CN114711174B - High-density culture method for somatic insects - Google Patents

High-density culture method for somatic insects Download PDF

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Publication number
CN114711174B
CN114711174B CN202210158888.6A CN202210158888A CN114711174B CN 114711174 B CN114711174 B CN 114711174B CN 202210158888 A CN202210158888 A CN 202210158888A CN 114711174 B CN114711174 B CN 114711174B
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chlorella
culture
aeolosoma
extract
growth promoter
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CN114711174A (en
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魏婉蓉
丁国际
唐量
李小伟
衣学文
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University of Shanghai for Science and Technology
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University of Shanghai for Science and Technology
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    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; CARE OF BIRDS, FISHES, INSECTS; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K61/00—Culture of aquatic animals
    • A01K61/40—Culture of aquatic animals of annelids, e.g. lugworms or Eunice
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K10/00—Animal feeding-stuffs
    • A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
    • A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K50/00—Feeding-stuffs specially adapted for particular animals
    • A23K50/80—Feeding-stuffs specially adapted for particular animals for aquatic animals, e.g. fish, crustaceans or molluscs
    • Y—GENERAL 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
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
    • Y02W10/00—Technologies for wastewater treatment
    • Y02W10/10—Biological treatment of water, waste water, or sewage

Abstract

The invention discloses a method for culturing aeolosoma at high density, which comprises the following steps: A. separating and purifying the soma; B. culturing chlorella powder as growth promoter in high density; C. culturing the chlorella extract as a growth promoter in high density; the aeolosoma hemprichi high-density culture method comprises the following steps: 1) The body worms are from activated sludge or biological membranes in the wastewater biological treatment device, and have the capability of quickly adapting to the wastewater environment; 2) The chlorella powder and the chlorella powder extract are used as the accelerator of the artemia, so that the cost is low, the dosage is small, and the environmental impact of the wastewater is avoided; 3) The chlorella powder and the chlorella extract are used as growth promoters, and the chlorella powder is consumed simultaneously in the growth and propagation process of the hemlock beetles, so that secondary pollution is not caused. 4) The aeolosoma are cultured in a large culture container such as a round pot or a water tank, and the environment is more similar to that of an actual sewage treatment device.

