Skin Snips Research Articles

Onchocerca volvulus infection among ivermectin-treated population in a rural setting in Edo State, Nigeria

Background: West Africa includes the most endemic areas in the world with Nigeria being one of the largest countries of West Africa that has been reported to have a high incidence of onchocerciasis. Aim: The aim of this study was to determine Onchocerca volvulus infection among ivermectin-treated population in Ekpan community, Edo State, Nigeria. Setting and Design: A total of 105 participants including 60 males and 45 females with an age range of 15–81 years were recruited in the study. Random sampling technique was employed in the recruitment of participants. Materials and Methods: Two skin snip specimens were taken from the left and right iliac crests of each participant into a microtube containing physiological saline. The skin snips were processed using standard technique and examined microscopically for microfilariae of O. volvulus. Statistical Analysis: The data collected were analyzed using the Chi-square (X2) test and odd ratio. The statistical package used was GraphPad INSTAT. Results: An overall prevalence of 74.3% of O. volvulus infection was observed among ivermectin-treated subjects in Ekpan. Age, occupation, noncompliance of ivermectin treatment, and source of water significantly affected the prevalence of O. volvulus infection among ivermectin-treated population in Ekpan (P < 0.05). Conclusion: Effective distribution of ivermectin and strict compliance of ivermectin treatment are advocated as these efforts will go a long way at controlling as well as reducing the effects of onchocerciasis in this community.

Effect of Ivermectin Treatment on the Frequency of Seizures in Persons with Epilepsy Infected with Onchocerca volvulus.

A clinical trial performed in the Democratic Republic of Congo (DRC), among persons with epilepsy (PWE) infected with Onchocerca volvulus treated with anti-seizure medication suggested that ivermectin reduces the seizure frequency. We assessed the effect of ivermectin treatment on seizure frequency in PWE with and without anti-seizure medication in three onchocerciasis endemic areas (Maridi, South Sudan; Aketi, DRC; and Mahenge, Tanzania). Pre- and 3–5 months post-ivermectin microfilariae densities in skin snips and seizure frequency were assessed. After ivermectin, the median (IQR) percentage reduction in seizure frequency in the study sites ranged from 73.4% (26.0–90.0) to 100% (50.0–100.0). A negative binomial mixed model showed that ivermectin significantly reduced the seizure frequency, with a larger decrease in PWE with a high baseline seizure frequency. Mediation analysis showed that ivermectin reduced the seizure frequencies indirectly through reduction in microfilariae densities but also that ivermectin may have a direct anti-seizure effect. However, given the short half-life of ivermectin and the fact that ivermectin does not penetrate the healthy brain, such a direct anti-seizure effect is unlikely. A randomized controlled trial assessing the ivermectin effect in people infected with O. volvulus who are also PWE on a stable anti-seizure regimen may be needed to clarify the causal relationship between ivermectin and seizure frequency.

Morphological characteristics of microfilariae in blood smears of the common treeshrew Tupaia glis (Mammalia: Scandentia) in Gemas, Negeri Sembilan, Malaysia.

Some filarial nematodes, such as Wuchereria bancrofti, Brugia malayi, and Brugia timori, cause lymphatic diseases in humans in the tropics, whereas other filarial parasites from wild animals cause zoonotic diseases in humans worldwide. To elucidate the prevalence and diversity of filarial parasites in Malaysia, we investigated the filarial parasites from wild animals in Gemas, Negeri Sembilan. To find adult filarial parasites, we dissected 26 animals, which included five frogs, one skink, one snake, two birds, six common treeshrews, and 11 rats. Then, we examined microfilariae in the blood smears and skin snips obtained from each animal. We found two types of microfilariae in the blood smears of common treeshrews: one was very similar to Malayfilaria sofiani and the other closely resembled Brugia tupaiae. These findings indicate an additional distribution of these filarial parasites in Gemas.

