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Prior to 1999, Ghana was not known to be a Cutaneous Leishmaniasis (CL) endemic country. The outbreak of CL in the Volta Region in 1999 resulted in the need for urgent investigations to understand the disease transmission dynamics. Monthly entomological collections from January 2006 to May 2007 revealed 99.8% (21,980/22,016) of sand flies collected were Sergentomyia and 0.2% (36/22,016) Phlebotomus, the putative vector of CL in the Old World. The density of Phlebotomus was so low, their vectorial role is uncertain. This study was therefore aimed at determining the possible vector species in the outbreak area. We collected sand flies from homes to examine anthropophily, by testing their blood meals. The blood-meal sources of 275 (20.5%) of 1342 collected female sand flies revealed the highest proportion of animal blood source was chicken (33.8%) and goat (12.4%). Humans accounted for 32% of blood meal sources. Identified species included: Sergentomyia africana africana (26.9%), S. ingrami (34.5%) and S. simillima (38.2%), which were found to have single and mixed blood-meals, with S. simillima having the highest proportion that contained human blood (14.6%). Using molecular methods, we examined unfed flies and flies found anthropophilic for the presence of Leishmania major DNA. Poolscreening analysis revealed 3 pools of S. ingrami positive with L. major DNA, with an infection rate of 1.27% (95% confidence interval: 2.467–3.647). This study showed that some Sergentomyia species are anthropophilic, contain L. major DNA, and could be involved in the transmission of CL in Ghana. There is however the need to conclusively validate the role of S. ingrami as a vector of L. major in Ghana, by isolating the parasite and initiating experimental transmission studies.

The scale up of indoor residual spraying (IRS) and insecticide treated nets have contributed significantly to global reductions in malaria prevalence over the last two decades. However, widespread pyrethroid resistance has necessitated the use of new and more expensive insecticides for IRS. Partial IRS with pirimiphos-methyl in experimental huts and houses in a village-wide trial was evaluated against Anopheles gambiae s.l. in northern Ghana. Four different scenarios in which either only the top or bottom half of the walls of experimental huts were sprayed, with or without also spraying the ceiling were compared. Mortality of An. gambiae s.l. on partially sprayed walls was compared with the standard procedures in which all walls and ceiling surfaces are sprayed. A small-scale trial was then conducted to assess the effectiveness, feasibility, and cost of spraying only the upper walls and ceiling as compared to full IRS and no spraying in northern Ghana. Human landing catches were conducted to estimate entomological indices and determine the effectiveness of partial IRS. An established transmission dynamics model was parameterized by an analysis of the experimental hut data and used to predict the epidemiological impact and cost effectiveness of partial IRS for malaria control in northern Ghana. In the experimental huts, partial IRS of the top (IRR 0.89, p = 0.13) or bottom (IRR 0.90, p = 0.15) half of walls and the ceiling was not significantly less effective than full IRS in terms of mosquito mortality. In the village trial, the annual entomological inoculation rate was higher for the unsprayed control (217 infective bites/person/year (ib/p/yr)) compared with the fully and partially sprayed sites, with 28 and 38 ib/p/yr, respectively. The transmission model predicts that the efficacy of partial IRS against all-age prevalence of malaria after six months would be broadly equivalent to a full IRS campaign in which 40% reduction is expected relative to no spray campaign. At scale, partial IRS in northern Ghana would have resulted in a 33% cost savings ($496,426) that would enable spraying of 36,000 additional rooms. These findings suggest that partial IRS is an effective, feasible, and cost saving approach to IRS that could be adopted to sustain and expand implementation of this key malaria control intervention.

Background: Significant decrease in malaria morbidity and mortality have been attained using long-lasting insecticide treated nets and indoor residual spraying. Selective pressure from these control methods influences change in vector bionomics and behavioural pattern. There is a need to understand how insecticide resistance drives behavioural changes within vector species. This study aimed to determine the spatio-temporal dynamics and biting behaviour of malaria vectors in different ecological zones in Ghana in an era of high insecticide use for public health vector control.Methods: Adult mosquitoes were collected during the dry and the rainy seasons in 2017 and 2018 from five study sites in Ghana in different ecological zones. Indoor and outdoor biting mosquitoes were collected per hour from 18:00 to 06:00 hours employing the human landing catches (HLC) technique. Morphological and molecular species identification of vectors were done using identification keys and PCR respectively. Genotyping of insecticide resistant markers was done using the TaqMan SNP genotyping probe-based assays. Detection of P. falciparum sporozoites was determined using PCR.Results: A total of 50,322 mosquitoes belonging to four different genera were collected from all the study sites during the sampling seasons in 2017 and 2018. Among the Anophelines were Anopheles gambiae s.l. 93.16%, (31055/33,334), An. funestus 2.07%, (690/33,334), An. pharoensis 4.63%, (1545/33,334), and An. rufipes 0.13% (44/33,334). Overall, 76.40%, (25,468/33,334) of Anopheles were collected in the rainy season and 23.60%, (7,866/33,334) in the dry season. There was a significant difference (z = 2.410; p = 0.0160) between indoor biting (51.09%; 15,866/31,055) and outdoor biting An. gambiae s.l. (48.91%; 15,189/31,055). The frequency of the Vgsc-1014F mutation was slightly higher in indoor biting mosquitoes (54.90%) than outdoors (45.10%). Overall, forty-four pools (44) of samples were positive for P. falciparum CSP giving an overall sporozoite rate of 0.07%.Conclusion: Anopheles gambiae s.l. were more abundant indoors across all ecological zones of Ghana. The frequency of G119S was higher in indoor than outdoor from all the study sites, but higher sporozoite rates in outdoor mosquitoes in Dodowa and Kpalsogu. There is thus, an urgent need for a supplementary malaria control intervention to control outdoor biting mosquitoes.

