CN109122668A - A kind of store method of hilsa herring sperm - Google Patents
A kind of store method of hilsa herring sperm Download PDFInfo
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- CN109122668A CN109122668A CN201811249646.8A CN201811249646A CN109122668A CN 109122668 A CN109122668 A CN 109122668A CN 201811249646 A CN201811249646 A CN 201811249646A CN 109122668 A CN109122668 A CN 109122668A
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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
- A01N1/02—Preservation of living parts
- A01N1/0205—Chemical aspects
- A01N1/021—Preservation or perfusion media, liquids, solids or gases used in the preservation of cells, tissue, organs or bodily fluids
- A01N1/0226—Physiologically active agents, i.e. substances affecting physiological processes of cells and tissue to be preserved, e.g. anti-oxidants or nutrients
Abstract
The invention discloses a kind of store methods of hilsa herring sperm, after the 1:1:5 mixing by volume of antifreeze, hilsa herring sperm and dilution, and save at 5 DEG C；Dilution volume percent composition are as follows: sodium chloride injection: 50%, pure water: 15-20%, potassium chloride: 7-14%, calcium chloride: 5-10%, sodium bicarbonate: 5-10%, chlorogenic acid: 1-2%, glycerol: 1-3%, glucose: 3-5%, lecithin: 8-15%, sodium citrate: 1-3%, streptomysin: 1-2%, polyglutamic acid sodium: 1-3%, Bifidobacterium: 1-3%, surplus are lactic acid bacteria；Antifreeze percent by volume ingredient are as follows: dimethyl sulfoxide: 30%, glycerol: 20%, ethylene glycol: 30%, dimethyl acetamide: 20%；The present invention not only effectively inhibits harmful microbe growth and breeding, but also reduces sperm metabolism rate, effectively maintains the preservation quality of hilsa herring sperm, has saved production cost, and operating procedure is convenient.
Description
Technical field
The present invention relates to aquatic reproduction technology fields, particularly relate to a kind of store method of hilsa herring sperm.
Background technique
Hilsa herring is anadromous migration fishes, and because entering river when annual timing early summer, other times do not occur, therefore
Very precious, so people extremely value the breeding of hilsa herring, there are mainly two types of the sperm that hilsa herring artificial insemination uses, i.e., fresh
Sperm and frozen semen, the former is often simply called fresh essence, and the latter referred to as freezes essence.For the utilization rate for improving kind of hilsa herring, it is convenient for sperm
Preservation and transport, some kinds of hilsa herring farms often make jelly essence, the breeding work for hilsa herring.But the equipment that essence is frozen in production is high
It is expensive, it saves transportational process and needs to be replenished in time liquid nitrogen, semen thawing technical requirements are higher, and the rate of fertilization that hilsa herring freezes essence is more low,
These disadvantages for freezing essence for hilsa herring, are difficult popularization and application to each farm and raiser；Thus greatly limited using jelly essence
The popularization and application of hilsa herring estrus synchronization and artificial insemination the relevant technologies；
Production line is usually diluted using the multi-purpose physiological saline of fresh essence, and this method only can satisfy live semen collection, scene
Dilution, the production requirement of instant semen deposition；Currently, possessing a large amount of hilsa herring extender of freezing semen and dilution preservation liquid in the market；Fresh essence
Dilution and dilution save that liquid is less, and Main Components are mostly glucose, sodium citrate, antibiotic, sperm motility rate, and plasma membrane is complete
Whole rate, acrosomal integrity is ideal not to the utmost, and all reduces significantly after 48 hours.
Summary of the invention
The technical problem to be solved by the present invention is to propose a kind of hilsa herring sperm for the above disadvantage of the existing technology
Store method, not only effectively inhibit harmful microbe growth and breeding, but also reduce sperm metabolism rate, it is effective to keep
The preservation quality of hilsa herring sperm, has saved production cost, and operating procedure is convenient.
