Supercapacitance/Resistance Behaviors of Helminth Eggs as Reliable Recognition and Direct Differentiation Probe

Parasitic helminths are usually known as undesired pathogens, causing various diseases in both human and animal species. In this study, we explore supercapacitance/resistance behaviors as a novel probe for rapid identification and direct differentiation of Fasciola hepatica, Parascaris equorum (with and without larvae), Dicrocoelium dendriticum, Taenia multiceps, and Moniezia expansa eggs. This claim is attributed to some characteristics, such as grave supercapacitance/area, high-energy storage/area, large power/egg, huge permittivity, and great electrical break-down potential, respectively (Fasciola hepatica: 2,158, 0.485, 2.7 × 10–3, 267, 52.6, Parascaris equorum without larvae: 2,825, 0.574, 3.0 × 10–3, 351, 68.4, Parascaris equorum with larvae: 4,519, 0.716, 2.4 × 10–3, 1.96, 97.6, Dicrocoelium dendriticum: 1,581, 0.219, 2.8 × 10–3, 1.96, 48.8, Moniezia expansa: 714, 0.149, 2.2 × 10–3, 0.88, 35.2, Taenia multiceps: 3,738, 0.619, 4.7 × 10–3, 4.63, 84.4), and durable capacitance up to at least 15,000 sequential cycles at different scan rates (between 2.0 × 10−4 and 120.0 V s−1) as well as highly differentiated resistance between 400 and 600 Ω. These traits are measured by the “Blind Patch-Clamp” method, at the giga ohm sealed condition (6.18 ± 0.12 GΩ cm−1, n = 5). Significant detection ranges are detected for each capacitance and resistance with gradient limits as large as at least 880 to 1,000 mF and 400 to 600 Ω depending on the type of helminth egg. The effect of water in the structure of helminth eggs has also been investigated with acceptable reproducibility (RSD 7%–10%, n = 5). These intrinsic characteristics would provide novel facilitators for direct helminth egg identification in comparison with several methods, such as ELISA, PCR, and microscopic methods.

In Vitro Assessment of Anthelmintic Activities of AgO Nanoparticle in Comparison to Closantel Against Liver Fluke Dicrocoelium Dendriticum

Background: Dicrocoeliasis is a rare Food-Born parasitic disease of the grazing herbivores as well humans, caused by Dicrocoelium dendriticum making severe pathological changes of the liver and bile systems, and therapeutic options for treatment are limited. With the appearance of drug resistance in liver flukes, there is a need to focus on alternative approaches to control helminth parasites of veterinary importance. Because of low-performance medications; drug delivery poses a great challenge for better treatment of Dicrocoeliasis. The current study aims to determine the anthelmintic properties of silver oxide nanoparticles (AgO) as a new method in dicrocoeliasis treatment, in vitro assay.Methods: The impacts of various concentrations of AgO nanoparticles (50-200 µg/ml) for 12-24 hours were compared with the Closantel, as the chemical drug. The anthelmintic efficacy was measured by scanning electron microscopy (SEM) technique.Results: SEM images of treated worms by AgO (200 µg/ml) showed severe damage, including complete loss of sensory papillae and destruction of prominent network structures and tegument vesicles. The mortality rates how the anthelmintic properties of AgO were highly relied on time and concentration, as far as increasing the time and concentration cause increasing the mortality rate. According to the MTT assay, the toxicity of AgO, at concentrations, 800 µg/ml is 8.7 %. Conclusions: Hence, it could be concluded that AgO NPs performed anthelmintic properties effects. To the best of our knowledge, no previous reports have assessed the effect of AgO NP on liver fluke D.dendriticum. Therefore, the present study provides a basis for future research on the control of this common trematode.

