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Following almost 10 years of no reported cases, Guinea worm disease (GWD or dracunculiasis) reemerged in Chad in 2010 with peculiar epidemiological patterns and unprecedented prevalence of infection among non-human hosts, particularly domestic dogs. Since 2014, animal infections with Guinea worms have also been observed in the other three countries with endemic transmission (Ethiopia, Mali, and South Sudan), causing concern and generating interest in the parasites' true taxonomic identity and population genetics. We present the first extensive population genetic data for Guinea worm, investigating mitochondrial and microsatellite variation in adult female worms from both human and non-human hosts in the four endemic countries to elucidate the origins of Chad's current outbreak and possible host-specific differences between parasites. Genetic diversity of Chadian Guinea worms was considerably higher than that of the other three countries, even after controlling for sample size through rarefaction, and demographic analyses are consistent with a large, stable parasite population. Genealogical analyses eliminate the other three countries as possible sources of parasite reintroduction into Chad, and sequence divergence and distribution of genetic variation provide no evidence that parasites in human and non-human hosts are separate species or maintain isolated transmission cycles. Both among and within countries, geographic origin appears to have more influence on parasite population structure than host species. Guinea worm infection in non-human hosts has been occasionally reported throughout the history of the disease, particularly when elimination programs appear to be reaching their end goals. However, no previous reports have evaluated molecular support of the parasite species identity. Our data confirm that Guinea worms collected from non-human hosts in the remaining endemic countries of Africa are Dracunculus medinensis and that the same population of worms infects both humans and dogs in Chad. Our genetic data and the epidemiological evidence suggest that transmission in the Chadian context is currently being maintained by canine hosts.

The prevalence and diversity of parasitic nematodes in wildlife have been well studied for certain species, yet for others considerable gaps in knowledge exist. The parasitic nematode Dracunculus insignis infects North American wildlife, and past research on this species has led to an increased understanding of the potential host diversity and transmission of the closely related human Guinea worm, Dracunculus medinensis (which is currently the focus of a global eradication program). Many definitive hosts have been documented for D. insignis; however, the life cycle has been studied only in laboratories, and only a single phylogenetic study has been conducted on D. insignis (from Canada). The goals of the present study were to investigate the prevalence of infections with Dracunculus species among wildlife at a single site (Di-Lane plantation) in the southeastern United States, evaluate the genetic diversity of parasites at this site, and investigate potential paratenic hosts that may be involved in transmission. Over 3 yr, we sampled 228 meso-mammals, reporting an overall prevalence of infection with Dracunculus insignis of 20% (46/228). Amphibians and fish were sampled in the same geographic area as infected meso-mammals. Dracunculus insignis third-stage larvae were recovered from 2 different species of amphibians, but all fish sampled were negative. Phylogenetic analysis of the partial cytochrome c oxidase I (COI) gene showed very little diversity of Dracunculus at Di-Lane; however, we did recover a single nematode from a Virginia opossum (Didelphis virginiana) that falls outside of the D. insignis clade, more closely aligns with Dracunculus lutrae, and may represent an undescribed species. This work documents the occurrence of D. insignis in frogs, a potential transmission pathway for D. insignis at a single geographic site in nature. When applied to the global Guinea Worm Eradication Program, and Chad, Africa, in particular, this work increases our knowledge of the potential role of aquatic animals in the transmission of Dracunculus species and informs on potential intervention strategies that may be applied to the eradication of Guinea worm in Africa.

Background: Guinea worm-Dracunculus medinensis-was historically one of the major parasites of humans and has been known since antiquity. Now, Guinea worm is on the brink of eradication, as efforts to interrupt transmission have reduced the annual burden of disease from millions of infections per year in the 1980s to only 54 human cases reported globally in 2019. Despite the enormous success of eradication efforts to date, one complication has arisen. Over the last few years, hundreds of dogs have been found infected with this previously apparently anthroponotic parasite, almost all in Chad. Moreover, the relative numbers of infections in humans and dogs suggests that dogs are currently the principal reservoir on infection and key to maintaining transmission in that country.Principal findings: In an effort to shed light on this peculiar epidemiology of Guinea worm in Chad, we have sequenced and compared the genomes of worms from dog, human and other animal infections. Confirming previous work with other molecular markers, we show that all of these worms are D. medinensis, and that the same population of worms are causing both infections, can confirm the suspected transmission between host species and detect signs of a population bottleneck due to the eradication efforts. The diversity of worms in Chad appears to exclude the possibility that there were no, or very few, worms present in the country during a 10-year absence of reported cases.Conclusions: This work reinforces the importance of adequate surveillance of both human and dog populations in the Guinea worm eradication campaign and suggests that control programs aiming to interrupt disease transmission should stay aware of the possible emergence of unusual epidemiology as pathogens approach elimination.

