Sensitivity of diagnostic methods for Mansonella ozzardi microfilariae detection in the Brazilian Amazon Region

BACKGROUND The human filarial worm Mansonella ozzardi is highly endemic in the large tributaries of the Amazon River. This infection is still highly neglected and can be falsely negative when microfilariae levels are low. OBJECTIVES This study investigated the frequency of individuals with M. ozzardi in riverine communities in Coari municipality, Brazilian Amazon. METHODS Different diagnostic methods including polymerase chain reaction (PCR), blood polycarbonate membrane filtration (PCMF), Knott's method (Knott), digital thick blood smears (DTBS) and venous thick blood smears (VTBS) were used to compare sensitivity and specificity among the methods. Data were analysed using PCMF and Bayesian latent class models (BLCM) as the gold standard. We used BLCM to calculate the prevalence of mansonelliasis based on the results of five diagnostic methods. FINDINGS The prevalence of mansonelliasis was 35.4% by PCMF and 30.1% by BLCM. PCR and Knott methods both possessed high sensitivity. Sensitivity relative to PCMF was 98.5% [95% confidence interval (CI): 92.0 - 99.7] for PCR and 83.5% (95% CI: 72.9 - 90.5) for Knott. Sensitivity derived by BLCM was 100% (95% CI 93.7 - 100) for PCMF, 100% (95% CI: 93.7 - 100) for PCR and 98.3% (95% CI: 90.6 - 99.9) for Knott. The odds ratio of being diagnosed as microfilaremic increased with age but did not differ between genders. Microfilariae loads were higher in subjects aged 30 - 45 and 45 - 60 years. MAIN CONCLUSIONS PCMF and PCR were the best methods to assess the prevalence of mansonelliasis in our samples. As such, using these methods could lead to higher prevalence of mansonelliasis in this region than the most commonly used method (i.e., thick blood smears).

Mansonella ozzardi (Nematoda: Onchocercidae) in the riverine population of the Tefé River, State of Amazonia, Brazil

Introduction: This study assessed the prevalence of Mansonella ozzardi in riverine communities of the Tefé River, Amazonas, Brazil. Methods: The prevalence of M. ozzardi was estimated by microscopic examination of thick blood smears. Results: The M. ozzardi prevalence rate was 6.3% (19/300). Filarial infection was found in 8 of the 11 communities surveyed, with prevalence rates varying from 2.5% to 22.2%. Conclusions: Tefé is a region of oil and natural gas exploration, in which there is a high turnover of workers. Migration patterns may facilitate the dissemination of mansonelliasis to other regions. .

Reporte de cuatro casos clínicos de filariasis en Alto Nanay, Loreto / Report of four clinical cases of filariasis in Alto Nanay, Loreto

El presente estudio describe los hallazgos clínicos, parasitológicos y de laboratorio de cuatro pacientes residentes en el distrito de Alto Nanay, Maynas, Loreto, Perú; infectados con microfilarias de Mansonella ozzardi. La evaluación clínica incluyó las especialidades de oftalmología, cardiología y nefrología. En el caso 1, destaca la presencia de dos tumoraciones subcutáneas, una a nivel dorsal y la otra en el tercio inferior de la pierna izquierda; en el caso 2, la sensación de “hormigueo o frío en las piernas”; en el caso 3 se encontró asociada una infección crónica por hepatitis B y en el caso 4 una tumoración de gran tamaño en región lumbar izquierda. En los cuatro pacientes se encontró Mansonella ozzardi y eosinofilia, tres presentaron valores mayores a 20%. La tumoración en el caso 4 correspondió a una hernia de contenido intestinal. Es necesario ejecutar más estudios clínicos y evaluar su verdadero efecto patógeno, es también pertinente estudiar la diversidad genética de filarias de la Amazonia peruana.

This study describes the clinical, parasitological and laboratory findings of four patients who lived in the district of Alto Nanay, Maynas, Loreto, Peru and were infected with Mansonella ozzardi microfilariae. Clinical examinations by ophthalmologists, cardiologists and nephrologists were performed. In case 1, the presence of 2 subcutaneous lumps was the most important finding, one at dorsal level and the other in the lower third of the left leg; in case 2, there was a sensation of tingling or coldness in the legs; in case 3, an associated chronic hepatitis B infection was found, and in case 4, a large lump was detected in the left lumbar region. All 4 patients were infected with Mansonella ozzardi and had eosinophilia, 3 of them in percentages of 20%. The lump found in case 4 was due to a herniation of bowel content. Further clinical studies and an evaluation of the actual pathogenic effect of microfilariae are to be performed. A study of the genetic diversity of filariae in the Peruvian Amazon would also be important.

