CN109300376A - A kind of method of skeleton specimen fleshing - Google Patents
A kind of method of skeleton specimen fleshing Download PDFInfo
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- CN109300376A CN109300376A CN201811269476.XA CN201811269476A CN109300376A CN 109300376 A CN109300376 A CN 109300376A CN 201811269476 A CN201811269476 A CN 201811269476A CN 109300376 A CN109300376 A CN 109300376A
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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
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Abstract
The invention discloses a kind of methods of skeleton specimen fleshing.Method includes the following steps: rejecting the soft tissue of animal carcass 85%~90%, retains bone portion, bone is then divided into four limbs, skull, thorax, lumbar vertebrae and hipbone part, is encapsulated respectively；Protease, degreasing agent and water are subjected to water bath with thermostatic control with after mixed solution by 20~50:1~2:2000~2100 mass ratio；Packaged bone is respectively put into mixed solution, so that mixed solution just floods bone, impregnates 4~5 h, soft tissue is washed away with high pressure water, rejects the impurity on bone；Bone is put into mixed solution again, is dipped to bone surface free from admixture, bone is taken out, soft tissue is washed away with high pressure water, by bone detergent wash clean.The present invention treated bone is clean and complete, avoids the damage of bone, and fleshing method is simple, and in the case where keeping bone complete situation, there is huge application potential in the shortening fleshing time.
Description
Technical field
The invention belongs to sample preparation technical fields.In particular it relates to a kind of method of skeleton specimen fleshing.
Background technique
Zoological specimens occupy especially important status in biology scientific research, teaching and work of popular science, and skeleton specimen is
Studying the morphosis of animal skeleton and its main material with other organ correlations, animal skeleton specimen not only has exhibition
The function of showing, and also there is extremely important role in traditional morphological taxonomy and archaeology.With the hair of science of heredity
Exhibition, researcher generally require to classify in terms of form and heredity two, this just needs the skeleton specimen of various animals to be used for
Data support that therefore, the production of animal skeleton specimen is particularly important.
The sample and skeleton specimen manufacturing technology of animal skeleton also see some textbooks and magazine article, traditional bone
Slide processing generally comprises: fleshing, go marrow, burn into bleaching and etc., manufacturing process is cumbersome, especially fleshing step,
It heavy workload and needs operator that there is stronger professional knowledge and operative skill, otherwise will cause the damage of bone, production week
Phase is long.
In existing bone fleshing method, most of is to bite method etc. using boiling method, strong acid and strong base method, worm and gone
Meat, these methods highlight many deficiencies in practical application, such as boiling method overlong time and are easily damaged bone, strong acid
Highly basic has certain risk, and the worm method of biting is more suitable for meiofauna bone.Furthermore each position skeletal structure of animal body is different,
The constituent of soft tissue is not limited only to muscle etc., and these method universalities are poor at present, treated in aforementioned manners bone
Bone often damages, and fleshing is not clean.In addition, traditional bone production method is primarily adapted for use in, figure is relatively large to be moved
Object, practicability is lower, and application range compares limitation, so cannot be widely used by people.
Summary of the invention
The technical problem to be solved by the present invention is to overcome above-mentioned the deficiencies in the prior art, a kind of skeleton specimen fleshing is provided
Method, by using the alkali protease and degreasing agent of rational proportion, constant temperature impregnates the bone for removing most of soft tissue, uses this
Method treated bone is clean and complete, avoids the damage of bone.
Above-mentioned purpose of the present invention is achieved through the following technical solutions:
A kind of method of skeleton specimen fleshing, comprising the following steps:
S1. the soft tissue for rejecting animal carcass 85%~90%, retains bone portion, bone is then divided into four limbs, skull, chest
Wide, lumbar vertebrae and hipbone part, encapsulate respectively；
S2. it after protease, degreasing agent and water being pressed 20~50:1~2:2000~2100 quality than mixed solution, carries out permanent
Tepidarium；
S3. the packaged bone of step S1 is respectively put into the mixed solution of step S2, so that mixed solution just floods bone
Bone impregnates 4~5 h, washes away soft tissue with high pressure water, then reject the impurity on bone；
S4. the bone of step S3 is put into the mixed solution of step S2, is dipped to bone surface free from admixture, take out bone, used
High pressure water washes away soft tissue, by bone detergent wash clean.
