Laboratory and semi-field evaluation of the efficacy of Bacillus thuringiensis var. israelensis (Bactivec®) and Bacillus sphaericus (Griselesf®) for control of mosquito vectors in northeastern Tanzania.

Bacterial larvicides Bacillus thuringiensis var. israelensis (Bti) and Bacillus sphaericus (Bs) have been used extensively for mosquito control. However, their efficacy varies greatly mainly due to factors related to target mosquitoes, larval habitat conditions, and inherent larvicide properties. We evaluated the efficacy of Bti (Bactivec®) and Bs (Griselesf®) for control of Anopheles gambiae complex, Culex quinquefasciatus and Aedes aegypti larvae under laboratory and semi-field conditions in northeastern Tanzania. Laboratory bioassays were conducted with five to six different concentrations of Bti and Bs, replicated four times and the experiment repeated on three different days. Larvae mortality was recorded at 24 or 48 h after the application of larvicide and subjected to Probit analysis. Laboratory bioassays were followed by semi-field trials to establish initial and residual activity of Bti and Bs. Semi-field trials were conducted in artificial larval habitats in the open sunlit ground and in "mosquito spheres". These artificial larval habitats were colonized with mosquito larvae, treated with Bti and Bs, and the impact of treatments on mosquito larvae was monitored daily. Lethal concentration values that caused 50% and 95% mortalities of test larvae (LC50 and LC95) showed that An. gambiae complex and Cx. quinquefasciatus tested were highly susceptible to Bti and Bs under laboratory conditions. Likewise, larvae of Ae. aegypti were highly susceptible to Bti, with LC95 value as low as 0.052 mg/l. However, Ae. aegypti larvae were not susceptible to Bs under practical doses of laboratory settings. In semi-field trials, all treatment dosages for Bti provided 91.0-100% larval mortality within 24 h whereas Bs resulted in 96.8-100% larval mortality within the same time-frame. Bs had a more prolonged residual activity, with pupal reductions range of 55.7-100% for 9 days at all application rates while the corresponding pupal reduction with Bti was 15.4-100% for 5 days. Due to the low residual activity of Bti and Bs tested, weekly application at a maximum label rate would be appropriate to reduce mosquito larvae in natural larval habitats. Based on laboratory findings, Bs product tested would not be recommended for use in the control of Ae. aegypti.

The effectiveness of non-pyrethroid insecticide-treated durable wall lining to control malaria in rural Tanzania: study protocol for a two-armed cluster randomized trial.

BACKGROUND: Despite considerable reductions in malaria achieved by scaling-up long-lasting insecticidal nets (LLINs) and indoor residual spraying (IRS), maintaining sustained community protection remains operationally challenging. Increasing insecticide resistance also threatens to jeopardize the future of both strategies. Non-pyrethroid insecticide-treated wall lining (ITWL) may represent an alternate or complementary control method and a potential tool to manage insecticide resistance. To date no study has demonstrated whether ITWL can reduce malaria transmission nor provide additional protection beyond the current best practice of universal coverage (UC) of LLINs and prompt case management. METHODS/DESIGN: A two-arm cluster randomized controlled trial will be conducted in rural Tanzania to assess whether non-pyrethroid ITWL and UC of LLINs provide added protection against malaria infection in children, compared to UC of LLINs alone. Stratified randomization based on malaria prevalence will be used to select 22 village clusters per arm. All 44 clusters will receive LLINs and half will also have ITWL installed on interior house walls. Study children, aged 6 months to 11 years old, will be enrolled from each cluster and followed monthly to estimate cumulative incidence of malaria parasitaemia (primary endpoint), time to first malaria episode and prevalence of anaemia before and after intervention. Entomological inoculation rate will be estimated using indoor CDC light traps and outdoor tent traps followed by detection of Anopheles gambiae species, sporozoite infection, insecticide resistance and blood meal source. ITWL bioefficacy and durability will be monitored using WHO cone bioassays and household surveys, respectively. Social and cultural factors influencing community and household ITWL acceptability will be explored through focus-group discussions and in-depth interviews. Cost-effectiveness, compared between study arms, will be estimated per malaria case averted. DISCUSSION: This protocol describes the large-scale evaluation of a novel vector control product, designed to overcome some of the known limitations of existing methods. If ITWL is proven to be effective and durable under field conditions, it may warrant consideration for programmatic implementation, particularly in areas with long transmission seasons and where pyrethroid-resistant vectors predominate. Trial findings will provide crucial information for policy makers in Tanzania and other malaria-endemic countries to guide resource allocations for future control efforts. TRIAL REGISTRATION: NCT02533336 registered on 13 July 2014.

