CN110880268B - Method for preparing earthworm specimen - Google Patents
Method for preparing earthworm specimen Download PDFInfo
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- CN110880268B CN110880268B CN201911101379.4A CN201911101379A CN110880268B CN 110880268 B CN110880268 B CN 110880268B CN 201911101379 A CN201911101379 A CN 201911101379A CN 110880268 B CN110880268 B CN 110880268B
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- G—PHYSICS
- G09—EDUCATION; CRYPTOGRAPHY; DISPLAY; ADVERTISING; SEALS
- G09B—EDUCATIONAL OR DEMONSTRATION APPLIANCES; APPLIANCES FOR TEACHING, OR COMMUNICATING WITH, THE BLIND, DEAF OR MUTE; MODELS; PLANETARIA; GLOBES; MAPS; DIAGRAMS
- G09B23/00—Models for scientific, medical, or mathematical purposes, e.g. full-sized devices for demonstration purposes
- G09B23/36—Models for scientific, medical, or mathematical purposes, e.g. full-sized devices for demonstration purposes for zoology
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N1/00—Preservation of bodies of humans or animals, or parts thereof
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Abstract
The invention discloses a method for preparing an earthworm specimen. The manufacturing method comprises the following steps: pre-culturing earthworms, soaking the earthworms in an ethanol solution, filling and fixing the soaked earthworms with pentanediol, and performing sealed preservation by using a preservation solution. The method has the advantages of easily obtained materials, no toxic substances such as toluene and the like, environmental protection, safety, simple manufacturing process and suitability for popularization, and the original physiological structure and appearance of the earthworm can be maintained.
Description
Technical Field
The invention belongs to the field of animal specimen preparation methods, and particularly relates to a preparation method of an earthworm specimen.
Background
Earthworms, also called earthworms, are one of the most widely distributed and typical soil macro-animals in soil, and can significantly improve soil microstructure through activities of feeding, excreting, digging cavities and the like, affecting soil material circulation and energy flow, and are called as "ecosystem engineers" and more people need to know and understand. "one specimen prevails over thousand of words" as an important means for the development of biological monitoring, protection, classification, scientific research and teaching, and specimen preparation is of great significance for improving the understanding of the relationship between human beings and organisms, the understanding of the distribution and habit thereof, and the promotion of biological development.
The standard prepared specimen can be preserved for a long time, has wide application, is an important means for authoritatively identifying the types of the earthworms, has no unified method for preparing the earthworm specimens at present, is simply described according to the traditional literature, is operated automatically, is not detailed and systematic enough, and mostly takes formalin, phenol and other reagents as antiseptic materials, and the reagents can cause certain damage to the specimen and a manufacturer, and needs to be perfected and supplemented along with the continuous progress and development of science and technology.
Disclosure of Invention
The invention aims to provide a method for manufacturing earthworm specimens, which has the advantages of simple and convenient operation, low cost, environmental protection and safety, and the manufactured specimens can be stored for a long time and are suitable for large-scale popularization and application.
The technical scheme adopted by the invention is as follows:
the manufacturing method of the earthworm specimen is characterized by comprising the following steps:
(1) earthworm pre-culture: putting the collected earthworms into a container paved with wet grass paper or hay, keeping the temperature and the humidity, placing the earthworms in the dark, stopping feeding, discharging soil in the intestinal tract, and feeding the wet grass paper or agar to fill the intestinal tract;
(2) soaking: putting the pre-cultured earthworms into an ethanol solution with the mass concentration of 8% -12% until mucus is secreted from the back holes of the earthworms;
(3) fixing and filling: injecting pentanediol fixing liquid into the body cavity of the soaked earthworms from the rear two ends of the earthworms, and adjusting the earthworms to be in a natural appearance shape;
(4) and (3) storage: and (3) immersing the earthworms after being fixed and filled into a preserving fluid for sealed preservation, wherein the preserving fluid comprises potassium sorbate, industrial ethanol, glycerol, citric acid, disodium ethylene diamine tetraacetate and water.
