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Mansonella ozzardi, a filarioid parasite, causes human mansonellosis in the Americas. We identified raccoons (Procyon lotor) as wildlife reservoirs of M. ozzardi in Costa Rica. Noting the sympatry of free-ranging raccoons and humans, we conclude that mansonellosis is a considerable public health risk in the region.

Mansonella spp. have been reported to have a wide global distribution. Despite the distribution and co-occurrence with other filarial parasites like Wuchereria bancrofti, Onchocerca volvulus and Loa loa, it is given little attention. There are few surveillance programmes for assessing the distribution of mansonellosis, due to the associated mild to no symptoms experienced by infected people. However, addressing this infection is critical to the onchocerciasis control program as current rapid diagnostic tools targeting O. volvulus have the tendency to cross react with Mansonella species. In this study we identified and characterised M. perstans from five sites in two districts in the Volta Region of Ghana and compared them to samples from other regions. Night blood smears and filter blood blots were obtained from individuals as part of a study on lymphatic filariasis. The Giemsa-stained smears were screened by microscopy for the presence of filarial parasites. Genomic DNA was extracted from blood blots from 39 individuals that were positive for M. perstans and Nested PCR targeting the internal spacer 1 (ITS-1) was conducted. Of these, 30 were sequenced and 24 sequences were kept for further analysis. Phylogenetic analysis of 194 nucleotide positions showed no differences in the samples collected. The similarities suggests that there could be one species in this area. However, more robust studies with larger sample sizes are required to draw such conclusions. We also observed a clustering of the samples from Ghana with reference sequences from Africa and Brazil, suggesting they could be related. This study draws further attention to a neglected infection, presents the first characterisation of M. perstans in Ghana and calls for more population-based studies across different geographical zones to ascertain species variations and disease distribution., Competing Interests: The authors have declared that no competing interest exist., (Copyright: © 2024 Opoku, de Souza. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Background: Life-style metabolic diseases are steadily rising, not only in developed countries, but also in low- and middle-income countries, presenting a global health problem. Metabolic disorders like type 2 diabetes and cardiovascular diseases are among the ten leading causes of death defined by the WHO in 2019. Results from animal and observational human studies suggest a connection between the decline in human helminth infections and rise of life-style-associated metabolic diseases in developing regions. This trial was designed to investigate filarial infections and their impact on metabolic diseases in Cameroon. We hypothesize that the induction of regulatory immune responses during filarial infection reduces obesity-induced low-grade inflammatory immune responses and thereby improves metabolic parameters, whereas anthelmintic treatment abolishes this protective effect., Methods/design: Participants infected with Mansonella perstans, Onchocerca volvulus and/or Loa loa being lean (BMI <25), overweight (BMI >25 and <30) or clinically obese (BMI ≥30) from Littoral regions of Cameroon will be evaluated for their parasitological, immunological, metabolic and biochemical profile before and after treatment of their parasitic infections. Anthropomorphic measurements and a detailed questionnaire will complement our analysis. The investigation will assess blood immune cell populations, serum adipokines and cytokines that could be influenced by the parasite infection and/or metabolic diseases. Further, parameters like blood glucose, homeostatic model assessment of insulin resistance (HOMA-IR), circulating lipids and circulating makers of liver function will be monitored. Parameters will be assessed before treatment, 12 and 18 months after treatment., Conclusion: The focus of this study is to obtain a comprehensive metabolic profile of the participants in rural areas of Cameroon and to investigate the relationship between filarial immunomodulation and metabolic diseases. This study will elucidate the effect of anti-filarial treatment on the metabolic and immunological parameters that partake in the development of insulin resistance, narrowing in on a potential protective effect of filarial infections on metabolic diseases., Trial Registration: doi.org/10.1186/ISRCTN43845142, ISRCTN43845142 February 2020 Trial title Effects of filarial parasite infection on type 2 diabetes Issue date: 27.10.22, V.1., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Lenz et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Objectives: Loa loa and Mansonella perstans are two very common filarial species in Africa. Although microscopy is the traditional diagnostic method for human filariasis, several polymerase chain reaction (PCR) methods have emerged as an alternative approach for identifying filarial parasites. The aim of this study is to compare three molecular methods and decide which is the most suitable for diagnosing human loiasis and mansonellosis in non-endemic regions using dried blood spot (DBS) as a medium for sample collection and storage., Methods: A total of 100 DBS samples, with their corresponding thin and thick blood smears, were selected for this study. Microscopy was used as the reference method to diagnose and calculate the microfilaraemia. Filarial DNA was extracted using the saponin/Chelex method and the DNA isolated was assayed by Filaria-real time-PCR, filaria-nested PCR, and cytochrome oxidase I PCR. All PCR products were subsequently purified and sequenced. The statistical values for each molecular test were calculated and compared., Results: Overall, 64 samples were identified as negative by all tests and a further 36 samples were positive by at least one of the methods used. The sensitivity and specificity were similar for the different molecular methods, all of which demonstrated good agreement with microscopy., Conclusions: Based on this study, and from a practical point of view (single and short amplification round), the optimal technique for diagnosing filarial infection in non-endemic regions is filaria-real time-PCR, which presents high sensitivity and specificity and is also able to detect a wide range of human filariae., (© 2022 John Wiley & Sons Ltd.)