Many lymphatic filariasis (LF) endemic countries, including Ghana, have successfully implemented mass drug administration (MDA) and made significant progress towards the elimination of the disease as a public health problem. Unfortunately, the existence of individuals who seldom or never take part in MDA pose a threat to this success, as they may serve as reservoirs of infection, re-infecting their communities. In this study we implemented strategies to identify and treat these individuals, while also assessing their level of infection, to inform programme actions. The study was undertaken in the Ahanta West hotspot district in Ghana, which has received more than 17 rounds of MDA. Through the community registers used in recording participation in MDAs, we identified and offered treatment to individuals who were ineligible or inadvertently missed the last MDA in April 2021 (Engage and Treat – E&T), or testing using the filariasis test strip followed by treatment to community members who for various reasons chose not to participate in the last MDA (Test and Treat – T&T). During the study, 23,879 individuals ranging from 5 to 98 years were reached, of whom 78% were not captured in the MDA register. Among the E&T group, 75.06% willingly received and swallowed the treatment drugs. The remaining 24.94% were offered testing followed by a re-engagement to receive the drug in the T&T group. Overall, 22,830 (95.61%) of participants were treated by either strategy. Of the participants in the T&T group, 516 (8.66%; 95% CI= 7.96 – 9.41) were positive by the FTS. The highest antigen prevalence was detected among children 5 to 10 years, with 16.59% (95% CI= 12.02 – 22.06) and 22.54% (95% CI= 17.11 – 28.74) among females and males, respectively. Mapping of the data revealed that most infections are in a few select communities. Of the 516 FTS positives, 27.33% reportedly missed MDA once, 18.41% missed MDA twice and 54.26% missed all of the last three MDAs. The main reasons for missing MDA included absence (25.49%), travel (21.24%), being unaware of MDA (20.27%), refusals to take the drug (10.65%), illnesses (7.07%) and fear of adverse events (6.13%). This study demonstrates that greater sensitization and engagement strategies, with a test and treat strategy reserved for the most hesitant individuals, could significantly increase the number of individuals who receive treatment and therefore help districts reach their elimination targets by reducing the remaining reservoir or infection. NTD programmes require new tools to help them identify, engage and treat these individuals, as part of their overall monitoring and evaluation strategy.

Background: The presence of breeding sites and distribution of species of Simulium damnosum s.l are critical in understanding the epidemiology of onchocerciasis and evaluating the impact of elimination interventions. Reports on breeding sites and species distribution of members of S. damnosum s.l in Cameroon are scarce and the few available date back to more than 3 decades. The aim of this study is to provide information on S. damnosum breeding sites across the rainy (RS) and dry (DS) seasons and the species composition in three different regions in Cameroon: Southwest (SW), Northwest (NW) and North (N).Methods: A cross-sectional two-seasons study was carried out in three regions with different ecological characteristics (SW – rainforest; NW- mixed forest-Guinea savanna; N- Sudan savanna). Pre-control onchocerciasis endemicity, relief maps and historical entomological information were used to identify potential rivers for purposive sampling. Sampled larvae were fixed in Carnoy’s solution, sorted and S. damnosum s.l larvae stored until identification by cytotaxonomy. Geographical coordinates of potential breeding sites were recorded to produce maps using ArcGIS, while SPSS used for Chi-square test to estimate difference between blackfly seasonal breeding rates.Results: 237 potential breeding sites were sampled (RS=81; DS=156) and 72 were found positive for S. damnosum s.l. SW had the most positive sites [67 (RS=24; DS=43)], having significant difference in rate of breeding between the seasons (pSimulium damnosum s.s and S. sirbanum were the main species in N while S. squamosum and S. mengense the predominant species in NW and SW. S. soubrense and S. yahense were uniquely recorded in SW.Conclusions: A comprehensive mapping of breeding sites requires rainy and dry seasons sampling. Study also demonstrates S. damnosum s.l breeding site survey is achievable in forest as in savanna zones. Not all potential breeding sites are actual breeding sites. Observation of S. soubrense in SW indicates changes in species composition over time and could affect onchocerciasis epidemiology in this area.

Background The presence of breeding sites and distribution of species of Simulium damnosum sensu lato are critical in understanding the epidemiology of onchocerciasis and evaluating the impact of elimination interventions. Reports on breeding sites and species distribution of members of S. damnosum s.l. in Cameroon are scarce and the few ones available date back to more than three decades. The aim of this study is to provide information on S. damnosum breeding sites across the rainy (RS) and dry (DS) seasons and the species composition in three different regions in Cameroon: Southwest (SW), Northwest (NW) and North (N). Methods A cross-sectional two-season study was carried out in three regions with different ecological characteristics (SW—rainforest; NW—mixed forest–Guinea savanna; N—Sudan savanna). Pre-control onchocerciasis endemicity, relief maps and historical entomological information were used to identify potential rivers for purposive sampling. Sampled larvae were fixed in Carnoy’s solution and sorted, and S. damnosum s.l. larvae were stored until identification by cytotaxonomy. Geographical coordinates of potential breeding sites were recorded to produce maps using ArcGIS, while Chi-square tests in SPSS were used to test for any differences between black fly seasonal breeding rates. Results A total of 237 potential breeding sites were sampled (RS = 81; DS = 156) and 72 were found positive for S. damnosum s.l. The SW had the most positive sites [67 (RS = 24; DS = 43)], with a significant difference in the rate of breeding between the seasons (P Simulium damnosum sensu stricto and S. sirbanum were the main species in the N, while S. squamosum and S. mengense were the predominant species in the NW and SW. Simulium soubrense and S. yahense were uniquely recorded in the SW. Conclusions A comprehensive mapping of breeding sites requires rainy and dry seasons sampling. This study demonstrates that a breeding site survey of S. damnosum s.l. is achievable in forest as well as savanna zones. Not all potential breeding sites are actual breeding sites. Observation of S. soubrense in the SW indicates changes in species composition over time and could affect onchocerciasis epidemiology in this area. Graphical abstract

Onchocerciasis and lymphatic filariasis are targeted for elimination, primarily using mass drug administration at the country and community levels. Elimination of transmission is the onchocerciasis target and global elimination as a public health problem is the end point for lymphatic filariasis. Where program duration, treatment coverage, and compliance are sufficiently high, elimination is achievable for both parasites within defined geographic areas. However, transmission has re-emerged after apparent elimination in some areas, and in others has continued despite years of mass drug treatment. A critical question is whether this re-emergence and/or persistence of transmission is due to persistence of local parasites—i.e., the result of insufficient duration or drug coverage, poor parasite response to the drugs, or inadequate methods of assessment and/or criteria for determining when to stop treatment—or due to re-introduction of parasites via human or vector movement from another endemic area. We review recent genetics-based research exploring these questions in Onchocerca volvulus, the filarial nematode that causes onchocerciasis, and Wuchereria bancrofti, the major pathogen for lymphatic filariasis. We focus in particular on the combination of genomic epidemiology and genome-wide associations to delineate transmission zones and distinguish between local and introduced parasites as the source of resurgence or continuing transmission, and to identify genetic markers associated with parasite response to chemotherapy. Our ultimate goal is to assist elimination efforts by developing easy-to-use tools that incorporate genetic information about transmission and drug response for more effective mass drug distribution, surveillance strategies, and decisions on when to stop interventions to improve sustainability of elimination.

