Medinensis Infection Research Articles

Identifying correlates of Guinea worm (Dracunculus medinensis) infection in domestic dog populations.

Few human infectious diseases have been driven as close to eradication as dracunculiasis, caused by the Guinea worm parasite (Dracunculus medinensis). The number of human cases of Guinea worm decreased from an estimated 3.5 million in 1986 to mere hundreds by the 2010s. In Chad, domestic dogs were diagnosed with Guinea worm for the first time in 2012, and the numbers of infected dogs have increased annually. The presence of the parasite in a non-human host now challenges efforts to eradicate D. medinensis, making it critical to understand the factors that correlate with infection in dogs. In this study, we evaluated anthropogenic and environmental factors most predictive of detection of D. medinensis infection in domestic dog populations in Chad. Using boosted regression tree models to identify covariates of importance for predicting D. medinensis infection at the village and spatial hotspot levels, while controlling for surveillance intensity, we found that the presence of infection in a village was predicted by a combination of demographic (e.g. fishing village identity, dog population size), geographic (e.g. local variation in elevation), and climatic (e.g. precipitation and temperature) factors, which differed between northern and southern villages. In contrast, the presence of a village in a spatial infection hotspot, was primarily predicted by geography and climate. Our findings suggest that factors intrinsic to individual villages are highly predictive of the detection of Guinea worm parasite presence, whereas village membership in a spatial infection hotspot is largely determined by location and climate. This study provides new insight into the landscape-scale epidemiology of a debilitating parasite and can be used to more effectively target ongoing research and possibly eradication and control efforts.

Ecology of domestic dogs Canis familiaris as an emerging reservoir of Guinea worm Dracunculus medinensis infection.

Global eradication of human Guinea worm disease (dracunculiasis) has been set back by the emergence of infections in animals, particularly domestic dogs Canis familiaris. The ecology and epidemiology of this reservoir is unknown. We tracked dogs using GPS, inferred diets using stable isotope analysis and analysed correlates of infection in Chad, where numbers of Guinea worm infections are greatest. Dogs had small ranges that varied markedly among villages. Diets consisted largely of human staples and human faeces. A minority of ponds, mostly <200 m from dog-owning households, accounted for most dog exposure to potentially unsafe water. The risk of a dog having had Guinea worm was reduced in dogs living in households providing water for animals but increased with increasing fish consumption by dogs. Provision of safe water might reduce dog exposure to unsafe water, while prioritisation of proactive temephos (Abate) application to the small number of ponds to which dogs have most access is recommended. Fish might have an additional role as transport hosts for Guinea worm, by concentrating copepods infected with worm larvae.

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.

A Woman with Knee Pain and Soft-Tissue Calcification

The patient reported a history of having had a thread-like worm at 10-12 years of age (i.e., >50 years before presentation). At that time, she was a resident of Najran, Saudi Arabia, and recalled experiencing intense itching in the lower limbs, followed by the emergence of a small worm that her family removed by wrapping it around a stick. Since then, she had experienced no associated problems. Radiographs of the patient's knees revealed guinea worm calcifications. Serpentine soft-tissue calcification is seen within the distal aspect of the right thigh and around the left knee (figure 1). The calcification appears nodular, beaded, and fragmented because of the breaking up of the worm (figure 2). The characteristic appearance, combined with the patient's background, led to a diagnosis of infection due to D. medinensis (guinea-worm), a nematode parasite that causes dracunculiasis. After the death of an adult worm, D. medinensis may undergo calcification in several forms, especially if it was not fertilized or did not release its larvae. In its typical location in the lower extremities, the female D. medinensis appears as a long, stringlike, serpiginous or curvilinear calcification, which may extend to 1 meter in length [1]. A description of D. medinensis infection was given by ArRazi Al Hawi several hundred years ago [2]. The disease is a debilitating and painful infection that begins with a blister, usually on the leg, followed by itching, fever, swelling, and burning sensations. Once the infected person immerses the infected part in open water, the worms emerge and release thousands of larvae into the water. The larva is then ingested by a water flea (cyclops), where it develops and becomes infective. Humans acquire infection by drinking water contaminated with cyclops. The D. medinensis larva then penetrates the gut wall and migrates through the subcutaneous tissue,

Antibody and cytokine responses in Dracunculus medinensis patients at distinct states of infection

