A kind of micro-algae compound living body bait for shellfish hatchery and production method thereof
Technical field
The invention belongs to technical field of microalga biology, particularly a kind of micro-algae compound living body bait for shellfish hatchery and production method thereof.
Background technology
The share that the aquaculture of the economic shellfishes such as oyster, mussel, clam and scallop is shared in world's fish production is increasing, has already become one of pillar in living marine resources industry.Micro-algae, as the best bait of shellfish seedling, is the unique suitable food source of shellfish hatchery.
In recent years, along with the fast development of the process of industrialization of various countries' culture fishery, rapidly, the applicating and exploitation of micro-algae in aquaculture feed is developed rapidly the development of Mirco algal food production technology, and the ratio of Mirco algal food shared by aquatic feeds is also increasing.In shellfish seed and breeding production process, the effect of micro-algae mainly contains three types: (1) through specially treated concentrate or the microalgae cell of High Density Cultivation directly as the fresh aquatic food bait of the seedling such as shellfish; (2) using the form of the former powder of micro-algae as high-grade feed additive, special Mirco algal food is researched and developed according to the nutritional need of different breeding kind, the composition of different microalgae nutritional labeling; (3) utilize the Principles of Ecosystem of " micro-algae one zooplankter " food chain, utilize the protozoans such as the microdisk electrode wheel animalcule of Different Nutrition type, halogen worm, and then the high-grade bait indirectly of exploitation.Evaluate the method for Mirco algal food effect, mainly micro-algae is directly carried out as bait test of throwing something and feeding, compare survival rate and the incidence of disease of shellfish; To analyze and research the content of biochemical component of micro-algae and proportion of composing, contrast with the nutritional need of shellfish; Or the bait effect of various micro-algae that two kinds of methods combining are got up more comprehensively and objectively to analyze and research.
At present, the Aguaro Mirco algal food of U.S.'s exploitation utilizes the proportions of different microalgae, can meet the demand of different breeding kind; The Cell System company of Britain develops the micro-algae feed being rich in the high unsaturated fatty acids such as EPA or DHA; Japan develops multiple concentrated Mirco algal food, can need to produce in batches according to the difference of fish, shrimp, crab, shellfish etc.But growth characteristics cause the industrialization degree of its large-scale production still very low specific to micro-algae, up to the present, only have design and the Application comparison success thereof of open culture systems, be amplified to 180,000 m of TaiWan, China
2(producing chlorella) and Mexico City produce 200,000 m of spirulina
2and even, large 50 hectares of arriving western australia biotech company.But open system ubiquity growth parameter(s) and is difficult to control, and culture environment is unstable; Easily contaminated, be difficult to realize sterile culture, the shortcomings such as productive rate is lower, can only be used for spirulina, and the minorities such as chlorella and salt algae can tolerate the microdisk electrode of extreme environment growth; Be difficult to be suitable for the large-scale culture of the economic algae such as Chaetoceros, chrysophyceae, the exploitation of Mirco algal food goods and commercialization are very restricted.
Summary of the invention
Technical problem to be solved by this invention is to provide a kind of technique micro-algae compound living body bait for shellfish hatchery simple, time saving and energy saving, with low cost and production method thereof.
For solving the problems of the technologies described above, the technical solution used in the present invention is as follows:
For micro-algae compound living body bait of shellfish hatchery, the micro-algae algae liquid mixed in number of cells ratio after cultivating by following micro-algae algae kind forms: Chaetoceros muelleri (15%-20%), cocolith (20%-25%), Isochrysis galbana (5%-10%), haematococcus pluvialis (15%-20%), chlorella pyrenoidosa (25%-30%) and P latymonas subcordifomis (10%-15%), and gross density is 1 × 10
5-5 × 10
5individual cell/mL.
For a production method for micro-algae compound living body bait of shellfish hatchery, comprise the steps:
(1) algae kind is selected:
According to the nutritional need of shellfish seedling, micro-algae algae kind is selected to be Chaetoceros muelleri, cocolith, Isochrysis galbana, haematococcus pluvialis, chlorella pyrenoidosa and P latymonas subcordifomis; Described shellfish seedling is Pacific oyster seedling (the eyebot larvae phase);
(2) microdisk electrode:
Utilize bioreactor (ZL201220434882.9) to cultivate respectively the algae kind selected in step (1), obtain micro-algae algae liquid that density is 1 × 109-5 × 1010 cell/mL;
(3) preparation of micro-algae compound living body bait:
By the micro-algae algae liquid obtained in step (2), in number of cells ratio carry out being mixed with for shellfish seedling, gross density is micro-algae compound living body bait of 1 × 105-5 × 105 cell/mL.Micro-algae compound living body bait formula is as follows:
(4) thrown something and fed by the micro-algae compound living body bait obtained in step (3) shellfish seedling, density of throwing something and feeding is 0.1-0.3L/m
3breeding water body.
The invention has the beneficial effects as follows:
(1) microalgae cell is made the nutritive loss that living body bait not only can be avoided because drying causes, and the survival rate of shellfish seedling can be improved, reduce its incidence of disease.