Aedes aegypti (L.) (Diptera: Culicidae) is a diurnal feeder that lives in close association with human populations. It is the principal vector of yellow fever, dengue fever and the Zika Virus. Issues of arboviral diseases have been on the ascendency in most countries including Ghana where Aedes mosquito is the main vector of yellow fever. A comparative study of the biting behavior of Ae. aegypti and the identification of subspecies were undertaken using molecular technique. Standard human landing technique was used to collect both indoor and outdoor biting mosquitoes at three zones located in the Upper East (Bolgatanga), Upper West (Nadowli), and Northern (Damongo) Regions of Ghana during the dry and rainy seasons between 0600 and 1800 Greenwich Mean Time (GMT). All collected mosquitoes were identified morphologically using taxonomic keys. random amplified polymorphic DNA polymerase chain reaction was used to categorize Ae. aegypti into subspecies. Adult female Aedes mosquitoes identified formed 62% (n = 1,206) of all female mosquitoes collected. Aedes aegypti 98% and Aedes vittatus 2% were the only Aedes species identified. Bolgatanga recorded the largest number of Ae. aegypti 42%, whereas Nadowli 22% recorded the least. Aedes vittatus was observed in Nadowli. Aedes aegypti exhibited a bimodal biting behavior peaking at 0600–0800 GMT and 1500–1600 h GMT. Molecular findings revealed 69% Ae. aegypti aegypti and 31% Ae. aegypti formosus as the two subspecies (n = 110). This information is important for implementing effective vector control programs in the three regions of the northern Ghana.

Background Malaria is transmitted through the bite of Plasmodium‐infected adult female Anopheles mosquitoes. Ivermectin, an anti‐parasitic drug, acts by killing mosquitoes that are exposed to the drug while feeding on the blood of people (known as blood feeds) who have ingested the drug. This effect on mosquitoes has been demonstrated by individual randomized trials. This effect has generated interest in using ivermectin as a tool for malaria control. Objectives To assess the effect of community administration of ivermectin on malaria transmission. Search methods We searched the Cochrane Infectious Diseases Group (CIDG) Specialized Register, CENTRAL, MEDLINE, Embase, LILACS, Science Citation index ‐ expanded, the World Health Organization (WHO) International Clinical Trials Registry Platform, ClinicalTrials.gov, and the National Institutes of Health (NIH) RePORTER database to 14 January 2021. We checked the reference lists of included studies for other potentially relevant studies, and contacted researchers working in the field for unpublished and ongoing trials. Selection criteria We included cluster‐randomized controlled trials (cRCTs) that compared ivermectin, as single or multiple doses, with a control treatment or placebo given to populations living in malaria‐endemic areas, in the context of mass drug administration. Primary outcomes were prevalence of malaria parasite infection and incidence of clinical malaria in the community. Data collection and analysis Two review authors independently extracted data on the number of events and the number of participants in each trial arm at the time of assessment. For rate data, we noted the total time at risk in each trial arm. To assess risk of bias, we used Cochrane's RoB 2 tool for cRCTs. We documented the method of data analysis, any adjustments for clustering or other covariates, and recorded the estimate of the intra‐cluster correlation (ICC) coefficient. We re‐analysed the trial data provided by the trial authors to adjust for cluster effects. We used a Poisson mixed‐effect model with small sample size correction, and a cluster‐level analysis using the linear weighted model to adequately adjust for clustering. Main results We included one cRCT and identified six ongoing trials. The included cRCT examined the incidence of malaria in eight villages in Burkina Faso, randomized to two arms. Both trial arms received a single dose of ivermectin 150 µg/kg to 200 µg/kg, together with a dose of albendazole. The villages in the intervention arm received an additional five doses of ivermectin, once every three weeks. Children were enrolled into an active cohort, in which they were repeatedly screened for malaria infection. The primary outcome was the cumulative incidence of uncomplicated malaria in a cohort of children aged five years and younger, over the 18‐week study. We judged the study to be at high risk of bias, as the analysis did not account for clustering or correlation between participants in the same village. The study did not demonstrate an effect of Ivermectin on the cumulative incidence of uncomplicated malaria in the cohort of children over the 18‐week study (risk ratio 0.86, 95% confidence interval (CI) 0.62 to 1.17; P = 0.2607; very low‐certainty evidence). Authors' conclusions We are uncertain whether community administration of ivermectin has an effect on malaria transmission, based on one trial published to date., Plain language summary Malaria control using ivermectin What is the aim of this review? The aim of this Cochrane Review was to find out if giving the drug ivermectin to entire communities could reduce malaria transmission. We examined all relevant studies to answer this question, and found one relevant study. Key messages It is not possible to say at this point if treating an entire community with ivermectin reduces malaria. Several research studies are in progress; we anticipate they will provide more answers in the future. What was studied in the review? Malaria is a disease transmitted to humans through the bite of mosquitoes infected with Plasmodium parasites. It results in nearly half a million deaths every year. Ivermectin is a drug that is given to whole communities to control the parasites that are responsible for elephantiasis and river blindness. It has been observed that ivermectin can kill mosquitoes when they feed on the blood of people who have taken this medication. Therefore, it is believed that by giving this drug to whole communities, it will kill many mosquitoes, and could reduce malaria transmission. In this review, we assessed whether treating entire communities with ivermectin would reduce malaria transmission. We looked for studies from different sources, and only included studies that took place in communities with malaria, and that randomly assigned groups of people to ivermectin or a control, which could be a placebo or standard community drug treatments. We wanted to know if the treatment influenced the occurrence of malaria in the community. What are the main results of the review? One study met the inclusion criteria. This study included eight villages in Burkina Faso, which were randomly assigned to receive ivermectin or a control. All villages received ivermectin, as part of the scheduled control of lymphatic filariasis. In addition, the treatment villages received five more doses of ivermectin, once every three weeks. The effect of ivermectin on malaria was measured in children younger than five years of age. In these children, the treatment did not show a notable difference in the presence of malaria between the treatment and control groups (very low‐certainty evidence). Therefore, it is not possible to say at this point if the treatment of entire communities with ivermectin has an effect on reducing malaria. Several studies are currently ongoing; we anticipate they will provide more answers in the future. How up‐to‐date is this review? We searched for studies published up to 14 January 2021.

