Farmers' pesticide usage practices in the malaria endemic region of North-Western Tanzania: implications to the control of malaria vectors.

BACKGROUND: Pesticides remain the mainstay for the control of agricultural pests and disease vectors. However, their indiscriminate use in agriculture has led to development of resistance to both crop pests and disease vectors. This threatens to undermine the success gained through the implementation of chemical based vector control programs. We investigated the practices of farmers with regard to pesticide usage in the vegetable growing areas and their impact on susceptibility status of An. gambiae s.l. METHODS: A stratified multistage sampling technique using the administrative structure of the Tanzanian districts as sampling frame was used. Wards, villages and then participants with farms where pesticides are applied were purposively recruited at different stages of the process, 100 participants were enrolled in the study. The same villages were used for mosquito larvae sampling from the farms and the surveys were complimented by the entomological study. Larvae were reared in the insectary and the emerging 2-3 days old female adults of Anopheles gambiae s.l were subjected to susceptibility test. RESULTS: Forty eight pesticides of different formulations were used for control of crop and Livestock pests. Pyrethroids were the mostly used class of pesticides (50%) while organophosphates and carbamates were of secondary importance. Over 80% of all farmers applied pesticides in mixed form. Susceptibility test results confirmed high phenotypic resistance among An. gambiae populations against DDT and the pyrethroids (Permethrin-0.75%, Cyfluthrin-0.15%, Deltametrin-0.05% and Lambdacyhalothrin-0.05%) with mortality rates 54, 61, 76 and 71%, respectively. Molecular analysis showed An. arabiensis as a dominant species (86%) while An. gambiae s.s constituted only 6%. The kdr genes were not detected in all of the specimens that survived insecticide exposures. CONCLUSION: The study found out that there is a common use of pyrethroids in farms, Livestocks as well as in public health. The study also reports high phenotypic resistance among An. gambiae s.l against most of the pyrethroids tested. The preponderance of pyrethroids in agriculture is of public health concern because this is the class of insecticides widely used in vector control programs and this calls for combined integrated pest and vector management (IPVM).

Validation of droplet digital Polymerase Chain Reaction for the detection and absolute quantification of Taenia solium eggs in spiked soil samples.

To enable the detection of taeniid eggs in environmental samples, a sensitive technology is required. In this study, we validated the effectiveness of a digital droplet Polymerase Chain Reaction (ddPCR) assay for detection, identification and absolute quantification of taeniid DNA from artificially contaminated soils with varying numbers of taeniid eggs using a set of universal primers, JB3 & JB4.5. The results showed that the number of cox1 copies detected increased gradually for both species with the number of taeniid eggs added to the different soil types. The highest cox1 DNA copies recovery for Taenia solium and T. lynciscapreoli was from the sand soil with lowest recovery being observed in clay soils. Therefore, ddPCR is a promising technology for screening of taeniid eggs from soil samples collected in the environment irrespective of the soil type and the number of eggs. The potential of the ddPCR protocol to detect taeniid egg DNA in spiked soil samples has great practical application for taeniid egg screening in soils from endemic areas. However, when universal primers are used in screening environmental samples, the identity of ddPCR positive samples must be confirmed by sequencing. In addition, more validation studies using species-specific primers and field soil samples is recommended.

The status and health burden of neurocysticercosis in Mbulu district, northern Tanzania.

OBJECTIVE: The objective of this study was to assess the extent and health burden of neurocysticercosis in the general community of the Mbulu district, northern Tanzania. About 1051 randomly select participants were screened for human cysticercosis. The Cysticercus Western Blot IgG and Computed Tomography scan were used to detect infection by cysticerci. The DALYs was used to assess the community's health burden vis-a-vis neurocysticercosis. RESULTS: The sero-prevalence of HCC was 16.27%. About 76% of 25 selected human cysticercosis sero-positives had neurocysticercosis suggestive lesions on CT scan and 74% had history of epilepsy. Epilepsy caused 2.8 years of life lost and 2.2 healthy years of life lost due to disability per 1000 person-years in Mbulu. The average DALYs imposed due to neurocysticercosis and epilepsy were 3.0 and 3.9 per 1000 person-years, respectively. Neurocysticercosis is a serious public health concern in northern Tanzania.

Assessing S. mansoni prevalence in Biomphalaria snails in the Gombe ecosystem of western Tanzania: the importance of DNA sequence data for clarifying species identification.

