Application of Nested-qPCR-High Resolution Melting (HRM) Technology on Strongyloides stercoralis Isolates from Iran.

PURPOSE: Strongyloides stercoralis is a parasite with special characteristics presenting it as a unique nematode. Iran is an endemic area for S. stercoralis. In this study, nested-qPCR-high resolution melting (HRM) technology was applied on some human isolates of S. stercoralis from this country by focusing on evolutionary genetics analysis. METHODS: Twelve human isolates of S. stercoralis were collected from four endemic provinces of Iran. Genomic DNA was extracted from a single filariform larva for every isolate. Using specific primers targeting partial regions in cox1 gene, nested-qPCR-HRM was performed and melting-curve profiles were analyzed alongside the evaluation of genetic proximity and phylogenetic analysis using MEGA7 and DnaSP5 software. RESULTS: The melting temperature (Tm) values of the isolates were 77.9 °C-78.3 °C. All isolates from Guilan, Mazandaran, and Khouzestan Provinces shared Tm values of 78.2 °C to 78.3 °C, while the isolates from Hormozgan Province showed Tm values of 77.9 °C, 78.0 °C, and 78.1 °C. The phylogenetic tree illustrated that the sequences of the current study included nine haplotypes. Tajima's D index analyses showed that cox1 gene in S. stercoralis isolates was negative (Tajima's D = - 0.27). CONCLUSION: The isolates were divided into five temperature groups. Although HRM assay compared to PCR sequencing identified more limited genetic changes, it revealed that the mean of Tm of the isolates from Hormozgan Province was lower than those of other provinces and represented specific haplotypes for this geographical region on the phylogenetic tree.

Discovery and Structure-Activity Relationships of Novel ssDAF-12 Receptor Modulators.

The nuclear receptor ssDAF-12 has been recognized as the key molecular player regulating the life cycle of the nematode parasite Strongyloides stercoralis. ssDAF-12 ligands permit the receptor to function as an on/off switch modulating infection, making it vulnerable to therapeutic intervention. In this study, we report the design and synthesis of a set of novel dafachronic acid derivatives, which were used to outline the first structure-activity relationship targeting the ssDAF-12 receptor and to unveil hidden properties shared by the molecular shape of steroidal ligands that are relevant to the receptor binding and modulation. Moreover, biological results led to the discovery of sulfonamide 3 as a submicromolar ssDAF-12 agonist endowed with a high receptor selectivity, no toxicity, and improved properties, as well as to the identification of unprecedented ssDAF-12 antagonists that can be exploited in the search for novel chemical tools and alternative therapeutic approaches for treating parasitism such as Strongyloidiasis.

Improved diagnostic sensitivity of human strongyloidiasis using point-of-care mixed recombinant antigen-based immunochromatography.

Strongyloidiasis is a neglected tropical disease that can cause fatal complications due to hyperinfection and disseminated strongyloidiasis in immunocompromised patients. We used two Strongyloides stercoralis recombinant antigenic proteins, L3NieAg.01 (NIE) and IgG-immunoreactive antigen (SsIR), to develop the recombinant antigen-based immunochromatography test (ICT) kit. We constructed and compared kits using either the NIE (NIE ICT kit) or the SsIR (SsIR ICT kit) antigens and a kit using a mixture of both (NIE-SsIR ICT kit) for detection of anti-Strongyloides IgG antibody in human serum samples. Serum samples from normal healthy individuals (Group I, n = 40), proven strongyloidiasis patients (Group II, n = 100), and those with other parasitic infections (Group III, n = 154) were evaluated. Sensitivity and specificity were 81.0% and 84.0% for the NIE ICT kit, 89.0% and 83.5% for the SsIR ICT kit, and 95.0% and 90.2% for the NIE-SsIR ICT kit, respectively. The NIE-SsIR ICT kit provided the best diagnostic results; it can supplement stool examination for clinical diagnosis and can be used to screen for asymptomatic S. stercoralis infection in people at risk in endemic areas. The NIE-SsIR ICT kit can also be used in large-scale sero-epidemiological investigations in endemic areas without the need for additional facilities or ancillary supplies.