The soft tissue refers to internal organ, muscle, grease.
The water is clear water.
The high pressure water, which refers to, is forced into 10~25 MPa for water by high pressure water generating device, then by with Small Holes
The injection apparatus of diameter is converted to fine " water jet " of high speed.
Preferably, in step S2, the mass ratio of protease, degreasing agent and water is 30~45:1~2:2000~2050.
It is highly preferred that the mass ratio of protease, degreasing agent and water is 40:1:2000 in step S2.
Preferably, in step S2, the concentration of the protease is 0.95%~2.5%.
It is highly preferred that the concentration of the protease is 1.5%~2.5% in step S2.
Most preferably, in step S2, the concentration of the protease is 2%.
Preferably, in step S2, the concentration of the degreasing agent is 0.05%~0.1%.
It is highly preferred that the concentration of the degreasing agent is 0.05% in step S2.
Preferably, in step S2, the protease in alkali protease, acid protease, neutral proteinase one
Kind is several；The degreasing agent is selected from one or more of alkaline defatting agent, lotion degreasing agent, solvent degreasing agent.
It is highly preferred that the protease is alkali protease in step S2；The degreasing agent is alkaline defatting agent.
It is further preferred that the alkali protease is that Novi believes protease；The alkaline defatting agent is fur degreasing
Agent.
Preferably, in step S2, the condition of the water bath with thermostatic control are as follows: in 55~60 DEG C of 4~5 h of water bath with thermostatic control.
Preferably, in step S3, the time of the immersion is 4 h.
Preferably, in step S3, the impurity on the bone includes ligament, spinal cord and brain.
Preferably, in step S4, the time of immersion is 1~3 h.
It is highly preferred that the time of immersion is 2 h in step S4.
Preferably, in step S4, the concentration of the detergent is 1.0%~2%.
It is highly preferred that the concentration of the detergent is 1.0% in step S4.
Preferably, in step S4, the detergent be powder detergent, liquid detergent or solid detergent one kind or
It is several.
It is highly preferred that the detergent is liquid detergent in step S4.
Preferably, the four limbs, skull, thorax, lumbar vertebrae, hipbone total soaking time be respectively 4~8 h, 4~5 h, 4
~6 h, 4~6 h, 4~6 h.
It is highly preferred that total soaking time of the four limbs, skull, thorax, lumbar vertebrae, hipbone is respectively 7 h, 5 h, 6 h, 6
h、6 h。
The bone of different parts takes out clear water by different period and rinses, and with the method, treated that bone is clean and complete
It is whole, avoid the damage of bone.
Bone fleshing method of the invention is suitable for the various larger animals such as pig, dog, cat and meiofauna skeleton specimen
Production, it is easy to operate, it is easy to use；The fleshing method shortens the time of sample fleshing, while fleshing works well, and can guarantee
Each portion's bone is completely complete, can completely save the shape of bone, saves the time of staff, simple, convenient.
Compared with prior art, the beneficial effects of the present invention are:
The present invention utilizes the alkali protease and degreasing agent of rational proportion, and constant temperature impregnates the bone for removing most of soft tissue, uses
The method treated bone is clean and complete, avoids the damage of bone, and fleshing method is simple, is not necessarily to a large amount of manpower and material resources,
Shorten the fleshing time in the case where keeping the complete situation of sample bone, convenient for observation, provides convenience for research and teaching etc., and should
Bone fleshing method is easy to operate, and the period is shorter, and common scientific and technical personnel can make.
Specific embodiment
Further illustrate the present invention below in conjunction with specific embodiment, but embodiment the present invention is not done it is any type of
It limits.Unless stated otherwise, the present invention uses reagent, method and apparatus is the art conventional reagents, method and apparatus.
Unless stated otherwise, following embodiment agents useful for same and material are commercially available.
