Possible transmission of Strongyloides fuelleborni between working Southern pig-tailed macaques (Macaca nemestrina) and their owners in Southern Thailand: Molecular identification and diversity.

Human strongyloidiasis is caused by Strongyloides stercoralis, S. fuelleborni fuelleborni and Strongyloides f. kellyi. Strongyloides fuelleborni is a soil-transmitted nematode parasite typically infecting non-human primates. The southern pig-tailed macaque (Macaca nemestrina) is distributed throughout the southern part of Thailand and could be a source of zoonotic transmission of this nematode. Here, we extracted DNA from Strongyloides speciescultured from the feces of southern pig-tailed macaques and their owners. Using PCR and sequencing of the extracted DNA, we compared the nucleotide sequences of these worms using portions of the 18S rDNA hypervariable region IV (HVR-IV) and the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene. Sequences from the 18S rRNA gene were obtained from worms from 23 southern pig-tailed macaques and from one owner. These sequences were identical with each other and with all East and Southeast Asian S. fuelleborni sequences (from Japan, Thailand, and Lao PDR) in the GenBank database. A median-joining network of published cox1 sequences (n = 123), in combination with the present 24 new sequences, represented 107 haplotypes distributed among six clusters, which corresponded to geographical localities but did not relate to host species. The S. fuelleborni cox1 sequences from some southern pig-tailed macaques and the one infected owner shared the same cox1 haplotype. This is the first evidence of likely zoonotic transmission of S. fuelleborni from a reservoir host, M. nemestrina.

Machine learning-based analyses support the existence of species complexes for Strongyloides fuelleborni and Strongyloides stercoralis.

Human strongyloidiasis is a serious disease mostly attributable to Strongyloides stercoralis and to a lesser extent Strongyloides fuelleborni, a parasite mainly of non-human primates. The role of animals as reservoirs of human-infecting Strongyloides is ill-defined, and whether dogs are a source of human infection is debated. Published multi-locus sequence typing (MLST) studies attempt to elucidate relationships between Strongyloides genotypes, hosts, and distributions, but typically examine relatively few worms, making it difficult to identify population-level trends. Combining MLST data from multiple studies is often impractical because they examine different combinations of loci, eliminating phylogeny as a means of examining these data collectively unless hundreds of specimens are excluded. A recently-described machine learning approach that facilitates clustering of MLST data may offer a solution, even for datasets that include specimens sequenced at different combinations of loci. By clustering various MLST datasets as one using this procedure, we sought to uncover associations among genotype, geography, and hosts that remained elusive when examining datasets individually. Multiple datasets comprising hundreds of S. stercoralis and S. fuelleborni individuals were combined and clustered. Our results suggest that the commonly proposed 'two lineage' population structure of S. stercoralis (where lineage A infects humans and dogs, lineage B only dogs) is an over-simplification. Instead, S. stercoralis seemingly represents a species complex, including two distinct populations over-represented in dogs, and other populations vastly more common in humans. A distinction between African and Asian S. fuelleborni is also supported here, emphasizing the need for further resolving these taxonomic relationships through modern investigations.

The Strongyloides stercoralis-hookworms association as a path to the estimation of the global burden of strongyloidiasis: A systematic review.

TRIAL REGISTRATION: PROSPERO (registration code CRD42019131127).

Update on the major imported helminth infections in travelers and migrants.

Helminth infections cause considerable morbidity worldwide and may be frequently underdiagnosed especially in areas of lower endemicity. Patients may harbor latent infections that may become symptomatic years or decades after the initial exposure and timely diagnosis may be critical to prevent complications and improve outcomes. In this context, disease in special populations, such as immunosuppressed patients, may be of particular concern. Heightened awareness and recent diagnostic developments may contribute to the correct management of helminth infections in nonendemic regions. A review of the main helminth infections in travelers and migrants (strongyloidiasis, taeniasis-neurocysticercosis and schistosomiasis) is presented, focusing on epidemiology, developments in diagnosis, treatment and prevention.

The effect of maternal Strongyloides venezuelensis infection on mice offspring susceptibility and immune response.

