Prospects for developing efficient targets for the xenomonitoring and control of Simulium damnosum s.l., the major vectors of onchocerciasis in Africa.

Despite the ethical issues concerning the continued use of Human Landing Catches (HLC) to monitor the Simulium damnosum complex for epidemiological monitoring of onchocericasis, few attempts to develop alternatives have been reported. In studies on a wild population of S. damnosum in Burkina Faso, we tested visual targets (different sizes and shapes) and olfactory stimuli (CO2 , and POCA and BG-lure® odour blends) for their ability to attract and collect host seeking blackflies. At each trap, blackflies were caught with appropriately sized electrocuting grids and results from Latin square design tests were compared. Throughout, HLCs captured more blackflies than the targets. Of the traps tested, small targets (0.0625 and 0.5 m2 ) were the most efficient visual lure in terms of the number of S. damnosum captured per unit area 1.7-5× more than larger targets. Overall, results suggested that sticky black targets of horizontal rectangular shape (0.125-0.5 m2 ) and baited with a POCA and/or CO2 mixture could provide a cheap practical field alternative to HLC for onchocerciasis xenomonitoring, subject to confirmation that the design has no inherent bias for certain members of the S. damnosum species complex.

Field sensitivity and specificity of the SD BIOLINE onchocerciasis IgG4 Rapid Diagnostic Test in children <10 years old from endemic areas in Burkina Faso.

Skin biopsies (Skin snips) have historically been the gold standard for the diagnosis of onchocerciasis. However, in low prevalence areas and in areas with successful ivermectin mass drug administration (MDA) programs, skin snips are not sensitive enough to decide when to stop MDA; thus, serological diagnostic tools have been recommended for this purpose. This study assessed the sensitivity and specificity of the Ov16 Rapid Diagnostic Test (SD BIOLINE Onchocerciasis RDT) compared to skin snip in endemic areas undergoing ivermectin mass distribution using Community Directed Treatment with Ivermectin (CDTI) strategy. A cross-sectional study was conducted between September and November 2016 in five endemic villages in the Cascades region in Burkina Faso. Children aged 2 to 9-years were examined during the impact epidemiological survey using both the skin snip and Ov16 Rapid Diagnostic Test. The Ov16 Rapid Diagnostic Test sensitivity and specificity were determined with reference to the skin biopsy. Skin snip positivity was 1.25% in this population, while seroprevalence was 6.5%. When compared to the skin snip as the gold standard, the sensitivity of the Ov16 Rapid Diagnostic Test was 60% and the specificity 94%. When the Ov16 Rapid Diagnostic Test was considered as the gold standard, the skin snip exhibited a sensitivity of 11.5% and a specificity of 99.5%. These results are similar to other studies comparing the performance of the Ov16 ELISA to skin snips, suggesting that the Ov16 RDT may be a useful tool for ivermectin STOP MDA and post transmission surveys, assuming that the prevalence of infection is low or close to zero, and the Ov16 RDT detected also pre patent infections.

Assessment of Culicidae collection methods for xenomonitoring lymphatic filariasis in malaria co-infection context in Burkina Faso.

BACKGROUND: Entomological surveillance of lymphatic filariasis and malaria infections play an important role in the decision-making of national programs to control, or eliminate these both diseases. In areas where both diseases prevalence is low, a large number of mosquitoes need to be sampled to determine vectors infection rate. To do this, efficient mosquito collection methods must be used. This study is part in this framework, to assess appropriate mosquito collection methods for lymphatic filariasis xenomonitoring in a coexistence context with malaria in Burkina Faso. METHODOLOGY/PRINCIPAL FINDINGS: Mosquito collections were performed between August and September 2018 in four villages (Koulpissi, Seiga, and Péribgan, Saptan), distributed in East and South-West health regions of Burkina Faso. Different collection methods were used: Human Landing Catches (HLC) executed indoor and outdoor, Window Exit-Trap, Double Net Trap (DNT) and Pyrethrum Spray Catches (PSC). Molecular analyses were performed to identify Anopheles gambiae s.l. sibling species and to detect Wuchereria bancrofti and Plasmodium falciparum infection in Anopheles mosquitoes. A total of 3 322 mosquitoes were collected among this, Anopheles gambiae s.l. was the vector caught in largest proportion (63.82%). An. gambiae s.l. sibling species molecular characterization showed that An. gambiae was the dominant specie in all villages. The Human Landing Catches (indoor and outdoor) collected the highest proportion of mosquitoes (between 61.5% and 82.79%). For the sampling vectors infected to W. bancrofti or P. falciparum, PSC, HLC and Window Exit-Trap were found the most effective collection methods. CONCLUSIONS/SIGNIFICANCE: This study revealed that HLC indoor and outdoor remained the most effective collection method. Likewise, the results showed the probability to use Window Exit-Trap and PSC collection methods to sample Anopheles infected.

