CN103693751B - Attapulgite powder and rice husk make the holothruian cultures Water quality restoration agent of carrier and method for making thereof and purposes - Google Patents

Attapulgite powder and rice husk make the holothruian cultures Water quality restoration agent of carrier and method for making thereof and purposes Download PDF

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CN103693751B
CN103693751B CN201310713676.0A CN201310713676A CN103693751B CN 103693751 B CN103693751 B CN 103693751B CN 201310713676 A CN201310713676 A CN 201310713676A CN 103693751 B CN103693751 B CN 103693751B
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carrier
rice husk
holothruian cultures
powder
water quality
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CN103693751A (en
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徐永平
尤建嵩
李淑英
李纪彬
周俊龙
徐牧
马欣
高玉平
刘士茹
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DALIAN SEM BIOLOGICAL ENGINEERING TECHNOLOGY CO LTD
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DALIAN SEM BIOLOGICAL ENGINEERING TECHNOLOGY CO LTD
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    • YGENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02ATECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A40/00Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
    • Y02A40/80Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
    • Y02A40/81Aquaculture, e.g. of fish

Abstract

The invention discloses attapulgite powder and rice husk makes the holothruian cultures Water quality restoration agent of carrier and method for making thereof and purposes, the present invention is directed to the physiological habit of Stichopus japonicus and cultivation feature, application Marine Microbial Biotechnology: A Review, by the Bacillus flexus CGMCC of separation screening in holothruian cultures environment? No.5115, bacillus subtilis CGMCC? No.5116 and Bacillus cercus CGMCC? No.5117 is formed, with Semen Maydis powder and bean cake powder for main compost, 28-37 DEG C of fermentation 24-48h, again using attapulgite and rice husk as absorption and suspending carrier, the prepared attapulgite powder of processing and rice husk do the holothruian cultures Water quality restoration agent of carrier. this product is strong to water body environment toleration, and floatability, suspension and sink-float controllability are good, are conducive to the clean-up effect of marine culture water 3 D stereo, it is possible to Hg in adsorbed water body2+、Pb2+、Cr6+、Cd2+、Cu2+Etc. harmful substances such as heavy metal ion, effectively suppress pathogenic microorganism, the generation of prevention Stichopus japonicus disease, improve yield and the quality of holothruian cultures.