Spatial abundance and human biting rate of Anopheles arabiensis and Anopheles funestus in savannah and rice agro-ecosystems of Central Tanzania.

This study was carried out to determine the spatial variations in malaria mosquito abundance and human biting rate in five villages representing rice-irrigation and savannah ecosystems in Kilosa District, central Tanzania. The study involved five villages namely Tindiga and Malui (wetland/rice irrigation), Twatwatwa and Mbwade (dry savannah) and Kimamba (wet savannah). Indoor mosquitoes were sampled using Centers for Disease Control and Prevention light traps in three houses in each village. Anopheles gambiae s.l. molecular identification was carried out using polymerase chain reaction (PCR). A total of 936 female mosquitoes were collected. About half (46.9%) were malaria mosquitoes (Anopheles gambiae s.l.=28.6%; An. funestus= 18.3%). A total of 161 (60.1%) of the morphologically identified An. gambiae s.l. (268) and subjected to PCR analysis for speciation were genotyped as An. arabiensis. The An. funestus complex mosquitoes were composed of An. funestus funestus and An. rivulorum at the 5:1 ratio. On average, 17.9 Anopheles mosquitoes were collected per village per day. Two-thirds (62.8%) of the malaria mosquitoes were collected in Malui (rice agro-ecosystem) and the lowest number (2.3%) in Twatwatwa (dry savannah ecosystem). The biting rate per person per night for An. arabiensis+An. funestus s.s. was highest in Malui (46.0) and lowest in Twatwatwa (1.67). The parity rate of the An. funestus mosquitoes was lower compared to that of An. arabiensis and none of the mosquitoes was infected with malaria sporozoites. In conclusion, An. arabiensis is the most abundant malaria vector in Kilosa district and its variation is related to the ecological system. The heterogeneity in malaria mosquito abundance and human biting rate could be used to guide selection of locally appropriated control interventions.

Malaria, anaemia and nutritional status among schoolchildren in relation to ecosystems, livelihoods and health systems in Kilosa District in central Tanzania.

BACKGROUND: Malaria prevalence and transmission intensity in Tanzania is heterogeneous with spatial and temporal variations between geographical areas and ecological systems. The objective of this study was to determine the prevalence of malaria, anaemia and nutritional status in relation to livelihoods, ecosystem and health systems in Kilosa District in central Tanzania. METHODS: This study was conducted in four villages, two characterised by rice irrigation ecosystem and the other two by dry savannah ecosystem and pastoral livelihoods. In each ecosystem, one of the villages had a healthcare facility. Schoolchildren were screened for malaria infection using malaria rapid diagnostic test (mRDT) and microscopy and they were assessed for their anaemia and nutritional statuses. RESULTS: A total of 1,019 school children (age = 4-16 years) were screened for malaria infection. The overall prevalence of Plasmodium falciparum infection was 10.6% and 4.5% by mRDT and microscopy, respectively. Children from pastoral villages had lower (2.9%) prevalence of malaria than their counterparts (18.2%) in the rice irrigation villages. A significantly high risk of malaria was observed among children in rice irrigation than in the pastoral ecosystem (OR: 0.13; 95%CI 0.07, 0.23). Children living in areas with health care facilities had a low odd of malaria infection by 45% (OR: 0.55; 95% CI = 0.35, 0.86). Overall, the prevalence of anaemia in the district was 43.4% (n = 775); and 58.3% of those with severe anaemia were among children from the pastoral villages. Anaemia was significantly higher among children not using mosquito nets (p = 0.049); and among those with malaria infection (p <0.001). The majority (96%) of the children had Body Mass Index less than 18.5 kg/m(2) which indicate high proportion of underweight. CONCLUSION: There are significant variations in the risk of acquiring malaria infection between different ecosystems and livelihoods. These findings suggest that malaria control programmes must take into account ecosystems and livelihoods of the targeted population through an integrated management of malaria and nutrition approach.

Social determinants of malaria and health care seeking patterns among rice farming and pastoral communities in Kilosa District in central Tanzania.