Immunopathological Response, Histological Changes, Parasitic Burden, and Egg Output in Sheep Naturally Infected by Dicrocoelium dendriticum

The aim of this study was to investigate the correlation between infection by Dicrocoelium dendriticum (class Trematoda) and the animal host response in terms of macroscopic lesions, the immunopathological response, and histological changes in the livers of naturally infected sheep. Twenty-four sheep were selected on the basis of positive D. dendriticum fecal egg counts (FECs). Gross and histological injuries were scored. A positive significant association was observed between the number of adult worms recovered from the liver, FEC, macroscopic lesions, fibrosis, and bile duct hyperplasia. A significant negative association was observed among these variables and the degree of leukocyte infiltration. In addition, immunophenotyping of the inflammatory cells was carried out using primary antibodies against T cell epitopes (CD3+, CD4+, and CD8+), B cell epitopes (CD79α), and the ionized calcium-binding adapter molecule 1 (IBA-1) antigen. Independently of the severity of the D. dendriticum infection, the predominant cell population was CD3-positive and associated with lesser numbers of CD79α- and Iba-I-positive cells. An increase in Iba-1-positive cells was observed in the livers of animals with a high worm burden. Our results provide a reference basis to better understand the local immune response in sheep naturally infected by D. dendriticum in relation to the FEC and parasitic burden.

Molecular characterization and immunodiagnostics of Dicrocoelium dendriticum species isolated from sheep of north-west Himalayan region

Despite its extensive presence among grazing ruminants, dicrocoeliosis, also known as ‘small liver fluke’ disease, is poorly known and often underestimated by researchers and practitioners in many countries. The accurate identification and prepatent diagnosis of Dicrocoelium dendriticum infection is an essential prerequisite for its prevention and control. In the present study, the morphologically identified specimens isolated from the bile ducts of sheep (Ovis aries) were validated through molecular data. The sequence analysis of the second internal transcribed spacer (ITS-2) of our isolates showed a high degree of similarity with D. dendriticum using the BLAST function of the National Center for Biotechnology Information (NCBI). The phylogenetic analysis of our isolates showed a close relationship with previously described D. dendriticum isolates from different countries. The antigenic profiles of somatic and excretory/secretory (E/S) antigens of D. dendriticum were revealed by sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS–PAGE) and immunoblotting using sera from sheep naturally infected with D. dendriticum. By SDS–PAGE, 16 distinct bands were revealed from crude somatic fraction. Immunoblotting analysis of these proteins with positive sera exhibited six seroreactive bands ranging from 27 to 130 kDa. Among these, the 84 and 130 kDa bands were quite specific, with high diagnostic specificity and sensitivity. The E/S fraction comprised nine distinct bands, as revealed by SDS–PAGE analysis. Immunoblotting analysis of these proteins with positive sera exhibited five antigenic bands ranging from 27 to 130 kDa. Among these, the 130 kDa band was found to be quite specific, with high diagnostic specificity and sensitivity. The present study concludes that the protein bands of 84 and 130 kDa in somatic fraction and 130 kDa in E/S fraction can be used for the immunodiagnostic purpose for this economically important parasite, which may also encourage further studies regarding their vaccine potential.

Morphological Characteristics of Dicrocoelium dendriticum (Digenea, Dicrocoeliidae), Parasitizing Three Host Species in the Central Regions of Ukraine

Distribution of trematodes of the species Dicrocoelium dendriticum (Rudolphi, 1819) Looss, 1899 (Digenea: Dicrocoeliidae) parasitizing cattle (Bos taurus Linnaeus, 1758), sheep (Ovis aries Linnaeus, 1758) and goat (Capra aegagrus hircus Linnaeus, 1758) was studied in the Poltava and Kirovohrad regions of Ukraine. The recorded prevalence of infection is 26.9 % in cattle, 28.42 % in sheep and 24.5 % in goat. D. dendriticum is the only species of the genus Dicrocoelium which infects domestic animals in the central regions of Ukraine. The trematodes obtained from cattle are significantly different from those isolated from sheep and goat by length, width and area of body, and length of vitelline ducts.