BACKGROUND: Snails are essential for the transmission and maintenance of schistosomiasis in endemic areas, as they serve as intermediate hosts for schistosome parasites. A clear understanding of the snail species present, their local distribution and infection status is therefore a prerequisite for effective control of schistosomiasis. The purpose of this study was to establish the infection status and distribution of Schistosoma mansoni in snails in the Gombe area along the shores of Lake Tanganyika in western Tanzania, using both detection of cercarial shedding and molecular approaches. METHODS: Snails were collected from streams located close to human settlements in Gombe National Park, as well as from nearby villages (Kiziba, Mtanga, Mwamgongo and Bugamba) and the largest town in the region (Kigoma). Snails were individually exposed to light to induce shedding of schistosome larvae, which were examined using a compound light microscope. Additionally, the internal transcribed spacer (ITS) region of the ribosomal RNA gene cluster was simultaneously amplified in both snails and their trematodes using a single polymerase chain reaction (PCR) and sequenced to confirm species identification. RESULTS: Snails morphologically identified as Biomphalaria pfeifferi were present in all streams except at Mtanga but their distribution was patchy in both time and space. Sequencing of PCR products indicated that not all snails were B. pfeifferi. None of the snails from Gombe or Bugamba shed schistosome larvae, while larvae were shed at all other sites. Overall, an infection prevalence of only 12% was observed in snails based on cercarial shedding. While 47% of the snails were PCR-positive for the 500 bp ITS fragment, which was predicted to indicate infection with S. mansoni, sequence data demonstrated that these bands are not species-specific and can be amplified from other trematode infections. In addition, a 1000 bp band was amplified in 14% of samples, which was identified as a trematode in the family Derogenidae. CONCLUSIONS: The results support the previous assumption that B. pfeifferi snails may be involved in transmitting schistosomiasis in the area but suggest that the community structure of both snails and trematodes may be more complicated than previously thought. This emphasises the importance of confirming species identifications using sequencing, rather than relying only on PCR-based diagnostics or cercarial shedding.

Estimating the prevalence and intensity of Schistosoma mansoni infection among rural communities in Western Tanzania: The influence of sampling strategy and statistical approach.

BACKGROUND: Schistosoma mansoni is a parasite of major public health importance in developing countries, where it causes a neglected tropical disease known as intestinal schistosomiasis. However, the distribution of the parasite within many endemic regions is currently unknown, which hinders effective control. The purpose of this study was to characterize the prevalence and intensity of infection of S. mansoni in a remote area of western Tanzania. METHODOLOGY/PRINCIPAL FINDINGS: Stool samples were collected from 192 children and 147 adults residing in Gombe National Park and four nearby villages. Children were actively sampled in local schools, and adults were sampled passively by voluntary presentation at the local health clinics. The two datasets were therefore analysed separately. Faecal worm egg count (FWEC) data were analysed using negative binomial and zero-inflated negative binomial (ZINB) models with explanatory variables of site, sex, and age. The ZINB models indicated that a substantial proportion of the observed zero FWEC reflected a failure to detect eggs in truly infected individuals, meaning that the estimated true prevalence was much higher than the apparent prevalence as calculated based on the simple proportion of non-zero FWEC. For the passively sampled data from adults, the data were consistent with close to 100% true prevalence of infection. Both the prevalence and intensity of infection differed significantly between sites, but there were no significant associations with sex or age. CONCLUSIONS/SIGNIFICANCE: Overall, our data suggest a more widespread distribution of S. mansoni in this part of Tanzania than was previously thought. The apparent prevalence estimates substantially under-estimated the true prevalence as determined by the ZINB models, and the two types of sampling strategies also resulted in differing conclusions regarding prevalence of infection. We therefore recommend that future surveillance programmes designed to assess risk factors should use active sampling whenever possible, in order to avoid the self-selection bias associated with passive sampling.

The influence of age on insecticide susceptibility of Anopheles arabiensis during dry and rainy seasons in rice irrigation schemes of Northern Tanzania.