Background Onchocerciasis is caused by a nematode worm Onchocerca volvulus, which is transmitted in Sudan by black fly vectors of the Simulium damnosum sensu lato species complex. In Sudan, the disease is found in four foci where fast flowing rivers provide suitable breeding sites for the Simulium vector flies. The construction of dams and irrigation schemes for agricultural purposes has affected black fly breeding and distribution, such as in Merowe Dam in Abu-Hamed focus, where the perennially flowing water downstream of the Dam created new vector breeding sites, thereby, changing the pattern of disease transmission and creating public health problems. Based on this situation, this study was carried out to measure the effect of the Upper Atbara and Setit Dam complex on the distribution of Simulium damnosum s.l. breeding sites and on disease elimination in the Galabat sub-focus in eastern Sudan. Methods Aquatic stages of Simulium were collected between October and November 2009, prior to the construction of the dam complex, and again in 2013 and 2015 while the dam complex construction was ongoing. Results A total of 40 breeding sites were identified at the beginning of the study. After the construction of the dam complex in 2015, seventeen previously mapped breeding sites were inaccessible as they had been flooded by the dam complex’s lake when reach its maximum size. Three species were obtained from different locations: S. damnosum s.l., S. griseicolle, and S. adersi. Conclusions This study has shown a link between the construction of the dam complex and a reduction in the breeding sites of black fly vectors. This reduction has limited the Galabat sub-focus to a small area at the upper Atbara River which become the end of the focus. To sustain the success achieved in onchocerciasis control in the Galabat sub-focus, disease control and its vector control should be strengthened in the area cross-boarding Sudan and Ethiopia. Electronic supplementary material The online version of this article (10.1186/s12879-019-4113-1) contains supplementary material, which is available to authorized users.

The aim of this paper is twofold. Firstly, to present design research work that crosses into specialist medical areas, in this instance that of antimicrobial resistance (AMR); and secondly, to contribute to the debate on the need for design driven research to explore alternative ways of delivering healthcare by focusing on preventive design responses. These points are addressed in the exploration of an international design-led research project, Dust Bunny. The project uniquely combines design research and microbiology to provide an informed assessment of societal practices in domestic cleanliness and to co-create novel solutions that reduce infections in the home. The findings and experiences from this unique transdisciplinary collaboration (design, microbiology, epidemiology and social science) reveal several insights on the challenges and opportunities on challenging the boundaries of design thinking.

In response to the developing global health crisis arising from the diminishing efficacy of antibiotics, a design-led collaboration between academics from the UK and Ghana explored hygiene practices across different home environments in Ghana. In doing so, we developed our understanding of the home as a source of infection stemming from bacteria, some of which are disease-causing pathogens carried by dust that could carry antimicrobial resistance. These insights were then used by householders to co-design context-specific cleaning practices that sought to mitigate the impacts of pathogen exposure and antimicrobial resistance. This case study outlines the Dust Bunny project and draws on reflections and interviews with the wider research team to offer insights into the challenges and opportunities for design research exemplified in this project, which may be applicable to other Global South-Global North project collaborations.

Aedes-borne viral diseases mainly Yellow Fever (YF), Dengue (DEN), Zika (ZIK) and Chikungunya (CHK) have contributed to many deaths’ in the world especially in Africa. There have been major outbreaks of these diseases in West Africa. Although, YF outbreaks have occurred in Ghana over the years, no outbreak of DEN, ZIK and CHK has been recorded. However, the risk of outbreak is high due to its proximity to West African countries where outbreaks have been recently been recorded. This study surveyed the mosquito fauna to assess the risk of transmission of Yellow fever (YFV), Dengue (DENV), Chikungunya (CHKV) and Zika (ZIKV) viruses in Larabanga and Mole Game Reserve areas in Northern Ghana. The immature and adult stages ofAedesmosquitoes were collected from Larabanga and Mole Game Reserve area. There was a significant (P>0.001) number of mosquitoes collected during the rainy season than the dry season. A total of 1,930Aedesmosquitoes were collected during the rainy season and morphologically identified. Of these, 1,915 (99.22%) wereAedes aegyptiand 15 (0.22%) wereAedes vittatus. During the dry season, 27Ae.aegyptimosquitoes were collected. A total of 415Ae.aegyptimosquitoes were molecularly identified to subspecies level of whichAe.(Ae) aegypti aegyptiwas the predominant subspecies. BothAe.aegypti aegypti and Ae aegypti formosusexist in sympatry in the area. AllAedespools (75) were negative for DENV, ZIKV and CHKV when examined by RT- PCR. Three Larval indices namely House Index, HI (percentage of houses positive forAedeslarvae or pupae), Container Index, CI (the percentage of containers positive forAedeslarvae or pupae) and Breteau Index, BI (number of positive containers per 100 houses inspected) were assessed as a measure for risk of transmission in the study area. The HI, CI and BI for both sites were as follows; Mole Game Reserve (HI, 42.1%, CI, 23.5% and BI, 100 for rainy season and 0 for all indices for dry season) and Larabanga (39%, 15.5% and 61 for rainy season and 2.3%, 1.3% and 2.3 for dry season). The spatial distribution ofAedesbreeding sites in both areas indicated thatAedeslarvae were breeding in areas with close proximity to humans. Lorry tires were the main source ofAedeslarvae in all the study areas. Information about the species composition and the potential role ofAedesmosquitoes in future outbreaks of the diseases that they transmit is needed to design efficient surveillance and vector control tools.

Aedes-borne viral diseases mainly Yellow Fever (YF), Dengue (DEN), Zika (ZIK) and Chikungunya (CHK) have contributed to many deaths’ in the world especially in Africa. There have been major outbreaks of these diseases in West Africa. YF outbreaks have occurred in Ghana. Although Ghana has not recorded any outbreak of DEN, ZIK and CHK, the risk is high due to its proximity to West African countries where outbreaks have been recently been recorded. This study assessed the risk of transmission of Yellow fever (YFV), Dengue (DENV), Chikungunya (CHKV) and Zika (ZIKV) viruses in Larabanga and Mole Game Reserve areas in Northern Ghana. The immature and adult stages of Aedes mosquitoes were collected from Larabanga and Mole Game Reserve area. There was a significant (P>0.001) number of mosquitoes collected during the rainy season than the dry season. A total of 1,930 Aedes mosquitoes were collected during the rainy season and morphologically identified. Of these, 1,915 (99.22%) were Aedes aegypti and 15 (0.22%) were Aedes vittatus. During the dry season, 27 Aedes aegypti mosquitoes were collected. A total of 415 Ae. aegypti mosquitoes were molecularly identified to subspecies level of which Aedes (Ae) aegypti aegypti was the predominant subspecies. Both Ae. aegypti aegypti and Ae aegypti formosus exist in sympatry in the area. All Aedes pools (75) were negative for DENV, ZIKV and CHKV when examined by RT-PCR. Three Larval indices namely House Index, HI (percentage of houses positive for Aedes larvae or pupae), Container Index, CI (the percentage of containers positive for Aedes larvae or pupae) and Breteau Index, BI (the number of positive containers (with larvae and/or pupae per 100 inspected houses) were assessed as a measure for risk of transmission. The HI, CI and BI for both sites were as follows; Mole game reserve (HI, 42.1%, CI, 23.5% and BI, 100 for rainy season and 0 for all indices for dry season) and Larabanga (39%, 15.5% and 61 for rainy season and 2.3%, 1.3% and 2.3 for dry season). The spatial distribution of Aedes breeding sites in both areas indicated that Aedes larvae were breeding in areas with close proximity to humans. Lorry tires were the main source of Aedes larvae in all the study areas. Information about the species composition and the potential role of Aedes mosquitoes in future outbreaks of the diseases that they transmit is needed to design efficient surveillance and vector control tools.

