.5 m), whereas unfed, fed and gravid secondary vector Anopheles pharoensis females were caught dose-dependently, but at a farther distance from the dwellings (5 m). Females of Culex spp. were caught dose-dependently in similar numbers irrespective of the distance from human dwellings. CONCLUSIONS: This study further clarifies the factors to be considered for the implementation of outdoor trapping using the synthetic cattle urine lure to target exophilic and exophagic malaria vectors, for which efficient surveillance and control tools are currently lacking. The findings resulting from this study make significant progress in providing the needed information to overcome the regulatory obstacles to make this tool available for integrated vector management programs, including registration, as well as evaluation and regulation by the World Health Organization.

Seasonal variation in abundance and blood meal sources of primary and secondary malaria vectors within Kilombero Valley, Southern Tanzania.

BACKGROUND: Existing control tools have significantly reduced malaria over the past two decades. However, progress has been stalled due to increased resistance in primary vectors and the increasing role of secondary vectors. This study aimed to investigate the impact of seasonal change on primary and secondary vector abundance and host preference. Understanding the impact of seasonal dynamics of primary and secondary vectors on disease transmission will inform effective strategies for vector management and control. METHODS: Vector abundance was measured through longitudinal collection of mosquitoes, conducted monthly during the wet and dry seasons, in Sagamaganga, a village in the Kilombero Valley, Tanzania. Mosquitoes were collected indoors using CDC light traps and backpack aspirators, and outdoors using resting buckets baited with cattle urine. In addition, a direct measure of host preference was taken monthly using human- and cattle-baited mosquito electrocuting traps. A host census was conducted to provide an indirect measure of host preference together with monthly blood meal source analysis. All collected mosquitoes were assayed for Plasmodium sporozoites. RESULTS: A total of 2828 anophelines were collected, of which 78.5% and 21.4%, were primary and secondary vectors, respectively. The abundance of the primary vectors, Anopheles arabiensis and Anopheles funestus, and of the secondary vectors varied seasonally. Indirect measures of host preference indicated that all vectors varied blood meal choice seasonally, with the direct measure confirming this for An. arabiensis. All anopheline mosquitoes tested negative for sporozoites. CONCLUSIONS: At the study location, the abundance of both primary and secondary vectors changed seasonally. Indirect and direct measures of host preference demonstrated that An. arabiensis varied from being zoophilic to being more opportunistic during the wet and dry seasons. A similar trend was observed for the other vectors.

Retrospective Analysis of Malaria Cases in a Potentially High Endemic Area of Morogoro Rural District, Eastern Tanzania.

BACKGROUND: Malaria is increasingly characterized by appreciable fine-scale variability in ecology and topography, and it is likely that we are missing some salient foci with unprecedented malaria transmission intensity in different parts of Tanzania. Therefore, efforts aimed at identifying area-specific malaria situation and intervening are needed to preserve the realized health gains and achieve elimination. Mkuyuni and Kiroka, adjacent wards within Morogoro Rural District, are purported to form one of such foci. PATIENTS AND METHODS: A retrospective study was conducted to determine six-year (2014-2019) malaria prevalence rates based on outpatients and laboratory registers obtained from two health facilities, one per ward, carrying out diagnosis of malaria either through microscopy or malaria rapid diagnostic test (mRDT). These data were checked for completeness before carrying out statistical analysis. RESULTS: Overall, 35,386 (46.19%) out of 76,604 patients were positive for malaria. The average proportion of malaria cases was significantly higher in Mkuyuni (51.23%; n=19,438) than Kiroka (41.21%; n = 15,938) (P <0.001). Females were more affected than males (P <0.001);, and irrespective of the sex, most malaria cases were recorded in children <5 years of age (P <0.001) except at Mkuyuni. Malaria was recorded virtually all year round; however, the highest proportion of cases was recorded in April and July (P <0.001). CONCLUSION: This study revealed high malaria endemicity in Mkuyuni and Kiroka, with prevalence rate as high as 60.98%, which is far higher than the overall national average prevalence of 9%. More studies are needed in these and other putatively high endemic foci in Tanzania in order to inform the future course of action in disease surveillance and control.

Passive Outdoor Host Seeking Device (POHD): Designing and Evaluation against Outdoor Biting Malaria Vectors.