Comparison of some immunological parameters between untreated and ivermectin-treated onchocerciasis patients in the Nkwanta North District of Ghana

Background: Onchocerciasis (river blindness) is a neglected tropical disease that affects the skin and the eye. It is caused by a nematode parasite, Onchocerca volvulus. The disease pathology is largely attributed to host immune responses to the microfilariae. The efficacy of ivermectin (IVM) is defined by its microfilaricidal effects (clearing >60% of microfilariae within 24 h, ∼100% within 7 days) and its ability to temporarily retard the embryogenesis of the adult female parasite at least within 90 days. We sought to profile and compare immune parameters of untreated microfilaremic and IVM-treated amicrofilaremic onchocerciasis patients to identify immune factors differentially associated with microfilaremia. This will help understand the disease mechanism. Methods & Materials: Four bloodless skin biopsies were taken from the two iliac crests and two calves using the corneoscleral punch. The microfilariae emerging from the skin biopsies following overnight incubation in physiological saline in 96-well plate was quantified under the inverted light microscope. We measured white blood cell differentials for 124 adults; 53 microfilaremic (MF) and 71 ivermectin treated (1–5 rounds) seropositive amicrofilaremic (AMF) individuals 3 months following last treatment at the Nkwanta North District of Ghana. Subgroups (16 ≤ n ≤ 20) of both MF and first-time IVM-treated AMF were assayed for plasma cytokines, total serum IgE and urine histamine. Comparisons were made by Mann-Whitney U tests. Results: The geometric mean microfilariae density/milligram skin snip was 22.5 (range 5.8–56.3). Basophil count was lower (p < 0.0001) while eosinophil counts were higher (p = 0.02) for MF compared to AMF. Out of 14 cytokines (IL-1β, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12, IL-13, IFN-γ, TNF-α, GM-CSF and TGFβ); three, Interleukin (IL)-13 (p = 0.002), IL-8 (p = 0.03) and Interferon-gamma (p = 0.003) were higher among MF compared to first time IVM-treated AMF. Histamine and IgE were not different between the two groups. Conclusion: IL-13, IL-8 and interferon-gamma, basophils and eosinophils differentiated between untreated and treated patients and hence these immune parameters might be contributing to the disease pathophysiology among microfilaremic onchocerciasis patients in the Nkwanta North District of Ghana.

Point prevalence mapping reveals hotspot for onchocerciasis transmission in the Ndikinimeki Health District, Centre Region, Cameroon

BackgroundIvermectin-based preventive chemotherapy (PC) is distributed annually to all at-risk populations eligible for ivermectin treatment to control and/or eliminate onchocerciasis. Information on the impact of mass ivermectin administration on onchocerciasis transmission is scanty, and it is tricky to appreciate the progress towards elimination and engage corrective measures. To fill that gap in the Centre Region in Cameroon, the current onchocerciasis endemicity level in the Ndikinimeki Health District after about two decades of mass treatments was assessed.MethodsA cluster-based cross-sectional survey was carried out in the Ndikinimeki Health District and all volunteers aged ≥ 5 years were (i) interviewed on their compliance to ivermectin over the past five years and (ii) underwent clinical (nodule palpation and visual search for onchocercal lesions) and parasitological examinations (skin snip) for onchocerciasis.ResultsThe overall Onchocerca volvulus prevalence was 7.0% (95% CI: 5.2–9.3%). The prevalence of the disease was significantly higher in the communities Kiboum 1 and Kiboum 2 compared to the other communities (highest prevalence in Makénéné Town Water: 8.5%; 95% CI: 2.3–20.4%) (χ2 = 51.314, df = 11, P = 0.0001). The proportion of systematic non-compliers to ivermectin was 23.3% (95% CI: 19.9–27.1%) among individuals interviewed. In the sentinel sites (Kiboum communities), onchocerciasis prevalence decreased from 95.2% (95% CI: 88.3–98.1%) to 23.7% (95% CI: 14.7–36.0%).ConclusionsThis study has revealed that the Ndikinimeki Health District is hypo-endemic for onchocerciasis after about two decades of preventive chemotherapy. However, transmission is ongoing, with potential hotspots in the Kiboum 1 and Kiboum 2 communities, which are known as first-line communities (closest to the breeding sites of the vector). Alternative or complementary strategies to annual ivermectin appear compulsory to accelerate the momentum towards onchocerciasis elimination.

