技术领域technical field
本发明涉及微生物学，特别是涉及一种硅藻培养装置与方法。The invention relates to microbiology, in particular to a diatom culture device and method.
背景技术Background technique
底栖硅藻含有丰富的营养物质，是鲍鱼、海参和海胆等名贵水产动物苗种的重要饵料。与浮游微藻相比，底栖硅藻脱水采收的成本较低，但由于底栖硅藻的附着特性，单位水体的生物质产量远低于浮游微藻如小球藻的生物质产量。Benthic diatoms are rich in nutrients and are important bait for rare aquatic animal seedlings such as abalone, sea cucumber and sea urchin. Compared with planktonic microalgae, the cost of dehydration and harvesting of benthic diatoms is lower, but due to the attachment characteristics of benthic diatoms, the biomass production per unit of water body is much lower than that of planktonic microalgae such as Chlorella.
在生产实践中，底栖硅藻的培养仍普遍采用自然海区挂板附着或刮砂淘洗等传统的培养方法，在这种培养条件下，各种理化因子不稳定，生物质产量较低，质量也得不到保证，无法满足生产上的需求。In production practice, traditional culture methods such as hanging board attachment or sand scraping and elutriation in natural sea areas are still commonly used for the cultivation of benthic diatoms. Under such cultivation conditions, various physical and chemical factors are unstable, and the biomass yield is low. The quality is not guaranteed and cannot meet the needs of production.
发明内容Contents of the invention
本发明要解决的技术问题是提供一种结构简单、有效提高硅藻生物质产量的硅藻培养装置与方法。The technical problem to be solved by the present invention is to provide a diatom cultivation device and method which has a simple structure and can effectively improve the yield of diatom biomass.
本发明的硅藻培养装置，包括用于盛装培养基的培养容器以及置于培养容器内部的硅藻附板，还包括置于培养容器内部的用于向硅藻附板吹送空气的充气管，所述充气管与充气泵连接。The diatom culture device of the present invention comprises a culture container for containing medium and a diatom attachment board placed inside the culture container, and also includes an air-filled tube for blowing air to the diatom attachment board placed inside the culture container, The inflation tube is connected with an air pump.
本发明的硅藻培养装置，其中，还包括置于培养容器内部的用于支承硅藻附板的附板架，所述附板架包括底板以及分别垂直连接于底板的四个侧边上四个侧栏，四个所述侧栏依次首尾连接，所述硅藻附板垂直于所述底板，所述硅藻附板的两端分别伸入两相对所述侧栏的相对空隙中，以此所述附板架支承硅藻附板。The diatom cultivating device of the present invention, wherein, also includes the attached plate frame for supporting the diatom attached plate placed inside the culture container, the attached plate frame includes a bottom plate and four sides vertically connected to the bottom plate respectively. four side rails, the four side rails are connected end to end in sequence, the diatom attachment board is perpendicular to the bottom plate, and the two ends of the diatom attachment board extend into the relative gaps of the two opposite side rails respectively, so as to The attached board frame supports the diatom attached board.
本发明的硅藻培养装置，其中，还包括用于向硅藻附板照射光线的光源以及对培养基加热的加热棒，所述加热棒置于培养容器的内部。The diatom cultivation device of the present invention further includes a light source for irradiating light to the diatom attached plate and a heating rod for heating the culture medium, and the heating rod is placed inside the culture container.
本发明的硅藻培养装置，其中，硅藻附板包括塑料的方形框架、以及覆盖于所述方形框架上的40-100目的透明的塑料纱网。In the diatom cultivation device of the present invention, the diatom attachment plate comprises a plastic square frame and a transparent plastic gauze of 40-100 meshes covering the square frame.
本发明的硅藻培养装置，其中，所述方形框架包括面积较大的第一塑料框和面积较小的第二塑料框，第一塑料框的四边与第二塑料框的四边相配合，以将所述塑料纱网夹持固定。In the diatom culturing device of the present invention, the square frame includes a first plastic frame with a larger area and a second plastic frame with a smaller area, and the four sides of the first plastic frame are matched with the four sides of the second plastic frame, so as to The plastic gauze is clamped and fixed.
