CN108575825A - A kind of artificial insemination method for growing thin loach - Google Patents
A kind of artificial insemination method for growing thin loach Download PDFInfo
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- CN108575825A CN108575825A CN201810180944.XA CN201810180944A CN108575825A CN 108575825 A CN108575825 A CN 108575825A CN 201810180944 A CN201810180944 A CN 201810180944A CN 108575825 A CN108575825 A CN 108575825A
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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
- A01K61/10—Culture of aquatic animals of fish
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- 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
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
- Y02A40/81—Aquaculture, e.g. of fish
Abstract
The invention discloses a kind of artificial insemination methods for growing thin loach, include the following steps：1) physiological saline and 20ml glucose sugar juices of 20ml are extracted respectively with two 20ml syringes；2) milter sperm is drawn with dry suction pipe；3) first brine is squeezed into a plastic tub when artificial insemination, then sperm is squeezed into the brine in plastic tub, by semen dilution；4) raun is taken in one's arms and dries moisture, fish-egg is extruded on diluted sperm, being gently mixed with feather makes smart ovum be sufficiently mixed；5) ready glucose solution is squeezed into again after mixing and activates sperm in basin；6) 23 completion fertilization processes of clear water cleaning fish-egg are added after 12 minutes；7) finally fertilized eggs are transferred in incubator and are hatched；The present invention mainly uses short-term holding of the physiological saline to sperm, has certain help to the nonsynchronous parent population of male and female, the parent population few to milter essence amount, and physiological saline can be utilized to preserve and dilute, and provides nutrition using glucose, greatly improves the rate of fertilization of fish-egg.
Description
Technical field
The invention belongs to technical field of aquaculture, specifically, being related to a kind of artificial insemination method for growing thin loach.
Background technology
Long thin loach (Leptobotia elongata, Bleeker) also known as flower loach, thin loach, belong to Cobitidae, Botiinae, thin loach
Category is the distinctive type in China, is distributed mainly on upper Changjiang region middle and upper reaches and its tributary, and in Cobitidae fish, individual is maximum, grows
Most fast, general individual 0.2-0.4kg, maximum individual reaches 3.0kg.Long thin loach is bright in colour, head and trunk body surface point
It is furnished with irregular dark brown speckle, ornamental value is high, enjoys high reputation in the domestic and international markets, and 1989 in state of Singapore
Gold medal is won in the ornamental fish comparation and assessment of border, it is a kind of simultaneous ornamental and edible famous-brand and high-quality economic fish, and there is fabulous market to open
Hair foreground.National endangered species Science committees in 1998 and State Bureau of Environmental Protection are included in Chinese animals on the brink of extinction Red Data Book (Yin
Red rosy clouds, 2006).
However since overfishing, the construction of hydropower stations are to caused by the destruction in the thin loach spawning ground of length and the fast development of industry
The influence of the factors such as water pollution so that the wild resource of long thin loach drastically declines, therefore long thin loach species conservation work is urgently
Carry out.Artificial propagation of fish and enhancement releasing are one of current development species conservation, the increase most effective measure of population quantity.It is complete
Kind parent population artificial propagation techniques are to ensure the Technique in Fishes enhancement releasing prerequisite that everything goes well with your work carries out.Recent year
Person is many for the thin loach correlative study of length, is related to biology, artificial propagation, seed rearing etc..The long thin loach people in these researchs
Work breed and seed rearing be carried out under pond condition (Zhao Yunfang, 1995；Liang Yinquan etc., 1999；Liang Yinquan etc.,
2007；Liang Yinquan etc., 2000；Sun great Dong etc., 2010), but the research under fish recirculating aquaculture system condition indoors is not yet
It appears in the newspapers.
The artificial propagation for growing thin loach at present is also in a primary research stage, and the parent population maturing rate that the interior pool is cultivated is low, special
Be not milter essence amount it is few, and male and female are mostly asynchronous, and conventional dry method Insemination procedures are first to squeeze sperm after crowded ovum, often
Sperm is difficult to squeeze out or seldom, seriously affect rate of fertilization when ovum squeezes sperm again after squeezing out.
To solve the above problems, the present invention provides a kind of artificial insemination method for growing thin loach, this method is simple, operation side
Just, overcome above deficiency, can be effectively improved in the case where poor, asynchronous, the smart amount of parent population maturing rate is few long thin loach by
Smart rate.
