Prevalence of malaria parasitaemia in school children from two districts of Ghana earmarked for indoor residual spraying: a cross-sectional study.

BACKGROUND: Indoor residual spraying (IRS) is considered a valuable transmission control measure against malaria but exact efficacy data are not available for many epidemiological settings. This study was conducted to determine indicators for malaria epidemiology and transmission among school children as baseline assessment before IRS implementation in Ghana. METHODS: A cross-sectional study was conducted in Adansi South District of the Ashanti Region and Wa West District of the Upper West Region of Ghana. Malarial parasitaemia and anaemia were determined in pupils between the ages of 2 and 14 years from Early Childhood Development Centres and primary schools. Plasmodium falciparum parasitaemia was detected by light microscopy. RESULTS: Out of 1,649 pupils who were enrolled at participating schools, 684 were positive for plasmodia resulting in a baseline parasitaemia prevalence of 41.5%. Parasite rate was similar in the two districts (42.0% in Adansi South and 40.7% in Wa West), but differed across the nine sentinel schools ranging from 21 to 63% (p < 0.001). The mean haemoglobin concentration was 11.3 g/dl [standard deviation (SD) ±2.1]. Pupils who had moderate to mild anaemia (7.0-10.9 g/dl) constituted 41.7% of the study sample. CONCLUSION: The burden of parasitaemia, malaria and anaemia is a major public health problem among school children in rural Ghana with extensive heterogeneity between schools and warrants further investment in intervention measures.

Malaria transmission in two rural communities in the forest zone of Ghana.

Malaria transmission was assessed in two rural communities, Kona and Afamanaso in Sekyere South district, Ashanti Region, in the forest zone of Ghana to provide baseline data for ongoing clinical studies and the evaluation of the effect of interventions. Altogether, 3,479 Anopheles gambiae and 1,157 Anopheles funestus were caught by human landing catches. Sporozoite rates determined by either microscopy of salivary glands or enzyme-linked immunosorbent assay (ELISA) for Plasmodium falciparum in the two villages were 6.6% vs. 8.9% for the main vector A. gambiae and 3.2% vs. 6.3% for A. funestus. ELISA tests of dissected specimens compared to microscopy of salivary glands were 1.3 and 2.0 times more positive for A. gambiae and A. funestus, respectively. Plasmodium infections of 122 microscopically positive salivary glands of A. gambiae were identified by real-time PCR as 95 (77.9%) P. falciparum, 7 (5.7%) Plasmodium malariae, 7 (5.7%) Plasmodium ovale and 1 (0.8%) mixed infection of P. falciparum and P. malariae. Transmission in the area was found to be intense and perennial with some seasonal variations during the study period from Dec. 2003 to Aug. 2005. Although the two villages were only 10 km apart from each other, Annual Biting Rates (ABRs) and Annual Entomological Inoculation Rates (AEIRs) were much higher at Afamanaso (11,643 vs. 866) than at Kona (5,329 vs. 490). Most of the transmission (91.4%) occurred during bedtime hours from 21 to 6 h. It is important to note that there was still a substantial transmission before 21 h with AEIRs of 57.3 at Afamanso and 38.7 at Kona. The distribution of impregnated bednets alone, therefore, may not be sufficiently effective.

Microbiological diagnosis of polymicrobial periprosthetic joint infection revealed superiority of investigated tissue samples compared to sonicate fluid generated from the implant surface.

OBJECTIVES: In the microbiological diagnosis of periprosthetic joint infection (PJI), there is much discussion about the methodology of obtaining proper specimens, the processing technique, and suitable culture media. This retrospective study was conducted to analyse the accuracy of our culture techniques. METHODS: Tissue samples and components from 258 patients after revision arthroplasty of the hip, knee, and shoulder were investigated, and the results of tissue cultures (TC) were compared to those of sonicate fluid cultures (SFC). Furthermore, an evaluation was performed of the influence of different culture media on the detection rate. RESULTS: PJI was confirmed in 186 patients. The overall sensitivity of TC was no different to that of SFC (91.3% vs 90.8%, P = 1). In 153 cases (82.3%), TC and SFC showed concordant positive results. Results were discordant in 33 cases (17.7%). When differentiated according to the type of infection, TC showed significantly better results than SFC in detecting polymicrobial infections (97.0% vs 67.0%, P = 0.004). There were also significant differences between the culture media regarding the yield of microorganisms. CONCLUSIONS: TC was more effective in detecting co-infections. The best results were obtained using both TC and SFC. The choice of culture media has a significant influence on the quality of results.