Species of Strongyloides infect a wide range of hosts worldwide. Due to their complex life cycle, it is hard to control the transmission of these parasites. Several species show evidence of vertical transmission; however, the impact of this transmission route on the susceptibility of the offspring has been poorly investigated. Herein, we used Strongyloides venezuelensis infected mice to evaluate transplacental and transmammary parasite transmission and their effect on the susceptibility of offspring. Swiss female mice were infected at the end of the gestation or during the breastfeeding period, and their offspring were examined for the presence of the parasite one week after infection of the mother. Our data showed that female mice infected with S. venezuelensis during gestation did not transmit the parasite to their offspring. On the other hand, all newborn mice breastfeeding in S. venezuelensis infected females got infected. To evaluate the effect of early exposure to the parasite on susceptibility and immune response of the hosts, the offspring of each experimental group (non-infected, gestation-infected, and breastfeeding-infected mothers) received anti-helminth treatment after parasite evaluation and were subcutaneously infected with S. venezuelensis upon reaching adulthood. Mice from the group of breastfeeding-infected mothers showed lower susceptibility to S. venezuelensis in adulthood in comparison with mice from non-infected mothers. The low parasite burden was accompanied by earlier eosinophil and neutrophil activation in the gut and higher serum levels of IgE. In contrast, S. venezuelensis infection in adult mice born from gestation-infected mothers presented with more worms in the intestine and lower levels of parasite-reactive IgM in serum in comparison with mice born from non-infected mothers, thus suggesting that early exposure to parasite antigens may modulate the protective immune response. Altogether, our data confirmed transmammary, but not transplacental, transmission of S. venezuelensis in mice and demonstrated that early exposure to the parasite and/or their antigens has an important effect on host susceptibility to a later infection.

Dogs are reservoir hosts for possible transmission of human strongyloidiasis in Thailand: molecular identification and genetic diversity of causative parasite species.

Human strongyloidiasis is a deleterious gastrointestinal disease mainly caused by Strongyloides stercoralis infection. We aimed to study the possible transmission of S. stercoralis between humans and pet animals. We isolated Strongyloides from humans and domestic dogs in the same rural community in north-east Thailand and compared the nucleotide sequences of derived worms using portions of the mitochondrial cytochrome c oxidase subunit 1 (cox1) and 18S ribosomal RNA (18S rRNA) genes. Twenty-eight sequences from the 18S rRNA gene were obtained from worms derived from humans (n = 23) and dogs (n = 5), and were identical with S. stercoralis sequences (from Thailand, Cambodia, Lao PDR and Myanmar) published in the GenBank database. The 28 cox1 sequences from humans and dogs showed high similarity to each other. The available published cox1 sequences (n = 150), in combination with our 28 sequences, represented 68 haplotypes distributed among four clusters. The 28 samples from the present study represented eight haplotypes including four new haplotypes. Dogs and humans shared the same haplotypes, suggesting the possibility of zoonotic transmission from pet dogs to humans. This is of concern since dogs and humans live in close association with each other.

From the feces to the genome: a guideline for the isolation and preservation of Strongyloides stercoralis in the field for genetic and genomic analysis of individual worms.

Strongyloidiasis is a soil-borne helminthiasis, which, in spite of the up to 370 million people currently estimated to be infected with its causing agent, the nematode Strongyloides stercoralis, is frequently overlooked. Recent molecular taxonomic studies conducted in Southeast Asia and Australia, showed that dogs can carry the same genotypes of S. stercoralis that also infect humans, in addition to a presumably dog-specific Strongyloides species. This suggests a potential for zoonotic transmission of S. stercoralis from dogs to humans. Although natural S. stercoralis infections have not been reported in any host other than humans, non-human primates and dogs, other as yet unidentified animal reservoirs cannot be excluded. Molecular studies also showed that humans carry rather different genotypes of S. stercoralis. As a result, their taxonomic status and the question of whether they differ in their pathogenic potential remains open. It would therefore be very important to obtain molecular genetic/genomic information about S. stercoralis populations from around the world. One way of achieving this (with little additional sampling effort) would be that people encountering S. stercoralis in the process of their diagnostic work preserve some specimens for molecular analysis. Here we provide a guideline for the isolation, preservation, genotyping at the nuclear 18S rDNA and the mitochondrial cox1 loci, and for whole genome sequencing of single S. stercoralis worms. Since in many cases the full analysis is not possible or desired at the place and time where S. stercoralis are found, we emphasize when and how samples can be preserved, stored and shipped for later analysis. We hope this will benefit and encourage researchers conducting field studies or diagnostics to collect and preserve S. stercoralis for molecular genetic/genomic analyses and either analyze them themselves or make them available to others for further analysis.

