Genetic detection and discrimination of closely related oyster pathogens for biosecurity and shellfish stock management.

IF 1.1 4区 农林科学 Q3 FISHERIES
Jaret P Bilewitch, Henry S Lane, Kathryn H Wiltshire, Marty R Deveney, Amber R Brooks, Keith P Michael
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引用次数: 0

Abstract

Disease outbreaks among shellfish cause significant declines in both wild fisheries and aquaculture production globally. In Aotearoa New Zealand, a long history of flat oyster infections by the haplosporidian parasite Bonamia exitiosa has influenced fishery dynamics in the Foveaux Strait. B. ostreae, which causes high host mortality, was detected in both wild and farmed oysters in the Marlborough Sounds in 2015 and at Stewart Island in 2017, prompting the development of rapid diagnostic tests with high sensitivity and specificity that can be used to simultaneously monitor both Bonamia species. Separate molecular diagnostic assays were developed for B. exitiosa and B. ostreae using droplet digital PCR (ddPCR). Analytical tests indicated both assays were specific and had limits of detection below 0.4 copies µl-1. Estimates of diagnostic performance used latent class analysis conducted on comparisons with previously established test methods. Both ddPCR assays had a diagnostic sensitivity and specificity equal to or greater than tests based on qPCR, end-point PCR, histopathology and heart imprints, validating their use for fishery health assessment and biosecurity monitoring. The new ddPCR assays are preferred over other methods in situations where high throughput and analytic and diagnostic performance are essential, as seen in New Zealand where 2 Bonamia species are sympatric in some regions and may co-infect a single host. Furthermore, the new assays could be adapted for eDNA-based surveillance and combined with other pathogen assays for multiplex assays, enabling further research into Bonamia lifecycles.

近缘牡蛎病原菌的遗传检测与鉴定及其在生物安全和贝类种群管理中的应用。
贝类疾病的爆发导致全球野生渔业和水产养殖产量显著下降。在新西兰的奥特罗阿,单孢子虫寄生虫博纳米亚寄生的扁平牡蛎感染的悠久历史影响了福沃克斯海峡的渔业动态。2015年在马尔伯勒海峡和2017年在斯图尔特岛的野生和养殖牡蛎中都发现了导致高宿主死亡率的B. ostreae,这促使开发了具有高灵敏度和特异性的快速诊断测试,可用于同时监测两种博纳米亚物种。采用液滴数字PCR (ddPCR)技术分别建立了出口双歧杆菌和牛链球菌的分子诊断方法。分析试验表明,这两种方法都具有特异性,检测限低于0.4拷贝µl-1。估计诊断性能使用潜类分析进行比较与先前建立的测试方法。两种ddPCR检测方法的诊断敏感性和特异性均等于或高于基于qPCR、终点PCR、组织病理学和心脏印记的检测方法,验证了它们在渔业健康评估和生物安全监测方面的应用。在高通量和分析诊断性能至关重要的情况下,新的ddPCR检测比其他方法更受欢迎,例如在新西兰,2种博纳米亚原虫在某些地区是同域的,可能同时感染一个宿主。此外,新的检测方法可以适用于基于edna的监测,并与其他病原体检测方法结合进行多重检测,从而进一步研究博纳米亚菌的生命周期。
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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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