Odor-baited devices are increasingly needed to compliment long-lasting insecticidal nets (LLINs) and indoor residual spraying (IRS) for control of residual malaria transmission. However, the odor-baited devices developed so far are bulky, dependent on the source of electricity and carbon dioxide (CO2), and they are logistically unsuitable for scaling up in surveillance and control of malaria vectors. We designed a passive and portable outdoor host seeking device (POHD) and preliminarily evaluated suitable components against Anopheles arabiensis that maintains residual malaria transmission. Experiments were conducted using semifield reared An. arabiensis within the semifield system at Ifakara Health Institute (IHI) in southeastern Tanzania. These mosquitoes were exposed to Suna traps® baited with BG lures or source of light and augmented with carbon dioxide (CO2) in view of identifying best attractants necessary to improve attractiveness of designed POHD. Two Suna traps® were hanged at the corner but outside the experimental hut in a diagonal line and rotated between four corners to control for the effect of position and wind direction on mosquito catches. Furthermore, mosquitoes were also exposed to either a bendiocarb-treated or bendiocarb-untreated POHD baited with Mbita blend, Ifakara blend, and worn socks and augmented with warmth (i.e., 1.5 liter bottle of warm water) inside an experimental hut or a screened rectangular box. This study demonstrated that mosquitoes were more strongly attracted to Suna trap® baited with BG lures and CO2 relative to those traps baited with a source of light and CO2. The POHD baited with synthetic blends attracted and killed greater proportion of An. arabiensis compared with POHD baited with worn socks. Efficacy of the POHD was unaffected by source of warmth, and it was reduced by about 50% when the device was tested inside a screened rectangular box relative to closed experimental hut. Overall, this study demonstrates that the POHD baited with synthetic blends (Mbita and Ifakara blends) and bendiocarb can effectively attract and kill outdoor biting malaria vector species. Such POHD baited with synthetic blends may require the source of CO2 to enhance attractiveness to mosquitoes. Further trials are, therefore, ongoing to evaluate attractiveness of improved design of POHD baited with slow-release formulation of synthetic blends and sustainable source of CO2 to malaria vectors under semifield and natural environments.

Semifield Evaluation of Improved Passive Outdoor Host Seeking Device (POHD) for Outdoor Control of Anopheles arabiensis Mosquitoes.

Despite the considerable progress made so far, the effectiveness and mass application of odour-baited outdoor mosquito control devices in pipelines is limited by several factors. These include the design and size of the devices, optimal placement of attractive blends, and nature of materials into which the blends are impregnated. The primary aim of this study was to manipulate these factors to improve the attractiveness of our recently developed passive outdoor host seeking device (POHD) to outdoor biting Anopheles arabiensis. Specifically, the study aimed to determine optimal placement of odour blends and killing bioactives in POHD for maximum attraction and killing of An. arabiensis and to assess the effects of blend types, formulation, and residual activity on attractiveness of the POHD to An. arabiensis. The POHDs baited with attractive blends, carbon dioxide (CO2), and bendiocarb-treated electrostatic netting were placed either towards the top or bottom openings, and other modifications were exposed to An. arabiensis under the semifield system at Ifakara Health Institute (IHI). Each night, a total of 100 starved female, 3-7-day-old, semifield reared An. arabiensis mosquitoes were released, collected the next morning (alive or dead), counted, and recorded. Live mosquitoes were maintained in the semifield insectary and monitored for 24 hours mortality. Each treatment combination of the POHD was tested in three replicates. Overall, the results indicated that the proportion of mosquitoes attracted to and killed in the POHD varied with position of attractants and killing agent (bendiocarb). The POHD with bottom placed attractants and bendiocarb attracted and killed higher proportion of mosquitoes compared to the POHD with top placed attractants and bendiocarb. The highest mortalities were observed when the POHD was baited with a combination of attractive blends and CO2. Moreover, the residual activity of attractive blends applied inside POHD varied with type and formulation of attractive blend. The POHD packed with Mbita and Ifakara blend in microencapsulated pellets (granules) attracted higher proportion of mosquitoes than that baited with soaked nylon-strip formulation of either blends. Interestingly, POHD baited with Mbita blend in microencapsulated pellets (granules) formulation attracted and killed higher proportion of mosquitoes (>90%) than that baited with Ifakara blend even 9 months after application. Conclusively, the POHD remained effective for a relatively longer period of time when baited with bottom placed synthetic blends and CO2 combination, thus warranting further trials under real life situations.