Description

Attapulgite powder and rice husk make the holothruian cultures Water quality restoration agent of carrier and method for making thereof and purposes
Technical field
The present invention, in aquaculture and Marine Microbial Biotechnology: A Review field, is specifically related to a kind of attapulgite powder and rice husk makes the holothruian cultures Water quality restoration agent of carrier and method for making thereof and purposes.
Background technology
Sea water is the Essential Environment condition that the precious marine products such as Stichopus japonicus are depended on for existence, as the saying goes: " breeding fish for three points, seven points of foster water ". The quality of water quality directly influences the upgrowth situation of Stichopus japonicus and the economic benefit of raiser. Along with economy and the reach of science, the demand of Stichopus japonicus is increased by people day by day, when natural fish for can not meet the market demand, holothruian cultures be converted into the breeding way of present intensive and batch production already by traditional extensive management, Resource Dependence type breeding way, and apostichopus japonicus culture has become one of pillar industry of Chinese Fishery aquaculture. But; along with Radix Morinae Bulleyanae is intensive and large-scale cultivation develops rapidly; occurring in that some serious problems, constrain the development of its industry, such as remaining bait and Excreta and animal dead volume deposit in water body; decompose and generate a lot of harmful substances; it is degrading water quality, pathogen amount reproduction, causes Stichopus japonicus to produce strong stress; occurring in that shakes the head does not eat, mouth is swollen, health shrink stiff, arrange the phenomenons such as dirty, ulcer in body surface, last body wall large area is rotted and dead. In order to resist Stichopus japonicus disease epidemic disease, long-term and a large amount of abuse of antibiotics and chemical synthetic drug, not only result in the Stichopus japonicus drug resistance to the susceptibility and bacteria pathogeny microorganism etc. of pathogenic microorganism, antibiotic detection also can be made to exceed standard, and cause the pollution of whole ecological environment, had a strong impact on save reduce discharging, food safety and hinder the sustainable development of nuisanceless apostichopus japonicus culture industry.
Attapulgite clay is under the jurisdiction of sepiolite group on mineralogy classification, for the moisture magnesia silicate of layer chain, has the layer chain structure feature of uniqueness, and its desirable chemical molecular formula is: Mg5Si8020(OH)2(OH2)24H2O. Attapulgite owing to having in needle-like, the crystal structure of fibrous agrregate, the features such as its specific surface area is big, chemical stability good, high adsorption capacity, it shows good performance in Adsorption of Heavy Metal Ions, organic pollution, radioelement etc., has a good application prospect in water process.
Rice husk is one layer of shell outside Oryza glutinosa, by inner glume and bigger coetonium form. The long 5-10mm of rice husk, 2.5-5mm, thick 23-30 micron, its color and luster is generally golden yellow, and after crushed, bulk density is up to 384-400kg/m3, as the adjuvant of this product, it is possible to increase the floatability of product, suspension and sink-float controllability, be conducive to the clean-up effect of marine culture water 3 D stereo.
Aquatic product microbial ecological agent, it it is the active bacteria formulation utilizing the beneficial microbe in aquatic animal body or in breeding water body to be formed through special processing process, there is the effect such as ammoxidation, nitrated, denitrification, phosphorus decomposing, sulphur removal and denitrogenation, it is effectively reduced the concentration of the COD in water, ammonia nitrogen, cultured water and sulfide, thus effectively improving water quality. At present, it is applied to the Tiny ecosystem water conditioner of aquaculture, it is only limited to simple microbial inoculum or supported microbial inoculum, there is the inferior positions such as flora Survival Reproduction bad adaptability and quantity minimizing in the former in using, it is sink at the bottom of pond that the latter is substantially major part, centering upper strata water body purifying rate is low, cuts pool bottom environment dissolved oxygen low, is unfavorable for the breeding of microecological microbial agent. So, it is badly in need of that a kind of microbial ecological agent can 3 D stereo be comprehensive can remove holothruian cultures COD in water body, ammonia nitrogen, cultured water and sulfide, Heavy Metals in Waters ion can be removed again, and suppress harmful microbe to breed, reach once to be fully cleaned up the Water quality restoration agent of breeding water body simultaneously.
Summary of the invention
The present invention in order to solve that cultivation water in prior art is seriously polluted, disease takes place frequently, Stichopus japonicus high mortality, Stichopus japonicus heavy metal of body, chemical residual and antibiotic remains the present situation such as exceed standard, and invention attapulgite powder and rice husk make the holothruian cultures Water quality restoration agent of carrier and method for making thereof and purposes. This attapulgite powder and rice husk are done the holothruian cultures Water quality restoration agent of carrier and are mainly made up of bacillus cereus mixing, by bacillus subtilis (Bacillussubtilis) YB-1CGMCCNo.5116, Bacillus cercus (Bacilluscereus) YB-2CGMCCNo.5117 and Bacillus flexus (Bacillusflexus) XH-1CGMCCNo.5115 that separate in holothruian cultures environment and screen, with Semen Maydis powder and bean cake powder for dominant fermentation culture medium, using attapulgite and rice husk as absorption and suspending carrier, it is processed into solid powder. The microbial ecological agent related in the present invention, in 2011 08 month No. 05 deliver to China Committee for Culture Collection of Microorganisms's common micro-organisms center preservation (Institute of Microorganism, Academia Sinica, postcode 100101), address is Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3.