Differential susceptibility of Onchocerca volvulus microfilaria to ivermectin in two areas of contrasting history of mass drug administration in Cameroon: relevance of microscopy and molecular techniques for the monitoring of skin microfilarial repopulation within six months of direct observed treatment

BackgroundIvermectin is an excellent microfilaricide against Onchocerca volvulus. However, in some regions, long term use of ivermectin has resulted in sub-optimal responses to the treatment. More data to properly document the phenomenon in various contexts of ivermectin mass drug administration (IVM-MDA) is needed. Also, there is a need to accurately monitor a possible repopulation of skin by microfilariae following treatment. Skin snip microscopy is known to have a low sensitivity in individuals with light infections, which can be the case following treatment. This study was designed with two complementary objectives: (i) to assess the susceptibility of O. volvulus microfilariae to ivermectin in two areas undergoing IVM-MDA for different lengths of time, and (ii) to document the repopulation of skin by the O. volvulus microfilariae following treatment, using 3 independent diagnostic techniques.MethodIdentified microfilaridermic individuals were treated with ivermectin and re-examined after 1, 3, and 6 months using microscopy, actin real-time PCR (actin-qPCR) and O-150 LAMP assays. Susceptibility to ivermectin and trends in detecting reappearance of skin microfilariae were determined using three techniques. Microscopy was used as an imperfect gold standard to determine the performance of actin-qPCR and LAMP.ResultsIn Bafia with over 20 years of IVM-MDA, 11/51 (21.6%) direct observe treated microfilaridemic participants were still positive for skin microfilariae after 1 month. In Melong, with 10 years of IVM-MDA, 2/29 (6.9%) treated participants were still positive. The microfilarial density reduction per skin biopsy within one month following treatment was significantly lower in participants from Bafia.In both study sites, the molecular techniques detected higher proportions of infected individuals than microscopy at all monitoring time points. LAMP demonstrated the highest levels of sensitivity and real-time PCR was found to have the highest specificity.ConclusionPatterns in skin mirofilariae clearance and repopulation were established. O. volvulus worms from Bafia with higher number of annual MDA displayed a lower clearance and higher repopulation rate after treatment with ivermectin. Molecular assays displayed higher sensitivity in monitoring O. volvulus microfilaridemia within six months following treatment.

Clinical Signs of River Blindness and the Efficacy of Ivermectin Therapy in Idogun, Ondo State, Nigeria

River blindness is a skin, and an eye disease caused by Onchocerca volvulus and vectored by a female hematophagous blackfly. The study aims to evaluate the distribution of the clinical signs of river blindness and the efficacy of Ivermectin in the treatment of river blindness in Idogun. Observational studies in epidemiology which involve the use of structured questionnaire to obtain useful epidemiological information from the respondents, physical assessment via palpation from head to ankle was used to assess clinical signs from the respondents and skin snip test was used to evaluate the prevalence of the disease.

Status of parasitological indicators and morbidity burden of onchocerciasis after years of successive implementation of mass distribution of ivermectin in selected communities of Yeki and Asosa districts, Ethiopia