(2) the micro-algae compound living body bait be made up of multiple microalgae cell can the nutrition that causes because of single micro-algae of quasi-complement unbalanced, improve growth rate and the quality of shellfish seedling.
(3) micro-algae compound living body bait that the present invention develops has the features such as cost is low, purity is high, effective, and is easy to realize industrial amplification production.
Detailed description of the invention
According to following embodiment, the present invention may be better understood.But those skilled in the art will readily understand, the content described by embodiment only for illustration of the present invention, and should can not limit the present invention described in detail in claims yet.
Embodiment:
(1) according to the nutritional need of Pacific oyster seedling (eyebot larvae phase), Chaetoceros muelleri, cocolith, Isochrysis galbana, haematococcus pluvialis, chlorella pyrenoidosa and P latymonas subcordifomis is selected to be micro-algae algae kind.
(2) utilize bioreactor (ZL201220434882.9) to cultivate the micro-algae algae kind selected in step (1), obtain micro-algae algae liquid.Wherein the cultivation temperature of Chaetoceros muelleri, cocolith, Isochrysis galbana, haematococcus pluvialis and P latymonas subcordifomis is 23 DEG C, and inoculum density is 1 × 10
4individual cell/mL, culture medium is respectively f/2, NMB3, f/2, BBM and Kang Wei side's culture medium; Chlorella pyrenoidosa cultivation temperature is 28 DEG C, and inoculum density is 1 × 10
4individual cell/mL, culture medium is BG11 culture medium.After bioreactor is cultivated, obtaining density is respectively 1.0 × 10
9(Chaetoceros muelleri), 2.7 × 10
10(cocolith), 1.5 × 10
10(Isochrysis galbana), 6.8 × 10
9(haematococcus pluvialis), 5.0 × 10
10(chlorella pyrenoidosa), 9.2 × 10
9micro-algae algae liquid of (P latymonas subcordifomis) individual cell/mL.
(3) the micro-algae algae liquid will obtained in step (2), by microalgae cell number proportional arrangement become to be used for Pacific oyster seedling, micro-algae compound living body bait of different gross density.
In embodiment 1-12, microalgae cell number ratio design parameter is as shown in table 1.For evaluating the beneficial effect of micro-algae compound living body bait, with the survival rate of Pacific oyster seedling (%) and the incidence of disease (%) for index, the effect evaluating micro-algae compound living body bait is as shown in table 2.
Table 1
Table 2
Comparative example 1:
(1) Chaetoceros muelleri is selected to be micro-algae algae kind.
(2) utilize bioreactor (ZL201220434882.9) to cultivate the micro-algae algae kind selected in step (1), obtain micro-algae algae liquid.The cultivation temperature of Chaetoceros muelleri is 23 DEG C, and inoculum density is 1 × 10
4individual cell/mL, culture medium is f/2 culture medium.Cultivate through bioreactor, obtaining culture density is 1.5 × 10
9micro-algae algae liquid of individual cell/mL.
(3) the micro-algae algae liquid will obtained in step (2), be mixed with for Pacific oyster seedling, gross density is 1.5 × 10
5the microalgae living body bait of individual cell/mL.
(4) the microalgae living body bait obtained in step (3) is pressed 0.3L/m
3water body is thrown something and fed Pacific oyster seedling.
(5) through evaluating, the Pacific oyster seedling 30d utilizing this product to feed, survival rate is 85.5%, and the incidence of disease is 23.6%.
Comparative example 2:
(1) chlorella pyrenoidosa is selected to be micro-algae algae kind.
(2) utilize bioreactor (ZL201220434882.9) to cultivate the micro-algae algae kind selected in step (1), obtain micro-algae algae liquid.The cultivation temperature of chlorella pyrenoidosa is 28 DEG C, and inoculum density is 1 × 10
4individual cell/mL, culture medium is BG11 culture medium.Cultivate through bioreactor, obtaining culture density is 4.2 × 10
10micro-algae algae liquid of individual cell/mL.
(3) the micro-algae algae liquid will obtained in step (2), be mixed with for Pacific oyster seedling, gross density is 4.2 × 10
5the microalgae living body bait of individual cell/mL.
(4) the microalgae living body bait obtained in step (3) is pressed 0.2L/m
3water body is thrown something and fed Pacific oyster seedling.
(5) through evaluating, the Pacific oyster seedling 30d utilizing this product to feed, survival rate is 80.3%, and the incidence of disease is 26.1%.
Visible, the micro-algae compound living body bait be made up of multiple microalgae cell adopting the application to produce does living body bait than adopting separately a kind of microalgae cell, more can improve the survival rate of shellfish seedling, and reduce its incidence of disease.And the mixed proportion between different microalgae cell has considerable influence to evaluation indexes such as shellfish survival rate and the incidences of disease, this nutriment be mainly rich in due to different microalgae cell has difference, as Diatomeae be rich in aliphatic acid, chlorella is rich in protein etc., and the nutriment required for the Different growth phases of shellfish seedling also can be different, therefore applicable micro-algae compound living body bait need be selected for different shellfish seedling.