Novi believes that protease is that Novi believes protease 1.6(Protamex 1.6)；Fur fat removal agent is bought in Shandong Guan County
Li Run daily use chemicals Co., Ltd.
A kind of method of the skeleton specimen fleshing of embodiment 1
A kind of method of skeleton specimen fleshing, comprising the following steps:
(1) soft tissue that animal carcass 85%~90% is rejected using scalpel, is retained bone portion, is then divided into four through bone
Limb, skull, thorax, lumbar vertebrae and hipbone part, are encapsulated with mesh bag respectively；
(2) configure solution: believe protease: Fur fat removal agent by Novi: clear water=40:1:2000 mass ratio prepares solution；It should
Solution is put in 4 h in 60 DEG C of thermostat water baths, until solution temperature is constant；Wherein, the concentration of Novi's letter protease is 2%, fur
The concentration of degreasing agent is 0.05%；
(3) the packaged bone of step (1) is put into above-mentioned solution, so that solution just floods bone, it should be noted that
Be skull since bone is than relatively thin, do not need the too long of time, therefore skull is put in container topmost；It is taken out after impregnating 4 h
Portions of soft tissue is washed away with pressure hydraulic giant, is largely rejected the impurity such as ligament, spinal cord, brain above bone with scalpel,
Bone major part bone need to be separated, then be strung vertebra with filament by especially extremity portion, joint part.
(4) bone is put into solution again, soft tissue removal effect is observed after 2 h；Continue to disappear if ineffective
Change, until bone surface takes out bone, washes away soft tissue with pressure hydraulic giant without obvious impurity, it is 1.0% by bone concentration
Complete, clean bone can be obtained in detergent wash clean.
A kind of method of the skeleton specimen fleshing of embodiment 2
A kind of method of skeleton specimen fleshing, comprising the following steps:
(1) soft tissue that animal carcass 85%~90% is rejected using scalpel, is retained bone portion, is then divided into four through bone
Limb, skull, thorax, lumbar vertebrae and hipbone part, are encapsulated with mesh bag respectively；
(2) configure solution: press alkali protease: alkaline defatting agent: clear water=30:1:2050 mass ratio prepares solution；This is molten
Liquid is put in 5 h in 55 DEG C of thermostat water baths, until solution temperature is constant；Wherein, the concentration of alkali protease is 1.5%, and alkalinity is de-
The concentration of rouge agent is 0.1%；Alkali protease and alkaline defatting agent is commercially available obtains；
(3) the packaged bone of step (1) is put into above-mentioned solution, so that solution just floods bone, it should be noted that
Be skull since bone is than relatively thin, do not need the too long of time, therefore skull is put in container topmost；It is taken out after impregnating 5 h
Portions of soft tissue is washed away with pressure hydraulic giant, is largely rejected the impurity such as ligament, spinal cord, brain above bone with scalpel,
Bone major part bone need to be separated, then be strung vertebra with filament by especially extremity portion, joint part.
(4) bone is put into solution again, soft tissue removal effect is observed after 1 h；Continue to disappear if ineffective
Change, is 2% to wash by bone concentration until bone surface without obvious impurity, takes out bone, washes away soft tissue with pressure hydraulic giant
Agent wash clean is washed, complete, clean bone can be obtained.
A kind of method of the skeleton specimen fleshing of embodiment 3
A kind of method of skeleton specimen fleshing, comprising the following steps:
(1) soft tissue that animal carcass 85%~90% is rejected using scalpel, is retained bone portion, is then divided into four through bone
Limb, skull, thorax, lumbar vertebrae and hipbone part, are encapsulated with mesh bag respectively；
(2) configure solution: press neutral proteinase: solvent degreasing agent: clear water=45:2:2000 mass ratio prepares solution；This is molten
Liquid is put in 5 h in 60 DEG C of thermostat water baths, until solution temperature is constant；Wherein, the concentration of neutral proteinase is 2.5%, and solvent is de-
The concentration of rouge agent is 0.08%；Neutral proteinase and solvent degreasing agent is commercially available obtains；
(3) the packaged bone of step (1) is put into above-mentioned solution, so that solution just floods bone, it should be noted that
Be skull since bone is than relatively thin, do not need the too long of time, therefore skull is put in container topmost；It is taken out after impregnating 5 h
Portions of soft tissue is washed away with pressure hydraulic giant, is largely rejected the impurity such as ligament, spinal cord, brain above bone with scalpel,
Bone major part bone need to be separated, then be strung vertebra with filament by especially extremity portion, joint part.
