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Acta Tropica Nov 2022Culicoides biting midges are capable to transmit Oropouche virus, Bluetongue virus and Mansonella spp. This study aimed to assess the utility of DNA barcode as an...
Culicoides biting midges are capable to transmit Oropouche virus, Bluetongue virus and Mansonella spp. This study aimed to assess the utility of DNA barcode as an alternative method in the Culicoides species identification. The study was conducted in Jamari National Forest. Biting midges were collected using HP light traps during four months, February, April, August and October 2018. Insects were morphologically identified to the species level, and rest of the body were subjected to DNA extraction and PCR targeting a fragment of the cytochrome c oxidase subunit I (COI) gene, which were analyzed and deposited in GenBank. A phylogenetic gene tree was reconstructed using RAxML software, and the sequences were assigned at Molecular Operational Taxonomic Unit (MOTU) level by species delimitation algorithms. According to morphological approach, 18 species of 2 subgenera and 7 species groups were identified. A total of 191 new COI barcodes from 18 species were generated. Of these, fifteen species have been deposited for the first time in all datasets in the world. These sequences allowed the correct identification of 188 and 187 specimens according to the BM and BCM criteria, respectively. The intraspecific genetic distances ranged from 0 to 16.5%, while the interspecific ones ranged from 2.1 to 27.1%. The nominal species Culicoides glabellus and C. tetrathyris splitted into three and two MOTUs, respectively, except for mPTP, indicating a cryptic diversity in these species. Also, sequences of C. pseudodiabolicus formed two MOTUs using all algorithms, except for PTP and ABGD, suggesting the existence of two potential species. In contrast, some barcodes of C. quasiparaensis and C. paraensis merged into a single MOTU, which can be explained by the complex characteristics of the paraensis group, since these species have similar morphological characters. Here, we provided the first COI barcodes for biting midges in Rondônia and Brazil, and demonstrates that these are sufficient to discriminate between some species.
Topics: Animals; Brazil; Ceratopogonidae; DNA; DNA Barcoding, Taxonomic; Phylogeny
PubMed: 35905777
DOI: 10.1016/j.actatropica.2022.106619 -
The American Journal of Tropical... Aug 2022Filarial infections caused by Loa loa and Mansonella perstans are a considerable public health burden in rural regions of Central Africa. Rapid diagnostic tools for the...
Filarial infections caused by Loa loa and Mansonella perstans are a considerable public health burden in rural regions of Central Africa. Rapid diagnostic tools for the detection of microfilariae in the blood are needed. Field's stain is a rapid staining technique for microscopic slides originally established for malaria diagnostics. It requires less than 1 minute of staining compared with conventional staining protocols requiring at least 15 to 20 minutes for staining and could thus significantly accelerate diagnostics for human filariasis. Here we evaluated Field's stain as a rapid staining technique in comparison to Giemsa stain for the detection of microfilariae in peripheral blood. Blood smears were collected from 175 participants residing in the region of Lambaréné and Fougamou, Gabon. Each participant's samples were stained in parallel with Field's stain and conventional Giemsa stain. Slides were then microscopically assessed and compared for qualitative and quantitative results by a blinded assessor for the two endemic filarial blood pathogens M. perstans and L. loa. Field's stain shows excellent diagnostic performance characteristics for L. loa microfilariae compared with Giemsa staining. Concordance was favorable for M. perstans although lower than for L. loa. Field's stain offers a rapid alternative to Giemsa stain for detection of L. loa microfilariae in thick blood smears. This could help accelerate diagnostics of blood filarial pathogens in mass screening programs or resource constrained health care institutions with high patient load.
Topics: Animals; Humans; Azure Stains; Loiasis; Microfilariae; Gabon; Filariasis; Coloring Agents; Loa
PubMed: 35895407
DOI: 10.4269/ajtmh.22-0061 -
Tropical Medicine & International... Aug 2022Loa loa and Mansonella perstans are two very common filarial species in Africa. Although microscopy is the traditional diagnostic method for human filariasis, several...
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.
Topics: Animals; Humans; Loa; Loiasis; Mansonella; Mansonelliasis; Polymerase Chain Reaction
PubMed: 35653502
DOI: 10.1111/tmi.13786 -
Diagnostics (Basel, Switzerland) Apr 2022Loiasis, caused by the filarial nematode , is endemic in Central and West Africa. has been associated with severe adverse reactions in high -infected individuals...
