CN102125066B - 一种缓释型复合生物除藻剂的制备方法 - Google Patents
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Abstract
本发明公开了一种缓释型复合生物除藻剂的制备方法,包括以下工序:将粒度小于40目的稻草秸秆加入到食用油与水的混合溶液中,于4-10℃,浸提5-8天,离心,过滤、灭菌,得到浸提液;食用油与水的混合溶液与稻草秸秆的体积/质量比为1000mL∶50-100g、食用油与水的体积比分别为1∶1-2.5。本发明与现有技术相比,抑藻后水面无漂浮的食用油以及死细胞,水体清澈,作用时间持久,特别适用于频繁暴发的蓝藻水华预防与治理。
Description
技术领域
本发明属于除藻剂的制备方法,尤其属于缓释复合生物除藻剂的制备方法。
背景技术
水华由藻类如蓝藻、绿藻等大量繁殖引起,其中蓝藻常常是富营养化湖泊的优势种。近年来在我国,水华频繁发生,面积逐年扩散,持续时间逐年延长。水华导致的直接后果是生态多样性减少甚至丧失,水体景观价值和服务功能降低,饮用水受到威胁,蓝藻水华的次生代谢产物微囊藻毒素(microcystins,MC)能损害肝脏,促进肝癌发生,直接威胁人类的健康和生存。
国内外在蓝藻水华的治理上,大体有物理法、化学法和生物法。但物理法大多费时、费钱、操作困难,不易大规模实施;化学法如硫酸铜、氯化铜、高锰酸钾等会造成环境的二次污染;生物学方法中的微生物防治、生物操纵技术等尚存在不少争议,如担心微生物防治的生态安全性问题,生物法中的鲢鳙鱼对蓝藻利用率较低,排出的粪便可能还会增加水体的富营养化问题等。利用植物的化感作用以及植物分泌的化感物质来进行抑藻是目前国内外广泛关注的热点。
化感作用是一种植物通过向环境中释放化学物质影响其他生物生长的现象。释放的化学物质称为化感物质。随着水体富营养化进程的加快,蓝藻水华频繁爆发,其导致的危害不断加剧,采用水生植物化感作用控制蓝藻水华发生发展越来越受到人们的重视。迄今为止,发现的化感物质主要可分为五大类:脂肪族、芳香族、含氧杂环化合物、类萜、和含氮化合物。从分离鉴定的化感物质看,脂肪酸和多酚类可能是水生植物中分布较广、含量较多的化感物质。利用水生植物的化感作用控制水华藻类生长的方式主要有3种:(1)将水生植物栽培至待处理水体,利用活体植物释放的化感物质抑制藻类;(2)将死亡干燥的植物体放入待处理水体,利用其腐败释放化感物质抑制藻类;(3)将从植物中提取的化感物质施入水体抑制藻类。
目前关于稻草秸秆的抑藻作用已有报道,但是单独稻草秸秆的水浸提液抑藻效果不好。
发明内容
本发明所要解决的技术问题是提供一种抑藻效果好的缓释型复合生物除藻剂的制备方法。
本发明解决技术问题的技术方案为:一种缓释型复合生物除藻剂的制备方法,包括以下工序:
将粒度小于40目的稻草秸秆加入到食用油与水的混合溶液中,于4-10℃,浸提5-8天,离心,过滤、灭菌,得到浸提液;食用油与水的混合溶液与稻草秸秆的体积质量比为1000mL∶50-100g、食用油与水的体积比分别为1∶1-2.5;
为防止藻细胞的反弹现象以及食用油与死亡的藻细胞漂浮的问题,
在浸提液中加入粒径小于40目筛的稻草秸秆、碳酸钙、聚乙烯醇,混合后,先预压成粗片,于40-60℃烘干至恒重,然后粉碎,制成颗粒,再过10-14目筛,整粒即可,浸提液、稻草秸秆、碳酸钙、聚乙烯醇的重量比为100∶5-10∶10-20∶1-2。
所述的食用油为金龙鱼1∶1∶1食用油。
本实验设计的机理是首先利用浸提液中的化感物质快速抑藻,然后通过食用油及其脂肪酸等物质的助溶下使添加的稻草秸秆粉末继续释放出其化感物质,从而达到作用时间长的特点。
