In vitro maintenance of Fasciola hepatica: A factorial approach based on egg production

1969 ◽  
Vol 26 (1) ◽  
pp. 41-51 ◽  
Author(s):  
Leslie H Ractliffe ◽  
Diego Guevara-Pozo ◽  
Ramon Lopez-Roman
Keyword(s):  
Egg Production ◽  
Fasciola Hepatica ◽  
Factorial Approach ◽  
In Vitro Maintenance

scholarly journals Targeting Kinases in Fasciola hepatica: Anthelminthic Effects and Tissue Distribution of Selected Kinase Inhibitors

2020 ◽  
Vol 7 ◽  
Author(s):  
Carolin M. Morawietz ◽  
Hicham Houhou ◽  
Oliver Puckelwaldt ◽  
Laura Hehr ◽  
Domenic Dreisbach ◽  
...  
Keyword(s):  
Tissue Distribution ◽  
Protein Kinases ◽  
Egg Production ◽  
Kinase Inhibitors ◽  
Fasciola Hepatica ◽  
Human Cancer ◽  
Oral Route ◽  
New Drugs ◽  
Uptake Kinetic

Protein kinases have been discussed as promising druggable targets in various parasitic helminths. New drugs are also needed for control of fascioliasis, a food-borne trematode infection and worldwide spread zoonosis, caused by the liver fluke Fasciola hepatica and related species. In this study, we intended to move protein kinases more into the spotlight of Fasciola drug research and characterized the fasciolicidal activity of two small-molecule inhibitors from human cancer research: the Abelson tyrosine kinase (ABL-TK) inhibitor imatinib and the polo-like 1 (PLK1) inhibitor BI2536. BI2536 reduced viability of 4-week-old immature flukes in vitro, while adult worms showed a blockade of egg production. Together with a significantly higher transcriptional expression of PLK1 in adult compared to immature worms, this argues for a role of PLK1 in fluke reproduction. Both fluke stages expressed ABL1-TK transcripts at similar high levels and were affected by imatinib. To study the uptake kinetic and tissue distribution of imatinib in F. hepatica, we applied matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) for the first time in this parasite. Drug imaging revealed the accumulation of imatinib in different fluke tissues from 20 min to 12 h of exposure. Furthermore, we show that imatinib is metabolized to N-desmethyl imatinib by F. hepatica, a bioactive metabolite also found in humans. Besides the vitellarium, gastrodermal tissue showed strong signal intensities. In situ hybridization demonstrated the gastrodermal presence of abl1 transcripts. Finally, we assessed transcriptional changes of physiologically important genes in imatinib-treated flukes. Moderately increased transcript levels of a gene encoding a multidrug resistance protein were detected, which may reflect an attempt to defend against imatinib. Increased expression levels of the cell cycle dependently expressed histone h2b and of two genes encoding superoxide dismutases (SODs) were also observed. In summary, our pilot study demonstrated cross-stage activity of imatinib but not BI2536 against immature and adult F. hepatica in vitro; a fast incorporation of imatinib within minutes, probably via the oral route; and imatinib-induced expression changes of physiologically relevant genes. We conclude that kinases are worth analyzing in more detail to evaluate the potential as therapeutic targets in F. hepatica.

scholarly journals Comparative proteomic profiling of newly acquired, virulent and attenuated Neoparamoeba perurans proteins associated with amoebic gill disease

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Kerrie Ní Dhufaigh ◽  
Eugene Dillon ◽  
Natasha Botwright ◽  
Anita Talbot ◽  
Ian O’Connor ◽  
...  
Keyword(s):  
Bacterial Community Composition ◽  
Illumina Miseq ◽  
Proteomic Profiling ◽  
Farmed Fish ◽  
Liquid Chromatography Tandem Mass ◽  
Proteome Database ◽  
Gill Disease ◽  
The Impact ◽  
In Vitro Maintenance

AbstractThe causative agent of amoebic gill disease, Neoparamoeba perurans is reported to lose virulence during prolonged in vitro maintenance. In this study, the impact of prolonged culture on N. perurans virulence and its proteome was investigated. Two isolates, attenuated and virulent, had their virulence assessed in an experimental trial using Atlantic salmon smolts and their bacterial community composition was evaluated by 16S rRNA Illumina MiSeq sequencing. Soluble proteins were isolated from three isolates: a newly acquired, virulent and attenuated N. perurans culture. Proteins were analysed using two-dimensional electrophoresis coupled with liquid chromatography tandem mass spectrometry (LC–MS/MS). The challenge trial using naïve smolts confirmed a loss in virulence in the attenuated N. perurans culture. A greater diversity of bacterial communities was found in the microbiome of the virulent isolate in contrast to a reduction in microbial community richness in the attenuated microbiome. A collated proteome database of N. perurans, Amoebozoa and four bacterial genera resulted in 24 proteins differentially expressed between the three cultures. The present LC–MS/MS results indicate protein synthesis, oxidative stress and immunomodulation are upregulated in a newly acquired N. perurans culture and future studies may exploit these protein identifications for therapeutic purposes in infected farmed fish.