BACKGROUND: Insecticide resistance is the major emerging challenge facing the malaria vector control programmes in Tanzania. Proper monitoring and detection is of paramount importance guiding the vector control programmes. This paper presents the effect of mosquito aging on insecticide resistance status in Anopheles arabiensis populations in dry and rainy seasons in northern Tanzania. METHODS: Anopheles gambiae s.l. larvae were sampled from rice fields in both dry and rainy seasons and reared in the insectary to adults. The emerged females in batches of 2, 3, 5, and 10 days old were exposed to six insecticides (deltamethrin, permethrin, lambda-cyhalothrin, DDT, bendiocarb and pirimiphos-methyl) to see the effects of age on insecticide resistance. Mosquitoes were exposed to insecticides using WHO standard susceptibility test kits. Knockdown was recorded during the 1-h exposure, while mortality and resistance ratio were recorded 24 h later. Mosquito specimens were identified to species level using the polymerase chain reaction (PCR) method. RESULTS: Among the 326 specimens processed by PCR, 323 (99.1%) were identified as Anopheles arabiensis. There was reduced mortality (ranging from 61 to 97.7%) when adults reared from larvae were exposed to all pyrethroids and bendiocarb in both dry and rainy seasons, while they were fully susceptible to DDT and pirimiphos-methyl. There was a significant increase in mortality rate with increase in mosquito's age in both dry and rainy seasons following exposure to pyrethroids (DF = 1, P < 0.05). Mosquitoes showed significantly higher mortality rates in the rainy season than in the dry season after being exposed to pyrethroids (DF = 1, P < 0.05). Higher mortality rates (94.0-99.8%) were observed in all ages and seasons when mosquitoes were exposed to bendiocarb compared with pyrethroids. Pirimiphos-methyl was only tested in the rainy season so no comparison with dry season mosquitoes could be made. CONCLUSIONS: Results showed that An. arabiensis were resistant to pyrethroids in both seasons and that the young age groups exhibited higher levels of resistance compared with the older age groups. Mosquitoes were full susceptible to DDT and pirimiphos-methyl irrespective of the season and age.

Genetic diversity of Glossina fuscipes fuscipes along the shores of Lake Victoria in Tanzania and Kenya: implications for management.

BACKGROUND: Tsetse flies (Diptera: Glossinidae) are sole vectors for trypanosomiasis, which affect human health and livestock productivity in Africa. Little is known about the genetic diversity of Glossina fuscipes fuscipes, which is an important species in Tanzania and Kenya. The main objective of the study was to provide baseline data to determine the genetic variability and divergence of G. f. fuscipes in the Lake Victoria basin of Tanzania and Kenya in order to guide future vector control efforts in the region. FINDINGS: Two hundred and seventy five G. f. fuscipes from 8 sites along the shores of Lake Victoria were screened for genetic polymorphisms at 19 microsatellite loci. Samples were collected from two sites in Kenya and six sites in Tanzania. Four of the Tanzanian sites were located in the Rorya district, on the eastern shores of Lake Victoria, while the other two sites were from Ukerewe and Bukoba districts from the southern and western Lake Victoria shores, respectively. Four genetically distinct allopatric clusters were revealed by microsatellite analysis, which sorted the sampling sites according to geography, with sites separated by as little as ~65 km belonging to distinct genetic clusters, while samples located within ~35 km from each other group in the same cluster. CONCLUSION: Our results suggest that there is ongoing genetic admixture within sampling sites located ~35 km from each other, while sites located ~65 km apart are genetically isolated from each other. Similar patterns emerged from a parallel study on G. f. fuscipes analyzed from the Lake Victoria Uganda shores. From a control perspective these results suggest that for sites within the same genetic cluster, control efforts should be carried out in a coordinated fashion in order to avoid re-invasions. Future work should focus on better quantifying the extent and spatial patterns of the observed genetic discontinuities of the G. f. fuscipes populations along the Tanzanian shores. This will aid in their control by providing guidelines on the geographical extent of the area to be treated at the same time.

Habitat productivity and pyrethroid susceptibility status of Aedes aegypti mosquitoes in Dar es Salaam, Tanzania.

