Potential Impact of a Diagnostic Test for Detecting Prepatent Guinea Worm Infections in Dogs.

Chad has seen a considerable reduction in cases of Guinea worm disease (or dracunculiasis) in domestic dogs in recent years. Tethering of dogs and application of Abate® larvicide to water sources appear to have contributed to this progress, but with 767 reported dog cases in 2021, accelerating elimination of the disease in Chad may require additional tools. We investigate the potential benefits of a hypothetical diagnostic test that could be capable of detecting prepatent infections in dogs. We adapt an agent-based simulation model for forecasting the impact of interventions on guinea worm disease in dogs to examine the interaction of multiple test factors including test accuracy, when the test can detect infection, dog selection, and dog-owner compliance with tethering recommendations. We find that a diagnostic test could be successful if used in conjunction with existing interventions, and elimination can be achieved within 2 years with 80% or higher test sensitivity, 90% or higher specificity, systematic testing of each dog twice per year, and more than 90% long-term tethering compliance when a dog tests positive or a worm is emerging. Because of the long incubation period of Guinea worm disease (10-14 months) and the fact that no treatment exists, the benefits of the test rely on the testing rollout and response of dog owners. If the test could estimate the timing of worm emergence, long-term tethering could be eliminated and infected dogs could be tethered only when the worms are expected, minimizing the related resources (human and financial) to support the intervention.

Unusual cutaneous manifestations of dracunculiasis: Two rare case reports.

A nematode parasite, Dracunculus medinensis, causes dracunculiasis. Despite being non-fatal, this condition causes significant morbidity. Dracunculiasis is considered an eradicated disease in India since 1999. We report two cases that document the unusual linear morphea-like morphology of the calcified D. medinensis and the rare periorbital location of the worm. The cases presented here are rare and a diagnostic challenge, considering the eradicated status of dracunculiasis.

Development and validation of a quantitative PCR for the detection of Guinea worm (Dracunculus medinensis).

Dracunculus medinensis (Guinea worm) is a parasitic nematode that can cause the debilitating disease dracunculiasis (Guinea worm disease) in humans. The global Guinea Worm Eradication Program has led intervention and eradication efforts since the 1980s, and Guinea worm infections in people have decreased >99.99%. With the final goal of eradication drawing nearer, reports of animal infections from some remaining endemic countries pose unique challenges. Currently, confirmation of suspected Guinea worm infection relies on conventional molecular techniques such as polymerase chain reaction (PCR), which is not specific to Guinea worm and, therefore, requires sequencing of the PCR products to confirm the identity of suspect samples, a process that often takes a few weeks. To decrease the time required for species confirmation, we developed a quantitative PCR assay targeting the mitochondrial cytochrome b (cytb) gene of Guinea worm. Our assay has a limit of detection of 10 copies per reaction. The mean analytical parameters (± SE) were as follows: efficiency = 93.4 ± 7.7%, y-intercept = 40.93 ± 1.11, slope = -3.4896 ± 0.12, and the R2 = 0.999 ± 0.004. The assay did not amplify other nematodes found in Guinea worm-endemic regions and demonstrated 100% diagnostic sensitivity and specificity. Implementation of this quantitative PCR assay for Guinea worm identification could eliminate the need for DNA sequencing to confirm species. Thus, this approach can be implemented to provide more rapid confirmation of Guinea worm infections, leading to faster execution of Guinea worm interventions while increasing our understanding of infection patterns.

Human infection with an unknown species of Dracunculus in Vietnam.