Title: Amélioration de la sensibilité diagnostique de l'anguillulose humaine grâce à l'immunochromatographie à base d'antigènes recombinants mixtes sur le lieu d'intervention. Abstract: La strongyloïdose est une maladie tropicale négligée qui peut entraîner des complications mortelles dues à une hyperinfection et à une strongyloïdose disséminée chez les patients immunodéprimés. Nous avons utilisé deux protéines antigéniques recombinantes de Strongyloides stercoralis, L3NieAg.01 (NIE) et l'antigène immunoréactif IgG (SsIR), pour développer un kit de test d'immunochromatographie (TIC) basé sur les antigènes recombinants. Nous avons construit et comparé des kits utilisant les antigènes NIE (kit NIE ICT), SsIR (kit SsIR ICT) ou un mélange des deux (kit NIE-SsIR ICT) pour la détection des anticorps IgG anti-Strongyloides dans des échantillons de sérum humain. Des échantillons de sérum provenant d'individus normaux en bonne santé (groupe I, n = 40), de patients atteints d'anguillulose avérée (groupe II, n = 100) et de patients atteints d'autres infections parasitaires (groupe III, n = 154) ont été évalués. La sensibilité et la spécificité étaient respectivement de 81,0 % et 84,0 % pour le kit NIE ICT, 89,0 % et 83,5 % pour le kit SsIR ICT, et 95,0 % et 90,2 % pour le kit NIE-SsIR ICT. Le kit NIE-SsIR ICT a fourni les meilleurs résultats de diagnostic ; il peut compléter l'examen des selles pour le diagnostic clinique et peut être utilisé pour dépister une infection asymptomatique à S. stercoralis chez les personnes à risque dans les zones d'endémie. Le kit NIE-SsIR ICT peut également être utilisé dans des enquêtes séro-épidémiologiques à grande échelle dans les zones endémiques sans nécessiter d'installations supplémentaires ou de fournitures auxiliaires.

Human strongyloidiasis: complexities and pathways forward.

Strongyloidiasis is a World Health Organization neglected tropical disease usually caused by Strongyloides stercoralis, a parasitic worm with a complex life cycle. Globally, 300-600 million people are infected through contact with fecally contaminated soil. An autoinfective component of the life cycle can lead to chronic infection that may be asymptomatic or cause long-term symptoms, including malnourishment in children. Low larval output can limit the sensitivity of detection in stool, with serology being effective but less sensitive in immunocompromise. Host immunosuppression can trigger catastrophic, fatal hyperinfection/dissemination, where large numbers of larvae pierce the bowel wall and disseminate throughout the organs. Stable disease is effectively treated by single-dose ivermectin, with disease in immunocompromised patients treated with multiple doses. Strategies for management include raising awareness, clarifying zoonotic potential, the development and use of effective diagnostic tests for epidemiological studies and individual diagnosis, and the implementation of treatment programs with research into therapeutic alternatives and medication safety.

Comparison of different PCR amplification targets for molecular diagnosis of Strongyloides stercoralis.

Molecular techniques are an alternative for the diagnosis of strongyloidiasis, produced by Strongyloides stercoralis. However, it is necessary to determine the best amplification target for the populations of this parasite present in a geographical area and standardize a polymerase chain reaction (PCR) protocol for its detection. The objectives of this work were the comparison of different PCR targets for molecular detection of S. stercoralis and the standardization of a PCR protocol for the selected target with the best diagnostic results. DNA extraction was performed from parasite larvae by saline precipitation. Three amplification targets of the genes encoding ribosomal RNA 18S (18S rDNA) and 5.8S (5.8S rDNA) and cytochrome oxidase 1 (COX1) of S. stercoralis were compared, and the PCR reaction conditions for the best target were standardized (concentration of reagents and template DNA, hybridization temperature, and number of cycles). The analytical sensitivity and specificity of the technique were determined. DNA extraction by saline precipitation made it possible to obtain DNA of high purity and integrity. The ideal target was the 5.8S rDNA, since the 18S rDNA yielded non-reproducible results and COX1 never amplified under any condition tested. The optimal conditions for the 5.8S rDNA-PCR were: 1.5 mM MgCl2, 100 µM dNTPs, 0.4 µM primers, and 0.75 U DNA polymerase, using 35 cycles and a hybridization temperature of 60 °C. The analytical sensitivity of the PCR was 1 attogram of DNA, and the specificity was 100%. Consequently, the 5.8S rDNA was shown to be highly sensitive and specific for the detection of S. stercoralis DNA.

Upper Gastrointestinal Bleeding Induced by Gastric Ulcer Secondary to Strongyloidiasis: A Case Report.

