Development of an in vitro technique for cytological investigations of slices of Fasciola hepatica: Evaluation by physiological criteria

1975 ◽  
Vol 5 (3) ◽  
pp. 333-337 ◽  
Author(s):  
L.T. Threadgold ◽  
B. Hanna
Keyword(s):  
Fasciola Hepatica ◽  
In Vitro Technique

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 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 Transcriptomic analysis reveals a role for the nervous system in regulating growth and development of Fasciola hepatica juveniles

2022 ◽  
Author(s):  
Emily Robb ◽  
Erin McCammick ◽  
Duncan Wells ◽  
Paul McVeigh ◽  
Erica Gardiner ◽  
...  
Keyword(s):  
Nervous System ◽  
Rapid Growth ◽  
Growth And Development ◽  
Liver Fluke ◽  
Fasciola Hepatica ◽  
Expression Profiles ◽  
Neuronal Signalling ◽  
Growth Development

Fasciola spp. liver fluke have significant impacts in veterinary and human medicine. The absence of a vaccine and increasing anthelmintic resistance threaten sustainable control and underscore the need for novel flukicides. Functional genomic approaches underpinned by in vitro culture of juvenile Fasciola hepatica facilitate control target validation in the most pathogenic life stage. Comparative transcriptomics of in vitro and in vivo maintained 21 day old F. hepatica finds that 86% of genes are expressed at similar levels across maintenance treatments suggesting commonality in core biological functioning within these juveniles. Phenotypic comparisons revealed higher cell proliferation and growth rates in the in vivo juveniles compared to their in vitro counterparts. These phenotypic differences were consistent with the upregulation of neoblast-like stem cell and cell-cycle associated genes in in vivo maintained worms. The more rapid growth/development of in vivo juveniles was further evidenced by a switch in cathepsin protease expression profiles, dominated by cathepsin B in in vitro juveniles and then by cathepsin L in in vivo juveniles. Coincident with more rapid growth/development was the marked downregulation of both classical and peptidergic neuronal signalling components in in vivo maintained juveniles, supporting a role for the nervous system in regulating liver fluke growth and development. Differences in the miRNA complements of in vivo and in vitro juveniles identified 31 differentially expressed miRNAs, notably fhe-let-7a-5p , fhe-mir-124-3p and, miRNAs predicted to target Wnt-signalling, supporting a key role for miRNAs in driving the growth/developmental differences in the in vitro and in vivo maintained juvenile liver fluke. Widespread differences in the expression of neuronal genes in juvenile fluke grown in vitro and in vivo expose significant interplay between neuronal signalling and the rate of growth/development, encouraging consideration of neuronal targets in efforts to dysregulate growth/development for parasite control.

An in vitro technique for surveying immunostimulants in fish

Aquaculture ◽  
1993 ◽  
Vol 112 (2-3) ◽  
pp. 283-287 ◽  
Author(s):  
Galina Jeney ◽  
Douglas P. Anderson
Keyword(s):  
In Vitro Technique
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