Description

High-density culture method for somatic insects
Technical Field
The invention relates to the field of aeolosoma high-density culture, in particular to a aeolosoma high-density culture method.
Background
In the biological wastewater treatment system, large annelids such as fairy worms, somatic worms and the like often exist. Wherein, the artemia not only is helpful to improve the biological treatment performance of the wastewater, but also has high-efficiency residual sludge reduction effect. Since the reduction of excess sludge by micro-animals is an ecologically friendly method, this method has been highly paid attention to in the field of biological wastewater treatment.
One of the key technologies for playing the roles of the artemia in the biological treatment of the wastewater is to establish a low-cost and high-efficiency cultivation method of the aequorin artemia outside the biological treatment system, so that the artemia can be the dominant species in the biological treatment system of the wastewater by inoculating a large amount of cultivated aequorin artemia to the biological treatment system of the wastewater, namely, a manual intervention method. Chinese patent (CN 102499185 a) discloses a method for mass culture of growth and reproduction of red-eye beetles using supernatant produced by ultrasonic crushing of soaked rice. However, the culture method of increasing the growth density and promoting the growth rate of aeolosoma aestivum was still a considerable research topic. In view of the above, the technical problem to be solved by the invention is to provide a culture method capable of promoting the growth of the artemia salina and improving the growth density of the artemia salina.
Disclosure of Invention
The invention aims to provide a method for culturing aeolosoma at high density, which solves the problems in the background technology.
The invention is realized in such a way that the aeoloma high-density culture method comprises the following steps:
A. separating and purifying the soma;
B. culturing chlorella powder as growth promoter in high density;
C. the chlorella extract is used as growth promoter for culturing hemprichi body insects in high density.
The invention further adopts the technical scheme that: the aeolosoma is a microzoon of the phylum of the annelid, which is close to the order Kong Guamao, and comprises aeolosoma erythraea.
The invention further adopts the technical scheme that: in the step A, the method for separating and purifying the aeolosoma comprises the following steps: collecting an activated sludge or biological film sample from a wastewater biological treatment device, sucking a proper amount of the sample into a culture dish, sucking the sample in the culture dish from the sample in the culture dish by using a pipette gun under the magnification of 4-20 times, placing the sample in deionized water, and shaking the culture dish for 10-20 times; placing the culture dish under a physical microscope, sucking the aeoloma from the culture dish solution by using a pipetting gun, transferring the solution into other culture dishes containing deionized water, and shaking the culture dish for 10-20 times; the above procedure was repeated 3 times to obtain the heme's body insects without the presence of other micro-animals.
The invention further adopts the technical scheme that: the chlorella powder in the step B is used as a growth promoter, and the high-density culture method of the aeolosoma hemprichi comprises the following steps: inoculating the purified artemia obtained in step A into a culture vessel containing a culture solution and inoculating a culture medium derived from an active soilSeparating and purifying bacteria from the mud; the culture container is a round pot or a water tank, when the effective volume of the culture container is more than 0.5m 3 Setting a stirring device, wherein the stirring speed is 5-30 rpm; preparing culture solution by 0.1% tryptone, 0.05% yeast extract, 0.05% sodium chloride and 99.8% distilled water; taking chlorella powder as a growth promoter for promoting the reproduction of aeolosoma, wherein the concentration of the chlorella powder is 5-20 mg/L, and the adding frequency is 1-3 times/week; culturing the aequorin at 20-28 deg.c and static or oscillating condition.
The invention further adopts the technical scheme that: the preparation method of the chlorella powder comprises the following steps: drying fresh cultured Chlorella by evaporating, pulverizing dried Chlorella with grinder, sieving with 100 mesh sieve to obtain dried Chlorella powder, and collecting the sieved Chlorella powder as growth promoter for body insects.
The invention further adopts the technical scheme that: the chlorella extract in the step C is used as a growth promoter, and the hemerocallis high-density culture method comprises the following steps: inoculating the purified aeoloma obtained in the step A into a culture container containing a culture solution, and inoculating bacteria separated and purified from the activated sludge; the culture container is a round pot or a water tank, when the effective volume of the culture container is more than 0.5m 3 Setting a stirring device, wherein the stirring speed is 5-30 rpm; preparing culture solution by 0.1% tryptone, 0.05% yeast extract, 0.05% sodium chloride and 99.8% distilled water; taking chlorella extract as a growth promoter for promoting the reproduction of aeolosoma; the volume ratio of the chlorella extract is 0.5% -5%, and the adding frequency is 1-3 times/week; culturing the aequorin at 20-28 deg.c and static or oscillating condition.
The invention further adopts the technical scheme that: the preparation method of the chlorella extract comprises the following steps: preparing chlorella liquid with the concentration of 1-2 g/L from freshly cultured chlorella; the chlorella powder liquid is subjected to water bath for 1 to 2 hours at the temperature of 70 to 90 ℃, and is filtered by a screen or a filter with the size of more than 100 meshes, and the filtered liquid is chlorella extract.
The invention has the beneficial effects that: the aeolosoma hemprichi high-density culture method has the remarkable advantages that: 1) The body worms are from activated sludge or biological membranes in the wastewater biological treatment device, and have the capability of quickly adapting to the wastewater environment; 2) The chlorella powder and the chlorella powder extract are used as the accelerator of the artemia, so that the cost is low, the dosage is small, and the environmental impact of the wastewater is avoided; 3) The chlorella powder and the chlorella extract are used as growth promoters, and the chlorella powder is consumed simultaneously in the growth and propagation process of the hemlock beetles, so that secondary pollution is not caused. 4) The aeolosoma are cultured in a large culture container such as a round pot or a water tank, and the environment is more similar to that of an actual sewage treatment device.
