CN113994946A - 邻羟基肉桂酸在清除有害底栖硅藻中的应用 - Google Patents
邻羟基肉桂酸在清除有害底栖硅藻中的应用 Download PDFInfo
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- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
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- A01N37/36—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids containing at least one carboxylic group or a thio analogue, or a derivative thereof, and a singly bound oxygen or sulfur atom attached to the same carbon skeleton, this oxygen or sulfur atom not being a member of a carboxylic group or of a thio analogue, or of a derivative thereof, e.g. hydroxy-carboxylic acids
- A01N37/38—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids containing at least one carboxylic group or a thio analogue, or a derivative thereof, and a singly bound oxygen or sulfur atom attached to the same carbon skeleton, this oxygen or sulfur atom not being a member of a carboxylic group or of a thio analogue, or of a derivative thereof, e.g. hydroxy-carboxylic acids having at least one oxygen or sulfur atom attached to an aromatic ring system
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- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
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Abstract
本发明属于海藻养殖技术领域，具体涉及邻羟基肉桂酸在清除有害底栖硅藻中的应用，本发明以邻羟基肉桂酸作为杀藻剂来清除有害底栖硅藻，由于本发明使用的杀藻剂‑邻羟基肉桂酸是异枝江蓠的提取物，与传统的清除硅藻方法相比具有以下显著优点：(1)在自然环境中易于降解，不会产生二次污染；(2)见效快，在3mM/L的邻羟基肉桂酸处理下，2天后对吸附型底栖硅藻的抑制率已超过50％，且操作简单，节约人力；(2)价格低廉，容易获取。可见，本发明的清除有害底栖硅藻方法是一种环保、高效、经济的底栖硅藻防除法。
Description
技术领域
本发明属于海藻养殖技术领域，具体涉及邻羟基肉桂酸在清除有害底栖硅藻中的应用。
背景技术
硅藻是一类具有硅质外壳的单细胞微藻，种类多样，共计多达250属10万种，并且能够通过多个个体连结成各式各样的群体。硅藻的分布范围极广，从淡水到海水区域均有大量分布。在海洋环境中，硅藻作为最重要的初级生产者，能在全球范围内构成20％的初级生产力，也是全球硅循环、碳循环的重要参与者。根据栖息状态的不同，可将硅藻分为浮游硅藻和底栖硅藻两种，浮游硅藻营漂浮生活，底栖硅藻则营固着生活。其中，底栖硅藻是构成海洋生物污损最基础的部分，浸入海水的物体表面会在几分钟内形成一层有机膜，底栖硅藻和细菌会迅速附着在这层有机膜上形成微生物膜，这种微生物膜能够给大型污损生物提供适宜的生存环境，从而形成一个复杂的垂直分布的生物群落系统。
有害底栖硅藻附着的危害主要表现在两个方面：一是对人工设施设备造成严重破坏，二是直接危害大型经济海藻产业。其中，对人工设施设备造成严重破坏的具体表现为：(1)大大提高了能耗，例如：大型污损生物固着在船体表面会造成航行阻力增加；(2)加快腐蚀，例如：污损生物的次生代谢物会加快设备表面的腐蚀速度；(3)提高维护成本，例如：船舶出航前的维护成本和时间大大增加。直接危害大型经济海藻产业的具体表现为：(1)造成海藻减产，例如：附着在礁膜、紫菜上的底栖硅藻会直接影响宿主的光合作用或者通过分泌有毒代谢物影响宿主的正常生长；(2)引起其他海藻疾病，例如：硅藻附着症会并发其他病害，从而进一步威胁海藻养殖；(3)影响海藻品质，例如：底栖硅藻附着严重的紫菜、海带上会有鸽粪大的斑点，从而降低商品的价值。
