粤东大规模增养殖区柘林湾海洋微生物的生态学研究
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摘要
粤东柘林湾是广东省大规模海产增养殖区之一。近年来，海水增养殖业的迅猛发展导致了该湾富营养化的加剧和生态条件的恶化，有害赤潮频发，养殖难度加大。为了了解生态退化和有害赤潮频发的原因，作者及其研究小组从2000年起对柘林湾生态系统的结构及其功能进行了长期的综合性调查。本文着重介绍有关大量营养盐分布和海洋微生物生态学的调查结果。
调查时间为2000年5月至2005年8月。其中2004和2005年的调查频率为每季度一次，其余年份则为每月一次。长期调查的站位共有9个，包括湾内7个和湾外2个。采样时用HQM-1型有机玻璃采水器采集表(离水面0.5m)、底(离水底0.5m)层水样各5L，将其中100ml水样装于无菌聚乙烯塑料瓶，冷藏带回实验室处理。沉积物样品采集使用彼得森采泥器(曙光HNM_(1-2)型，采样面积0.1m~2)，每一站点每次采样一次，取采泥器中央表层0-3cm未受干扰的泥样约50g，封装于无菌培养皿中，冷藏带回实验室进行处理。取部分水样进行菌群活体培养，其余加入经0.22μm微孔滤膜过滤的无颗粒甲醛固定，并使其最终浓度为5％(v／v)，用于测定细菌总数。将带回的沉积物样混匀后准确称取约10g，置于盛有90mL含1％Tween80无菌无颗粒海水(经0.2μm滤膜过滤)的150mL锥形瓶中，充分振荡，使微生物分散均匀后，用无菌取样器定量吸取上清液，测定方法同水样。总细菌计数采用荧光显微镜直接计数法(AODC)，异养细菌分离采用Zobell 2216E培养基平板计数法，弧菌分离采用选择性培养基TCBS平板计数法，硫酸盐还原菌(SRB)采用MPN法计数，选用乳酸钠液体培养基。氨氮(NH_4-N)、硝态氮(NO_3-N)、亚硝态氮(NO_2-N)、总无机氮(DIN)、无机磷(DIP)、硅酸盐(SiO_4-Si)等因子使用荷兰SKALAR水质微量流动注射分析仪和日本岛津UV-501紫外／可见分光光度计进行测定。温度(T)、盐度(S)、溶氧(DO)、pH等物理指标使用美国YSI6600水质分析仪现场测定。年度划分均以当年的1-12月为一个年度。数据处理与相关性分析采用软件Origin7.0进行。
调查海域溶解性无机氮、磷、硅的含量都明显偏高，总平均值分别达到31.02、1.35、36.30umol／L，其中氮、磷含量均超过国家三类海水的水质标准。由于湾顶黄冈河和湾周边排污排废的影响，营养盐的分布基本上表现出湾内高于湾外、近岸向离岸递减的格局。大规模增养殖业造成的二次污染对该湾营养盐的时空分布具有重要的影响。但以Justic和Dortch等的标准来衡量，该湾浮游植物生长受控于单一营养盐限制因子的出现率为磷46.54％，硅15.04％和氮11.46％。过量的N及高N／P值特性且持续升高的趋势可能是柘林湾地
Abstract: Zhelin Bay is one of the most important bays for large-scale mariculture in Guangdong Province, China. Due to the increasing human population and mariculture in the last decades, the ecological environment has changed dramatically, e.g., the large-scale of harmful algal bloom (Phaeocystis) occurred in 1997 and 1999, respectively. Although degenerative and eutrophic environment has brought serious economic loss and made mariculture becoming more and more difficult, especially in caged fish culture, few studies have been performed. To evaluate the environmental condition of Zhelin Bay, from May 2000 our group initiated an ecological investigation around Zhelin Bay. The paper describes the investigated results on nutrients and marine microbial ecology.