由于食用油无毒、用量少;稻草秸秆来源广、价廉、对环境安全无毒;而碳酸钙具有很强的吸附性,且无毒、无臭、无味、分散性好;聚乙烯醇是一种白色、无毒、用途相当广泛的水溶性高分子,有较好的化学稳定性及良好的造膜性,用此法制备的颗粒不仅使得其中的食用油以及稻草中的化感物质得以缓慢释放,而且能够使悬浮于水面的油以及死细胞甚至死亡藻细胞释放的毒素、产生的异味等一起吸附沉淀。
本发明与现有技术相比,抑藻后水面无漂浮的食用油以及死细胞,水体清澈,作用时间持久,特别适用于频繁暴发的蓝藻水华预防与治理。
附图说明
图1为油水比10∶0稻草浸提液对铜绿微囊藻生长的影响
图2为油水比5∶5稻草浸提液对铜绿微囊藻生长的影响
图3为油水比4∶6稻草浸提液对铜绿微囊藻生长的影响
图4为油水比3∶7稻草浸提液对铜绿微囊藻生长的影响
图5为油水比0∶10稻草浸提液对铜绿微囊藻生长的影响
图6为缓释型复合抑藻剂对铜绿微囊藻生长的影响
图7为缓释型复合抑藻剂对水华鱼腥藻生长的影响
图8为缓释型复合抑藻剂对斜生栅藻生长的影响
图9为缓释型复合抑藻剂对蛋白核小球藻生长的影响
图10为缓释型复合抑藻剂对巢湖水体混合藻类生长的影响。
具体实施方式
下面结合实施例对本发明作详细的说明。
实施例1:
a、制备:
取金龙鱼油(1∶1∶1)与水的混合液1000ml(油与水的体积比分别为10∶0、5∶5、4∶6、3∶7、2∶8、1∶9、0∶10),浸提50g稻草秸秆粉末(粉末过40目筛),4℃浸提7天后,离心、过滤,灭菌,得到浸提液。
b、浸提液的抑藻作用
取浸提液按0、0.5、1.0、2.0、4.0、8.0ml/L添加至初始浓度为3.3×105个藻细胞/mL的铜绿微囊藻藻液中进行抑藻,共实验8天,每隔一天进行藻细胞计数,计算其抑制率。
抑制率的计算公式为:IR(%)=(1-N/N0)×100,
其中:IR-抑制率;N-实验组藻密度(个/ml);N0-对照组藻密度(个/ml)。
将抑藻剂浓度与抑制率作一元线性回归,求出半效应浓度EC50。
结果如图1-5所示,实验表明油水混合的稻草秸秆浸提液其抑藻效果依次为油水比5∶5>4∶6>3∶7>10∶0>2∶8>1∶9>0∶10,实验的第8天抑制率最高,分别为96.6%、92.6%、92.0%、71.7%、67.3%、47.6%、32.8%,油水比5∶5、4∶6、3∶7三组与稻草秸秆的水浸提液相比,具有极显著性差异,P<0.001,与10∶0、2∶8以及1∶9组相比,P<0.01,但由于油水比5∶5、4∶6、3∶7三个浓度间的抑制率没有显著性差异。考虑到成本,在制备缓释型复合抑藻剂时,选用金龙鱼食用油用量少的3∶7比例浸提稻草秸秆。
上述抑藻实验结束后,于实验的第9天向各藻液中添加相应培养基一次,继续观察藻液颜色变化,各组于第11天开始出现了部分反弹现象,表明了浸提液中化感物质的有效作用时间约为10天。另一缺点是浸提液中食用油以及死亡的藻细胞等漂浮于水面影响景观。
实施例2:
为解决反弹现象以及食用油与死亡的藻细胞漂浮等问题,在浸提液中加入粒径小于40目筛的稻草秸秆、碳酸钙、聚乙烯醇,混合后,于50℃烘干至恒重,再过10-14目筛,整粒,得到抑藻剂;浸提液、稻草秸秆、碳酸钙、聚乙烯醇的重量比为100∶5∶10∶1。
实施例3:
除
浸提液、稻草秸秆、碳酸钙、聚乙烯醇的重量比为100∶10∶20∶2;外,其余与实施例2相同。
实施例4:
将实施例2所制的抑藻剂分别按2.0、3.0、4.0、5.0、6.0mg/L添加至铜绿微囊藻藻液、水华鱼腥藻藻液、蛋白核小球藻藻液、斜生栅藻藻液(各藻液初始藻密度约为3~5×105个藻细胞/mL,铜绿微囊藻由BG-11培养基培养、水华鱼腥藻由HGZ培养基培养、蛋白核小球藻和斜生栅藻均采用HB-4培养基培养)以及巢湖水体混合藻液中(取已爆发水华的巢湖水样,取样后用100目的塑料纱网过滤以除去群体藻及小型水生动物,过滤后的藻液经鉴定主要优势种为蓝藻门(Cyanophyta)的微囊藻属和绿藻门(Chlorophyta)的小球藻(Chlorella),然后在藻液中添加BG-11培养基驯化培养48h再进行抑藻实验,混合藻液叶绿素a含量为1.2mg/L。)