传统的有害底栖硅藻清除方法主要有物理清除法和化学清除法两种，其中，物理清除是通过人为施加的外力来提高水流速度、超声波等，从而达到清除有害底栖硅藻的目的；化学清除则是通过向底栖硅藻生长的环境中加入有毒化学物质(如硫酸铜、次氯酸钠溶液等)，从而抑制底栖硅藻的附着甚至是将其杀死。物理清除法需要耗费大量的人力和时间，需要的成本也较高，而且不能从根本上清除硅藻。化学清除法虽然效果显著，但容易造成严重的二次污染，对环境破坏极大。
因此，为有效控制底栖硅藻对大型经济海藻产业的危害，有必要探索出一种经济、低毒害、高效环保的底栖硅藻防除方法。
发明内容
为了克服上述现有技术的不足，本发明提出了邻羟基肉桂酸在清除有害底栖硅藻中的应用，即以邻羟基肉桂酸作为杀藻剂来清除有害底栖硅藻，既经济、低毒害、又高效、经济。
为了实现上述目的，本发明所采用的技术方案是：
本发明提供了邻羟基肉桂酸在清除有害底栖硅藻中的应用。
邻羟基肉桂酸，也称2-羟基肉桂酸，邻香豆酸，其CAS号为583-17-5，其结构式如下所示：
优选地，所述有害底栖硅藻包括新月菱形藻。
本发明还提供了一种清除有害底栖硅藻的方法，具体为：以邻羟基肉桂酸作为杀藻剂来清除有害底栖硅藻。
优选地，上述的一种清除有害底栖硅藻的方法，具体为：往有害底栖硅藻的生长水域中添加一定浓度的邻羟基肉桂酸作为杀藻剂，从而达到清除有害底栖硅藻的目的。
优选地，所述邻羟基肉桂酸的终浓度为0.5-3mM/L。
进一步地，所述邻羟基肉桂酸的终浓度为3mM/L。
优选地，所述有害底栖硅藻包括新月菱形藻。
优选地，所述有害底栖硅藻的生长水域为海藻养殖的海水区域。
优选地，所述有害底栖硅藻的生长水域的温度为22±1℃，盐度为30±1‰，pH为7.0-8.5。
本发明还提供了上述的一种清除有害底栖硅藻的方法在海藻养殖中的应用。
与现有技术相比，本发明的有益效果是：
本发明提供了邻羟基肉桂酸在清除有害底栖硅藻中的应用，即以邻羟基肉桂酸作为杀藻剂来清除有害底栖硅藻，由于本发明使用的杀藻剂-邻羟基肉桂酸是异枝江蓠的提取物，与传统的清除硅藻方法相比具有以下显著优点：(1)在自然环境中易于降解，不会产生二次污染；(2)见效快，在3mM/L的邻羟基肉桂酸处理下，2天后对吸附型底栖硅藻的抑制率已超过50％，且操作简单，节约人力；(2)价格低廉，容易获取。可见，本发明的清除有害底栖硅藻方法是一种环保、高效、经济的底栖硅藻防除法。
附图说明
图1为邻羟基肉桂酸处理后新月菱形藻的生长曲线；
图2为邻羟基肉桂酸对新月菱形藻的抑制效果。
具体实施方式
下面对本发明的具体实施方式作进一步说明。在此需要说明的是，对于这些实施方式的说明用于帮助理解本发明，但并不构成对本发明的限定。此外，下面所描述的本发明各个实施方式中所涉及的技术特征只要彼此之间未构成冲突就可以相互组合。
下述实施例中的实验方法，如无特殊说明，均为常规方法，下述实施例中所用的试验材料，如无特殊说明，均为可通过常规的商业途径购买得到。
实施例1邻羟基肉桂酸的获取
邻羟基肉桂酸提取自异枝江蓠。异枝江蓠采自海南省文昌市冯家湾(19°24′49″N,110°42′53″E)。取20mg异枝江蓠干粉样本于1.5mL离心管中，加入400μL提取液(乙腈：甲醇＝1：1，体积比)，涡旋混匀30s后，低温超声提取30min(5℃，40KHz)，将样品置于-20℃静置30min，4℃、13000g离心15min，移取上清液，氮气吹干,加120μL复溶液(乙腈：水＝1:1，体积比)复溶，低温超声萃取5min(5℃，40KHz)，4℃、13000g离心5min，移取上清液至带内插管的进样小瓶中上机进行质谱分析，样本制备设计6个平行。取等体积的所有样本代谢物(提取所得上清液)混合制备成质控样本(QC)，在仪器分析过程中，每8个样本中插入一个QC样本，以考察整个分析过程的重复性。检测的色谱条件为：10uL样本经BEH C18色谱柱(100mm×2.1mm i.d.,1.8μm)分离后进入质谱检测，流动相A为水(含0.1％甲酸)，流动相B为乙腈/异丙醇(1/1)(含0.1％甲酸)。分离梯度为：0-3min，流动相A从线性95％降至80％，流动相B从线性5％升至20％；3-9min，流动相A从线性80％降至5％，流动相B从线性20％升至95％；9-13min，流动相A线性维持5％，流动相B线性维持95％；13.0-13.1min，流动相A从线性5％升至95％，流动相B从线性95％降至5％；13.1-16min，流动相A线性维持95％，流动相B线性维持5％。流速为0.40mL/min，柱温为40℃。质谱条件为：样品质谱信号采集采用正负离子扫描模式，质量扫描范围(m/z)为50-1000。离子喷雾电压，正离子电压5000V，负离子电压4000V，去簇电压80V，喷雾气50psi，辅助加热气50psi，气帘气30psi，离子源加热温度500℃，20-60V循环碰撞能。上机完成之后，LC-MS原始数据导入代谢组学处理软件Progenesis QI(Waters Corporation，Milford,USA)，并采用该软件进行代谢物注释、数据预处理等，然后根据软件分析得出的结果，从上海毕得医药科技有限公司购买相应的邻羟基肉桂酸标准品(即通过线性分离检测和代谢组学软件分析得出邻羟基肉桂酸为异枝江蓠提取上清液的一种化学成分，再基于软件注释出的CAS号(583-17-5)购买相应的邻羟基肉桂酸标准品)。