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., (Copyright © 2022 The Authors. Published by Elsevier B.V. All rights reserved.)

Background: Loa loa and Mansonella perstans-the causative agents of loiasis and mansonellosis-are vector-borne filarial parasites co-endemic in sub-Saharan Africa. Diagnosis of both infections is usually established by microscopic analysis of blood samples. It was recently established that the odds for detecting Plasmodium spp. is higher in capillary (CAP) blood than in venous (VEN) blood. In analogy to this finding this analysis evaluates potential differences in microfilaraemia of L. loa and M. perstans in samples of CAP and VEN blood., Methods: Recruitment took place between 2015 and 2019 at the CERMEL in Lambaréné, Gabon and its surrounding villages. Persons of all ages presenting to diagnostic services of the research center around noon were invited to participate in the study. A thick smear of each 10 microliters of CAP and VEN blood was prepared and analysed by a minimum of two independent microscopists. Differences of log2-transformed CAP and VEN microfilaraemia were computed and expressed as percentages. Furthermore, odds ratios for paired data were computed to quantify the odds to detect microfilariae in CAP blood versus in VEN blood., Results: A total of 713 participants were recruited among whom 52% were below 30 years of age, 27% between 30-59 years of age and 21% above 60 years of age. Male-female ratio was 0.84. Among 152 participants with microscopically-confirmed L. loa infection median (IQR) microfilaraemia was 3,650 (275-11,100) per milliliter blood in CAP blood and 2,775 (200-8,875) in VEN blood (p<0.0001), while among 102 participants with M. perstans this was 100 (0-200) and 100 (0-200), respectively (p = 0.44). Differences in linear models amount up to an average of +34.5% (95% CI: +11.0 to +63.0) higher L. loa microfilaria quantity in CAP blood versus VEN blood and for M. perstans it was on average higher by +24.8% (95% CI: +0.0 to +60.5). Concordantly, the odds for detection of microfilaraemia in CAP samples versus VEN samples was 1.24 (95% CI: 0.65-2.34) and 1.65 (95% CI: 1.0-2.68) for infections with L. loa and M. perstans, respectively., Conclusion: This analysis indicates that average levels of microfilaraemia of L. loa are higher in CAP blood samples than in VEN blood samples. This might have implications for treatment algorithms of onchocerciasis and loiasis, in which exact quantification of L. loa microfilaraemia is of importance. Furthermore, the odds for detection of M. perstans microfilariae was higher in CAP than in VEN blood which may pre-dispose CAP blood for detection of M. perstans infection in large epidemiological studies when sampling of large blood quantities is not feasible. No solid evidence for a higher odds of L. loa microfilariae detection in CAP blood was revealed, which might be explained by generally high levels of L. loa microfilaraemia in CAP and VEN blood above the limit of detection of 100 microfilariae/ml. Yet, it cannot be excluded that the study was underpowered to detect a moderate difference., Competing Interests: The authors have declared that no competing interests exist.

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.

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.

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 .

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.

We report 74 patients in Italy infected with Mansonella perstans nematodes, a poorly described filarial parasite. M. perstans nematodes should be included in the differential diagnosis for patients with eosinophilia from disease-endemic countries. Serologic analysis is useful for screening, and testing for microfilaremia in peripheral blood should be performed for parasite-positive patients.

We obtained ribosomal and mitochondrial DNA sequences from residents of Amazonas state, Brazil, with Mansonella parasitemias. Phylogenetic analysis of these sequences confirm that M. ozzardi and M. perstans parasites occur in sympatry and reveal the close relationship between M. perstans in Africa and Brazil, providing insights into the parasite's New World origins.

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., (© The American Society of Tropical Medicine and Hygiene.)