Background Lymphatic filariasis (LF) is endemic in Ghana, and the country has implemented the GPELF strategy since 2000 with significant progress made in the control of the disease. However, after several years of mass drug administration (MDA) implementation, there is persistent transmission in 17 of the 98 endemic districts in the country. Current approaches to surveillance are clearly unable to target untreated individuals and new strategies are required to address the endgame challenges to enhance LF elimination as a public health problem in endemic countries. Community registers are used during MDAs to enumerate community members, their age, gender, house numbers, and records of their participation in MDAs. These MDA registers represent an untapped opportunity to identify and characterize non-compliance and inform appropriate programmatic actions. In this study, we analyzed the data presented in the registers to assess the coverage and individuals’ compliance in MDA. Methods The information in the MDA registers were assessed to verify the reported coverages obtained from the district. The community registers were obtained from the district health offices and the data from each individual record was entered into a database. A simple questionnaire was used to cross-check the participation of study participants in the 2017 MDA. The questionnaire solicited data on: participation in the 2017 MDA, reasons for not taking part in the MDA, adverse events experienced, what was done for the adverse events, and willingness to participate in subsequent MDAs. Results We found that 40.1% of the population in the registers missed at least one MDA in 3 years (2016–2018) and the majority of them were between 10–30 years of age. The results of the questionnaire assessment indicated that 13.8% of the respondents did not receive treatment in 2017 for various reasons, the most prominent among them being “absence/travel” (37.1%). Data in the registers were used to verify the treatment coverage for the years 2017 and 2018, and reviewed against the reported coverage obtained from the district. Significant differences between the reported and verified coverages were only observed in four communities. However, the assessment also revealed that the reported coverage was only accurate in 33.3% of cases. Conclusions The MDA registers allow for the identification of eligible individuals who were not reached during any MDA round. Thus, the MDA registers could be utilized at the community and programme levels to identify missing and untreated individuals, appropriately address their non-compliance to MDA, and thereby improve MDA coverage in each implementation unit and monitor the progress towards elimination of LF. The challenges observed through the review of the registers also offer opportunities to improve the training given to the community drug distributors., Author summary Elephantiasis is a disease that is common in Ghana. Since the year 2000 the country has implemented a control programme aimed at eliminating the disease as a public health problem, through the yearly treatment of entire communities in districts where the disease is found. Achievements have been made, and treatment has stopped in many districts; however, some districts have received many years of treatment without successfully reducing the infections below the targeted levels. Many individuals go untreated every year, as can be observed from the low reported treatment coverage after MDA. This untreated population may explain why some districts have failed to achieve the criteria for stopping treatment. Fortunately, the treatments are recorded in community registers, which contain the details of all community members, as well as their treatment participation. These registers can therefore be used to identify people who have not received treatment and design appropriate interventions to reach them. Through these registers, we were able to identify and characterize individuals who were not treated between 2016 and 2018 in 10 communities. We found that 4 out of 10 people missed at least one treatment between 2016 and 2018, and the majority of them were 10–30 years of age. Using the information from the registers we were also able to verify the proportion of people who took part in the treatments in 2017 and 2018, and compared this to what was reported. This comparison revealed that the reporting was only accurate in 33% of cases. A questionnaire was also used to cross-check the participation of study participants in the 2017 MDA, with results indicating that 13.8% of the respondents did not receive treatment for various reasons, the most prominent among them being “absence/travel” (37.1%). Although these registers are seldom used by the Lymphatic Filariasis Control Programme for anything other than recording and reporting treatment information, this study shows that the registers provide an opportunity to identify individuals who have not received treatment, develop a plan to appropriately address their reasons for not taking part in the treatments, target these untreated individuals to improve the overall proportion of people taking part in MDA, and assess the accuracy of reported coverage information. These activities will allow the programme to better monitor the progress towards, and ultimately achieve, elimination of lymphatic filariasis as a public health problem.

IntroductionUnderstanding the feeding behavior and host choice of sand flies provides valuable information on vector-host relationships and elucidates the epidemiological patterns of leishmaniasis transmission. Blood meal analysis studies are essential for estimating the efficiency of pathogen transmission, assessing the relative human disease risk, and assist in identifying the other potential hosts of leishmaniasis. In Sudan and most of East Africa, there are large remaining gaps in knowledge regarding the feeding habits of phlebotomine vectors. The study aimed to identify the blood meal sources and, therefore, the host preferences of the principal vectors Phlebotomus orientalis and Ph. papatasi in leishmaniasis endemic areas of eastern and central Sudan.Materials and methodsSand flies were collected from two endemic villages in eastern and central Sudan using CDC light traps and sticky traps. The phlebotomine sand flies were morphologically and then molecularly identified. The source of blood meal of the engorged females was determined using a multiplex PCR methodology and specific primers of cytochrome b gene of mitochondrial DNA for human, goat, cow, and dog. The detection of the Leishmania parasite was done using PCR.ResultsThe total number of collected female phlebotomine sand flies was 180. Morphological identification revealed the abundance of Ph. orientalis 103 (57.2%), Ph. papatasi 42 (23.3%), Ph. bergeroti 31 (17.2%), Ph. rodhaini 2 (1.1%) and Ph. duboscqi 2 (1.1%) in the study sites. Out of the 180 collected, 31 (17%) were blood-fed flies. Three species were blood-fed and molecularly identified: Ph. papatasi (N = 7, 22.6%), Ph. bergeroti (N = 9, 26%), and Ph. orientalis (N = 15, 48.4%). Blood meal analysis revealed human DNA in two Ph. orientalis (6.4%), hence, the anthropophilic index was 13.3%.ConclusionsMultiplex PCR protocol described here allowed the identification of blood meal sources of many vertebrate species simultaneously. The results indicate that wild-caught Ph. orientalis are anthropophilic in the study areas. Further studies on larger blood-fed sample size are required to validate the potential applications of this technique in designing, monitoring and evaluating control programs, particularly in investigating the potential non-human hosts of leishmaniasis.