本发明的硅藻培养装置，其中，所述充气管为塑料管，所述充气管上设置由多个通气孔，相邻通气孔的距离为3-5厘米，所述充气管为多个，多个充气管缠绕于硅藻附板上，相邻的充气管的管间距为30-50厘米。The diatom culture device of the present invention, wherein, the described gas-filled tube is a plastic tube, and the described gas-filled tube is provided with a plurality of vent holes, the distance between adjacent vent holes is 3-5 cm, and the gas-filled tube is multiple, A plurality of inflatable tubes are wound on the diatom appendage, and the distance between adjacent inflatable tubes is 30-50 cm.
本发明的硅藻培养装置，其中，所述充气管为纳米管，所述充气管平铺于培养容器的底部。In the diatom culture device of the present invention, the gas-filled tube is a nanotube, and the gas-filled tube is flatly laid on the bottom of the culture container.
本发明的硅藻培养装置，其中，光源包括设置于培养容器上方的顶部光源和设置于培养容器两侧的侧面光源。In the diatom cultivation device of the present invention, the light source includes a top light source arranged above the culture container and side light sources arranged on both sides of the culture container.
本发明的硅藻培养装置，其中，还包括用于控制充气泵以及光源的计时开关，计时开关与所述充气泵以及光源的电源分别连接。The diatom cultivation device of the present invention further includes a timing switch for controlling the air pump and the light source, and the timing switch is connected to the power supply of the air pump and the light source respectively.
本发明的硅藻培养方法，包括：The diatom culture method of the present invention comprises:
藻种培养步骤；Algae cultivation step;
大量培养步骤：向培养容器中加培养基，将上步骤培养的藻种置于培养容器中，多个硅藻附板间隔3厘米垂直置于附板架上，向硅藻附板吹送空气，对硅藻附板照射灯光；A large number of cultivation steps: add culture medium to the culture container, place the algae species cultivated in the previous step in the culture container, place a plurality of diatom attached plates vertically on the attached plate frame at intervals of 3 cm, blow air to the diatom attached plates, Irradiate light on the diatom attached plate;
采收步骤：在培养容器中培养3至5天后，取出硅藻附板，放置于添加起膜剂的海水中，1至2天后，待附着于硅藻附板上的硅藻细胞呈膜状或片状悬浮在水体中，用网筛过滤采收硅藻细胞，其中，起膜剂由NH3SO4和NaNO3组成，Harvesting steps: After cultivating in the culture container for 3 to 5 days, take out the diatom attached plate and place it in seawater with a film-forming agent added. After 1 to 2 days, the diatom cells attached to the diatom attached plate will be in the form of a film Or flakes suspended in the water body, and the diatom cells are harvested by filtering with a mesh screen, wherein the film-forming agent is composed of NH3 SO4 and NaNO3 ,
或，or,
将硅藻附板投于动物养殖池中，待动物舔食干净。Throw the diatom attached board into the animal breeding pond, and wait for the animals to lick it clean.
上述的大量培养步骤中，具体的，按藻种培养体积与培养基体积比1：20-120的比例加入藻种培养液，或者收集的湿藻种In the above-mentioned large-scale cultivation steps, specifically, the algae culture solution is added according to the ratio of the algae culture volume to the medium volume ratio of 1:20-120, or the collected wet algae
所述方法中，其中，起膜剂优选由下述重量份数比的组分组成NH3SO4:NaNO3＝2-7：1，优选NH3SO4:NaNO3＝5:1，使用时，溶于F/2培养基中，配成20～80mg/L浓度的起膜剂溶液。In the method, wherein, the film-forming agent is preferably composed of the following components in parts by weight: NH3 SO4 :NaNO3 =2-7:1, preferably NH3 SO4 :NaNO3 =5:1, using , dissolve in F/2 medium, and prepare a film-forming agent solution with a concentration of 20-80 mg/L.
上述硅藻可以为底栖硅藻，如菱形藻属、双眉藻属等等，所述大量培养步骤中，培养的具体条件可以根据不同硅藻的生长条件设置，比如，在本发明的一个实施方式中，对于菱形藻Nitzschia sp.、双眉藻，Amphora sp.，大量培养步骤的条件为使海水培养基的温度为25-38℃，对硅藻附板照射灯光，光照强度10-80mol.m-2.s-1，使昼：夜比为14小时：10小时。Above-mentioned diatoms can be benthic diatoms, such as Nitzphylla, Bacteria, etc., in the large-scale cultivation step, the specific conditions of cultivation can be set according to the growth conditions of different diatoms, for example, in one of the present invention In an embodiment, for Nitzschia sp., Amphora sp., the conditions of the large-scale cultivation step are to make the temperature of the seawater culture medium be 25-38°C, irradiate the diatom attached plate with light, and the light intensity is 10-80mol .m-2 .s-1 , so that the day: night ratio is 14 hours: 10 hours.