Progress towards elimination of onchocerciasis in the Region du Sud-Ouest of Burkina Faso which was previously subject to a recrudescence event after vector control.

BACKGROUND: The Sud-Ouest region of Burkina Faso (especially the Bougouriba valley) has been historically problematic with respect to onchocerciasis control, with a recrudescence of infections after vector control carried out the WHO Onchocerciasis Control Programme was halted in 1989. After 1996, mass drug administration of ivermectin was instigated to control the recrudescence so that it would no longer constitute a public health problem. However, in 2010 WHO changed its recommended policy from control to elimination, and in 2013 biannual Community-Directed Treatment with Ivermectin (CDTI) was instigated. Epidemiological surveys were carried-out in 2011 and 2018 to determine whether CDTI was producing a decline in infection levels and progress towards elimination. METHODOLOGY/PRINCIPAL FINDINGS: A cross-sectional study was conducted across 20 villages in four health districts in 2011 and 29 villages in 2018. Individuals aged five years and above were examined by skin-snip, and the prevalence and microfilarial load was determined for each village. In 2011, 75% of villages had some infections and 20% had prevalences >5%, with a mean prevalence across all villages of 2.63% (range 0.0-9.7%), and community microfilarial load ranging from 0 to 0.25 microfilariae per biopsy. In 2018, nine villages (= 31% of total) had some infections, with prevalences ranging from 0.41% to 3.54%, and a mean prevalence across all villages of 0.37%. Community microfilarial load ranged from 0 to 0.1. Amongst those people found to be microfilarial positive, 87% had a history of migration. CONCLUSIONS/SIGNIFICANCE: The endemicity of onchocerciasis infection in the Sud-Ouest region has declined to low levels and seems to be progressing towards elimination. Our findings indicated that biannual CDTI is having good effect, but it should continue for a number of years to ensure elimination of transmission. However, progress towards elimination has a troublesome history in this region, and it would be advisable to select more sentinel villages to have confidence in any future epidemiological and entomological surveys, especially Stop-MDA surveys. With positive individuals migrating between countries, cross-border collaboration needs more attention to ensure effective treatment for onchocerciasis elimination.

Vector control and entomological capacity for onchocerciasis elimination.

Mass drug administration (MDA) of ivermectin is currently the main strategy to achieve elimination of transmission (EoT) of onchocerciasis. Modelling suggests that EoT may not be reached in all endemic foci using annual MDA alone. Onchocerciasis and loiasis are coendemic in forest areas of Central Africa where ivermectin treatment can lead to severe adverse events in individuals with heavy loiasis load, rendering MDA inappropriate. Vector control has been proposed as a complementary intervention strategy. Here, we discuss (i) achievements and pitfalls of previous interventions; (ii) epidemiological impact, feasibility, and combination with MDA to accelerate and/or protect EoT; (iii) role of modelling; (iv) opportunities for innovative methods of vector monitoring and control; and (v) strengthening entomological capacity in endemic countries.

Serological and molecular detection of Leishmania species in dog peripheral blood from Bobo-Dioulasso city, a confirmation of canine leishmaniasis enzootic area for Burkina Faso.