Dracunculiasis is a promising candidate for eradication, but transmission of Dracunculus medinensis and recrudescence of the disease have been observed repeatedly. In the present investigation, the D. medinensis-specific cellular cytokine response profiles and the parasite-specific antibody subclass reactivity were evaluated in dracunculiasis patients at distinct states of infection. Analysis of the cellular cytokine response in dracunculiasis patients disclosed a D. medinensis antigen-specific depression of IFN-gamma production with patent D. medinensis infection, while the T helper type 2 cytokine IL-10 was similar in patent, post-patent and control individuals, and IL-5 production was always the highest in controls. In parallel, diminished IFN-gamma and IL-12 responses to antigens from Ascaris lumbricoides, Entamoeba histolytica and mycobacteria were observed in patent and post-patent dracunculiasis cases. The parasite-specific IgG(1) and IgG(4) subclass reactivity profiles corresponded with the D. medinensis infection state, and a clear distinction between patent and post-patent patients and controls was found. Overall a depressed cytokine release was observed with patent D. medinensis, which extended beyond the parasite-specific immune responsiveness. The detection of D. medinensis-specific IgG(1) and IgG(4) isotypes may help to distinguish newly exposed, patent and post-patent D. medinensis infections.

Human serum albumin and immunoglobulin on Dracunculus medinensis

Dracunculus medinensis recovered from infected humans were examined for the occurrence and localization of human serum albumin and immunoglobulins. Immunoelectrophoretic examination of homogenates prepared from adult female worms (ADGW) and first stage larvae (LVGW) showed that anti-human albumin antibodies reacted to both stages of the parasite. By direct fluorescence antibody technique, antigens resembling human albumin and human immunoglobulins (isotype IgG) were identified on the surface of adult female worms. The occurrence of host-like compounds on the parasites may be an adaptation for survival of the parasites. A possible interference of these compounds should be considered in attempts to develop methods for immunological diagnosis of D. medinensis infections.

Immunoepidemiology of Dracunculus medinensis infections II. Variation in antibody responses in relation to transmission season and patency.

The serum antibody responses (specific IgG1, IgG4, and IgE, and total IgE) to Dracunculus medinensis infection in humans from a highly endemic area of northern Ghana were examined regularly by ELISA over a period of one year in cohorts of individuals who developed a patent D. medinensis infection during the study period (actively infected category), or who claimed to have never had a patent infection (endemic normal category). The results were analyzed in relation to seasonality and time of patency of infection. For individuals in the actively infected category, a clear seasonal variation in the mean levels of specific IgG1 and IgG4 was found, with the highest levels late in the dry season and early in the rainy season, when transmission is high, and the lowest levels late in the rainy season and early in the dry season. Endemic normal individuals responded with low and fluctuating levels of specific IgG1 and with low and nonfluctuating levels of specific IgG4. For specific and total IgE, no seasonal variation was observed in any of the two infection status categories. In relation to time of patency of infection (only involving the category of actively infected individuals), the mean levels of specific IgG1 and IgG4 increased from two months before patency of infection, peaked during patency, and then gradually decreased for four months until a constant level was reached. No significant fluctuations in the levels of specific and total IgE were observed in relation to time of patency. The present study thus showed extensive variation in levels of D. medinensis-specific IgG1 and IgG4 (but not IgE) over time. Seasonal variations in antibody responses may also occur in other helminth infections, especially those with seasonal transmission, and these should be taken into consideration when interpreting the results of immunologic studies.

Immunoepidemiology of Dracunculus medinensis infections I. Antibody responses in relation to infection status.

The specific serum IgG1, IgG4, and IgE responses to Dracunculus medinensis and the level of total IgE of individuals living in a highly endemic area of northern Ghana were measured by ELISA. Sera were obtained in the high transmission season from individuals with prepatent, patent, or postpatent infection as well as from individuals from the same endemic area who claimed to have never had a patent infection (i.e., endemic normal individuals). Individuals with prepatent or postpatent infections responded with a significantly lower mean level of specific IgG1 and IgG4 compared with individuals with a patent infection, and with a significantly higher mean level of specific IgG1 and IgG4 compared with endemic normal individuals. For specific IgE, no differences were found in the mean antibody level between the infection status categories. Individuals with a patent infection had a significantly lower mean serum level of total IgE compared with prepatent, postpatent, and endemic normal individuals. Endemic normal individuals had the highest mean level of total IgE. Furthermore, in all clinical categories, high responders for specific IgG1 and IgG4 generally had low levels of total IgE, whereas low responders for specific IgG1 and IgG4 generally had high levels of total IgE. A similar dichotomy, although less distinct, was observed between specific IgG1 and IgG4 on the one hand and specific IgE on the other. Thus, similar to what has been suggested for schistosomiasis and lymphatic filariasis, the relationship between the IgG subclasses and IgE appears to play a role in, or at least to reflect, a mechanism for protective immunity in dracunculiasis.

