CN106259089A - A kind of method of large-scale production gynoecial loaches - Google Patents
A kind of method of large-scale production gynoecial loaches Download PDFInfo
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Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K61/00—Culture of aquatic animals
-
- 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
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- Life Sciences & Earth Sciences (AREA)
- Environmental Sciences (AREA)
- Marine Sciences & Fisheries (AREA)
- Zoology (AREA)
- Animal Husbandry (AREA)
- Biodiversity & Conservation Biology (AREA)
- Farming Of Fish And Shellfish (AREA)
Abstract
A kind of method that the invention discloses large-scale production gynoecial loaches; step includes: large-scale production Misgurni anguillicaudati; the healthy excellent raun of screening is as standby parent; and carry out antenatal reinforced cultivating and artificial induced spawning; Heterologous Sperm is used to stimulate and the gynogenesis method of cold shock treatment, it is thus achieved that complete female diploid seed；The artemia larvae crossed by Methyl testosterone 50mg/kg androgen solution soaking makees bait, continues 30 days, and in mud fish fry, XX ♀ raun sexual inversion becomes XX ♂ physiology milter；The complete normal fingerling of female diploid and above-mentioned physiology milter are carried out copulation, large-scale production 100% gynoecial loaches.Compared with prior art, gynogenesis and sexual inversion technology are combined by the present invention, and large-scale production gynoecial loaches increases substantially per mu yield and culture benefit.Gynogenesis technology can also comparatively fast set up pure lines, provides very abundant material for genetic breeding, shortens the selection-breeding cycle of gynoecial loaches, it is adaptable to large-scale Production requirement, it is easier to popularization and application.
Description
Technical field
The present invention relates to the technical field of a kind of aquaculture, in particular a kind of complete female mud of large-scale production
The method of loach.
Background technology
Misgurni anguillicaudati (Misgurnus arguillicaudatus) is the treasure of Aquatic product, and market demand breach is big, and price is every year
Soaring.Misgurni anguillicaudati fine and tender taste, delicious flavour, nutritious, and rich in mineral substances and multivitamin, there is multiple medicinal valency
Value, wherein vitamin B content is higher 3～4 times than Channa argus, shrimps, among the people have saying of " Radix Ginseng in water ".Loach Protein matter content is equal
Higher than most conventional cultured fishes, amino acid content composition is more considered the good merchantable brand of nourishing and fit keeping function comprehensively, is processing animals egg
One of best raw material of white lead.
Misgurni anguillicaudati is the important Fish of freshwater aquiculture, and in nature, Misgurni anguillicaudati raun is bigger compared to milter individuality.Identical
Cultivating condition under, female Misgurni anguillicaudati is faster than the growth of milter born of the same parents, quality better, more popular with consumers.Cultivation gynoecial loaches will
Significantly improve cultured output and economic benefit.Produce gynoecial loaches and there is several methods that available: such as artificial selection, estrogen
Directly inductivity reverses and gynogenesis etc..Manual selection techniques is big because of labor intensity, and can not accurate district during seed
Divide female milter, be not suitable for the large-scale production of unisexuality raun.Estrogen direct inductivity reversal method can make hero with scale
Property piscinity reverse and become female Misgurni anguillicaudati, but this method can not reach the most female purpose, and the danger of sexual hormone residues, is unfavorable for
Safety in production, already by many countries disabling.
Chinese patent literature (CN103548740A) discloses a kind of method producing gynoecial loaches, and the method is by mud
Loach carries out the technology such as artificial sexual inversion, testcross and screening and group expanding to control the female loach breeding of unisexuality, to provide the one can scale
The gynoecial loaches cultural method produced, improves Misgurni anguillicaudati yield and commercial quality.But, the method screening cycle is long, screening side
Method is relatively complicated, the highest for its human cost of large-scale farming and time cost.
It is known that Sex Determination Mechanism of Fish is sufficiently complex.Owing to heterosomal differentiation is also in original condition, very
There is heteromorphism chromosome in few kind, so identifying that from cytology heterosomal existence is almost unlikely.In day
This, the Sex Determination Mechanism of Misgurni anguillicaudati is the most studied, is the most tentatively judged as female same distribution type i.e. XX/XY type.Artificial induction is female
Caryogenesis is a very important fishery breeding technology, and at home, gynogenesis technology starts from 1981, Wu Qingjiang et al.
Utilize cold shock to process 2 kinds of different methods with Colchicine and obtain Cyprinus carpio gynogenesis offspring.