Canine leishmaniasis is increasingly reported worldwide and represent a threat to both animal and human health. In a previous pilot study conducted in Bobo-Dioulasso, the second town of Burkina Faso, we reported five cases of canine leishmaniasis. With the perspective of a One Health action plan, and in the context of increasing urbanization, this study aimed to provide new information on Leishmania spp in dogs in this city. A cross-sectional survey was carried out from May to August 2018 in six districts of the city in order to record clinical and biological data from domestic dogs randomly selected per district. Blood samples were collected into EDTA tubes (4-5 mL), treated and stored at -20 °C until further analyses. The infection status of the dogs was performed by serological tests using plasma, and real time-PCR (RT-PCR) to detect Leishmania parasites using buffy coats. Nested PCR was used for typing the Leishmania species in dogs which were found to be RT-PCR positive. A total of 147 dogs were examined clinically and sampled for blood collection, including 53.7% females and 46.3% of males with a median age of 3 years. The seroincidence of Leishmania parasites within this dog population was 4.76% (95% CI:2.26-9.72). The incidence of Leishmania was 10.88% (95% CI: 6.73-17.11) by RT-PCR which was significantly more sensitive (p = 0,047) and a fair concordance was observed between both tests (Kappa = 0.39, p < 0.001). The characterization of Leishmania species revealed that L. major was circulating in this domestic dog population. Our results confirmed the persistence of zoonotic circulation of Leishmania parasites such as L. major currently in Bobo-Dioulasso city and highlight the need for targeted interventions in order to control transmission of leishmaniasis in this region.

The impact of ivermectin on onchocerciasis in villages co-endemic for lymphatic filariasis in an area of onchocerciasis recrudescence in Burkina Faso.

In Burkina Faso, onchocerciasis was no longer a public health problem when the WHO Onchocerciasis Control Programme in West Africa closed at the end in 2002. However, epidemiological surveillance carried out from November 2010 to February of 2011, showed a recrudescence of infection in the Cascades Region. This finding was made at a time when ivermectin, a drug recommended for the treatment of both onchocerciasis and lymphatic filariasis, had been distributed in this area since 2004 for the elimination of lymphatic filariasis. It was surprising that ivermectin distributed for treating lymphatic filariasis had not prevented the recrudescence of onchocerciasis. Faced with this situation, the aim of our study was to evaluate the effectiveness of ivermectin on the onchocerciasis parasite. The percentage reduction in microfilarial load after treatment with ivermectin was used as a proxy measure for assessing possible resistance. A cohort study was carried out with 130 individuals who had tested positive for microfilariae of Onchocerca volvulus in 2010 using microscopic examination of skin-snip biopsies from five endemic villages. Subjects were followed from July 2011 to June 2012. The microfilarial load of each individual was enumerated by skin-snip biopsy in 2010, prior to the first ivermectin treatment against onchocerciasis under community guidelines. All individuals received two ivermectin treatments six months apart. In 2012, the microfilarial loads were determined again, six months after the second round of ivermectin and the reductions in parasite loads were calculated to measure the impact of the drug. The percentage reduction of the microfilarial loads ranged from 87% to 98% in the villages. In all villages, there was a statistically significant difference between the average microfilarial loads in 2010 and 2012. The level of reduction of microfilarial loads suggests that ivermectin is effective against the recrudescent population of O. volvulus in Cascades Region of Burkina Faso. Further investigations would be necessary to determine the causes of the recrudescence of onchocerciasis. (For French language abstract, see S1 Alternative Language Abstract-Translation of the Abstract into French by the authors.).

Monitoring Insecticide Susceptibility in Aedes Aegypti Populations from the Two Biggest Cities, Ouagadougou and Bobo-Dioulasso, in Burkina Faso: Implication of Metabolic Resistance.

In West Africa, Aedes aegypti remains the major vector of dengue virus. Since 2013, dengue fever has been reemerging in Burkina Faso with annual outbreaks, thus becoming a major public health problem. Its control relies on vector control, which is unfortunately facing the problem of insecticide resistance. At the time of this study, although data on phenotypic resistance were available, information related to the metabolic resistance in Aedes populations from Burkina Faso remained very scarce. Here, we assessed the phenotypic and the metabolic resistance of Ae. aegypti populations sampled from the two main urban areas (Ouagadougou and Bobo-Dioulasso) of Burkina Faso. Insecticide susceptibility bioassays to chlorpyriphos-methyl 0.4%, bendiocarb 0.1% and deltamethrin 0.05% were performed on natural populations of Ae. aegypti using the WHO protocol. The activity of enzymes involved in the rapid detoxification of insecticides, especially non-specific esterases, oxidases (cytochrome P450) and glutathione-S-transferases, was measured on individual mosquitos. The mortality rates for deltamethrin 0.05% were low and ranged from 20.72% to 89.62% in the Bobo-Dioulasso and Ouagadougou sites, respectively. When bendiocarb 0.1% was tested, the mortality rates ranged from 7.73% to 71.23%. Interestingly, in the two urban areas, mosquitoes were found to be fully susceptible to chlorpyriphos-methyl 0.4%. Elevated activity of non-specific esterases and glutathione-S-transferases was reported, suggesting multiple resistance mechanisms involved in Ae. aegypti populations from Bobo-Dioulasso and Ouagadougou (including cytochrome P450). This update to the insecticide resistance status within Ae. aegypti populations in the two biggest cities is important to better plan dengue vectors control in the country and provides valuable information for improving vector control strategies in Burkina Faso, West Africa.