After a ten-year absence of reported Guinea worm disease in Chad, human cases were rediscovered in 2010, and canine cases were first recorded in 2012. In response, active surveillance for Guinea worm in both humans and animals was re-initiated in 2012. As of 2018, the Chad Guinea Worm Eradication Program (CGWEP) maintains an extensive surveillance system that operates in 1,895 villages, and collects information about worms, hosts (animals and humans), and animal owners. This report describes in detail the CGWEP surveillance system and explores epidemiological trends in canine Guinea worm cases during 2015-2018. Our results showed an increased in the number of canine cases detected by the system during the period of interest. The proportion of worms that were contained (i.e., water contamination was prevented) improved significantly over time, from 72.8% in 2015 to 85.7% in 2018 (Mantel-Haenszel chi-square = 253.3, P < 0.0001). Additionally, approximately 5% of owners of infected dogs reported that the dog had a Guinea worm-like infection earlier that year; 12.6% had a similar worm in a previous year. The proportion of dogs with a history of infection in a previous year increased over time (Mantel-Haenszel chi-square = 18.8, P < 0.0001). Canine cases were clustered in space and time: most infected dogs (80%) were from the Chari Baguirmi (38.1%) and Moyen Chari Regions (41.9%), and for each year the peak month of identified canine cases was June, with 78.5% occurring during March through August. Findings from this report evoke additional questions about why some dogs are repeatedly infected. Our results may help to target interventions and surveillance efforts in terms of space, time, and dogs susceptible to recurrent infection, with the ultimate goal of Guinea worm eradication.

Adult female guinea worms (Dracunculus spp.) usually are reported to occur singly in the extremities of their hosts, from which they deliver their larvae into the water through fistulae in the host's skin. We visually examined for the presence of worms in the fascia of the limbs of skinned carcasses of 184 river otters (Lontra canadensis) harvested in Arkansas and report observations of cysts on wrists and ankles found on 12 otters. Cysts averaged 15.6 × 24.6 mm in diameter, were round to oval, and contained masses of up to 19 adult female Dracunculus sp. (mean 6.7). We speculate that high levels of infection in consumed paratenic hosts might have caused high infection rates, leading to large cyst formation in otters, as larvigerous Dracunculus sp. females accumulated in extremities. No males were discovered during the study, so identification of a sample of worms was based on molecular techniques.

Transmission of dracunculiasis (Guinea worm disease), a waterborne, parasitic disease targeted for eradication, was thought to have been interrupted in Chad since 2000, when the last case was reported. However, in 2010, 10 cases were confirmed by the Chad Ministry of Public Health (Ministère de la Santé Publique [MSP]) and the World Health Organization (WHO) during field investigations in which rumored cases were investigated and nearby villages were actively searched for additional cases. Because patients were not prevented from contaminating water sources, new cases were expected in 2011. During January-February 2011, MSP, WHO, and CDC conducted an investigation to gather additional information to guide prevention and response activities before the 2011 transmission season. Seven districts where cases had been confirmed or suspected in 2010 or where dracunculiasis was endemic during 1994-2000 were surveyed. The results of those surveys indicated that residents of 116 (55%) of 210 villages and 13 (87%) of 15 nomad camps consumed water from unsafe sources; 157 (75%) of 209 village key informants (KIs) and five (33%) of 15 nomad camp KIs knew about dracunculiasis. Thirty-one villages had confirmed or suspected cases during 2009-2011 and were classified as at-risk, requiring weekly active surveillance and urgent pre-positioning of materials for the 2011 transmission season. Nomadic populations are at risk for dracunculiasis because of unsafe water consumption and minimal knowledge of the disease. These populations also require targeted surveillance and prevention efforts (e.g., filter distribution, education, and case containment) to interrupt dracunculiasis transmission .

