Skin Snips Research Articles

O-5S quantitative real-time PCR: a new diagnostic tool for laboratory confirmation of human onchocerciasis

BackgroundOnchocerciasis is a parasitic disease caused by the filarial nematode Onchocerca volvulus. In endemic areas, the diagnosis is commonly confirmed by microscopic examination of skin snip samples, though this technique is considered to have low sensitivity. The available melting-curve based quantitative real-time PCR (qPCR) using degenerated primers targeting the O-150 repeat of O. volvulus was considered insufficient for confirming the individual diagnosis, especially in elimination studies. This study aimed to improve detection of O. volvulus DNA in clinical samples through the development of a highly sensitive qPCR assay.MethodsA novel hydrolysis probe based qPCR assay was designed targeting the specific sequence of the O. volvulus O-5S rRNA gene. A total of 200 clinically suspected onchocerciasis cases were included from Goma district in South-west Ethiopia, from October 2012 through May 2013. Skin snip samples were collected and subjected to microscopy, O-150 qPCR, and the novel O-5S qPCR.ResultsAmong the 200 individuals, 133 patients tested positive (positivity rate of 66.5%) and 67 negative by O-5S qPCR, 74 tested positive by microscopy (37.0%) and 78 tested positive by O-150 qPCR (39.0%). Among the 133 O-5S qPCR positive individuals, microscopy and O-150 qPCR detected 55.6 and 59.4% patients, respectively, implying a higher sensitivity of O-5S qPCR than microscopy and O-150 qPCR. None of the 67 individuals who tested negative by O-5S qPCR tested positive by microscopy or O-150 qPCR, implying 100% specificity of the newly designed O-5S qPCR assay.ConclusionsThe novel O-5S qPCR assay is more sensitive than both microscopic examination and the existing O-150 qPCR for the detection of O. volvulus from skin snip samples. The newly designed assay is an important step towards appropriate individual diagnosis and control of onchocerciasis.

Prevalence and distribution of onchocerciasis in Ahani-Achi Community, Enugu State, Nigeria

The periodic evaluation of onchocerciasis control programmes is paramount in order to ascertain the extent of realization of set goals. Therefore, the demographic pattern of infection in Ahani-Achi Community, Enugu State of Nigeria was investigated from March 2012 to April 2014. Three hundred and twenty-eight (328) participants aged 10->60 years were examined for O. volvulus infection using standard skin snip biopsy technique. Out of this number, 100 (30.49%) tested positive to O. volvulus parasitemia. Females (34.45%) were more infected than males (19.98%), although the difference was not statistically significant (p>0.05). The >60 years age-group was significantly (p<0.05) more infected in females (25.63%) than in males (10%) and farmers were significantly (p<0.05) more infected (40%) than other occupation groups. The study posits that intervention programmes in the area have been reasonably successful but efforts must be sustained towards a possible eradication of the disease.Keywords: Prevalence; distribution; river blindness (onchocerciasis); Ahani-Achi

Sarcoptes scabiei (Acari: Sarcoptidae) infestation in rabbits (Oryctolagus cuniculus): A case study

Sarcoptic mange was suspected in three of five European albino rabbits (Oryctolagus cuniculus) kept for experimental purposes. Gross examination revealed multifocal areas of alopecia around the eyes, nostrils and lips. Skin snips were processed using PCR for the molecular identification of the suspected mites. Histopathology of the skin snips showed erupted epidermis and stratum corneum with an infiltration of inflammatory cells. Skin scraping examination revealed the presence of adult mites as well as eggs. Microscopic taxonomy identified the adult mites as Sarcoptes (S.) scabiei (Acari: Sarcoptidae). The results of PCR indicated a 311 bp band from all the three cases, which confirmed the S. scabiei infestation in rabbits. Sarcoptes scabiei may be a public health concernthrough direct transmission from rabbits infested with S. scabiei through handling. To the best of our knowledge, this is the first report of S. scabiei infection in rabbits from Pakistan.

Evaluation of Onchocerciasis Transmission in Tanzania: Preliminary Rapid Field Results in the Tukuyu Focus, 2015.