(4) bone is put into solution again, soft tissue removal effect is observed after 1 h；Continue to disappear if ineffective
Change, is 2% to wash by bone concentration until bone surface without obvious impurity, takes out bone, washes away soft tissue with pressure hydraulic giant
Agent wash clean is washed, complete, clean bone can be obtained.
A kind of method of the skeleton specimen fleshing of embodiment 4
A kind of method of skeleton specimen fleshing, comprising the following steps:
(1) soft tissue that animal carcass 85%~90% is rejected using scalpel, is retained bone portion, is then divided into four through bone
Limb, skull, thorax, lumbar vertebrae and hipbone part, are encapsulated with mesh bag respectively；
(2) configure solution: press acid protease: lotion degreasing agent: clear water=20:1:2100 mass ratio prepares solution；This is molten
Liquid is put in 5 h in 60 DEG C of thermostat water baths, until solution temperature is constant；Wherein, the concentration of acid protease is 0.95%, lotion
The concentration of degreasing agent is 0.5%；
(3) the packaged bone of step (1) is put into above-mentioned solution, so that solution just floods bone, it should be noted that
Be skull since bone is than relatively thin, do not need the too long of time, therefore skull is put in container topmost；It is taken out after impregnating 5 h
Portions of soft tissue is washed away with pressure hydraulic giant, is largely rejected the impurity such as ligament, spinal cord, brain above bone with scalpel,
Bone major part bone need to be separated, then be strung vertebra with filament by especially extremity portion, joint part.
(4) bone is put into solution again, soft tissue removal effect is observed after 3 h；Continue to disappear if ineffective
Change, is 2% to wash by bone concentration until bone surface without obvious impurity, takes out bone, washes away soft tissue with pressure hydraulic giant
Agent wash clean is washed, complete, clean bone can be obtained.
Influence of the different protease concentrations of embodiment 5 to fleshing effect
1, method: taking dog (8~10 kg), cat (4~6 kg), pig (10~15 kg) corpse each 5, successively using 0.1%,
0.95%, 1.5%, 2%, 2.5% alkali protease carries out fleshing processing as described in Example 1.
2, result
(1) experiment discovery, when alkali protease (Novi believe protease) concentration is in 0.95%~2.5% section, fleshing effect compared with
It is good；Wherein, basic protein enzyme concentration fleshing better effect in 1.5%~2.5% section；If concentration is relatively low, take long time；If
Concentration is higher, although shortening the fleshing time, different degrees of damage occurs in part flat bone edge；Secondly, effect compared with
Good protease concentration section continues to test, and finally found that protease concentration effect at 2% is best.
(2) experiment discovery, 55~60 DEG C at a temperature of, alkali protease cut grease class soft tissue effect it is poor, no
It can sufficiently decompose the soft tissue of bone grease type, and the temperature drift that alkaline lipase needs；Using alkali protease and take off
Both rouge agent synergy quick fleshing and can reduce fat in the case where keeping the complete situation of bone.In addition, the present invention is unexpected
It was found that comprehensive fleshing effect is most when 0.05% degreasing agent (Fur fat removal agent) being added in the protein enzyme solution that concentration is 2%
Good, more clean by the bone of the processing, the fleshing time is short, and bone does not damage.
Influence of the different soaking times of embodiment 6 to fleshing effect
1, method:
(1) 5 tool pigs (10~15 kg) corpse is taken, 5 corpses are removed most of flesh by number 1, No. 2, No. 3, No. 4, No. 5
Bone is divided into four limbs, skull, thorax, lumbar vertebrae, hipbone respectively by meat, is encapsulated with gauze bag；
(2) bone lacing number is put into (control Novi letter egg in the mixed solution of Novi's letter protease, Fur fat removal agent and clear water
White enzyme, Fur fat removal agent, clear water mass ratio be 40:1:2000) impregnate, 55 DEG C of water bath with thermostatic control fleshings, soaking time is respectively 3
H(1 bone), 4 h(2 bones), 5 h(3 bones), 6 h(4 bones), 7 h(5 bones).
