RESUMO
BACKGROUND: Mansonellosis is an undermapped insect-transmitted disease caused by filarial nematodes that are estimated to infect hundreds of millions of people. Despite their prevalence, there are many outstanding questions regarding the general biology and health impacts of the responsible parasites. Historical reports suggest that the Colombian Amazon is endemic for mansonellosis and may serve as an ideal location to pursue these questions. METHODS: We deployed molecular and classical approaches to survey Mansonella prevalence among adults belonging to indigenous communities along the Amazon River and its tributaries near Leticia, Colombia. RESULTS: Loop-mediated isothermal amplification (LAMP) assays on whole-blood samples detected a much higher prevalence of Mansonella ozzardi infection (approximately 40%) compared to blood smear microscopy or LAMP performed using plasma, likely reflecting greater sensitivity and the ability to detect low microfilaremias and occult infections. Mansonella infection rates increased with age and were higher among men. Genomic analysis confirmed the presence of M. ozzardi that clusters closely with strains sequenced in neighboring countries. We successfully cryopreserved M. ozzardi microfilariae, advancing the prospects of rearing infective larvae in controlled settings. CONCLUSION: These data suggest an underestimation of true mansonellosis prevalence, and we expect that these methods will help facilitate the study of mansonellosis in endemic and laboratory settings.
Assuntos
Mansonelose , Parasitos , Masculino , Adulto , Animais , Humanos , Mansonella/genética , Mansonelose/epidemiologia , Mansonelose/parasitologia , Colômbia/epidemiologia , PrevalênciaRESUMO
OBJECTIVE: To assess the emergent zoonotic disease risk posed by the voracious human-biting blackfly species Simulium oyapockense in the peripheral regions of an expanding urban centre situated deep in the Brazilian Amazon rainforest. METHODS: We performed nine human landing catches at three periurban sites surrounding the Brazilian Amazon town of São Gabriel da Cachoeira. Using the detection of non-human primate filarial parasites as an indicator of the zoonotic disease threat posed by a biting insect, we screened 3328 S. oyapockense blackflies for the presence of zoonotic filarial DNA with an ITS-1 PCR assay and Sanger sequencing. RESULTS: Between 98 and 100% of the biting insects captured during our nine collections were identified as S. oyapockense; at our three collection sites and during our three seasonally-distinct collections this species was captured at rates between 28 and 294 blackflies per hour. PCR screening of the march-collected S. oyapockense detected infectious-stage (L3) Mansonella mariae parasites (which are only known to infect non-human primates) in >0.15% of the tested head samples. CONCLUSIONS: Our results show that residents of the periurban regions of São Gabriel da Cachoeira are routinely exposed to the bites of S. oyapockense blackflies which have previously fed on non-human primates.
Assuntos
Insetos Vetores/parasitologia , Mansonella/isolamento & purificação , Mansonelose/veterinária , Simuliidae/parasitologia , Zoonoses/transmissão , Animais , Mansonelose/parasitologia , Mansonelose/transmissão , Zoonoses/parasitologiaRESUMO
We reviewed Giemsa-stained thick blood smears, obtained through the national malaria surveillance program in the Amazon region of Ecuador, by light microscopy for Mansonella spp. microfilariae. Of 2,756 slides examined, 566 (20.5%) were positive. Nested PCR confirmed that the microfilariae were those of M. ozzardi nematodes, indicating that this parasite is endemic to this region.