BackgroundControl and elimination of onchocerciasis requires regular follow-up and evaluation of community directed treatment with ivermectin (CDTi) program implementation. This research was aimed to assess the epidemiological status of onchocerciasis in disease endemic communities of Asosa and Yeki districts of Ethiopia after 5 and 15 years of successive CDTi respectively, and to evaluate the decline in infection and morbidity burden.MethodsA community-based cross-sectional study was conducted from August 2017 to January 2018 (i.e. within 2–7 months since the last treatment) using interview, physical and parasitological examinations. Pre-CDTi epidemiological data were obtained from studies conducted prior to the launch of CDTi.ResultsA total of 3002 individuals (1567 from Asosa and 1435 from Yeki) were included. No infection was detected from Yeki. In Asosa, the prevalence of infection was 1.6%. The geometric mean intensity of infection was 0.02 mf/mg of skin snip. The prevalence rates of dermatitis, depigmentation, nodule, and atrophy in Yeki were 33(2.3%), 57(4%), 37(2.6%) and 11(0.7%), respectively. The prevalence rates of papular dermatitis, depigmentation, palpable nodule, atrophy, and blindness in Asosa were 94(6%), 38(2.4%), 30(1.9%), 28(1.8%) and 2(0.1%), respectively. Five years of CDTi had significantly reduced prevalence and intensity of infection by 91.8% (p < 0.001) and 99.7% (p < 0.001), respectively. Moreover, CDTi reduced prevalence of papular dermatitis by 95.9% (p < 0.001), palpable nodule by 90.5% (p < 0.001), and atrophy by 30% (p = 0.6) in Yeki. Similarly, CDTi reduced prevalence of papular dermatitis by 88.6% (p < 0.001), depigmentation by 90.3% (p < 0.001), atrophy by 89.5% (p < 0.001), and blindness by 90% (p < 0.001) in Asosa.ConclusionsFifteen years of successive CDTi had brought the infection from high to zero in Yeki. However, thorough entomological and serological data need to be generated to ascertain whether complete interruption of parasite transmission has been attained, and for considerations of an evidence-based CDTi cessation. Five years of CDTi in Asosa has significantly reduced the infection and morbidity of onchocerciasis to very low level. We, hereby, recommend biannual CDTi to continue in Asosa and its surroundings until the infection transmission is fully interrupted.

Ivermectin Treatment Response in Onchocerca Volvulus Infected Persons with Epilepsy: A Three-Country Short Cohort Study.

Despite a long history of community-directed treatment with ivermectin (CDTI), a high ongoing Onchocerca volvulus transmission is observed in certain onchocerciasis-endemic regions in Africa with a high prevalence of epilepsy. We investigated factors associated with higher microfilarial (mf) density after ivermectin treatment. Skin snips were obtained from O. volvulus-infected persons with epilepsy before, and 3 to 5 months after ivermectin treatment. Participants were enrolled from 4 study sites: Maridi (South Sudan); Logo and Aketi (Democratic Republic of Congo); and Mahenge (Tanzania). Of the 329 participants, 105 (31.9%) had a post-treatment mf density >20% of the pre-treatment value. The percentage reduction in the geometric mean mf density ranged from 69.0% (5 months after treatment) to 89.4% (3 months after treatment). A higher pre-treatment mf density was associated with increased probability of a positive skin snip after ivermectin treatment (p = 0.016). For participants with persistent microfiladermia during follow-up, a higher number of previous CDTI rounds increased the odds of having a post-treatment mf density >20% of the pre-treatment value (p = 0.006). In conclusion, the high onchocerciasis transmission in the study sites may be due to initially high infection intensity in some individuals. Whether the decreasing effect of ivermectin with increasing years of CDTI results from sub-optimal response mechanisms warrants further research.

Comparison of Diagnostic Tests for Onchocerca volvulus in the Democratic Republic of Congo.

Onchocerciasis is diagnosed by detecting microfilariae in skin snips or by detecting OV16 IgG4 antibodies in blood by either enzyme linked immunosorbent assay (ELISA) or a rapid diagnostic test (RDT). Here, we compare the sensitivity and specificity of these three tests in persons with epilepsy living in an onchocerciasis endemic region in the Democratic Republic of Congo. Skin snips and blood samples were collected from 285 individuals for onchocerciasis diagnosis. Three tests were performed: the OV16 RDT (SD Bioline) and the OV16 ELISA both on serum samples, and microscopic detection of microfilariae in skin snips. The sensitivity and specificity of each test was calculated with the combined other tests as a reference. Microfilariae were present in 105 (36.8%) individuals, with a median of 18.5 (6.5–72.0) microfilariae/skin snip. The OV16 RDT and OV16 ELISA were positive in, respectively, 112 (39.3%) and 143 (50.2%) individuals. The OV16 ELISA had the highest sensitivity among the three tests (83%), followed by the OV16 RDT (74.8%) and the skin snip (71.4%). The OV16 RDT had a higher specificity (98.6%) compared to the OV16 ELISA (84.8%). Our study confirms the need to develop more sensitive tests to ensure the accurate detection of ongoing transmission before stopping elimination efforts.