Further, in the step (1), the wet paper or agar is fed for 5-7 days at 22-28 ℃ and 60-80% humidity.
Further, in the step (3), the mass concentration of the pentanediol is 3-6%.
Further, in the step (4), the preservation solution comprises 1-3g of potassium sorbate, 100-150mL of industrial ethanol, 4-12mL of glycerol, 10-15g of citric acid, 1g of disodium ethylenediaminetetraacetate and water, and is prepared into a solution with the volume of 1L.
Further, in the step (4), the preservation solution comprises 2g of potassium sorbate, 130mL of industrial ethanol, 8mL of glycerol, 13g of citric acid, 1g of disodium ethylenediaminetetraacetate and water, and is prepared into a solution with the volume of 1L.
The invention has the beneficial effects that:
(1) according to the invention, the earthworms are fixed and filled by the pentanediol, so that the original physiological structure and appearance of the earthworms can be maintained, toxic substances such as toluene and formalin are not contained, the method is environment-friendly and safe, the cost is low, and the prepared specimen can be stored for more than one year.
(2) The specimen preparation process is simple, the content is detailed, the used materials are easy to obtain, and the specimen preparation method is suitable for popularization and application in enterprises and public institutions, universities and scientific research institutions related to agriculture and forestry.
Drawings
FIG. 1 shows the earthworm specimens prepared in the examples.
Detailed Description
A method for preparing an earthworm specimen comprises the following steps:
(5) earthworm pre-culture: putting Lumbricus into container with wet paper or hay at 22-28 deg.C and humidity of 60-80%, stopping feeding in dark, discharging soil in intestinal tract, and feeding wet paper or agar for 5-7 days to fill intestinal tract;
(6) soaking: putting the pre-cultured earthworms into an ethanol solution with the mass concentration of 8% -12% until mucus is secreted from the back holes of the earthworms;
(7) fixing and filling: injecting 3-6% pentanediol fixing solution into the cavity of the soaked earthworm from the rear end side, and adjusting the earthworm to natural appearance;
and (3) storage: and immersing the earthworms after the fixation and filling into a preservation solution for sealed preservation, wherein the preservation solution comprises 1-3g of potassium sorbate, 100-150mL of industrial ethanol, 4-12mL of glycerol, 10-15g of citric acid, 1g of disodium ethylene diamine tetraacetate and water, and the volume of the solution is 1L.
The technical solution of the present invention will be described in further detail by the preferred embodiments of the present invention.
Example 1: method for preparing earthworm specimen
1. Pre-culturing: putting Lumbricus into wet grass paper container laid in advance, keeping temperature at 25 deg.C and humidity at 70%, stopping feeding in dark, discharging soil in Lumbricus intestinal tract, feeding Lumbricus with crushed wet grass paper or agar for 6 days, and filling in Lumbricus intestinal tract.
2. Soaking: and (3) soaking the pre-cultured earthworms in an ethanol solution with the mass concentration of 10% until back holes of the earthworms secrete a large amount of mucus.
3. Filling and fixing: taking out the soaked earthworms, injecting a pentanediol fixing solution with the mass concentration of 4% into the cavity of the earthworms from the side surfaces of the rear two ends by using an injector, and adjusting the earthworms to be in a free moving state.
4. And (3) storage: preparing 1L of preservation solution by using 2g of potassium sorbate, 130mL of industrial ethanol, 8mL of glycerol, 13g of citric acid, 1g of disodium ethylenediamine tetraacetic acid and water, putting the soaked earthworms into the preservation solution, and preserving in a sealed manner, wherein the prepared specimen can be preserved for more than one year.
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.