Loiasis, caused by the filarial nematode , is endemic in Central and West Africa. has been associated with severe adverse reactions in high -infected individuals receiving ivermectin during mass drug administration programs for the control of onchocerciasis and lymphatic filariasis. Diagnosis of loiasis still depends on microscopy in blood samples, but this is not effective for large-scale surveys. New diagnostics methods for loiasis are urgently needed. Previously, we developed a colorimetric high-sensitive and species-specific LAMP for DNA detection. Here, we evaluate it in a set of 100 field-collected clinical samples stored as dried blood spots. In addition, -LAMP was also evaluated in real-time testing and compared with microscopy and a specific PCR/nested PCR. A simple saponin/Chelex-based method was used to extract DNA. Colorimetric and real-time LAMP assays detected more samples with microscopy-confirmed and / mixed infections than PCR/nested-PCR. Samples with the highest microfilariae counts were amplified faster in real-time LAMP assays. Our -LAMP could be a promising molecular tool for the easy, rapid and accurate screening of patients for loiasis in endemic areas with low-resource settings. The real-time testing (feasible in a handheld device) could be very useful to rule out high-microfilariae loads in infected patients.
PubMed: 35626235
DOI: 10.3390/diagnostics12051079 -
PLoS Neglected Tropical Diseases Apr 2022Malaria, blood-borne filarial worms and intestinal parasites are all endemic in Gabon. This geographical co-distribution leads to polyparasitism and, consequently, the...
Malaria, blood-borne filarial worms and intestinal parasites are all endemic in Gabon. This geographical co-distribution leads to polyparasitism and, consequently, the possibility of immune-mediated interactions among different parasite species. Intestinal protozoa and helminths could modulate antimalarial immunity, for example, thereby potentially increasing or reducing susceptibility to malaria. The aim of the study was to compare the cytokine levels and cytokine ratios according to parasitic profiles of the population to determine the potential role of co-endemic parasites in the malaria susceptibility of populations. Blood and stool samples were collected during cross-sectional surveys in five provinces of Gabon. Parasitological diagnosis was performed to detect plasmodial parasites, Loa loa, Mansonella perstans, intestinal helminths (STHs) and protozoan parasites. Nested PCR was used to detect submicroscopic plasmodial infection in individuals with negative blood smears. A cytometric bead array was used to quantify interleukin (IL)-6, IL-10 and tumour necrosis factor (TNF)-α in the plasma of subjects with different parasitological profiles. Median IL-6 and IL-10 levels and the median IL-10/TNF-α ratio were all significantly higher among individuals with Plasmodium (P.) falciparum infection than among other participants (p<0.0001). The median TNF-α level and IL-10/IL-6 ratio were higher in subjects with STHs (p = 0.09) and P. falciparum-intestinal protozoa co-infection (p = 0.04), respectively. IL-6 (r = -0.37; P<0.01) and IL-10 (r = -0.37; P<0.01) levels and the IL-10/TNF-α ratio (r = -0.36; P<0.01) correlated negatively with age. Among children under five years old, the IL-10/TNF-α and IL-10/IL-6 ratios were higher in those with intestinal protozoan infections than in uninfected children. The IL-10/TNF-α ratio was also higher in children aged 5-15 years and in adults harbouring blood-borne filariae than in their control counterparts, whereas the IL-10/IL-6 ratio was lower in those aged 5-15 years with filariae and intestinal parasites but higher in adults with intestinal parasitic infections. Asymptomatic malaria is associated with a strong polarization towards a regulatory immune response, presenting high circulating levels of IL-10. P. falciparum/intestinal protozoa co-infections were associated with an enhanced IL-10 response. Immunity against malaria could differ according to age and carriage of other parasites. Helminths and intestinal protozoa can play a role in the high susceptibility to malaria currently observed in some areas of Gabon, but further investigations are necessary.
Topics: Animals; Child, Preschool; Cities; Coinfection; Cross-Sectional Studies; Cytokines; Gabon; Humans; Interleukins; Malaria; Malaria, Falciparum; Plasmodium falciparum; Rural Population; Tumor Necrosis Factor-alpha
PubMed: 35421083
DOI: 10.1371/journal.pntd.0010308 -
Journal of Travel Medicine Nov 2022Infection with Mansonella perstans is a neglected filariasis, widely distributed in sub-Saharan Africa, characterized by an elusive clinical picture; treatment for...