Strongyloidiasis, a parasitic infestation by Strongyloides stercoralis, involves the gastrointestinal tract with a spectrum from duodenitis to enterocolitis. However, gastric involvement with the manifestation of upper gastrointestinal bleeding is an extremely rare condition due to Strongyloides stercoralis. Due to irregular excretion of larvae, unclear symptoms, paucity of effective diagnostic tools and low parasitic load, makes clinicians difficult to reach the diagnosis of strongyloidiasis. Here, we present a case of upper gastrointestinal bleeding due to a large gastric ulcer whose aetiology was identified to be Strongyloides stercoralis infection of the gastric region by the diagnosis of exclusion. Keywords: gastric ulcer; gastrointestinal haemorrhage; Strongyloides stercoralis; strongyloidiasis.

Investigation of donor-derived Strongyloides stercoralis infection in multiple solid organ transplant recipients-California, Michigan, Ohio, 2022.

BACKGROUND: The Centers for Disease Control and Prevention led an investigation to determine if Strongyloides infection in a right kidney recipient was an existing chronic infection, or if the infection was transmitted from an infected organ donor. METHODS: Evidence regarding the organ donor and organ recipients Strongyloides testing, treatment, and risk factors were gathered and evaluated. The case classification algorithm created by the Disease Transmission Advisory Committee was utilized. RESULTS: The organ donor had risk factors for Strongyloides infection; the banked donor specimen, submitted for serology testing 112 days post-donor death, was positive. The right kidney recipient was negative for Strongyloides infection pretransplant. Strongyloides infection was diagnosed via small bowel and stomach biopsies. The left kidney recipient had risk factors for Strongyloides infection. Two posttransplant Strongyloides antibody tests were negative at 59 and 116 days posttransplant; repeat antibody tests returned positive at 158 and 190 days posttransplant. Examination of bronchial alveolar lavage fluid collected 110 days posttransplant from the heart recipient showed a parasite morphologically consistent with Strongyloides species. She subsequently developed complications from Strongyloides infection, including hyperinfection syndrome and disseminated strongyloidiasis. Based on the evidence from our investigation, donor-derived strongyloidiasis was suspected in one recipient and proven in two recipients. CONCLUSION: The results of this investigation support the importance of preventing donor-derived Strongyloides infections by laboratory-based serology testing of solid organ donors. Donor positive testing results would direct the monitoring and treatment of recipients to avoid severe complications.

From past to present: opportunities and trends in the molecular detection and diagnosis of Strongyloides stercoralis.

Strongyloides stercoralis is a soil-transmitted helminth that is mainly found in the tropical and subtropical regions and affects approximately 600 million people globally. The medical importance of strongyloidiasis lies in its capacity to remain asymptomatic and chronically unnoticed until the host is immunocompromised. Additionally, in severe strongyloidiasis, hyperinfection syndrome and larva dissemination to various organs can occur. Parasitological techniques such as Baermann-Moraes and agar plate culture to detect larvae in stool samples are the current gold standard. However, the sensitivity might be inadequate, especially with reduced worm burden. Complementing parasitological techniques, immunological techniques including immunoblot and immunosorbent assays are employed, with higher sensitivity. However, cross-reactivity to other parasites may occur, hampering the assay's specificity. Recently, advances in molecular techniques such as polymerase chain reaction and next-generation sequencing technology have provided the opportunity to detect parasite DNA in stool, blood, and environmental samples. Molecular techniques, known for their high sensitivity and specificity, have the potential to circumvent some of the challenges associated with chronicity and intermittent larval output for increased detection. Here, as S. stercoralis was recently included by the World Health Organization as another soil-transmitted helminth targeted for control from 2021 to 2030, we aimed to present a review of the current molecular techniques for detecting and diagnosing S. stercoralis in a bid to consolidate the molecular studies that have been performed. Upcoming molecular trends, especially next-generation sequencing technologies, are also discussed to increase the awareness of its potential for diagnosis and detection. Improved and novel detection methods can aid in making accurate and informed choices, especially in this era where infectious and non-infectious diseases are increasingly commonplace.

Modulation of circulating cytokine production in alcoholic patients infected with Strongyloides stercoralis.

