Use of quantitative PCR to assess the efficacy of albendazole against Necator americanus and Ascaris spp. in Manufahi District, Timor-Leste.

BACKGROUND: Soil-transmitted helminths (STHs) including Ascaris lumbricoides, Necator americanus, Ancylostoma spp. and Trichuris trichiura are cause of significant global morbidity. To mitigate their disease burden, at-risk groups in endemic regions receive periodic mass drug administration using anthelmintics, most commonly albendazole and mebendazole. Assessing the efficacy of anthelmintic drugs is important for confirming that these regimens are working effectively and that drug resistance has not emerged. In this study we aimed to characterise the therapeutic efficacy of albendazole against Ascaris spp. and N. americanus in Timor-Leste, using a quantitative polymerase chain reaction (qPCR) method for parasite detection and quantification. RESULTS: A total of 314 participants from 8 communities in Timor-Leste provided stool samples before and 10-14 days after the administration of a single 400 mg dose of albendazole. Helminth infection status and infection intensity (measured in Ct-values and relative fluorescence units) were determined using qPCR. Efficacy was determined by examining the cure rates and infection intensity reduction rates. Albendazole was found to be highly efficacious against Ascaris spp., with a cure rate of 91.4% (95% CI: 85.9-95.2%) and infection intensity reduction rate of 95.6% (95% CI: 88.3-100%). The drug was less efficacious against N. americanus with a cure rate of 58.3% (95% CI: 51.4-64.9%) and infection intensity reduction rate of 88.9% (95% CI: 84.0-97.0%). CONCLUSIONS: The observed cure rates and infection intensity reduction rates obtained for Ascaris spp. and to a lower extent N. americanus, demonstrate the continued efficacy of albendazole against these species and its utility as a mass chemotherapy agent in Timor-Leste. Furthermore, this study demonstrates the usefulness of qPCR as a method to measure the efficacy of anthelminthic drugs. Additional research is necessary to translate Ct-values into eggs per gram in a systematic way. TRIAL REGISTRATION: Australian and New Zealand Clinical Trials Registry 12614000680662 (registered 27 June 2014).

The assessment of hookworm calreticulin as a potential vaccine for necatoriasis.

A vaccine against the human hookworm Necator americanus is urgently required to reduce hookworm-induced morbidity in endemic areas. In the present study, recombinant hookworm calreticulin, a nominated vaccine candidate, has been tested in mice. Mice given calreticulin had 43-49% fewer worms in their lungs, compared to non-vaccinated controls, following challenge infection with infective hookworm larvae. These levels of protection were achieved in the absence of adjuvant following intraperitoneal administration of three doses of 15 microg antigen. Antigen was also encapsulated in PLG microparticles. Encapsulated calreticulin elicited higher levels of anti-calreticulin IgG1 than free antigen but failed to induce protective immunity. The protection induced by free calreticulin was associated with low levels of serum IgE and moderate lung eosinophilia whilst administration of calreticulin-loaded microparticles was associated with high levels of serum IgE and higher lung eosinophil activity, suggesting that the classical Th2 phenotype may not always be associated with protective immunity, albeit in experimental necatoriasis.

Hookworm disease: host-pathogen biology.

The hallmark of hookworm infection, a common helminthic disease, is iron deficiency anemia. The development of anemia is dependent on the intensity of infection, the species of hookworm, and the ability of the host to resist infection and to maintain adequate stores of iron. When conditions are appropriate, the incidence of anemia caused by hookworm is high and has a significant economic impact since it results in a reduction of worker productivity. Loss of blood is caused by direct ingestion of red cells and by tissue trauma produced by worm attachment and feeding. This focal trauma may involve multiple villi and is characterized by local hemorrhage, tissue cytolysis, and neutrophilic response. Although focal intestinal lesions are apparent, their significance is questionable since diffuse mucosal changes are absent in intestinal biopsies of patients with heavy hookworm infection. Short-range control measures protecting against hookworm infection have not succeeded. Development of a vaccine against hookworm infection in humans is problematic since functional protective immunity in humans has not yet been demonstrated and no suitable animal model of hookworm infection in humans is available. At present, the most effective method of intervention appears to be supplementation of food staples with iron.

Hookworm disease: nutritional implications.

Iron-deficiency anemia resulting from intestinal blood loss is the major consequence of hookworm infection. Development of the anemia can be prevented, and it can be treated by administration of iron. Hypoproteinemia, often associated with hookworm infection, may be the result of either protein malnutrition or increased intestinal loss of protein. It is unlikely that the worms cause diffuse morphologic or functional alterations of the intestine. Fortification or supplementation with iron is a practical method to control hookworm disease in endemic areas.