BACKGROUND
Infection with Mansonella perstans is a neglected filariasis, widely distributed in sub-Saharan Africa, characterized by an elusive clinical picture; treatment for mansonellosis is not standardized. This retrospective study aimed to describe the clinical features, treatment schemes and evolution, of a large cohort of imported cases of M. perstans infection seen in four European centres for tropical diseases.
METHODS
Mansonella perstans infections, diagnosed by identification of blood microfilariae in migrants, expatriates and travellers, collected between 1994 and 2018, were retrospectively analysed. Data concerning demographics, clinical history and laboratory examinations at diagnosis and at follow-up time points were retrieved.
RESULTS
A total of 392 patients were included in the study. Of the 281 patients for whom information on symptoms could be retrieved, 150 (53.4%) reported symptoms, abdominal pain and itching being the most frequent. Positive serology and eosinophilia were present in 84.4% and 66.1%, respectively, of those patients for whom these data were available. Concomitant parasitic infections were reported in 23.5% of patients. Treatment, administered to 325 patients (82.9%), was extremely heterogeneous between and within centres; the most commonly used regimen was mebendazole 100 mg twice a day for 1 month. A total of 256 (65.3%) patients attended a first follow-up, median 3 months (interquartile range 2-12) after the first visit; 83.1% of patients having received treatment based on mebendazole and/or doxycycline, targeting Wolbachia, became amicrofilaremic, 41.1-78.4% of whom within 12 months from single treatment.
CONCLUSIONS
Lack of specific symptoms, together with the inconstant positivity of parasitological and antibody-based assays in the infected population, makes the clinical suspicion and screening for mansonellosis particularly difficult. Prospective studies evaluating prevalence of infection in migrants from endemic areas, infection-specific morbidity, presence of Wolbachia endosymbionts in M. perstans populations from different geographical areas and efficacy of treatment regimens are absolutely needed to optimize the clinical management of infection.
Topics: Animals; Humans; Mansonella; Mansonelliasis; Retrospective Studies; Travel; Mebendazole; Prospective Studies; Travel-Related Illness; Wolbachia
PubMed: 35417002
DOI: 10.1093/jtm/taac048 -
PLoS Neglected Tropical Diseases Mar 2022Onchocercidae nematodes are heteroxenous parasites with worldwide distribution, and some of the species associated to animals may present zoonotic potential. Climatic...
Onchocercidae nematodes are heteroxenous parasites with worldwide distribution, and some of the species associated to animals may present zoonotic potential. Climatic changes and anthropic influences on the environment may result in vectors' proliferation, facilitating the spillover to humans and/or non-typical animal hosts. The Iguaçu National Park (PARNA Iguaçu), one of the most important Brazilian natural remanescents of Atlantic rainforest, is strongly affected by human activities such as tourism and agriculture. The complexity of this area is especially characterized by the close nexus between the rich wildlife, humans, and domestic animals, especially domestic dogs. Based on this, this research aimed to diagnose the Onchocercidae nematodes in wild carnivores and domestic dogs in the PARNA Iguaçu and the surrounding areas. For this, we collected 162 samples of seven species of wild carnivores and 225 samples of domestic dogs. The presence of microfilariae in the blood samples was diagnosed by the modified Knott's test and molecular screening, and the specific identification was based on sequencing of the myoHC and hsp70 genes. Microfilariae were detected only in ring-tailed coatis, in which we found five species: Mansonella sp. 1, Mansonela sp. 2, Onchocercidade gen. sp. 1, Onchocercidade gen. sp. 2, and Dirofilaria immitis. The morphological analysis supported the molecular findings. The domestic dogs were parasitized by Acanthocheilonema reconditum, representing a new locality record for this species. Phylogenetic analysis showed high genetic similarity among the four undetermined species and Mansonella spp., Brugia spp., and Wuchereria bancrofti. The presence of D. immitis in ring-tailed coatis may be result of spillover from dogs, even though the parasite was not diagnosed in the sampled dogs. The presence of several undetermined Onchocercidae species indicates the necessity of continuous investigations on wild and domestic animals from Neotropical area, especially considering the growing anthropic influence on forest remnants.
Topics: Animals; Brazil; Dirofilaria immitis; Dirofilariasis; Dog Diseases; Dogs; Forests; Microfilariae; Phylogeny; Spirurida
PubMed: 35259155
DOI: 10.1371/journal.pntd.0010213 -
PLoS Neglected Tropical Diseases Jan 2022Regular and comprehensive epidemiological surveys of the filarial nematodes Mansonella perstans and Loa loa in children, adolescents and adults living across Bioko...