各组置光照培养箱中进行培养,培养条件:光照度4000Lx,光周期12h:12h,温度(24±2)℃,每天摇动锥形瓶3~4次,共实验16天,于实验的第9天向各藻液中添加相应培养基一次以保证营养的供应。实验期间每隔一天取藻液进行藻细胞计数或测定藻细胞叶绿素a含量(巢湖水体混合藻液),共测定8次。
结果如图6-10、表1所示,
表1
抑藻剂的抑藻效果显著,第8与第16天对铜绿微囊藻抑制率分别达92.6%和95.4%,EC50分别为3.1247g和2.9443g;对巢湖水体混合藻最大抑制率达88.6%和89.3%,EC50分别为3.5149g和3.3275g,对各藻的抑制效果依次表现为铜绿微囊藻>巢湖混合藻>水华鱼腥藻>蛋白核小球藻>斜生栅藻,之所以对巢湖混合藻的作用较好,原因是因为混合藻中以铜绿微囊藻最多。继续观察后期水体颜色、透明度,20天后水体肉眼观测仍为无色,未见有反弹迹象。
所述的HB-4培养基的成分:
(NH4)2SO4 0.2g/L,K2HPO4 0.015g/L,CaCl2 0.015g/L,MgSO4·7H2O 0.08g/L,NaHCO3 0.1g/L,KCl 0.025g/L,FeCl3(1%液)0.15mL/L,土壤浸提液2mL/L,A5solution 0.1mL/L(A5溶液配方:蒸馏水1000mL,H3BO3 61.0mg,MnSO4·H2O169.0mg,ZnSO4·7H2O 287.0mg,CuSO4·5H2O 2.5mg)。
土壤浸提液的制备:取土400g,加水1000mL后搅匀,浸泡一天,隔水蒸3-4h,重复蒸2-3次。上清液过滤,灭菌,冷藏,保存。
BG-11培养基的成分:
NaNO3 1.5g/L,K2HPO4·3H2O 0.04g/L,MgSO4·7H2O 0.075g/L,CaCl2·2H2O0.036g/L,柠檬酸0.006g/L,柠檬酸铁铵0.006g/L,EDTA 0.001g/L,Na2CO30.02g/L,蒸馏水919mL/L。
HGZ培养基的成分
NaNO3 49.6mg,K2HPO4 3.9mg,MgSO4.7H2O 7.5mg,CaCl2.2H2O 3.6mg,Na2SiO3.9H2O 5.8mg,PIV溶液0.3mL(FeCl3.6H2O 9.7mg,MnCl2.4H2O 4.1mg,ZnCl20.5mg,CoCl2.6H2O 0.2mg,Na2MoO4.2H2O 0.4mg,Na2-EDTA 75mg,蒸馏水100mL。),土壤提取液0.3mL,蒸馏水99.4mL。
Claims (2)
1.一种缓释型复合生物除藻剂的制备方法,其特征在于:包括以下工序:
将粒度小于40目的稻草秸秆加入到食用油与水的混合溶液中,于4-10℃,浸提5-8天,离心,过滤、灭菌,得到浸提液;食用油与水的混合溶液与稻草秸秆的体积/质量比为1000mL:50-100g、食用油与水的体积比分别为1:1-2.5;
在浸提液中加入粒径小于40目筛的稻草秸秆、碳酸钙、聚乙烯醇,混合后,先预压成粗片,于40-60℃烘干至恒重,然后粉碎,制成颗粒,再过10-14目筛,整粒即可,浸提液、稻草秸秆、碳酸钙、聚乙烯醇的重量比为100:5-10:10-20:1-2。
2.根据权利要求1所述的一种缓释型复合生物除藻剂的制备方法,其特征在于:所述的食用油为金龙鱼1:1:1食用油。
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