Corrigendum to "scFv against HSP60 of Strongyloides sp. and Its Application in the Evaluation of Parasite Frequency in the Elderly".

[This corrects the article DOI: 10.1155/2020/4086929.].

Innovative approaches to improve serodiagnosis of Strongyloides stercoralis infection.

Strongyloidiasis is a helminthiasis of neglected condition that has no gold standard parasitological diagnosis due to the intermittent release of larvae in feces. This study aimed to use an scFv (single chain variable fragment) obtained by Phage Display, previously validated to detect immune complexes in serum samples from individuals infected with Strongyloides stercoralis by enzyme-linked immunosorbent assay (ELISA). Now the ability of scFv to detect the immune complexes was verified by immunofluorescence, flow cytometry using magnetic beads and surface plasmon resonance (SPR). As ELISA, the SPR, immunofluorescence and flow cytometry demonstrated the ability of scFv to detect immune complexes in sera from individuals with strongyloidiasis and discriminate them from sera of individuals with other parasitic diseases and healthy individuals. Besides de conventional ELISA, the novel approaches can also be promptly applied as auxiliary diagnostic tools to the existing parasitological method for accurate diagnosis of human strongyloidiasis.

scFv against HSP60 of Strongyloides sp. and Its Application in the Evaluation of Parasite Frequency in the Elderly.

The present study is aimed at evaluating serological method using scFv anti-Strongyloides sp. and reporting the frequencies of the results with conventional parasitological technique (faeces) in elderly individuals. Among 112 elderly individuals (≥60 years of age), 14.28% were positive for at least one enteroparasite, with one individual positive for S. stercoralis. Sera were evaluated for the presence of anti-Strongyloides sp. antibodies using total or detergent fraction extracts of Strongyloides venezuelensis, which presented positivity rates of 19.64% and 10.71%, respectively. An anti-HSP60 single-chain variable fragment from Strongyloides sp. was used to detect parasite antigens, with 5.36% (6 individuals) of ELISA-positive individuals returning a positive result. While the serological test indicates previous or recent infection and may be limited by antigen purification, the anti-HSP60 method reflects the presence of Strongyloides sp. immune complexes and exhibits greater sensitivity and specificity. Our results demonstrate the variable occurrence of enteroparasites in elderly individuals residing in long-term nursing homes and validate a novel epidemiological tool to describe infection cases by Strongyloides sp.

In vitro ovicidal and larvicidal activity of Carica papaya seed hexane extract against Strongyloides venezuelensis.

Strongyloidiasis is a human parasitic disease caused by the helminth Strongyloides stercoralis whose treatment is particularly difficult in immunosuppressed patients due to their low responsiveness to conventional therapy. Carica papaya and its isolated compounds benzyl isothiocyanate, carpaine and carpasemine are promising compound for the treatment of Strongyloides infections due to their anthelmintic action. This study aims to examine the in vitro ovicidal and larvicidal activity of C. papaya seed hexane extract against Strongyloides venezuelensis, using egg hatching tests and larval motility tests as efficiency markers. The crude extract at the concentrations of 566 - 0.0566 mg/mL or the control with albendazole (0.025 mg/mL) and negative controls (water and PBS) were incubated with an equal volume of egg suspension (± 50 specimens) followed by counting of the specimens after 48 h. The same extract and dilutions were added to L3 larvae suspensions (±50 specimens) followed by analysis of larvae viability after 24, 48, and 72 h. The extract inhibited egg hatching with high efficiency at concentrations of 56.6 mg/mL (95.74%) and 5.66 mg/mL (92.16%). At the concentrations of 566 mg/mL (100%) and 56.66 mg/mL (97.32%), the extract inhibited larval motility as effectively as ivermectin (0.316 mg/mL; 100%), and more effectively than the other dilutions and the negative controls. The larvicidal effect depended on the extract concentration, but not on the treatment period. Therefore, C. papaya seed hexane extract has anthelmintic potential against S. venezuelensis and is a promising compound for the development of phytotherapies to treat strongyloidiasis.

In vitro efficacy of latex and purified papain from Carica papaya against Strongyloides venezuelensis eggs and larvae.

Latex from Carica papaya is rich in bioactive compounds, especially papain, which may help to control parasitic diseases. This study evaluated the efficacy of latex from C. papaya and purified papain against Strongyloides venezuelensis. The Egg Hatching Test (EHT) and the Larval Motility Test (LMT) using fresh and frozen latex (250mg/mL), lyophilized latex (34mg/mL), and purified papain (2.8 mg/mL) were performed. Albendazole (0.025 mg/mL) and ivermectin (316 ppm) were used as positive controls. EHT and LMT were carried out through the incubation of each solution with S. venezuelensis eggs or larvae (± 100 specimens), and results were analyzed after 48h (EHT) or 24, 48, and 72h (LMT). EHT showed that latex preparations at higher concentrations (1:10 to 1:100) resulted in partial or complete destruction of eggs and larvae inside the eggs. The result from the 1:1,000 dilution was similar to the positive control. LMT showed effectiveness in all the tested dilutions compared to negative controls. Purified papain showed a dose-dependent response in the EHT. Purified papain (2.8 mg/ mL) showed similar results to lyophilized latex at 1:1,000 in the EHT. Latex and purified papain from C. papaya were effective against S. venezuelensis eggs and larvae in vitro, suggesting their potential use as an alternative treatment for strongyloidiasis.