To compare diagnostic tests for onchocerciasis in a setting that has suppressed transmission, a randomized, age-stratified study was implemented in an area in Tanzania that had received 15 rounds of annual mass drug administration (MDA) with ivermectin. Study participants (N = 948) from 11 villages underwent a questionnaire, skin examination, skin snips, and blood draw. The burden of symptomatic disease was low. Ov-16 antibody rapid diagnostic test (RDT) results were positive in 38 (5.5%) participants, with 1 (0.5%), 1 (0.4%), and 2 (0.8%) in children aged 0-5, 6-10, and 11-15 years, respectively. Despite significant impact of MDA on transmission, the area would have failed to meet World Health Organization serologic criteria for stopping MDA if a full evaluation had been conducted. The specificity of the RDT, which is 97-98%, may result in the identification of a number of false positives that would exceed the current stop MDA threshold.

Comparing reliability of skin microfilarial load with first internal transcribed spacer primers DNA amplification in monitoring onchocerciasis treatment control

Treatment compliance is a serious challenge to community-directed treatment intervention (CDTI) targeting onchocerciasis eradication. Isolated cases of active infections could jeopardise decision to stop mass drug distribution. Onchocerciasis co-endemicity with lymphatic filariasis in the study-area informed the use of primers capable of differentiating sympatric infections in a single assay. The study-population was hitherto found to be negative for presence of skin microfilaria. DNA extracted from skin snips of patients (n=63) comprised of males, n=21 and females, n=42 were analysed using 18S first internal transcribed spacer (ITS1) in polymerase chain reaction (PCR) assay. Amplified DNA sequences were subjected to basic local alignment search tool. Spectrophotometric analysis of DNA concentrations ranged between 1.5- and 24.5 Nano gram (ng) and the two positive controls (adult worm DNA extract) were 10.5 and 105 ng at 260 nanometer (nm) wave length. The 260/ 280 nm ratios were between 1.44 and 1.68. Four samples (6.35%), two males and two females, and the two positive controls showed PCR amplification products. Two of the positive samples indicated double bands in an agarose gel electrophoregram with molecular weight of 350 and 500 base pairs (bp), while two had single band of 350bp, which is within the expected Mw range of 344 bp for Onchocerca volvulus. The double bands may not be due to unspecific amplification. Moreover, the DNA sequence of a sample had 69.1% homology with control sequence but not with O. volvulus sequences available in the GenBank database. This study underscored the need to assess effectiveness of CDTI with highly sensitive test and not depend on microscopy of emerged microfilaria from skin snips. Whether the DNA was from dead or living microfilaria remained a conjecture. Keywords : Skin microfilaria; DNA sequences.

EFFECT OF ONCHOCERCIASIS TREATMENT ON THE FREQUENCY OF SEIZURES IN PATIENTS WITH EPILEPSY AND ONCHOCERCIASIS

BackgroundA high prevalence of epilepsy is mainly observed in Onchocerca volvulus (OV) hyperendemic areas with no or low ivermectin coverage. There is anecdotal evidence that ivermectin may reduce seizure frequency...

Retinoid Expression in Onchocercal Skin Disease: Pilot Study.

Based on the observation that the parasite Onchocerca volvulus selectively absorbs vitamin A from the host, and the known toxicity of vitamin A in higher concentration, it was hypothesized that dying microfilariae (mf) release their stores of vitamin A (retinoids) into the host circulation in toxic concentrations, inducing the signs and symptoms of onchocerciasis. We conducted a pilot study to test the hypothesis in Songea communities in Southern Tanzania, where mass drug administration with ivermectin had not been implemented by the time of the survey. The specific aim was to evaluate the correlation between the diagnosis of onchocerciasis and increased levels of retinoic acid at infection sites. The analysis was performed by determining copy numbers of a genome of O volvulus present in skin snip samples of persons with onchocerciacis, and correlating these numbers with expression levels of retinoic acid receptor-α (RAR-α), which is inducible by retinoic acid. Total DNA and RNA were extracted from each of 25 mf-positive and 25 mf-negative skin samples and evaluated using quantitative polymerase chain reaction with appropriate negative controls. Analysis of the samples, adjusted with glyceraldehyde 3-phosphate dehydrogenase gene levels, revealed that most samples with detectable RAR-α transcripts had higher levels of RAR-α expression than the assay control. However, the quality and number of samples were insufficient for statistical analysis. Fold data on the expression levels of both O volvulus DNA and RAR RNA suggested a possible trend toward higher relative RAR-α expression in samples with higher levels of O volvulus DNA (r2 = 0.25, P = .079). Evidence of a contribution of vitamin A to the pathology of onchocerciasis thus remains elusive. Future studies on the role of retinoids in onchocerciasis will require larger groups of participants as well as careful monitoring of the cold chain and tissue storage procedures in view of the sensitivity of vitamin A to heat and light.