Strongyloidiasis control is associated with a Th2 immune response. However, alcohol ingestion plays an important role in modulating the immune system. The aim of this study is to evaluate the occurrence of Strongyloides stercoralis infection in alcoholic patients, the levels of circulating cytokines (IFN-γ, IL-2, IL-4, IL-5, IL-10, IL-15 and IL-17), and its correlation with modulation of parasitic load in alcoholic individuals infected with S. stercoralis. A total of 336 alcoholic patients, treated at the Alcoholic Care and Treatment Center were included in this study. The cytokine levels were measured by a commercial ELISA in 80 sera divided into four groups with 20 individuals each: alcoholics infected (ASs+) and not infected (ASs-) with S. stercoralis and non-alcoholics infected (NASs+) and not infected (NASs-) with the helminth. S. stercoralis frequency in alcoholic patients was 16.1% (54/336). The parasitic load varied from 1 to 546 larvae/g of faeces, median and interquartile range (IQR) of 9 and 1.0-62.5 larvae/g of faeces, while in non-alcoholic individuals the parasitic load was less than 10 larvae/g of faeces. Levels of circulating IL-4 were significantly higher in ASs+ when compared with NASs- group (p < .05). An inverse correlation between serum levels of IFN-γ and parasitic load in alcoholic patients infected with S. stercoralis was observed (r = -601; p < 0.01). These results suggest that modulation of IFN-γ production occurs in alcoholic individuals with high parasitic burden.

Chronic Strongyloides stercoralis infection increases presence of the Ruminococcus torques group in the gut and alters the microbial proteome.

We explored the impact of chronic Strongyloides stercoralis infection on the gut microbiome and microbial activity in a longitudinal study. At baseline (time-point T0), 42 fecal samples from matched individuals (21 positive for strongyloidiasis and 21 negative) were subjected to microbiome 16S-rRNA sequencing. Those positive at T0 (untreated then because of COVID19 lockdowns) were retested one year later (T1). Persistent infection in these individuals indicated chronic strongyloidiasis: they were treated with ivermectin and retested four months later (T2). Fecal samples at T1 and T2 were subjected to 16S-rRNA sequencing and LC-MS/MS to determine microbial diversity and proteomes. No significant alteration of indices of gut microbial diversity was found in chronic strongyloidiasis. However, the Ruminococcus torques group was highly over-represented in chronic infection. Metaproteome data revealed enrichment of Ruminococcus torques mucin-degrader enzymes in infection, possibly influencing the ability of the host to expel parasites. Metaproteomics indicated an increase in carbohydrate metabolism and Bacteroidaceae accounted for this change in chronic infection. STITCH interaction networks explored highly expressed microbial proteins before treatment and short-chain fatty acids involved in the synthesis of acetate. In conclusion, our data indicate that chronic S. stercoralis infection increases Ruminococcus torques group and alters the microbial proteome.

Prevalence and risk factors of Strongyloides stercoralis in haemodialysis in Cochabamba, Bolivia: a cross-sectional study.

BACKGROUND: Strongyloidiasis is an infectious disease that can be fatal in immunocompromised patients. Patients with end-stage renal failure who are on dialysis have a considerably weakened immune system, and organ transplantation is a major risk factor for severe strongyloidiasis. Knowledge of the local epidemiology in tropical and subtropical areas is an essential prerequisite for designing an appropriate strategy to prevent this potentially lethal complication. In this study, we aimed to estimate the prevalence and associated risk factors of S. stercoralis infection in patients on dialysis in Cochabamba, Bolivia. METHODS: A cross-sectional study was carried out among patients undergoing haemodialysis in Cochabamba (elevation 2,500 m, temperate climate), collecting information on socio-demographic, lifestyle, and clinical variables, and using one coproparasitological technique (the modified Baermann technique) and one serological (ELISA) test for S.stercoralis diagnosis. RESULTS: In total, 149 patients participated in the study (mean age = 51.4 years, 48.3% male). End-stage renal disease was predominantly (59%) of hypertensive and/or diabetic origin. The positive serological prevalence was 18.8% (95% CI: 13.3%-25.9%). Based on the sensitivity and specificity of the ELISA test, the estimate of the actual prevalence was 15.1% (95% CI: 9.4%-20.7%). Stool samples of 105 patients (70.5%) showed a coproparasitological prevalence of 1.9% (95% CI: 0.52%-6.68%). No potential risk factors were significantly associated with S. stercoralis infection. CONCLUSIONS: We found a high seroprevalence of S. stercoralis in Bolivian patients undergoing haemodialysis in Cochabamba. We recommend presumptive antiparasitic treatment at regular intervals to avoid the potentially fatal complications of severe strongyloidiasis.