Trypanosomiasis, leishmaniasis, and malaria are protozoan infections of public health importance with thousands of new cases recorded annually. Control of these infection(s) with existing chemotherapy is limited by drug toxicity, lengthy parenteral treatment, affordability, and/or the emergence of resistant strains. Medicinal plants on the other hand are used in the treatment of various infectious diseases although their chemical properties are not fully evaluated. In this study, we screened 112 crude extracts from 72 selected Ghanaian medicinal plants for anti-Trypanosoma, anti-Leishmania, and anti-Plasmodium activities in vitro and investigated their mechanisms of action. Twenty-three extracts from 20 plants showed significant antiprotozoan activity against at least 1 of 3 protozoan parasites screened with IC50 values less than 20 μg/ml. Eleven extracts showed high anti-Trypanosoma activity with Bidens pilosa whole plant and Morinda lucida leaf extracts recording the highest activities. Their IC50 (selectivity index [SI]) values were 5.51 μg/ml (35.00) and 5.96 μg/ml (13.09), respectively. Nine extracts had high anti-Leishmania activity with Annona senegalensis and Cassia alata leaf extracts as the most active. Their IC50 (SI) values were 10.8 μg/ml (1.50) and 10.1 μg/ml (0.37), respectively. Six extracts had high anti-Plasmodium activity with the leaf and stem-bark extracts of Terminalia ivorensis recording the highest activity. Their IC50 (SI) values were 7.26 μg/ml (129.36) and 17.45 μg/ml (17.17), respectively. Only M. lucida at 25 μg/ml induced significant apoptosis-like cell death in Trypanosoma parasites. Anti-Leishmania active extracts induced varying morphological changes in Leishmania parasites such as multiple nuclei and/or kinetoplast, incomplete flagella division, or nuclear fragmentation. Active extracts may be potential sources for developing new chemotherapy against these infections.

The transition from onchocerciasis control to elimination requires country programmes to rethink their approach to a variety of activities as they move from addressing morbidity to addressing transmission of the parasite. Although the 2016 WHO guidelines provide extensive recommendations, it was beyond the scope of the document to provide guidance on all aspects of the transition. This paper will discuss some of the important issues that programmes are grappling with as they transition to elimination and provide some potential approaches that programmes can use to address them. Although there are some data to support some aspects of the suggested approaches, operational research will be needed to generate data to support these approaches further and to determine how programmes could best tailor them to their own unique epidemiological challenges. Good communication between the national programmes and the broader global programme will facilitate the clear articulation of programmatic challenges and the development of the evidence to support programme decision-making.

Background Cutaneous leishmaniasis (CL) is the most common type of leishmaniasis, a neglected tropical disease caused by parasites of the genus Leishmania. In Ghana, some studies in the Volta region have detected Leishmania parasites among persons with skin ulcers. Methodology/Principal findings Using a cross-sectional study design, the prevalence of CL in three communities of the Oti Region of Ghana was investigated. Demographic and epidemiological data were obtained by a structured interviewer administered questionnaire. A total of 426 (12.4%) out of 3,440 participants screened had at least one skin ulcer. Of 595 skin ulcers sampled and tested by PCR for Leishmania infection, 150 (25.2%) ulcers from 136 individuals tested positive, accounting for an overall CL prevalence of 31.9% among persons with skin ulcers. Individual community CL prevalence of 23.2%, 29.8%, and 36.8% was observed in Ashiabre, Keri, and Sibi Hilltop respectively among persons with skin ulcers. Conclusions/Significance Confirmation of CL in the study area suggests an active cycle of transmission of Leishmania infection. The observation of skin ulcers which tested negative to Leishmania infection suggests a need to test for additional causes of skin ulcers such as Treponema pallidum pertenue and Mycobacterium ulcerans in the study area., Author summary Cutaneous leishmaniasis (CL) is a neglected tropical disease caused by parasites of the genus Leishmania and is transmitted by various species of female sandflies. CL usually begins as painless nodules and is usually characterized by skin ulcers which may be single or multiple. Although ulcers due to CL are often self-healing, they may become painful and result in scarring after healing. Typically, CL occurs on exposed parts of the body such as the neck, limbs, and face, which may be easily accessible to sandflies. Although cases of CL had previously been reported in some parts of the Volta region of Ghana, no previous case of CL had been reported in the Oti region. This study was initiated following reports of skin ulcers which were suggestive of CL in some communities of the Oti region. This study confirmed Leishmania infection in 150(25.5%) out of 595 skin ulcer samples obtained from 426 study participants. Given that 445(74,8%) of the skin ulcers tested negative for Leishmania parasite suggests a need for investigation of additional causes of skin ulcers such as yaws and Buruli ulcer in the study area.

The main malaria vectors in sub-Saharan Africa, the Anopheles gambiae (Giles; Diptera: Culicidae), normally breed in clean water sources. However, evidence suggests an on-going adaptation of Anopheline species to polluted breeding habitats in urban settings. This study aimed at understanding the adaptation to breeding in water bodies with different qualities, in five selected mosquito breeding sites in urban Accra, Ghana. The study sites were also evaluated for the WHO water-quality parameters as a measure of pollution, and insecticide residues. Field mosquitoes were evaluated for five genes; CYP6P3, CYP4H19, CYP4H24, GSTD1-4, and ABCC11-associated with insecticide detoxification-using quantitative RT-PCR, as well as Mono-oxygenase, Alpha Esterase, Glutathione S-transferase, and insensitive acetylcholinesterase (AChE) using biochemical enzyme assays. The lab-reared, insecticide susceptible An. gambiae Kisumu strain was bred in the most polluted water source for 10 generations and evaluated for the same genes and enzymes. The results revealed that the fold expression of the genes was higher in the larvae compared with the adults. The results also suggest that detoxification enzymes could be involved in the adaptation of An. gambiae to polluted breeding sites. Correlation analysis revealed a highly positive significant correlation between calcium levels and all five genes (P < 0.05). Stepwise linear regression to understand which of the variables predicted the expression of the genes revealed that sulphate was responsible for ABCC11 and CYP4H24, alkalinity for GSTD1-4, and calcium for CYP4H19 and CYP6P3. The detailed genetic basis of this adaptation need to be further investigated. A further understanding of this adaptation may provide outlooks for controlling malaria and other disease vectors adapted to polluted breeding water sources.