Nodding syndrome (NS) and Onchocerca Volvulus (OV) in Northern Uganda.

Nodding Syndrome (NS) is a childhood neurological disorder characterized by atonic seizures, cognitive decline, school dropout, muscle weakness, thermal dysfunction, wasting and stunted growth. There are recent published information suggesting associations between Nodding Syndrome (NS) with cerebrospinal fluid (CSF) VGKC antibodies and serum leiomidin-1 antibody cross reacting with Onchocerca Volvulus (OV). These findings suggest a neuro-inflammatory cause of NS and they are important findings in the search for the cause of Nodding Syndrome. These observations perhaps provide further, the unique explanation for the association between Nodding Syndrome and Onchocerca Volvulus. Many clinical and epidemiological studies had shown a significant correlation between NS and infestation with a nematode, Onchocerca volvulus which causes a disease, Onchocerciasis, some of which when left untreated can develop visual defect ("River Blindness"). While these studies conducted in Northern Uganda and Southern Sudan indicate a statistically significant association with (OV infection (using positive skin snips), we observe that (OV is generally endemic in many parts of Sub Saharan Africa and Latin America and that to date, no NS cases have been recorded in those regions. This letter to the Editor is to provide additional information on the current view about the relationship between Nodding Syndrome and Onchocerca Volvulus as seen in Northern Uganda.

Interruption of the transmission of Onchocerca volvulus in the Kashoya-Kitomi focus, western Uganda by long-term ivermectin treatment and elimination of the vector Simulium neavei by larviciding

Uganda is the only country in sub-Saharan Africa whose onchocerciasis elimination programme extensively uses vector control and biannual treatment with ivermectin. The purpose of this study was to assess the impact of combined strategies on interrupting onchocerciasis transmission in the Kashoya-Kitomi focus. Mass Drug Administration annually (13 years) followed by biannual treatments (6 years) and ground larviciding (36 cycles in 3 years) with temephos (Abate®, EC500) against Simulium neavei were conducted. Routine fly catches were conducted for over seven years in six catching sites and freshwater crabs Potamonautes aloysiisabaudiae were examined for immature stages of Simulium neavei. Epidemiological assessments by skin snip were performed in 2004 and 2013. Collection of dry blood spots (DBS) from children <10 years for IgG4 antibodies analysis were done in 2010 and 2013.Treatment coverage with ivermectin improved with introduction of biannual treatment strategy. Microfilaria prevalence reduced from 85% in 1991 to 62% in 2004; and to only 0.5% in 2013. Crab infestation reduced from 59% in 2007 to 0% in 2013 following ground larviciding. Comparison of total fly catches before and after ground larviciding revealed a drop from 5334 flies in 2007 to 0 flies in 2009. Serological assays conducted among 1,362 children in 2010 revealed 11 positive cases (0.8%; 95% CI: 0.4%–1.2%). However, assessment conducted on 3246 children in 2013 revealed five positives, giving point prevalence of 0.15%; 95% CI: 0.02%-0.28%. Four of the five children subjected to O-150 PCR proved negative. The data show that transmission of onchocerciasis has been interrupted based on national and WHO Guidelines of 2012 and 2016, respectively.

