Strongyloides stercoralis differs from the other soil-transmitted helminths because it puts infected subjects at risk of a fatal syndrome (in cases of immunosuppression for medical conditions, immunosuppressant therapies, or both). Chronic strongyloidiasis is often a non-severe condition, or is sometimes even asymptomatic, but diagnosis and effective therapy are essential in order to eradicate the infection and the life-long risk involved. Therefore, diagnostic methods need to be highly sensitive. Stool microscopy and the Kato-Katz technique are commonly used in prevalence studies, but they are inadequate for S. stercoralis detection. This is probably the main reason why the global prevalence has long been underestimated. Concentration methods, the Baermann technique and Koga agar plate culture have better, but still unsatisfactory, sensitivity. Serological tests have demonstrated higher sensitivity; although some authors have concerns about their specificity, it is possible to define cut-off values over which infection is almost certain. In particular, the luciferase immunoprecipitation system technique combined with a recombinant antigen (NIE) demonstrated a specificity of almost 100%. ELISA coproantigen detection has also shown promising results, but still needs full evaluation. Molecular diagnostic methods are currently available in a few referral centres as in-house techniques. In this review, on the basis of the performance of the different diagnostic methods, we outline diagnostic strategies that could be proposed for different purposes, such as: prevalence studies in endemic areas; individual diagnosis and screening; and monitoring of cure in clinical care and clinical trials.

2-Oxoglutarate (2OG) and Fe(II)-dependent oxygenase domain-containing protein 1 (OGFOD1) is predicted to be a conserved 2OG oxygenase, the catalytic domain of which is related to hypoxia-inducible factor prolyl hydroxylases. OGFOD1 homologs in yeast are implicated in diverse cellular functions ranging from oxygen-dependent regulation of sterol response genes (Ofd1, Schizosaccharomyces pombe) to translation termination/mRNA polyadenylation (Tpa1p, Saccharomyces cerevisiae). However, neither the biochemical activity of OGFOD1 nor the identity of its substrate has been defined. Here we show that OGFOD1 is a prolyl hydroxylase that catalyzes the posttranslational hydroxylation of a highly conserved residue (Pro-62) in the small ribosomal protein S23 (RPS23). Unusually OGFOD1 retained a high affinity for, and forms a stable complex with, the hydroxylated RPS23 substrate. Knockdown or inactivation of OGFOD1 caused a cell type-dependent induction of stress granules, translational arrest, and growth impairment in a manner complemented by wild-type but not inactive OGFOD1. The work identifies a human prolyl hydroxylase with a role in translational regulation. Significance Members of the 2-oxoglutarate (2OG)-dependent oxygenase superfamily catalyze a range of important biological oxidations. Structurally informed bioinformatic predictions suggest that the human genome encodes as yet unassigned members of the superfamily. We describe work demonstrating that 2OG and Fe(II)-dependent oxygenase domain-containing protein 1 (OGFOD1) is a protein hydroxylase that modifies the small ribosomal subunit protein RPS23 at a conserved prolyl residue in the ribosome-decoding center and that suppression or deletion of OGFOD1 is associated with the activation of translational stress pathways. Together with studies of homologous genes in flies and yeast described in accompanying manuscripts, the work identifies a unique function for 2OG oxygenase-catalyzed hydroxylation in ribosome biology.

Schistosomiasis is a neglected infection affecting millions of people, mostly living in sub-Saharan Africa. Morbidity and mortality due to chronic infection are relevant, although schistosomiasis is often clinically silent. Different diagnostic tests have been implemented in order to improve screening and diagnosis, that traditionally rely on parasitological tests with low sensitivity. Aim of this study was to evaluate the accuracy of different tests for the screening of schistosomiasis in African migrants, in a non endemic setting. A retrospective study was conducted on 373 patients screened at the Centre for Tropical Diseases (CTD) in Negrar, Verona, Italy. Biological samples were tested with: stool/urine microscopy, Circulating Cathodic Antigen (CCA) dipstick test, ELISA, Western blot, immune-chromatographic test (ICT). Test accuracy and predictive values of the immunological tests were assessed primarily on the basis of the results of microscopy (primary reference standard): ICT and WB resulted the test with highest sensitivity (94% and 92%, respectively), with a high NPV (98%). CCA showed the highest specificity (93%), but low sensitivity (48%). The analysis was conducted also using a composite reference standard, CRS (patients classified as infected in case of positive microscopy and/or at least 2 concordant positive immunological tests) and Latent Class Analysis (LCA). The latter two models demonstrated excellent agreement (Cohen’s kappa: 0.92) for the classification of the results. In fact, they both confirmed ICT as the test with the highest sensitivity (96%) and NPV (97%), moreover PPV was reasonably good (78% and 72% according to CRS and LCA, respectively). ELISA resulted the most specific immunological test (over 99%). The ICT appears to be a suitable screening test, even when used alone. The rapid test ICT was the most sensitive test, with the potential of being used as a single screening test for African migrants. Schistosomiasis is probably the most important of the neglected tropical diseases (NTD) caused by helminthes (worms). It is acquired bathing in freshwater in endemic areas. The life cycle is complex and involves freshwater snails. Schistosomiasis, caused by Schistosoma mansoni, S. haematobium and less frequently by other species, affects more than 200 million people, mostly in Africa, and may chronically cause irreversible damage of the liver (S. mansoni) or of the kidneys and the urinary tract, including cancer of the bladder (S. haematobium). As in chronic infections eggs of both species are often missed by microscopy of faeces and urine, with this retrospective study we evaluate the accuracy of different, alternative diagnostic tests, for the screening of schistosomiasis in African migrants and asylum seekers, of whom many thousands reach the Italian coast every year proceding from the most endemic areas. The most interesting finding of our study is that a rapid diagnostic test for antibody detection in blood, easy to use as a point-of-care tool, resulted the most sensitive of the five tests evaluated, and thus is very promising as a screening tool even when used without any additional test.