Claims (4)
1. The manufacturing method of the earthworm specimen is characterized by comprising the following steps:
(1) earthworm pre-culture: putting earthworms into a container paved with wet paper or hay, keeping temperature and humidity, placing in the dark, stopping eating, discharging soil in the intestinal tract, and feeding wet paper or agar to fill the intestinal tract;
(2) soaking: putting the pre-cultured earthworms into an ethanol solution with the mass concentration of 8% -12% until mucus is secreted from the back holes of the earthworms;
(3) fixing and filling: injecting pentanediol fixing liquid into the cavity of the soaked earthworms from the side surface of the rear end, and adjusting the earthworms to be in a natural appearance shape;
(4) and (3) storage: immersing the earthworms after being fixed and filled into a preserving fluid for sealed preservation, wherein the preserving fluid comprises potassium sorbate, industrial ethanol, glycerol, citric acid, ethylene diamine tetraacetic acid disodium salt and water;
in the step (3), the mass concentration of the pentanediol is 3-6%.
2. The method for preparing the earthworm specimen according to claim 1, wherein in the step (1), the earthworm specimen is fed with wet straw paper or agar for 5 to 7 days at a temperature of 22 to 28 ℃ and a humidity of 60 to 80%.
3. The method for preparing the earthworm specimen according to claim 1, wherein in the step (4), the preservation solution comprises 1-3g of potassium sorbate, 100-150mL of industrial ethanol, 4-12mL of glycerol, 10-15g of citric acid, 1g of disodium ethylenediaminetetraacetate and water, and is prepared into a solution with a volume of 1L.
4. The method for preparing an earthworm specimen according to claim 3, wherein in the step (4), the preservation solution comprises a solution prepared from 2g of potassium sorbate, 130mL of industrial ethanol, 8mL of glycerol, 13g of citric acid, 1g of disodium ethylenediaminetetraacetate and water in a volume of 1L.
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| CN201911101379.4A CN110880268B (en) | 2019-11-12 | 2019-11-12 | Method for preparing earthworm specimen |
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| CN201911101379.4A CN110880268B (en) | 2019-11-12 | 2019-11-12 | Method for preparing earthworm specimen |
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| CN110880268B true CN110880268B (en) | 2021-12-28 |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2008112561A1 (en) * | 2007-03-09 | 2008-09-18 | Pfantastic Medical Research Institute, Llc | Combination therapy for alleviating pain-related conditions |
| CN101180958B (en) * | 2007-12-18 | 2010-05-26 | 重庆师范大学 | Processing method before imbedding biological specimen |
| DE102016005077A1 (en) * | 2016-04-27 | 2017-11-02 | Fraunhofer-Gesellschaft zur Förderung der angewandten Forschung e.V. | Sample container for a cryopreserved biological sample, method for producing the sample container, method for monitoring the temperature of a cryopreserved sample |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| JPH033351A (en) * | 1989-05-31 | 1991-01-09 | Sony Corp | Method of forming wiring |
| KR20080014518A (en) * | 2006-08-11 | 2008-02-14 | 에스케이케미칼주식회사 | Electrostatic dissipative polymers and polymer mixtures thereof |
| CN101636649A (en) * | 2007-02-27 | 2010-01-27 | 恰根有限公司 | Fixation of a biological material |
| CN101473812A (en) * | 2009-01-24 | 2009-07-08 | 曹建明 | Antiseptic fixing liquid composition for cadaver sample and preparation method thereof |
| CN106415234A (en) * | 2014-05-28 | 2017-02-15 | 凯杰有限公司 | Fixative composition for cell-comprising liquid samples |
| CN104365581A (en) * | 2014-11-05 | 2015-02-25 | 中山大学 | Novel specimen conservation solution |
| CN104357432A (en) * | 2014-11-27 | 2015-02-18 | 苏州嘉禧萝生物科技有限公司 | Preserving fluid of glutamate decarboxylase |
| CN105360108A (en) * | 2015-12-16 | 2016-03-02 | 贵阳中医学院 | Specimen preservation fluid good in preservation effect and preparation method of specimen preservation fluid |
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Address after: Guangzhou City, Guangdong province 510650 Tianyuan Road No. 808 Patentee after: Institute of ecological environment and soil, Guangdong Academy of Sciences Address before: Guangzhou City, Guangdong province 510650 Tianyuan Road No. 808 Patentee before: GUANGDONG INSTITUTE OF ECO-ENVIRONMENTAL SCIENCE & TECHNOLOGY |
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