Comparing antibody responses to &lt;i&gt;Onchocerca volvulus&lt;/i&gt; and non-parasite antigens in placebo-controlled and ivermectin-treated onchocerciasis patients

Serum antibodies to parasite-specific and non-parasite antigens were evaluated using enzyme-linked immunosorbent assay (ELISA). Out of the 470 sera collected, 409 were from residents of an onchocerciasis hyper-endemic area, 55 non-endemic and 6 European normal sera served as control. The patients’ age, sex, skin microfilaria densities, dermal and ocular clinical manifestations (colour of optic disc) have been well characterised. The study population had participated in a placebocontrolled (n=191) trial of ivermectin (Mectizan®) treatment (n=218). The parasite antigens are phosphate buffered saline crude extract of adult worms of Onchocerca volvulus, a recombinant antigen (Ov1.9) and a monoclonal antibody purified antigen (Cam 1). The non-parasite antigens are deoxycholate citrate extract of optic nerve (nerve-DOC) and commercially available IgA, IgM and IgG were used to assay for rheumatoid factor (Rh-F) auto-antibodies. Generally, antibodies to parasite antigens in onchocerciasis patients were remarkably higher than control group (p&lt;0.05) using exact F-test. There was no significant difference (p&gt;0.05) in antibodies to nerve-DOC and Rh-F in patients compared to control. Antibodies increased with increasing skin snip microfilaria load from 0.69±0.28 with 0mf/mg (n=54) as against 0.80±0.26 for those with 4-20mf/mg. Observed slight negative correlation in IgG antibody levels and severity of disc colour with mean OD values of 0.26±0.22 in those graded as having no optic nerve disease (OND) (disc 1, n=86) and 0.17±0.19 for those with severe changes (disc 3, n=49) was not statistically significant (P&gt;0.05). An age dependent significant decrease (P&lt;0.05) in antibodies were observed with 0.64±0.34 for 15-30yr old (n=48) compared to 0.48±0.35 for those 50yr (n=50) for PBS with a similar trend for IgG to Ov1.9 and Cam1. In conclusion, serum parasite-specific and non-parasite antibodies may not be responsible for the pathology of optic nerve disease. Onchocerciasis patients were apparently not at higher risk of developing rheumatoid arthritis than the control.Keywords: Onchocerciasis; Antibodies; Antigens; Immune responses; Ivermectin.

Evaluation of Lymphatic Filariasis and Onchocerciasis in Three Senegalese Districts Treated for Onchocerciasis with Ivermectin.

In Africa, onchocerciasis and lymphatic filariasis (LF) are co-endemic in many areas. Current efforts to eliminate both diseases are through ivermectin-based mass drug administration (MDA). Years of ivermectin distribution for onchocerciasis may have interrupted LF transmission in certain areas. The Kédougou region, Senegal, is co-endemic for LF and onchocerciasis. Though MDA for onchocerciasis started in 1988, in 2014 albendazole had not yet been added for LF. The objective of this study was to assess in an integrated manner the LF and onchocerciasis status in the three districts of the Kédougou region after ≥10 years of ivermectin-based MDA. The study employed an African Programme for Onchocerciasis Control (APOC) onchocerciasis-related methodology. In the three districts, 14 villages close to three rivers that have Simulium damnosum breeding sites were surveyed. Convenience sampling of residents ≥5 years old was performed. Assessment for LF antigenemia by immunochromatographic testing (ICT) was added to skin snip microscopy for onchocerciasis. Participants were also tested for antibodies against Wb123 (LF) and Ov16 (onchocerciasis) antigens. In two districts, no participants were ICT or skin snip positive. In the third district, 3.5% were ICT positive and 0.7% were skin snip positive. In all the three districts, Wb123 prevalence was 0.6%. Overall, Ov16 prevalence was 6.9%. Ov16 prevalence among children 5–9 years old in the study was 2.5%. LF antigenemia prevalence was still above treatment threshold in one district despite ≥10 years of ivermectin-based MDA. The presence of Ov16 positive children suggested recent transmission of Onchocerca volvulus. This study showed the feasibility of integrated evaluation of onchocerciasis and LF but development of integrated robust methods for assessing transmission of both LF and onchocerciasis are needed to determine where MDA can be stopped safely in co-endemic areas.

An isothermal DNA amplification method for detection of Onchocerca volvulus infection in skin biopsies.