Assessment of the global paradigms of genetic variability in Strongyloides stercoralis infrapopulations determined by mitochondrial DNA sequences.

Microevolutionary data of Strongyloides stercoralis infrapopulations are regarded as a requirement for determining the global genetic structure and transmission paradigms of this neglected tropical nematode among the neighboring countries of the world. English databases were searched from 2010 to 2019, analyzing a total of 10 publications. The cytochrome c oxidase subunit 1 sequences of S. stercoralis isolated from Asian and African continents were subjected to calculate the diversity indices and genetic differentiation. A parsimonious haplotype network indicated a star-like trait a total of 106 (Homo sapiens) and 48 haplotypes (Canid) being grouped into four distinct geographical haplogroups. A significant genetic diversity was identified in human-derived S. stercoralis (Haplotype diversity: 0.78) and those with dog (Hd: 0.86) origins. Cladistic phylogenetic tree indicated the Japanese, Thailandish, and Myanmarese clades have a sister relationship with the Laotian clade. The statistically significant Fst values indicated that human S. stercoralis populations of Japanese-Thailandish, Japanese-Myanmarese, and Japanese-Laotian origins were genetically differentiated (Fst: 0.48430 to 0.54903). We conclude that a high gene migration of human strongyloidiasis is being unequivocally shared between the Laotion-Myanmarese and Laotion-Thailandish population pairs. The current findings enhance our knowledge to assess the transmission dynamics and the evolutionary patterns of S. stercoralis in various geographical regions of the globe; also it will serve as a basis for public health policy to control human strongyloidiasis particularly in immunocompromised individuals. Besides, the infected canids and other environmental reservoirs for zoonotic transmission of S. stercoralis to humans should be de-wormed along with their owners.

Detection of classic and cryptic Strongyloides genotypes by deep amplicon sequencing: A preliminary survey of dog and human specimens collected from remote Australian communities.

Strongyloidiasis is caused by the human infective nematodes Strongyloides stercoralis, Strongyloides fuelleborni subsp. fuelleborni and Strongyloides fuelleborni subsp. kellyi. The zoonotic potential of S. stercoralis and the potential role of dogs in the maintenance of strongyloidiasis transmission has been a topic of interest and discussion for many years. In Australia, strongyloidiasis is prevalent in remote socioeconomically disadvantaged communities in the north of the continent. Being an isolated continent that has been separated from other regions for a long geological period, description of diversity of Australian Strongyloides genotypes adds to our understanding of the genetic diversity within the genus. Using PCR and amplicon sequencing (Illumina sequencing technology), we sequenced the Strongyloides SSU rDNA hyper-variable I and hyper-variable IV regions using Strongyloides-specific primers, and a fragment of the mtDNA cox1 gene using primers that are broadly specific for Strongyloides sp. and hookworms. These loci were amplified from DNA extracted from Australian human and dog faeces, and one human sputum sample. Using this approach, we confirm for the first time that potentially zoonotic S. stercoralis populations are present in Australia, suggesting that dogs represent a potential reservoir of human strongyloidiasis in remote Australian communities.

Prevalence and risk factors of Strongyloides stercoralis infection among Orang Asli schoolchildren: new insights into the epidemiology, transmission and diagnosis of strongyloidiasis in Malaysia.

This cross-sectional study aimed to determine the prevalence and risk factors of S. stercoralis infection among 1142 Orang Asli primary schoolchildren in six different states of Peninsular Malaysia. Fecal samples were examined using direct smear, formalin-ether sedimentation (FES), agar plate culture (APC) and PCR techniques. Overall, 15.8% of the children were found to be infected with S. stercoralis. The prevalence was 0.2, 1.3, 15.2 and 13.7% by direct smear, FES, APC and PCR, respectively. Multivariate analysis showed that an age of >10 years, being male, belonging to a Proto-Malay tribe, belonging to the Senoi tribe, indiscriminate defecation, using an unimproved water source for drinking water and not wearing shoes when outside were the significant risk factors of infection among these children. In conclusion, we provide new evidence on the occurrence of S. stercoralis in Malaysia to show that there is a relatively high prevalence of infection among Orang Asli schoolchildren. Therefore, the use of specific methods for detecting S. stercoralis should be considered when screening these children for intestinal parasites. Moreover, prevention and control measures specific to S. stercoralis should be integrated into the intestinal parasitic infections control programme in Malaysia.