BACKGROUND: Aedes aegypti (Diptera: Culicidae) is the main vector of the dengue virus globally. Dengue vector control is mainly based on reducing the vector population through interventions, which target potential breeding sites. However, in Tanzania, little is known about this vector's habitat productivity and insecticide susceptibility status to support evidence-based implementation of control measures. The present study aimed at assessing the productivity and susceptibility status of A. aegypti mosquitoes to pyrethroid-based insecticides in Dar es Salaam, Tanzania. METHODS: An entomological assessment was conducted between January and July 2015 in six randomly selected wards in Dar es Salaam, Tanzania. Habitat productivity was determined by the number of female adult A. aegypti mosquitoes emerged per square metre. The susceptibility status of adult A. aegypti females after exposure to 0.05% deltamethrin, 0.75% permethrin and 0.05% lambda-cyhalothrin was evaluated using the standard WHO protocols. Mortality rates were recorded after 24 h exposure and the knockdown effect was recorded at the time points of 10, 15, 20, 30, 40, 50 and 60 min to calculate the median knockdown times (KDT50 and KDT95). RESULTS: The results suggest that disposed tyres had the highest productivity, while water storage tanks had the lowest productivity among the breeding habitats Of A. aegypti mosquitoes. All sites demonstrated reduced susceptibility to deltamethrin (0.05%) within 24 h post exposure, with mortalities ranging from 86.3 ± 1.9 (mean ± SD) to 96.8 ± 0.9 (mean ± SD). The lowest and highest susceptibilities were recorded in Mikocheni and Sinza wards, respectively. Similarly, all sites demonstrated reduced susceptibility permethrin (0.75%) ranging from 83.1 ± 2.1% (mean ± SD) to 96.2 ± 0.9% (mean ± SD), in Kipawa and Sinza, respectively. Relatively low mortality rates were observed in relation to lambda-cyhalothrin (0.05%) at all sites, ranging from 83.1 ± 0.7 (mean ± SD) to 86.3 ± 1.4 (mean ± SD). The median KDT50 for deltamethrin, permethrin and lambda-cyhalothrin were 24.9-30.3 min, 24.3-34.4 min and 26.7-32.8 min, respectively. The KDT95 were 55.2-90.9 min for deltamethrin, 54.3-94.6 min for permethrin and 64.5-69.2 min for lambda-cyhalothrin. CONCLUSIONS: The productive habitats for A. aegypti mosquitoes found in Dar es Salaam were water storage containers, discarded tins and tyres. There was a reduced susceptibility of A. aegypti to and emergence of resistance against pyrethroid-based insecticides. The documented differences in the resistance profiles of A. aegypti mosquitoes warrants regular monitoring the pattern concerning resistance against pyrethroid-based insecticides and define dengue vector control strategies.

Pyrethroids and DDT tolerance of Anopheles gambiae s.l. from Sengerema District, an area of intensive pesticide usage in north-western Tanzania.

OBJECTIVE: To assess the susceptibility status of malaria vectors to pyrethroids and dichlorodiphenyltrichloroethane (DDT), characterise the mechanisms underlying resistance and evaluate the role of agro-chemical use in resistance selection among malaria vectors in Sengerema agro-ecosystem zone, Tanzania. METHODS: Mosquito larvae were collected from farms and reared to obtain adults. The susceptibility status of An. gambiae s.l. was assessed using WHO bioassay tests to permethrin, deltamethrin, lambdacyhalothrin, etofenprox, cyfluthrin and DDT. Resistant specimens were screened for knock-down resistance gene (kdr), followed by sequencing both Western and Eastern African variants. A gas chromatography-mass spectrophotometer (GC-MS) was used to determine pesticide residues in soil and sediments from mosquitoes' breeding habitats. RESULTS: Anopheles gambiae s.l. was resistant to all the insecticides tested. The population of Anopheles gambiae s.l was composed of Anopheles arabiensis by 91%. The East African kdr (L1014S) allele was found in 13 of 305 specimens that survived insecticide exposure, with an allele frequency from 0.9% to 50%. DDTs residues were found in soils at a concentration up to 9.90 ng/g (dry weight). CONCLUSION: The observed high resistance levels of An. gambiae s.l., the detection of kdr mutations and pesticide residues in mosquito breeding habitats demonstrate vector resistance mediated by pesticide usage. An integrated intervention through collaboration of agricultural, livestock and vector control units is vital.

Extent of morbidity associated with schistosomiasis infection in Malawi: a review paper.