Blood-feeding behavior of Anopheles gambiae and Anopheles melas in Ghana, western Africa.

Anopheles gambiae is the predominant malaria vector species in Ghana, western Africa, with a strong local presence of Anopheles melas Theobald along the southern coast. We studied the biting behavior of these two species of the Anopheles gambiae complex inland and at the coast in Ghana, with special attention to the local peoples' preference for outdoor sleeping. We collected mosquitoes at two sites in 2007, representing the moist semideciduous forest zone and the strand and mangrove zone, and the sampling was repeated in the dry and rainy seasons. Sampled mosquitoes were examined for species, parity and size (wing length), and we identified the hosts of their bloodmeals. We interviewed 288 of the village people to determine where and when they slept outdoors. Our study confirmed that An. gambiae is the only species of the An. gambiae complex in the Ashanti region and revealed that An. melas is highly dominant on the western coast of Ghana. Both species showed high human blood rates in indoor resting mosquito samples. More people sleep outside on the coast than inland. An. melas demonstrated high exophily. An. gambiae bit people more frequently indoors and did so more often during the dry season than in the rainy season. We suggest that the degree of exophily in An. melas may be affected by humidity and the availability of human as well as by the mosquitoes' innate habits.

Distant relatives of severe acute respiratory syndrome coronavirus and close relatives of human coronavirus 229E in bats, Ghana.

We tested 12 bat species in Ghana for coronavirus (CoV) RNA. The virus prevalence in insectivorous bats (n = 123) was 9.76%. CoV was not detected in 212 fecal samples from Eidolon helvum fruit bats. Leaf-nosed bats pertaining to Hipposideros ruber by morphology had group 1 and group 2 CoVs. Virus concentrations were < or =45,000 copies/100 mg of bat feces. The diversified group 1 CoV shared a common ancestor with the human common cold virus hCoV-229E but not with hCoV-NL63, disputing hypotheses of common human descent. The most recent common ancestor of hCoV-229E and GhanaBt-CoVGrp1 existed in approximately 1686-1800 ad. The GhanaBt-CoVGrp2 shared an old ancestor (approximately 2,400 years) with the severe acute respiratory syndrome-like group of CoV.

Evaluation of onchocerciasis control in the Upper Denkyira East municipal in the forest area of Ghana: Responses of participants and distributors to the CDTI programme.

The African Programme for Onchocerciasis Control (APOC), which focused on annual mass treatment with ivermectin, was launched in 1995 and was replaced by the Expanded Special Project for Neglected Tropical Diseases (ESPEN) by the end of 2015. In Ghana, the Community Directed Treatment with Ivermectin (CDTI) was introduced in 1999. After a decade, biannual reinforcement was introduced during which the Ghana Health Service (GHS) recorded coverage rates through routine data collection. Transmission studies conducted in the Upper Denkyira East Municipal (UDEM) of the forest zone of Ghana in 2002 and 2006 had shown that annual treatments with ivermectin had hardly any effect on the transmission of Onchocerca volvulus by the vector Simulium sanctipauli. In order to establish whether or not this was due to an insufficient compliance to the CDTI programme, an additional questionnaire survey was carried out in 2013 following those conducted in 2002 and 2006. The repeat transmission survey conducted in 2013 in the same area revealed that the vector S. sanctipauli had apparently disappeared from the rivers Ofin and Pra due to gold mining activities. In 2006 and 2013, we conducted surveys using structured questionnaires to address issues related to compliance and to compare results on the effectiveness of CDTI. A total of 692 individuals from 7 villages and 447 individuals from 9 villages were interviewed in 2006 and 2013 respectively. Questions asked included whether or not they had taken the ivermectin and reasons for not doing so when that was the case. Results were compared with the previous investigations conducted in 2002. Whereas official reported coverage rates ranged from 59 to 85% in 2006 and from 88 to 97% in 2013, compliance rates decreased from 36% in 2006 to 21% in 2013. Factors affecting compliance included fear of unpleasant side effects (pruritus and oedema), which decreased from 36% to 21% for the same period. Lack of awareness of CDTI sharply increased from 12% to 46% for the same period. Participants believed that treatments were no longer necessary due to the absence of vectors observed in 2013. There seems to be a considerable difference between coverage and compliance rates in the study communities. The difference can be attributed to the performance of the Community-Directed Distributors (CDDs) and the absence of the vector population observed in 2013. Discussions with CDDs suggested that factors that led to non-compliance were mostly side effects, unawareness of the disease by immigrants and lack of financial motivation for the CDDs. Also included was the fact that they needed to complete distribution of the drugs in the entire village, covering all households within just one week irrespective of the size of the catchment area. This, they thought was too much work for a short period of time. We propose to intensify the training of CDDs by the national Neglected Tropical Diseases Programme (NTDP) and to include the Community-based Health and Planning Services (CHPS) concept into onchocerciasis control efforts for awareness creation while the vector population and the transmission should be further monitored. The population should be made aware that the side effects they experienced from previous treatments or had heard about had reduced significantly. They also should be in the known that vector flies may return and so the risk of transmission remains.

