Detection of DNA of filariae closely related to Mansonella perstans in faecal samples from wild non-human primates from Cameroon and Gabon.

BACKGROUND: The Onchocercidae is a family of filarial nematodes with several species of medical or veterinary importance. Microfilariae are found in the blood and/or the dermis and are usually diagnosed in humans by microscopy examination of a blood sample or skin biopsy. The main objectives of this study were to evaluate whether filariae DNA can be detected in faecal samples of wild non-human primates (NHPs), whether the detected parasites were closely related to those infecting humans and whether filarial DNA detection in faeces is associated with co-infections with nematodes (Oesophagostumum sp. and Necator sp.) known to cause blood loss while feeding on the host intestinal mucosa. METHODS: A total of 315 faecal samples from 6 species of NHPs from Cameroon and Gabon were analysed. PCRs targeted DNA fragments of cox1 and 12S rDNA genes, to detect the presence of filariae, and the internal transcribed spacer 2 (ITS2), to detect the presence of Oesophagostomum sp. and Necator sp. infections. RESULTS: Among the 315 samples analysed, 121 produced sequences with > 90% homology with Onchocercidae reference sequences. However, 63% of the 12S rDNA and 78% of the cox1 gene sequences were exploitable for phylogenetic analyses and the amplification of the 12S rDNA gene showed less discriminating power than the amplification of the cox1 fragment. Phylogenetic analyses showed that the cox1 sequences obtained from five chimpanzee DNA faecal samples from Gabon and two from Cameroon cluster together with Mansonella perstans with high bootstrap support. Most of the remaining sequences clustered together within the genus Mansonella, but the species could not be resolved. Among the NHP species investigated, a significant association between filarial DNA detection and Oesophagostomum sp. and Necator sp. infection was observed only in gorillas. CONCLUSIONS: To our knowledge, this is the first study reporting DNA from Mansonella spp. in faecal samples. Our results raise questions about the diversity and abundance of these parasites in wildlife, their role as sylvatic reservoirs and their potential for zoonotic transmission. Future studies should focus on detecting variants circulating in both human and NHPs, and improve the molecular information to resolve or support taxonomy classification based on morphological descriptions.

In vitro maintenance of Mansonella perstans microfilariae and its relevance for drug screening.

BACKGROUND: Mansonellosis arises from infections with threadlike filarial nematodes in millions of individuals, especially in sub-Saharan Africa. Since infections present no overt clinical symptoms but attenuate immune responses that might lead to increased susceptibility and worsened disease course of concomitant infections, it is truly a neglected tropical disease. Nevertheless, only few studies focus on identifying suitable safe drugs for its control and little is known about the requirements for in vitro maintenance of the Mansonella perstans transmission stage. This study, therefore, evaluated the survival of M. perstans microfilariae (mf) using in vitro conditions that have been shown to promote survival of Loa loa, a closely related filarial nematode. Furthermore, the in vitro microfilaricidal effect of 15 agents was assessed on this helminth. METHODS: The ability of two basic culture media; Dulbecco's Modified Eagle's Medium (DMEM) and Roswell Park Memorial Institute (RPMI-1640) supplemented with 10% fetal bovine serum (FBS) and a monkey kidney epithelial cell line (LLC-MK2) to support the survival of M. perstans microfilariae was investigated. Subsequently, 6 anti-helminthics, 5 anti-malarials, 1 anti-microbacterial, 2 trypanocidals and 1 anti-cancer agent were tested in vitro against mf. The suitability of the culture media as well as the effect of the anti-infective agents on mf survival was assessed by scoring their motility. RESULTS: FBS supplement and additional LLC-MK2 cells significantly improved the survival of mf in DMEM and RPMI-1640 culture. In detail, RPMI-1640 supplemented with 10% FBS and LLC-MK2 cells sustained the maintenance of mf for at least 20 days (100.00 ±â€¯0.00% survival). In co-cultures with LLC-MK2 cells without serum, M. perstans mf were maintained in DMEM and RPMI-1640 medium with a motility above 99% by day 5. Mefloquine displayed the highest microfilaricidal effect in vitro followed by artesunate. CONCLUSION: Both RPMI and DMEM in the presence of LLC-MK2 cells are suitable for the maintenance of M. perstans mf in vitro. In absence of the feeder cells, the addition of 10% FBS to RPMI-1640 medium improved the parasite survival rate and motility. The microfilaricidal activity of mefloquine and artesunate on M. perstans mf was documented for the first time in this study and can therefore be considered as reference for further screening of agents against this parasite stage.