Invention content
The present invention is directed to the deficiencies in the prior art, it is therefore an objective to a kind of artificial insemination method for growing thin loach is provided, it should
Milter sperm is carried out short-term preservation by method in the breeding of the thin loach of length in physiological saline, is utilized when raun smoothly ovulates
The glucose of amount is activated with water (salinity 0.45%), ensure that the vigor of sperm, improves the rate of fertilization of fish-egg, and simple,
It is easy to operate.
To achieve the above object, the technical solution used in the present invention is：
A kind of artificial insemination method for growing thin loach, includes the following steps：
(1) preparation of reagent：
A, prepare the physiological saline and each one bottle of glucose injection of certain mass concentration respectively, and put it into and hasten parturition
It is impregnated in the consistent water body of water temperature, it is spare；
B, prepare one bag of MS222 anesthetic, it is spare；
(2) preparation of tool：
A, prepare plastic tub 1-2 of inner wall smooth, fish presss from both sides 2, the plastic suction pipe 2-5 that 20ml syringes 2-4 is a, dry
A, towel 2-4 items, square plastic magazine 2, feather 3-5 roots；
B, by plastic tub towel moisture；2 plastic casings are loaded onto into the clear water consistent with fish spawning inducing pool water temperature, are pressed
MS222 anesthetic is added in a plastic casing thereto for the concentration of 50-100ppm, and drug is uniformly mixed spare；
C, prepare an oxygen increasing pump；
(3) artificial insemination operates：
A, oxytocic drug will be injected to reach Effect time and start the thin loach parent population seine concentration of male and female length that heat is ovulated,
Then the male and female parent population of smart ovum can smoothly be squeezed out by, which selecting, is put into the plastic casing for fill anesthetic solution and anaesthetizes；
B, it takes the syringe of a 20mL to draw 20ml physiological saline from the normal saline solution for regulate water temperature to squeeze
Into ready plastic tub；Glucose injection with another 20ml syringe draws equal amounts is spare；
C, a tail milter of holonarcosis is taken to be put into fish folder, abdomen holds out the water surface upward, and fish body anus is dried with dry towel
The moisture at each position with door, fish folder and operator, ensures that the water containing anesthetic not mix in sperm and drips in basin, use hand
Light pressure fish belly portion squeezes out sperm, is used in combination plastic suction pipe that milter sperm is drawn onto in plastic tube, is then again extruded into sperm and fills life
In the plastic tub for managing brine, parent population is put into the plastic casing that another fills clear water makes it slowly revive later；Then one is selected
Tail raun is put into fish folder, and parent population is wrapped, covers anus on the other hand, takes fish folder to embrace by parent population on the other hand, fish is dried with dry towel
The moisture at each position, ensures that the water containing anesthetic not drip in basin, fish belly portion is gently pressed with hand with body, fish folder and operator
Ovum is squeezed out, ovum is expressed into the plastic tub equipped with physiological saline and sperm, and parent population is also put into the modeling for filling clear water later
In magazine and opening oxygen increasing pump oxygenation makes it slowly revive；Then, the physiology salt containing sperm and ovum is gently agitated for feather
Water 5-20 seconds makes it be sufficiently mixed uniformly, then squeezes into the glucose injection of 20ml on the smart ovum in basin, then light with feather
Light stirring makes sperm fully activate and makes ovum fertilization for 20-30 seconds；
D, 1-2kg clear water is added into the above-mentioned plastic tub for filling sperm and ovum, stirring 10-20 seconds or so is rear to stand 1
Minute or so, finally fertilized eggs are cleaned with a large amount of clear water 2-3 times, finally to being hatched in incubator.
Preferably, the mass concentration of the physiological saline is 0.9%, and the mass concentration of glucose injection is 5%；The two
Volume be 250-500mL.
Preferably, the charge weight of clear water is about the 1/2 of plastic casing volume in the plastic casing.
Preferably, the capacity that the square plastic magazine is filled with water is more than 50kg.
Preferably, the amount of squeezing into of sperm is 2-3 drops in plastic tub.
Artificial insemination method of the present invention mainly substantially conforms to the salinity of parent population body fluid by the salinity of physiological saline, can make essence
Liquid energy is preferably preserved in a short time, and glucose can be activated and be provided nutrient energy, improves sperm motility,
Achieve the purpose that higher rate of fertilization.