Persons with onchocerciasis-associated epilepsy and nodding seizures have a more severe form of epilepsy with more cognitive impairment and higher levels of Onchocerca volvulus infection.

Following previous reports of very high epilepsy prevalence in the onchocerciasis-endemic villages in Maridi County, South Sudan, a study was conducted to investigate the association between the level of Onchocerca volvulus infection, epilepsy, and related outcomes. In December 2018, persons with epilepsy (PWE) were recruited from villages where an epilepsy prevalence of 4.4% (range: 3.5-11.9%) was documented. We enrolled 318 participants from whom two skin snips were taken for microscopic detection of O. volvulus microfilariae (mf). Seizure history was obtained for all PWE and their degree of disability assessed using the modified Rankin scale. Almost all (84.9%) PWE had detectable mf in their skin snips. Onchocerciasis-infected PWE experienced nodding seizures more often than uninfected PWE (p=0.034). Moreover, persons with nodding seizures had more frequent seizures (p<0.001) and higher disability scores (p<0.001), and were more often cognitively impaired and younger at the time of their first epileptic seizure (nine years vs 12 years, p<0.001) compared to PWE without nodding seizures. Based on multivariate models, nodding seizures were associated with higher mf densities (aOR: 1.022; 95%CI: 1.005-1.041). Epilepsy onset at a younger age was associated with a worse outcome. Higher frequency of seizures, longer duration of epilepsy and younger age were associated with increased disability. Regular antiepileptic drug use was associated with better cognitive and disability outcomes. PWE with nodding seizures have a more severe form of onchocerciasis-associated epilepsy, with earlier seizure onset and higher levels of O. volvulus infection. Younger PWE were prone to worse epilepsy outcomes, which would be prevented with regular antiepileptic treatment.

A Second Population-Based Cohort Study in Cameroon Confirms the Temporal Relationship Between Onchocerciasis and Epilepsy.

To confirm our earlier evidence of a temporal and dose–response relationship between onchocerciasis and epilepsy, we conducted another cohort study in a different setting in Cameroon. Individuals whose Onchocerca volvulus microfilarial density (Ov-MFD) was measured in 1992–1994 when they were children were revisited in 2019 to determine if they acquired epilepsy. With reference to individuals with no microfilariae in 1992–1994, the relative risks of acquiring epilepsy were 0.96, 2.76, 3.67, and 11.87 in subjects with initial Ov-MFD of 1–7, 8–70, 71–200, and > 200 microfilariae per skin snip, respectively. This study further demonstrates reproducibility using the Bradford Hill’s criteria for causality.

The insufficiency of circulating miRNA and DNA as diagnostic tools or as biomarkers of treatment efficacy for Onchocerca volvulus

Skin snip evaluation for onchocerciasis has insufficient sensitivity when skin microfilarial (mf) densities are low, such as following ivermectin treatment. Mf density is suitable for assessing microfilaricidal efficacy but only serves as an indirect indicator of macrofilaricidal activity. We assessed circulating nucleic acids from Onchocerca volvulus as an alternative to skin snips. We screened a plasma sample set of infected individuals followed up at four, 12 and 21 months after microfilaricidal (ivermectin, n = four), macrofilaricidal (doxycycline, n = nine), or combination treatment (n = five). Two parasite-derived miRNAs, cel-miR-71-5p and bma-lin-4, and O-150 repeat DNA were assessed. Highly abundant DNA repeat families identified in the O. volvulus genome were also evaluated. miRNAs were detected in two of 72 plasma samples (2.8%) and two of 47 samples (4.3%) with microfilaridermia using RT-qPCR. O-150 DNA was detected in eight (44.4%) baseline samples by qPCR and the number of positives declined post-treatment. One doxycycline-treated individual remained O-150 positive. However, only 11 (23.4%) samples with microfilaridermia were qPCR-positive. Analysis by qPCR showed novel DNA repeat families were comparatively less abundant than the O-150 repeat. Circulating parasite-derived nucleic acids are therefore insufficient as diagnostic tools or as biomarkers of treatment efficacy for O. volvulus.