The technical solution that the present invention solves the above technical problem is:
A kind of store method of hilsa herring sperm, after the 1:1:5 mixing by volume of antifreeze, hilsa herring sperm and dilution, and
5 DEG C of preservations；Dilution volume percent composition are as follows: sodium chloride injection: 50%, pure water: 15-20%, potassium chloride: 7-14%, chlorination
Calcium: 5-10%, sodium bicarbonate: 5-10%, chlorogenic acid: 1-2%, glycerol: 1-3%, glucose: 3-5%, lecithin: 8-15%, citric acid
Sodium: 1-3%, streptomysin: 1-2%, polyglutamic acid sodium: 1-3%, Bifidobacterium: 1-3%, surplus are lactic acid bacteria；Antifreeze volume basis
Compare ingredient are as follows: dimethyl sulfoxide: 30%, glycerol: 20%, ethylene glycol: 30%, dimethyl acetamide: 20%.
The technical solution that the present invention further limits is:
The store method of hilsa herring sperm above-mentioned, dilution volume percent composition are as follows: sodium chloride injection: 50%, pure water:
15%, potassium chloride: 7%, calcium chloride: 5%, sodium bicarbonate: 5%, chlorogenic acid: 1%, glycerol: 1%, glucose: 3%, lecithin: 8%, lemon
Lemon acid sodium: 1%, streptomysin: 1%, polyglutamic acid sodium: 1%, Bifidobacterium: 1%, surplus is lactic acid bacteria.
The store method of hilsa herring sperm above-mentioned, dilution volume percent composition are as follows: sodium chloride injection: 50%, it is pure
Water: 20%, potassium chloride: 14%, calcium chloride: 10%, sodium bicarbonate: 10%, chlorogenic acid: 2%, glycerol: 3%, glucose: 5%, lecithin:
15%, sodium citrate: 3%, streptomysin: 2%, polyglutamic acid sodium: 3%, Bifidobacterium: 3%, surplus is lactic acid bacteria.
The store method of hilsa herring sperm above-mentioned, dilution volume percent composition are as follows: sodium chloride injection: 50%, it is pure
Water: 18%, potassium chloride: 12%, calcium chloride: 8%, sodium bicarbonate: 8%, chlorogenic acid: 1%, glycerol: 1%, glucose: 3%, lecithin:
10%, sodium citrate: 2%, streptomysin: 1%, polyglutamic acid sodium: 2%, Bifidobacterium: 2%, surplus is lactic acid bacteria.
The store method of hilsa herring sperm above-mentioned, the preparation of dilution the following steps are included:
(1) 100ml pure water is measured, in 100-110 DEG C of high pressure sterilization 10-12min, obtains sterilizing pure water A；
(2) lemon sodium bicarbonate, glucose, sodium citrate, Bifidobacterium, streptomysin and lactic acid bacteria are weighed, is added separately to go out
In bacterium pure water A, in 55-65 DEG C of heating water bath, and stirring and dissolving 10-15min clockwise is carried out, then gone out in 120 DEG C of high pressures
Bacterium 15-20min obtains solution B；
(3) the sterile fresh yolk liquid of absorption is added in the centrifuge tube of sterilizing after carrying out 1:1 dilution with sterile saline,
It is centrifuged 10-15min in 5 DEG C of 7500g, draws sodium chloride injection 10-15ml, in 15-25 DEG C of standing 8-12min, obtains ovum
Yellow centrifuged supernatant C；
(4) potassium chloride and calcium chloride are weighed, is added separately in yolk centrifuged supernatant C, stirs and evenly mixs clockwise, is obtained
Solution D；
(5) it takes out solution B and weighs chlorogenic acid, glycerol, lecithin and polyglutamic acid when its temperature drops to 85 DEG C or less
Sodium is added in solution B, and carries out stirring and dissolving clockwise, obtains solution E；
(6) when solution E temperature drops to 35-40 DEG C, solution D is slowly poured into, mixing.