Assuntos
Mansonella , Mansonelose/epidemiologia , Mansonelose/parasitologia , Animais , Equador/epidemiologia , Feminino , Geografia Médica , Humanos , Masculino , Mansonella/genética , Mansonella/isolamento & purificação , Mansonelose/diagnóstico , Reação em Cadeia da Polimerase , Prevalência , Vigilância em Saúde PúblicaRESUMO
BACKGROUND: In the Brazilian Amazon, the filarial nematode Mansonella ozzardi co-exists with malaria parasites and thick blood smear microscopy is considered the diagnostic gold standard. Transfusion of M. ozzardi microfilariae does not establish new infections, however microfilariae can survive approximately 2 years in blood-recipients with unknown risk of pathology. Data on transfusion-transmitted filariasis are lacking. This study investigated M. ozzardi parasitemias in blood donors from decentralized centers of "Fundação Hematologia e Hemoterapia do Estado do Amazonas/HEMOAM," Northern Brazil. STUDY DESIGN AND METHODS: Cross-sectional investigation employing blood smear microscopy (n = 356) and qualitative nested-M. ozzardi-PCR (227 out of 356) in donor candidates from 19 hemocenters in interior/rural municipalities of Amazonas state. FINDINGS: Participants were mostly young males. Positivity by microscopy was 7.9% (28 out of 356) and 23.8% by M. ozzardi-PCR (54 out of 227). Parasitaemias were found in 16 out of 19 municipalities. In 54 M. ozzardi-positives, 24 were ineligible; among 30 that donated, 27 were interdicted by seropositivity (22 anti-HBc, 3 anti-HBc + HBsAg, 1 Chagas+malaria, 1 VDRL). Seropositivty was higher in M. ozzardi-PCR-positives vs M. ozzardi-PCR-negatives (OR = 15.8, 95% CI 4.5-56.1, p < 0.0001). Three M. ozzardi contaminated blood units were transfused, but no follow-up information on the recipients is available. MAIN CONCLUSIONS: Our study provides important baseline data on M. ozzardi among blood donors from the Brazilian Amazon. Further investigations in endemic areas are necessary to clarify possible association between M. ozzardi and other infections and also to elucidate whether there is any significant clinical effect upon transfusion of contaminated blood.
Assuntos
Doadores de Sangue/estatística & dados numéricos , Mansonella/patogenicidade , Mansonelose/parasitologia , Adolescente , Adulto , Idoso , Animais , Brasil/epidemiologia , Estudos Transversais , Feminino , Humanos , Masculino , Mansonella/isolamento & purificação , Mansonelose/epidemiologia , Pessoa de Meia-Idade , Adulto JovemRESUMO
Despite the broad distribution of M. ozzardi in Latin America and the Caribbean, there is still very little DNA sequence data available to study this neglected parasite's epidemiology. Mitochondrial DNA (mtDNA) sequences, especially the cytochrome oxidase (CO1) gene's barcoding region, have been targeted successfully for filarial diagnostics and for epidemiological, ecological and evolutionary studies. MtDNA-based studies can, however, be compromised by unrecognised mitochondrial pseudogenes, such as Numts. Here, we have used shot-gun Illumina-HiSeq sequencing to recover the first complete Mansonella genus mitogenome and to identify several mitochondrial-origin pseudogenes. Mitogenome phylogenetic analysis placed M. ozzardi in the Onchocercidae "ONC5" clade and suggested that Mansonella parasites are more closely related to Wuchereria and Brugia genera parasites than they are to Loa genus parasites. DNA sequence alignments, BLAST searches and conceptual translations have been used to compliment phylogenetic analysis showing that M. ozzardi from the Amazon and Caribbean regions are near-identical and that previously reported Peruvian M. ozzardi CO1 reference sequences are probably of pseudogene origin. In addition to adding a much-needed resource to the Mansonella genus's molecular tool-kit and providing evidence that some M. ozzardi CO1 sequence deposits are pseudogenes, our results suggest that all Neotropical M. ozzardi parasites are closely related.
Assuntos
Complexo IV da Cadeia de Transporte de Elétrons/genética , Genoma Mitocondrial , Mansonella/classificação , Mansonella/genética , Mansonelose/parasitologia , Pseudogenes , Animais , Genômica/métodos , Humanos , Filogenia , RNA Ribossômico/genética , RNA Ribossômico 5S/genéticaRESUMO
The treatment of mansonelliasis is still a challenge because there are few clinical trials for the treatment of the disease. This double-blind, randomized, placebo-controlled study (phase III clinical trial) was conducted to evaluate the effectiveness of a single oral dose of ivermectin (0.15 mg/kg) in the reduction of the Mansonella ozzardi microfilaraemia and the occurrence of adverse effects in infected people compared with the control group treated with placebo. A total of 49 microfilaraemic patients were randomly selected from the municipality of Lábrea, State of Amazonas, in the Brazilian Amazon. Among them, 40 patients have concluded the study, 19 treated with ivermectin and 21 treated with placebo. In the first and third days after the treatment, all the patients were clinically evaluated, and the diagnostic and quantification of blood microfilariae through blood filtration in polycarbonate membranes was performed. A significant reduction of the microfilaraemia (99.9%) was observed in the patients who received ivermectin. Slight changes in laboratory test results, without clinical importance, were seen in treated and control groups. Our results suggest that ivermectin is effective and safe for the treatment of infections caused by M. ozzardi.