Compared with prior art, the present invention has the following advantages and beneficial effect：
1) convenient material drawing of the present invention, of low cost, operating method is simple；
2) present invention is to use short-term holding of the physiological saline to sperm, the parent population few to milter essence amount that can utilize physiology
Brine preserves, and provides nutrient energy using glucose, greatly improves the rate of fertilization of fish-egg, it is different to efficiently solve parent population male and female
The problem of step；
3) present invention is diluted milter sperm using physiological saline, so that smart ovum is uniformly mixed rapidly, reaches fully close
Distance contacts and provides nutrition and energy using glucose for sperm, improves vigor and the service life of sperm, effectively improves fish-egg
Rate of fertilization.
Specific implementation mode：
Technical scheme of the present invention is described in further detail below in conjunction with specific embodiments.It should be understood that institute
Purpose for embodiment is the content that the present invention is further explained, and cannot be construed to protect the present invention in any sense
The limitation of range.
Embodiment 1：
On July 11st, 2017, at Sichuan Province Dadu River domestic taking food as usual fish multiplication station, circulation is hastened parturition hatching and seedling
1 mouthful of 2m of kind cultivated workshop, which is cultivated, selects 6 groups of (sex rations 2 of ripe preferably long thin loach parent population in cylinder:2) it is manually urged
Production is divided to two needles to inject, and the first needle time was the morning 18:When 00, the second needle is evening 24:When 00, water temperature of hastening parturition is 22 DEG C.To
Two days 11:It finds that parent population has the sign for chasing heat when 00, but chases unobvious.To 12：It is apparent sharp to find that parent population chases when 00
It is strong, find that parent population (raun) has started to ovulate when parent population is picked up inspection, it is light that abdomen is pressed to have ovum outflow, milter that can also squeeze out on a small quantity
Sperm is ready for artificial insemination.This time two methods of artificial insemination point carry out, and are respectively 3 groups of parent populations, and one group is test group,
Using first squeeze sperm to physiological saline in, then afterwards squeeze ovum to basin in, finally plus the diluted method of glucose be fertilized；Control group
It is fertilized by our common dry method insemination methods.It is specific as follows：
One, test group
The artificial insemination method of long thin loach, includes the following steps：
(1) preparation of reagent：
A, prepare that 250ml fills 0.9% physiological saline and 250ml fills each one bottle of 5% glucose injection respectively, and by its
It is put into the water body consistent with water temperature of hastening parturition and impregnates, it is spare；
B, it is spare to prepare one bag of MS222 anesthetic；
(2) preparation of tool：
A, the plastic tub 1 of preparation inner wall smooth, 2, fish folder, 20ml syringes 2, dry plastic suction pipe 5, mao
4, towel, the square plastic magazine 2 of 70kg capacity and 5, feather；
B, by plastic tub towel moisture；2 plastic casings are loaded onto into the clear water consistent with fish spawning inducing pool water temperature, water
MS222 anesthetic is added in a plastic casing 1/2 or so of plastic casing total measurement (volume), and by the concentration of 50ppm thereto for control,
Then drug is uniformly mixed spare；
C, prepare an oxygen increasing pump, including power supply, wiring board and related oxygen pipe, gas stone；
(3) artificial insemination operates：
A, the male and female parent population (1 tail raun, 1 tail milter) of smart ovum can smoothly be squeezed out by, which selecting, is put into the modeling for filling anesthetic solution
It is anaesthetized in magazine；
B, the syringe of a 20mL is taken to draw 20ml physiology from the physiological saline that (1) the step of regulating water temperature prepares
Brine is extruded into ready plastic tub, and the glucose injection with another 20ml syringe draws equal amounts is spare；
C, a tail milter of holonarcosis is taken to be put into fish folder, anus dries fish body upward, with dry towel, fish is pressed from both sides and operation
The moisture (ensureing that the water containing anesthetic not drip in basin) at each position, gently presses fish belly portion to squeeze out sperm with hand on the person, and
Milter sperm is drawn onto in plastic tube with plastic suction pipe, sperm is then extruded into the plastic tub for filling physiological saline (3 drop of drop again
Sperm), being later put into parent population in another plastic casing for fill clear water makes it slowly revive；Then 1 tail raun is selected to be put into fish folder
In wrap, cover anus (ensuring that fish-egg not flow out arbitrarily) on the other hand, catch on the other hand fish folder by parent population embrace, dried with dry towel
The moisture (ensureing that the water containing anesthetic not drip in basin) at each position, fish is gently pressed with hand with fish body, fish folder and operator
Abdomen squeezes out ovum, and ovum is expressed into the plastic tub equipped with physiological saline and sperm, is also put into parent population fills clear water later
Plastic casing in and opening oxygen increasing pump oxygenation makes it slowly revive；Then, the life containing sperm and ovum is gently agitated for feather
It manages brine 20 seconds, it is made to be sufficiently mixed uniformly, and the glucose of 20ml is squeezed into water in basin on smart ovum, then gently stirred with feather
Mixing 30 seconds makes sperm fully activate and makes ovum fertilization；
D, 1.5kg clear water, the rear standings of stirring 10s or so is added after 1min into the above-mentioned plastic tub for filling sperm and ovum
1 minute or so, finally fertilized eggs are cleaned with a large amount of clear water 3 times, then obtain the (one kind at present of fertilized eggs 0.8 ten thousand with graduated cylinder measurement Law
Common metering method), and move on in a self-control hatching basket and carry out Lotic hatching, hatching water temperature is 22.1 DEG C；
E, the artificial insemination that remaining 2 groups of parent populations are carried out with same method works.Finally 3 groups of parent populations of metering obtain fertilized eggs altogether
About 2.6 ten thousand.