Drawings
FIG. 1 is a graph showing growth of aeolosoma aestivum when chlorella powder is added as a growth promoter in the method for high-density culture of aeolosoma aestivum;
fig. 2 is a graph showing growth of aeolosoma aegerita when chlorella extract is added as a growth promoter in the method for high-density culture of aeolosoma aegerita.
Detailed Description
Embodiment one: FIG. 1 shows a method for achieving high density cultivation of the aeolosoma using Chlorella as a growth promoter, comprising the following steps:
the separation and purification method of the somatic worms comprises the following steps: collecting activated sludge or a biological film sample from a wastewater biological treatment device, sucking a proper amount of the sample into a culture dish, sucking the aeolochia from the sample in the culture dish by a pipetting gun under the magnification of 20 times, placing the sample into deionized water, and shaking the culture dish for 15 times; placing the culture dish under a physical microscope, sucking the aescintillant from the culture dish solution by using a pipetting gun, transferring the aescintillant into other culture dishes containing deionized water, and shaking the culture dish for 15 times; the above procedure was repeated 3 times to obtain the heme's body insects without the presence of other micro-animals.
The preparation method of the chlorella powder liquid comprises the following steps: drying fresh cultured Chlorella by evaporating, pulverizing dried Chlorella with grinder, sieving with 100 mesh sieve to obtain dried Chlorella powder, and collecting the sieved Chlorella powder as growth promoter for body insects.
The chlorella powder is used as a growth promoter for the high-density culture method of aeolosoma hemprichi: inoculating purified aeolosoma into culture container containing culture solution, and inoculating from livingSeparating and purifying bacteria from the sexual sludge; the culture container is a round pot with effective volume of 0.5m 3 The stirring speed of the stirrer is 25rpm; preparing culture solution by 0.1% tryptone, 0.05% yeast extract, 0.05% sodium chloride and 99.8% distilled water; taking chlorella powder as a growth promoter for promoting the reproduction of aeolosoma, wherein the concentration of the chlorella powder is 10mg/L, and the adding frequency is 1-3 times/week; culturing the somatic worms at 25 ℃ under the static condition.
Analysis of somatic insect growth and propagation curves:
as can be seen from FIG. 1, the addition of chlorella powder as a growth promoter (group A) in the round pot, and the absence of the growth promoter (group B) in the round pot, compared with the control group without chlorella powder, the group A is continuously increased in number from the next day and higher than the control group, the population density reaches 180ind/mL until the 5 th day, and the population density of the control group 5 th day is 120ind/mL, which is obviously less than that of the group A; the chlorella powder within a certain concentration range has the obvious effect of promoting the growth and reproduction of the aescida.
Embodiment two: FIG. 2 shows a method for achieving high density cultivation of aeolosoma aestivum by using chlorella extract as growth promoter, comprising the following steps: the separation and purification method of the somatic worms comprises the following steps: collecting activated sludge or a biological film sample from a wastewater biological treatment device, sucking a proper amount of the sample into a culture dish, sucking the aeolochia from the sample in the culture dish by a pipetting gun under the magnification of 20 times, placing the sample into deionized water, and shaking the culture dish for 15 times; placing the culture dish under a physical microscope, sucking the aescintillant from the culture dish solution by using a pipetting gun, transferring the aescintillant into other culture dishes containing deionized water, and shaking the culture dish for 15 times; the above procedure was repeated 3 times to obtain the heme's body insects without the presence of other micro-animals.
The preparation method of the chlorella extract comprises the following steps: preparing chlorella liquid with the concentration of 1g/L from freshly cultured chlorella; the chlorella powder liquid is subjected to water bath at 75 ℃ for 1h, and is filtered by a 100-mesh screen, and the filtered liquid is chlorella extract.
The chlorella extract is used as a growth promoter for hemprichi bulk insect high-density culture method: inoculating purified aeolosoma hemprichi into a culture container containing culture solutionInoculating bacteria separated and purified from the activated sludge; the culture container is a round pot with effective volume of 0.5m 3 The stirring speed of the stirrer is 25rpm; preparing culture solution by 0.1% tryptone, 0.05% yeast extract, 0.05% sodium chloride and 99.8% distilled water; taking chlorella extract as a growth promoter for promoting the reproduction of aeolosoma; the volume ratio of chlorella extract is 2%, and the adding frequency is 2 times/week; culturing aeolosoma under the condition of standing at 25 ℃;
analysis of somatic insect growth and propagation curves:
as can be seen from FIG. 2, the addition of chlorella powder as a growth promoter in the round pot (group A) and the absence of the growth promoter in the round pot (group B) resulted in a continuous increase in the number of the grown insects from the next day and higher than the control group, the population density reached 202ind/mL on day 5, and the population density of the grown insects was 126ind/mL on day 5, which was significantly lower than that of the control group A. The chlorella extract with a certain concentration range has the obvious effect of promoting the growth and reproduction of the aeolosoma aegerita.
The foregoing description of the preferred embodiments of the invention is not intended to be limiting, but rather is intended to cover all modifications, equivalents, and alternatives falling within the spirit and principles of the invention.