Data on the extent of the burden due to schistosomiasis is sparse in most Sub-Saharan African countries. However, this data is crucial for triggering medical attention. A review of extent of morbidity and determinants associated with schistosomiasis in Malawi was therefore conducted to quantify the infection in order to concretise the need for medical intervention. A systematic and traditional search strategy was used to find literature for the review, whilst exclusion and inclusion criteria were used to identify appropriate articles. Logistic regression curves of epidemiological model Y = (a + bx (c) )/(1 + bx (c) ) and the recommendation that schistosomiasis prevalence can be used to estimate morbidity were employed to quantify morbidity at various infection stages. Morbidity was quantified as a direct proportion of the population and the respective national schistosomiasis prevalence. Findings showed that both S. mansoni and S. haematobium are present in Malawi with the latter highly prevalent (50%). Furthermore, out of the estimated population of 16,829 million, approximately 8.4 million have schistosomiasis, with about 4.4 million of these aged 18 years and below. The most frequent manifestation is Katayama syndrome, while ascites is the lowest, impacting about 3.0 million and 960 individuals, respectively. Localised studies on association of schistosomiasis infection to risk factors such as occupation, age and gender found odds ratio (OR) ranging from 1.29 to 5.37. Morbidity due to schistosomiasis is high in Malawi. It is therefore recommended that a more detailed study on the determinants of high schistosomiasis and re-evaluation of the current control measures be conducted if the current morbidity statistics are to be remarkably reduced.

The impact of livestock on the abundance, resting behaviour and sporozoite rate of malaria vectors in southern Tanzania.

BACKGROUND: Increases in the coverage of long-lasting insecticidal nets (LLINs) have significantly reduced the abundance of Anopheles gambiae sensu stricto in several African settings, leaving its more zoophagic sibling species Anopheles arabiensis as the primary vector. This study investigated the impact of livestock ownership at the household level on the ecology and malaria infection rate of vectors in an area of Tanzania where An. arabiensis accounts for most malaria transmission. METHODS: Mosquito vectors were collected resting inside houses, animal sheds and in outdoor resting boxes at households with and without livestock over three years in ten villages of the Kilombero Valley, Tanzania. Additionally, the abundance and sporozoite rate of vectors attempting to bite indoors at these households was assessed as an index of malaria exposure. RESULTS: The mean abundance of An. gambiae s.l. biting indoors was similar at houses with and without livestock. In all years but one, the relative proportion of An. arabiensis within the An. gambiae s.l. species complex was higher at households with livestock. Livestock presence had a significant impact on malaria vector feeding and resting behaviour. Anopheles arabiensis were generally found resting in cattle sheds where livestock were present, and inside houses when absent. Correspondingly, the human blood index of An. arabiensis and An. funestus s.l. was significant reduced at households with livestock, whereas that of An. gambiae s.s. was unaffected. Whilst there was some evidence that sporozoite rates within the indoor-biting An. gambiae s.l population was significantly reduced at households with livestock, the significance of this effect varied depending on how background spatial variation was accounted for. CONCLUSIONS: These results confirm that the presence of cattle at the household level can significantly alter the local species composition, feeding and resting behaviour of malaria vectors. However, the net impact of this livestock-associated variation in mosquito ecology on malaria exposure risk was unclear. Further investigation is required to distinguish whether the apparently lower sporozoite rates observed in An. gambiae s.l. at households with livestock is really a direct effect of cattle presence, or an indirect consequence of reduced risk within areas where livestock keepers choose to live.

Direct and indirect effect of predators on Anopheles gambiae sensu stricto.

The increased insecticides resistance by vectors and the ecological harm imposed by insecticides to beneficial organisms drawback mosquitoes chemical control efforts. Biological control would reduce insecticides tolerance and yet biodiversity friend. The predatory and non-predatory effects of Gambusia affinis and Carassius auratus on gravid Anopheles gambiae sensu strict and larvae survivorship were assessed. In determining predation rate, a single starved predator was exposed to third instar larvae of An. gambiae s.s. in different densities 20, 60 and 100. Six replicates in each of the densities for both predators, G. affinis and C. auratus, were set up. The larvae densities were monitored in every 12 and 24 h. In assessing indirect effects: An. gambiae s.s. first instar larvae of three densities 20, 60 and 100 were reared in water from a predator habitat and water from non-predator habitat. Larvae were monitored until they emerged to adults where larval survivorship and sex ratio (Female to total emerged mosquitoes) of the emerged adult from both water habitats were determined. Oviposition preference: twenty gravid females of An. gambiae s.s. were provided with three oviposition choices, one containing water from predator habitat without a predator, the second with water from a predator with a predator and the third with water from non-predatory habitat. The number of eggs laid on each container was counted daily. There were 20 replicates for each predator, G. affinis and C. auratus. Survivorship of An. gambiae s.s. larvae reared in water from non-predator habitat was higher than those reared in water from the predator habitats. Many males emerged in water from non-predatory water habitats while more females emerged from predator's habitats water. More eggs laid in tap water than in water from predator habitat and water from predator habitat with live predator. In 24 h, a starved C. auratus and G. affinis were able to consume 100% of the 3rd instar larvae. The findings from this study suggest that G. affinis and C. auratus may be useful in regulating mosquito populations in favour of beneficial insects. However, a small scale trial shall be needed in complex food chain system to ascertain the observed predation and kairomones effects.