Plant parasitic cyst nematodes induce specific hypermetabolic syncytial nurse cell structures in host roots. A characteristic feature of syncytia is the lack of the central vacuole and the formation of numerous small and larger vesicles. We show that these structures are formed de novo via widening of ER cisternae during the entire development of syncytium, whereas in advanced stages of syncytium development, larger vacuoles are also formed via fusion of vesicles/tubules surrounding organelle-free pre-vacuole regions. Immunogold transmission electron microscopy of syncytia localised the vacuolar markers E subunit of vacuolar H+-adenosinetriphosphatase (V-ATPase) complex and tonoplast intrinsic protein (γ-TIP1;1) mostly in membranes surrounding syncytial vesicles, thus indicating that these structures are vacuoles and that some of them have a lytic character. To study the function of syncytial vacuoles, changes in expression of AtVHA-B1, AtVHA-B2 and AtVHA-B3 (coding for isoforms of subunit B of V-ATPase), and TIP1;1 and TIP1;2 (coding for γ-TIP proteins) genes were analysed. RT-qPCR revealed significant downregulation of AtVHA-B2, TIP1;1 and TIP1;2 at the examined stages of syncytium development compared to uninfected roots. Expression of VHA-B1 and VHA-B3 decreased at 3 dpi but reached the level of control at 7 dpi. These results were confirmed for TIP1;1 by monitoring At-γ-TIP-YFP reporter construct expression. Infection test conducted on tip1;1 mutant plants showed formation of larger syncytia and higher numbers of females in comparison to wild-type plants indicating that reduced levels or lack of TIP1;1 protein promote nematode development.

Background Wolbachia endosymbionts are a proven target for control of human disease caused by filarial nematodes. However, little is known about the occurrence of Wolbachia in taxa closely related to the superfamily Filarioidea. Our study addressed the status of Wolbachia presence in members of the superfamily Dracunculoidea by screening the human parasite Dracunculus medinensis and related species from wildlife for Wolbachia . Findings D. medinensis , D. lutrae and D. insignis specimens were all negative for Wolbachia colonization by PCR screening for the Wolbachia ftsZ , 16S rRNA and Wolbachia surface protein ( wsp ) sequences. The quality and purity of the DNA preparations was confirmed by amplification of nematode 18S rRNA and cytochrome c oxidase subunit I sequences. Furthermore, Wolbachia endobacteria were not detected by whole mount fluorescence staining, or by immunohistochemistry using a Wolbachia -specific antiserum. In contrast, positive control Brugia malayi worms were shown to harbour Wolbachia by PCR, fluorescence staining and immunohistochemistry. Conclusions Three examined species of Dracunculus showed no evidence of Wolbachia endobacteria. This supports that members of the superfamily Dracunculoidea are free of Wolbachia . Within the order Spirurida, these endosymbionts appear restricted to the Filarioidea.

In this study, in vitro drug testing was combined with proteomic and bioinformatic analyses to identify and characterize proteins involved in larval development of Oesophagostomum dentatum, an economically important parasitic nematode. Four hydrolase inhibitors ο-phenanthroline, sodium fluoride, iodoacetamide and 1,2-epoxy-3-(pnitrophenoxy)-propane (EPNP) significantly inhibited (≥90%) larval development. Comparison of the proteomic profiles of the development-inhibited larvae with those of uninhibited control larvae using two-dimensional gel electrophoresis, and subsequent MALDI-TOF mass spectrometric analysis identified a down-regulation of 12 proteins inferred to be involved in various larval developmental processes, including post-embryonic development and growth. Furthermore, three proteins (i.e. intermediate filament protein B, tropomyosin and peptidyl-prolyl cis-trans isomerase) inferred to be involved in the moulting process were down-regulated in moulting- and development-inhibited O. dentatum larvae. This first proteomic map of O. dentatum larvae provides insights in the protein profile of larval development in this parasitic nematode, and significantly improves our understanding of the fundamental biology of its development. The results and the approach used might assist in developing new interventions against parasitic nematodes by blocking or disrupting their key biological pathways.