First report on surface aspects of Mansonella ozzardi (spirurida: onchocercidae) microfilariae by scanning electron microscopy: preliminary results

Blood samples from Mansonella ozzardi infected volunteers from Vila Antimary (Amazonas State) were processed and a protocol to isolate and prepare microfilariae was carried out in order to perform Scanning Electron Microscopy (SEM) analysis. Data obtained from ultrastructure showed some undescribed structural points of the parasite such as a dimple in the anterior end of the larva and small points ­orifice-like­ that may be related to amphidial structures or simply pores. Another interesting feature was the tip of the tail which is very similar to that found in the rodent parasite Dunnifilaria meningica

Hallazgo incidental de microfilarias de Mansonella ozzardi en un paciente pediátrico con linfoma T anaplásico / Incidental finding of Mansonella ozzardi in a paediatric patient with anaplastic lymphoma

Resumen Las filariasis son parasitosis producidas por nemátodos hemáticos de la familia Filariidae, la Mansonella ozzardi, es uno de los agentes etiológicos distribuido ampliamente en el continente americano y en el Caribe. Presentamos el caso de una paciente de 13 años de edad, previamente diagnosticada con linfoma T de célula grande anaplásico. Como parte de la evaluación antes del segundo ciclo B de quimioterapia, se realizó un extendido de sangre periférica en el que se encontró una microfilaría tipificada como Mansonella ozzardi, se dio manejo con una dosis única de ivermectina y se logró resultado negativo en el control a las 24 horas. Actualmente la paciente se encuentra asintomática y sin evidencia de recurrencia de la parasitemia y terminando su tratamiento oncológico.

Abstract Filariasis is caused by nematodes in the blood. Mansonella ozzardi is one of the aetiological agents widely distributed in the Americas and the Caribbean. The case is presented on a paediatric patient previously diagnosed with T-cell anaplastic large cell lymphoma. As part of the evaluation before the second cycle B chemotherapy, a peripheral blood smear was performed, in which were found microfilaria, identified as Mansonella ozzardi. The treatment was a single dose of ivermectin, with a negative result being obtained at 24 hours. The patient is currently asymptomatic and with no evidence of recurrence of the parasitaemia, and able to finish the cancer treatment.

CD39 and immune regulation in a chronic helminth infection: The puzzling case of Mansonella ozzardi.

BACKGROUND: Chronic helminth infections typically induce an immunoregulatory environment, with markedly reduced immune responses to both parasite-specific and unrelated bystander antigens. Here we tested whether these changes are also observed in human infections with Mansonella ozzardi, a neglected filarial nematode widely distributed across Latin America. METHODS: CD4+ T cell populations from microfilaremic (Fil+) and uninfected (Fil-) inhabitants in M. ozzardi-endemic riverine communities in Brazil were characterized by flow cytometry analysis. Plasma concentrations of a wide range of cytokines and chemokines were measured. We examined whether M. ozzardi infection is associated with suppressed in vitro lymphoproliferative and inflammatory cytokine responses upon stimulation with filarial antigen, unrelated antigens or mitogens. PRINCIPAL FINDINGS/CONCLUSIONS: Fil+ subjects had lower plasma levels of selected inflammatory cytokines, such as TNF-α, IL-8, and IL-6, than their Fil- counterparts. However, we found no evidence for attenuated T-cell responses to filarial antigens or co-endemic pathogens, such as malaria parasites and Toxoplasma gondii. CD4+ T cells expressing CD39, an ectonucleosidase involved in the generation of the anti-inflammatory molecule adenosine, were increased in frequency in Fil+ subjects, compared to uninfected controls. Significantly, such an expansion was directly proportional to microfilarial loads. Surprisingly, CD39 blocking with a neutralizing antibody suppressed antigen-driven lymphoproliferation in vitro, while decreasing inflammatory cytokine responses, in Fil+ and Fil- individuals. These findings suggest that circulating CD4+ CD39+ T cells comprise subsets with both regulatory and stimulatory roles that contribute to the immune homeostasis in chronic M. ozzardi infection.