BackgroundDiagnostic procedures for the diagnosis of infection with the nematode parasite Onchocerca volvulus are currently based on the microscopic detection of microfilariae in skin biopsies. Alternative approaches based on amplification of parasitic DNA in these skin biopsies are currently being explored. Mostly this is based on the detection of the O-150 repeat sequence using PCR based techniques.MethodsAn isothermal, loop-mediated amplification method has been designed using the mitochondrial O. volvulus cox1 gene as a target.ResultsAnalysis of dilution series of synthetic DNA containing the targeted sequence show a non-linear dose-response curve, as is usually the case for isothermal amplification methods. Evaluation of cross-reactivity with the heterologous sequence from the closely related parasites Wuchereria bancrofti, Loa loa and Brugia malayi demonstrated strong specificity, as none of these sequences was amplified. The assay however amplified both O. volvulus and O. ochengi DNA, but with a different melting point that can be used to discriminate between the species. Evaluation of this assay in a set of skin snip biopsies collected in an endemic area in Ghana showed a high correlation with O-150 qPCR and also demonstrated a similar sensitivity. Compared to qPCR, LAMP had a sensitivity of 88.2% and a specificity of 99.2%.ConclusionsWe have developed a sensitive and specific loop-mediated amplification method for detection of O. volvulus DNA in skin biopsies that is capable of providing results within 30 min.Electronic supplementary materialThe online version of this article (doi:10.1186/s13071-016-1913-7) contains supplementary material, which is available to authorized users.

Public Health Analysis of Manifestation of Onchocerciasis in Rural Nigeria

Aspects of human infection with Onchocerca volvulus was investigated in 9 villages in Okigwe, Nigeria. A cross sectional survey method was adopted for the study, blood free skin snips were collected from randomly selected consenting adults aged 5–62 years also palpation for mobile subcutaneous lumps and clinical manifestation were observed among 960 persons comprising of 511 males and 449 females.

Evaluation of Onchocerciasis among community members of Okigwe Local Government Area, Imo State, Nigeria

Human infection with Onchocerca volvulus study was conducted to compare the prevalence of different manifestations of onchocerciasis according to gender and age, with view to determine if there had been a change in prevalence of onchocerciasis among residents of some communities in Okigwe Local Government Area of Imo State. A cross sectional survey method was adopted for the study. Skin snips were collected from randomly selected consenting adults aged 18-62 years. Palpation for mobile subcutaneous lumps and clinical manifestation were carried out on 960 persons comprising of 511 males and 449 females. The commonest symptoms observed were poor vision (8%), nodules (7%), leopard skin (9%), lizard skin (7%), and hanging groin (1%). Musculo-skeletal pain (MSP) was recorded as one of the major complaints by (14%) of the subjects. This result indicates (69.8%) reduction on clinical features of onchocerciasis prevalence among sampled population in the study-area.Keywords: onchocerciasis, manifestation, prevalence, palpation

Paramyosin of canine Onchocerca lupi: usefulness for the diagnosis of a neglected zoonotic disease.

BackgroundOf increasing importance to the medical and veterinary communities is the zoonotic filarioid nematode Onchocerca lupi. Onchocercosis, thus far found in wolves, dogs, cats and humans, is diagnosed via skin snips to detect microfilariae and surgical removal of adults from the eye of the host. These methods are time-consuming, laborious and invasive, highlighting the need for new tools for the diagnosis of O. lupi in susceptible hosts. Symptoms related to the presence of the adults in the eye can range from none apparent to severe, including blindness. No reliable chemotherapeutic protocols are available, as yet, to eliminate the infection. Paramyosin, an invertebrate-specific protein, has been well-studied as an allergen, diagnostic marker and vaccine candidate. The aim of this study, therefore, was to isolate and characterise paramyosin from O. lupi to assess its suitability for the development of a serological diagnostic assay.MethodsThe adult and microfilarial stages of O. lupi were isolated from the eyes and skin of a 3-year-old male dog. Total RNA was extracted and reverse transcribed into single stranded cDNA. Reverse-transcription PCR was used to isolate a full-length paramyosin cDNA from adult worms and to investigate the temporal expression patterns of this gene. All amplicons were sequenced using dideoxy chain termination sequencing. Bioinformatics was used to predict the amino acid sequence of the gene, to compare the DNA and protein sequences with those available in public databases and to investigate the phylogenetic relationship of all molecules. Antibody binding sites were predicted using bioinformatics and mapped along with published antigenic epitopes against the O. lupi paramyosin protein. The native protein, and three smaller recombinantly expressed peptides, were subjected to western blot using serum from dogs both positive and negative for O. lupi.ResultsParamyosin of O. lupi was herein molecularly characterized, encoded by a transcript of 2,643 bp and producing a protein of 881 amino acids (101.24 kDa). The paramyosin transcript was detected, by reverse transcription PCR, in adults and microfilariae, but not in eggs. Phylogenetic analysis indicates that this molecule clusters with paramyosins from other filarioids to the exclusion of those from other taxa. A total of 621 unique antibody binding epitopes were predicted for this protein and another 28 were conserved in other organisms. This information was used to design three peptides, for recombinant expression, to identify the antibody binding epitope(s) and reduce potential cross-reactivity with serum from dogs infected with other filarioid nematodes. Native paramyosin, purified from microfilariae and adults, was detected by antibodies present in serum from dogs with known O. lupi infections.ConclusionsData provided herein may assist in the development of a serological diagnostic test, based on antibodies to O. lupi paramyosin, for the diagnosis of this infection, in order to gain more information on the real distribution of this little known filarioid of zoonotic concern.Electronic supplementary materialThe online version of this article (doi:10.1186/s13071-016-1783-z) contains supplementary material, which is available to authorized users.