This study was carried out to understand the role social determinants and health seeking behavior among rice farming and pastoral communities in Kilosa District in central Tanzania. The study involved four villages; two with rice farming communities while the other two with pastoral communities. In each village, heads of households or their spouses were interviewed to seek information on livelihoods activities, knowledge and practices on malaria and its preventions. A total of 471 individuals (males=38.9%; females=61.1%) were interviewed. Only 23.5% of the respondents had adequate knowledge on malaria. Fifty-six percent of the respondents could not associate any livelihood activity with malaria transmission. Majority (79%) of the respondents believed that most of fevers were due to malaria; this was higher among the pastoral (81.7%) than rice farming communities (76.1%) (p=0.038). Cases of fever were significantly higher in households with non-educated (31.2%) than educated respondents (21.5%). Women experienced significantly more episodes of fever than men (p<0.001). Of the total of 2606 individuals living in the households, 26.9% were reported to have had fever in the previous three months. Fever was reported more frequently among pastoral than rice farming communities (p<0.01). Of those who had fever, 36.6% were clinically diagnosed with malaria and 22.9% were confirmed to be infected with malaria. A combination of fever+convulsions or joint pains+headache was most frequently perceived to be malaria. Treatment seeking frequency differed by the size of the household and between rice farming and pastoral communities (p=0.05). In conclusion, education, sex, availability of health care facility and livelihood practices were the major social determinants that influence malaria acquisition and care seeking pattern in central Tanzania. Appropriate public health promotion should be designed to address the links of livelihoods and malaria transmission among rural farming communities in an ecohealth approach.

Accessibility, availability and utilisation of malaria interventions among women of reproductive age in Kilosa district in central Tanzania.

BACKGROUND: Universal access to and utilization of malaria prevention measures is defined as every person at malaria risk sleeping under a quality insecticide-treated mosquito net (ITN) and every pregnant woman at risk receiving at least two doses of sulfadoxine-pyrimethamine (SP). This study aimed to determine factors affecting accessibility, availability and utilisation of malaria interventions among women of reproductive age in Kilosa district in central Tanzania. METHODS: Women of reproductive age with children <5 years old or those who had been pregnant during the past 5 years were included in the study. A structured questionnaire was used to seek information on malaria knowledge, accessibility and utilization of malaria interventions during pregnancy. RESULTS: A total of 297 women (mean age=29±6.8 years) were involved. Seventy percent of the women had attained primary school education. About a quarter of women had two children of <5 years while over 58% had ≥3 children. Most (71.4%) women had medium general knowledge on malaria while only eight percent of them had good knowledge on malaria in pregnancy. A significant proportion of women were not aware of the reasons for taking SP during pregnancy (35%), timing for SP (18%), and the effect of malaria on pregnancy (45.8%). Timing for first dose of SP for intermittent preventive treatment in pregnancy (IPTp) was 1-3 months (28.4%) and 4-6 months (36.8%). Some 78.1% were provided with SP under supervision of the health provider. Knowledge on malaria in pregnancy had a significant association with levels of education (p=0.024). Ninety-eight percent had an ITN, mostly (87.1%) received free from the government. All women attended the ANC during their last pregnancy. The coverage of IPT1 was 53.5% and IPTp2 was 41.1%. The proportion of women making more ANC visits decreased with increasing parity. CONCLUSION: This study showed that the knowledge of the pregnant women on malaria in pregnancy and IPTp was average and is likely to have an impact on the low IPTp coverage. Campaigns that provide educational massages on the risk of malaria during pregnancy and the usefulness of IPTp need to be emphasised.

Impact of indoor residual spraying of lambda-cyhalothrin on malaria prevalence and anemia in an epidemic-prone district of Muleba, north-western Tanzania.

The Government of Tanzania introduced indoor residual spraying (IRS) in Muleba district in north-western Tanzania after frequent malaria epidemics. Malaria parasitological baseline and two cross-sectional follow-up surveys were conducted in villages under the IRS program and those not under IRS to assess the impact of IRS intervention. After two rounds of IRS intervention there was a significant reduction of malaria parasitological indices in both two villages. In IRS villages overall, parasitemia prevalence was reduced by 67.2%, splenomegaly was reduced by 75.8%, whereas anemia was reduced by 50.5%. There was also a decline of malaria parasite density from 896.4 at baseline to 128.8 at second follow-up survey. Similarly, there was also a reduction of malaria parasitological indices in non-IRS villages; however, parasitological indices in IRS villages remained far below the levels in non-IRS villages. The reduction of malaria parasitological indices in non-IRS villages might have been contributed by interventions other than IRS.

Species shifts in the Anopheles gambiae complex: do LLINs successfully control Anopheles arabiensis?