In vitro efficacy of latex and purified papain from Carica papaya against Strongyloides venezuelensis eggs and larvae

ABSTRACT Latex from Carica papaya is rich in bioactive compounds, especially papain, which may help to control parasitic diseases. This study evaluated the efficacy of latex from C. papaya and purified papain against Strongyloides venezuelensis. The Egg Hatching Test (EHT) and the Larval Motility Test (LMT) using fresh and frozen latex (250mg/mL), lyophilized latex (34mg/mL), and purified papain (2.8 mg/mL) were performed. Albendazole (0.025 mg/mL) and ivermectin (316 ppm) were used as positive controls. EHT and LMT were carried out through the incubation of each solution with S. venezuelensis eggs or larvae (± 100 specimens), and results were analyzed after 48h (EHT) or 24, 48, and 72h (LMT). EHT showed that latex preparations at higher concentrations (1:10 to 1:100) resulted in partial or complete destruction of eggs and larvae inside the eggs. The result from the 1:1,000 dilution was similar to the positive control. LMT showed effectiveness in all the tested dilutions compared to negative controls. Purified papain showed a dose-dependent response in the EHT. Purified papain (2.8 mg/ mL) showed similar results to lyophilized latex at 1:1,000 in the EHT. Latex and purified papain from C. papaya were effective against S. venezuelensis eggs and larvae in vitro, suggesting their potential use as an alternative treatment for strongyloidiasis.

Current progress toward vaccine and passive immunization approaches for Strongyloides spp.

Strongyloides stercoralis is a helminth parasite that can infect millions of people worldwide, particularly in tropical, subtropical and temperate regions with poor sanitation. Several aspects of epidemiology, biology and host-parasite interactions of S. stercoralis have been studied, and substantial knowledge has been acquired; however, very few studies on immunotherapeutic control strategies to prevent infection and disease in humans have been conducted. Therefore, this article reviews the current progress and targets toward vaccine and passive immunization approaches for Strongyloides spp.

Structural and functional characterization of a novel scFv anti-HSP60 of Strongyloides sp.

Phage display is a powerful technology that selects specific proteins or peptides to a target. We have used Phage Display to select scFv (single-chain variable fragment) clones from a combinatorial library against total proteins of Strongyloides venezuelensis. After scFv characterization, further analysis demonstrated that this recombinant fragment of antibody was able to bind to an S. venezuelensis antigenic fraction of ~65 kDa, present in the body periphery and digestive system of infective larvae (L3), as demonstrated by immunofluorescence. Mass spectrometry results followed by bioinformatics analysis showed that this antigenic fraction was a heat shock protein 60 (HSP60) of Strongyloides sp. The selected scFv was applied in serodiagnosis by immune complexes detection in serum samples from individuals with strongyloidiasis using a sandwich enzyme-linked immunosorbent assay (ELISA), showing sensitivity of 97.5% (86.84-99.94), specificity of 98.81 (93.54-99.97), positive likelihood ratio of 81.60 and an area under the curve of 0.9993 (0.9973-1.000). Our study provided a novel monoclonal scFv antibody fragment which specifically bound to HSP60 of Strongyloides sp. and was applied in the development of an innovative serodiagnosis method for the human strongyloidiasis.

The detergent fraction is effective in the detection of IgG anti-Strongyloides stercoralis in serum samples from immunocompromised individuals.

Human strongyloidiasis is an intestinal helminthiasis that can be fatal particularly in cases of immunosuppression. The aim of this study is to assess the diagnostic accuracy of the detergent fraction (D), purified from total saline extract (SE) of Strongyloides venezuelensis, in the detection of anti-Strongyloides stercoralis IgG antibodies in serum samples from individuals coming from endemic areas for strongyloidiasis and presenting immunocompromised conditions: human immunodeficiency virus (HIV(+)), diabetes mellitus type 2, cancer, tuberculosis and alcoholism. Serum samples from 93 individuals were analyzed by ELISA, as follows: Group 1: 30 immunocompromised individuals with strongyloidiasis; Group 2: 33 immunocompromised individuals without strongyloidiasis and Group 3: 30 healthy individuals. The total saline extract (SE) and detergent fraction (D) showed a sensitivity of 73.33 and 83.33%, and specificity of 82.15 and 86.36%, respectively. The detergent fraction was effective to detect anti-S. stercoralis IgG antibodies in immunocompromised individuals with strongyloidiasis and may be applied as an important tool in the immunodiagnosis of human strongyloidiasis related to immunosuppression.

The role of cytoskeleton, components of inositol phospholipid signaling pathway and iron in Ehrlichia canis in vitro proliferation.

Ehrlichia canis, etiologic agent of Canine Monocytic Ehrlichiosis, is an obligatory intracellular bacterium that parasitizes monocytes and macrophages. In this study we analyzed the role of the cytoskeleton specifically actin microfilaments and microtubules, components of inositol phospholipid signaling pathway such as phospholipase C (PLC), protein kinase (PTK) and calcium channels as well as the role of iron in the E. canis proliferation in DH82 cells. Different inhibitory compounds were used for each component: Cytochalasin D (inhibits actin polymerization), Nocodazole (inhibits microtubule polymerization), Neomycin (PLC inhibitor), Genistein (PTK inhibitor), Verapamil (calcium channel blocker) and Deferoxamine (iron chelator). We observed a significant decrease in the total number of bacteria in infected cells treated suggesting that these cellular components analized are essentials to E. canis proliferation.