(4) rate of fertilization counts：
By hatching in 10 hours, fertilized eggs developed primitive gut mid-term.At random with fry dredger fish for a small amount of fish-egg into
Row rate of fertilization counts (fish for be averaged three times at random).Statistical result is：128, ovum, fertilized eggs 109 are taken for the first time, by
Smart rate 85.16%；136, ovum, fertilized eggs 116, rate of fertilization 85.29% are taken for the second time；Third time takes 187, ovum, fertilized eggs
159, rate of fertilization 85.03%；Average rate of fertilization 85.14%.
Two, control group
Dry method of artificial fertilization's method of long thin loach, includes the following steps：
1, the reagent and tool of prefecundation prepares
A, prepare one bag of MS222 anesthetic, the plastic tub 1 of inner wall smooth, 2, fish folder, 20ml syringes 2, drying
Plastic suction pipe 2, towel 4,2,5 feathers of 70kg square plastics magazine；
B, by plastic tub towel moisture；2 plastic casings are loaded onto into the clear water consistent with fish spawning inducing pool water temperature, water
MS222 anesthetic is added in a plastic casing 1/2 or so of plastic tub total measurement (volume), and by the concentration of 50ppm thereto for control,
And drug is uniformly mixed spare；And prepare an oxygen increasing pump, including power supply, wiring board and related oxygen pipe, gas stone；
2, artificial insemination operations
A, the male and female parent population (1 tail raun, 1 tail milter) that will smoothly squeeze out smart ovum is put into fiber crops in the solution for fill anesthetic
It is liquor-saturated；
B, after parent population is anesthetized completely, 1 tail raun is put into during fish is pressed from both sides and proposes by a people, dries fish body with dry towel, fish is pressed from both sides
With the moisture (ensureing that the water containing anesthetic not drip in basin) at each position with operator, ovum is extruded into ready wiping
In the basin of solid carbon dioxide point, parent population, which has squeezed after ovum to be put into another and fill in the plastic casing of clear water, makes it slowly revive；Then by milter
Sperm, which is extruded into the plastic tub equipped with ovum, with same method (be put into fish folder, dry moisture, crowded essence) (can only squeeze out 3-4
Drip sperm), then 15 seconds (smart ovum is made to be sufficiently mixed) is gently agitated for feather；It is subsequently added into a small amount of clear water and is gently stirred with feather
Mixing 25 seconds makes sperm fully activate and makes ovum fertilization；
C, fertilized eggs are cleaned 3 times with a large amount of clear water after 1 minute, then 0.6 ten thousand, ovum is obtained with graduated cylinder metering, move on to a self-control
Lotic hatching is carried out in hatching basket, hatching water temperature is 22.1 DEG C.
D, the artificial insemination operations of other 2 groups of parent populations are carried out with same method.It finally counts and obtains 2.4 ten thousand left sides of ovum altogether
It is right.
3, rate of fertilization counts
By hatching in 10 hours, fertilized eggs developed primitive gut mid-term.At random with fry dredger fish for a small amount of fish-egg into
Row rate of fertilization counts (fish for be averaged three times at random).Statistical result is：115, ovum, fertilized eggs 33 are taken for the first time, are fertilized
Rate 28,7%；177, ovum, fertilized eggs 54, rate of fertilization 30.5% are taken for the second time；Third time takes 254, ovum, fertilized eggs 77,
Rate of fertilization 30.3%, average rate of fertilization 30.04%.