The main aspect of the present invention is: the preparation method that attapulgite powder and rice husk do the holothruian cultures Water quality restoration agent of carrier, including following operating procedure:
1. probiotic bacteria stock solution is prepared
Will on LB agar slant culture-medium well-grown bacillus subtilis YB-1CGMCCNo.5116, Bacillus cercus YB-2CGMCCNo.5117 and Bacillus flexus XH-1CGMCCNo.5115, it is separately added into 2116E culture medium, pH7.0~7.5,33~37 DEG C of concussion cultivation 12~16h, mix with the volume ratio of 1:1:1 again, prepare into probiotic bacteria stock solution;
2. liquid fermentation is cultivated
To adding fermentation liquid in closed fermentation reactor, and add nutrient substance then probiotic bacteria stock solution step 1. prepared by the volume ratio of 0.1~1%, inoculating by the volume ratio of 3-5%, ferment, pressure is 0.1~0.25Mpa, 37 DEG C of fermentation 24h, prepare tunning;
3. spray drying treatment
The tunning of above-mentioned steps 2. gained is sprayed to absorption carrier, dried;
Wherein, sea water LB agar culture medium that step is 1. described and 2116E culture medium, it is culture medium prescription in " Molecular Cloning: A Laboratory guide ";
The 2. described nutrient substance of step is by Semen Maydis powder: bean cake powder: sea water presses (1.5~3): (0.5-1.5): the weight ratio of 100 forms;
Consisting of of the absorption carrier that step is 3. described: attapulgite powder and rice husk mix by the volume ratio of 4:1.
With optimal conditions: in above-mentioned preparation method, the cell concentration of step 1. middle bacteria suspension is each about 1 × 108cfu/ml。
With optimal conditions: in above-mentioned preparation method, in the absorption carrier that step is 3. described, attapulgite powder is 150~200 orders; End-product absorption carrier and probiotic composition, the final concentration of load bacterial content is about 108~1010cfu/g。
With optimal conditions: in above-mentioned preparation method, the weight ratio adding absorption carrier in the product that 3. step prepares is 1:1~1000.
With optimal conditions: in above-mentioned preparation method, step 3. in tunning be spray dried into the method for absorption carrier and be: fermentation culture and vehicle weight ratio is for 1:1~4, and drying time is 12~24h, and product water content is not higher than 5%.
Another aspect of the invention the is in that, profit prepares attapulgite powder with the aforedescribed process and rice husk makes the holothruian cultures Water quality restoration agent product of carrier.
Another aspect of the invention the is in that, utilizes above-mentioned attapulgite powder and rice husk to do the holothruian cultures Water quality restoration agent of carrier application in holothruian cultures.
The holothruian cultures Water quality restoration agent environmental resistance that the attapulgite powder of the present invention and rice husk make carrier is strong, floatability, suspension and sink-float controllability are good, the clean-up effect of the breeding water body 3 D stereo of Stichopus japonicus is notable, especially this attapulgite powder is regularly splashed and rice husk does the holothruian cultures Water quality restoration agent of carrier at the nursery stage of Stichopus japonicus and mature stage, can quickly remove the heavy metal ion in breeding water body and degradation of ammonia nitrogen, nitrite, the harmful substances such as hydrogen sulfide, avoid water acidification and the smelly end, effectively suppress pathogen, alleviate the environment stress to Stichopus japonicus, effectively prevent the generation of Stichopus japonicus disease, improve quality and the yield of holothruian cultures.
Detailed description of the invention
Embodiments of the invention are described in detail below in conjunction with technical scheme.
In embodiment raw materials used by conventional method preparation or by being either commercially available.
LB agar slant culture-medium: containing pancreatin solution peptone 5g, yeast extract 1g, sea crystal 33g, high ferric phosphate 0.1g, agar 15g, 1000mL water, pH7.6 in its composition every liter. Peptone and yeast extract are purchased from Oxoid company of Britain, and agar powder is purchased from Beijing extensive and profound in meaning star biotechnology Co., Ltd, and other raw materials are all purchased from Shanghai Sheng Gong biological engineering limited company.
2216E fluid medium: containing pancreatin solution peptone 5g, yeast extract 1g, high ferric phosphate 0.1g, Chen Haishui 1000mL, pH7.6 in its composition every liter. Peptone and yeast extract are purchased from Oxoid company of Britain, and other raw materials are all purchased from Shanghai Sheng Gong biological engineering limited company.
Semen Maydis powder and bean cake powder, be 100-200 order, be purchased from Wandefu Ind Co., Ltd., Shandong.
Rice husk, is purchased from Dalian Sai Mu group.
Attapulgite powder, is purchased from Jingyuan County, Gansu Province Hao Di industry and trade Co., Ltd, 150-200 order.