Guinea worm (GW) disease, caused by Dracunculus medinensis, is an almost eradicated waterborne zoonotic disease. The World Health Organization (WHO) currently lists GW as endemic in only five African countries. In July 2020, the Vietnamese public health surveillance system detected a hanging worm in a 23-year-old male patient, who did not report any travel to Africa or any country previously endemic for GW. The patient was hospitalized with symptoms of fatigue, anorexia, muscle aches, and abscesses, with worms hanging out of the skin in the lower limbs. The worms were retrieved from the lesions and microscopically examined in Vietnam, identifying structures compatible with Dracunculus spp. and L1-type larvae. A section of this parasite was sent to the Centers for Disease Control and Prevention (CDC) in Atlanta, United States, for confirmatory diagnosis of GW. The adult worm had cuticle structures compatible with Dracunculus parasites, although the length of L1 larvae was about 339 µm, substantially shorter than D. medinensis. DNA sequence analysis of the 18S small subunit rRNA gene confirmed that this parasite was not GW, and determined that the sample belonged to a Dracunculus sp. not previously reported in GenBank that clustered with the animal-infective Dracunculus insignis and Dracunculus lutrae, located in a different clade than D. medinensis. This study highlights the importance of effective public health surveillance systems and the collaborative work of local public health authorities from Vietnam with the WHO and CDC in efforts to achieve the eradication of GW.

Development of a Multiplex Bead Assay for the Detection of Canine IgG4 Antibody Responses to Guinea Worm.

Increased levels of guinea worm (GW) disease transmission among dogs in villages along the Chari River in Chad threaten the gains made by the GW Eradication Program. Infected dogs with preemergent worm blisters are difficult to proactively identify. If these dogs are not contained, blisters can burst upon submersion in water, leading to the contamination of the water supply with L1 larvae. Guinea worm antigens previously identified using sera from human dracunculiasis patients were coupled to polystyrene beads for multiplex bead assay analysis of 41 non-endemic (presumed negative) dog sera and 39 sera from GW-positive dogs from Chad. Because commercially available anti-dog IgG secondary antibodies did not perform well in the multiplex assay, dog IgGs were partially purified, and a new anti-dog IgG monoclonal antibody was developed. Using the new 4E3D9 monoclonal secondary antibody, the thioredoxin-like protein 1-glutathione-S-transferase (GST), heat shock protein (HSP1)-GST, and HSP2-GST antigen multiplex assays had sensitivities of 69-74% and specificities of 73-83%. The domain of unknown function protein 148 (DUF148)-GST antigen multiplex assay had a sensitivity of 89.7% and a specificity of 85.4%. When testing samples collected within 1 year of GW emergence (n = 20), the DUF148-GST assay had a sensitivity of 90.0% and a specificity of 97.6% with a receiver-operating characteristic area under the curve of 0.94. Using sera from two experimentally infected dogs, antibodies to GW antigens were detected within 6 months of exposure. Our results suggest that, when used to analyze paired, longitudinal samples collected 1-2 months apart, the DUF148/GST multiplex assay could identify infected dogs 4-8 months before GW emergence.

Development of a Multiplex Bead Assay for the Detection of IgG Antibody Responses to Guinea Worm.

The success of the Guinea Worm (GW) Eradication Program over the past three decades has been tempered by the persistence of GW disease in a few African nations and the potential for a future resurgence in cases. Domestic dogs are now a major concern as a disease reservoir as large numbers of cases of canine GW disease are now reported each year, mainly along the Chari River in Chad. As a first step toward the development of a serologic assay for dogs, archived human plasma samples from dracunculiasis-positive donors from Togo were used to select adult female GW antigens for peptide sequencing and cloning. Eight protein sequences of interest were expressed as recombinant glutathione-S-transferase (GST) fusion proteins, and the most promising proteins were coupled to carboxylated microspheres for use in multiplex assays. A thioredoxin-like protein (TRXL1) and a domain of unknown function (DUF148) were assessed for total IgG and IgG4 reactivities using a panel of specimens from GW cases, uninfected donors, and individuals infected with various nematode worms, including Onchocerca volvulus. Both the DUF148-GST and the TRXL1-GST assays cross-reacted with O. volvulus sera, but the latter assay was always the more specific. The IgG4 and total IgG TRXL1-GST assays both had sensitivities > 87% and specificities > 90%. Maximum specificity (> 96%) was obtained with the total IgG assay when reactivity to both antigens was used to define a positive case. Given the good performance of the human assay, we are now working to modify the assay for dog assessments.

Guinea-worm (Dracunculus medinensis) infection presenting as a diabetic foot abscess: A case report from Kerala.

