Ciesielski Mark, Clerkin Thomas, Funnell Nicholas, Ben-Horin Tal, Noble Rachel T
Department of Marine Sciences, Institute of Marine Sciences (IMS), University of North Carolina at Chapel Hill, Morehead City, North Carolina, USA.
Department of Veterinary Medicine, North Carolina State University Center for Marine Sciences and Technology (CMAST), North Carolina State University, Morehead City, North Carolina, USA.
Appl Environ Microbiol. 2025 Apr 23;91(4):e0214924. doi: 10.1128/aem.02149-24. Epub 2025 Apr 2.
The shellfish aquaculture industry is one of the fastest-growing sectors of global food production, but it is currently facing major challenges stemming from microbial pathogens. This study presents an optimized and validated suite of droplet digital PCR (ddPCR) assays using water samples proximal to oyster farms in North Carolina to quantify pathogens relevant to the aquaculture industry. Two of the molecular assays enable the quantification of the pathogens, and , that threaten human health and oyster performance, respectively. This work also introduces two ddPCR assays that enable the simultaneous quantification of at least nine ecologically relevant . using only two sets of primers and probes targeting the glycosyl hydrolase family 18 (GH18) domain of the gene in bacteria. The entire suite of assays was applied to single assessments at 12 sites, revealing heterogeneity in microbial pathogen concentrations across the coastal landscape and variability of abundances within individual estuarine river systems. Additionally, a longitudinal study conducted at a demonstration lease elucidated unique temporal trends for all microbial targets. Notably, when concentrations of . quantified using the two assays targeting the gene reached their maximum, the daily probability of mortality increased, suggesting a role for other ecologically pertinent . in the progression of mortality that would otherwise be missed. This study highlights the utility of ddPCR for the advancement of shellfish management by offering insights into the spaciotemporal dynamics of microbial pathogens.
Climate change is drastically altering the environment and changing the abundance and geographical distribution of marine pathogens. These microbial species put additional pressure on the aquaculture industry by acting as sources of disease for animals important to the food industry as well as for humans upon consumption of contaminated food. To address growing concerns, high-resolution monitoring of pathogens can offer insights for effective management in a critical industry. Validated in the field, the suite of molecular droplet digital PCR assays presented here improves upon current methods, enabling the simultaneous quantification of several targets. This technology makes it possible to track pathogens as they move through the environment and reveals changes in abundance that may inform adjustments to farming practices aimed at mitigating negative outcomes. Additionally, this work presents a unique approach to molecular assay design that unveils potential drivers of ecological shifts and emerging etiologies of disease more efficiently.
贝类水产养殖业是全球粮食生产中发展最快的行业之一，但目前正面临着微生物病原体带来的重大挑战。本研究提出了一套经过优化和验证的液滴数字PCR（ddPCR）检测方法，使用北卡罗来纳州牡蛎养殖场附近的水样来量化与水产养殖业相关的病原体。其中两种分子检测方法能够分别对威胁人类健康和牡蛎生长性能的病原体 和 进行定量。这项工作还引入了两种ddPCR检测方法，仅使用两组针对 细菌中 基因糖基水解酶家族18（GH18）结构域的引物和探针，就能同时对至少九种与生态相关的 进行定量。整套检测方法应用于对12个地点的单次评估，揭示了沿海地区微生物病原体浓度的异质性以及各个河口河流系统内丰度的变异性。此外，在一个示范租赁区进行的纵向研究阐明了所有微生物目标独特的时间趋势。值得注意的是，当使用针对 基因的两种检测方法定量的 浓度达到最大值时，每日死亡率增加，这表明其他与生态相关的 在死亡率进展中起到了作用，否则这些作用会被忽视。本研究通过深入了解微生物病原体的时空动态，突出了ddPCR在推进贝类管理方面的实用性。
气候变化正在急剧改变环境，改变海洋病原体的丰度和地理分布。这些微生物物种给水产养殖业带来了额外压力，它们既是对食品行业重要的动物的疾病来源，也是人类食用受污染食物时的疾病来源。为了应对日益增长的担忧，对病原体进行高分辨率监测可为这个关键行业的有效管理提供见解。本文介绍的这套经过现场验证的分子液滴数字PCR检测方法改进了现有方法，能够同时对多个目标进行定量。这项技术使追踪病原体在环境中的移动成为可能，并揭示丰度变化，这些变化可为旨在减轻负面结果的养殖实践调整提供依据。此外，这项工作提出了一种独特的分子检测设计方法，能更有效地揭示生态变化的潜在驱动因素和新出现的疾病病因。