其中，对于双眉藻属，例如Amphora coffeaeformis和Amphora exigua还包括将采收后的硅藻细胞集中起来，置于5-25℃低温环境中给予低温刺激0.5-2天，低温刺激后的藻细胞油脂含量可提高50％左右，EPA含量可提高2-5倍。Among them, for the genus Amphora coffeaeformis and Amphora exigua, for example, the diatom cells after harvesting are collected and placed in a low temperature environment of 5-25°C to give low temperature stimulation for 0.5-2 days, and the algal cells after low temperature stimulation The oil content can be increased by about 50%, and the EPA content can be increased by 2-5 times.
本发明的技术方案，利用底栖硅藻的附着的生物学特性，将细胞培养在固体培养材料上，经起膜剂作用后，附着于固体材料上的硅藻细胞会脱离附着基质，通过网筛过滤即可采收，免除了刮取附着细胞的过程，也节约了一般浮游微藻离心脱水时所需能耗。The technical scheme of the present invention utilizes the attached biological characteristics of benthic diatoms to culture the cells on the solid culture material. After acting as a film agent, the diatom cells attached to the solid material will break away from the attachment matrix and pass through the net. It can be harvested by sieving and filtering, which avoids the process of scraping off the attached cells, and also saves the energy required for the centrifugal dehydration of general planktonic microalgae.
本发明的技术方案，针对现有微藻液体养殖中条件苛刻、能耗高、易污染和细胞密度低的问题，提供一种结构简单、有效提高硅藻生物质产量的硅藻培养装置与方法，使用本发明的技术方案培养微藻不需要严苛的条件，能耗较低，不易发生污染，并且能够产生高密度的藻细胞。The technical scheme of the present invention aims at the problems of harsh conditions, high energy consumption, easy pollution and low cell density in the existing microalgae liquid cultivation, and provides a diatom cultivation device and method with a simple structure and effective improvement of diatom biomass production , using the technical solution of the present invention to cultivate microalgae does not require harsh conditions, has low energy consumption, is not prone to pollution, and can produce high-density algae cells.
附图说明Description of drawings
图1为本发明的硅藻培养装置的结构示意图的主剖视图；Fig. 1 is the main sectional view of the structural representation of diatom culture device of the present invention;
图2为本发明的硅藻培养装置的结构示意图的左视图；Fig. 2 is the left view of the structural representation of diatom cultivation device of the present invention;
图3为附板架的结构示意图的立体图；Fig. 3 is the three-dimensional view of the structural representation of attached board frame;
图4为硅藻附板的结构示意图的主视图；Fig. 4 is the front view of the structural representation of diatom attached plate;
图5为另一种实施例的硅藻附板的结构示意图的主视图。Fig. 5 is a front view of a structural schematic diagram of a diatom attached plate of another embodiment.
具体实施方式Detailed ways
如图1、图2所示，本发明的硅藻培养装置，包括用于盛装培养基的培养容器1以及置于培养容器1内部的硅藻附板2，还包括置于培养容器1内部的用于向硅藻附板2吹送空气的充气管3，充气管3与充气泵连接。As shown in Fig. 1 and Fig. 2, the diatom culture device of the present invention includes a culture container 1 for containing culture medium and a diatom attachment plate 2 placed inside the culture container 1, and also includes a culture container placed inside the culture container 1. An inflatable tube 3 for blowing air to the diatom attached plate 2, and the inflatable tube 3 is connected with an air pump.
培养容器1的容积可以为5升-500升，材质可以是玻璃或塑料或水泥。The volume of the culture container 1 can be 5 liters-500 liters, and the material can be glass, plastic or cement.