Palp ratio as a field identification tool for two members of the Anopheles gambiae complex in Ghana (A. melas and A. gambiae).

BACKGROUND: The Anopheles gambiae Giles complex is the most widely studied and the most important insect vector group. We explored the use of the palp ratio method as a field tool to identify A. melas and A. gambiae in Ghana. METHODS: Human landing catches were conducted to collect mosquitoes in the coastal area of Western Region of Ghana. Palps were removed and segments 3 and 4 + 5 measured using a compound microscope. DNA extraction and downstream PCR for species identification was carried out using the legs and wings. Known A. gambiae collected from the Ashanti Region of Ghana were used for comparison. RESULTS: A total of 2120 A. gambiae were collected. Lengths of segments 3 and 4 + 5 were significantly correlated in samples from both regions. Using a palp ratio of 0.81 as the cut-off value, 14.9% outliers (≥0.81) from our study area were confirmed by PCR as A. melas. PCR also confirmed outliers from the Ashanti Region with palp ratio < 0.81 (10.2%) as A. gambiae. CONCLUSION: The palp ratio method proved to be a useful tool to identify populations of salt and freshwater A. melas and A. gambiae.

Does irrigated urban agriculture influence the transmission of malaria in the city of Kumasi, Ghana?

To verify the possible impact of irrigated urban agriculture on malaria transmission in cities, we studied entomological parameters, self-reported malaria episodes, and household-level data in the city of Kumasi, Ghana. A comparison was made between city locations without irrigated agriculture, city locations with irrigated urban vegetable production, and peri-urban (PU) locations with rain-fed agriculture. In the rainy as well as dry seasons, larvae of Anopheles spp. were identified in the irrigation systems of the urban farms. Night catches revealed significantly higher adult anopheline densities in peri-urban and urban agricultural locations compared to non-agricultural urban locations. Polymerase chain reaction (PCR) analysis of Anopheles gambiae sensu lato revealed that all specimens processed were A. gambiae sensu stricto. The pattern observed in the night catches was consistent with household interviews because significantly more episodes of malaria and subsequent days lost due to illness were reported in peri-urban and urban agricultural locations than in non-agricultural urban locations. In Kumasi, urban agriculture is mainly practised in inland valleys, which might naturally produce more mosquitoes. Therefore more detailed studies, also in other cities with different water sources and irrigation systems, and a better spatial distribution of sites with and without urban agriculture than in Kumasi are needed.

Malaria transmission intensity and dynamics of clinical malaria incidence in a mountainous forest region of Ghana.

Background: Malaria transmission is heterogeneous. Villages close to each other may have very different transmission characteristics. The presence and abundance of malaria vectors is governed by local ecology and microclimate. Knowledge of the dynamics of transmission is important for planning and evaluation of malaria control strategies. This study investigated the heterogeneity of malaria transmission in preparation for a vaccine trial and offers insights into dynamics of malaria incidence in the forest zone of Ghana. Methods: Malaria transmission was assessed in four villages with different micro-ecological features in the forest zone of the Akwapim-Mampong Range in Ghana, water shed with rivers flowing north to Lake Volta in the south. Human landing catches (HLC) of mosquitoes were conducted and Plasmodium falciparum circumsporozoite rates were assessed by ELISA. Sporozoite prevalence, annual biting rates (ABR) and entomological inoculation rates (EIR) from the four study sites were compared with climatological and ecological data. Regression analysis was used to compare transmission data and blood parasite prevalence, parasite density (PD) and malaria episodes from children in the study area. Additionally we examined trends in confirmed clinical malaria incidence from 2005 -2012. Results: In total 1307 Anopheles gambiae s.l. and 54 An. funestus females were caught by HLC from November 2003 to August 2005. Sporozoites in Anopheles vectors in four villages ranged from 4.0 to 10.2%, ABR from 371 to 1890 and EIR from 40 to 158. Linear regression on parasitological and clinical data of children from the villages revealed that the ABR significantly influenced the parasite density (PD) of P. falciparum. Conclusion: Malaria transmission was intense and heterogeneous and corresponded to the micro-ecological differences. Malaria transmission in the early evening hours before people went to sleep was enough to sustain stable malaria. Scaling up preventive measures to reduce exposure to vectors will be effective in reducing parasitemia in children. Variations in transmission intensity must be considered when evaluating impact of control strategies and interventions such as the vaccine trials.

