Microinvertebrates Survey on Macquarie Island

MICROINVERTEBRATE SAMPLING PROTOCOLMacquarie Island01 October 2001 - 28 February 2002A.HABITATS SAMPLED8 habitats representative of the following vegetation types were chosen:1.Azorella macquariensis - Open cushion areas2.Acaena (magellanica and minor) herbfield3.Colobanthus muscoides (coastal cushion plants)4.Mires - Upland5.Pleurophyllum hookerii dominated areas6.Poa foliosa Tall tussock7.Short grassland (incl. Agrostis magellanica/ Festuca contracta/ Luzula)8.Stilbocarpa polaris dominated coastal herbfieldB.HABITAT LOCALITIES1.Range within which quadrats for a chosen habitat were located :a) Altitudinal limits- Lowland (coast to +/- 300 - 350m)b) Area- Spread over whole islandc) Distance- i) 500m min. distance from the perimeter of the Base/logistic zone Viz. none in the logistic zone.- ii) 100m min. distance from an established hut- iii) 50m min. distance from an established pathd) Aspect- East and west coasts2.Typesa) Homogeneous areasb) Least impacted areas (viz. Avoided heavily grazed Rabbit areas) (viz. Avoided Alien dominated areas) (viz. Avoided previously sampled or long term study sites)C.GENERAL SAMPLING STRATEGY FOR EACH HABITAT1.For each habitat Five 2m x 2m quadrats were located (similar in vegetation structure) and marked 1-5.2.From each quadrat two random samples were taken with the O'Connor split corer (as per sampling protocol D below). Viz: 10 cores from each habitat.3.Each sample was retained separately (in it's core-tube placed in a plastic bag) and marked accordingly. Viz: A and B from 1 through to 5 (e.g.: Poa1A-B, Poa2A-B, etc to Poa5A-B).4.On return from the field samples were immediately stored the in a cool, safe (rodent free) place (lab refrigerator) for processing.5.Invertebrate extraction followed as per protocol E below. Sample numbers were retained throughout the sampling period, together with sampling date.6.Each habitat was sampled on an average of once every five - six weeks.D.SAMPLING METHOD1.Random numbers were obtained using a table of random numbers.2.Numbers 1-100 are in top left quarter, progressing clockwise in the remaining three quarters for 101-200, 201-300 and 301-400.3.If the position chosen for the first core had already been cored, the next random number and so on was used.4.The core sample comprised a 70mm depth from ground level (viz. not including above ground vegetation growth and flowering parts).5.Care was taken to disturb as little as possible of the vegetation in and around quadrat, as well as approach to site.6.Sampling in or directly after heavy rain was avoided to prevent poor results (although it never rained hard or long enough for this situation to have occurred).7.Samples were processed within 4 days (max) after return or safe / cool storage.8.Before re-using any equipment (corer, cores, plastic bags, collecting jars and mesh cover etc), it was cleaned thoroughly to avoid contamination.E.EXTRACTION AND SORTINGMESO-INVERTEBRATES : (These include all collembola and mites and enchytraeid earthworms).1.In the collecting bottle of each sample placed in the HG extractor, an amount (+/- 2 cms high) of propylene * glycol was poured (*propylene glycol; CH3 CH(OH) CH2 OH = 76.10).2.Core-samples were separated into litter-like top and about 5- 7 cm of soil.3.Samples were retained in their respective core-rings, and where above ground vegetation biomass was more than could fit the depth of a ring, this was placed into additional rings. The veg (top)-side was covered with mesh or mutton cloth (approx. 1.5-2mm diam.) and secured with elastic bands (shock cord 3mm diam.).4.The mesh covered side was placed facing down over the collection bottle in the HG extractor. The HG was left running for the first 2 days at 25 degrees C, and for the following two days (3rd and 4th days) at 30 degrees C. 5.Samples were transferred to 99% or 100% alcohol by draining off the propylene glycol through a 60 micron mesh, picking all the colembola and mites off it with a very fine paint-brush through the view of a good microscope, and placing these into labeled vials.6.The filtered propylene glycol was re-used a couple of times.7.Where time allowed, mites and colembola were separated for certain samples.8.Sample details were noted in pencil on labels provided on the outside of each vial, and printed labels were inserted into each sample vial (see Macca Colembola and Mite labels 2001-02.doc).F.DATA ACQUISITION AND ARCHIVAL1.Field data were captured in pencil using one A6 hard-cover note-book.2.Data was transferred to spreadsheet and document and stored on CD-R discs with a back-up copy.This work was completed as part of the RiSCC project (Regional Sensitivity to Climate Change).The fields in this dataset are:Site nameHabitatLocationLatitudeLongitude

