水蛭壶菌游动孢子活力测定方法的比较。

IF 1.1 4区 农林科学 Q3 FISHERIES Diseases of aquatic organisms Pub Date : 2023-09-14 DOI:10.3354/dao03749
Adeline Chew, Matt West, Laura A Brannelly
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引用次数: 0

摘要

新型真菌病原体水蛭壶菌(Batrachochytrium dendroatidis, Bd)威胁着全球数百种两栖动物。在以实验室为基础的实验中,通常必须对活的Bd细胞进行量化,但对计数和评估感染性游动孢子生命阶段活力的方法的有效性进行比较尚未完成。直接比较评估细胞活力的染色方法将确保使用最准确和有效的方法。在这里,我们比较了两种相对便宜的常用染色剂,台盼蓝和亚甲基蓝,并评估了它们在人工计数和比色测定中估计Bd游动孢子活力的准确性和精密度。我们用每种染色剂对已知比例的bdzoo孢子(0、0.25、0.50、0.75和1.00)进行染色,并使用人工计数和比色法估计每个样品中染色(死亡)和未染色(活)细胞的比例。台盼蓝被发现是比亚甲基蓝更有效的染色显微镜和比色测定。此外,通过显微镜计算游动孢子是一种更准确和精确的技术。我们建议使用人工计数,通过显微镜使用台盼蓝染色来评估Bd游动孢子的活力。
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Comparison of methods for determining Batrachochytrium dendrobatidis zoospore viability.

The emerging fungal pathogen Batrachochytrium dendrobatidis (Bd) threatens hundreds of amphibian species globally. During laboratory-based experiments it is often essential to quantify live Bd cells, but a comparison of the effectiveness of methods for counting and assessing the viability of the infectious zoospore life stage has not been done. A direct comparison of staining methods that assess viability will ensure that the most accurate and efficient method is used. Here, we compared the use of 2 relatively cheap common stains, trypan blue and methylene blue, and assessed their accuracy and precision for estimating the viability of Bd zoospores during both manual counting and colorimetric assays. We stained known proportions of killed Bd zoospores (0, 0.25, 0.50, 0.75, and 1.00) with each stain and estimated the proportion of stained (dead) and unstained (viable) cells in each sample using both manual counting and colorimetric assays. Trypan blue was found to be a much more effective stain than methylene blue for both microscopy and colorimetric assays. Additionally, counting zoospores via microscopy was both a more accurate and precise technique. We recommend using manual counts via microscopy using the trypan blue stain for assessing Bd zoospore viability.

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来源期刊
Diseases of aquatic organisms
Diseases of aquatic organisms 农林科学-兽医学
CiteScore
3.10
自引率
0.00%
发文量
53
审稿时长
8-16 weeks
期刊介绍: DAO publishes Research Articles, Reviews, and Notes, as well as Comments/Reply Comments (for details see DAO 48:161), Theme Sections and Opinion Pieces. For details consult the Guidelines for Authors. Papers may cover all forms of life - animals, plants and microorganisms - in marine, limnetic and brackish habitats. DAO''s scope includes any research focusing on diseases in aquatic organisms, specifically: -Diseases caused by coexisting organisms, e.g. viruses, bacteria, fungi, protistans, metazoans; characterization of pathogens -Diseases caused by abiotic factors (critical intensities of environmental properties, including pollution)- Diseases due to internal circumstances (innate, idiopathic, genetic)- Diseases due to proliferative disorders (neoplasms)- Disease diagnosis, treatment and prevention- Molecular aspects of diseases- Nutritional disorders- Stress and physical injuries- Epidemiology/epizootiology- Parasitology- Toxicology- Diseases of aquatic organisms affecting human health and well-being (with the focus on the aquatic organism)- Diseases as indicators of humanity''s detrimental impact on nature- Genomics, proteomics and metabolomics of disease- Immunology and disease prevention- Animal welfare- Zoonosis
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