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 perstans microfilaremic individuals are characterized by enhanced type 2 helper T and regulatory T and B cell subsets and dampened systemic innate and adaptive immune responses.

The filarial nematode Mansonella perstans is endemic throughout Africa, northern South America and the Caribbean. Interestingly, M. perstans-infected individuals present no distinct clinical picture associated with certain pathology. Due to its relatively silent nature, research on this tropical disease has been neglected, especially M. perstans-driven immune responses. A hindrance in obtaining data on M. perstans-specific responses has been the inability to obtain adult worms since their habitats in serous cavities are difficult to access. Thus, in this study, for the first time, we used Mansonella perstans worm antigen extract as stimulant to obtain filarial-specific recall and immunoglobulin responses from M. perstans microfilaremic individuals (Mp MF+) from Cameroon. Moreover, systemic immune profiles in sera and immune cell composition in peripheral blood from Mp MF+ and amicrofilaremic individuals (Mp MF-) were obtained. Our data reveal that Mp MF+ individuals showed significantly reduced cytokine (IL-4, IL-6 and IL-12p70) and chemokine levels (IL-8 and RANTES), but significantly higher MIP-1ß as well as increased M. perstans-specific IgG4 levels compared to Mp MF- individuals. In contrast, upon re-stimulation with worm antigen extract, IFN-γ, IL-13, IL-10 and IL-17A secretion was enhanced in cell cultures from Mp MF+ individuals when compared to those from cultures of healthy European individuals. Moreover, analysis of immune cell composition in peripheral blood from Mp MF+ individuals revealed increased type 2 helper T (Th2), natural killer (NK), regulatory B and T cell (Breg and Treg) subsets but decreased type 1 regulatory T (Tr1) cells. In summary, this study deciphers for the first time, M. perstans-specific immune responses using worm antigen extract and shows that patent M. perstans infections have distinct Th2, Breg and Treg subsets accompanied with reduced systemic innate and adaptive immune responses and dominant filarial-specific IgG4 levels.

Immune response in Mansonella ozzardi infection modulated by IL-6/IL-10 axis in Amazon region of Brazil.

Mansonellosis is an endemic disease in the South and Central America. In Brazil, one of the etiological agents is Mansonella ozzardi. This filarial infection is yet poorly understood, with a controversial morbity, presenting since a oligosymptoms, malaria-like signs or without complaint in humans. The knowledge of the human immune response to microfilariae infection is limited mainly by different evolutionary cycles of the parasite in the host. In addition, the prevalence of this filarial parasite infection is high in several regions of Amazonas State. A cross-sectional study was conducted in an endemic area for microfilariae of M. ozzardi (MF) infection in the Amazonas State, Brazil. Proinflammatory and regulatory cytokines (IL-2, IL-4, IL-6, IL-10, TNF, IFN-gamma, and IL-17A) were measured in cryopreserved serum using the Cytometric Bead Array techniques (CBA) in 54 patients diagnosed with M. ozzardi infection and 55 individuals without the infection were included in the study (Controls). The IL-4, IL-6 and IL-10 level increased in infected patients with MF infection, while IL-17A increased in control only. When we compared controls to patients with high or low parasite load, the increased level of IL-6 and IL-10 were maintained. IL-6 contributes to the proinflammatory activity and IL-10 modulates Th1, Th2 and Th17 immune response. Furthermore, IL-4 was detected as a marker in the MF infection and MF patients with low parasite load, indicating the action of the Th2 cell response. The complex network of cytokines acting during M. ozzardi infection depends on a fine balance to determine a host protective effect or filarial persistence. Therefore, these results suggest that the immune response in MF infection is modulated by IL-6/IL-10 axis.

Successful long-term maintenance of Mansonella perstans in an in vitro culture system.