The microbial ecological agent related in the embodiment of the present invention includes: included by the bacillus cereus of separation screening in holothruian cultures environment: bacillus subtilis (Bacillussubtilis) YB-1CGMCCNo.5116, Bacillus cercus (Bacilluscereus) YB-2CGMCCNo.5117 and Bacillus flexus (Bacillusflexus) XH-1CGMCCNo.5115.
Three strain bacterial strains be preserved in 2011 08 month No. 05 deliver to China Committee for Culture Collection of Microorganisms's common micro-organisms center (Institute of Microorganism, Academia Sinica, postcode 100101), address is Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3.
Below by example, the present invention is described in further detail.
Embodiment 1:
1. prepare probiotic bacteria stock solution
Will on LB agar slant culture-medium well-grown bacillus subtilis YB-1CGMCCNo.5116, Bacillus cercus YB-2CGMCCNo.5117 and Bacillus flexus XH-1CGMCCNo.5115, it is separately added into 2116E culture medium, pH7.0~7.5,37 DEG C of concussion cultivation 12~16h, mix with the volume ratio of 1:1:1 again, preparing into probiotic bacteria stock solution, cell concentration is about 1 × 108cfu/ml。
2. liquid fermentation is cultivated
To adding fermentation liquid in closed fermentation reactor, and add nutrient substance then probiotic bacteria stock solution step 1. prepared by the volume ratio of 0.1~1%, inoculating by the volume ratio of 3-5%, ferment, pressure is 0.1~0.25Mpa, 37 DEG C of fermentation 24h, prepare tunning;
Above-mentioned nutrient substance is by Semen Maydis powder: bean cake powder: sea water presses (1.5~3): (0.5-1.5): the weight ratio of 100 forms.
3. spray drying treatment
Absorption carrier is 2:1 mixed preparing by attapulgite powder and the rice husk of 150-200 order by volume;
The tunning of above-mentioned steps 2 gained is spray dried in above-mentioned absorption carrier, tunning and vehicle weight are than for 1:4, drying time is 24h, and product water content is not higher than 5%, and prepared attapulgite powder and rice husk do the holothruian cultures Water quality restoration agent of carrier.
Embodiment 2
1. prepare probiotic bacteria stock solution
Will on LB agar slant culture-medium well-grown bacillus subtilis YB-1CGMCCNo.5116, Bacillus cercus YB-2CGMCCNo.5117 and Bacillus flexus XH-1CGMCCNo.5115, it is separately added into 2116E culture medium, pH7.0~7.5,37 DEG C of concussion cultivation 12~16h, mix with the volume ratio of 1:1:1 again, preparing into probiotic bacteria stock solution, cell concentration is about 1 × 108cfu/ml.
2. liquid fermentation is cultivated
To adding fermentation liquid in closed fermentation reactor, and add nutrient substance then probiotic bacteria stock solution step 1. prepared by the volume ratio of 0.1~1%, inoculating by the volume ratio of 3-5%, ferment, pressure is 0.1~0.25Mpa, 37 DEG C of fermentation 24h, prepare tunning;
Above-mentioned nutrient substance is by Semen Maydis powder: bean cake powder: sea water presses (1.5~3): (0.5-1.5): the weight ratio of 100 forms.
3. spray drying treatment
Absorption carrier is 4:1 mixed preparing by attapulgite powder and the rice husk of 150-200 order by volume;
The tunning of above-mentioned steps 2 gained is spray dried in above-mentioned absorption carrier, tunning and vehicle weight are than for 1:4, drying time is 24h, and product water content is not higher than 5%, and prepared attapulgite powder and rice husk do the holothruian cultures Water quality restoration agent of carrier.
Result of use
Choosing the holothruian cultures circle that 3 place's conditions are identical, mean depth 1.5-2m, seedling specification 30-50 head/500g, density is at 9-10 head/m2
No. 1 circle is matched group (not using attapulgite powder and rice husk to do the holothruian cultures Water quality restoration agent of carrier), and 2, No. 3 circles are experimental group (using the holothruian cultures Water quality restoration agent that attapulgite powder and rice husk make carrier); Experimental group is except regularly splashing attapulgite powder described in embodiment 1,2 and rice husk does the holothruian cultures Water quality restoration agent of carrier, and feed mode is the same with matched group with management method.
Experimental period be in JIUYUE, 2013 to October, about 45 days, experimental group was splashed 1 time every 3 days, and the amount of splashing is 5g/m3
During experiment, periodic detection water quality, observe Stichopus japonicus and ingest situation, every 15 all over the world water touch spy, measure Stichopus japonicus body weight. Salinity is 35-38 ‰, and water temperature is 16-23 DEG C, and pH is 7.6-8.5, and dissolved oxygen is not less than 4mg/L.
1. the water quality index comparative result of the holothruian cultures circle of the 45th day after use the present embodiment 1,2
2. use the present embodiment 1,2 impact on Stichopus japonicus growth and immune function in latter 45 days
Head number/m2 Average weight gain amount (g) Respiratory burst vigor Phagocytic rate (%)
Matched group 7.2 4.78 0.132±0.022 36.34±1.73
Embodiment 1 8.1 7.93 0.467±0.012 44.09±0.89
Embodiment 2 8.8 11.64 0.636±0.064 49.55±0.37
Result shows: the attapulgite powder of the present invention and rice husk do the holothruian cultures Water quality restoration agent of carrier, water quality in holothruian cultures circle there is obvious clean-up effect, the content of the harmful substances such as ammonia nitrogen, nitrite, hydrogen sulfide can be greatly reduced, stablize water body pH, effectively suppress pathogen, improve the immunity of Stichopus japonicus, the generation of prevention Stichopus japonicus disease, improve quality and the yield of culturing sea cucumbers.