结合图3、图4所示，本发明的硅藻培养装置，其中，还包括置于培养容器1内部的用于支承硅藻附板2的附板架4，附板架4包括底板41以及分别垂直连接于底板41的四个侧边上四个侧栏42，四个侧栏42依次首尾连接，硅藻附板2垂直于底板41，硅藻附板2的两端分别伸入两相对侧栏的相对空隙中，以此附板架4支承硅藻附板2。In conjunction with Fig. 3, shown in Fig. 4, the diatom culture device of the present invention, wherein, also comprise the attached plate frame 4 that is used to support diatom attached plate 2 inside the culture container 1, attached plate frame 4 comprises bottom plate 41 and Respectively vertically connected to four side rails 42 on the four sides of the base plate 41, the four side rails 42 are connected end to end in turn, the diatom attachment plate 2 is perpendicular to the base plate 41, and the two ends of the diatom attachment plate 2 extend into two opposite sides respectively. In the relative gap of the side rail, the attached board frame 4 supports the diatom attached board 2 with this.
本发明的硅藻培养装置，其中，附板架4可放置多块硅藻附板，根据培养容器长度调节硅藻附板的数量，养容器可大可小，每3-5厘米放一硅藻附板。In the diatom cultivating device of the present invention, wherein, the attached board frame 4 can place a plurality of diatom attached boards, and the number of diatom attached boards can be adjusted according to the length of the culture container. Algae attached board.
本发明的硅藻培养装置，其中，还包括用于向硅藻附板照射光线的光源以及用于对培养基加热的加热棒，加热棒置于培养容器1的内部。The diatom culture device of the present invention further includes a light source for irradiating light to the diatom attached plate and a heating rod for heating the culture medium, and the heating rod is placed inside the culture container 1 .
本发明的硅藻培养装置，其中，硅藻附板2包括塑料的方形框架21以及覆盖于方形框架21上的40-100目的透明的塑料纱网22。In the diatom cultivation device of the present invention, the diatom attachment plate 2 includes a plastic square frame 21 and a 40-100 mesh transparent plastic gauze 22 covering the square frame 21 .
本发明的硅藻培养装置，其中，方形框架21包括面积较大的第一塑料框和面积较小的第二塑料框，第一塑料框的四边与第二塑料框的四边相配合，以将塑料纱网22夹持固定。In the diatom cultivating device of the present invention, the square frame 21 comprises a first plastic frame with a larger area and a second plastic frame with a smaller area, and the four sides of the first plastic frame cooperate with the four sides of the second plastic frame to The plastic gauze 22 is clamped and fixed.
本发明的硅藻培养装置，其中，硅藻附板2使藻细胞采收方便，节约培养空间。In the diatom cultivation device of the present invention, the diatom attachment plate 2 facilitates the harvesting of algae cells and saves cultivation space.
塑料纱网22的材料可以是聚乙烯或乙烯或尼龙。The material of plastic gauze 22 can be polyethylene or ethylene or nylon.
结合图5所示，在本发明的硅藻培养装置的其他实施例中，可以利用U形封边条8将塑料纱网22固定于方形框架21上。As shown in FIG. 5 , in other embodiments of the diatom cultivation device of the present invention, the plastic gauze 22 can be fixed on the square frame 21 by using a U-shaped edge band 8 .
本发明的硅藻培养装置，其中，充气管3为塑料管，充气管3上设置由多个通气孔6，相邻通气孔6的距离为3-5厘米，充气管3为多个，多个充气管3缠绕于硅藻附板2上，相邻的充气管3的管间距为30-50厘米。充气管3与充气泵连接，充气泵的功率、充气管的长短根据培养容器大小调整，培养容器的体积很大时，可设多组充气管。充气管用于向培养容器内通入空气或空气与二氧化碳的混和气体。In the diatom cultivation device of the present invention, wherein the gas-filled tube 3 is a plastic tube, and the gas-filled tube 3 is provided with a plurality of vent holes 6, the distance between adjacent vent holes 6 is 3-5 centimeters, and the gas-filled tube 3 is multiple, many Two inflatable tubes 3 are wound on the diatom attached plate 2, and the tube spacing of adjacent inflatable tubes 3 is 30-50 centimeters. Inflatable tube 3 is connected with air pump, and the power of air pump, the length of inflatable tube are adjusted according to the size of culture container, and when the volume of culture container is very large, multiple groups of inflatable tubes can be established. The inflatable tube is used for introducing air or a mixed gas of air and carbon dioxide into the culture container.
充气管3可使通气均匀，防止氧气积累，促进微藻的光合作用，提高生物质产量。The inflatable tube 3 can make ventilation uniform, prevent oxygen accumulation, promote photosynthesis of microalgae, and increase biomass production.