The store method of hilsa herring sperm above-mentioned, the preparation of dilution the following steps are included:
(1) 100ml pure water is measured, in 100 DEG C of high pressure sterilization 10min, obtains sterilizing pure water A；
(2) lemon sodium bicarbonate, glucose, sodium citrate, Bifidobacterium, streptomysin and lactic acid bacteria are weighed, is added separately to go out
In bacterium pure water A, in 55 DEG C of heating water baths, and stirring and dissolving 10min clockwise is carried out, then in 120 DEG C of high pressure sterilizations
15min obtains solution B；
(3) the sterile fresh yolk liquid of absorption is added in the centrifuge tube of sterilizing after carrying out 1:1 dilution with sterile saline,
It is centrifuged 10min in 5 DEG C of 7500g, draws sodium chloride injection 10ml, in 15 DEG C of standing 8min, obtains yolk centrifuged supernatant
C；
(4) potassium chloride and calcium chloride are weighed, is added separately in yolk centrifuged supernatant C, stirs and evenly mixs clockwise, is obtained
Solution D；
(5) it takes out solution B and weighs chlorogenic acid, glycerol, lecithin and polyglutamic acid when its temperature drops to 85 DEG C or less
Sodium is added in solution B, and carries out stirring and dissolving clockwise, obtains solution E；
(6) when solution E temperature drops to 35 DEG C, solution D is slowly poured into, mixing.
The store method of hilsa herring sperm above-mentioned, the preparation of dilution the following steps are included:
(1) 100ml pure water is measured, in 110 DEG C of high pressure sterilization 12min, obtains sterilizing pure water A；
(2) lemon sodium bicarbonate, glucose, sodium citrate, Bifidobacterium, streptomysin and lactic acid bacteria are weighed, is added separately to go out
In bacterium pure water A, in 65 DEG C of heating water baths, and stirring and dissolving 15min clockwise is carried out, then in 120 DEG C of high pressure sterilizations
20min obtains solution B；
(3) the sterile fresh yolk liquid of absorption is added in the centrifuge tube of sterilizing after carrying out 1:1 dilution with sterile saline,
It is centrifuged 15min in 5 DEG C of 7500g, draws sodium chloride injection 15ml, in 25 DEG C of standing 12min, yolk is obtained and is centrifuged supernatant
Liquid C；
(4) potassium chloride and calcium chloride are weighed, is added separately in yolk centrifuged supernatant C, stirs and evenly mixs clockwise, is obtained
Solution D；
(5) it takes out solution B and weighs chlorogenic acid, glycerol, lecithin and polyglutamic acid when its temperature drops to 85 DEG C or less
Sodium is added in solution B, and carries out stirring and dissolving clockwise, obtains solution E；
(6) when solution E temperature drops to 40 DEG C, solution D is slowly poured into, mixing.
The store method of hilsa herring sperm above-mentioned, the preparation of dilution the following steps are included:
(1) 100ml pure water is measured, in 105 DEG C of high pressure sterilization 11min, obtains sterilizing pure water A；
(2) lemon sodium bicarbonate, glucose, sodium citrate, Bifidobacterium, streptomysin and lactic acid bacteria are weighed, is added separately to go out
In bacterium pure water A, in 60 DEG C of heating water baths, and stirring and dissolving 12min clockwise is carried out, then in 120 DEG C of high pressure sterilizations
18min obtains solution B；
(3) the sterile fresh yolk liquid of absorption is added in the centrifuge tube of sterilizing after carrying out 1:1 dilution with sterile saline,
It is centrifuged 12min in 5 DEG C of 7500g, draws sodium chloride injection 13ml, in 20 DEG C of standing 10min, yolk is obtained and is centrifuged supernatant
Liquid C；
(4) potassium chloride and calcium chloride are weighed, is added separately in yolk centrifuged supernatant C, stirs and evenly mixs clockwise, is obtained
Solution D；
(5) it takes out solution B and weighs chlorogenic acid, glycerol, lecithin and polyglutamic acid when its temperature drops to 85 DEG C or less
Sodium is added in solution B, and carries out stirring and dissolving clockwise, obtains solution E；
(6) when solution E temperature drops to 38 DEG C, solution D is slowly poured into, mixing.
It is using technical solution of the present invention beneficial effect: maintains hilsa herring sperm activity using active bacteria for the first time, effectively press down
Harmful microbe growth and breeding has been made, has built advantageous active environment for sperm, sperm is delayed to be started to live after oxygen activated
The dynamic time effectively has adjusted liquid environment osmotic pressure, reduces the metabolic rate of sperm, reduces energy consumption, to extend
Sperm life；K+ and Ca2+ is the important adjusting factor of fish sperm vigor, and sperm motility can be improved, and extends the service life；The guarantor
Liquid storage science dilution seminal concentration is moderate, easy to operate, low in cost, can effectively save hilsa herring sperm, can be in production
It is widely used to promote.