The Use of Granular Cyclopentanone as Alternative to Artificial Source of Carbon Dioxide in Improved Passive Outdoor Host Seeking Device (POHD).

Reliable sources of CO2 that are relatively cheap, obtainable, and easy to sustain are immediately required for scaling up of odor-baited mosquito surveillance and control devices. Several odor-baited devices are in the pipeline; however, their scale-up against residual malaria transmission, particularly in resource poor areas, is limited by the unavailability of reliable sources of CO2 and reliance on electrical power sources among other factors. We evaluated the use of granular cyclopentanone as an alternative to artificial or yeast fermentation-produced CO2 in passive outdoor host seeking device (POHD). Experiments were conducted against semifield reared An. arabiensis within the semifield system (SFS) at Ifakara Health Institute. Mosquitoes were tested against odor-baited POHDs augmented with yeast fermentation-produced CO2, granular cyclopentanone, attractive blends (Mbita or Ifakara), or their combinations. An insecticide, bendiocarb, was a killing agent used as a proxy for marking the mosquitoes visit the POHDs. Relative attractiveness of different treatment combinations was compared based on the proportion of dead mosquitoes that visited the POHD. The POHD augmented with granules of cyclopentanone alone was attractive to An. arabiensis as much as, or more than, POHDs augmented with yeast fermentation-produced CO2. The POHD baited with CO2 attracted more mosquitoes than those POHDs baited with synthetic blends alone; when these blends are combined with CO2, they attracted more mosquitoes than individual blends. More importantly, such POHDs baited with cyclopentanone attracted far greater proportion of mosquitoes than the POHD baited with either Mbita or Ifakara blend alone. The granular cyclopentanone strongly enhanced/potentiated the attractiveness of POHD baited with Mbita blends against mosquitoes compared to that of POHD baited with Ifakara blend. Moreover, the granular cyclopentanone retained its residual activity against An. arabiensis for up to 2 months after application particularly when used in combination with Mbita blend. In conclusion, this study demonstrates that cyclopentanone granules have the potential to substitute sources of CO2 in outdoor-based surveillance and control devices, thus warranting evaluation of such alternative under realistic field conditions.

Community perceptions on outdoor malaria transmission in Kilombero Valley, Southern Tanzania.

BACKGROUND: The extensive use of indoor residual spraying (IRS) and insecticide-treated nets (ITNs) in Africa has contributed to a significant reduction in malaria transmission. Even so, residual malaria transmission persists in many regions, partly driven by mosquitoes that bite people outdoors. In areas where Anopheles gambiae s.s. is a dominant vector, most interventions target the reduction of indoor transmission. The increased use of ITNs/LLINs and IRS has led to the decline of this species. As a result, less dominant vectors such as Anopheles funestus and Anopheles arabiensis, both also originally indoor vectors but are increasingly biting outdoors, contribute more to residual malaria transmission. The study reports the investigated community perceptions on malaria and their implications of this for ongoing outdoor malaria transmission and malaria control efforts. METHODS: This was a qualitative study conducted in two rural villages and two peri-urban areas located in Kilombero Valley in south-eastern Tanzania. 40 semi-structured in-depth interviews and 8 focus group discussions were conducted with men and women who had children under the age of five. The Interviews and discussions focused on (1) community knowledge of malaria transmission, and (2) the role of such knowledge on outdoor malaria transmission as a contributing factor to residual malaria transmission. RESULTS: The use of bed nets for malaria prevention has been stressed in a number of campaigns and malaria prevention programmes. Most people interviewed believe that there is outdoor malaria transmission since they use interventions while indoors, but they are unaware of changing mosquito host-seeking behaviour. Participants pointed out that they were frequently bitten by mosquitoes during the evening when outdoors, compared to when they were indoors. Most participants stay outdoors in the early evening to undertake domestic tasks that cannot be conducted indoors. House structure, poor ventilation and warm weather conditions were reported to be the main reasons for staying outdoors during the evening. Participants reported wearing long sleeved clothes, fanning and slapping themselves, using repellents, and burning cow dung and neem tree leaves to chase away mosquitoes. CONCLUSIONS: Community understanding of multiple prevention strategies is crucial given changes in mosquito host seeking behaviour and the increased incidence of outdoor biting. The current low use of outdoor control measures is attributed largely to limited awareness of outdoor transmission. Improved community understanding of outdoor malaria transmission is critical: efforts to reduce or eliminate malaria transmission will not be successful if the control of outdoor transmission is not emphasized.

