Coelomic fluid aspiration has been utilized in echinoderms in research and clinical settings. Detailed procedural descriptions for coelomic fluid sampling in sea urchins (class Echinoidea) are lacking, and samples are prone to contamination. The objectives of this study were to (1) standardize a technique for coelomic fluid collection in long-spined sea urchin Diadema antillarum that optimizes the diagnostic quality of the sample utilizing diagnostic imaging, (2) identify coelomic fluid bacterial isolates (using Biolog GEN III MicroLog and 16s rDNA sequencing), and (3) compare positive cultures to animal weight, holding time prior to sampling, water temperature, and gross fluid appearance. Seventy Diadema antillarum from the Florida Keys collected in two groups (March and September 2015) were utilized. Positive cultures for bacterial contamination were identified in 5% and 44%, respectively, of animals in the sampling groups. Vibrio spp. was the predominant genus identified. Positive cultures were more frequent in the group with smaller-sized animals, increased holding times, and elevated water temperatures. Deviation from clear-pink gross coelomic fluid appearance did not reliably predict bacterial contamination. A standardized technique for coelomocentesis was defined. The use of the proposed coelomocentesis methodology may facilitate improved health evaluations of sea urchins and may be applicable to research, conservation efforts, and disease investigations.

中文翻译：

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建立长刺海胆Diadema antillarum体腔穿刺术的诊断技术

体腔液抽吸已在研究和临床环境中用于棘皮动物。海胆（棘皮纲）体腔液取样的详细程序描述缺乏，并且样品容易受到污染。本研究的目的是 (1) 标准化一种在长刺海胆 Diadema antillarum 中收集体腔液的技术，该技术利用诊断成像优化样本的诊断质量，(2) 鉴定体腔液细菌分离物（使用 Biolog GEN III MicroLog 和 16s rDNA 测序），以及 (3) 将阳性培养物与动物体重、采样前的保持时间、水温和总液体外观进行比较。使用了来自佛罗里达礁岛群的 70 粒 Diadema antillarum，分为两组（2015 年 3 月和 9 月）。分别在 5% 和 44% 的采样组动物中鉴定出细菌污染的阳性培养物。弧菌属 是确定的优势属。在体型较小、饲养时间增加和水温升高的组中，阳性培养更频繁。与透明粉红色大体体腔液外观的偏差不能可靠地预测细菌污染。定义了体腔穿刺术的标准化技术。使用拟议的体腔穿刺方法可能有助于改进海胆的健康评估，并可能适用于研究、保护工作和疾病调查。在体型较小、饲养时间增加和水温升高的组中，阳性培养更为频繁。与透明粉红色大体体腔液外观的偏差不能可靠地预测细菌污染。定义了体腔穿刺术的标准化技术。使用拟议的体腔穿刺方法可能有助于改进海胆的健康评估，并可能适用于研究、保护工作和疾病调查。在体型较小、饲养时间增加和水温升高的组中，阳性培养更频繁。与透明粉红色大体体腔液外观的偏差不能可靠地预测细菌污染。定义了体腔穿刺术的标准化技术。使用拟议的体腔穿刺方法可能有助于改进海胆的健康评估，并可能适用于研究、保护工作和疾病调查。