Larvicidal efficacy of Cryptomeria japonica leaf essential oils against Anopheles gambiae.

BACKGROUND: Alternative insecticidal compounds with mortality effect against mosquito life cycle stages are currently needed. The compounds should be biodegradable and nontoxic to non-targeted insects. Plant based larvicides provide effective control of vector populations. This study explored Cryptomeria japonica leaf essential oil larvicidal potency against Anopheles gambiae sensu stricto. METHODS: Essential oils (12.5 to 200 µg/mL) extracted from C. japonica leaves were evaluated against An. gambiae s.s. larvae in both the laboratory and semi field in 6 replicates for each dose. Larval mortality readings were taken at 12, 24, 48, and 72 h post treatment. RESULTS: C. japonica leaf essential oil yield was 17.06 ± 0.56 mL/kg and 1.60 ± 0.33% (w/w). GC-FID and GC-MS analyses revealed 22 constituents. Essential oil was more effective against An. gambiae s.s. larvae in the laboratory than in semi field trials. Mortality increased with increasing dosages (12.5 to 200 µg/mL) in the laboratory (31.75 to 100%) and semi field trials (17.75 to 99.5%), respectively. The LC50 value ranged from 5.55 to 63.92 µg/mL in the laboratory, and 8.22 to 134.84 µg/mL in semi field conditions, LC90 value ranged from 41.34 to 205.93 µg/mL in the laboratory and 50.92 to 213.11 µg/mL in semi field conditions. CONCLUSION: This study has demonstrated the potential of C. japonica leaf essential oil to cause mortality effects to An. gambiae s. s. larval populations, however, further studies need to be conducted under field conditions and also with individual active compounds of C. japonica essential oil.

The risk factors for human cysticercosis in Mbulu District, Tanzania.

The objective of this study was to explore the reasons for the persistence of human cysticercosis (HCC) transmission in Mbulu District, northern Tanzania. The study was carried out in 25 villages, whereby five major risks were identified. The risks were indiscriminate defaecation and improper use of toilets; a free-range system of keeping pigs; indiscriminate or unregulated slaughtering and inadequate meat hygiene and inspection; consumption of undercooked and porcine cysticerci infected pork; and social structure and roles. All of the identified risks were backed up by the immanent lifestyles of the community involved. These findings are important for the development of intervention strategies in the study area.

Larvicidal effect of disinfectant soap on Anopheles gambiae s.s (Diptera: Culicidae) in laboratory and semifield environs.

BACKGROUND: Mosquito larval control using chemicals and biological agents is of paramount importance in vector population and disease incidence reduction. A commercial synthetic disinfectant soap was evaluated against larvae of Anopheles gambiae s.s. in both laboratory and semi field conditions. METHOD: Five concentrations of commercial synthetic disinfectant soap (0.0001, 0.001, 0.01, 0.1 and 1%) were prepared and evaluated against third instar larvae in laboratory and semi field environments. Mortality was scored at 12, 24, 48, and 72 hrs. Each dosage had 6 replicates, having twenty 3rd instar larvae of An.gambiae s.s. RESULTS: In the laboratory phase, all dosages had significantly higher larval mortalities than in controls, while in semi field conditions, the dosages of 0.0001, 0.001 and 0.01% had lower mortalities than laboratory trials. In the comparison between semi field and laboratory trials, only 0.1 and 1% dosage had significant difference with more mortality in semifield conditions. Proportions of larvae that died during mortality monitoring intervals in laboratory and semi field had significant differences only at 12 hrs and 72 hrs. CONCLUSION: The findings of this study have demonstrated that the mortality of larvae caused by commercial synthetic disinfectant soap is worth further studies in open water bodies. More studies are necessary to find out the effect of sunlight on the chemistry of the synthetic disinfectant and other variables in small scale full field trials.

Activity of Cinnamomum osmophloeum leaf essential oil against Anopheles gambiae s.s.