Larva Currens: Report of Seven Cases and Literature Review.

Strongyloidiasis is a frequent and often unrecognized parasitic disease because of the frequently asymptomatic nature and lack of sensitivity of diagnostic tests. Under conditions of immunosuppression (particularly systemic corticosteroid treatment), potentially fatal dissemination may occur. Thus, prevention and early diagnosis are important. Larva currens is a rare and pathognomonic cutaneous sign of strongyloidiasis, but is poorly described because of its unpredictable and fleeting occurrence. We report seven imported cases of larva currens seen in Paris between 1990 and 2020. We illustrate the clinical and biological features of this specific but uncommon sign of strongyloidiasis with clinical pictures. There were three males and four females, aged between 29 and 58 years. There were five migrants from endemic countries, one tourist and one expatriate. Digestive disorders were the main extracutaneous signs. All patients had eosinophilia above 0.5 G/L. All cases were confirmed by stool tests. All were cured with ivermectin. The rapidity and the short duration of the creeping eruption distinguish it from other parasitoses. Ivermectin is a treatment of choice. The key point is to think about preventing disseminated strongyloidiasis before giving corticosteroids not only among migrants but also among expatriates and tourists in endemic countries.

Parasitic larvae in bronchoalveolar lavage fluid in a non-immunocompromised patient.

Strongyloides stercoralis is responsible for a significant human parasitic infection known as strongyloidiasis. In addition, pulmonary strongyloidiasis is one of the most critical signs of disseminated strongyloidiasis. In this instance, S. stercoralis was unexpectedly discovered in bronchoalveolar lavage fluid.

Seroepidemiology of Strongyloides spp. Infection in Balimo, Western Province, Papua New Guinea.

Strongyloidiasis in Papua New Guinea (PNG) is poorly understood. There have been limited surveys describing the levels of endemicity in some regions of PNG, but in the Western Province, its occurrence and level of burden are unknown. This study aimed to determine the seroepidemiology of Strongyloides spp. seropositivity within a community located in the Balimo region of the Western Province. Plasma samples were collected from 120 adult participants and were subjected to anti-IgG Strongyloides spp. serological testing. Logistical regression analyses were performed to identify relationships between strongyloidiasis and attributes of sociodemography. In this cross-sectional cohort study, 22.5% (27/120; 95% CI: 15.9-30.8%) of participants were seropositive for strongyloidiasis. Participants with higher body mass indices were less likely to be seropositive for Strongyloides spp. infection (odds ratio [OR] = 0.85, P value = 0.008), and in the multivariable analysis, increasing units of age (adjusted OR [aOR] = 0.93, P value = 0.048) and participants ≤ 40 years old were associated with a decreased likelihood of Strongyloides spp. seropositivity (aOR = 0.07, P value = 0.034). The results from this study indicate that the occurrence of strongyloidiasis is high in the Western Province, PNG, and age is a determining factor of seroreactivity. This study provides evidence of endemic strongyloidiasis in this community and raises questions as to the impact of this neglected disease and other intestinal parasites on disease burden and comorbidities.

Prevalence of intestinal parasitic infections and genetic differentiation of Strongyloides stercoralis among migrant workers from Myanmar, Lao PDR and Cambodia in northeastern Thailand.

Intestinal parasitic infections (IPIs) remain a public-health problem worldwide, including in countries of the Lower Mekong subregion. Increases in human migration from neighboring countries might cause reemerging parasitic infections, leading to spread of parasites in the landscape. Here, we conducted a cross-sectional study to identify the prevalence of IPIs in migrant workers from Myanmar, Lao PDR, and Cambodia who were dwelling in Nakhon Ratchasima Province, northeastern Thailand. The identification of Strongyloides species and genetic differentiation of worms from migrant workers with different countries of origin was also assessed. Fresh stool samples were collected from 338 migrant workers and examined for evidence of IPIs using agar plate culture (APC) and the formalin-ethyl acetate concentration technique (FECT). Among those nine samples positive for nematodes by APC, the Strongyloides or hookworm species present was confirmed using the polymerase chain reaction (PCR) followed by DNA sequencing. This revealed eight cases of Strongyloides stercoralis infection and one of Necator americanus. Fifty-one out of 338 individuals (15.09%) were positive for IPIs using FECT and APC. Eggs of Opisthorchis-like flukes were the most common parasite (11.83% of samples), followed by S. stercoralis (2.37%), Entamoeba coli (1.50%), hookworm (0.89%), Taenia sp. (0.60%) and Hymenolepis nana (0.30%). The genetic differentiation of S. stercoralis recovered from migrant workers with different countries of origin was analyzed. Specimens of S. stercoralis isolated from workers from Lao PDR, Cambodia and Myanmar were genetically similar to those sequenced from Thailand. However, there were population-genetic differences between S. stercoralis from these Southeast Asian countries and other regions of the world. This study demonstrated that IPIs were prevalent in migrant workers in the northeastern region of Thailand. Our findings provided molecular confirmation of the presence of S. stercoralis and explored the genetic differentiation of S. stercoralis from those infected migrant workers. An effective anti-parasitic drug should be provided for migrant workers and its administration enforced.