Assuntos
Anti-Helmínticos/uso terapêutico , Ivermectina/uso terapêutico , Mansonella/efeitos dos fármacos , Mansonelose/tratamento farmacológico , Microfilárias/efeitos dos fármacos , Administração Oral , Adolescente , Adulto , Animais , Brasil , Método Duplo-Cego , Esquema de Medicação , Feminino , Humanos , Masculino , Mansonella/crescimento & desenvolvimento , Mansonelose/parasitologia , Microfilárias/crescimento & desenvolvimento , Pessoa de Meia-Idade , Resultado do TratamentoRESUMO
Mansonella ozzardi (Nematoda: Onchocercidae) is an understudied filarial nematode, originally described by Patrick Manson in 1897, that can be transmitted by two families of dipteran vectors, biting midges (most of them members of the genus Culicoides) and black flies (genus Simulium). With a patchy geographic distribution from southern Mexico to northwestern Argentina, human infection with M. ozzardi is highly prevalent in some of the Caribbean islands, along riverine communities in the Amazon Basin, and on both sides of the border between Bolivia and Argentina. There is no clinical entity unequivocally associated with M. ozzardi infection, although fever, arthralgia, headache, cold lower extremities, and itchy cutaneous rashes are occasionally mentioned in case report series. More recently, ocular manifestations (especially keratitis) have been associated with mansonelliasis, opening an important area of investigation. Here, we briefly review the biology, epidemiology, pathogenesis, and clinical aspects of M. ozzardi infection and point to some existing knowledge gaps, aiming to stimulate a research agenda to help filling them.
Assuntos
Mansonella , Mansonelose/epidemiologia , Mansonelose/parasitologia , Doenças Negligenciadas/epidemiologia , Doenças Negligenciadas/parasitologia , Animais , Anti-Helmínticos/farmacologia , Anti-Helmínticos/uso terapêutico , Vetores Artrópodes/classificação , Vetores Artrópodes/parasitologia , Humanos , Mansonella/efeitos dos fármacos , Mansonella/crescimento & desenvolvimento , Mansonella/isolamento & purificação , Mansonella/fisiologia , Mansonelose/diagnóstico , Mansonelose/terapia , Doenças Negligenciadas/diagnóstico , Doenças Negligenciadas/terapia , PrevalênciaRESUMO
Mansonellosis is endemic in several regions of Africa, the Caribbean, and Latin America. Mansonella ozzardi and Mansonella perstans have been reported in Latin America, including the Amazon region. A morphological and molecular microfilariae study was performed in Pauini (Brazil). Blood samples were collected from 40 individuals, and were analyzed by Giemsa-stained blood film and by two different nested polymerase chain reactions which detect internal transcribed spacer-1 and the major sperm protein gene. By microscopy, 14 of 40 were positive: 11 as M. ozzardi and three as M. perstans-like infections. Both molecular methods detected 19 positive cases as M. ozzardi, including those 14 individuals detected by microscopy, without detectable genetic differences among any of the 19 positive samples. Molecular techniques showed an improvement of mansonellosis diagnosis and may become an effective tool to evaluate the present status of M. ozzardi and M. perstans in Latin America.