Morbidity Associated with Chronic Strongyloides stercoralis Infection: A Systematic Review and Meta-Analysis.

Strongyloides stercoralis, a worldwide-distributed soil-transmitted helminth, causes chronic infection which may be life threatening. Limitations of diagnostic tests and nonspecificity of symptoms have hampered the estimation of the global morbidity due to strongyloidiasis. This work aimed at assessing S. stercoralis-associated morbidity through a systematic review and meta-analysis of the available literature. MEDLINE, Embase, CENTRAL, LILACS, and trial registries (WHO portal) were searched. The study quality was assessed using the Newcastle-Ottawa scale. Odds ratios (ORs) of the association between symptoms and infection status and frequency of infection-associated symptoms were calculated. Six articles from five countries, including 6,014 individuals, were included in the meta-analysis-three were of low quality, one of high quality, and two of very high quality. Abdominal pain (OR 1.74 [CI 1.07-2.94]), diarrhea (OR 1.66 [CI 1.09-2.55]), and urticaria (OR 1.73 [CI 1.22-2.44]) were associated with infection. In 17 eligible studies, these symptoms were reported by a large proportion of the individuals with strongyloidiasis-abdominal pain by 53.1% individuals, diarrhea by 41.6%, and urticaria by 27.8%. After removing the low-quality studies, urticaria remained the only symptom significantly associated with S. stercoralis infection (OR 1.42 [CI 1.24-1.61]). Limitations of evidence included the low number and quality of studies. Our findings especially highlight the appalling knowledge gap about clinical manifestations of this common yet neglected soil-transmitted helminthiasis. Further studies focusing on morbidity and risk factors for dissemination and mortality due to strongyloidiasis are absolutely needed to quantify the burden of S. stercoralis infection and inform public health policies.

Implementation of a Strongyloides screening strategy in solid organ transplant donors and recipients.

BACKGROUND: Strongyloides stercoralis infects 100 million people worldwide. Mortality rates in hyperinfection syndrome exceed 50%. Donor-derived Strongyloides infection has occurred after heart, kidney, kidney-pancreas and liver transplantation; yet, only 10% of the US organ procurement organizations currently screen donors for strongyloidiasis. METHODS: We report a fatal case of donor-derived disseminated Strongyloides infection in a liver transplant recipient. Following this case, we implemented universal screening and treatment of donors and recipients. We reviewed our local epidemiology and outcomes after protocol implementation. RESULTS: From a total of 355 deceased donors accepted at our center between January 2016, and March 2018, 14 (3.9%) had positive Strongyloides serology. Except for the index case, all other recipients of Strongyloides antibody-positive donors within that period (including 10 kidneys, 3 livers, one combined liver/kidney, and one kidney/pancreas from eight seropositive donors) received post-transplant prophylaxis with ivermectin, and to date are alive and doing well without signs of infection. Between October 2015, and September 2016, a total of 441 deceased donor solid organ transplants were performed at our center. 220 of these recipients had pretransplant Strongyloides serology available, and 23 of them were seropositive (10.5%). Within the first two years after the implementation of universal screening and treatment of donors and recipients, we had no cases of Strongyloides reactivation in our center. CONCLUSIONS: Implementation of a Strongyloides screening and treatment protocol in our center was an effective strategy to prevent both recipient- and donor-derived strongyloidiasis. Transplant centers should consider implementation of Strongyloides preventive strategies.

Donor-derived strongyloidiasis in a Saudi pediatric kidney transplant recipient: A case report and mini-review.