Entomological assessment of the transmission following recrudescence of onchocerciasis in the Comoé Valley, Burkina Faso.

BACKGROUND: Onchocerciasis, or river blindness, is a dermal filariasis caused by infection with the nematode parasite Onchocerca volvulus, transmitted to humans through the bites of blackflies of the genus Simulium. Despite the decade-long West African Regional Programme for the Elimination of Onchocerciasis, involving the mass administration of ivermectin to populations in endemic areas, recrudescence has occurred. An example is in the Cascades Region of south-west Burkina Faso where the resumption of transmission had resulted in infection prevalences of up to 70% in some villages. In 2011, a strategy for community-directed distribution of ivermectin (CDTI) was set up to respond to this worrying re-emergence. Here, we report on a study of Onchocerca spp. transmission in the affected area carried out from January to December 2012. Every month, host-seeking adult females of the S. damnosum complex were collected at sites on the River Comoé near the four villages (Bodadiougou, Bolibana, Badara Karaboro and Badara Dogossè) that had recorded the highest prevalences in 2010. Collected blackflies were dissected and infective larvae were identified using the O-150 PCR method. RESULTS: A total of 9114 S. damnosum (s.l.) adult females were collected, of which 5142 were parous (56.4%) and 78 (1.51%) were infective carrying a total of 137 infective larvae. The annual transmission potential (ATP) was calculated as 0, 30, 255 and 771 infective larvae/man/year in Badara Dogossè, Bolibana, Badara Karaboro and Bodadiougou, respectively. Transmission levels in the latter two are of particular concern as they were higher than 100 infective larvae/person/year, the designated minimum threshold required for elimination of severe pathology, including damage to vision. CONCLUSIONS: These results confirm that recrudescence of onchocerciasis has occurred, and that transmission of O. volvulus was active at sites on the Comoé River in the Cascades region in 2012. In accordance with WHO recommendations, CDTI should be continued and the situation in the Cascades region should be closely monitored if further spread of this outbreak is to be avoided.

Onchocerciasis prevalence, human migration and risks for onchocerciasis elimination in the Upper Mouhoun, Nakambé and Nazinon river basins in Burkina Faso.

Historically, the whole of Burkina Faso was considered to be endemic for onchocerciasis (except a small area in the far north of the country) with prevalence rates 60-80%, but all endemic areas were included in the World Health Organisation Onchocerciasis Control Programme, which operated a system of vector control by larviciding beginning in 1974. In Burkina Faso larviciding had been phased out by 1989 when it was considered that onchocerciasis had been reduced to levels below the transmission breakpoint (and any residual infections would disappear without further intervention). There was never any mass drug administration against onchocerciasis in Burkina Faso, except in the Bougouriba and Comoé river basins (from 1996 and 2011 to present respectively) because in each of these two areas there was a resurgence of infection, and in parts of the Nakambé River basin and Sissili River basin from 1992 to 1998. However, mass drug administration with ivermectin was also phased in across the whole country starting in 2000 using ivermectin against lymphatic filariasis and is currently being phased out (depending upon the epidemiological parameters). In this publication we report a new epidemiological survey for onchocerciasis which was carried out in 2014 in the Upper Mouhoun, Nakambé and Nazinon river basins in Burkina Faso to evaluate the prevalence and intensity of infection of onchocerciasis. A total of 11,195 people from 61 villages were examined across these three river basins, and onchocerciasis prevalence by skin-snip was below 5% in all villages, below 1% in 57 villages (93% of 61 villages) and zero in 47. In the 14 villages with positive skin snips, prevalence figures ranged from 0.31% to 3.50%. During the survey 31 infected individuals were found. All of them were Burkinabé, of whom 30 had a recent history of residence in Côte d'Ivoire (with a range of 0.5 to 73 microfilariae per skin-snip from two snips per person) and only one had no history of migration and presumably had an autochthonous infection (mean of 0.5 microfilariae per skin snip from two snips). According to parasitological indicators listed by the World Health Organization African Programme for Onchocerciasis Control in 2010, the situation for onchocerciasis was considered to be satisfactory in all three river basins and probably below the transmission threshold, in which case the disease should disappear naturally without the need for further intervention in the absence of continuing immigration. However, the results clearly indicate that infected persons coming from endemic zones of Côte d'Ivoire are settling in small communities which are otherwise nearly free from onchocerciasis in Burkina Faso. They are thus a source of continuing re-introduction of the parasite into the basins and could be a risk for the achievement of onchocerciasis elimination in all three basins. This would justify the continuation of periodic epidemiological surveys to monitor the possible recrudescence of the disease, and entomological (vector) surveys should be undertaken to assess and monitor the residual transmission.