Summary of the invention
It is an object of the invention to overcome the deficiencies in the prior art, it is provided that the side of a kind of large-scale production gynoecial loaches
Method, to provide the breeding method of a kind of simpler, safe gynoecial loaches.
The present invention is achieved by the following technical solutions:
A kind of method that the invention provides large-scale production gynoecial loaches, comprises the following steps:
(1) parent culture: large-scale cultivation Misgurni anguillicaudati, the female loach breeding of screening build grace fast growth is as standby parent
This；
(2) gynogenesis: female loach parent uses red Carassius auratus Heterologous Sperm stimulate and cold shock treatment method carries out thelykaryon
Growth processes, it is thus achieved that complete female diploid seed；
(3) sexual inversion: the complete female diploid seed of part just hatching is reversed with androgen Progressive symmetric erythrokeratodermia, it is thus achieved that physiology is male
Fish；
(4) large-scale production: with the normal fingerling of complete female diploid of step (2) for female parent, with the physiology milter of step (3)
Copulation, large-scale production 100% gynoecial loaches is carried out for male parent.
Further, described gynogenetic method is:
(I) paradise fish of China sperm utilizing inactivation stimulates the ovum of female loach, it is thus achieved that " germ cell "；
(II) cold shock treatment 25min at " germ cell " of step (I) being placed in 0-4 DEG C, then cultivated by conventional method
Obtain the complete normal fingerling of female diploid.
Further, in described step (I), the method irradiated with ultraviolet carries out inactivation treatment, and described inactivation treatment can
To use following method: the red Carassius auratus seminal fluid of dilution being taken 4mL and is laid on minute surface, be 15W × 2 in intensity, range is every time
Irradiate 60s, test under microscope motility of sperm under the ultraviolet of 28-30cm, determine inactivation situation.But the intensity that ultraviolet irradiates
It is not limited to this with range and irradiation time, experiment can adjust according to motility of sperm ultraviolet irradiation condition in real time.
Further, the method for described sexual inversion is: with artemia larvae for androgen carrier, is soaked by androgen solution
The artemia larvae steeped is thrown something and fed as the starter diet of fry, after throwing something and feeding 30-60 days continuously, transfers normal raising to；Through hero
In the fry that gonadal hormone is survived after processing, part raun sexual inversion becomes physiology milter, but also has the female of the non-sexual inversion of part
Fish, due to raun pectoral fin short and justify, milter pectoral fin length and sharp, both profile difference are obvious, therefore, when fry is cultivated to pressing
When pectoral fin shape distinguishes male and female, screening obtains physiology milter and cultivates by normal method for breeding, as the male parent producing gynoecial loaches.
Further, described androgen is Methyl testosterone.
Further, in described step (4), before copulation, respectively to the complete female Misgurni anguillicaudati of female diploid and physiological male Misgurni anguillicaudati
Oxytocin injection, then according to put in a suitable place to breed maternal mixing with the ratio of male parent mantissa 1:2, allows its spawning and fertilization voluntarily.
Further, male parent and female parent being placed in net cage of laying eggs and carry out mixing and put in a suitable place to breed, breeding density is 2.5kg/m2。
Further, described in lay eggs net cage be additionally provided with fish nest, make fish roe after fertilization can stick on fish nest；Lay eggs net
Setting circulating water outside case, take batch (-type) stimulation by running water to promote Misgurni anguillicaudati to lay eggs voluntarily in net cage of laying eggs, batch (-type) stimulation by running water is just
Lay eggs voluntarily in Misgurni anguillicaudati.
Further, treat that male parent and female parent after natural spawning, first take out fish nest, then pull puerperal out in net cage
Misgurni anguillicaudati, then wash the germ cell of lower attachment, by germ cell after container metering ovum number, put in hatching cylinder and hatch, incubate to improve
Rate.
The present invention has the advantage that the side that the invention provides a kind of large-scale production gynoecial loaches compared to existing technology
Method, gynogenesis and sexual inversion technology are combined by the method, in order to research and develop gynoecial loaches, can increase substantially loach breeding list
Producing and quality, gynogenesis technology can comparatively fast set up pure lines, can provide, for genetic breeding, the material enriched very much, be greatly shortened
The cultivation cycle of gynoecial loaches, simplifies cultural method, has certain optimization, especially on time cost and human cost
It is applicable to large-scale Production requirement, it is easier to popularization and application.
Detailed description of the invention
Elaborating embodiments of the invention below, the present embodiment is carried out under premised on technical solution of the present invention
Implement, give detailed embodiment and concrete operating process, but protection scope of the present invention is not limited to following enforcement
Example.