BACKGROUND: Approximately 114 million people are infected with Mansonella perstans in large proportions of Africa. In contrast to other filariae that infect humans, M. perstans-infected individuals show no distinct pathology or specific clinical picture, indicating a well-tuned adaptation to the host. In addition, since M. perstans adult worms reside in serous cavities which are difficult to access, research has been hindered and there is a paucity of knowledge about the biology of M. perstans, especially the development of the different life stages as well as M. perstans-driven immune responses. Thus in this study, an in vitro culture system was developed which allows an in-depth analysis of M. perstans. RESULTS: Culicoides species were caught in Ediki (Kumba), Southwest Region within Cameroon following a blood meal on a microfilaremic donor that had 1500 microfilariae/ml of peripheral blood and kept in captivity for 12 days at 23 °C. In a pilot experiment, 15 infective larvae were obtained from the midges and co-cultured with a confluent monolayer of monkey kidney epithelial cells (LLC-MK2) in DMEM medium supplemented with 10% FBS for up to 77 days. The resulting survival rates of 33% revealed that the cell-conditioned medium was suitable for long-term maintenance of M. perstans worms. To confirm these preliminary observations, 249 infective larvae were cultured for 50 days and their development was monitored daily and microscopically graded for motility. In total, 170 (68.3%) filariae survived and 124 (49.8%) larvae moulted between days 21-30 to become L5 stage larvae which were motile and showed continuous vigorous movement. CONCLUSION: We have established an in vitro culture system for the generation and long-term maintenance of viable M. perstans worms. This technique will be an important tool to study parasite biology and development, the role in host immunity, and might be helpful to discover novel treatment strategies against this filariae.

Dirofilaria repens microfilariae from a human node fine-needle aspirate: a case report.

BACKGROUND: Human dirofilariosis is still a little known infection even in endemic areas. Dirofilariosis is zoonotic infection usually abortive in humans; instead, we report a very rare case (the 4th in the world), the first in Italy, in which at least two infective larvae became mature adults that mated and produced active microfilariae even though they did not reach peripheral blood. CASE PRESENTATION: A 30-year-old Italian woman presented with a transient oedematous swelling on the left abdominal wall with a creeping eruption followed by the occurrence of a subcutaneous nodular painless mass in the iliac region. One month later, a similar temporary swelling appeared on the contralateral inguinal region associated with intermittent joint discomfort in both knees. The patient had recently travelled abroad, therefore many possible diagnoses were to be ruled out. Routine laboratory investigations revealed eosinophilia. An ultrasound examination of the iliac swelling evidenced a well-defined cyst with a big filamentous formation in continuous movement. A fine-needle aspiration of the lesion was performed for parasitological, cytological and histological exams. The prompt microscopic examination of the aspired material showed the presence of numerous microfilariae that were initially morphologically attributed to Mansonella ozzardi. Subsequently, the revision of the Giemsa stained film and molecular analyses of the biological material, allowed to identify Dirofilaria repens as etiological agent of infection. CONCLUSIONS: We report of a case in whom microfilariae were detected in fine-needle aspirate of subcutaneous node, without evidence of microfilaraemia, and the infection failed to become fully patent. Therefore we confirm that complete development and fertilization of D. repens worms in human hosts may occur, at variance with what is commonly believed, that Dirofilaria worms cannot fully develop in humans.

Prevalencia y factores asociados con la filariosis por Mansonella ozzardi en 2 comunidades periurbanas de Iquitos, 2009 / Prevalence of filariasis by Mansonella ozzardi and associated factors among 2 suburban communities of Iquitos, 2009

Introducción: Tres casos de infección por Mansonella ozzardi (M. ozzardi) en muestras de malaria que provenían de 2 comunidades periurbanas de Iquitos (Perú) dieron la sospecha de transmisión local. El objetivo del estudio fue determinar la prevalencia y factores asociados con la filariosis por M. ozzardi en estas comunidades. Materiales y métodos: Se realizó un estudio transversal en las comunidades de Laguna Azul y La Unión en Iquitos. A todos los pobladores se les tomó una gota gruesa para la detección de M. ozzardi y se les realizó una encuesta epidemiológica. Se realizó colecta de larvas y mosquitos para identificación de potenciales vectores. Se calcularon los OR ajustados (ORa) usando regresión logística múltiple. Resultados: Se evaluaron a 433 personas; 58% fueron mujeres; la mediana de edad fue de 20 años; 58,2% fueron migrantes, 3,7% pescadores y 12,9% habían viajado por las cuencas del Alto anay. La prevalencia de M. ozzardi fue de 1,4% (6/433) y el más joven de los casos tuvo 31 años; se encontró asociación con ser pescador (ORa: 8,7; IC 95: 1,1-76,0) y con haber realizado viajes por la cuenca del Alto Nanay (ORa: 11,2; IC 95: 1,2-112,5). No se evidenció densidad significativa de vectores simúlidos o culicoides. Conclusión: Las comunidades estudiadas tuvieron una baja prevalencia de M. ozzardi y por la edad de los casos y los factores asociados encontrados se sospecha infección foránea a la zona de estudio, en especial en la cuenca del Alto Nanay.