Assuntos
Mansonella , Mansonelose/parasitologia , Microfilárias , Animais , Brasil/epidemiologia , Humanos , Mansonella/genética , Mansonella/ultraestrutura , Mansonelose/diagnóstico , Mansonelose/epidemiologia , Microfilárias/genética , Microfilárias/ultraestrutura , Microscopia , Filogenia , Reação em Cadeia da PolimeraseRESUMO
Human filariasis caused by Mansonella ozzardi is a parasitic infection of a controversial pathology and poorly studied in Argentina. The aim of this study was not only to obtain data of the prevalence of M. ozzardi in Northwestern Argentina through comparison of infection rates in relation to sex and age group determining the range of distribution of mansonelliasis in the region but also to investigate the prevalence of the cases over time. Through field work carried between 1986 and 2010 by technicians of the National Ministry of Health, aiming to detect active cases of malaria in the Northwestern Argentina, blood samples were taken for smear and thick blood. 417 blood samples were examined, 381 of them (91.4%) were positive for M. ozzardi. The highest prevalence was found in Salta province (92.3%) mainly affecting the male sex (92.6%) and the age groups comprised between 48-57 years (97.0%) and ≥ 68 (90.7%). The paired t-test and Spearman coefficients showed significant differences in prevalence according to sex (t=2.677; p=0.015), and according infection rates in males and females/age (r=0.994; p=0.001; r=0.994, p=0.001, respectively). The prevalence over time showed a general pattern with the highest cases in 1986, decreasing later during the following years. Aguas Blancas, El Oculto and San Ramón de la Nueva Orán, exhibited a pattern of prevalence according to the general trend but Salvador Mazza and Tartagal, which are also localities close to the border with Bolivia, showed peaks of cases up to 2010. These results provide relevant information about M. ozzardi in Argentina, demonstrating not only its presence and endemicity after almost 100 years from its discovery in the country, but also its wide range of distribution in the region.
Assuntos
Sangue/parasitologia , Mansonella/isolamento & purificação , Mansonelose/epidemiologia , Mansonelose/parasitologia , Adolescente , Adulto , Fatores Etários , Idoso , Idoso de 80 Anos ou mais , Animais , Argentina/epidemiologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Prevalência , Floresta Úmida , Fatores Sexuais , Fatores de Tempo , Clima Tropical , Adulto JovemRESUMO
BACKGROUND: Mansonella ozzardi is a poorly understood human filarial parasite with a broad distribution throughout Latin America. Most of what is known about its parasitism has come from epidemiological studies that have estimated parasite incidence using light microscopy. Light microscopy can, however, miss lighter, submicroscopic, infections. In this study we have compared M. ozzardi incidence estimates made using light microscopy, with estimates made using PCR. METHODS: 214 DNA extracts made from Large Volume Venous Blood Samples (LVVBS) were taken from volunteers from two study sites in the Rio Solimões region: Codajás [n = 109] and Tefé [n = 105] and were subsequently assayed for M. ozzardi parasitism using a diagnostic PCR (Mo-dPCR). Peripheral finger-prick blood samples were taken from the same individuals and used for microscopic examination. Finger-prick blood, taken from individuals from Tefé, was also used for the creation of FTAcard dried blood spots (DBS) that were subsequently subjected to Mo-dPCR. RESULTS: Overall M. ozzardi incidence estimates made with LVVBS PCRs were 1.8 times higher than those made using microscopy (44.9% [96/214] compared with 24.3% [52/214]) and 1.5 times higher than the PCR estimates made from FTAcard DBS (48/105 versus 31/105). PCR-based detection of FTAcard DBS proved 1.3 times more sensitive at diagnosing infections from peripheral blood samples than light microscopy did: detecting 24/105 compared with 31/105. PCR of LVVBS reported the fewest number of false negatives, detecting: 44 of 52 (84.6%) individuals diagnosed by microscopy; 27 of 31 (87.1%) of those diagnosed positive from DBSs and 17 out of 18 (94.4%) of those diagnosed as positive by both alternative methodologies. CONCLUSIONS: In this study, Mo-dPCR of LVVBS was by far the most sensitive method of detecting M. ozzardi infections and detected submicroscopic infections. Mo-dPCR FTAcard DBS also provided a more sensitive test for M. ozzardi diagnosis than light microscopy based diagnosis did and thus in settings where only finger-prick assays can be carried-out, it may be a more reliable method of detection. Most existing M. ozzardi incidence estimates, which are often based on light microscope diagnosis, are likely to dramatically underestimate true M. ozzardi parasitism incidence levels.