Characterising co-infections with Plasmodium spp., Mansonella perstans or Loa loa in asymptomatic children, adults and elderly people living on Bioko Island using nucleic acids extracted from malaria rapid diagnostic tests.
BACKGROUND
Regular and comprehensive epidemiological surveys of the filarial nematodes Mansonella perstans and Loa loa in children, adolescents and adults living across Bioko Island, Equatorial Guinea are lacking. We aimed to demonstrate that blood retained on malaria rapid diagnostic tests, commonly deployed for malaria surveys, could be used as a source of nucleic acids for molecular based detection of M. perstans and L. loa. We wanted to determine the positivity rate and distribution of filarial nematodes across different age groups and geographical areas as well as to understand level of co-infections with malaria in an asymptomatic population.
METHODOLOGY
M. perstans, L. loa and Plasmodium spp. parasites were monitored by qPCR in a cross-sectional study using DNA extracted from a subset malaria rapid diagnostic tests (mRDTs) collected during the annual malaria indicator survey conducted on Bioko Island in 2018.
PRINCIPAL FINDINGS
We identified DNA specific for the two filarial nematodes investigated among 8.2% (263) of the 3214 RDTs screened. Positivity rates of M. perstans and L. loa were 6.6% and 1.5%, respectively. M. perstans infection were more prominent in male (10.5%) compared to female (3.9%) survey participants. M. perstans parasite density and positivity rate was higher among older people and the population living in rural areas. The socio-economic status of participants strongly influenced the infection rate with people belonging to the lowest socio-economic quintile more than 3 and 5 times more likely to be L. loa and M. perstans infected, respectively. No increased risk of being co-infected with Plasmodium spp. parasites was observed among the different age groups.
CONCLUSIONS/SIGNIFICANCE
We found otherwise asymptomatic individuals were infected with M. perstans and L. loa. Our study demonstrates that employing mRDTs probed with blood for malaria testing represents a promising, future tool to preserve and ship NAs at room temperature to laboratories for molecular, high-throughput diagnosis and genotyping of blood-dwelling nematode filarial infections. Using this approach, asymptomatic populations can be reached and surveyed for infectious diseases beyond malaria.
Topics: Adolescent; Adult; Animals; Child; Coinfection; Cross-Sectional Studies; DNA, Helminth; Equatorial Guinea; Female; Humans; Loa; Loiasis; Malaria; Male; Mansonella; Mansonelliasis; Middle Aged; Plasmodium; Prevalence; Socioeconomic Factors
PubMed: 35100277
DOI: 10.1371/journal.pntd.0009798 -
Infection, Genetics and Evolution :... Mar 2022To assess the emergent zoonotic disease risk posed by the voracious human-biting blackfly species Simulium oyapockense in the peripheral regions of an expanding urban...
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.
Topics: Animals; Insect Vectors; Mansonella; Mansonelliasis; Simuliidae; Zoonoses
PubMed: 34990852
DOI: 10.1016/j.meegid.2021.105200 -
Animals : An Open Access Journal From... Nov 2021Wild chimpanzee populations in West Africa () have dramatically decreased as a direct consequence of anthropogenic activities and infectious diseases. Little information...
Wild chimpanzee populations in West Africa () have dramatically decreased as a direct consequence of anthropogenic activities and infectious diseases. Little information is currently available on the epidemiology, pathogenic significance, and zoonotic potential of protist species in wild chimpanzees. This study investigates the occurrence and genetic diversity of intestinal and blood protists as well as filariae in faecal samples ( = 234) from wild chimpanzees in the Dindefelo Community Nature Reserve, Senegal. PCR-based results revealed the presence of intestinal potential pathogens ( spp.: 11.5%; : 2.1%; : 0.9%), protist of uncertain pathogenicity ( sp.: 5.6%), and commensal species (: 18.4%; : 5.6%). , , and were undetected. Blood protists including (0.4%), (1.3%), and (9.8%) were also identified. Sanger sequencing analyses revealed host-adapted genetic variants within , but other parasitic pathogens (, , , ) have zoonotic potential, suggesting that cross-species transmission between wild chimpanzees and humans is possible in areas where both species overlap. Additionally, we explored potential interactions between intestinal/blood protist species and seasonality and climate variables. Chimpanzees seem to play a more complex role on the epidemiology of pathogenic and commensal protist and nematode species than initially anticipated.
PubMed: 34828022
DOI: 10.3390/ani11113291