Comparison of a new visual isothermal nucleic acid amplification test with PCR and skin snip analysis for diagnosis of onchocerciasis in humans

Accurate, simple and affordable diagnostics are needed to detect Onchocerca volvulus infection in humans. A newly developed colorimetric loop-mediated isothermal amplification (LAMP) assay was compared to PCR and skin snip analysis for diagnosis of onchocerciasis. The robustness and simplicity of the assay indicates that it may be a useful field tool for surveillance in endemic countries.

Evidence for Suppression of Onchocerciasis Transmission in Bioko Island, Equatorial Guinea.

Onchocerciasis or "river blindness" is a chronic parasitic neglected tropical disease which is endemic both in mainland and insular Equatorial Guinea. We aim to estimate the current epidemiological situation of onchocerciasis in Bioko Island after vector elimination in 2005 and more than sixteen years of Community Directed Treatment with Ivermectin (CDTI) by using molecular and serological approaches for onchocerciasis diagnosis. A community-based cross-sectional study was carried out in Bioko Island from mid-January to mid-February 2014. A total of 544 study participants were recruited. A complete dermatological examination was performed and three skin snips were performed in every participant for parasitological and molecular assessments. Blood spots were also taken for determination of Ov16 IgG4 antibodies trough an “in-house” ELISA assay. Overall, we found 15 out of 522 individuals suffering any onchocerciasis specific cutaneous lesions and 16 out of 528 (3.0%) with onchocercal nodules in the skin. Nodules were significantly associated with age, being more common in subjects older than 10 years than in younger people (3.9% vs. 0%, p = 0.029). Regarding the onchocerciasis laboratory assessment, no positive parasitological test for microfilaria detection was found in the skin snips. The calculated seroprevalence through IgG4 serology was 7.9%. No children less than 10 years old were found to be positive for this test. Only one case was positive for Onchocerca volvulus (O. volvulus) after skin PCR. The present study points out that the on-going mass ivermectin treatment has been effective in reducing the prevalence of onchocerciasis and corroborates the interruption of transmission in Bioko Island. To our knowledge, this is the first time that accurate information through molecular and serological techniques is generated to estimate the onchocerciasis prevalence in this zone. Sustained support from the national program and appropriate communication and health education strategies to reinforce participation in CDTI activities are essential to ensure progress towards onchocerciasis elimination in the country.

The First Confirmed Elimination of an Onchocerciasis Focus in Africa: Abu Hamed, Sudan.