scholarly journals The soluble glutathione transferase superfamily: role of Mu class in triclabendazole sulphoxide challenge in Fasciola hepatica

2021 ◽  
Vol 120 (3) ◽  
pp. 979-991
Author(s):  
Rebekah B. Stuart ◽  
Suzanne Zwaanswijk ◽  
Neil D. MacKintosh ◽  
Boontarikaan Witikornkul ◽  
Peter M. Brophy ◽  
...  
Keyword(s):  
In Vitro Culture ◽  
Liver Fluke ◽  
Glutathione Transferase ◽  
Fasciola Hepatica ◽  
Affinity Purification ◽  
Economic Loss ◽  
Differential Abundance ◽  
Parasitic Helminths

AbstractFasciola hepatica (liver fluke), a significant threat to food security, causes global economic loss for the livestock industry and is re-emerging as a foodborne disease of humans. In the absence of vaccines, treatment control is by anthelmintics; with only triclabendazole (TCBZ) currently effective against all stages of F. hepatica in livestock and humans. There is widespread resistance to TCBZ and its detoxification by flukes might contribute to the mechanism. However, there is limited phase I capacity in adult parasitic helminths with the phase II detoxification system dominated by the soluble glutathione transferase (GST) superfamily. Previous proteomic studies have demonstrated that the levels of Mu class GST from pooled F. hepatica parasites respond under TCBZ-sulphoxide (TCBZ-SO) challenge during in vitro culture ex-host. We have extended this finding by exploiting a sub-proteomic lead strategy to measure the change in the total soluble GST profile (GST-ome) of individual TCBZ-susceptible F. hepatica on TCBZ-SO-exposure in vitro culture. TCBZ-SO exposure demonstrated differential abundance of FhGST-Mu29 and FhGST-Mu26 following affinity purification using both GSH and S-hexyl GSH affinity. Furthermore, a low or weak affinity matrix interacting Mu class GST (FhGST-Mu5) has been identified and recombinantly expressed and represents a new low-affinity Mu class GST. Low-affinity GST isoforms within the GST-ome was not restricted to FhGST-Mu5 with a second likely low-affinity sigma class GST (FhGST-S2) uncovered. This study represents the most complete Fasciola GST-ome generated to date and has supported the potential of subproteomic analyses on individual adult flukes.

scholarly journals High-resolution AP-SMALDI MSI as a tool for drug imaging in Schistosoma mansoni

2021 ◽  
Author(s):  
Annika S. Mokosch ◽  
Stefanie Gerbig ◽  
Christoph G. Grevelding ◽  
Simone Haeberlein ◽  
Bernhard Spengler
Keyword(s):  
Schistosoma Mansoni ◽  
Egg Production ◽  
Drug Repurposing ◽  
Cancer Drug ◽  
Ionization Mass ◽  
Parasitic Flatworm ◽  
Paired Female ◽  
First Time ◽  
Organ Tropism

AbstractSchistosoma mansoni is a parasitic flatworm causing schistosomiasis, an infectious disease affecting several hundred million people worldwide. Schistosomes live dioeciously, and upon pairing with the male, the female starts massive egg production, which causes pathology. Praziquantel (PZQ) is the only drug used, but it has an inherent risk of resistance development. Therefore, alternatives are needed. In the context of drug repurposing, the cancer drug imatinib was tested, showing high efficacy against S. mansoni in vitro. Besides the gonads, imatinib mainly affected the integrity of the intestine in males and females. In this study, we investigated the potential uptake and distribution of imatinib in adult schistosomes including its distribution kinetics. To this end, we applied for the first time atmospheric-pressure scanning microprobe matrix-assisted laser desorption/ionization mass spectrometry imaging (AP-SMALDI MSI) for drug imaging in paired S. mansoni. Our results indicate that imatinib was present in the esophagus and intestine of the male as early as 20 min after in vitro exposure, suggesting an oral uptake route. After one hour, the drug was also found inside the paired female. The detection of the main metabolite, N-desmethyl imatinib, indicated metabolization of the drug. Additionally, a marker signal for the female ovary was successfully applied to facilitate further conclusions regarding organ tropism of imatinib. Our results demonstrate that AP-SMALDI MSI is a useful method to study the uptake, tissue distribution, and metabolization of imatinib in S. mansoni. The results suggest using AP-SMALDI MSI also for investigating other antiparasitic compounds and their metabolites in schistosomes and other parasites. Graphical abstract

scholarly journals Inmunidad Celular en Ganado Vacuno Lechero Naturalmente Infectado con Fasciola hepatica en Cajamarca, Perú