Strongyloides stercoralis infestation mimicking recurrence of ovarian cancer in the liver.

Ovarian cancer is the most lethal gynecological cancer and can recur in most cases. Surgery is an option for recurrent ovarian cancer. Parasitic infestation disseminated in an immunocompromised host can be fatal. The case is here presented of a female patient diagnosed with early-stage ovarian cancer. Chemotherapy was initiated for treatment. At the follow-up examination, masses in the liver suggestive of recurrence were detected on positron emission tomography computed tomography. Surgery was performed. A Strongyloides stercoralis infestation mimicking relapsing ovarian cancer in the liver was diagnosed.

Clinical Performance of Real-Time Polymerase Chain Reaction for Strongyloides stercoralis Compared with Serology in a Nonendemic Setting.

Strongyloides stercoralis is a nematode endemic to subtropical and tropical regions that may cause asymptomatic carriage, peripheral eosinophilia, cutaneous, gastrointestinal, and pulmonary disease, or hyperinfection syndrome. Conventional diagnostic methods for strongyloidiasis include feces microscopy and culture, with low sensitivity in chronic infection due to the low helminth burden, and serology, which may be prone to false-negative results with immunocompromise and false-positive results with other infections and immunological disorders. We evaluated a laboratory-developed real-time polymerase chain reaction (RT-PCR), detecting the 18S SSU ribosomal RNA gene, compared with conventional diagnostic methods, using serology via ELISA as the gold-standard. The population studied included tertiary hospital inpatients and outpatients residing in a nonendemic area. Seven hundred fifty unfixed stool specimens submitted sequentially between 2014 and 2018 were tested for S. stercoralis via microscopy and RT-PCR. Agar plate culture (APC), Harada-Mori culture (HMC), and ELISA were performed in conjunction with 141, 135, and 177 of the specimens, respectively. RT-PCR yielded 13 positive and 730 negative results, with inhibition in seven specimens. ELISA yielded 53 positive, 18 equivocal, and 106 negative results. Results for direct diagnostic methods obtained after treatment with ivermectin were excluded from the performance analysis. Compared with ELISA, RT-PCR, microscopy, APC, and HMC exhibited sensitivities of 38%, 6%, 3%, and 0%, respectively, and specificities of 100%. Given the low sensitivities commensurate with testing a population with remote infection and thus low parasite burden, we recommend a combination of serological and molecular diagnostic testing to achieve the best balance of sensitivity and specificity.

Identification of miRNAs of Strongyloides stercoralis L1 and iL3 larvae isolated from human stool.

Strongyloidiasis is a neglected tropical disease caused by the soil-transmitted nematode by Strongyloides stercoralis, that affects approximately 600 million people worldwide. In immunosuppressed individuals disseminated strongyloidiasis can rapidly lead to fatal outcomes. There is no gold standard for diagnosing strongyloidiasis, and infections are frequently misdiagnosed. A better understanding of the molecular biology of this parasite can be useful for example for the discovery of potential new biomarkers. Interestingly, recent evidence showed the presence of small RNAs in Strongyloididae, but no data was provided for S. stercoralis. In this study, we present the first identification of miRNAs of both L1 and iL3 larval stages of S. stercoralis. For our purpose, the aims were: (i) to analyse the miRNome of L1 and iL3 S. stercoralis and to identify potential miRNAs of this nematode, (ii) to obtain the mRNAs profiles in these two larval stages and (iii) to predict potential miRNA target sites in mRNA sequences. Total RNA was isolated from L1 and iL3 collected from the stool of 5 infected individuals. For the miRNAs analysis, we used miRDeep2 software and a pipeline of bio-informatic tools to construct a catalog of a total of 385 sequences. Among these, 53% were common to S. ratti, 19% to S. papillosus, 1% to Caenorhabditis elegans and 44% were novel. Using a differential analysis between the larval stages, we observed 6 suggestive modulated miRNAs (STR-MIR-34A-3P, STR-MIR-8397-3P, STR-MIR-34B-3P and STR-MIR-34C-3P expressed more in iL3, and STR-MIR-7880H-5P and STR-MIR-7880M-5P expressed more in L1). Along with this analysis, we obtained also the mRNAs profiles in the same samples of larvae. Multiple testing found 81 statistically significant mRNAs of the total 1553 obtained (FDR < 0.05; 32 genes expressed more in L1 than iL3; 49 genes expressed more in L3 than iL1). Finally, we found 33 predicted mRNA targets of the modulated miRNAs, providing relevant data for a further validation to better understand the role of these small molecules in the larval stages and their valuein clinical diagnostics.