Embodiment 2
On June 14th, 2017, the domestic Jinsha jiang River middle reaches Datang kwan-yin rock power station fish multiplication of Panzhihua City in Sichuan Province
4 groups of (sex rations 1 of mature brood fish are selected in pouring station outdoor tank-culture:1) it hastens parturition.It is divided to two needles to inject, the first needle time
For the morning 9:When 00, the second needle is evening 15:When 30, water temperature of hastening parturition is 24 DEG C.To 22:00 estimates Effect time pull-up net cage
It carries out observation and finds that parent population does not ovulate also.Find that parent population chases heat phenomenon in Chi Zhongyou after half an hour, when picking up parent population inspection
It was found that raun has started to ovulate, it is light that abdomen is pressed to have ovum outflow, milter that can also squeeze out a small amount of sperm, that is, be ready for artificial insemination.
This time two methods of artificial insemination point carry out, and one kind is test group, and 2 groups of parent population, use is selected first to squeeze in sperm to physiological saline,
It squeezes in ovum to basin afterwards again, finally plus the diluted method of glucose is fertilized；2 groups of control group parent population, by our common dry method
Insemination method is fertilized.It is specific as follows：
One, test group
The artificial insemination method of long thin loach, step are：
(1) preparation of reagent：
A, prepare that 250ml fills 0.9% physiological saline and 250ml fills each one bottle of 5% glucose injection respectively, and by its
It is put into the water body consistent with water temperature of hastening parturition and impregnates, it is spare；
B, it is spare to prepare one bag of MS222 anesthetic；
(2) preparation of tool：
A, the plastic tub 1 of preparation inner wall smooth, 2, fish folder, 20ml syringes 2, dry plastic suction pipe 4, mao
4, towel, 70kg square plastics magazine 2 and 5, feather；
B, by plastic tub towel moisture；2 plastic casings are loaded onto into the clear water consistent with fish spawning inducing pool water temperature, water
MS222 anesthetic is added in a plastic casing 1/2 or so of plastic tub total measurement (volume), and by the concentration of 50ppm thereto for control,
Then drug is uniformly mixed spare；
C, prepare an oxygen increasing pump, including power supply, wiring board and related oxygen pipe, gas stone；
(3) artificial insemination operates：
A, the male and female parent population (1 tail raun, 1 tail milter) of smart ovum can smoothly be squeezed out by, which selecting, is put into the modeling for filling anesthetic solution
It is anaesthetized in magazine；
B, the syringe of a 20mL is taken to draw 20ml lifes in the physiological saline of preparation from (1) the step of regulating water temperature
Reason brine is extruded into ready plastic tub, and the glucose injection with another 20ml syringe draws equal amounts is spare；
C, a tail milter of holonarcosis is taken to be put into fish folder, abdomen dries fish body upward, with dry towel, fish is pressed from both sides and operation
The moisture (ensureing that the water containing anesthetic not drip in basin) at each position, gently presses fish belly portion to squeeze out sperm with hand on the person, and
Milter sperm is drawn onto in plastic tube with plastic suction pipe, then sperm is extruded into the plastic tub for filling physiological saline again, later
Parent population, which is put into the plastic casing that another fills clear water, makes it slowly revive；Then it selects 1 tail raun to be put into successively in fish folder to wrap
It is good, anus (ensuring that fish-egg not flow out arbitrarily) is covered on the other hand, catches fish folder to embrace by parent population on the other hand, fish is dried with dry towel
The moisture (ensureing that the water containing anesthetic not drip in basin) at each position, fish belly is gently pressed with hand with body, fish folder and operator
Portion squeezes out ovum, and ovum is expressed into the plastic tub equipped with physiological saline and sperm, is also put into parent population fills clear water later
In plastic casing and opening oxygen increasing pump oxygenation makes it slowly revive；Then, the physiology containing sperm and ovum is gently agitated for feather
Brine 20 seconds makes it be sufficiently mixed uniformly, and the glucose of 20ml is squeezed into water in basin on smart ovum, then is gently mixed with feather
25 are uniformly mixed, and so that sperm is fully activated and make ovum fertilization；
D, 1kg clear water is added after 30 seconds into the above-mentioned plastic tub for filling sperm and ovum, stirring 10s or so is rear to stand 1
Minute, finally fertilized eggs are cleaned with a large amount of clear water 2 times, then be put on balance and weighed, it is 0.6 ten thousand to calculate gained fish-egg,
Then it is moved into self-control hatching basket and carries out Lotic hatching, hatching water temperature is 22.5 DEG C.