Specific embodiment
Embodiment 1
The present embodiment provides a kind of store method of hilsa herring sperm, by antifreeze, hilsa herring sperm and dilution 1:1 by volume:
After 5 mixing, and saved at 5 DEG C；
Dilution volume percent composition are as follows: sodium chloride injection: 50%, pure water: 15%, potassium chloride: 7%, calcium chloride: 5%, carbonic acid
Hydrogen sodium: 5%, chlorogenic acid: 1%, glycerol: 1%, glucose: 3%, lecithin: 8%, sodium citrate: 1%, streptomysin: 1%, polyglutamic acid
Sodium: 1%, Bifidobacterium: 1%, surplus is lactic acid bacteria；
Antifreeze percent by volume ingredient are as follows: dimethyl sulfoxide: 30%, glycerol: 20%, ethylene glycol: 30%, dimethyl acetamide:
20%；
The store method of above-mentioned hilsa herring sperm, the preparation of dilution the following steps are included:
(1) 100ml pure water is measured, in 100 DEG C of high pressure sterilization 10min, obtains sterilizing pure water A；
(2) lemon sodium bicarbonate, glucose, sodium citrate, Bifidobacterium, streptomysin and lactic acid bacteria are weighed, is added separately to go out
In bacterium pure water A, in 55 DEG C of heating water baths, and stirring and dissolving 10min clockwise is carried out, then in 120 DEG C of high pressure sterilizations
15min obtains solution B；
(3) the sterile fresh yolk liquid of absorption is added in the centrifuge tube of sterilizing after carrying out 1:1 dilution with sterile saline,
It is centrifuged 10min in 5 DEG C of 7500g, draws sodium chloride injection 10ml, in 15 DEG C of standing 8min, obtains yolk centrifuged supernatant
C；
(4) potassium chloride and calcium chloride are weighed, is added separately in yolk centrifuged supernatant C, stirs and evenly mixs clockwise, is obtained
Solution D；
(5) it takes out solution B and weighs chlorogenic acid, glycerol, lecithin and polyglutamic acid when its temperature drops to 85 DEG C or less
Sodium is added in solution B, and carries out stirring and dissolving clockwise, obtains solution E；
(6) when solution E temperature drops to 35 DEG C, solution D is slowly poured into, mixing.
Embodiment 2
The present embodiment provides a kind of store method of hilsa herring sperm, by antifreeze, hilsa herring sperm and dilution 1:1 by volume:
After 5 mixing, and saved at 5 DEG C；
Dilution volume percent composition are as follows: sodium chloride injection: 50%, pure water: 20%, potassium chloride: 14%, calcium chloride: 10%, carbon
Sour hydrogen sodium: 10%, chlorogenic acid: 2%, glycerol: 3%, glucose: 5%, lecithin: 15%, sodium citrate: 3%, streptomysin: 2%, poly- paddy
Propylhomoserin sodium: 3%, Bifidobacterium: 3%, surplus is lactic acid bacteria；
Antifreeze percent by volume ingredient are as follows: dimethyl sulfoxide: 30%, glycerol: 20%, ethylene glycol: 30%, dimethyl acetamide:
20%；
The store method of above-mentioned hilsa herring sperm, the preparation of dilution the following steps are included:
(1) 100ml pure water is measured, in 110 DEG C of high pressure sterilization 12min, obtains sterilizing pure water A；
(2) lemon sodium bicarbonate, glucose, sodium citrate, Bifidobacterium, streptomysin and lactic acid bacteria are weighed, is added separately to go out
In bacterium pure water A, in 65 DEG C of heating water baths, and stirring and dissolving 15min clockwise is carried out, then in 120 DEG C of high pressure sterilizations
20min obtains solution B；
(3) the sterile fresh yolk liquid of absorption is added in the centrifuge tube of sterilizing after carrying out 1:1 dilution with sterile saline,
It is centrifuged 15min in 5 DEG C of 7500g, draws sodium chloride injection 15ml, in 25 DEG C of standing 12min, yolk is obtained and is centrifuged supernatant
Liquid C；
(4) potassium chloride and calcium chloride are weighed, is added separately in yolk centrifuged supernatant C, stirs and evenly mixs clockwise, is obtained
Solution D；
(5) it takes out solution B and weighs chlorogenic acid, glycerol, lecithin and polyglutamic acid when its temperature drops to 85 DEG C or less
Sodium is added in solution B, and carries out stirring and dissolving clockwise, obtains solution E；
(6) when solution E temperature drops to 40 DEG C, solution D is slowly poured into, mixing.