Use of Bacillus thuringiensis var israelensis as a viable option in an Integrated Malaria Vector Control Programme in the Kumasi Metropolis, Ghana.

BACKGROUND: Integrated Vector Control (IVC) remains the approach for managing the malaria-causing vector. The study investigated the contribution of Bacillus thuringiensis israelensis (Bti) in the control of malaria by targeting the larvae and also mapped and documented major breeding sites in the Kumasi metropolis, Ghana. METHODS: Using a hand held GPS receiver unit, major breeding sites within the metropolis were mapped out during the larval survey. Mosquito larvae were then collected from the breeding sites and reared in an insectary to obtain an F1 generation for laboratory bioassays. The minimum effective dosage of Bti Water Dispersible Granular (WDG) formulation was determined by a series of bioassays. Based on the results obtained in the laboratory, the optimum effective dosage of Bti formulations against naturally occurring larvae of the indigenous mosquito species was determined through open field trials. RESULTS: A total of 33 breeding sites were identified and geo-referenced during the larval surveys with the majority of the breeding sites located in the Asokwa sub-metropolis, Kumasi, Ghana. A Bti (3,000 International Toxic Unit (ITU)/mg) concentration of 0.026 mg/l resulted in 50% mortality whilst a concentration of 0.136 mg/l resulted in 95% mortality. Results from the open field trials with Bti showed that a dosage of 0.2 kg/ha is as effective as 0.4 kg/ha in suppressing late instars and resulting pupae. CONCLUSION: This study reveals that Bti at a very low dosage of 0.2 kg/ha is highly effective against Anopheles larvae and therefore offers viable options for the management of vector mosquitoes. Further research is needed to extend this to the field in order to determine its ability to reduce malaria incidence.

The ecology of mosquitoes in an irrigated vegetable farm in Kumasi, Ghana: abundance, productivity and survivorship.

BACKGROUND: Irrigated vegetable farms within the city of Kumasi, Ghana, create hotspots for the breeding of malaria vectors, which could lead to high transmission of malaria. This study investigated the abundance and productivity of mosquitoes in an irrigated vegetable farm in Kumasi, Ghana. METHODS: Adult mosquito productivity was estimated five days in a week in different irrigated scheme types (dug-out wells, furrows and footprints) for 12 weeks using emergence traps. Larval sampling was done five days a week to estimate the abundance of larvae from the different irrigated schemes types. RESULTS: Mosquito breeding in the irrigated vegetable field was confined to dug-out wells, furrows and human footprints. Mosquito productivity (m2/week) was highest in the dugout wells followed by the human footprints and the least was in the furrows (11.23, 5.07 and 4.34 An. gambiae/m2/week). Larval abundance for the late instars (3rd, 4th and pupae) also followed the same trend, with the dug-out wells having the highest larval abundance followed by the human footprints and then the furrows (13.24, 6.81, 5.87 larvae/week). Mosquito productivity and abundance was negatively correlated with rainfall (R2 = 0.209; P< 0.01). CONCLUSION: This study showed that adult and larval mosquito abundance and larval survival were high in the irrigated fields in the irrigated vegetable farm. This therefore, contributed significantly to adult mosquito populations and hence malaria transmission in the city.

The effect of deltamethrin-treated net fencing around cattle enclosures on outdoor-biting mosquitoes in Kumasi, Ghana.

Classic vector control strategies target mosquitoes indoors as the main transmitters of malaria are indoor-biting and -resting mosquitoes. However, the intensive use of insecticide-treated bed-nets (ITNs) and indoor residual spraying have put selective pressure on mosquitoes to adapt in order to obtain human blood meals. Thus, early-evening and outdoor vector activity is becoming an increasing concern. This study assessed the effect of a deltamethrin-treated net (100 mg/m(2)) attached to a one-meter high fence around outdoor cattle enclosures on the number of mosquitoes landing on humans. Mosquitoes were collected from four cattle enclosures: Pen A - with cattle and no net; B - with cattle and protected by an untreated net; C - with cattle and protected by a deltamethrin-treated net; D - no cattle and no net. A total of 3217 culicines and 1017 anophelines were collected, of which 388 were Anopheles gambiae and 629 An. ziemanni. In the absence of cattle nearly 3 times more An. gambiae (p<0.0001) landed on humans. The deltamethrin-treated net significantly reduced (nearly three-fold, p<0.0001) culicine landings inside enclosures. The sporozoite rate of the zoophilic An. ziemanni, known to be a secondary malaria vector, was as high as that of the most competent vector An. gambiae; raising the potential of zoophilic species as secondary malaria vectors. After deployment of the ITNs a deltamethrin persistence of 9 months was observed despite exposure to African weather conditions. The outdoor use of ITNs resulted in a significant reduction of host-seeking culicines inside enclosures. Further studies investigating the effectiveness and spatial repellence of ITNs around other outdoor sites, such as bars and cooking areas, as well as their direct effect on vector-borne disease transmission are needed to evaluate its potential as an appropriate outdoor vector control tool for rural Africa.