The significance of guinea worm infection in the immunological diagnosis of onchocerciasis and bancroftian filariasis

Infections with Dracunculus medinensis frequently occur in the same geographical area as infections with Onchocerca volvulus and Wuchereria bancrofti. This study analysed the significance of D. medinensis infections for the specificity and sensitivity of available tests for antibody-based diagnosis of onchocerciasis (using individual recombinant clones OV-10, OV-11 and OV-16, and the OV-7/OV-10/OV-16 tri-cocktail, in an enzyme-linked immunosorbent assay) and for circulating antigen-based diagnosis of bancroftian filariasis (using the TroBio ™ and the ICT ™ card tests). Some immunological cross-reactivity was observed with all tests. When using individual recombinant O. volvulus antigens, the highest assay indices were obtained for clone OV-10, and the lowest for clone OV-16. Testing the serum responses against the tri-cocktail of recombinant antigens did not notably improve the assay indices. Two of 40 serum samples from individuals with patent dracunculiasis gave a false positive response in the ICT ™ test and one of these was also positive in the TropBio ™ test. Possible implications of applying these diagnostic assays in areas endemic for dracunculiasis are discussed.

Studies on immunodiagnosis of dracunculiasis. I. Detection of specific serum antibodies

To explore the possibility for development of an immunodiagnostic test for Dracunculus medinensis infections, the antibody responses (IgG1, IgG2, IgG3, IgG4 and IgE) to antigen preparations made from adult female worms (ADGW) and first stage larvae (LVGW) of D. medinensis were analysed. By sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE)/Western blotting, sera from individuals with patent D. medinensis infection reacted extensively and similarly with ADGW and LVGW over a broad molecular weight range (5–200 kDa). Sera from individuals infected with Onchocerca volvulus (the major cross-reacting infection) had a more pronounced reaction with ADGW than LVGW, and only with antigens of molecular weights above 23 kDa. These findings were used to design an enzyme-linked immunosorbent assay (ELISA) with high sensitivity and specificity for D. medinensis infection. The most promising results were obtained when detecting for specific IgG4. Thus, when using LVGW, the assay had a sensitivity of 83% and a specificity of 97%. Refining the antigen into a low molecular weight filtrate improved the sensitivity to 92%, and the sensitivity was further improved (to 96%) when combining the results from two or more antibody types measured simultaneously, without affecting the specificity. The results were encouraging for the prospects for developing an applicable immunodiagnostic test for patent D. medinensis infections based on detection of specific serum antibodies.

Immunodiagnosis of dracunculiasis by dot-ELISA.

A dot-enzyme-linked immunosorbent assay (dot-ELISA) using soluble adult-guineaworm antigen was developed for immunodiagnosis of Dracunculus medinensis infection. The test was found to be specific and sensitive when compared with ELISA. The dot-ELISA described here was found to be time-saving and easy to perform, and its possible use as a diagnostic and epidemiological tool under field conditions is therefore discussed.

Immunodiagnosis of Dracunculiasis by Falcon Assay Screening Test-Enzyme-Linked Immunosorbent Assay (Fast-Elisa) and by Enzyme-Linked Immunoelectrotransfer Blot (EITB) Technique

The Falcon assay screening test-enzyme-linked immunosorbent assay (FAST-ELISA) and the enzyme-linked immunoelectrotransfer blot (EITB) technique were used to test human sera with Dracunculus medinensis adult worm antigen in order to assess their potential value in the immunodiagnosis of dracunculiasis. The human sera used were from patients with prepatent and patent D. medinensis infections or from patients infected with other nematodes (Onchocerca volvulus and Loa loa) or trematodes (Schistosoma mansoni and S. haematobium), as well as uninfected Nigerian and Puerto Rican normal controls. In the FAST-ELISA, the sera from prepatent and patent dracunculiasis patients gave the highest absorbance values relative to normal human sera. The highest cross-reactivity was observed with onchocerciasis sera; no cross-reactivity was seen with sera from individuals with loiasis or schistosomiasis mansoni or haematobia. By the EITB, sera from dracunculiasis patients specifically recognized a 16 kDa protein (Dm 16) and antibodies to Dm 16 disappeared 2 months after worm extraction. Recognition of Dm 16 occurred from the late prepatent stage. A 17 kDa protein (Dm 17) was also recognized by dracunculiasis sera, but antibodies to Dm 17 disappeared more slowly and were present 1 year after recovery. The 16 kDa and 17 kDa antigens of D. medinensis may be useful in the immunodiagnosis of dracunculiasis.