Aporte de la técnica de PCR en el diagnóstico de Mansonella ozzardi en zonas endémicas de la Argentina / Contribution of the PCR assay to the diagnosis of Mansonella ozzardi in endemic areas of Argentina

Mansonella ozzardi es un nematode parásito tisular, agente etiológico de mansonellosis en casi la totalidad de los países latinoamericanos. En Argentina la mansonellosis ha sido descrita a lo largo de la región de las yungas. Su diagnóstico microscópico puede dar resultados falsos negativos en microfilaremias bajas. El objetivo del presente estudio fue optimizar su diagnóstico molecular y comparar los resultados con los obtenidos mediante las pruebas microscópicas de Knott, de gota gruesa y de extendido hemático fino, en 92 muestras de sangre de pacientes de zona endémica. La técnica de PCR seguida de la secuenciación del producto amplificado presentó una sensibilidad del 100 % frente al método de Knott, considerado como referencia, e incluso permitió identificar 7 casos más de la parasitosis.

Mansonella ozzardi is a tissue-dwelling parasitic nematode, the causative agent of mansonelliasis in almost all Latin American countries. It has been described along the Argentine Yungas region. The microscopic diagnosis can yield false-negative test results at low microfilaremia levels. The aim of this study was to optimize the molecular diagnostic technique and compare it with the Knott's method and standard blood smear procedures (thin blood films and thick smears) in 92 blood samples of individuals from an endemic area. The PCR technique followed by the sequencing of the amplified product yielded 100 % sensitivity compared to the Knott's test, which is considered a reference method. Seven more cases of this parasitosis could only be identified with the molecular technique.

Investigation of the occurrence of Mansonella ozzardi in the State of Rondônia, Western Amazonia, Brazil / Investigação da ocorrência da Mansonella ozzardi no Estado de Rondônia, Amazônia Ocidental

INTRODUCTION: Mansonella ozzardi, a human filarial nematode transmitted by Diptera Simuliidae flies (black flies), is widely spread in the Amazon, while the occurrence of mansonelliasis in the State of Rondônia (Western Amazonia) has not been reported thus far. The existence of this parasitic disease in this state is investigated in this study. METHODS: A total of 4,452 people who lived alongside the Madeira, Mamore, Guapore, Machado and Preto Rivers, in the State of Rondônia, were examined using the thick blood smear method. Alongside the Machado and Guapore rivers, 2,112 adult simuliid specimens were captured. RESULTS: Infected people were not reported, although simuliids with vector potential such as Cerqueirellum pydanielli, C. argentiscutum and C. amazonicum were found. CONCLUSIONS: This study registered no infected people. This finding may be explained by the individuals' low level of microfilaremia that is below the limit of detection of the diagnostic method used in the study or insufficient migration of people infected with M. ozzardi from endemic areas, making it impossible to introduce the disease into Rondônia. Further studies using more sensitive diagnostic techniques could shed light on this question.

INTRODUÇÃO: Mansonella ozzardi é uma filária humana que tem como vetor dípteros simulídeos e é amplamente distribuída na Amazônia. Não há informações sobre a ocorrência de casos de mansonelose no Estado de Rondônia, e neste trabalho procurou-se investigar a situação epidemiológica desta parasitose em áreas urbanas e ribeirinhas na Amazônia Ocidental. MÉTODOS: Foram examinados 4.452 moradores às margens dos rios Madeira, Mamoré, Guaporé, Machado e Preto em Rondônia, através da técnica da gota espessa de sangue. Capturou-se às margens do rio Machado e Guaporé 2.112 simulídeos adultos das espécies Cerqueirellum pydanieli e Chirostilbia pertinax. RESULTADOS: Não foram encontradas pessoas infectadas com M. ozzardi e foram encontradas diferentes espécies de simulídeos com potencial de transmissão da mansonelose. CONCLUSÕES: Não foram encontrados indivíduos infectados com M. ozzardi neste estudo. Este fato pode ser explicado pela existência de pacientes com baixas microfilaremias, associado à técnica diagnóstica de baixa sensibilidade utilizada no estudo. Pode-se ainda aventar a possibilidade de ter ocorrido migração insuficiente de pacientes infectados de áreas endêmicas, impossibilitando a implantação da endemia. Estudos posteriores com técnicas diagnósticas mais sensíveis podem ajudar a esclarecer esta questão.

