Virus and prokaryote abundances from experiments conducted with samples collected at a hydrothermal vent site by ROV SuBastian during R/V Falkor (too) expedition FKt230627 along the East Pacific Rise in July of 2023

We used the virus-dilution technique to quantify lytic virus production at nine sites across vent, sub-vent, and non-vent habitats along the East Pacific Rise during expedition FKt230627 aboard R/V Falkor(too). This technique leverages the density-dependent nature of viral infection, preventing new infections by dilution. Samples were collected via ROV SuBastian, sequentially filtered to remove larger particles, and concentrated using tangential-flow filtration. Duplicate incubations were set up with prokaryotic concentrate and virus-free water, pressurized to 250 bar, and incubated in the dark at in situ temperatures for 30 hours. Subsamples were taken every six hours for enumeration of prokaryotes and viruses via flow cytometry. Parallel experiments were conducted at surface pressure to assess the impact of pressure on virus production rates. The methodology ensures observed increases in viral abundance are due to pre-existing infections. The experiments were conducted aboard R/V Falkor(too) between 5-22 July 2023 by Tinkara Tinta and Nicole Krause. Prokaryotes and viruses were enumerated flow-cytometrically back in the lab by Christian Winter.

在R/V Falkor(too)科考船执行FKt230627航次期间，本研究采用病毒稀释技术对东太平洋海脊沿线喷口、亚喷口及非喷口栖息地的9个站点的裂解性病毒产量进行了定量分析。该技术利用病毒感染的密度依赖性特征，通过稀释抑制新感染的发生。样本由遥控潜水器（ROV）SuBastian采集，经逐级过滤去除大颗粒后，采用切向流过滤（tangential-flow filtration）浓缩。将原核生物浓缩液与无病毒水混合设置平行培养组，加压至250巴，并在原位温度、黑暗条件下培养30小时。每6小时采集一次子样本，通过流式细胞术（flow cytometry）对原核生物和病毒进行计数。同时在表面压力条件下开展平行实验，以评估压力对病毒产生速率的影响。该方法确保观测到的病毒丰度增加源于已存在的感染。实验由Tinkara Tinta和Nicole Krause于2023年7月5日至22日在R/V Falkor(too)科考船上完成；原核生物和病毒的流式细胞计数由Christian Winter在实验室完成。