Embodiment 3
The present embodiment provides a kind of store method of hilsa herring sperm, by antifreeze, hilsa herring sperm and dilution 1:1 by volume:
After 5 mixing, and saved at 5 DEG C；
Dilution volume percent composition are as follows: sodium chloride injection: 50%, pure water: 18%, potassium chloride: 12%, calcium chloride: 8%, carbon
Sour hydrogen sodium: 8%, chlorogenic acid: 1%, glycerol: 1%, glucose: 3%, lecithin: 10%, sodium citrate: 2%, streptomysin: 1%, polyglutamic
Sour sodium: 2%, Bifidobacterium: 2%, surplus is lactic acid bacteria；
Antifreeze percent by volume ingredient are as follows: dimethyl sulfoxide: 30%, glycerol: 20%, ethylene glycol: 30%, dimethyl acetamide:
20%；
The store method of above-mentioned hilsa herring sperm, the preparation of dilution the following steps are included:
(1) 100ml pure water is measured, in 105 DEG C of high pressure sterilization 11min, obtains sterilizing pure water A；
(2) lemon sodium bicarbonate, glucose, sodium citrate, Bifidobacterium, streptomysin and lactic acid bacteria are weighed, is added separately to go out
In bacterium pure water A, in 60 DEG C of heating water baths, and stirring and dissolving 12min clockwise is carried out, then in 120 DEG C of high pressure sterilizations
18min obtains solution B；
(3) the sterile fresh yolk liquid of absorption is added in the centrifuge tube of sterilizing after carrying out 1:1 dilution with sterile saline,
It is centrifuged 12min in 5 DEG C of 7500g, draws sodium chloride injection 13ml, in 20 DEG C of standing 10min, yolk is obtained and is centrifuged supernatant
Liquid C；
(4) potassium chloride and calcium chloride are weighed, is added separately in yolk centrifuged supernatant C, stirs and evenly mixs clockwise, is obtained
Solution D；
(5) it takes out solution B and weighs chlorogenic acid, glycerol, lecithin and polyglutamic acid when its temperature drops to 85 DEG C or less
Sodium is added in solution B, and carries out stirring and dissolving clockwise, obtains solution E；
(6) when solution E temperature drops to 38 DEG C, solution D is slowly poured into, mixing.
In addition to the implementation, the present invention can also have other embodiments.It is all to use equivalent substitution or equivalent transformation shape
At technical solution, fall within the scope of protection required by the present invention.
Claims (8)
1. a kind of store method of hilsa herring sperm, it is characterised in that: by antifreeze, hilsa herring sperm and dilution by volume 1:
After 1:5 mixing, and saved at 5 DEG C；
The dilution volume percent composition are as follows: sodium chloride injection: 50%, pure water: 15-20%, potassium chloride: 7-14%, chlorination
Calcium: 5-10%, sodium bicarbonate: 5-10%, chlorogenic acid: 1-2%, glycerol: 1-3%, glucose: 3-5%, lecithin: 8-15%, citric acid
Sodium: 1-3%, streptomysin: 1-2%, polyglutamic acid sodium: 1-3%, Bifidobacterium: 1-3%, surplus are lactic acid bacteria；
The antifreeze percent by volume ingredient are as follows: dimethyl sulfoxide: 30%, glycerol: 20%, ethylene glycol: 30%, dimethyl second
Amide: 20%.