Mass treatment with ivermectin for onchocerciasis was stopped in 2012 in Abu Hamed, an isolated focus on the River Nile in northern Sudan. A 3-year posttreatment surveillance (PTS) ensued, at the end of which an evaluation was conducted in 2015 following the current World Health Organization guidelines for verification of onchocerciasis elimination. Vector black flies were collected from sentinel breeding sites and finger-prick bloodspots were collected from children ≤ 10 years of age resident in 35 communities within the focus. Polymerase chain reaction (PCR) screening of 19,191 flies from four sites for the O-150 parasite-specific marker found no flies carrying Onchocerca volvulus larvae (0%, 95% upper confidence limit [UCL] = 0.16), and serological testing of 5,266 children identified only one Ov16 seropositive child (0.019%, 95% UCL = 0.074); whose skin snips were negative when tested by O-150 PCR assay. These results indicate that for the first time in Africa, onchocerciasis elimination has been verified after a successful PTS in Abu Hamed.

Progress towards onchocerciasis elimination in the participating countries of the African Programme for Onchocerciasis Control: epidemiological evaluation results.

BackgroundThe African Programme for Onchocerciasis Control (APOC) was created in 1995 to establish community-directed treatment with ivermectin (CDTi) in order to control onchocerciasis as a public health problem in 20 African countries that had 80 % of the global disease burden. When research showed that CDTi may ultimately eliminate onchocerciasis infection, APOC was given in 2008 the additional objective to determine when and where treatment can be safely stopped. We report the results of epidemiological evaluations undertaken from 2008 to 2014 to assess progress towards elimination in CDTi areas with ≥6 years treatment.MethodsSkin snip surveys were undertaken in samples of first-line villages to determine the prevalence of O. volvulus microfilariae. There were two evaluation phases. The decline in prevalence was evaluated in phase 1A. Observed and model-predicted prevalences were compared after correcting for endemicity level and treatment coverage. Bayesian statistics and Monte Carlo simulation were used to classify the decline in prevalence as faster than predicted, on track or delayed. Where the prevalence approached elimination levels, phase 1B was launched to determine if treatment could be safely stopped. Village sampling was extended to the whole CDTi area. Survey data were analysed within a Bayesian framework to determine if stopping criteria (overall prevalence <1.4 % and maximum stratum prevalence <5 %) were met.ResultsIn phase 1A 127 665 people from 639 villages in 54 areas were examined. The prevalence had fallen dramatically. The decline in prevalence was faster than predicted in 23 areas, on track in another 23 and delayed in eight areas. In phase 1B 108 636 people in 392 villages were examined in 22 areas of which 13 met the epidemiological criteria for stopping treatment. Overall, 32 areas (25.4 million people) had reached or were close to elimination, 18 areas (17.4 million) were on track but required more years treatment, and in eight areas (10.4 million) progress was unsatisfactory.ConclusionsOnchocerciasis has been largely controlled as a public health problem. Great progress has been made towards elimination which already appears to have been achieved for millions of people. For most APOC countries, nationwide onchocerciasis elimination is within reach.Electronic supplementary materialThe online version of this article (doi:10.1186/s40249-016-0160-7) contains supplementary material, which is available to authorized users.

Filarioids infecting dogs in northeastern Brazil

Species of filarial nematodes belonging to the genera Dirofilaria and Acanthocheilonema are recognised as common parasites of dogs throughout the world. Recently, other filarioids featured by the presence of dermal microfilariae (e.g., Onchocerca lupi and Cercopithifilaria spp.) have been recognised in Europe. In Brazil, reports of filarioids in dogs are limited to Dirofilaria immitis, Acanthocheilonema reconditum and Cercopithifilaria bainae. To investigate the distribution of filarial infections in dogs living in an endemic region from northeastern Brazil, blood and skin samples (n=104) were microscopically (modified Knott’s test and skin snip sediment examination) and molecularly evaluated. Twenty-two dogs (21.15%) were positive at microscopic and/or molecular examination for at least one filarioid species, with 21 (20.19%) animals positive for blood microfilariae at molecular and/or at microscopic examination. Microfilariae of D. immitis were detected in 12 (11.54%) animals, with co-infection of D. immitis and A. reconditum observed in four (3.85%) individuals. One animal was positive for C. bainae at both microscopic and molecular examination. Analysis of sequence obtained in the present study showed significant alignment identity with that of C. bainae from Europe. Considering that in the area of study arthropod vectors (mosquitoes, fleas and ticks) are prevalent throughout the year, preventive measures should be disposed in order to avoid the animal infestation and pathogen infection.