Introduction: Three cases of M. ozzardi infection from 2 suburban communities in Iquitos, Peru, led to a suspicion of local transmission. The objective of the study was to determine the prevalence of filariasis by M. ozzardi and its associated factors among these communities. Materials and methods: A cross-sectional study was performed, as well as an epidemiological survey and a thick smear blood test. Larvae and mosquitoes were collected. The adjusted OR (aOR) using multiple logistic regression was calculated. Results: A total of 433 participants were enrolled; 58% were women. The prevalence of M. ozzardi was 1.4% and the associated factors included being a fisherman (aOR: 8.7; 95%CI: 1.1-76.0) and being on the Alto Nanay river (aOR: 11.2; 95%CI: 1.2-112.5). No significant evidence of simulidos or culicoides was found. Conclusion: The low prevalence of M. ozzardi found suggests a foreign infection, probably derived from the Alto Nanay river.

Mansonelosis por Mansonella ozzardi en voluntarios para despistaje de malaria en la Amazon¡a peruana / Mansonelosis by Mansonella ozzardi in volunteers undergoing screening for malaria in the peruvian Amazon

Objetivos. Determinar la frecuencia, manifestaciones cl¡nicas y factores asociados a la infecci¢n por Mansonella ozzardi en voluntarios de una campa¤a de despistaje de malaria en el distrito de Alto Nanay en la selva amaz¢nica del Per£. Materiales y m‚todos. Se realiz¢ un estudio descriptivo, anal¡tico y transversal. Los participantes fueron entrevistados y examinados por un m‚dico y los datos fueron registrados en una ficha cl¡nica. El diagn¢stico de infecci¢n por M. ozzardi se realiz¢ mediante la t‚cnica de gota gruesa y frotis. Resultados. La frecuencia de mansonelosis en esta poblaci¢n fue 47,8% (IC 95%: 39,1–56,6). Mediante el an lisis bivariado se encontr¢ que el reporte de disminuci¢n de la agudeza visual o visi¢n borrosa y la presencia de tumoraciones subcut neas fueron los signos y s¡ntomas estad¡sticamente asociados con la infecci¢n por microfilarias (p<0,05). La regresi¢n log¡stica encontr¢ asociaci¢n estad¡stica para la residencia en localidades del r¡o Pintuyacu o Alto Nanay, la ocupaci¢n en lugares alejados del centro poblado, la presencia de tumores subcut neos y el engrosamiento de la piel (p<0,05).Conclusiones . Existe una elevada frecuencia de mansonelosis por M. ozzardi en el distrito de Alto Nanay, la cual se encontr¢ significativamente relacionada con ocupaciones realizadas fuera del centro poblado, residencia en localidades del r¡o Pintuyacu y lesiones cut neas.

Objectives. To determine the frequency, clinical features, and factors associated with M. ozzardi infection in volunteers of a malaria screening campaign in the district of Alto Nanay in the Amazon jungle of Peru. Materials and methods. A descriptive, analytical and cross-sectional study was performed. The participants were interviewed and examined by a physician and the data were recorded in a medical record. The diagnosis of M. ozzardi infection was performed using the method of thick blood smear and film. Results.The frequency of mansonelosis in this population was 47.8% (95% CI: 39.1 to 56.6). Through bivariate analysis we found that the report of decreased visual acuity or blurred vision and presence of subcutaneous tumors were the signs and symptoms statistically associated with the infection of microfilariae (p<0.05). Logistic regression found statistical association for residency in localities of the Pintuyacu or Alto Nanay rivers, employment in places far from the town center, the presence of subcutaneous tumors and skin thickening (p<0.05). Conclusions. There was a high number of mansonelosis by M. ozzardi in the district of Alto Nanay which was significantly related to working outside the town center, residing in the area of the Pintuyacu River, and skin lesions.