2. the store method of hilsa herring sperm according to claim 1, it is characterised in that: the dilution percent by volume at
It is divided into: sodium chloride injection: 50%, pure water: 15%, potassium chloride: 7%, calcium chloride: 5%, sodium bicarbonate: 5%, chlorogenic acid: 1%, it is sweet
Oil: 1%, glucose: 3%, lecithin: 8%, sodium citrate: 1%, streptomysin: 1%, polyglutamic acid sodium: 1%, Bifidobacterium: 1%, it is remaining
Amount is lactic acid bacteria.
3. the store method of hilsa herring sperm according to claim 1, it is characterised in that: the dilution percent by volume at
It is divided into: sodium chloride injection: 50%, pure water: 20%, potassium chloride: 14%, calcium chloride: 10%, sodium bicarbonate: 10%, chlorogenic acid: 2%,
Glycerol: 3%, glucose: 5%, lecithin: 15%, sodium citrate: 3%, streptomysin: 2%, polyglutamic acid sodium: 3%, Bifidobacterium: 3%,
Surplus is lactic acid bacteria.
4. the store method of hilsa herring sperm according to claim 1, it is characterised in that: the dilution percent by volume at
It is divided into: sodium chloride injection: 50%, pure water: 18%, potassium chloride: 12%, calcium chloride: 8%, sodium bicarbonate: 8%, chlorogenic acid: 1%, it is sweet
Oil: 1%, glucose: 3%, lecithin: 10%, sodium citrate: 2%, streptomysin: 1%, polyglutamic acid sodium: 2%, Bifidobacterium: 2%, it is remaining
Amount is lactic acid bacteria.
5. the store method of the hilsa herring sperm as described in any claim in claim 1-4, which is characterized in that the dilution
The preparation of liquid the following steps are included:
(1) 100ml pure water is measured, in 100-110 DEG C of high pressure sterilization 10-12min, obtains sterilizing pure water A；
(2) lemon sodium bicarbonate, glucose, sodium citrate, Bifidobacterium, streptomysin and lactic acid bacteria are weighed, is added separately to go out
In bacterium pure water A, in 55-65 DEG C of heating water bath, and stirring and dissolving 10-15min clockwise is carried out, then gone out in 120 DEG C of high pressures
Bacterium 15-20min obtains solution B；
(3) the sterile fresh yolk liquid of absorption is added in the centrifuge tube of sterilizing after carrying out 1:1 dilution with sterile saline,
It is centrifuged 10-15min in 5 DEG C of 7500g, draws sodium chloride injection 10-15ml, in 15-25 DEG C of standing 8-12min, obtains ovum
Yellow centrifuged supernatant C；
(4) potassium chloride and calcium chloride are weighed, is added separately in yolk centrifuged supernatant C, stirs and evenly mixs clockwise, is obtained
Solution D；
(5) it takes out solution B and weighs chlorogenic acid, glycerol, lecithin and polyglutamic acid when its temperature drops to 85 DEG C or less
Sodium is added in solution B, and carries out stirring and dissolving clockwise, obtains solution E；
(6) when solution E temperature drops to 35-40 DEG C, solution D is slowly poured into, mixing.
6. the store method of hilsa herring sperm according to claim 5, it is characterised in that: the preparation of the dilution include with
Lower step:
(1) 100ml pure water is measured, in 100 DEG C of high pressure sterilization 10min, obtains sterilizing pure water A；
(2) lemon sodium bicarbonate, glucose, sodium citrate, Bifidobacterium, streptomysin and lactic acid bacteria are weighed, is added separately to go out
In bacterium pure water A, in 55 DEG C of heating water baths, and stirring and dissolving 10min clockwise is carried out, then in 120 DEG C of high pressure sterilizations
15min obtains solution B；
(3) the sterile fresh yolk liquid of absorption is added in the centrifuge tube of sterilizing after carrying out 1:1 dilution with sterile saline,
It is centrifuged 10min in 5 DEG C of 7500g, draws sodium chloride injection 10ml, in 15 DEG C of standing 8min, obtains yolk centrifuged supernatant
C；
(4) potassium chloride and calcium chloride are weighed, is added separately in yolk centrifuged supernatant C, stirs and evenly mixs clockwise, is obtained
Solution D；
(5) it takes out solution B and weighs chlorogenic acid, glycerol, lecithin and polyglutamic acid when its temperature drops to 85 DEG C or less
Sodium is added in solution B, and carries out stirring and dissolving clockwise, obtains solution E；
(6) when solution E temperature drops to 35 DEG C, solution D is slowly poured into, mixing.