Bats host major mammalian paramyxoviruses.

The large virus family Paramyxoviridae includes some of the most significant human and livestock viruses, such as measles-, distemper-, mumps-, parainfluenza-, Newcastle disease-, respiratory syncytial virus and metapneumoviruses. Here we identify an estimated 66 new paramyxoviruses in a worldwide sample of 119 bat and rodent species (9,278 individuals). Major discoveries include evidence of an origin of Hendra- and Nipah virus in Africa, identification of a bat virus conspecific with the human mumps virus, detection of close relatives of respiratory syncytial virus, mouse pneumonia- and canine distemper virus in bats, as well as direct evidence of Sendai virus in rodents. Phylogenetic reconstruction of host associations suggests a predominance of host switches from bats to other mammals and birds. Hypothesis tests in a maximum likelihood framework permit the phylogenetic placement of bats as tentative hosts at ancestral nodes to both the major Paramyxoviridae subfamilies (Paramyxovirinae and Pneumovirinae). Future attempts to predict the emergence of novel paramyxoviruses in humans and livestock will have to rely fundamentally on these data.

Henipavirus RNA in African bats.

BACKGROUND: Henipaviruses (Hendra and Nipah virus) are highly pathogenic members of the family Paramyxoviridae. Fruit-eating bats of the Pteropus genus have been suggested as their natural reservoir. Human Henipavirus infections have been reported in a region extending from Australia via Malaysia into Bangladesh, compatible with the geographic range of Pteropus. These bats do not occur in continental Africa, but a whole range of other fruit bats is encountered. One of the most abundant is Eidolon helvum, the African Straw-coloured fruit bat. METHODOLOGY/PRINCIPAL FINDINGS: Feces from E. helvum roosting in an urban setting in Kumasi/Ghana were tested for Henipavirus RNA. Sequences of three novel viruses in phylogenetic relationship to known Henipaviruses were detected. Virus RNA concentrations in feces were low. CONCLUSIONS/SIGNIFICANCE: The finding of novel putative Henipaviruses outside Australia and Asia contributes a significant extension of the region of potential endemicity of one of the most pathogenic virus genera known in humans.

Doxycycline as a novel strategy against bancroftian filariasis-depletion of Wolbachia endosymbionts from Wuchereria bancrofti and stop of microfilaria production.

Chemotherapy of onchocerciasis by doxycycline, which targets symbiotic Wolbachia endobacteria, has been shown to result in a long-term sterility of adult female worms and corresponding absence of microfilariae. It represents an additional chemotherapeutic approach. The aim of this study was to determine whether a similar regimen would also show efficacy against Wuchereria bancrofti. Ghanaian individuals ( n=93) with lymphatic filariasis and a minimum microfilaremia of 40 microfilariae/ml were included in a treatment study consisting of four arms: (1) doxycycline 200 mg/day for 6 weeks; (2) doxycycline as in (1), followed by a single dose of ivermectin after 4 months; (3) ivermectin only; or (4) no treatment during observation period of 1 year (ivermectin at the end of the study). Doxycycline treatment resulted in a 96% loss of Wolbachia, as determined by real time PCR from microfilariae. After 12 months, doxycycline had led to a 99% reduction of microfilaremia when given alone, and to a complete amicrofilaremia together with ivermectin. In contrast, after ivermectin treatment alone a significant presence of microfilariae remained (9% compared to pretreatment), as known from other studies. This study shows that doxycycline is also effective in depleting Wolbachia from W. bancrofti. It is likely that the mechanism of doxycycline is similar to that in other filarial species, i.e., a predominant blockade of embryogenesis, leading to a decline of microfilariae according to their half-life. This could render doxycycline treatment an additional tool for the treatment of microfilaria-associated diseases in bancroftian filariasis, such as tropical pulmonary eosinophilia and microfiluria.