Prevalence of onchocerciasis, attitudes and practices and the treatment coverage after 15 years of mass drug administration with ivermectin in the Tombel Health District, Cameroon.

IntroductionOnchocerciasis is an infection caused by Onchocerca volvulus. It affects 37 million people of which 99% are in Africa. This study assessed the prevalence of onchocerciasis after 15 years of mass drug administration with ivermectin.MethodsThis was a population based cross sectional study. Questionnaires covering participants' identity and attitudes and practices of community respondents towards ivermectin were administered. The treatment coverage was obtained by review of records of mass drug administration from 1999 to 2015. The epidemiological evaluation of infection status was done by parasitological examination of skin snips and nodule palpation in individuals in five health areas of the district.ResultsA total of 400 participants were randomly selected. Of these, 56.0% were males, 62.0% single, 59.5% farmers and 98.0% Christians. Participants with good attitudes towards community directed treatment with ivermectin made up 80.5% while 47.8% of the participants had good practice. The highest treatment coverage achieved was 88.0% in 2010 while lowest was 57.0% in 2002. Less than 2% had microfilaria and 6.0% had nodules. There was no statistically significant difference in the prevalence of microfilaria with respect to age. There was a statistically significant difference in the distribution of nodules (χ2=73.6, p=0.001) among the different age groups. The greatest rate of infection (2.1%) was among farmers.ConclusionThis study showed that the prevalence reduced compared to other prevalence studies in Cameroon. The study area was hypo-endemic for onchocerciasis.

The Galabat-Metema cross-border onchocerciasis focus: The first coordinated interruption of onchocerciasis transmission in Africa.

BackgroundOnchocerciasis transmission across international borders is not uncommon, yet a coordinated cross border stops mass drug administration (MDA) decision has not been documented.Methods/Principle findingsThe Galabat-Metema focus involves neighboring districts on the border between Sudan and Ethiopia. Mass drug administration (MDA) was provided once and subsequently twice per year in this focus, with twice-per-year beginning in Ethiopia’s Metema subfocus in 2016 and in the Sudan’s Galabat subfocus in 2008. Ov16 ELISA-based serosurveys were conducted in 6072 children under 10 years of age in the Metema subfocus in 2014, and 3931 in the Galabat in 2015. Between 2014 and 2016, a total of 27,583 vector Simulium damnosum flies from Metema and 9,148 flies from Galabat were tested by pool screen PCR for Onchocerca volvulus O-150 DNA. Only 8 children were Ov16 seropositive (all in the Metema subfocus); all were negative by skin snip PCR. The upper limit of the 95% confidence interval (UCL) for Ov16 seropositive was <0.1% for the overall focus and 0.14 positive fly heads per 2000 (UCL = 0.39/2000). However, an entomological ‘hotspot’ was detected on the Wudi Gemzu river in Metema district. The hotspot was confirmed when 4 more positive fly pools were found on repeat testing in 2017 (1.04 L3/2000 flies (UCL = 2.26/2000). Information exchange between the two countries led to stopping MDA in a coordinated fashion in 2018, with the exception of the hotspot at Wudi Gemzu, where MDA with ivermectin was increased to every three months to hasten interruption of transmission.ConclusionCoordinated stop MDA decisions were made by Sudan and Ethiopia based on data satisfying the World Health Organization’s criteria for interruption of onchocerciasis transmission. Definitions of entomological ‘hotspots’ and buffer zones around the focus are proposed.