INTRODUCTION: High coverage of conventional and long-lasting insecticide treated nets (ITNs and LLINs) in parts of E Africa are associated with reductions in local malaria burdens. Shifts in malaria vector species ratio have coincided with the scale-up suggesting that some species are being controlled by ITNs/LLINs better than others. METHODS: Between 2005-2006 six experimental hut trials of ITNs and LLINs were conducted in parallel at two field stations in northeastern Tanzania; the first station was in Lower Moshi Rice Irrigation Zone, an area where An. arabiensis predominates, and the second was in coastal Muheza, where An. gambiae and An. funestus predominate. Five pyrethroids and one carbamate insecticide were evaluated on nets in terms of insecticide-induced mortality, blood-feeding inhibition and exiting rates. RESULTS: In the experimental hut trials mortality of An. arabiensis was consistently lower than that of An. gambiae and An. funestus. The mortality rates in trials with pyrethroid-treated nets ranged from 25-52% for An. arabiensis, 63-88% for An. gambiae s.s. and 53-78% for An. funestus. All pyrethroid-treated nets provided considerable protection for the occupants, despite being deliberately holed, with blood-feeding inhibition (percentage reduction in biting rates) being consistent between species. Veranda exiting rates did not differ between species. Percentage mortality of mosquitoes tested in cone bioassays on netting was similar for An. gambiae and An. arabiensis. CONCLUSIONS: LLINs and ITNs treated with pyrethroids were more effective at killing An. gambiae and An. funestus than An. arabiensis. This could be a major contributing factor to the species shifts observed in East Africa following scale up of LLINs. With continued expansion of LLIN coverage in Africa An. arabiensis is likely to remain responsible for residual malaria transmission, and species shifts might be reported over larger areas. Supplementary control measures to LLINs may be necessary to control this vector species.

Malaria risk factors in north-east Tanzania.

BACKGROUND: Understanding the factors which determine a household's or individual's risk of malaria infection is important for targeting control interventions at all intensities of transmission. Malaria ecology in Tanzania appears to have reduced over recent years. This study investigated potential risk factors and clustering in face of changing infection dynamics. METHODS: Household survey data were collected in villages of rural Muheza district. Children aged between six months and thirteen years were tested for presence of malaria parasites using microscopy. A multivariable logistic regression model was constructed to identify significant risk factors for children. Geographical information systems combined with global positioning data and spatial scan statistic analysis were used to identify clusters of malaria. RESULTS: Using an insecticide-treated mosquito net of any type proved to be highly protective against malaria (OR 0.75, 95% CI 0.59-0.96). Children aged five to thirteen years were at higher risk of having malaria than those aged under five years (OR 1.71, 95% CI 1.01-2.91). The odds of malaria were less for females when compared to males (OR 0.62, 95% CI 0.39-0.98). Two spatial clusters of significantly increased malaria risk were identified in two out of five villages. CONCLUSIONS: This study provides evidence that recent declines in malaria transmission and prevalence may shift the age groups at risk of malaria infection to older children. Risk factor analysis provides support for universal coverage and targeting of long-lasting insecticide-treated nets (LLINs) to all age groups. Clustering of cases indicates heterogeneity of risk. Improved targeting of LLINs or additional supplementary control interventions to high risk clusters may improve outcomes and efficiency as malaria transmission continues to fall under intensified control.

An experimental hut evaluation of Olyset nets against anopheline mosquitoes after seven years use in Tanzanian villages.

BACKGROUND: Long-lasting insecticidal nets (LLINs) are advocated by WHO for protection against malaria. Of the three brands of LLINs currently approved by WHO, Olyset(R) is the only one currently granted full recommendation. With this type of LLIN, the insecticide (permethrin) is incorporated into the polyethylene fibre during manufacture and diffuses from the core to the surface, thereby maintaining surface concentrations. It has not been determined for how long Olyset nets remain protective against mosquitoes in household use. METHODS: Examples of Olyset nets, which had been in use in Tanzanian villages for seven years, were tested in experimental huts against naturally entering Anopheles gambiae and Anopheles funestus mosquitoes. Performance was compared with new Olyset nets, conventionally treated ITNs (either newly treated with alphacypermethrin or taken from local villages after 1.5 years of use) and untreated nets. All nets were artificially holed except for the seven-year Olyset nets, which had developed holes during prolonged domestic use. RESULTS: Anopheles funestus and An. gambiae in NE Tanzania are susceptible to pyrethroids. The new Olyset nets caused high mortality against An. funestus (73.9%) and An. gambiae (62.7%) in experimental huts. The seven-year Olyset nets caused 58.9% mortality against An. funestus and 40.0% mortality against An. gambiae. The freshly treated alphacypermethrin nets also caused high mortality against An. funestus (70.6%) and An. gambiae (72.0%); this decreased to 58.4% and 69.6% respectively after 1.5 years of use. The new Olyset nets inhibited blood-feeding by 40-50%. The 7 year Olyset nets showed no feeding inhibition over that shown by the untreated nets. The alphacypermethrin treated nets failed to inhibit blood-feeding after 1.5 years of use. However iHhhdn laboratory tunnel tests samples of all types of treated net including the 7 year Olyset inhibited blood-feeding by more than 95%. CONCLUSION: After seven years of use Olyset nets were still strongly insecticidal. Mosquito mortality decreased by only 20-35% over this period. However, Olyset would not provide personal protection after seven years unless it was in good condition and all holes fully repaired.