Ivermectin-treated cattle reduces blood digestion, egg production and survival of a free-living population of Anopheles arabiensis under semi-field condition in south-eastern Tanzania.

BACKGROUND: Anopheles arabiensis feed on cattle and contributes to residual transmission of malaria in areas with high coverage of long-lasting insecticide-treated nets and indoor residual spraying in East Africa. This study aimed to evaluate the effects of ivermectin-treated cattle as a complementary vector control tool against population of An. arabiensis under the semi-field conditions in south-eastern Tanzania. METHODS: The free-living population of An. arabiensis was allowed to forage on untreated or ivermectin-treated cattle in alternating nights within the semi-field system in south-eastern Tanzania. Fresh blood fed mosquitoes were collected in the morning using mouth aspirators and assessed for their blood meal digestion, egg production, and survivorship. The residual activity of ivermectin-treated cattle was also determined by exposing mosquitoes to the same treatments after every 2 days until day 21 post-treatments. These experiments were replicated 3 times using different individual cattle. RESULTS: Overall, the ivermectin-treated cattle reduced blood meal digestion in the stomach of An. arabiensis, and their subsequent egg production and survival over time. The ivermectin-treated cattle halved blood meal digestion in mosquitoes, but reduced their egg production for up to 15 days. The ivermectin-treated cattle reduced the survival, and median survival times (1-3 days) of An. arabiensis than control cattle. The daily mortality rates of mosquitoes fed on ivermectin-treated cattle increased by five-fold relative to controls in the first week, and it gradually declined up to 21 days after treatment. CONCLUSION: This study demonstrates that long-lasting effects of ivermectin-treated cattle on egg production and survival of An. arabiensis may sustainably suppress their vector density, and reduce residual transmission of malaria. This study suggests that ivermectin-treated non-lactating cattle (i.e. calves, heifers and bulls) could be suitable option for large-scale malaria vector control without limiting consumption of milk and meat by communities in rural settings. Furthermore, simulation models are underway to predict the impact of ivermectin-treated cattle alone, or in combination with LLIN/IRS, the frequency of treatment, and their coverage required to significantly suppress population of An. arabiensis and reduce residual transmission of malaria.

Eave tubes for malaria control in Africa: initial development and semi-field evaluations in Tanzania.

BACKGROUND: Presented here are a series of preliminary experiments evaluating "eave tubes"-a technology that combines house screening with a novel method of delivering insecticides for control of malaria mosquitoes. METHODS: Eave tubes were first evaluated with overnight release and recapture of mosquitoes in a screened compartment containing a hut and human sleeper. Recapture numbers were used as a proxy for overnight survival. These trials tested physical characteristics of the eave tubes (height, diameter, angle), and different active ingredients (bendiocarb, LLIN material, fungus). Eave tubes in a hut with closed eaves were also compared to an LLIN protecting a sleeper in a hut with open eaves. Eave tubes were then evaluated in a larger compartment containing a self-replicating mosquito population, vegetation, and multiple houses and cattle sheds. In this "model village", LLINs were introduced first, followed by eave tubes and associated house modifications. RESULTS: Initial testing suggested that tubes placed horizontally and at eave height had the biggest impact on mosquito recapture relative to respective controls. Comparison of active ingredients suggested roughly equivalent effects from bendiocarb, LLIN material, and fungal spores (although speed of kill was slower for fungus). The impact of treated netting on recapture rates ranged from 50 to 70 % reduction relative to controls. In subsequent experiments comparing bendiocarb-treated netting in eave tubes against a standard LLIN, the effect size was smaller but the eave tubes with closed eaves performed at least as well as the LLIN with open eaves. In the model village, introducing LLINs led to an approximate 60 % reduction in larval densities and 85 % reduction in indoor catches of host-seeking mosquitoes relative to pre-intervention values. Installing eave tubes and screening further reduced larval density (93 % relative to pre intervention values) and virtually eliminated indoor host-seeking mosquitoes. When the eave tubes and screening were removed, larval and adult catches recovered to pre-eave tube levels. CONCLUSIONS: These trials suggest that the "eave tube" package can impact overnight survival of host-seeking mosquitoes and can suppress mosquito populations, even in a complex environment. Further testing is now required to evaluate the robustness of these findings and demonstrate impact under field conditions.