BACKGROUND: The increasing status of insecticide resistant mosquitoes in sub-Saharan Africa is a threatening alert to the existing control efforts. All sibling species of An. gambiae complex have evolved insecticide resistance in wild populations for different approved classes of the insecticides currently in use in the field. An alternative compound for vector control is absolutely urgently needed. In this study, the larvicidal activity and chemical composition of the Cinnamomum osmophloeum leaf essential oils were investigated. METHODS: C. osmophloeum leaf essential oils were extracted by hydrodistillation in a Clevenger-type apparatus for 6 hours, and their chemical compositions identified using GC-MS. These oils were evaluated against An. gambiae s.s. in both laboratory and semi-field situations. The WHO test procedures for monitoring larvicidal efficacy in malaria vectors were used. RESULTS: The composition of C. osmophloeum leaf essential oil has been found to have 11 active compounds. The most abundant compound was trans-cinnamaldehyde (70.20%) and the least abundant was caryophyllene oxide (0.08%). The larvicidal activity was found to be dosage and time dependant both in laboratory and semi-field environments with mortality ranging from 0% to 100%. The LC50 value was found to vary from 22.18 to 58.15 µg/ml in the laboratory while in semi-field environments it was 11.91 to 63.63 µg/ml. The LC90 value was found to range between 57.71 to 91.54 µg/ml in the laboratory while in semi-field environments was 52.07 to 173.77 µg/ml. Mortality ranged from 13% to 100% in the laboratory while in semi-field environments it ranged between 43% to 100% within mortality recording time intervals of 12, 24, 48, and 72 hours. CONCLUSIONS: The larvicidal activity shown by C. osmophloeum leaf essential oil is a promising alternative to existing larvicides or to be incorporated in integrated larval source management compounds for An. gambiae s.s control. The efficacy observed in this study is attributed to both major and minor compounds of the essential oils.

A first insight into the barcodes for African diplostomids (Digenea: Diplostomidae): brain parasites in Clarias gariepinus (Siluriformes: Clariidae).

Diplostomid trematodes comprise a large and diverse group of widespread digeneans whose larval stages are important parasitic pathogens that may exert serious impacts in wild and cultured freshwater fish. However, our understanding of their diversity remains incomplete especially in the tropics. Our study is the first application of a DNA-based approach to diplostomid diversity in the African continent by generating a database linking sequences for the mitochondrial cytochrome c oxidase subunit 1 (cox1) barcode region and ITS1-5.8S-ITS2 rRNA gene cluster for brain-infecting diplostomid metacercariae from the catfish Clarias gariepinus. Analyses of newly-generated partial cox1 sequences for 34 larval isolates of Tylodelphys spp. from Tanzania and Diplostomum spp. from Tanzania and Nigeria revealed three strongly supported reciprocally monophyletic lineages of Tylodelphys spp. and one of an unknown species of Diplostomum. The average intraspecific divergence for the cox1 sequences for each recognised novel lineage was distinctly lower compared with interspecific divergence (0.46-0.75% vs 11.7-14.8%). The phylogenetic hypotheses estimated from Bayesian inference and maximum likelihood analyses of ITS1-5.8S-ITS2 data exhibited congruent strong support for the cox1-derived lineages. Our study thus provides molecular-based evidence for the existence of three distinct brain-infecting species co-occurring in natural populations of C. gariepinus. Based on phylogenetic analyses, we re-allocated Diplostomum mashonenseBeverley-Burton (1963) to the genus Tylodelphys as a new combination. We also generated cox1 and ITS1-5.8S-ITS2 sequences for an unknown species of Diplostomum from another African fish host, Synodontis nigrita.

Plasmodium falciparum msp1, msp2 and glurp allele frequency and diversity in sub-Saharan Africa.