麦夸里岛微型无脊椎动物采样规程 2001年10月1日—2002年2月28日

A. 采样生境
选择了代表以下8种植被类型的典型生境：
1. 麦夸里漆姑草（Azorella macquariensis）——开阔垫状区域
2. 瘦骨木属（Acaena，包含大果瘦骨木与小瘦骨木）草本群落
3. 漆姑草状歧繁缕（Colobanthus muscoides，沿海垫状植物）
4. 高地沼泽
5. 胡克氏侧花苣苔（Pleurophyllum hookerii）优势群落区域
6. 多叶早熟禾（Poa foliosa）高大草丛
7. 短生草原（包含麦哲伦剪股颖/硬羊茅/地杨梅类）
8. 南极匙叶草（Stilbocarpa polaris）优势沿海草本群落

B. 生境点位
1. 选定生境的样方布设范围：
a) 海拔区间——低地（海岸至±300-350米）
b) 分布范围——覆盖全岛全域
c) 距离限制：
i) 与基地/后勤区边界的最小距离为500米，后勤区内不布设样方
ii) 与现有小屋的最小距离为100米
iii) 与现有步道的最小距离为50米
d) 坡向——东、西海岸区域
2. 生境类型：
a) 均质化区域
b) 受干扰程度最低的区域（即避开重度放牧兔害区域、外来物种占优区域、既往采样点或长期监测样地）

C. 各生境通用采样策略
1. 每个生境布设5个2m×2m的样方（植被结构相似），编号为1至5。
2. 从每个样方中采用奥康纳分体式采样器（O'Connor split corer，详见下述规程D）随机采集2份样本，即每个生境共采集10份取芯样本。
3. 每份样本单独保存：将取芯管放入塑料袋并做好对应标记，例如每个样方的两份样本分别标记为A、B，如Poa1A、Poa1B，依此类推至Poa5A、Poa5B。
4. 野外采样完成后，样本立即储存在阴凉、防啮齿动物的安全环境（实验室冰箱）中以待后续处理。
5. 无脊椎动物提取参照下述规程E，采样全程保留样本编号与采样日期。
6. 每个生境平均每5至6周采样一次。

D. 采样方法
1. 采用随机数表生成随机采样点位坐标。
2. 随机数1-100分布于左上象限，101-200、201-300、301-400依次按顺时针方向分布于其余三个象限。
3. 若选定的首个取芯点位已被采样，则更换为下一个随机数，以此类推。
4. 取芯样本采集地面以下70mm深度的样品，不包含地上植被的生长与开花部分。
5. 尽量减少对样方内外植被的扰动，同时规范样点接近方式。
6. 避免在大雨期间或雨后立即采样，以防样本质量不佳（尽管本次研究中未出现降雨强度大且持续时间长的情况）。
7. 样本需在返回营地或完成阴凉安全储存后的4天内完成处理，最长时限不超过4天。
8. 重复使用设备（采样器、取芯管、塑料袋、收集罐、网罩等）前，需彻底清洁以避免交叉污染。

E. 提取与分选
中型无脊椎动物（包含所有弹尾虫（Collembola）、螨类（mites）及线蚓类蚯蚓（enchytraeid earthworms））：
1. 在每个样本的收集瓶中加入约2cm高的丙二醇（propylene glycol；化学式：CH₃CH(OH)CH₂OH，摩尔质量76.10）。
2. 将取芯样本分为枯落物上层及约5-7cm深的土壤两层。
3. 样本保留在原取芯环中，若地上植被生物量超出取芯环容纳深度，则将多余部分放入额外取芯环。将植被上层一侧用网布或粗棉布（孔径约1.5-2mm）覆盖，并用直径3mm的弹力橡皮筋固定。
4. 将覆盖网布的一侧朝下放置于HG提取器的收集瓶上方，HG提取器先以25℃运行前2天，后续2天（第3、4天）以30℃运行。
5. 提取完成后，通过60μm网筛滤出丙二醇，用细画笔在高倍显微镜下挑取所有弹尾虫与螨类，转移至标注好的样品瓶中，并用99%或100%乙醇保存。
6. 过滤后的丙二醇可重复使用数次。
7. 时间允许的情况下，对部分样本的螨类与弹尾虫进行进一步分类。
8. 用铅笔将样本信息填写在样品瓶外的标签上，并将打印好的标签放入样品瓶内（详见《Macca Colembola and Mite labels 2001-02.doc》）。

F. 数据获取与归档
1. 野外数据采用A6硬皮笔记本以铅笔记录。
2. 数据转录至电子表格与文档后，存储于CD-R光盘并制作备份副本。
本研究为RiSCC项目（区域气候变化敏感性研究，Regional Sensitivity to Climate Change）的一部分。
本数据集包含以下字段：
样点名称、生境类型、地理位置、纬度、经度