Background Malaria vector control methods involving the use of pyrethroids remain the strategies being used against malaria vectors in Ghana. These methods include the use of long-lasting insecticidal nets and indoor residual spraying in many areas in Ghana. However, there is evidence that pyrethroid resistance is widespread in many areas in Ghana. Synergists have been shown to be useful in inhibiting the enzymes that are responsible for the development of resistance and hence enhance the insecticide susceptibility of Anopheles gambiae sensu lato (s.l.) in many areas. The present study investigated the effect of piperonyl butoxide (PBO) on the susceptibility status of An. gambiae s.l. across some sentinel sites in Ghana. Methods Three to five day old An. gambiae s.l. reared from larvae were used in WHO susceptibility tube assays. Batches of 20–25 female adult An. gambiae s.l. were exposed simultaneously to the insecticide alone and to the PBO + insecticide. The knock down rate after 60 min and mortality at 24 h were recorded. Results Deltamethrin and permethrin resistance of An. gambiae s.l. was observed in all the sites in 2015 and 2016. The mortality after 24 h post exposure for deltamethrin ranged from 16.3% in Weija to 82.3% in Kade, whereas that for permethrin ranged from 3.8% in Gomoa Obuasi to 91.3% in Prestea. A significant increase in susceptibility to deltamethrin and less to permethrin was observed during both 2015 and 2016 years in most of the sites when An. gambiae s.l. mosquitoes were pre-exposed to PBO. Conclusion Findings from this study showed that the use of PBO significantly enhanced the susceptibility of An. gambiae s.l. mosquitoes in most of the sentinel sites. It is recommended that vector control strategies incorporating PBO as a synergist can be effective in killing mosquitoes in the presence of deltamethrin and permethrin resistance.

Visceral leishmaniasis (VL) is a major problem worldwide and causes significant morbidity and mortality. Existing drugs against VL have limitations, including their invasive means of administration long duration of treatment regimens. There are also concerns regarding increasing treatment relapses as well as the identification of resistant clinical strains with the use of miltefosine, the sole oral drug for VL. There is, therefore, an urgent need for new alternative oral drugs for VL. In the present study, we show the leishmanicidal effect of a novel, oral antimalarial endoperoxide N-251. In our In vitro studies, N-251 selectively and specifically killed Leishmania donovani D10 amastigotes with no accompanying toxicity toward the host cells. In addition, N-251 exhibited comparable activities against promastigotes of L. donovani D10, as well as other L. donovani complex parasites, suggesting a wide spectrum of activity. Furthermore, even after a progressive infection was established in mice, N-251 significantly eliminated amastigotes when administered orally. Finally, N-251 suppressed granuloma formation in mice liver through parasite death. These findings indicate the therapeutic effect of N-251 as an oral drug, hence suggest N-251 to be a promising lead compound for the development of a new oral chemotherapy against VL., Author summary Visceral Leishmaniasis remains a serious health problem in many developing countries with thousands of new cases recorded annually. Novel oral therapies are required as existing drugs are limited by their invasive means of administration long duration of treatment regimens. Moreover, with miltefosine as the sole oral drug, there are concerns of the eventual development of parasite resistance with its continuous use. In this study, we report on the in vitro and in vivo leishmanicidal effect of orally administered N-251 on the Leishmania donovani complex parasites in mice. Our results suggest that N-251 may be a potential lead compound for the development of a new oral chemotherapy against VL

Background Mass drug administration (MDA) programmes for the control of lymphatic filariasis in Ghana, have been ongoing in some endemic districts for 16 years. The current study aimed to assess factors that govern the success of MDA programmes for breaking transmission of lymphatic filariasis in Ghana. Methods The study was undertaken in two “hotspot” districts (Ahanta West and Kassena Nankana West) and two control districts (Mpohor and Bongo) in Ghana. Mosquitoes were collected and identified using morphological and molecular tools. A proportion of the cibarial armatures of each species was examined. Dissections were performed on Anopheles gambiae for filarial worm detection. A questionnaire was administered to obtain information on MDA compliance and vector control activities. Data were compared between districts to determine factors that might explain persistent transmission of lymphatic filariasis. Results High numbers of mosquitoes were sampled in Ahanta West district compared to Mpohor district (F = 16.09, P = 0.002). There was no significant difference between the numbers of mosquitoes collected in Kassena Nankana West and Bongo districts (F = 2.16, P = 0.185). Mansonia species were predominant in Ahanta West district. An. coluzzii mosquitoes were prevalent in all districts. An. melas with infected and infective filarial worms was found only in Ahanta West district. No differences were found in cibarial teeth numbers and shape for mosquito species in the surveyed districts. Reported MDA coverage was high in all districts. The average use of bednet and indoor residual spraying was 82.4 and 66.2%, respectively. There was high compliance in the five preceding MDA rounds in Ahanta West and Kassena Nankana West districts, both considered hotspots of lymphatic filariasis transmission. Conclusions The study on persistent transmission of lymphatic filariasis in the two areas in Ghana present information that shows the importance of local understanding of factors affecting control and elimination of lymphatic filariasis. Unlike Kassena Nankana West district where transmission dynamics could be explained by initial infection prevalence and low vector densities, ongoing lymphatic filariasis transmission in Ahanta West district might be explained by high biting rates of An. gambiae and initial infection prevalence, coupled with high densities of An. melas and Mansonia vector species that have low or no teeth and exhibiting limitation. Electronic supplementary material The online version of this article (10.1186/s40249-019-0520-1) contains supplementary material, which is available to authorized users.

Mass drug administration (MDA) is the current mainstay to interrupt the transmission of lymphatic filariasis. To monitor whether MDA is effective and transmission of lymphatic filariasis indeed has been interrupted, rigorous surveillance is required. Assessment of transmission by programme managers is usually done via serology. New research suggests that xenomonitoring holds promise for determining the success of lymphatic filariasis interventions. The objective of this study was to assess Wuchereria bancrofti infection in mosquitoes as a post-MDA surveillance tool using xenomonitoring. The study was carried out in four districts of Ghana; Ahanta West, Mpohor, Kassena Nankana West and Bongo. A suite of mosquito sampling methods was employed, including human landing collections, pyrethrum spray catches and window exit traps. Infection of W. bancrofti in mosquitoes was determined using dissection, conventional and real-time polymerase chain reaction and loop mediated isothermal amplification assays. Aedes, Anopheles coustani, An. gambiae, An. pharoensis, Culex and Mansonia mosquitoes were sampled in each of the four study districts. The dissected mosquitoes were positive for filarial infection using molecular assays. Dissected An. melas mosquitoes from Ahanta West district were the only species found positive for filarial parasites. We conclude that whilst samples extracted with Trizol reagent did not show any positives, molecular methods should still be considered for monitoring and surveillance of lymphatic filariasis transmission.