本发明的硅藻培养装置的其他实施例中，充气管可以为纳米管，充气管平铺于培养容器的底部，这样充气管还可有效防止蓝藻和绿藻等浮游微藻及浮游动物的污染。In other embodiments of the diatom culture device of the present invention, the gas-filled tube can be a nanotube, and the gas-filled tube is laid flat on the bottom of the culture container, so that the gas-filled tube can also effectively prevent the pollution of planktonic microalgae and zooplankton such as blue-green algae and green algae. .
本发明的硅藻培养装置，其中，光源包括设置于培养容器1上方的顶部光源51和设置于培养容器两侧的侧面光源52。顶部光源51和侧面光源52为荧光灯或LED。In the diatom cultivation device of the present invention, the light source includes a top light source 51 disposed above the culture container 1 and side light sources 52 disposed on both sides of the culture container. The top light source 51 and the side light source 52 are fluorescent lamps or LEDs.
顶部光源51悬挂于培养容器1上方的吊杆上。侧面光源52可以为外置和内置两种，如果培养容器1为透明的，则采用长形灯管悬挂于培养容器1的两侧，如果培养容器1是非透明容器，可将防水的LED灯条固定于培养容器1的内壁。侧面光源的设置可根据硅藻对光线的需求而定。对光照要求不高的硅藻可以不设侧面光源。The top light source 51 is suspended on the suspension rod above the culture container 1 . The side light source 52 can be two kinds of external and built-in, if the culture container 1 is transparent, then use the elongated lamp tube to hang on the both sides of the culture container 1, if the culture container 1 is a non-transparent container, you can put a waterproof LED light bar Fixed on the inner wall of the culture container 1. The setting of the side light source can be determined according to the demand of diatoms for light. Diatoms that do not have high requirements for light can not be equipped with side light sources.
本发明的硅藻培养装置，其中，还包括用于控制充气泵以及光源的计时开关，计时开关与充气泵以及光源的电源分别连接，计时开关控制充气泵以及光源的电源，在具体的实施例中，可根据微藻生物学特性设定通气和照明电源开关周期为开14小时、关10小时。The diatom cultivating device of the present invention further includes a timing switch for controlling the air pump and the light source, the timing switch is connected to the power supply of the air pump and the light source respectively, and the timing switch controls the power supply of the air pump and the light source. In a specific embodiment According to the biological characteristics of microalgae, the ventilation and lighting power switch cycle can be set as 14 hours on and 10 hours off.
本发明的硅藻培养装置，一次性投资少且运行成本低，操作简单，不必使用二氧化碳发生设备，且具有显著降低杂藻和原生动物污染等优点。本发明的硅藻培养装置培养的几种底栖硅藻干生物质产量可达到5-10g/㎡，约1g/L。The diatom cultivation device of the present invention has the advantages of less one-time investment, low operation cost, simple operation, no need to use carbon dioxide generating equipment, and significantly reduced pollution of miscellaneous algae and protozoa. The dry biomass output of several benthic diatoms cultivated by the diatom cultivation device of the present invention can reach 5-10g/㎡, about 1g/L.
本发明的硅藻培养装置实施本发明的硅藻培养方法，包括：The diatom cultivation device of the present invention implements the diatom cultivation method of the present invention, comprising:
藻种培养步骤：海水加SiO2培养基培养藻种；Algae culturing step: seawater plus SiO2 medium to cultivate algae;
大量培养步骤：向培养容器中加海水培养基，将上步骤培养的藻种按藻种培养体积与海水培养基体积比1：20—1：120置于培养容器中，多个硅藻附板间隔3厘米垂直置于附板架上，向硅藻附板吹送空气，对硅藻附板照射灯光；Large-scale cultivation step: add seawater medium to the culture container, place the algae cultured in the previous step in the culture container according to the volume ratio of algae culture volume to seawater medium volume 1:20-1:120, and attach multiple diatoms to the plate Place it vertically on the attached plate frame at an interval of 3 cm, blow air to the diatom attached plate, and irradiate the diatom attached plate with light;
采收步骤：在培养容器中培养3至5天后，取出硅藻附板，放置于添加起膜剂的海水中，1至2天后，待附着于硅藻附板上的硅藻细胞呈膜状或片状悬浮在水体中，用网筛过滤采收硅藻细胞，Harvesting steps: After culturing in the culture container for 3 to 5 days, take out the diatom attached plate and place it in seawater with film-forming agent added. After 1 to 2 days, the diatom cells attached to the diatom attached plate will be in the form of a film Or flakes suspended in the water body, the diatom cells were collected by filtering with a mesh sieve,
或，or,
将硅藻附板投于动物养殖池中，待动物舔食干净。Throw the diatom attached board into the animal breeding pond, and wait for the animals to lick it clean.