The study of the interactions among parasites within their hosts is crucial to the understanding of epidemiology of disease and for the design of effective control strategies. We have conducted an assessment of infections with Loa loa, Mansonella perstans, Wuchereria bancrofti, and Plasmodium falciparum in eastern Cameroon using a highly sensitive and specific quantitative polymerase chain reaction assay using archived dried whole blood spots. The resident population (N = 1,085) was parasitized with M. perstans (76%), L. loa (39%), and P. falciparum (33%), but not with W. bancrofti Compared with single infections (40.1%), coinfection was more common (48.8%): 21.0% had L. loa-M. perstans (Ll(+)/Mp(+)/Pf(-)), 2.7% had L. loa-P. falciparum (Ll(+)/Pf(+)/Mp(-)), 15.1% had M. perstans-P. falciparum (Mp(+)/Pf(+)/Ll(-)), and 10.0% had L. loa-M. perstans-P. falciparum (Ll(+)/Mp(+)/Pf(+)). Interestingly, those with all three infections (Ll(+)/Mp(+)/Pf(+)) had significantly higher L. loa microfilaria (mf) counts than either single Ll(+) (P = 0.004) or double Ll(+)/Mp(+) (P = 0.024) infected individuals. Of those infected with L. loa, the mean estimated counts of L. loa mf varied based on location and were positively correlated with estimated intensities of M. perstans mf. Finally, at a community level, heavy L. loa infections were concentrated in a few individuals whereby they were likely the major reservoir for infection., (© The American Society of Tropical Medicine and Hygiene.)

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.

Background: Mansonella perstans is a widespread, but relatively unknown human filarial parasite transmitted by Culicoides biting midges. Although it is found in many parts of sub-Saharan Africa, only few studies have been carried out to deepen the understanding of its ecology, epidemiology, and health consequences. Hence, knowledge about ecological drivers of the vector and parasite distribution, integral to develop spatially explicit models for disease prevention, control, and elimination strategies, is limited., Methodology: We analyzed data from a comprehensive nationwide survey of M. perstans infection conducted in 76 schools across Uganda in 2000-2003, to identify environmental drivers. A suite of Bayesian geostatistical regression models was fitted, and the best fitting model based on the deviance information criterion was utilized to predict M. perstans infection risk for all of Uganda. Additionally, we investigated co-infection rates and co-distribution with Wuchereria bancrofti and Plasmodium spp. infections observed at the same survey by mapping geographically overlapping areas., Principal Findings: Several bioclimatic factors were significantly associated with M. perstans infection levels. A spatial Bayesian regression model showed the best fit, with diurnal temperature range, normalized difference vegetation index, and cattle densities identified as significant covariates. This model was employed to predict M. perstans infection risk at non-sampled locations. The level of co-infection with W. bancrofti was low (0.3%), due to limited geographic overlap. However, where the two infections did overlap geographically, a positive association was found., Conclusions/significance: This study presents the first geostatistical risk map for M. perstans in Uganda. We confirmed a widespread distribution of M. perstans, and identified important potential drivers of risk. The results provide new insight about the ecologic preferences of this otherwise poorly known filarial parasite and its Culicoides vector species in Uganda, which might be relevant for other settings in sub-Saharan Africa.

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.

The occurrence of Mansonella perstans was studied among people who had resided in Calabar metropolis for at least one year prior to the time of this study, which lasted from February to August, 2011. One thousand residents, comprising 530 males and 470 females, with an age range of 4 to 59 years, participated in the study, after an informed consent. Two millilitres of venous blood were collected at day time into EDTA bottles. The blood samples were processed by diluting 1ml of blood in 9 mls of 1% formalin and centrifuging the preparation at 3,000 revolutions per minute for 5 minutes. Deposits were left as blood smears on clean slides and after air-drying, were stained with 3% Giemsa solution for microscopy. Overall, the prevalence of Mansonella perstans was 2% (20 vs 1,000). The prevalence of the parasite among the females was 2.1% (10 vs 470) and males 1.9% (10 vs 530). Microfilaraemia only occurred among subjects aged between 25 and 45 years. The results of this study suggest that loiasis, which earned Calabar an unenviable mention in early medical literature, is no longer very prevalent in the area. Instead, a low prevalence of mansonelliasis occurs, in tandem with the existence of sparse farmlands of banana and plantain. Old stems of these crops provide suitable habitats for Culicoides species, which transmit Mansonella perstans.

An epidemiologic (cross-sectional study) survey on 462 inhabitants in Corail, Haiti showed that 16.5% were infected with Mansonella ozzardi. This finding was determined from a single 20-μL sample of finger prick blood from each person. Among children, < 2% had a detectable microfilaremia. In persons > 15 years of age, the prevalence of infection for males and females was 23% and 21%, respectively. In general, the microfilaremias were low and 70% of positive persons had < 10 microfilariae per 20 μL of blood; only 5% had > 50 microfilariae. This study shows that persons living near mangrove marshes that are breeding sites for Culicoides furens and C. barbosai biting midges, which are recognized vectors of M. ozzardi in Haiti, are consequently more frequently infected than those living in downtown area of Corail or inland., (© The American Society of Tropical Medicine and Hygiene.)