Improvement of a PCR test to diagnose infection by Mansonella ozzardi / Adequação da técnica da PCR para diagnóstico de infecção de Mansonella ozzardi

INTRODUCTION: Mansonelliasis is caused by Mansonella ozzardi. It is widespread in the Amazon region, with a high prevalence. The common exam of thick blood smears stained with Giemsa shows low efficacy levels and has been an obstacle to diagnosing individuals with low blood parasitemia. METHODS: In order to increase diagnosis efficacy, the PCR technique was improved. RESULTS AND CONCLUSIONS: PCR demonstrated the best performance, with sensitivity and negative predictive values (NPV) of 100 percent, followed by blood filtration through membrane filters, which showed a sensitivity of 88.9 percent and a NPV of 84.6 percent, when compared to thick blood smears.

INTRODUÇÃO: A mansonelose é uma filariose causada pela Mansonella ozzardi, ocorrendo na Amazônia com prevalências de até 60 por cento. A técnica de diagnóstico habitual (hemoscopia através da gota espessa) tem baixa eficácia o para o diagnóstico de pacientes com baixa parasitemia. MÉTODOS: Neste contexto foi aperfeiçoada a técnica da PCR para seu diagnóstico. RESULTADOS E CONCLUSÕES: Quando comparada à gota espessa, a PCR apresenta sensibilidade de 100 por cento, e valor preditivo negativo (VPN) de 100 por cento mostrando eficácia bastante superior à técnica da filtração em membrana que apresenta sensibilidade de 88,9 por cento e VPN de 84,6 por cento, quando também comparada à gota espessa de sangue.

Prevalence of Mansonella ozzardi among riverine communities in the municipality of Lábrea, State of Amazonas, Brazil / Prevalências de Mansonella ozzardi entre comunidades ribeirinhas do município de Lábrea, Estado do Amazonas

INTRODUCTION: Estimate the prevalence of Mansonella ozzardi infection and calculate the parasitic infection rate (PIR) in simuliid black flies in the municipality of Lábrea, State of Amazonas, Brazil. METHODS: Prevalence was measured using the thick blood smear method collected from the fingers and was related to age, sex and occupation. Simuliidae were collected with a suction apparatus, then stained with hematoxylin and dissected to verify the PIR. RESULTS: The average prevalence rate of M. ozzardi among the 694 individuals examined was 20.7 percent. Infection was higher in men (27.6 percent) than in women (14.3 percent) (p < 0.001) and occurred in most age groups, with the highest prevalence in the following age groups: 38-47 (40 percent), 48-57 (53.1 percent) and >58 (60.5 percent). The highest prevalence rates were observed in the retired (64 percent), followed by farm workers (47.1 percent). Infection by M. ozzardi was only identified in Cerqueirellum amazonicum (Simuliidae) with a PIR of 0.6 percent. CONCLUSIONS: This study showed a high prevalence of M. ozzardi in the riverine communities of Lábrea due to the lack of policies regarding the treatment of microfilaremic individuals in the region and an abundance of competent vectors for M. ozzardi.

INTRODUÇÃO: Estimar as prevalências de Mansonella ozzardi, e calcular a taxa de infecção parasitária nos simulídeos no município de Lábrea, Estado do Amazonas. MÉTODOS: As prevalências foram obtidas através do método de gota espessa a partir do sangue coletado da polpa digital e foram relacionadas com a idade, sexo e ocupação. Os simulídeos foram coletados com capturador de sucção, posteriormente foram corados com hematoxilina e dissecados para se obter a taxa de infecção parasitária. RESULTADOS: A prevalência de M. ozzardi entre as 694 pessoas examinadas foi de 20,7 por cento. A infecção foi maior nos homens (27,6 por cento) do que nas mulheres (14,3 por cento) (p < 0,001) e ocorreu na maioria das faixas etárias com maiores prevalências nos indivíduos entre 38-47 (40 por cento), 48-57 (53,1 por cento) e >58 (60,5 por cento) anos. As maiores prevalências foram observadas nos aposentados (64 por cento) e agricultores (47,1 por cento). Somente Cerqueirellum amazonicum (Simuliidae) foi encontrado infectado com M. ozzardi com uma taxa de infecção parasitária de 0,6 por cento. CONCLUSÕES: Este trabalho mostra elevadas prevalências de M. ozzardi nas comunidades ribeirinhas de Lábrea devido à ausência de políticas de tratamento dos microfilarêmicos na região e a abundância de vetores competentes.