Claims (1)

1. The method for culturing the aeolosoma in high density is characterized by comprising the following steps of:
A. separating and purifying the soma;
B. culturing chlorella powder as growth promoter in high density;
C. culturing the chlorella extract as a growth promoter in high density;
the aeolosoma is a micromanipulation of the phylum of the annelid, which is approximately Kong Guamao meshes of the somatidae, and comprises aeolosoma erythraea;
in the step A, the method for separating and purifying the aeolosoma comprises the following steps: collecting an activated sludge or biological film sample from a wastewater biological treatment device, sucking a proper amount of the sample into a culture dish, sucking the artemia from the sample in the culture dish by a pipetting gun under the magnification of 4-20 times, placing the artemia into deionized water, and shaking the culture dish for 10-20 times; placing the culture dish under a physical microscope, sucking the somatic insects out of the culture dish solution by using a pipetting gun, transferring the somatic insects into other culture dishes containing deionized water, and shaking the culture dish for 10-20 times; repeating the separation and purification process of the soma for 3 times to obtain the soma hemsleyanum without other micro animals;
the chlorella powder in the step B is used as a growth promoter, and the high-density culture method of the aeolosoma hemprichi comprises the following steps: inoculating the purified artemia obtained in step a to a culture vessel containing a culture solution and inoculating bacteria isolated and purified from the activated sludge; the culture container is a round tank or a water tank, and a stirring device is arranged when the effective volume of the culture container is larger than 0.5m, and the stirring speed is 5-30 rpm; preparing culture solution by 0.1% tryptone, 0.05% yeast extract, 0.05% sodium chloride and 99.8% distilled water; taking chlorella powder as a growth promoter for promoting the propagation of aeolosoma aestivum, wherein the concentration of the chlorella powder is 5-20 mg/L, and the adding frequency is 1-3 times/week; culturing the culture pond insects at the temperature of 20-28 ℃ under the condition of standing or shaking;
the preparation method of the chlorella powder comprises the following steps: drying fresh cultured Chlorella by evaporation, pulverizing dried Chlorella with grinder, sieving with 100 mesh sieve to obtain dried Chlorella powder, and collecting the sieved Chlorella powder as growth promoter for body insects;
the chlorella extract in the step C is used as a growth promoter, and the hemerocallis high-density culture method comprises the following steps: inoculating the purified aeoloma obtained in the step A into a culture container containing a culture solution, and inoculating bacteria separated and purified from the activated sludge; the culture container is a round tank or a water tank, and a stirring device is arranged when the effective volume of the culture container is larger than 0.5m, and the stirring speed is 5-30 rpm; preparing culture solution by 0.1% tryptone, 0.05% yeast extract, 0.05% sodium chloride and 99.8% distilled water; taking chlorella extract as a growth promoter for promoting the reproduction of aeolosoma; the volume ratio of the chlorella extract is 0.5% -5%, and the adding frequency is 1-3 times/week; culturing the culture pond insects at the temperature of 20-28 ℃ under the condition of standing or shaking;
the preparation method of the chlorella extract comprises the following steps: preparing chlorella liquid with the concentration of 1-2 g/L from freshly cultured chlorella; and (3) carrying out water bath on the chlorella liquid for 1-2 hours at 70-90 ℃, and filtering the chlorella liquid by using a screen or a filter with a mesh of more than 100 meshes to obtain a filtered liquid, namely the chlorella extract.
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Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102180574A (en) * 2011-06-14 2011-09-14 上海大学 Surplus sludge reduction method combining ultrasonic treatment with microfauna ingestion
CN102499185A (en) * 2011-10-26 2012-06-20 同济大学 Method for cultivating aeolosoma hemprichi on large scale

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR101005388B1 (en) * 2005-06-14 2010-12-30 아사히 가세이 케미칼즈 가부시키가이샤 Apparatus for Water Treatment and Method of Treating Water

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102180574A (en) * 2011-06-14 2011-09-14 上海大学 Surplus sludge reduction method combining ultrasonic treatment with microfauna ingestion
CN102499185A (en) * 2011-10-26 2012-06-20 同济大学 Method for cultivating aeolosoma hemprichi on large scale

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
具有促进污水生物处理系统中有益微型动物生长功能的生物活性物质探索;杨芳芳;《万方学位论文电子期刊》;20101222;48-49 *

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