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.

Four species of Strongyloides, Strongyloides felis, Strongyloides planiceps, Strongyloides stercoralis and Strongyloides tumefaciens, have been identified in cats based on morphology and location in the host with limited data on the prevalence and disease potential of these different species. Strongyloides tumefaciens adults are located in colonic nodules while the other three species are in the small intestine. The literature on Strongyloides in cats is scattered and has never been compiled. The aim of this article is to provide a short review of the existing literature on Strongyloides spp. in cats, to describe the pathology of colonic nodules containing Strongyloides sp. seen at necropsies of cats in St. Kitts, West Indies, and to provide the first unequivocal report of zoonotic S. stercoralis in cats based on sequencing analysis of a portion of the cytochrome c oxidase subunit 1 (cox1) gene, and supported by phylogenetic analysis. Colonic nodules containing sections of nematodes, histologically compatible with Strongyloides sp. were seen during necropsy in six cats in St. Kitts, West Indies. Sequencing of the cox1 gene of the mitochondrial DNA extracted from colonic nodules from two of these cats matched sequences of the zoonotic strain of S. stercoralis. The morphological similarities between S. stercoralis-associated colonic nodules and previous reports of S. tumefaciens, together with the insufficient defining criteria for S. tumefaciens raises questions about the validity of the species. Further sampling and genetic characterization of isolates is needed to understand the species in cats and their zoonotic potential.

Background: Current recommendations for diagnosis of Dirofilaria immitis infection in dogs rely on the detection of antigen produced largely by adult females coupled with the visualization of microfilariae (mf) in the circulation via a microfilaria detection test (MFDT). It is hypothesized that qPCR assays used in parallel with antigen detection tests will perform better in detecting mf than modified Knott’s test (MK), when combined with antigen detection. This study compares probe-based qPCR and MK techniques for mf detection used in parallel with the DiroCHEK^® antigen test to screen for heartworm infection in shelter dogs. Methods: Matching blood and serum samples were collected from 300 shelter dogs in Brazos and Harris counties, Texas, USA. Blood was assessed for the presence of mf via MK and the presence of D. immitis DNA by a species-specific probe-based qPCR assay. Serum samples were tested for the presence of heartworm antigen using DiroCHEK^® before and after immune complex dissociation (ICD) via heat treatment. In addition, the performance of each diagnostic test was evaluated via Chi-square test, Cochran’s Q test, and post hoc analysis. Results: Qualitatively, MK detected mf in 22.0% (66/300) of samples, 55 of which were morphologically identified as D. immitis and 11 as Acanthocheilonema reconditum. The range of heartworm mf was 28 to 88,803 mf/ml (median: 6627.5). Real-time PCR detected D. immitis DNA in 20.7% (62/300) of samples. Heartworm antigen was detected in 24.7% (74/300) of samples pre-ICD, and in 29.3% (88/300) post-ICD. When comparing tests, the Chi-square and McNemar’s tests showed that the difference between positive and negative proportions was statistically significant. The Cochran test showed the difference in the distributions of cases and non-cases was significant when individual tests were combined (χ² = 62.3, df = 3, P < 0.0001) and when parallel methods were combined (χ² = 43.1, df = 4, P < 0.0001). Conclusion: Considering individual and combined test performances, practicality, and efficient use of bench time, this heartworm-specific probe-based qPCR method is a viable option as a mf detection test to be used in parallel with antigen tests for canine heartworm infection in diagnostic and research settings. Graphical

The Onchocerca lupi nematode is an emerging helminth capable of infecting pets and humans. We detected this parasite in 2 dogs that were imported into Canada from the southwestern United States, a region to which this nematode is endemic. We discuss risk for establishment of O. lupi in Canada.