Background Ghana started its national programme to eliminate lymphatic filariasis (LF) in 2000, with mass drug administration (MDA) with ivermectin and albendazole as main strategy. We review the progress towards elimination that was made by 2016 for all endemic districts of Ghana and analyze microfilaria (mf) prevalence from sentinel and spot-check sites in endemic districts. Methods We reviewed district level data on the history of MDA and outcomes of transmission assessment surveys (TAS). We further collated and analyzed mf prevalence data from sentinel and spot-check sites. Results MDA was initiated in 2001–2006 in all 98 endemic districts; by the end of 2016, 81 had stopped MDA after passing TAS and after an average of 11 rounds of treatment (range 8–14 rounds). The median reported coverage for the communities was 77–80%. Mf prevalence survey data were available for 430 communities from 78/98 endemic districts. Baseline mf prevalence data were available for 53 communities, with an average mf prevalence of 8.7% (0–45.7%). Repeated measurements were available for 78 communities, showing a steep decrease in mean mf prevalence in the first few years of MDA, followed by a gradual further decline. In the 2013 and 2014 surveys, 7 and 10 communities respectively were identified with mf prevalence still above 1% (maximum 5.6%). Fifteen of the communities above threshold are all within districts where MDA was still ongoing by 2016. Conclusions The MDA programme of the Ghana Health Services has reduced mf prevalence in sentinel sites below the 1% threshold in 81/98 endemic districts in Ghana, yet 15 communities within 13 districts (MDA ongoing by 2016) had higher prevalence than this threshold during the surveys in 2013 and 2014. These districts may need to intensify interventions to achieve the WHO 2020 target., Author summary Lymphatic filariasis (LF) control in Ghana has relied on ivermectin and albendazole since the year 2000 when the Ghana Filariasis Elimination Programme started. We analyzed trends in microfilaraemia prevalence during MDA, reported coverage, and transmission assessment survey using data obtained from the Ghana Health Services (GHS). The median reported treatment coverage varied between 77–80% over the years. Our results show that the treatment in Ghana made a significant impact in reducing infections

Background Massangam health district (HD), in the West Region of Cameroon, has received ivermectin mass drug administration (MDA) for 20 years, however there is evidence of continued high transmission of Onchocerca volvulus. In order to better understand the transmission dynamics in the HD and inform intervention strategies there is a need to delineate the boundaries of the suspected area of high transmission within the wider transmission zone. Methodology/Principal findings Parasitological and entomological surveys were conducted to map out the breeding sites of Simulium damnosum and evaluate the prevalence of onchocerciasis in neighbouring communities, including Makouopsap sentinel community. Potential rapids were prospected for identification of S. damnosum larvae and black flies collected to determine infectivity rates. Adults were assessed for the presence of O. volvulus microfilariae through a skin snip biopsy and examined for the presence of nodules. Anti Ov-16 antibodies were tested for in children. Four perennial breeding sites were identified on the Rivers Mbam and Nja. Large number of flies were collected along the River Mbam, especially in the rainy season, with up to 955 flies per day, suggesting this river is a perennial source of black flies. A total of 0.8% of parous flies were infective across the study area. Parasitological studies provided evidence of high rates of infection in the sentinel community and three neighbouring communities, with 37.1% of adults microfilariae positive in Makouopsap. High Ov-16 seropositivity in children also provided evidence of recent on-going transmission. In comparison, communities sampled further away from the sentinel community and neighbouring breeding sites were much closer to reaching onchocerciasis elimination targets. Conclusions/Significance This study provides evidence of a particular geographic area of high transmission in an approximate 12 km range around the sentinel community of Makouopsap and the neighbouring breeding sites on the River Nja. To eliminate onchocerciasis by 2025, there is a need to explore alternative intervention strategies in this area of high transmission., Author summary The global goal is to eliminate onchocerciasis by 2025. However, not all areas are on track to do so, as is the case for Massangam health district (HD) in the West Region of Cameroon. A new strategy is required in order to accelerate efforts to reach elimination targets. This study aimed to better understand the transmission dynamics of Onchocerca volvulus in Massangam HD through the identification of black fly (Simulium damnosum) breeding sites, parasitological and entomological studies. The study was able to define an approximate 12 km area of high transmission encompassing four communities with very high levels of infection and evidence of on-going transmission in humans. This area is facilitated by perennial productive black fly breeding sites in the neighbouring rivers of the Nja and Mbam. Multi-faceted interventions are required to target the reservoir of infection facilitating transmission. We suggest the need for a new test and treat strategy in the area of high transmission (with a drug that kills the adult O. volvulus worm living in humans) and larviciding of the breeding sites of the black fly. In areas surrounding the area, increase distribution of ivermectin from annual to twice yearly distribution, whilst also using innovative strategies to improve compliance.

BackgroundWolbachia are important endosymbionts of filarial parasites. The Wolbachia of Onchocerca volvulus, the filarial pathogen responsible for the human disease onchocerciasis, is implicated in the immunopathology of the disease and may be associated with disease severity dependent on the density of Wolbachia. However, little is known in regards to the density and heterogeneity of Wolbachia in microfilariae, the life stage that is thought to be responsible for the pathology.ResultsWe used a real-time qPCR relative copy number assay to estimate the number of Wolbachia genome(s) per nuclear genome of skin microfilariae (Mf), vector L1 and iL3, and nodulectomy adult male and female O. volvulus worms sampled in Ghana and the Democratic Republic of the Congo. Relatively low median Wolbachia copy numbers and variation was observed in the Mf and vector stages, in contrast to significantly higher median and more variable Wolbachia copy number from the iL3 stage to the adult worm stages.ConclusionsThis study provides the first insight into variation in Wolbachia density between the major life stages of the parasite. The relatively invariant ratios observed for Mf and vector stages is in strong contrast to the high degree of variability of Wolbachia to nuclear ratios in adults and may indicate that the mutualistic relationship between the nematode and Wolbachia in these earlier stages is regulated differently, and certainly more stringently, than the relationship in adults.

In this review, we address the interaction between abscisic acid (ABA) and gibberellins (GAs) in regulating non-climacteric fruit development and maturation at the molecular level. We review the interplay of both plant growth regulators in regulating these processes in several fruit of economic importance such as grape berries, strawberry, and citrus, and show how understanding this interaction has resulted in useful agronomic management techniques. We then relate the interplay of both hormones with ethylene and other endogenous factors, such as sugar signaling. We finally review the growing knowledge related to abscisic acid, gibberellins, and the genus Citrus. We illustrate why this woody genus can be considered as an emerging model plant for understanding hormonal circuits in regulating different processes, as most of the finest work on this matter in recent years has been performed by using different Citrus species.