Onchocerca volvulus is not detected in the cerebrospinal fluid of persons with onchocerciasis-associated epilepsy

ObjectivesEpidemiological evidence links onchocerciasis with the development of epilepsy. The aim of this study was to detect Onchocerca volvulus microfilariae or its bacterial endosymbiont, Wolbachia, in the cerebrospinal fluid (CSF) of persons with onchocerciasis-associated epilepsy (OAE). MethodsThirteen persons with OAE and O. volvulus skin snip densities of >80 microfilariae were recruited in Maridi County (South Sudan) and their CSF obtained. Cytospin centrifuged preparations of CSF were examined by light microscopy for the presence of O. volvulus microfilariae. DNA was extracted from CSF to detect O. volvulus (O–150 repeat) by quantitative real-time PCR, and Wolbachia (FtsZ gene) by standard PCR. To further investigate whether CSF from onchocerciasis-infected participants could induce seizures, 3- and 7-day old zebrafish larvae were injected with the CSF intracardially and intraperitoneally, respectively. For other zebrafish larvae, CSF was added directly to the larval medium. ResultsNo microfilariae, parasite DNA, or Wolbachia DNA were detected in any of the CSF samples by light microscopy or PCR. All zebrafish survived the procedures and none developed seizures. ConclusionsThe absence of O. volvulus in the CSF suggests that OAE is likely not caused by direct parasite invasion into the central nervous system, but by another phenomenon triggered by O. volvulus infection.

Onchocerciasis-associated epilepsy in the Democratic Republic of Congo: Clinical description and relationship with microfilarial density.

BackgroundHigh epilepsy prevalence and incidence were observed in onchocerciasis-endemic villages in the Democratic Republic of Congo (DRC). We investigated the clinical characteristics of onchocerciasis-associated epilepsy (OAE), and the relationship between seizure severity and microfilarial density.MethodsIn October 2017, ivermectin-naive persons with epilepsy (PWE) were recruited from onchocerciasis-endemic areas in the Logo health zone in the DRC. Additional PWE were enrolled in the Aketi health zone, where ivermectin had been distributed annually for 14 years. Past medical history, clinical characteristics and skin snips for Onchocerca volvulus detection were obtained from participants. Bivariate and multivariable analyses were used to investigate associations with microfilarial density.ResultsOf the 420 PWE in the Logo health zone, 392 were skin snipped (36.5% positive). Generalized motor seizures were most frequent (392 PWE, 93.3%), and nodding seizures were reported in 32 (7.6%) participants. Twelve PWE (3.1%) presented Nakalanga features. Sixty-three (44.1%) skin snip-positive PWE had a family history of epilepsy, compared to only 82 (32.9%) skin snip-negative PWE (p = 0.027). Eighty-one onchocerciasis-infected PWE were recruited in the Aketi health zone. Positive correlations between seizure frequency and microfilarial density were observed in Logo (Spearman-rho = 0.175; p<0.001) and Aketi (Spearman-rho = 0.249; p = 0.029). In the multivariable model adjusted for age, gender, and previous treatment, high seizure frequency was associated with increasing microfilarial density in Aketi (p = 0.025) but not in Logo (p = 0.148).ConclusionIn onchocerciasis-endemic regions in the DRC, a wide spectrum of seizures was observed. The occurrence of Nodding seizures and Nakalanga features, as well as an association between seizure severity and O. volvulus microfilarial density suggest a high OAE prevalence in the study villages.Trial registrationClinicalTrials.gov NCT03052998.

PO 8576 EFFECT OF IVERMECTIN TREATMENT ON THE FREQUENCY OF SEIZURES IN PERSONS WITH ONCHOCERCIASIS-ASSOCIATED EPILEPSY: PRELIMINARY RESULTS OF A RANDOMISED CLINICAL TRIAL