S. stercoralis infection has been identified as a donor-derived infection in cases of solid organ transplant among recipients with no prior risk factor for parasitic exposure. Worldwide and regional reports from the adult kidney transplant population highlight this indirect method of infection and caution about delayed diagnosis, severe complications, and death related to donor-derived S. stercoralis infection. We report a deceased-donor-derived S. stercoralis infection in a 12-year-old Saudi girl who underwent kidney transplantation. This is the first pediatric case reported outside the United States of America. Although she presented with mild bouts of gastrointestinal symptoms, the need for additional immune suppression put her at risk of serious complications. A literature review highlights the importance of awareness about S. stercoralis infections and complications in kidney transplant recipients, pretransplant screening of donors based on risk assessment, and the challenges with treatment availability and duration in this vulnerable population.

Donor-derived strongyloidiasis after organ transplantation in Norway.

Strongyloides stercoralis is an intestinal helminth which in humans can cause asymptomatic chronic infection maintained for decades through its auto-infective cycle. During solid organ transplantation, recipients may unintentionally receive an organ infected with strongyloides. This is a very rare complication but may have deadly outcome if not detected. We hereby report two transplant recipients whom developed Strongyloides hyperinfection syndrome after organ transplantation from the same deceased donor. Recipient 1 was kidney transplanted and presented at day 65 post engraftment with diarrhea and subsequent septicemia and gastric retention. Larvae were detected in gastric aspirate. Recipient 2 was simultaneously kidney and pancreas transplanted and presented at day 90 post engraftment also with gastric retention and septicemia. Larvae were demonstrated on duodenal biopsy and stool sample. The clinical course was complicated with severe duodenal bleedings, gastric retention, meningitis, and prolonged hospitalization. Retrospective testing of pre-transplant donor serum was positive for Strongyloides stercoralis antibodies. As a result of disease severity and gastric retention albenazole was administered via a jejunal tube and ivermectin subcutaneously in both recipients. S stercoralis was successfully eradicated and the transplants ended up with unaffected graft function. Following these two cases, we started systematic screening of all deceased donors for serum Strongyloides IgG in October 2016. After having screened 150 utilized donors one tested positive for Strongyloides, which initiated prophylactic ivermectin treatment to organ recipients. No symptoms or disease developed. Our center will continue to screen all donors as prophylactic treatment may avert this potentially lethal complication in cases of donor-derived Strongyloides infection.

Seroepidemiological aspects of human infection by Strongyloides stercoralis in Alfenas, southern Minas Gerais, Brazil.

INTRODUCTION: In most Strongyloides stercoralis infected individuals, nematoidosis occurs asymptomatically, but in immunocompromised patients, it can cause hyperinfection. Serological techniques seem to be a good alternative for detecting this parasite. METHODS: The frequency of seropositivity for strongyloidiasis in Alfenas, MG, was estimated using the enzyme linked immunosorbent assay on blood samples, between May and August of 2015. RESULTS: Out of 258 samples tested, 53.9% were positive, and the frequency of seropositive individuals was higher in the peripheral districts of the municipality. CONCLUSIONS: The results indicate high seropositivity rates for strongyloidiasis among the residents of Alfenas city.

Seroepidemiological aspects of human infection by Strongyloides stercoralis in Alfenas, southern Minas Gerais, Brazil

Abstract INTRODUCTION: In most Strongyloides stercoralis infected individuals, nematoidosis occurs asymptomatically, but in immunocompromised patients, it can cause hyperinfection. Serological techniques seem to be a good alternative for detecting this parasite. METHODS The frequency of seropositivity for strongyloidiasis in Alfenas, MG, was estimated using the enzyme linked immunosorbent assay on blood samples, between May and August of 2015. RESULTS: Out of 258 samples tested, 53.9% were positive, and the frequency of seropositive individuals was higher in the peripheral districts of the municipality. CONCLUSIONS: The results indicate high seropositivity rates for strongyloidiasis among the residents of Alfenas city.

MaxEnt modeling of soil-transmitted helminth infection distributions in Thailand.

