Water column bacteria data from ARSV Laurence M. Gould cruises LMG0104 and LMG0106 in the Southern Ocean in 2001 (SOGLOBEC project; Sea Ice Microbes project)

<h2>Bacteria Abundance, Biomass and Chlorophyll: Water Column Samples<em>a</em></h2>

<p><strong>BG 235 - Methods used for chlorophyll <em>a</em> (chla) analysis and bacteria biomass determination</strong></p>

<p><strong>Core Sampling techniques:</strong></p>

<p>Sampling methods for recovery of chlorophyll <em>a</em> and bacteria from sea ice cores follows those described in&nbsp;<br />
Garrison and Buck (1986)</p>

<p>Recommendations for reporting were used as outlined by:&nbsp;Horner, R. et al.,(1992)</p>

<p><strong>Analytic Techniques:</strong></p>

<p>Chla (mg m^-3):</p>

<ul>
<li>determined fluorometrically (Turner Designs 10AU Fluorometer) following extraction in 90% acetone (Parsons <em>et al</em>., 1984)</li>
<li>ice core chla corrected to account for chla in filtered sea water (FSW) added to core sections during melting</li>
</ul>

<p>Bacteria cell abundance (cells m^-3) and biomass (mg C m^-3):</p>

<p><strong>LMG 0106</strong></p>

<ul>
<li>preserved (0.5% glutaraldehyde) samples stained with 4',6-diamidino-2-phenylindole (DAPI; 0.1 to 0.3% final concentration), filtered through 0.2 mm black, polycarbonate membrane filters, and mounted onto glass microscope slides on the ship (within 24 hours following collection)</li>
<li>bacteria enumerated using epifluorescence microscopy and sized using digital images taken with Image Pro Plus</li>
<li>bacteria biomass determined using cell abundance, cell biovolume (BV; mm³; as determined from mean length and width measurements), and an allometric conversion factor for bacterial carbon per volume specific for DAPI-stained bacteria (cellular carbon = 218 X BV^0.86; Loferer-Krossbacher <em>et al.</em>, 1998).</li>
<li>ice core samples corrected for FSW dilution</li>
</ul>

<p><strong>NBP 0104</strong></p>

<ul>
<li>preserved (0.5% glutaraldehyde) samples stained with SYBR Gold (0.01% final concentration), filtered through 0.2 mm Anodisc filters (Whatman), and mounted onto glass microscope slides at home institution (~1-2 months following collection)</li>
<li>bacteria enumerated using epifluorescence microscopy and sized using digital images taken with Image Pro Plus</li>
<li>bacteria biomass determined using cell abundance, cell biovolume (BV; mm³), and an allometric conversion factor for bacterial carbon per volume specific for Acridine Orange-stained bacteria (cellular carbon = 89.9 X BV^0.59; Simon and Azam, 1989). Note: an AO-specific carbon per volume conversion factor was used in calculating biomass in SYBR Gold-stained samples because both AO and SYBR Gold stain bacteria cells similarly relative to DAPI (unpublished data).</li>
<li>ice core samples corrected for FSW dilution</li>
</ul>

<p>Data from <strong>LMG0106</strong> (July-August, 2001) added in June 2002.</p>

<p><em>Updated: April 21, 2006</em></p>

<h2>细菌丰度、生物量与叶绿素：水柱样本<em>a</em></h2>

<p><strong>BG 235 — 叶绿素<em>a</em>（chla）分析与细菌生物量测定方法</strong></p>

<p><strong>岩芯采样技术：</strong></p>

<p>从海冰冰芯中提取叶绿素<em>a</em>与细菌的采样方法，遵循Garrison与Buck（1986）所述的实验方案。</p>

<p>数据报告规范参考Horner R.等人（1992）提出的指南。</p>

<p><strong>分析技术：</strong></p>

<p>叶绿素a（chla，单位：mg·m⁻³）：</p>

<ul>
<li>采用荧光光度法（使用Turner Designs 10AU型荧光光度计）测定，样品经90%丙酮萃取（Parsons等，1984）</li>
<li>对冰芯叶绿素a含量进行校正，以扣除融解冰芯过程中加入的过滤海水（filtered sea water，缩写FSW）中所含的叶绿素a</li>
</ul>

<p>细菌细胞丰度（单位：cells·m⁻³）与生物量（单位：mg C·m⁻³）：</p>

<p><strong>LMG 0106</strong></p>

<ul>
<li>样品经0.5%戊二醛固定后，用4',6-二脒基-2-苯基吲哚（4',6-diamidino-2-phenylindole，缩写DAPI；终浓度0.1%~0.3%）染色，通过0.2 mm黑色聚碳酸酯膜过滤，采样后24小时内于科考船操作台制片于载玻片上</li>
<li>采用落射荧光显微镜对细菌进行计数，并通过Image Pro Plus获取的数字图像测量细菌尺寸</li>
<li>细菌生物量通过细胞丰度、细胞生物体积（biovolume，缩写BV；单位：mm³，由细胞平均长、宽测量值计算得到），以及针对DAPI染色细菌的专属体积-细菌碳质量异速转换因子计算得到（细胞碳含量=218×BV^0.86；Loferer-Krossbacher等，1998）</li>
<li>冰芯样本校正值扣除FSW稀释效应</li>
</ul>

<p><strong>NBP 0104</strong></p>

<ul>
<li>样品经0.5%戊二醛固定后，用SYBR Gold（终浓度0.01%）染色，通过0.2 mm Anodisc滤膜（Whatman品牌）过滤，采样后1~2个月内于所属研究机构制片于载玻片上</li>
<li>采用落射荧光显微镜对细菌进行计数，并通过Image Pro Plus获取的数字图像测量细菌尺寸</li>
<li>细菌生物量通过细胞丰度、细胞生物体积（BV；单位：mm³），以及针对吖啶橙（Acridine Orange，缩写AO）染色细菌的专属体积-细菌碳质量异速转换因子计算得到（细胞碳含量=89.9×BV^0.59；Simon与Azam，1989）。注：由于SYBR Gold与吖啶橙（AO）对细菌细胞的染色效果相较于DAPI更为相近（未发表数据），因此在计算SYBR Gold染色样本的生物量时，采用了针对AO染色的体积-碳质量转换因子</li>
<li>冰芯样本校正值扣除FSW稀释效应</li>
</ul>

<p><strong>LMG0106</strong>批次的采样数据（2001年7-8月）于2002年6月补充入库。</p>

<p><em>更新日期：2006年4月21日</em></p>