E, same method is taken to carry out the artificial insemination of second group of parent population, counting obtains 0.75 ten thousand, ovum.
(4) rate of fertilization counts
By hatching in 8 hours, development of fertilized ova to primitive gut mid-term.At random with fry dredger fish for a small amount of fish-egg carry out by
Smart rate statistics (2 groups each random fishing sample 3 times be averaged).
Statistical result is：
First group takes 115, ovum, wherein fertilized eggs 98, rate of fertilization 85.22% for the first time；Take for the second time 318, ovum, by
270, smart ovum, rate of fertilization 84.91%；Third time takes 284, ovum, fertilized eggs 242, rate of fertilization 85.21%, average value
85.08%.
Second group takes 166, ovum, wherein fertilized eggs 141, rate of fertilization 84.94% for the first time；Take for the second time 189, ovum,
Wherein fertilized eggs 162, rate of fertilization 85.71%；Third time takes 336, ovum, wherein fertilized eggs 285, rate of fertilization 84.82%,
Average out to 85.09%.
Two groups of average rate of fertilization is 85.085%.
Two, control group
Dry method of artificial fertilization's method of long thin loach, step are：
1, the reagent and tool of prefecundation prepares：
A, prepare one bag of MS222 anesthetic, the plastic tub 2 of inner wall smooth, 2, fish folder, 20ml syringes 2, towel 4
2,5 feathers of square plastic magazine of item, capacity 70kg；
B, by plastic tub towel moisture；2 plastic casings are loaded onto into the clear water consistent with fish spawning inducing pool water temperature, water
Control, by MS222 is added in 50ppm concentration thereto a plastic casing, drug is mixed 1/2 or so of plastic casing total measurement (volume)
It closes uniformly spare；Prepare an oxygen increasing pump, including power supply, wiring board and related oxygen pipe, gas stone.
2, artificial insemination operations：
A, 1 group of the male and female parent population (female each 1 tail of milter) of smart ovum can smoothly be squeezed out by, which selecting, is put into the solution for filling anesthetic
Anesthesia；
B, after parent population is anesthetized completely, 1 tail raun is put into during fish is pressed from both sides and proposes by a people, dries fish body with dry towel, fish is pressed from both sides
With the moisture (ensureing that the water containing anesthetic not drip in basin) at each position with operator, ovum is extruded into ready wiping
In the plastic tub of solid carbon dioxide point.Milter is used the above method (be put into fish folder, dry moisture, crowded essence) by 1 tail hero by another people simultaneously
The sperm of fish is extruded into the plastic tub equipped with ovum, is then gently agitated for 20 seconds with feather, and a small amount of clear water is added later and uses feather
Being gently mixed 30s activation sperms makes ovum fertilization；
C, fertilized eggs are cleaned 2 times with a large amount of clear water after 1min, then 0.65 ten thousand, ovum is obtained altogether with balance weighing apparatus weighing, it will after weighing
Fish-egg moves on in another self-control hatching basket and carries out recirculated water hatching, and hatching water temperature is 22.5 DEG C；
D, the artificial insemination for then carrying out other 1 group of parent population in aforementioned manners, obtains 1.2 ten thousand, ovum, and moves on to other incubate
Change and hatch in basket, it is also 22.5 DEG C to hatch water temperature to be also.
3, rate of fertilization counts
By 8 hours hatching development of fertilized ova to primitive gut mid-term.At random with fry dredger fish for a small amount of fish-egg carry out by
Smart rate statistics (2 groups each random fish for 3 times be averaged).
Statistical result is：
1st group：Take 145, ovum, fertilized eggs 43, rate of fertilization 29.66% for the first time；248, ovum, fertilized eggs are taken for the second time
74, rate of fertilization 29.84%；Third time takes 176, ovum, fertilized eggs 53, rate of fertilization 30.11%, average value 29.88%；
Second group：Take 214, ovum, fertilized eggs 64, rate of fertilization 29.9% for the first time；237, ovum, fertilized eggs are taken for the second time
72, rate of fertilization 30.38%；Third time takes 302, ovum, fertilized eggs 91, rate of fertilization 30.13%, average value 30.15%；
Two groups of average rates of fertilization are 30.02%.
