Optimization and workflow of in vitro culture of adult Fasciola hepatica.

IF 1.3 4区农林科学 Q2 Veterinary

Revista Brasileira De Parasitologia Veterinaria Pub Date : 2024-11-22 eCollection Date: 2024-01-01 DOI:10.1590/S1984-29612024064

Cesar Burga-Cisterna, Edith Málaga, Enrique Serrano-Martínez, Giovana Livia-Córdova, Ramón Antezana, Américo Castro Luna, Monica Pajuelo

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Abstract

The aim of this study was to evaluate different transportation and incubation conditions to test the viability of adult Fasciola hepatica in order to propose a new cultivation workflow. The adult stage of F. hepatica was obtained from naturally infected cattle at a local slaughterhouse in Lima, Peru. Different transport and incubation conditions of F. hepatica were tested, evaluating its viability through a motility scale. DMEM and RPMI 1640 media presented better transport conditions compared to Hedon-Fleig and PBS media (p < 0.001), maintaining the flukes at 37°C. Also, DMEM and RPMI-1640 media presented better incubation conditions compared to Hedon-fleig (p < 0.001). A minimum of 3 ml of medium per fluke was required to maintain best viability (p < 0.001) and no differences in viability were found between the different types of culture plates (p > 0.05). In addition, we found that incubation with DMSO (dimethyl sulfoxide) at concentrations greater than 0.5% v/v for 48 hours generates toxicity (p < 0.001). In conclusion, RPMI 1640 and DMEN media presented better transport and in vitro cultivation conditions for F. hepatica, using DMSO at concentrations lower than 0.5% v/v.

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肝吸虫成虫体外培养的优化和工作流程。

本研究的目的是评估不同的运输和培养条件，以测试肝包虫成虫的存活率，从而提出一种新的培养工作流程。肝吸虫成虫是从秘鲁利马当地屠宰场自然感染的牛身上获得的。对肝包虫的不同运输和培养条件进行了测试，并通过运动量表评估其活力。与 Hedon-Fleig 和 PBS 培养基相比，DMEM 和 RPMI 1640 培养基的运输条件更好（p < 0.001），能将吸虫保持在 37°C。此外，DMEM 和 RPMI-1640 培养基的孵育条件也优于 Hedon-Fleig（p < 0.001）。每条吸虫至少需要 3 毫升培养基才能保持最佳的存活率（p < 0.001），不同类型的培养板在存活率上没有差异（p > 0.05）。此外，我们发现用浓度大于 0.5% v/v 的 DMSO（二甲基亚砜）培养 48 小时会产生毒性（p < 0.001）。总之，使用浓度低于 0.5% v/v 的二甲基亚砜，RPMI 1640 和 DMEN 培养基能为肝蝇提供更好的运输和体外培养条件。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Revista Brasileira De Parasitologia Veterinaria PARASITOLOGY-VETERINARY SCIENCES

CiteScore

2.60

自引率

7.70%

发文量

审稿时长

>12 weeks

期刊介绍： La revista es un órgano de difusión del Colegio Brasileño de Parasitología Veterinaria, con una especificidad dentro de esa área, la difusión de los resultados de la investigación brasileña en las áreas de Helmintología, Protozoología, Entomología y agentes transmitidos por artrópodos, relacionados con la salud animal.