Infections due to soil-transmitted helminths (STHs), i.e. Ascaris lumbricoides, Trichuris trichiura, hookworms, and Strongyloides stercoralis, are widely distributed in tropical and subtropical areas in which approximately 1.5 billion people are infected. A clear understanding of the epidemiology and distribution of diseases is an important aid for control and prevention. The aim of our study was to identify the effects of environmental and climatic factors on distribution patterns of STHs and to develop a risk map for STH infections under current environmental and climate regimes in Thailand. Geographical information systems (GIS), remote sensing, and Maximum Entropy (MaxEnt) algorithm software were used to determine the significant factors and to create predictive risk maps for STH infections in Thailand. The disease data from Thailand covered the years from 1969 to 2014, while environmental and climatic data were compiled from the Worldclim database, MODIS satellite imagery, Soilgrids and ISCGM. The models predicted that STHs occur mainly in southern Thailand. Mean annual precipitation was the factor most affecting the current distribution of A. lumbricoides, T. trichiura, and S. stercoralis. Land cover class was the main predictor for distribution of S. stercoralis and important for hookworms. Altitude was the dominant factor affecting the distribution of hookworms, and mean temperature of the wettest quarter was significantly associated with A. lumbricoides distribution. A predicted distribution map of STHs to identify environmental risk factors in Thailand is presented. This work provides a model for use in STH monitoring and health planning not only in Thailand but also in other countries with similar disease conditions.

Prevalence of Strongyloides stercoralis infection and other soil-transmitted helminths by cross-sectional survey in a rural community in Gisagara District, Southern Province, Rwanda.

Background: Strongyloides stercoralis is one of the most neglected tropical diseases. Sparse, dated central African and Rwandan data on seroprevalence are available to guide public health efforts and clinical care. Methods: In February 2016 we conducted a community-based cross-sectional study among 539 asymptomatic participants in a rural area in the Gisagara District, Southern Province, Rwanda. Direct faecal smear (DFS) and modified Koga agar plate culture (APC) were used to detect S. stercoralis infection in a single stool sample. Data on other soil-transmitted helminths diagnosed by DFS were also recorded. Results: Four intestinal helminth infections were diagnosed, with S. stercoralis (17.4%) and hookworms (8.2%) seen most often. APC, compared with DFS, increased the diagnosis rate for S. stercoralis from 1.9% to 17.4% (p<0.01). The prevalence was higher in farmers and those with lower socio-economic status. Females were less often infected than males (odds ratio 0.6 [95% confidence interval 0.3 to 0.9], p=0.02). Conclusions: S. stercoralis is highly prevalent among the general population in a rural area of Gisagara District, Southern Province, Rwanda. Access to effective diagnosis and treatment is needed for this neglected disease.

Fecal parasite risk in the endangered proboscis monkey is higher in an anthropogenically managed forest environment compared to a riparian rain forest in Sabah, Borneo.

Understanding determinants shaping infection risk of endangered wildlife is a major topic in conservation medicine. The proboscis monkey, Nasalis larvatus, an endemic primate flagship species for conservation in Borneo, is endangered through habitat loss, but can still be found in riparian lowland and mangrove forests, and in some protected areas. To assess socioecological and anthropogenic influence on intestinal helminth infections in N. larvatus, 724 fecal samples of harem and bachelor groups, varying in size and the number of juveniles, were collected between June and October 2012 from two study sites in Malaysian Borneo: 634 samples were obtained from groups inhabiting the Lower Kinabatangan Wildlife Sanctuary (LKWS), 90 samples were collected from groups of the Labuk Bay Proboscis Monkey Sanctuary (LBPMS), where monkeys are fed on stationary feeding platforms. Parasite risk was quantified by intestinal helminth prevalence, host parasite species richness (PSR), and eggs per gram feces (epg). Generalized linear mixed effect models were applied to explore whether study site, group type, group size, the number of juveniles per group, and sampling month predict parasite risk. At the LBPMS, prevalence and epg of Trichuris spp., strongylids, and Strongyloides spp. but not Ascaris spp., as well as host PSR were significantly elevated. Only for Strongyloides spp., prevalence showed significant changes between months; at both sites, the beginning rainy season with increased precipitation was linked to higher prevalence, suggesting the external life cycle of Strongyloides spp. to benefit from humidity. Higher prevalence, epgs, and PSR within the LBPMS suggest that anthropogenic factors shape host infection risk more than socioecological factors, most likely via higher re-infection rates and chronic stress. Noninvasive measurement of fecal parasite stages is an important tool for assessing transmission dynamics and infection risks for endangered tropical wildlife. Findings will contribute to healthcare management in nature and in anthropogenically managed environments.