7. the store method of hilsa herring sperm according to claim 5, it is characterised in that: the preparation of the dilution include with
Lower step:
(1) 100ml pure water is measured, in 110 DEG C of high pressure sterilization 12min, obtains sterilizing pure water A；
(2) lemon sodium bicarbonate, glucose, sodium citrate, Bifidobacterium, streptomysin and lactic acid bacteria are weighed, is added separately to go out
In bacterium pure water A, in 65 DEG C of heating water baths, and stirring and dissolving 15min clockwise is carried out, then in 120 DEG C of high pressure sterilizations
20min obtains solution B；
(3) the sterile fresh yolk liquid of absorption is added in the centrifuge tube of sterilizing after carrying out 1:1 dilution with sterile saline,
It is centrifuged 15min in 5 DEG C of 7500g, draws sodium chloride injection 15ml, in 25 DEG C of standing 12min, yolk is obtained and is centrifuged supernatant
Liquid C；
(4) potassium chloride and calcium chloride are weighed, is added separately in yolk centrifuged supernatant C, stirs and evenly mixs clockwise, is obtained
Solution D；
(5) it takes out solution B and weighs chlorogenic acid, glycerol, lecithin and polyglutamic acid when its temperature drops to 85 DEG C or less
Sodium is added in solution B, and carries out stirring and dissolving clockwise, obtains solution E；
(6) when solution E temperature drops to 40 DEG C, solution D is slowly poured into, mixing.
8. the store method of hilsa herring sperm according to claim 5, it is characterised in that: the preparation of the dilution include with
Lower step:
(1) 100ml pure water is measured, in 105 DEG C of high pressure sterilization 11min, obtains sterilizing pure water A；
(2) lemon sodium bicarbonate, glucose, sodium citrate, Bifidobacterium, streptomysin and lactic acid bacteria are weighed, is added separately to go out
In bacterium pure water A, in 60 DEG C of heating water baths, and stirring and dissolving 12min clockwise is carried out, then in 120 DEG C of high pressure sterilizations
18min obtains solution B；
(3) the sterile fresh yolk liquid of absorption is added in the centrifuge tube of sterilizing after carrying out 1:1 dilution with sterile saline,
It is centrifuged 12min in 5 DEG C of 7500g, draws sodium chloride injection 13ml, in 20 DEG C of standing 10min, yolk is obtained and is centrifuged supernatant
Liquid C；
(4) potassium chloride and calcium chloride are weighed, is added separately in yolk centrifuged supernatant C, stirs and evenly mixs clockwise, is obtained
Solution D；
(5) it takes out solution B and weighs chlorogenic acid, glycerol, lecithin and polyglutamic acid when its temperature drops to 85 DEG C or less
Sodium is added in solution B, and carries out stirring and dissolving clockwise, obtains solution E；
(6) when solution E temperature drops to 38 DEG C, solution D is slowly poured into, mixing.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110419471A (en) * | 2019-09-05 | 2019-11-08 | 中国水产科学研究院淡水渔业研究中心 | A kind of method of wild Burma hilsa herring artificial insemination and incubating oosperm |
| CN114375946A (en) * | 2022-01-22 | 2022-04-22 | 上海市农业科学院 | Cryopreservation method of chicken semen |
| CN114651815A (en) * | 2022-04-12 | 2022-06-24 | 江苏农牧科技职业学院 | Method for preserving frozen boar semen |
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| CN114375946A (en) * | 2022-01-22 | 2022-04-22 | 上海市农业科学院 | Cryopreservation method of chicken semen |
| CN114651815A (en) * | 2022-04-12 | 2022-06-24 | 江苏农牧科技职业学院 | Method for preserving frozen boar semen |
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