Claims (6)

1. the preparation method that attapulgite powder and rice husk do the holothruian cultures Water quality restoration agent of carrier, including following operating procedure:
1. probiotic bacteria stock solution is prepared
Will on sea water LB agar slant culture-medium well-grown bacillus subtilis YB-1CGMCCNo.5116, Bacillus cercus YB-2CGMCCNo.5117 and Bacillus flexus XH-1CGMCCNo.5115, it is separately added into 2116E culture medium, pH7.0 ~ 7.5,33 ~ 37 DEG C of concussion cultivation 12 ~ 16h, mix with the volume ratio of 1:1:1 again, prepare into probiotic bacteria stock solution;
2. liquid fermentation is cultivated
To adding fermentation liquid in closed fermentation reactor, and add nutrient substance then probiotic bacteria stock solution step 1. prepared by the volume ratio of 0.1 ~ 1%, inoculating by the volume ratio of 3-5%, ferment, pressure is 0.1 ~ 0.25Mpa, 37 DEG C of fermentation 24h, prepare tunning;
3. spray drying treatment
The tunning of above-mentioned steps 2. gained is sprayed to absorption carrier, dried; Wherein, sea water LB agar slant culture-medium that step is 1. described and 2116E culture medium, it is culture medium prescription in " Molecular Cloning: A Laboratory guide ";
The 2. described nutrient substance of step is by Semen Maydis powder: bean cake powder: sea water presses (1.5 ~ 3): (0.5-1.5): the weight ratio of 100 forms;
Consisting of of the absorption carrier that step is 3. described: attapulgite powder and rice husk mix by the volume ratio of 4:1.
2. preparation method according to claim 1, it is characterised in that the absorption carrier that step is 3. described, attapulgite powder is 150 ~ 200 orders.
3. the preparation method according to any one of claim 1 or 2, it is characterised in that: the product that 3. step prepares is absorption carrier and probiotic composition, final concentration of the 10 of load bacterial content8~1010cfu/g。
4. the preparation method according to any one of claim 1 or 2, it is characterized in that, step 3. in tunning be spray dried into the method for absorption carrier and be: fermentation culture and vehicle weight ratio is for 1:1 ~ 4, and drying time is 12 ~ 24h, and product moisture is not higher than 5%.
5. the attapulgite powder that prepared by the method for claim 1 and rice husk do the holothruian cultures Water quality restoration agent of carrier.
6. attapulgite powder as claimed in claim 5 and rice husk do the holothruian cultures Water quality restoration agent of carrier application in holothruian cultures.
CN201310713676.0A 2013-12-20 2013-12-20 Attapulgite powder and rice husk make the holothruian cultures Water quality restoration agent of carrier and method for making thereof and purposes Active CN103693751B (en)

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