During August 2010-December 2012, we conducted a study of patients in Ghana who had Buruli ulcer, caused by Mycobacterium ulcerans, and found that 23% were co-infected with Mansonella perstans nematodes; 13% of controls also had M. perstans infection. M. perstans co-infection should be considered in the diagnosis and treatment of Buruli ulcer.

A case of co-infection with Plasmodium vivax and Mansonella ozzardi was detected in a blood sample from a person who had shown symptoms of malaria and lived in a city that was close to the Argentina/Bolivia border. The case was detected during a random revision of thick and thin smears from patients diagnosed with malaria from various towns and cities located in north-western Argentina between 1983 and 2001. Trophozoites of P. vivax were observed in the thin blood smear along with M. ozzardi microfilaria (larval form), which presented a long, slender, pointed anucleate tail and the absence of the sheath. This last characteristic is shared with Mansonella perstans, Mansonella streptocerca and Onchocerca volvulus. More rigorously controlled studies to detect other co-infection cases in the area as well as the possibility of importation from Bolivia into Argentina are currently ongoing. The relationship between the malaria parasite and microfilaria, the potential effect of malaria treatment on the development of M. ozzardi, and the possible impact of this microfilaria on the immunity of a person against P. vivax are all still unknown. This contribution constitutes a point of focus for future studies involving the interaction between the parasites and the potential risk that humans are exposed to.

DNA sequence comparisons are useful for characterizing proposed new parasite species or strains. Microfilariae with an atypical arrangement of nuclei behind the cephalic space have been recently described in human blood samples from the Amazon region of Peru. Three blood specimens containing atypical microfilariae were genetically characterized using three DNA markers (5S ribosomal DNA, 12S ribosomal DNA, and cytochrome oxidase I). All atypical microfilariae were clustered into the Mansonella group and indistinguishable from M. ozzardi based on these DNA markers.

Background & Objectives: The study was aimed at elucidating the prevalence and intensity of Mansonella perstans microfilaraemia in the Emohua Local Government Area, Nigeria, and ascertaining the abundance, circadian, and the annual biting patterns of the Culicoides vector., Methods: Thick smear of 50 µl finger-prick blood stained with Giemsa was examined microscopically in a cross-sectional study. Vector landing collection on human bait was employed in a longitudinal study of the vector biting patterns, carried out between July 2005 and August 2006., Results: Of 1486 individuals examined, 11.2% of both males and females were positive for M. perstans microfilaraemia. Microfilaraemia appeared early in life. The overall geometric mean intensity among those with positive microfilaraemia was 117 mf/ml (121 mf/ml for males and 113 mf/ml for females). The differences in geometric mean intensity between different age groups were statistically significant (one-way analysis of variance; p <0.05), being highest in the oldest age group (266 mf/ml). A total of 1183 female Culicoides sp were caught from September 2005 to August 2006. The abundance of Culicoides sp was seasonal. The circadian biting activity had a broad peak between 0700 and 1200 hrs. The monthly biting rates ranged from zero bite per person per month in January 2006 to 1151 bites per person per month in June 2006. The annual biting rate was 7382 bites per person per year., Conclusion: Majority of those with positive microfilaraemia were poor socioeconomically, underscoring the need for health education and application of effective control measures against Culicoides biting midges in Emohua.

The prevalence and intensity of Loa loa and Mansonella perstans were studied in three villages of the Ijebu north area in Ogun State, western Nigeria. Blood samples were collected by finger-pricking from 373 (181 males, 192 females) subjects with an age range of 4 to 55 years. The blood samples were examined microscopically for the presence of microfilariae (mf). L. loa and M. perstans were present in the blood samples in 39 (10.5%) and 12 (3.2%) of the subjects, respectively. Neither of the infections were found to be sex-dependent. The geometric mean of the L. loa intensity was 1.8 mf per 50 microl of blood, while that of M. perstans was 1.5 mf per 50 microl. The prevalence of mixed infections of both L. loa and M. perstans was 1.0%.