Mansonella ozzardi in the municipality of Tefé, Amazonas, Brazil, 60 years after the first report: an epidemiologic study

The prevalence of mansonelliasis was studied in the municipality of Tefé, state of Amazonas, Brazil. The prevalence (thick blood smear method) was 13.6% (147/1,078), higher in the Solimões River region (16.3%) than in the Tefé River region (6.3%). In the sampled communities in the Solimões River region, a higher density of cases was observed, as indicated by a kernel analysis (odds ratio 0.34; 95% confidence interval: 0.20-0.57). Males had a higher prevalence (χ2 = 31.292, p < 0.001) than women. Mansonella ozzardi prevalence was higher in retirees and farmers (28.9% and 27%, respectively). Prevalence also significantly increased with age (χ2 = -128.17, p < 0.001), with the highest numbers occurring in persons older than 67 years.

New records of Mansonella ozzardi: a parasite that is spreading from the state of Amazonas to previously uninfected areas of the state of Acre in the Purus River region

Mansonella ozzardi infections are common in the riverside communities along the Solimões, Negro and Purus Rivers in the state of Amazonas (AM). However, little is known about the presence of this parasite in communities located in regions bordering AM and the state of Acre. The prevalence rate of M. ozzardi infections was determined in blood samples from volunteers according to the Knott method. A total of 355 volunteers from six riverine communities were enrolled in the study and 65 (18.3%) were found to be infected with M. ozzardi. As expected, most of the infections (25%) occurred in individuals involved in agriculture, cattle rearing and fishing and an age/sex group analysis revealed that the prevalence increased beginning in the 40-50-years-of-age group and reached 33% in both sexes in individuals over 50 years of age. Based on the described symptomatology, articular pain and headache were found to be significantly higher among infected individuals (56 and 65% prevalence, respectively, p < 0.05). Sera from volunteers were subjected to ELISA using a cocktail of recombinant proteins from Onchocerca volvulus to evaluate the specificity of the test in an endemic M. ozzardi region. No cross-reactions between M. ozzardi-infected individuals and recombinant O. volvulus proteins were detected, thus providing information on the secure use of this particular cocktail in areas where these parasites are sympatric.

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. .

Mansonelliasis: a brazilian neglected disease

Mansonelliasis is a filariasis whose etiological agents are Mansonella ozzardi, Mansonella perstansand Mansonella streptocerca. Only the first two cited species occur in Brazil. M. ozzardi is widely distributed in Amazonas state and it is found along the rivers Solimões, Purus, Negro and their tributaries while M. perstans is restricted to the Upper Rio Negro. In this update, we report the occurrence of M. ozzardi in Amazonas since the 1950s, and we show that over the years this filariasis has been sustained with high prevalence, while maintaining a constant cycle of transmission in endemic areas due to the lack of treatment and control policies. M. perstans has so far only been recorded in indigenous populations in the Upper Rio Negro. However, the continuous flow of migrants to other regions may cause an expansion of this infection.

A mansonelose é uma filariose cujos agentes etiológicos são Mansonella ozzardi, M. perstans e M. strepotcerca. Somente as duas primeiras ocorrem no Brasil. M. ozzardi apresenta ampla distribuição no estado do Amazonas sendo encontrada ao longo dos rios Solimões, Purus e Negro e afluentes, ao passo que M. perstans possui distribuição restrita à região do Alto Rio Negro. Nesta atualização, é relatada a ocorrência de M. ozzardi no Amazonas desde a década de 1950 e, ao longo dos anos, esta filariose tem sustentado elevadas prevalências, mantendo um ciclo constante de transmissão nas áreas endêmicas em virtude da falta de políticas de tratamento e controle. Até o momento, M. perstans só foi registrada nas populações indígenas da região do Alto Rio Negro, mas, em razão da existência de um fluxo migratório contínuo para outras regiões, existe a possibilidade de sua expansão.

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.