Mansonella ozzardi in Brazil: prevalence of infection in riverine communities in the Purus region, in the state of Amazonas

This study was undertaken to investigate the prevalence of Mansonella ozzardi infection and to estimate the parasitic infection rate (PIR) in simuliid black flies in the municipality of Pauini, Amazonas, Brazil. We used thick blood films to examine 921 individuals in 35 riverine communities along the Pauini and Purus Rivers. Simuliids were caught in several communities. Flies were identified, stained with haematoxylin and dissected. Overall, 44 (24.86 percent) of 177 riverines were infected in communities on the Pauini River and 183 (24.19 percent) of 744 on the Purus. The prevalence was higher in men (31.81 percent and 29.82 percent) than in women (17.98 percent and 19.18 percent) and occurred in most age groups. The prevalence increased sharply in the 28-37 (50 percent and 42.68 percent) age group and increased in the older age classes. The highest prevalence was in farmers (44 percent and 52.17 percent, respectively) in the Pauini and Purus Rivers. Only Cerqueirellum amazonicum (Simuliidae) transmits M. ozzardi in this municipality, and we found a PIR of 0-8.43 percent and infectivity rate of 0-3.61 percent. These results confirm that rates of M. ozzardi infection are high in Pauini and suggest that its prevalence may be far greater than has been previously reported due to the absence of a program for treating the population.

Current profile of Mansonella ozzardi (Nematoda: Onchocercidae) in communities along the Ituxi river, Lábrea municipality, Amazonas, Brazil

The objective of this paper was to describe the current status of Mansonella ozzardi prevalence among the inhabitants of 12 communities along the Ituxi river, in Lábrea municipality, state of Amazonas. The prevalence of M. ozzardi was determined using thick blood smears obtained by digital punctures. M. ozzardi was found in 30.23 percent of the samples collected (39/129), with similar prevalence between genders (males: 30.30 percent; females: 30.16 percent); the highest prevalence was found in homemakers (45.45 percent) followed by farmers (38.77 percent). Among age groups, males and females older than 48 exhibited the highest rates. These results show a significative increase in the prevalence when compared to a epidemiological study made 26 years ago in the same area as well as a different epidemiological profile (gender and occupation) in relation to other areas in Amazonas.

Comparación de pruebas de diagnóstico de mansonelosis en el estudio de prevalencia en comunidades del río Atabapo, departamento del Guainía, Colombia 1996 / Comparison of mansonellosis diagnostic test in a prevalency study in communities situated along the River Atabapo in the Guainía Department, Colombia, 1996

En el departamento de Guainía, Colombia, se realizó un estudio tipo encuesta de prevalencia de corte en siete comunidades del río Atabapo, donde se calculó una muestra poblacional de 181 personas mayores de 10 años que se debían examinar, tomando, como unidad de análisis, 37 viviendas escogidas aleatoriamente. Se les tomó una muestra de sangre que se examinó mediante cuatro técnicas (examen en fresco, gota gruesa, prueba de Knott y conteo por cm cúbicos), examen médico, estudio de saneamiento y encuesta por vivienda. Se encontró una prevalencia de 57 por ciento en el grupo examinado que, por ser muestra representativa de la población, se puede decir que es la prevalencia de la zona. De las tres pruebas diagnósticas utilizadas, además de la prueba de knott, el conteo por cm cúbicos fue la que mejor resultado brindó en el estudio, dando una sensibilidad, especificidad y valor predictivo positivo similar y mejor que la de la prueba de oro que se tomó como referencia

Mansonelosis en el area Sur-oriental de la Orinoquia Venezolana / Mansonelliasis in the southeast Venezuelan Orinoquia region

Physical exam and skin biopsy were performed and peripheral blood samples drawn from 10 of the inhabitants of 26 jungle villages of the State of Bolívar, Venezuela. One hundred and fifty three Indians and an inhabitant of mixed blood from 13 different communities were found to be infected with mansonelliasis representing a global index of 36.40 in the endemic area. The parasitosis was found to be concentrated in three areas. The largest, located in the southwest of the country and forming part of an extent infected area embarking neighboring parts of Venezuela and Brazil, contains villages with a parasite index of 80.76 and 94.44. A second focus is located in the southeast, its carriers, as those of the first, are infected with Mansonella ozzardi. In a third, central eastern focus infections with Mansonella pertans were found. The patients infected with M. ozzardi were apparently asymptomatic and their physical exam was normal, even though individual parasite densities up to 30,000 microfilariae/ml blood and 22,000/gr skin were found.