Eave tubes for malaria control in Africa: an introduction.

In spite of massive progress in the control of African malaria since the turn of the century, there is a clear and recognized need for additional tools beyond long-lasting insecticide-treated bed nets (LLINs) and indoor residual spraying (IRS) of insecticides, to progress towards elimination. Moreover, widespread and intensifying insecticide resistance requires alternative control agents and delivery systems to enable development of effective insecticide resistance management strategies. This series of articles presents a novel concept for malaria vector control, the 'eave tube', which may fulfil these important criteria. From its conceptualization to laboratory and semi-field testing, to demonstration of potential for implementation, the stepwise development of this new vector control approach is described. These studies suggest eave tubes (which comprise a novel way of delivering insecticides plus screening to make the house more 'mosquito proof') could be a viable, cost-effective, and acceptable control tool for endophilic and endophagic anophelines, and possibly other (nuisance) mosquitoes. The approach could be applicable in a wide variety of housing in sub-Saharan Africa, and possibly beyond, for vectors that use the eave as their primary house entry point. The results presented in these articles were generated during an EU-FP7 funded project, the mosquito contamination device (MCD) project, which ran between 2012 and 2015. This was a collaborative project undertaken by vector biologists, product developers, modellers, materials scientists, and entrepreneurs from five different countries.

Electrostatic coating enhances bioavailability of insecticides and breaks pyrethroid resistance in mosquitoes.

Insecticide resistance poses a significant and increasing threat to the control of malaria and other mosquito-borne diseases. We present a novel method of insecticide application based on netting treated with an electrostatic coating that binds insecticidal particles through polarity. Electrostatic netting can hold small amounts of insecticides effectively and results in enhanced bioavailability upon contact by the insect. Six pyrethroid-resistant Anopheles mosquito strains from across Africa were exposed to similar concentrations of deltamethrin on electrostatic netting or a standard long-lasting deltamethrin-coated bednet (PermaNet 2.0). Standard WHO exposure bioassays showed that electrostatic netting induced significantly higher mortality rates than the PermaNet, thereby effectively breaking mosquito resistance. Electrostatic netting also induced high mortality in resistant mosquito strains when a 15-fold lower dose of deltamethrin was applied and when the exposure time was reduced to only 5 s. Because different types of particles adhere to electrostatic netting, it is also possible to apply nonpyrethroid insecticides. Three insecticide classes were effective against strains of Aedes and Culex mosquitoes, demonstrating that electrostatic netting can be used to deploy a wide range of active insecticides against all major groups of disease-transmitting mosquitoes. Promising applications include the use of electrostatic coating on walls or eave curtains and in trapping/contamination devices. We conclude that application of electrostatically adhered particles boosts the efficacy of WHO-recommended insecticides even against resistant mosquitoes. This innovative technique has potential to support the use of unconventional insecticide classes or combinations thereof, potentially offering a significant step forward in managing insecticide resistance in vector-control operations.

Epilepsy and tropical parasitic infections in Sub-Saharan Africa: A review.

Several reports have suggested that the high prevalence of epilepsy in sub-Saharan Africa is associated with the high prevalence of parasitic infections affecting the central nervous system. Though epidemiological evidence suggests an association between parasitic infections and epilepsy, the biological causal relationship has not been fully demonstrated for many of these infections. The objective of this paper is to review the available epidemiological evidence on the links between parasitic infections and epilepsy, the pathogenesis and the current gap of knowledge indicating the areas requiring further research. Data for this review were identified and collected using manual and electronic search strategies of published and unpublished sources. In Sub-Saharan Africa, the epidemiology of epilepsy remains unclear and given the differing study designs, the results of available epidemiological studies are difficult to interpret and compare. Evidence from surveys reported a median prevalence of 1.5%. Co-infection of parasitic infections and epilepsy in sub-Saharan Africa are common, particularly in areas characterized by poor hygiene standards. There is an epidemiological link on the association between epilepsy and various parasitic infections. However, the biological causal relationship requires further investigation in adequately designed studies. In conclusion, although several epidemiological and case control studies indicate a relationship between parasitic agents and epilepsy in sub-Saharan Africa, there is a considerable gap of knowledge on the cause and magnitude of the association. Thus, there is an urgent need for systematic epidemiological studies to understand the burden of epilepsy in areas endemic due to preventable parasitic infections, to prove a causal relationship, and to understand the impact of controlling these parasitic diseases on reduction of the burden of epilepsy.