BACKGROUND: The efficacy of anti-malarial drugs is assessed over a period of 28-63 days (depending on the drugs' residence time) following initiation of treatment in order to capture late failures. However, prolonged follow-up increases the likelihood of new infections depending on transmission intensity. Therefore, molecular genotyping of highly polymorphic regions of Plasmodium falciparum msp1, msp2 and glurp loci is usually carried out to distinguish recrudescence (true failures) from new infections. This tool has now been adopted as an integral part of anti-malarial efficacy studies and clinical trials. However, there are concerns over its utility and reliability because conclusions drawn from molecular typing depend on the genetic profile of the respective parasite populations, but this profile is not systematically documented in most endemic areas. This study presents the genetic diversity of P. falciparum msp1, msp2 and glurp markers in selected sub-Saharan Africa countries with varying levels of endemicity namely Malawi, Tanzania, Uganda, Burkina Faso and São Tomé. METHODS: A total 780 baseline (Day 0) blood samples from children less than seven years, recruited in a randomized controlled clinical trials done between 1996 and 2000 were genotyped. DNA was extracted; allelic frequency and diversity were investigated by PCR followed by capillary electrophoresis for msp2 and fragment sizing by a digitalized gel imager for msp1 and glurp. RESULTS AND CONCLUSION: Plasmodium falciparum msp1, msp2 and glurp markers were highly polymorphic with low allele frequencies. A total of 17 msp1 genotypes [eight MAD20-, one RO33- and eight K1-types]; 116 msp2 genotypes [83 3D7 and 33 FC27- types] and 14 glurp genotypes were recorded. All five sites recorded very high expected heterozygosity (HE) values (0.68 - 0.99). HE was highest in msp2 locus (HE=0.99), and lowest for msp1 (HE=0.68) (P<0.0001). The genetic diversity and allelic frequency recorded were independent of transmission intensity (P=0.84, P=0.25 respectively. A few genotypes had particularly high frequencies; however the most abundant showed only a 4% probability that a new infection would share the same genotype as the baseline infection. This is unlikely to confound the distinction of recrudescence from new infection, particularly if more than one marker is used for genotyping. Hence, this study supports the use of msp1, msp2 and glurp in malaria clinical trials in sub-Saharan Africa to discriminate new from recrudescent infections.

Spatial variability in the density, distribution and vectorial capacity of anopheline species in Rufiji district, south-eastern Tanzania.

Malaria transmission varies from one area to another and there are also local difference in time and space. The objective of the study was to determine the local variability of entomological parameters namely, mosquito abundance, human biting rate (HBR), sporozoite rate for Plasmodiumfalciparum and entomological inoculation rate (EIR). The study was carried out in Rufiji District south eastern Tanzania from October 2001 and September 2004. Adult mosquitoes were collected indoors by CDC light traps. PCR was employed to identify the species within the Anopheles gambiae complex. ELISA was used to determine the sporozoite rate. Over a three year sampling period a total of 64,875 female mosquitoes were caught using light-traps, and of these 28% were Anopheles gambiae complex, 25% An. funestus Giles, 1% An. pharoensis Theobald, 46% Culex species and the rest were Mansonia uniformis Theobald. Mosquito abundance and species composition varied seasonally, spatially and between years. Using PCR, three members of the Anopheles gambiae complex namely An. gambiae s.s. Giles (69%), An. arabiensis Paton (23%) and An. merus D6nitz (7%) were confirmed to occur in the study area. Plasmodiumfalciparum circumsporozoite antigen (CSA) rates were 3.5% for An. gambiae complex and 2.3% for An. funestus. The mean EIR ranged from 28-275 infective bites/person/year. Transmission indices varied over short distances, seasonally and between years. In conclusion, malaria transmission indices in the study area are one of the highest in Tanzania; and there is high variability of entomological parameters over a small geographical area.

Determinants of the parasite community of clariid fishes from Lake Victoria, Tanzania.

The factors that determine parasite assemblages among the clariid fishes of Lake Victoria, Tanzania were studied between August 2003 and February 2005. Six hundred and fifty-six fish belonging to seven species were necropsied and examined for parasites, from which 31 species of metazoan parasites were recorded. The community was dominated by the nematodes both in species and numbers. Most species were generalists with only two trematodes, Diplostomum mashonense and Tylodelphys species, being specialists of Clarias gariepinus. Ten species were considered core and predictable. Parasite species richness, number of individuals per host and Shannon-Wiener diversity indices were generally high. At the compound community level, a mean number of 7.8 parasites were shared among different species of fish and the maximum number of parasites species per fish at the infracommunity level was seven. Levels of similarity in parasite species richness at the component community level ranged from 29.6 to 61.5%. The study concludes that parasite communities in clariid fishes of Lake Victoria are structured by ecological factors. At the infracommunity level, host size, diet and vagility promoted a richer parasite community. At the compound level, two factors were crucial, namely the intermixing of the waters in the lake and the predominant and mobile C. gariepinus.