Using newly optimized genetic tools to probe Strongyloides sensory behaviors.

The oft-neglected human-parasitic threadworm, Strongyloides stercoralis, infects roughly eight percent of the global population, placing disproportionate medical and economic burden upon marginalized communities. While current chemotherapies treat strongyloidiasis, disease recrudescence and the looming threat of anthelminthic resistance necessitate novel strategies for nematode control. Throughout its life cycle, S. stercoralis relies upon sensory cues to aid in environmental navigation and coordinate developmental progression. Odorants, tastants, gases, and temperature have been shown to shape parasite behaviors that drive host seeking and infectivity; however, many of these sensory behaviors remain poorly understood, and their underlying molecular and neural mechanisms are largely uncharacterized. Disruption of sensory circuits essential to parasitism presents a promising strategy for future interventions. In this review, we describe our current understanding of sensory behaviors - namely olfactory, gustatory, gas sensing, and thermosensory behaviors - in Strongyloides spp. We also highlight the ever-growing cache of genetic tools optimized for use in Strongyloides that have facilitated these findings, including transgenesis, CRISPR/Cas9-mediated mutagenesis, RNAi, chemogenetic neuronal silencing, and the use of fluorescent biosensors to measure neuronal activity. Bolstered by these tools, we are poised to enter an era of rapid discovery in Strongyloides sensory neurobiology, which has the potential to shape pioneering advances in the prevention and treatment of strongyloidiasis.

The accuracy of a recombinant antigen immunochromatographic test for the detection of Strongyloides stercoralis infection in migrants from sub-Saharan Africa.

BACKGROUND: Strongyloidiasis, a nematode infection which is mainly caused by Strongyloides stercoralis in humans, can lead to a fatal syndrome in immunocompromised individuals. Its diagnosis is challenging due to the absence of a diagnostic gold standard. The infection is highly prevalent in migrants from endemic countries in tropical and subtropical areas, and a rapid diagnostic test would be helpful for screening purposes. The aim of this study was to estimate the accuracy of a novel immunochromatographic test (ICT) for the diagnosis of S. stercoralis infection. METHODS: A single-centre diagnostic accuracy study was undertaken using well-characterized frozen sera available from the biobank of a referral hospital for parasitic diseases in Italy. The included sera were from migrants from sub-Saharan Africa, and matching results were available for agar plate culture and/or polymerase chain reaction for S. stercoralis; moreover, the results of both a commercial enzyme-linked immunosorbent assay test and an in-house immunofluorescence test for strongyloidiasis were made available. Laboratory staff who read the ICT results were blinded as regards the results of the other tests. Two readers independently read the ICT, and a third one was involved when results were discrepant. The accuracy of the ICT was assessed both against the results of the panel of faecal tests and by latent class analysis (LCA). RESULTS: Agreement between the readers was excellent [Cohen's κ = 92.7%, 95% confidence interval (CI) 88.3-97.1%]. When assessed against the results of the faecal tests, the sensitivity and specificity of the ICT were 82.4% (95% CI 75.7-89.0%) and 73.8% (95% CI 66.8-80.9%), respectively. According to the LCA, the sensitivity and specificity were 86.3% (95% CI 80.1-92.5%) and 73.9% (95% CI 67.0-80.8%), respectively. CONCLUSIONS: The results of the ICT demonstrated ease of interpretation. The accuracy proved good, though the sensitivity might be further improved for screening purposes.