Entomopathogenic fungi, Metarhizium anisopliae and Beauveria bassiana reduce the survival of Xenopsylla brasiliensis larvae (Siphonaptera: Pulicidae).

BACKGROUND: Entomopathogenic fungi, particularly those belonging to the genera Metarhizium and Beauveria have shown great promise as arthropod vector control tools. These agents, however, have not been evaluated against flea vectors of plague. FINDINGS: A 3-h exposure to the fungi coated paper at a concentration of 2 × 108 conidia m-2 infected >90% of flea larvae cadavers in the treatment groups. The infection reduced the survival of larvae that had been exposed to fungus relative to controls. The daily risk of dying was four- and over three-fold greater in larvae exposed to M. anisopliae (HR = 4, p<0.001) and B. bassiana (HR = 3.5, p<0.001) respectively. Both fungi can successfully infect and kill larvae of X. brasiliensis with a pooled median survival time (MST±SE) of 2 ± 0.31 days post-exposure. CONCLUSION: These findings justify further research to investigate the bio-control potential of entomopathogenic fungi against fleas.

Does Cattle Milieu Provide a Potential Point to Target Wild Exophilic Anopheles arabiensis (Diptera: Culicidae) with Entomopathogenic Fungus? A Bioinsecticide Zooprophylaxis Strategy for Vector Control.

Background. Anopheles arabiensis is increasingly dominating malaria transmission in Africa. The exophagy in mosquitoes threatens the effectiveness of indoor vector control strategies. This study aimed to evaluate the effectiveness of fungus against An. arabiensis when applied on cattle and their environments. Methods. Experiments were conducted under semi-field and small-scale field conditions within Kilombero valley. The semi-field reared females of 5-7 days old An. arabiensis were exposed to fungus-treated and untreated calf. Further, wild An. arabiensis were exposed to fungus-treated calves, mud-huts, and their controls. Mosquitoes were recaptured the next morning and proportion fed, infected, and survived were evaluated. Experiments were replicated three times using different individuals of calves. Results. A high proportion of An. arabiensis was fed on calves (>0.90) and become infected (0.94) while resting on fungus-treated mud walls than on other surfaces. However, fungus treatments reduced fecundity and survival of mosquitoes. Conclusion. This study demonstrates for the first time the potential of cattle and their milieu for controlling An. arabiensis. Most of An. arabiensis were fed and infected while resting on fungus-treated mud walls than on other surfaces. Fungus treatments reduced fecundity and survival of mosquitoes. These results suggest deployment of bioinsecticide zooprophylaxis against exophilic An. arabiensis.

Exploiting the behaviour of wild malaria vectors to achieve high infection with fungal biocontrol agents.

BACKGROUND: Control of mosquitoes that transmit malaria has been the mainstay in the fight against the disease, but alternative methods are required in view of emerging insecticide resistance. Entomopathogenic fungi are candidate alternatives, but to date, few trials have translated the use of these agents to field-based evaluations of their actual impact on mosquito survival and malaria risk. Mineral oil-formulations of the entomopathogenic fungi Metarhizium anisopliae and Beauveria bassiana were applied using five different techniques that each exploited the behaviour of malaria mosquitoes when entering, host-seeking or resting in experimental huts in a malaria endemic area of rural Tanzania. RESULTS: Survival of mosquitoes was reduced by 39-57% relative to controls after forcing upward house-entry of mosquitoes through fungus treated baffles attached to the eaves or after application of fungus-treated surfaces around an occupied bed net (bed net strip design). Moreover, 68 to 76% of the treatment mosquitoes showed fungal growth and thus had sufficient contact with fungus treated surfaces. A population dynamic model of malaria-mosquito interactions shows that these infection rates reduce malaria transmission by 75-80% due to the effect of fungal infection on adult mortality alone. The model also demonstrated that even if a high proportion of the mosquitoes exhibits outdoor biting behaviour, malaria transmission was still significantly reduced. CONCLUSIONS: Entomopathogenic fungi strongly affect mosquito survival and have a high predicted impact on malaria transmission. These entomopathogens represent a viable alternative for malaria control, especially if they are used as part of an integrated vector management strategy.