Recrudescence of onchocerciasis in the Comoé valley in Southwest Burkina Faso.

Onchocerciasis control by vector control was instigated in southwest Burkina Faso in January 1969 by ORSTOM/OCCGE, and continued until operations were taken over by the WHO Onchocerciasis Control Programme (OCP) in February 1975, which itself ceased operations in the area in 1989 when onchocerciasis was judged to have been reduced to insignificant levels. Initially (1969-1975) vector immigration maintained unacceptably high levels of transmission, but OCP was much larger than the preceding campaign and in 1975 the Annual Transmission Potential (ATP) dropped below 100 at all sites in the Comoé river valley except Folonzo, which continued to be subject to reinvasion, along with the whole of the Léraba river valley. However, after the southern extension of the OCP in 1979, ATPs dropped below 100 everywhere in the Comoé basin (including the Léraba valley), and further dropped to insignificant levels after the western extension of the OCP in 1985. Thus transmission dropped more quickly in the Comoé river valley than the Léraba river valley (which had been subject to vector reinvasion), and this was also reflected in prevalence of microfilaraemia in the human population. After 1986 prevalence was less than 5% in all villages in the Comoé river valley (except for two, which subsequently dropped to 0% and 3.7% by 1999). However, in 2001 (12 years after the cessation of vector control) the prevalence in one village in the Comoé river valley had increased to 39.6%, and two more had increased above 5% by 2007. New epidemiological surveys in 2011 and 2012 showed that in 13 out of 30 villages in the Comoé river valley prevalence of microfilaraemia was above 5%, although this was not observed in the Léraba river valley where prevalence remained low. This is the first documented case of recrudescence of onchocerciasis in the old OCP area, and the reasons are not clear. It is possible that there has been immigration of parasites with humans or vectors from areas where there has been a shorter period of control, or that control has been less effective. It is possible that in spite of very low levels of transmission the local parasite population was never reduced to a level below the transmission breakpoint, or that there has been a local recrudescence due to stochastic population effects. In any case it is clear that the distribution of ivermectin against lymphatic filariasis in the area since 2004 has failed to prevent the recrudescence of onchocerciasis, and the Burkina Faso Programme National de Lutte contre l'Onchocercose (PNLO - Ministere de la Santé) has instigated a programme of Community Directed Treatment with Ivermectin specifically aimed at onchocerciasis in accordance with the strategy developed by APOC and recommended to governments by OCP when it was dissolved in 2002.

Optimization of the Esperanza window trap for the collection of the African onchocerciasis vector Simulium damnosum sensu lato.

A simple inexpensive trap (Esperanza window trap) was shown recently to collect significant numbers of Simulium ochraceum sensu lato, a major vector of Onchocerca volvulus in Mesoamerica. Here, we report studies optimizing this trap for the collection of Simulium damnosum s.l., the major vector of O. volvulus in Africa. A shortened, blue and black striped version of the Esperanza window trap, when baited with a combination of CO2 and worn trousers, rivalled human landing collections in the number of S. damnosum s.l. females collected. Traps baited with a commercially available human skin lure and CO2 resulted in collections that were not significantly different than those obtained from traps baited with worn trousers and CO2. This suggests that the Esperanza window trap may offer a replacement for human landing collections for monitoring onchocerciasis transmission in Africa.