其中，起膜剂由下述重量份数比的组分组成NH3SO4:NaNO3＝2-7：1，优选NH3SO4:NaNO3＝5:1，使用时，溶于F/2培养基中，配成20～80mg/L浓度的起膜剂溶液。Among them, the film-forming agent is composed of the following components in parts by weight: NH3 SO4 :NaNO3 =2-7:1, preferably NH3 SO4 :NaNO3 =5:1, when used, dissolve in F/ 2 in the culture medium to prepare a film-forming agent solution with a concentration of 20-80 mg/L.
利用本发明的硅藻培养装置实施本发明的硅藻培养方法的实施例一Embodiment 1 of utilizing the diatom cultivation device of the present invention to implement the diatom cultivation method of the present invention
本实施例对两种底栖硅藻进行培养实验，即藻种一：菱形藻Nitzschia sp.、藻种二：双眉藻，Amphora coffeaeformisIn this example, two kinds of benthic diatoms were cultured, that is, algae species one: Nitzschia sp., algae species two: Amphora coffeeeformis
藻种培养步骤：利用F/2培养基培养藻种；30-38℃静置培养，培养至藻株浓度为0.1g/L。Algae cultivation steps: use F/2 medium to cultivate algae; culture at 30-38°C until the concentration of algae is 0.1g/L.
F/2培养基为每升海水中添加如下元素：NaNO3 75mg,NaH2PO4·2H2O5.65mg,Na2EDTA 4.16mg,FeCl3·6H2O 3.15mg,Na2SiO3·9H2O 20mg，CuSO4·5H2O 0.01mg,ZnSO4·7H2O0.022mg,CoCl2·6H2O 0.01mg,MnCl2·4H2O0.18mg,Na2MoO4·2H2O 0.006mg,Vitamin B120.0005mg,Vitamin B1 0.1mg,Biotin0.0005mg。For F/2 medium, the following elements are added per liter of seawater: NaNO3 75mg, NaH2 PO4 2H2 O 5.65mg, Na2 EDTA 4.16mg, FeCl3 6H2 O 3.15mg, Na2 SiO3 9H2 O 20mg, CuSO4 5H2 O 0.01mg, ZnSO4 7H2 O 0.022mg, CoCl2 6H2 O 0.01mg, MnCl2 4H2 O 0.18mg, Na2 MoO4 2H2 O 0.006mg, Vitamin B12 0.0005mg, Vitamin B1 0.1mg, Biotin 0.0005mg.
大量培养步骤：以50L的半透明塑料箱为培养容器，向容器中加入40L的F/2海水培养基，将100mg(湿重)藻种倒入培养基中，将40-80目聚乙烯纱网固定于塑料边框上，形成用于硅藻附着的硅藻附板，多个硅藻附板间隔3厘米垂直置于附板架上，向硅藻附板吹送空气，使海水培养基的温度为30-38℃，对硅藻附板照射灯光，光照强度10-80mol.m-2.s-1，使硅藻感受到的昼：夜比为14小时：10小时。Large-scale cultivation steps: use a 50L translucent plastic box as a culture container, add 40L of F/2 seawater medium into the container, pour 100mg (wet weight) of algae seeds into the medium, and put 40-80 mesh polyethylene yarn The net is fixed on the plastic frame to form a diatom attachment board for diatom attachment. Multiple diatom attachment boards are vertically placed on the attachment board frame at intervals of 3 cm, and air is blown to the diatom attachment board to make the temperature of the seawater culture medium At 30-38°C, light is irradiated on the diatom attached plate, and the light intensity is 10-80mol.m-2 .s-1 , so that the day:night ratio felt by the diatoms is 14 hours:10 hours.