Strongyloidiasis is a neglected tropical disease caused by the human infective nematodes Strongyloides stercoralis, Strongyloides fuelleborni fuelleborni and Strongyloides fuelleborni kellyi. Previous large-scale studies exploring the genetic diversity of this important genus have focused on Southeast Asia, with a small number of isolates from the USA, Switzerland, Australia and several African countries having been genotyped. Consequently, little is known about the global distribution of geographic sub-variants of these nematodes and the genetic diversity that exists within the genus Strongyloides generally. We extracted DNA from human, dog and primate feces containing Strongyloides, collected from several countries representing all inhabited continents. Using a genotyping assay adapted for deep amplicon sequencing on the Illumina MiSeq platform, we sequenced the hyper-variable I and hyper-variable IV regions of the Strongyloides 18S rRNA gene and a fragment of the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene from these specimens. We report several novel findings including unique S. stercoralis and S. fuelleborni genotypes, and the first identifications of a previously unknown S. fuelleborni infecting humans within Australia. We expand on an existing Strongyloides genotyping scheme to accommodate S. fuelleborni and these novel genotypes. In doing so, we compare our data to all 18S and cox1 sequences of S. fuelleborni and S. stercoralis available in GenBank (to our knowledge), that overlap with the sequences generated using our approach. As this analysis represents more than 1,000 sequences collected from diverse hosts and locations, representing all inhabited continents, it allows a truly global understanding of the population genetic structure of the Strongyloides species infecting humans, non-human primates, and domestic dogs. Strongyloidiasis is a neglected tropical disease caused by the human infective worms (nematodes) Strongyloides stercoralis, Strongyloides fuelleborni fuelleborni and Strongyloides fuelleborni kellyi. Little is known about the genetic diversity of these nematodes and the possibility of geographically isolated genetic types is a particularly interesting research question, given the unique life cycle of these worms which includes both sexual and asexual stages. We extracted DNA from human, dog and primate feces containing Strongyloides, collected from several countries representing all inhabited continents. Using next-generation sequencing, we analyzed three Strongyloides DNA fragments from these specimens; two fragments of the 18S gene and one from the cytochrome c oxidase subunit 1 (cox1) gene. Using this approach, we discovered some unique S. stercoralis and S. fuelleborni genotypes, and identified a previously unknown S. fuelleborni infecting humans within Australia. We compared our data to all 18S and cox1 sequences of S. fuelleborni and S. stercoralis available in public databases and identified several patterns relating to the global distribution of certain genotypes. This knowledge could allow us to infer the origin of human Strongyloides infections in the future, and assess the role certain animals (non-human primates and dogs) might play in the transmission of Strongyloides to humans.

Data from recent studies support the hypothesis that infections by human gastrointestinal (GI) helminths impact, directly and/or indirectly, on the composition of the host gut microbial flora. However, to the best of our knowledge, these studies have been conducted in helminth-endemic areas with multi-helminth infections and/or in volunteers with underlying gut disorders. Therefore, in this study, we explore the impact of natural mono-infections by the human parasite Strongyloides stercoralis on the faecal microbiota and metabolic profiles of a cohort of human volunteers from a non-endemic area of northern Italy (S+), pre- and post-anthelmintic treatment, and compare the findings with data obtained from a cohort of uninfected controls from the same geographical area (S−). Analyses of bacterial 16S rRNA high-throughput sequencing data revealed increased microbial alpha diversity and decreased beta diversity in the faecal microbial profiles of S+ subjects compared to S−. Furthermore, significant differences in the abundance of several bacterial taxa were observed between samples from S+ and S− subjects, and between S+ samples collected pre- and post-anthelmintic treatment. Faecal metabolite analysis detected marked increases in the abundance of selected amino acids in S+ subjects, and of short chain fatty acids in S− subjects. Overall, our work adds valuable knowledge to current understanding of parasite-microbiota associations and will assist future mechanistic studies aimed to unravel the causality of these relationships.

SUMMARY: Strongyloides stercoralisis a parasite that can cause death in immunocompromised people. A proper diagnosis is hence essential. The real-time polymerase-chain reaction (RT–PCR) is a novel, promising diagnostic method, that detects the DNA of the parasite in stool samples. In this retrospective study, we compared the sensitivity of agar plate coproculture (APC), an in-house immunofluorescence test (IFAT) and an in-house RT–PCR for the diagnosis ofS. stercoralisinfection. The study sample was composed by 223 samples. Samples resulting positive to APC, IFAT and RT–PCR were 20, 140 and 25, respectively. When sensitivity was calculated against a composite reference standard, serology confirmed the best performance (sensitivity 95%), followed by RT–PCR (57%) and APC (45%). In conclusion, in a non-endemic setting, serology is the best screening method, while the combination of APC and RT–PCR does not seem a reasonable approach to increase sensitivity. Both methods can have a role as confirmatory tests for selected cases.