The long-term effect of a single oral dose of 150 microg/kg of body weight of ivermectin on Mansonella streptocerca microfilariae was studied in western Uganda. Before treatment, the geometric mean microfilaria density (mf) in 93 infected persons was 2.4 mf/mg of skin (range, 0.1-42.6). One year after treatment, 43 persons (46%) were microfilaria-negative, and the geometric mean in the remaining persons dropped significantly, to 0.7 mf/mg (range, 0.1-6.9). Thus, ivermectin is highly effective against M. streptocerca, and a single dose leads to a sustained suppression of microfilariae in skin. In Africa, ivermectin is used for mass treatment to control Onchocerca volvulus and Wuchereria bancrofti. Because these filarial parasites are often coendemic with M. streptocerca, the treated population may receive the additional benefit of suppression of M. streptocerca microfilariae.

To differentiate the skin-dwelling filariae Mansonella streptocerca and Onchocerca volvulus, a nested polymerase chain reaction (PCR) assay was developed from small amounts of parasite material present in skin biopsies. One nonspecific and one specific pair of primers were used to amplify the 5S rDNA spacer region of M. streptocerca. Biopsies with different microfilaria densities obtained from 104 Ugandans living in an area endemic for M. streptocerca were tested using both the nested PCR assay and standard parasitologic assessment of microfilariae. All 82 samples from microfilaria carriers were positive when tested using the nested PCR assay. In addition, M. streptocerca DNA could be detected in 16 samples thought to be microfilaria negative. Furthermore, six days following ivermectin treatment, M. streptocerca DNA was found in 12 of 14 microfilaria-negative biopsies. Control skin samples from patients infected with O. volvulus were all negative in the nested PCR assay. This assay improves the diagnosis of M. streptocerca and will facilitate further epidemiologic studies.

We studied the short-term effects of a single dose of 150 micrograms/kg body weight ivermectin on Mansonella streptocerca in an area endemic for streptocerciasis, but not for onchocerciasis, in western Uganda. Six and 12 days after treatment no microfilaria (mf) were found in the skin of 53 out of 96 mf carriers living in 3 villages, and the geometric means of the mf densities of remaining mf carriers were only 33-40% of pretreatment levels. This reduction of mf density was highly significant (P < 0.0001). Immunohistological examination of skin biopsies showed degenerated and disintegrating mf surrounded by activated eosinophils (positive for activated cationic protein), macrophages, and neutrophils (positive for myeloperoxidase and defensin) on day 6 after treatment. Remarkable was the invasion of young, L1 protein-positive macrophages and the release of neutrophil defensin as signs of acute inflammation. We conclude that ivermectin has a strong microfilaricidal activity against M. streptocerca. Common adverse effects were increased pruritus and acute papular dermatitis in 45% of 86 mf carriers on day 6 after treatment. No serious adverse side-effects were noticed in about 700 treated persons.

The prevalence of filariae in wild raccoons trapped in southeast Georgia was determined. Examination of blood samples revealed that 74 of 113 raccoons (66%) trapped in 6 southeastern Georgia counties were infected. Seventy-three of these raccoons (65%) were infected with Mansonella llewellyni and this parasite was observed in raccoons from every location examined. Dirofilaria tenuis was found in 22 raccoons (20%) and was observed in only 3 of the 6 counties surveyed. An adult specimen of Acanthocheilonema procyonis was found in the subcutaneous tissues of 1 of 5 necropsied raccoons. This is the first record of filariae in raccoons from Georgia. In addition, Dirofilaria-like larvae were found in Aedes taeniorhyncus mosquitoes collected in Liberty County.

Parasitic infection as the only or concomitant cause of infertility in Caucasian women is rare. A parasitic infection may also present itself quite unexpectedly as a coincidental finding as shown with this case report. Moving microfilariae of Mansonella perstans were found in the aspirated follicular fluid of a patient who underwent in-vitro fertilization (IVF) with embryo transfer because of tubal pathology due to Chlamydia trachomatis. The patient also appeared to have a Schistosoma infection. To our knowledge, the presence of parasites in follicular fluid has never been reported before. We expect that infertility physicians may be confronted with parasitic infections more often, not only in patients originating from tropical countries but also in Western women as a result of a tendency to travel more frequently to exotic and (sub)tropical countries.

This report summarizes the findings of the 17 published studies involving humans who have been experimentally infected with filarial parasites. Over the past 60 years, 45 individuals have been deliberately infected with Wuchereria bancrofti, Brugia malayi, Brugia pahangi, Loa loa, Mansonella perstans, Mansonella ozzardi, and/or Onchocerca volvulus. The findings from these experimental infections of humans have helped define microfilarial survival and periodicity within human hosts, the prepatent period for the causative agents of lymphatic filariasis, etiologic agents for particular clinical syndromes, immunologic and hematologic consequences of filarial infection, and the role of chemotherapeutic agents in the prevention and treatment of filarial infections.