Experiment finds that in No. 1 bone, all bones are clean without fleshing；In No. 2 bones, skull upper jaw position is also deposited
There is Portions of soft tissue, but other bones are still clean without fleshing, especially four limbs, the treatment effect of ligament is little；No. 3 bones
In, skull surface is without obvious impurity, but brain tissue is still complete in cranial cavity, the remaining less portion of lumbar vertebra and hipbone part
Divide soft tissue, but still processing is not clean for the ligament of extremity portion；In No. 4 bones, skull part has already appeared occipital bone and falls off
Sign, Brain tissue volume become smaller, however it remains in cranial cavity, dental part falls off, and thorax, lumbar spinal cord still have, hipbone
Part is without obvious impurity；In No. 5 bones, skull part occipital bone is completely fallen off, and brain tissue falls off, and tooth etc. is most of de-
It falls, the disc material of mandibular dimidiation, lumbar vertebrae, thorax falls off, hipbone dimidiation, and the small coccyx in part occurs slight
Damage, but the articular portion ligament of four limbs is again without completely falling off.
Above step is disposably to impregnate, and tests discovery time when 4 h or so, skull surface basic handling
Completely, but brain still has, 6 h or so, and thorax, lumbar vertebrae, hipbone spinal cord still have, and extremity portion does not have in 7 h
Processing is clean, therefore, improves soaking time again, prepares the little pig corpse of a tool size gap again, and fleshing impregnates 4 h,
Skull surface has been handled completely at this time, all take out, giant rinse bone, with scalpel by above four limbs ligament,
The impurity such as spinal cord, brain are largely rejected, and especially bone major part bone need to be separated, then be used by extremity portion, joint part
Filament strings vertebra.
Bone is put into solution again, removal effect is observed after 1 h, discovery skull has handled completely, taken out and use
Detergent is rinsed well, and other parts still have part residual, is observed after 1 h, finds thorax, lumbar vertebrae, several parts of hipbone
It has handled completely, taking-up is rinsed well with detergent, and extremity portion is due to joint position ligament, and processing still has residual, after 1
Observing effect after h, discovery four limbs have been handled completely, and taking-up is rinsed well with detergent.
In summary: four limbs, skull, thorax, lumbar vertebrae, hipbone total soaking time be respectively 4~8 h, 4~5 h, 4~6
H, 4~6 h, 4~6 h fleshing effects are preferable, and best total soaking time is respectively 7 h, 5 h, 6 h, 6 h, 6 h.
In addition, once having identical effect using animal skeleton specimens such as the fleshing method production cat of the present embodiment, dog, pig, sheep
Fruit, discovery time herein within the scope of, i.e., four limbs, skull, thorax, lumbar vertebrae, hipbone total soaking time be respectively 4~8 h,
4~5 h, 4~6 h, 4~6 h, 4~6 h fleshing effects are preferable, and best total soaking time is respectively 7 h, 5 h, 6 h, 6 h, 6
h。
The different quality of 7 protease of embodiment, degreasing agent and water compares the influence of fleshing effect
1, method:
(1) 9 tool dog corpse are taken, the method for reference implementation example 1 handles bone after numbering respectively, wherein other conditions and embodiment 1
Method it is identical, observation Novi letter protease, Fur fat removal agent and water different quality compare the influence of fleshing effect.
The different quality of 1 protease of table, degreasing agent and water compares the influence of fleshing effect
Experiment discovery, protease and degreasing agent joint are used for fleshing, and the mass ratio of protease, degreasing agent and water is 20:1:
2000, fleshing effect is preferable, and when 50:1:2000, fleshing effect is preferable, but the relatively thin position of bone and bone occur in the bones such as four limbs
Hold obscission；When the mass ratio of protease, degreasing agent and water is 30:1:2000, fleshing effect is more preferable, when 45:1:2000, goes
Meat effect is more preferable, but bone the positions such as sutura equally occurs and cracking phenomena occurs；When the mass ratio of protease, degreasing agent and water
When for 40:1:2000, fleshing effect is best.