Dracunculiasis or guinea-worm infection is a water-borne, parasitic disease that can cause major morbidity. Dracunculiasis in patients with diabetes can be misdiagnosed as a diabetic foot abscess, which is a common complication of poorly controlled diabetes. This is a report of guinea-worm disease (GWD) in a 57-year-old man with diabetes from a rural area of Kerala. There is need for awareness among physicians about the occurrence of GWD in people with diabetes and the need to ensure supply of safe drinking water to prevent its re- emergence. Though WHO has declared India free of GWD, a few cases have been reported from the country.

First report of Dracunculus insignis in two naturally infected cats from the northeastern USA.

Dracunculiasis is rarely reported in cats, yet over the last few years we have identified two cats with filarioid-like spirurid infections. Case 1 was a 9-year-old cat with pituitary-dependent hyperadrenocorticism from New York tate from which four adult dracunculoid nematodes were isolated from its torso. Based on morphometric characteristics and parasite geographic distribution, the specimens were identified as Dracunculus insignis females; at least one of the females was gravid, suggestive of patent infection. Species identification was confirmed through amplification and sequence analysis of nuclear and mitochondrial loci. Case 2 was a 14-year-old diabetic cat from Massachusetts. Formalin-fixed sections were obtained from a subcutaneous mass excised from the left foreleg. Histopathological examination revealed a large nematode with morphometrical characteristics of Dracunculus, surrounded by lymphocytes and sheets of eosinophils. These two cases appear to be the first published reports of dracunculiasis in domestic cats in the USA, and based on the findings from case 1, D insignis may be the species associated with both infections.

Parasitic infestations requiring surgical interventions.

Parasitic infestation is common in developing countries especially in Africa. Children are often more vulnerable to these infections. Many health problems result from these infestations, including malnutrition, iron-deficiency anemia, surgical morbidities, and even impaired cognitive function and educational achievement. Surgical intervention may be needed to treat serious complications caused by some of these parasites. Amoebic colitis and liver abscess caused by protozoan infections; intestinal obstruction, biliary infestation with cholangitis and liver abscess, and pancreatitis caused by Ascaris lumbricoides; biliary obstruction caused by Faschiola; hepatic and pulmonary hydatid cysts caused by Echinococcus granulosus and multilocularis are examples. Expenditure of medical care of affected children may cause a great burden on many African governments, which are already suffering from economic instability. The clinical presentation, investigation, and management of some parasitic infestations of surgical relevance in African children are discussed in this article.

Emergence of Onchocerca volvulus from skin mimicking Dracunculiasis medinensis.

We describe 11 cases of suspected Dracunculus medinensis infection in which the worm recovered was identified as Onchocerca volvulus. Identification was based on morphology of the examined specimen.

[Imported dracunculiasis: four cases confirmed in the south of Algeria]. / La dracunculose d'importation: quatre cas confirmés dans le sud algérien.

The authors report for the first time, in the south of Algeria, four imported cases of dracunculiasis in Malian Touaregs. The patients came from Mali. They remained two months in Tamanrasset before settling in Illizi where they have been working illegally for more than nine months. In August 2007, because of infected lesions appearing on their legs a month earlier, the patients decided to go to the Illizi hospital for treatment The diagnosis of dracunculiasis was made by direct observation of worms emerging from lesions and by microscopic examination of larvae. The patients were then isolated and treated. The Ministry of Health immediately took measures to avoid local transmission of the disease, never seen in the region. The measures consisted in increasing awareness campaign among medical and paramedical staff about dracunculiasis and in pointing out the importance of detection of new cases among nomadic populations. The risk of emergence of a new dracunculiasis indigenous focus in the south of Algeria should be seriously considered, especially as the number of immigrants from endemic countries is increasing considerably

Dracunculiasis. An incidental diagnosis in a Saudi female.

Dracunculiasis is a disabling, and economically crippling parasitic infestation transmitted by drinking contaminated water. Although the disease has been eradicated from most parts of the world, it is still endemic in some tropical African countries. Here we report a 65-year-old female from the southern region of Saudi Arabia with radiological evidence of heavy load of guinea worms. This case could represent the local reemergence of the disease.