三天后，纱网上附满黄褐色硅藻。将附满硅藻的纱网取下，放入幼参养殖池中，海参对纱网上的硅藻积极取食。将其余纱网取下，搓下纱网上附着的硅藻，通过静止分层、脱水、干燥，得微藻固体。菱形藻的干生物质产量约为5.6g/㎡，双眉藻的干生物质产量约为10.5g/㎡，利用纱网培养的微藻生物质产量是常用方法以PC板为附板的八倍。Three days later, the gauze was covered with yellow-brown diatoms. The gauze covered with diatoms is removed and put into the young ginseng breeding pond, and the sea cucumbers actively feed on the diatoms on the gauze. The rest of the gauze is removed, the diatoms attached to the gauze are rubbed off, and the solid microalgae is obtained through static layering, dehydration, and drying. The dry biomass yield of Nitzschia is about 5.6g/㎡, and the dry biomass yield of Amphibious algae is about 10.5g/㎡. The biomass yield of microalgae cultured with gauze net is a common method with PC board as the attached board. times.
在传统的微藻液体养殖中，大量的能量消耗在水体运行、搅拌、通气、二氧化碳补给和收获上。使用本发明的方法培养微藻不需要严苛的条件，设备简单，易操作，能耗较低，并且能够产生高密度的藻细胞，适合水产养殖应用。此外，实验表明，硅藻在纱网上的附着强度高于生产上常用的PC板，更利于动物取食消化。因为动物在取食附差强度高的硅藻时，硅藻壳更易破壁，有利于动物消化。In traditional microalgae liquid culture, a large amount of energy is consumed in water body operation, stirring, aeration, carbon dioxide replenishment and harvesting. Using the method of the invention to cultivate microalgae does not require harsh conditions, the equipment is simple, easy to operate, low in energy consumption, and can produce high-density algae cells, which is suitable for aquaculture applications. In addition, experiments have shown that the adhesion strength of diatoms on gauze is higher than that of PC boards commonly used in production, which is more conducive to animal feeding and digestion. Because when animals feed on diatoms with high adhesion strength, diatom shells are easier to break, which is beneficial to animal digestion.
利用本发明的硅藻培养装置实施本发明的硅藻培养方法的实施例二Utilize the diatom cultivation device of the present invention to implement the embodiment two of the diatom cultivation method of the present invention
本实施例对双眉藻属，Amphora coffeaeformis和Amphora exigua进行培养实验。In this embodiment, culture experiments were carried out on Amphora coffeaeformis and Amphora exigua.
藻种培养步骤：利用F/2培养基培养藻种；30-38℃静置培养，培养至藻株浓度为0.1g/L。Algae cultivation steps: use F/2 medium to cultivate algae; culture at 30-38°C until the concentration of algae is 0.1g/L.
大量培养步骤：向50升的培养容器中加入40升F/2培养基，F/2培养基盐度为35‰至55‰，将上步骤培养的藻种按藻种培养体积与F/2培养基体积比1：100置于培养容器中，将PC板固定于塑料边框上，形成用于硅藻附着的硅藻附板，多个硅藻附板间隔3厘米垂直置于附板架上，向硅藻附板吹送空气，使海水培养基的温度为35-38℃，对硅藻附板照射灯光，光照强度20μmol.m-2.s-1，使硅藻感受到的昼：夜比为14小时：10小时；Large-scale cultivation steps: Add 40 liters of F/2 culture medium to a 50-liter culture container, the salinity of F/2 culture medium is 35‰ to 55‰, and the algae cultured in the previous step are divided into the culture volume of the algae species and F/2 The medium volume ratio is 1:100 and placed in the culture container, and the PC board is fixed on the plastic frame to form a diatom attachment board for diatom attachment, and multiple diatom attachment boards are vertically placed on the attachment board frame at intervals of 3 cm , blowing air to the diatom attachment plate, so that the temperature of the seawater medium is 35-38 ℃, and irradiating light on the diatom attachment plate, the light intensity is 20μmol.m-2 .s-1 , so that the diatoms feel the day: night The ratio is 14 hours: 10 hours;
采收步骤：在培养容器中培养3至5天后，取出硅藻附板，放置于起膜剂溶液中(添加起膜剂的F/2培养基中，起膜剂添加量为70g/L海水)，1天后，附着于硅藻附板上的硅藻细胞呈膜状或大块片状悬浮在水体中，用网筛过滤采收硅藻细胞；起膜剂由下述重量份数比的组分组成NH3SO4:NaNO3＝5:1。Harvesting steps: After cultivating in the culture container for 3 to 5 days, take out the attached diatom plate and place it in the solution of film-forming agent (in the F/2 medium with film-forming agent added, the amount of film-forming agent added is 70g/L seawater ), after 1 day, the diatom cells attached to the diatom attached plate are suspended in the water body in the form of a film or a large sheet, and the diatom cells are collected by filtering with a mesh sieve; Component composition NH3 SO4 :NaNO3 =5:1.