The above embodiment is a preferred embodiment of the present invention, but embodiments of the present invention are not by above-described embodiment
Limitation, other any changes, modifications, substitutions, combinations, simplifications made without departing from the spirit and principles of the present invention,
It should be equivalent substitute mode, be included within the scope of the present invention.
Claims (10)
1. a kind of method of skeleton specimen fleshing, which comprises the following steps:
S1. the soft tissue for rejecting animal carcass 85%~90%, retains bone portion, bone is then divided into four limbs, skull, chest
Wide, lumbar vertebrae and hipbone part, encapsulate respectively；
S2. it after protease, degreasing agent and water being pressed 20~50:1~2:2000~2100 quality than mixed solution, carries out permanent
Tepidarium；
S3. the packaged bone of step S1 is respectively put into the mixed solution of step S2, so that mixed solution just floods bone
Bone impregnates 4~5 h, washes away soft tissue with high pressure water, then reject the impurity on bone；
S4. the bone of step S3 is put into the mixed solution of step S2, is dipped to bone surface free from admixture, take out bone, used
High pressure water washes away soft tissue, by bone detergent wash clean.
2. the method according to claim 1, wherein in step S2, the concentration of the protease is 0.95%~
2.5%。
3. according to the method described in claim 2, it is characterized in that, in step S2, the concentration of the protease is 1.5%~
2.5%。
4. the method according to claim 1, wherein in step S2, the concentration of the degreasing agent is 0.05%~
0.1%。
5. the method according to claim 1, wherein the protease is selected from alkali protease, acid in step S2
Property one or more of protease or neutral proteinase；It is de- that the degreasing agent is selected from alkaline defatting agent, lotion degreasing agent, solvent
One or more of rouge agent.
6. according to the method described in claim 5, it is characterized in that, the alkali protease is that Novi believes protease；The alkali
Property degreasing agent be Fur fat removal agent.
7. the method according to claim 1, wherein in step S2, the condition of the water bath with thermostatic control are as follows: in 55
~60 DEG C of 4~5 h of water bath with thermostatic control.
8. the method according to claim 1, wherein the time of immersion is 1~3 h in step S4.
9. the method according to claim 1, wherein the concentration of the detergent is 1%~2% in step S4.
10. any method according to claim 1~9, which is characterized in that four limbs, skull, thorax, lumbar vertebrae, hipbone
Total soaking time is respectively 4~8 h, 4~5 h, 4~6 h, 4~6 h, 4~6 h.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110432257A (en) * | 2019-08-01 | 2019-11-12 | 上海海洋大学 | A kind of crab class dry preserved specimen production method |
| CN111642500A (en) * | 2020-05-06 | 2020-09-11 | 广西农业职业技术学院 | Method for preparing animal bone specimen |
| CN111713486A (en) * | 2020-06-15 | 2020-09-29 | 辽宁农业职业技术学院 | Novel dog bone specimen preparation method |
| CN112598984A (en) * | 2020-12-21 | 2021-04-02 | 河北医科大学 | Fleshing device and method for manufacturing skeleton specimen |
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| CN103060406A (en) * | 2011-10-18 | 2013-04-24 | 中国科学院理化技术研究所 | Method for extracting low molecular weight collagen from cattle bone |
| CN104846048A (en) * | 2015-05-29 | 2015-08-19 | 天津市信顺康生物制品科技有限公司 | Extraction method of animal polypeptide |
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| CN111642500A (en) * | 2020-05-06 | 2020-09-11 | 广西农业职业技术学院 | Method for preparing animal bone specimen |
| CN111713486A (en) * | 2020-06-15 | 2020-09-29 | 辽宁农业职业技术学院 | Novel dog bone specimen preparation method |
| CN112598984A (en) * | 2020-12-21 | 2021-04-02 | 河北医科大学 | Fleshing device and method for manufacturing skeleton specimen |
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