Survival of anopheline eggs and their susceptibility to infection with Metarhizium anisopliae and Beauveria bassiana under laboratory conditions.

The viability of Anopheles gambiae sensu stricto and Anopheles arabiensis (Diptera: Culicidae) eggs over time and the ovicidal activity of Beauveria bassiana (Ascomycota: Cordycipitaceae) and Metarhizium anisopliae (Ascomycota: Clavicipitaceae) were investigated. Eggs were incubated in soil or leaf litter for up to 12 weeks at 26°C and 75%, 86% or >98% relative humidity (RH). Eggs were treated topically with M. anisopliae ICIPE-30 or B. bassiana I93-825 conidia in either water or oil-in-water formulations. Survival of eggs whether treated or not with fungus was similar, and untreated eggs generally did not survive longer than 2 weeks regardless of the substrate or humidity tested. After a minimal 5-day exposure, M. anisopliae at 5 × 10(6) conidia/cm(2) clearly reduced the number of larvae. The efficacy of the fungus increased when it was oil-in-water formulated, and eclosion was completely prevented regardless of the conidial concentration (10(5)-10(7) conidia/cm(2)) after a 10-day exposure in soils at >98% RH. Treatment of eggs with B. bassiana, however, failed to reduce the number of eclosing larvae. This is the first demonstration of the ovicidal activity by M. anisopliae against either A. gambiae s. s. or A. arabiensis and the results underline the potential of this fungus against anopheline mosquitoes.

Infection of Anopheles gambiae mosquitoes with entomopathogenic fungi: effect of host age and blood-feeding status.

Physiological characteristics of insects can influence their susceptibility to fungal infection of which age and nutritional status are among the most important. An understanding of host-pathogen interaction with respect to these physiological characteristics of the host is essential if we are to develop fungal formulations capable of reducing malaria transmission under field conditions. Here, two independent bioassays were conducted to study the effect of age and blood-feeding status on fungal infection and survival of Anopheles gambiae s.s. Giles. Mosquitoes were exposed to 2 × 10(10) conidia m(-2) of oil-formulated Metarhizium anisopliae ICIPE-30 and of Beauveria bassiana I93-825, respectively, and their survival was monitored daily. Three age groups of mosquitoes were exposed, 2-4, 5-8, and 9-12 days since emergence. Five groups of different feeding status were exposed: non-blood-fed, 3, 12, 36, and 72 h post-blood feeding. Fungal infection reduced the survival of mosquitoes regardless of their age and blood-feeding status. Although older mosquitoes died relatively earlier than younger ones, age did not tend to affect mosquito susceptibility to fungal infection. Non-blood-fed mosquitoes were more susceptible to fungus infection compared to all categories of blood-fed mosquitoes, except for those exposed to B. bassiana 72 h post-blood feeding. In conclusion, formulations of M. anisopliae and B. bassiana can equally affect mosquitoes of different age classes, with them being relatively more susceptible to fungus infection when non-blood-fed.

Severe tungiasis in Western Tanzania: case series.

Tungiasis is caused by infestation with the sand flea (Tunga penetrans). This ectoparasitosis is endemic in economically depressed communities in South American and African countries. However, data on the epidemiology of tungiasis in Tanzania are very limited and the disease does not receive much attention from health care professionals. During a community cross sectional survey in northwest Tanzania, we identified five individuals extremely infested with high number of parasites. A total of 435 lesions were recorded with patients presenting with >75 lesions and showed signs of intense acute and chronic inflammation. Superinfection of the lesions characterized by pustule formation, suppuration and ulceration were common. Loss of nails and walking difficulty was also observed. In Tanzanian communities living under extreme poverty characterized by poor housing condition and inadequate health services, tungiasis may cause severe morbidities. Further studies on risk factors and disease-related behavior of affected populations are needed to design adequate control measures.