低温刺激步骤：将采收后的硅藻细胞集中起来，置于18-20℃循环水浴锅中给予低温刺激1天，然后将硅藻细胞冷冻干燥。Low-temperature stimulation step: collect the harvested diatom cells, place them in a circulating water bath at 18-20°C and give low-temperature stimulation for 1 day, and then freeze-dry the diatom cells.
对于双眉藻，海水培养基的温度为35-38℃，本发明的硅藻培养方法的其他实施例中，也可以不加温。For the diatom algae, the temperature of the seawater culture medium is 35-38° C., and in other embodiments of the diatom cultivation method of the present invention, the temperature may not be heated.
这种起膜剂采收硅藻的方法适用于以塑料板或玻璃板作为附板时，而用纱网作为附板时所培养的硅藻附着较紧，密度高，不必刮取采收，可将网(附板)直接投于海参等动物养殖池中，待动物舔食干净后，可将网(附板)再重复利用。The method of harvesting diatoms by this film-forming agent is suitable for when plastic or glass plates are used as attached plates, and when gauze is used as attached plates, the cultivated diatoms are tightly attached and have high density, so there is no need to scrape and harvest them. The net (attached board) can be directly thrown into the animal breeding ponds such as sea cucumbers, and the net (attached board) can be reused after the animals have licked it clean.
本发明的硅藻培养方法包括低温刺激步骤，经分析，低温刺激后的藻细胞油脂含量可提高50％左右，EPA含量可提高3倍左右。The diatom cultivation method of the present invention includes a low-temperature stimulation step. After analysis, the oil content of the algal cells after the low-temperature stimulation can be increased by about 50%, and the EPA content can be increased by about 3 times.
由于硅藻个体微小，从藻液中采收藻细胞一直是个瓶颈。有资料表明，硅藻采收的成本占其养殖成本(包括培养和采收)的20％至30％。寻求一种高效率、低成本、无污染的采收方法是当前硅藻产业亟需解决的问题。Harvesting algal cells from algal liquid has always been a bottleneck due to the tiny size of diatoms. Some data show that the cost of diatom harvesting accounts for 20% to 30% of its cultivation costs (including cultivation and harvesting). Seeking a high-efficiency, low-cost, and pollution-free harvesting method is an urgent problem to be solved in the current diatom industry.
本发明的技术方案，利用底栖硅藻的附着的生物学特性，将细胞培养在固体培养材料上，经起膜剂作用后，附着于固体材料上的硅藻细胞会脱离附着基质，通过网筛过滤即可采收，免除了刮取附着细胞的过程，也节约了一般浮游硅藻离心脱水时所需能耗。The technical scheme of the present invention utilizes the attached biological characteristics of benthic diatoms to culture the cells on the solid culture material. After acting as a film agent, the diatom cells attached to the solid material will break away from the attachment matrix and pass through the net. It can be harvested by sieving and filtering, which avoids the process of scraping off the attached cells, and also saves the energy required for the centrifugal dehydration of general planktonic diatoms.
本发明针对传统的硅藻液体养殖中条件苛刻、能耗高、易污染和细胞密度低的问题，提供一种硅藻培养方法，使用本发明的方法培养硅藻不需要严苛的条件，能耗较低，不易发生污染，并且能够产生高密度的藻细胞。Aiming at the problems of harsh conditions, high energy consumption, easy pollution and low cell density in traditional diatom liquid cultivation, the present invention provides a diatom cultivation method. Using the method of the present invention to cultivate diatoms does not require harsh conditions and can Low consumption, less prone to pollution, and capable of producing high densities of algal cells.
以上仅是本发明的优选实施方式，应当指出，对于本技术领域的普通技术人员来说，在不脱离本发明原理的前提下，还可以作出若干改进和润饰，这些改进和润饰也应视为本发明的保护范围。The above are only preferred embodiments of the present invention, and it should be pointed out that for those of ordinary skill in the art, without departing from the principle of the present invention, some improvements and modifications can also be made, and these improvements and modifications should also be regarded as protection scope of the present invention.
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