2015 ◽  
Vol 26 (4) ◽  
pp. 637
Author(s):  
Heber Silva-Díaz ◽  
Cristian Hobán-Vergara ◽  
Rosmery Cruz-Cerna ◽  
Hugo Solana ◽  
Pedro Ortiz-Oblitas
Keyword(s):  
Fasciola Hepatica ◽  
Ifn Γ

<p>La fasciolosis producida por <em>Fasciola hepatica</em> es una importante enfermedad parasitaria de la ganadería en muchos países. En Cajamarca, Perú, se reportan prevalencias superiores al 80% en el ganado lechero. El objetivo del estudio fue evaluar la capacidad de proliferación y la expresión de citoquinas (IFN-γ e IL-4) en células mononucleares de sangre periférica de terneras y vacas naturalmente infectadas, contra el antígeno no específico fitohemaglutinina y los antígenos específicos de excreción-secreción del estadio inmaduro y maduro de <em>F. hepatica</em>. La capacidad proliferativa fue determinada por linfoproliferación <em>in vitro</em> y la expresión de citoquinas se evaluó en sobrenadantes de cultivo celular por la técnica de ELISA. Las vacas infectadas con <em>F. hepatica</em> mostraron una disminución de la capacidad de respuesta frente a estímulos proliferativos inespecíficos y específicos. En el perfil de citoquinas contra el estímulo específico se encontró una baja expresión de IFN-γ, mientras que la respuesta de IL-4 fue alta, lo que indica que la respuesta permanece polarizada hacia una respuesta de tipo Th2. En el análisis comparativo, las terneras expresaron niveles de IFN-γ más altos que los valores obtenidos en las vacas, pero con una expresión similar de IL-4. No se observaron diferencias en la respuesta inmune a los antígenos del estadio inmaduro y maduro del parásito. Los resultados demuestran que a medida que la enfermedad progresa se expresa una respuesta inmunomoduladora en los animales adultos naturalmente infectados con <em>F. hepatica</em>.</p>

scholarly journals In vitro effects of artesunate on the survival of worm pairs and egg production of Schistosoma mansoni

2009 ◽  
Vol 83 (1) ◽  
pp. 7-11 ◽  
Author(s):  
Y. Mitsui ◽  
M. Miura ◽  
Y. Aoki
Keyword(s):  
Schistosoma Mansoni ◽  
Adult Worm ◽  
Survival Time ◽  
Egg Production ◽  
Inhibitory Effect ◽  
Male And Female ◽  
Paired Male ◽  
In Vitro Effects ◽  
Paired Female

AbstractThe effect of artesunate (ART) on the survival time of adult worm pairs of Schistosoma mansoni and on their egg output during in vitro culture was assessed. ART significantly decreased the survival time of both paired male and female worms at concentrations of 5, 10, 20 and 40 mg l− 1 during in vitro cultivation. An inhibitory effect of ART on the daily egg output of paired female worms during in vitro cultivation was also observed.

The effects of Fasciola hepatica on Lymnaea truncatula

Parasitology ◽  
1972 ◽  
Vol 65 (2) ◽  
pp. 359-369 ◽  
Author(s):  
J. K. M. Hodasi
Keyword(s):  
Rapid Growth ◽  
Egg Production ◽  
Fasciola Hepatica ◽  
Age Groups ◽  
Sharp Decline ◽  
General Belief ◽  
Functional Changes ◽  
Complete Cessation ◽  
Lymnaea Truncatula ◽  
Accelerated Growth

Among the functional changes observed in L. truncatula infected with F. hepatica were those of growth, reproduction and mortality. For the first 3 weeks after infection there was no difference between the growth of infected and uninfected groups of snails. Thereafter there was a temporary accelerated growth which occurred between the fourth and seventh weeks after infection. The onset of this period of rapid growth in the different age groups of infected snails coincided with the time of migration of the rediae into the hepatopancreas. Of more significance was the influence of infection on the reproduction of the snail. There was a sharp decline in oviposition 2–3 weeks after infection, followed by a complete cessation of egg production at the end of the fifth week. Once oviposition ceased, it was not resumed and there was no self-cure. Contrary to general belief, infected groups of L. truncatula have been found to live longer than the uninfected controls. With isolated snails, however, there was a tendency for the uninfected snails to live longer than the infected ones. The possible interpretation of the discrepancies between isolated and mass cultured snails are discussed. The hepatopancreas and the gonad (ovotestis) were the two structures directly affected by the infection. Establishment of the rediae in the hepatopancreas invariably resulted in parasitic castration.

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