Samples were monthly collected once per month from May of 2000 to December of 2003, but only once per quarter in 2000 and 2005. There were totally 9 sampling stations, including 7 inside the bay and 2 outside the bay. Five liters of water samples were collected with HQM-1 sampling bottles 0.5 m under the surface and 0.5 m above the bottom at each station, and 100ml water from each sample were aliquoted into sterile polythylene bottles, then transported in ice to the laboratory. Samples of sediment were once collected by HNM_(1-2) grab at each station, then 50g were aliquoted from the central surface of sediment without turbulence into sterile dish and transported to laboratory for analyzing. Some water samples were used for assessing the live microorganisms, others were immediately fixed with 5% formaldehyde (final concentration) for direct count of bacteria. After 10g samples of sediment and 90ml sterile sea water (adding 1% Tween 80) filtered through 0.22um polycarbonate filters were mixed completely, supernate were for further analyze. Total bacteria were counted with AODC, heterotrophic and vibrio bacteria were measured with agar plate of Zobell 2216E and TCBS, individually. SRB were enumerated with MPN in liquid medium of sodium lactate. The NH_4-N, NO_3-N, NO_2-N, DIN and PO_4-P were analyzed by the SKALAR Channel Seawater Segmented Flow Nutrient Analyzer. TH SiO_3-Si was analyzed by the SHMADZU UV-2501 PC. Temperature, salinity, dissolved oxygen and pH were determined in situ with a portable instrument for water quality analysis (YSI6600-02, USA).Yearly division was from January to December in a same year. Relations between nutrients, the microorganism abundance and environmental variables were analyzed by means of the linear fit using the software of Origin 7.0
The results showed that the concentration of dissolved inorganic nitrogen, dissolved inorganic phosphate and silicate were high greatly, with an average concentration of 33.19、 1.48、
调查时间为2000年5月至2005年8月。其中2004和2005年的调查频率为每季度一次，其余年份则为每月一次。长期调查的站位共有9个，包括湾内7个和湾外2个。采样时用HQM-1型有机玻璃采水器采集表(离水面0.5m)、底(离水底0.5m)层水样各5L，将其中100ml水样装于无菌聚乙烯塑料瓶，冷藏带回实验室处理。沉积物样品采集使用彼得森采泥器(曙光HNM_(1-2)型，采样面积0.1m~2)，每一站点每次采样一次，取采泥器中央表层0-3cm未受干扰的泥样约50g，封装于无菌培养皿中，冷藏带回实验室进行处理。取部分水样进行菌群活体培养，其余加入经0.22μm微孔滤膜过滤的无颗粒甲醛固定，并使其最终浓度为5％(v／v)，用于测定细菌总数。将带回的沉积物样混匀后准确称取约10g，置于盛有90mL含1％Tween80无菌无颗粒海水(经0.2μm滤膜过滤)的150mL锥形瓶中，充分振荡，使微生物分散均匀后，用无菌取样器定量吸取上清液，测定方法同水样。总细菌计数采用荧光显微镜直接计数法(AODC)，异养细菌分离采用Zobell 2216E培养基平板计数法，弧菌分离采用选择性培养基TCBS平板计数法，硫酸盐还原菌(SRB)采用MPN法计数，选用乳酸钠液体培养基。氨氮(NH_4-N)、硝态氮(NO_3-N)、亚硝态氮(NO_2-N)、总无机氮(DIN)、无机磷(DIP)、硅酸盐(SiO_4-Si)等因子使用荷兰SKALAR水质微量流动注射分析仪和日本岛津UV-501紫外／可见分光光度计进行测定。温度(T)、盐度(S)、溶氧(DO)、pH等物理指标使用美国YSI6600水质分析仪现场测定。年度划分均以当年的1-12月为一个年度。数据处理与相关性分析采用软件Origin7.0进行。
调查海域溶解性无机氮、磷、硅的含量都明显偏高，总平均值分别达到31.02、1.35、36.30umol／L，其中氮、磷含量均超过国家三类海水的水质标准。由于湾顶黄冈河和湾周边排污排废的影响，营养盐的分布基本上表现出湾内高于湾外、近岸向离岸递减的格局。大规模增养殖业造成的二次污染对该湾营养盐的时空分布具有重要的影响。但以Justic和Dortch等的标准来衡量，该湾浮游植物生长受控于单一营养盐限制因子的出现率为磷46.54％，硅15.04％和氮11.46％。过量的N及高N／P值特性且持续升高的趋势可能是柘林湾地
Abstract: Zhelin Bay is one of the most important bays for large-scale mariculture in Guangdong Province, China. Due to the increasing human population and mariculture in the last decades, the ecological environment has changed dramatically, e.g., the large-scale of harmful algal bloom (Phaeocystis) occurred in 1997 and 1999, respectively. Although degenerative and eutrophic environment has brought serious economic loss and made mariculture becoming more and more difficult, especially in caged fish culture, few studies have been performed. To evaluate the environmental condition of Zhelin Bay, from May 2000 our group initiated an ecological investigation around Zhelin Bay. The paper describes the investigated results on nutrients and marine microbial ecology.
Samples were monthly collected once per month from May of 2000 to December of 2003, but only once per quarter in 2000 and 2005. There were totally 9 sampling stations, including 7 inside the bay and 2 outside the bay. Five liters of water samples were collected with HQM-1 sampling bottles 0.5 m under the surface and 0.5 m above the bottom at each station, and 100ml water from each sample were aliquoted into sterile polythylene bottles, then transported in ice to the laboratory. Samples of sediment were once collected by HNM_(1-2) grab at each station, then 50g were aliquoted from the central surface of sediment without turbulence into sterile dish and transported to laboratory for analyzing. Some water samples were used for assessing the live microorganisms, others were immediately fixed with 5% formaldehyde (final concentration) for direct count of bacteria. After 10g samples of sediment and 90ml sterile sea water (adding 1% Tween 80) filtered through 0.22um polycarbonate filters were mixed completely, supernate were for further analyze. Total bacteria were counted with AODC, heterotrophic and vibrio bacteria were measured with agar plate of Zobell 2216E and TCBS, individually. SRB were enumerated with MPN in liquid medium of sodium lactate. The NH_4-N, NO_3-N, NO_2-N, DIN and PO_4-P were analyzed by the SKALAR Channel Seawater Segmented Flow Nutrient Analyzer. TH SiO_3-Si was analyzed by the SHMADZU UV-2501 PC. Temperature, salinity, dissolved oxygen and pH were determined in situ with a portable instrument for water quality analysis (YSI6600-02, USA).Yearly division was from January to December in a same year. Relations between nutrients, the microorganism abundance and environmental variables were analyzed by means of the linear fit using the software of Origin 7.0
The results showed that the concentration of dissolved inorganic nitrogen, dissolved inorganic phosphate and silicate were high greatly, with an average concentration of 33.19、 1.48、
引文
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