It should be pointed out that specific implementation mode described above can make those skilled in the art that this hair be more fully understood
It is bright, but do not limit the invention in any way.Therefore, it will be appreciated by those skilled in the art that still can be carried out to the present invention
Modification or equivalent replacement；And technical solution and its improvement of all spirit and technical spirit that do not depart from the present invention, it should all
Cover in the protection domain of patent of the present invention.
Claims (5)
1. a kind of artificial insemination method for growing thin loach, which is characterized in that include the following steps：
(1) preparation of reagent：
A, prepare the physiological saline and each one bottle of glucose injection of certain mass concentration, and the water temperature that puts it into and hasten parturition respectively
It is impregnated in consistent water body, it is spare；
B, prepare one bag of MS222 anesthetic, it is spare；
(2) preparation of tool：
A, the plastic tub of preparation inner wall smooth 1-2,2, fish folder, 20ml syringes 2-4 are a, drying plastic suction pipe 2-5 is a,
Towel 2-4 items, square plastic magazine 2, feather 3-5 roots；
B, by plastic tub towel moisture；2 plastic casings are loaded onto into the clear water consistent with fish spawning inducing pool water temperature, by 50-
MS222 anesthetic is added in a plastic casing thereto for the concentration of 100ppm, is then uniformly mixed drug spare；
C, prepare an oxygen increasing pump；
(3) artificial insemination operates：
A, oxytocic drug will be injected to reach Effect time and start the thin loach parent population seine concentration of male and female length that heat is ovulated, then
The male and female parent population of smart ovum can smoothly be squeezed out by, which selecting, is put into the plastic casing for fill anesthetic solution and anaesthetizes；
B, it takes the syringe of a 20mL to draw 20ml physiological saline from the normal saline solution for regulate water temperature and is extruded into standard
In the plastic tub got ready；Glucose injection with another 20ml syringe draws equal amounts is spare；
C, a tail milter of holonarcosis is taken to be put into fish folder, abdomen exposes the surface upward, and fish body anus, fish are dried with dry towel
The moisture of folder and each position with operator ensures that the water containing anesthetic not mix in sperm and drips in basin, gently pressed with hand
Fish belly portion squeezes out sperm, is used in combination plastic suction pipe that milter sperm is drawn onto in plastic tube, is then again extruded into sperm and fills physiology salt
In the plastic tub of water, parent population is put into the plastic casing that another fills clear water makes it slowly revive later；Then select a tail female
Fish is put into fish folder, covers anus on the other hand, is pressed from both sides with fish embrace the water surface by parent population on the other hand, dries fish body with dry towel, fish is pressed from both sides and behaviour
The moisture for making the upper each position of the person ensures that the water containing anesthetic not drip in basin, fish belly portion is gently pressed to squeeze out ovum, ovum with hand
Son is expressed into the plastic tub equipped with physiological saline and sperm, is later also put into parent population in the plastic casing for filling clear water and is opened
Oxygen increasing pump oxygenation makes it slowly revive；Finally, it is gently agitated for the physiological saline containing sperm and ovum 5-20 seconds with feather, makes it
It is sufficiently mixed uniformly, then squeezes into the glucose injection of 20ml on the smart ovum in basin, then be gently mixed 20-30 seconds with feather
So that sperm is fully activated and makes ovum fertilization；
D, 1-2kg clear water is added into the above-mentioned plastic tub for filling sperm and ovum, stirring 10-20 seconds or so is rear to stand 1 minute
Left and right, then clean fertilized eggs 2-3 times with a large amount of clear water, it then moves on in incubator and is hatched.
2. Insemination procedures according to claim 1, which is characterized in that the mass concentration of the physiological saline is 0.9%, Portugal
The mass concentration of grape sugar injection is 5%；The volume of the two is 250-500mL.
3. according to claim 1-2 any one of them Insemination procedures, which is characterized in that the charge weight of clear water in the plastic casing
About the 1/2 of plastic casing volume.
4. according to claim 1-3 any one of them Insemination procedures, which is characterized in that the capacity that the square plastic magazine is filled with water
More than 50kg.
5. according to claim 1-4 any one of them Insemination procedures, which is characterized in that the amount of squeezing into of sperm is 2- in plastic tub
3 drops.
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