BackgroundMany studies reported an association between epilepsy and onchocerciasis. Moreover, anecdotal evidence suggests that ivermectin may reduce seizure frequency in persons with onchocerciasis-associated epilepsy (PWOAE). Therefore, we performed a randomised clinical trial among ivermectin-naïve persons with epilepsy in onchocerciasis-endemic villages in the Ituri province, Democratic Republic of the Congo.MethodsPWOAE were randomised in an arm receiving immediate (arm A) or delayed (4 months later) ivermectin treatment (arm B). All participants were receiving anti-epileptic drugs. Inclusion criteria were: age &gt;5 years, signed informed consent, normal neurological development until onset of epilepsy between 5–18 years of age, seizure frequency of ≥2 seizures/month, presence of microfilaria in skin snip and/or antibodies against the Ov16 antigen. Primary study outcome: seizure freedom at month 4; secondary outcome:&gt;50% reduction in seizure frequency at month 4 compared to reported seizure frequency at randomisation. The proposed sample size was 110 PWOAE.Results93 PWOAE, 57 males and 36 females, (mean age 22), were enrolled between October and November 2017. On March 2018, 90 (97%) participant completed their 4th-month evaluation. One serious adverse event was observed during the trial (Steven Johnson reaction caused by phenobarbital). Considering all participants there was no significant difference in outcome between the 2 arms. However, considering participants with presence of microfilariae at enrollment, at month 4, 26/39 (66.6%) in arm A and 20/44 (45.5%) in arm B were seizure free (p=0.05) and a 50% reduction of the number of seizures was observed in 9/39 (23.1% ) in arm A and 7/43 (16.3%) in arm B. (p=0.4).ConclusionIvermectin may have an added value in reducing the frequency of seizures in PWOAE treated with anti-epileptic drugs. However, a larger study is needed to confirm this.

Loa loa Microfilariae in Skin Snips: Consequences for Onchocerciasis Monitoring and Evaluation in L. loa-Endemic Areas.

The specificity of skin snips for onchocerciasis diagnoses is considered to be almost 100%. Our molecular methods revealed that microfilariae emerging from skin snips collected from highly microfilaremic Loa loa-infected individuals were largely misidentified as Onchocerca volvulus. This has important implications for onchocerciasis diagnostic testing in Loa-endemic areas.

Low prevalence of epilepsy and onchocerciasis after more than 20\u2009years of ivermectin treatment in the Imo River Basin in Nigeria

BackgroundHigh epilepsy prevalence and incidence have been reported in areas with high onchocerciasis transmission. Recent findings suggest that proper community-directed treatment with ivermectin (CDTI) is potentially able to prevent onchocerciasis-associated epilepsy (OAE). We assessed the epilepsy prevalence and onchocerciasis transmission in two Nigerian villages following more than 20 years of CDTI.MethodsA cross-sectional door-to-door survey was performed in two villages in the Imo River Basin reported to be mesoendomic for onchocerciasis (Umuoparaodu and Umuezeala). Individuals were screened for epilepsy using a validated 5-item questionnaire. Persons suspected to have epilepsy were examined by a neurologist or a physician with training in epilepsy for confirmation. Onchocerciasis was investigated via skin snip microscopy and rapid diagnostic tests for Ov16 antibodies. Results were compared with previous findings from the Imo river basin.ResultsA total of 843 individuals from 257 households in the two villages were encountered. We detected four persons with epilepsy (PWE) giving a crude epilepsy prevalence of 0.5%. This finding differs from observations reported 14 years ago which showed an epilepsy prevalence of 2.8% in the neighbouring village of Umulolo (P = 0.0001), and 1.2% from 13 villages in the Imo river basin (P = 0.07). The seroprevalence of Ov16 antibodies was found to be 0%. Only 4.6% of skin snips were positive compared to 26.8% in previous surveys (P < 0.0001). Ivermectin mass distribution coverage in the study sites in 2017 was 79.7%.ConclusionsA low epilepsy and onchocerciasis prevalence was observed following more than 20 years of CDTI in the Imo River Basin. Absence of Ov16 antibodies indicates minimal transmission of onchocerciasis. These results contrast with observations from areas of high onchocerciasis transmission, where epilepsy prevalence and incidence remain high. Findings from this study suggest that sustained efforts could eventually achieve elimination of onchocerciasis in these villages.