实施例2一种清除吸附型底栖硅藻(以新月菱形藻为模型)的方法
(1)选择水容量为600L的PE塑料材质方形无盖水箱，箱壁经刷洗后，用浓度为5mg/L的高锰酸钾溶液反复泼洒箱壁，浸泡10分钟，再用消毒过的海水冲洗干净。
(2)往洗干净的水箱中注入新鲜的砂滤海水，进水口套一个500目的滤袋，防止敌害生物的进入，进水至500L。用50mL 30％有效氯的漂白液消毒海水，静置6小时。海水使用前，再添加9.24g硫代硫酸钠进行中和，并充分曝气3小时。
(3)往蓄有消毒海水的水箱内添加经0.1％二甲基亚砜溶解的邻羟基肉桂酸，使水箱内的邻羟基肉桂酸终浓度分别为0.5、1、3mM/L，并设置只添加0.1％二甲基亚砜的对照组。选择处于相同指数生长期的新月菱形藻细胞(购自上海光语生物科技有限公司)进行接种，接种后水箱内的新月菱形藻细胞的浓度均为175×107个/L，然后加入0.8g/L的礁膜藻体(采自湛江市硇洲岛谭北湾)，最后往水箱内加入0.5LNaNO3母液、0.5LNaH2PO4·H2O母液、1.5L Na2SiO3·9H2O母液、0.5L微量金属母液对海藻进行加富培养，让其更快更好的生长，其中NaNO3母液的浓度为8.82×10-4M，NaH2PO4·H2O母液的浓度为3.62×10-5M，Na2SiO3·9H2O母液的浓度为3.18x 10-4M,微量金属母液包括FeCl3·6H2O(1.17×10-5M)、Na2EDTA·2H2O(1.17×10-5M)、CuSO4·5H2O(3.93×10-8M)、Na2MoO4·2H2O(2.60×10-8M)、ZnSO4·7H2O(7.65×10-8M)、CoCl2·6H2O(4.20×10-8M)、MnCl2·4H2O(9.10×10-7M)，上述母液均用去离子水配制。培养过程中通过气石充空气，水温通过温控系统(智能温控潜水型电热管，属于紫铜恒温加热器，购自兴化市文鑫电热电器有限公司)维持在22±1℃，盐度维持在30±1‰，pH用碳酸氢钠维持在7.0-8.5，光照强度维持在2500±100lx，光暗周期为12h：12h，实验周期为10天，每两天对各组的新月菱形藻进行细胞计数。
通过图1、图2所示的细胞密度与抑制率统计结果可知，终浓度分别为0.5、1、3mM/L的邻羟基肉桂酸均能有效降低新月菱形藻的生长密度，且降低效果呈浓度依赖性。其中，终浓度为3mM/L的邻羟基肉桂酸试验组对新月菱形藻生长的降低效果最为显著，2天后对吸附型底栖硅藻的抑制率已超过50％，第十天时对新月菱形藻的抑制率高达93.35％。进一步观察邻羟基肉桂酸对礁膜的影响发现，处理组的礁膜藻体色泽和柔软度无明显变化，可见，0.5-3mM/L的邻羟基肉桂酸可以有效防除新月菱形藻，且对经济海藻(比如礁膜藻)的正常生长无不良影响，说明在经济海藻人工育苗和海区养殖过程中，可以通过邻羟基肉桂酸对新月菱形藻等吸附型底栖硅藻进行清除，从而提高经济海藻人工养殖的经济效益。
以上对本发明的实施方式作了详细说明，但本发明不限于所描述的实施方式。对于本领域的技术人员而言，在不脱离本发明原理和精神的情况下，对这些实施方式进行多种变化、修改、替换和变型，仍落入本发明的保护范围内。
Claims (10)
1.邻羟基肉桂酸在清除有害底栖硅藻中的应用。
2.根据权利要求1所述的应用，其特征在于，所述有害底栖硅藻包括新月菱形藻。
3.一种清除有害底栖硅藻的方法，其特征在于，以邻羟基肉桂酸作为杀藻剂来清除有害底栖硅藻。
4.根据权利要求3所述的一种清除有害底栖硅藻的方法，其特征在于，往有害底栖硅藻的生长水域中添加一定浓度的邻羟基肉桂酸作为杀藻剂，从而达到清除有害底栖硅藻的目的。
5.根据权利要求3或4所述的一种清除有害底栖硅藻的方法，其特征在于，所述邻羟基肉桂酸的终浓度为0.5-3mM/L。
6.根据权利要求5所述的一种清除有害底栖硅藻的方法，其特征在于，所述邻羟基肉桂酸的终浓度为3mM/L。
7.根据权利要求3或4所述的一种清除有害底栖硅藻的方法，其特征在于，所述有害底栖硅藻包括新月菱形藻。
8.根据权利要求4所述的一种清除有害底栖硅藻的方法，其特征在于，所述有害底栖硅藻的生长水域为海藻养殖的海水区域。
9.根据权利要求4所述的一种清除有害底栖硅藻的方法，其特征在于，所述有害底栖硅藻的生长水域的温度为22±1℃，盐度为30±1‰，pH为7.0-8.5。
10.权利要求3-9任一项所述的一种清除有害底栖硅藻的方法在海藻养殖中的应用。
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