Embodiment 1
A kind of method present embodiments providing large-scale production gynoecial loaches, comprises the following steps:
(1) parent culture
Large-scale production loach fry, the healthy raun of screening as standby parent, particularly as follows:
1. cultivating 7-10d, fry total length reaches 1.5-2.0cm, screens once, big specification fry of selecting and remain, rate 50% of selecting and remain；
2. will stay and select fry transfer to put in a suitable place to breed, breeding density is 4000 tails/m2, continuing to cultivate 7-10d, fry total length reaches 2-
3cm, screens once, big specification fry of selecting and remain, rate 50% of selecting and remain；
3. adjusting breeding density is 2000 tails/m2, continuing to cultivate 7-10d, fry total length reaches 3-5cm, and screening once, is selected and remain
Big specification fry, the rate of selecting and remain is 50%, it is thus achieved that standby parent；
4. winter sheet selects: after winter cleans up the pond, and screening specification is big, type standard, healthy individuals are reserved seed for planting, and stays 20% as standby
Parent culture.
(2) gynogenesis
(I) open Burdick lamp in advance to irradiate, make Wavelength stabilized.Take the red of 4 times of 0.65% (quality) normal saline dilution
Cyprinus carpio seminal fluid is laid on minute surface, is placed under ultraviolet irradiation 60s, test under microscope motility of sperm, determines inactivation situation.With
Time take the ovum of raun parent, carry out artificial fertilization with the sperm of inactivation, it is thus achieved that " germ cell "." germ cell " is different from often herein
Rule germ cell, for pseudogamy ovum, should only comprise maternal chromosome in " germ cell ".
(II) cold shock treatment 25min at " germ cell " of step (I) being placed in 0-4 DEG C, induction obtains complete female diploid,
Complete female diploid is supported to sexual maturity by conventional culture methods, and the Misgurni anguillicaudati as follow-up large-scale production is maternal.
(3) sexual inversion
By above-mentioned gynogenetic complete female diploid loach fry 50mg/kg androgen solution (Methyl testosterone
Artemia larvae 50mg/kg) soaked is thrown something and fed continuously as starter diet, after continuing 30 days, and part in the fry of survival
Raun sexual inversion becomes physiology milter, after artificial breeding is 10 months, according to pectoral fin feature, selects male Misgurni anguillicaudati as follow-up rule
Modelling produces the male parent of gynoecial loaches.
(4) large-scale production
Before (i) copulation, female parent and the male parent oxytocin injection of step (3) to step (2), the formula bag of described oxytocin
Including: the DOM of LHRH-A2 and 1mg/kg of HCG, 2ug/kg of 125IU/kg, maternal every endnote penetrates 0.1-0.2mL, the every tail of male parent
Injection 0.05-0.1mL.
(ii) by having injected the female parent of oxytocin and male parent according to the ratio combo that mantissa is 1:2, it is placed in net cage and carries out
Mixing is put in a suitable place to breed, and breeding density controls at 2.5kg/m2, the bottom of each net cage is placed multiple fish nests, is had circulating water, net outside net cage
Rushing water filling with little leather hose in case, take batch (-type) stimulation by running water to promote Misgurni anguillicaudati to lay eggs voluntarily, now female parent and male parent are in net cage
Spawning and fertilization voluntarily, it is thus achieved that germ cell be attached on fish nest, utilize net cage and fish nest to cultivate Misgurni anguillicaudati, create suitably to Misgurni anguillicaudati
Breeding environment, enable Misgurni anguillicaudati to lay eggs voluntarily, simplify Misgurni anguillicaudati propagation method, decrease labour cost.Through statistics, utilize this
Application method cultivate gynoecial loaches time, its spawning rate more than 80%, rate of fertilization more than 85%.
(iii), after treating natural spawning, take out fish nest, wash the germ cell of lower attachment, after container metering ovum number, will
Germ cell is put in hatching cylinder and is carried out Lotic hatching, final acquisition 100% gynoecial loaches.
So far, lasting large-scale production process and their the breeding system of gynoecial loaches are i.e. defined.This brood body
The foundation of system only must time of 2 years, after obtaining complete female diploid seed the most then, Second Year can be accomplished scale production, greatly
Shorten greatly the cultivation cycle of gynoecial loaches.After Establishing, the per mu yield of gynoecial loaches improves than compareing common Misgurni anguillicaudati
More than 30%, cultivation per mu yield and culture benefit are greatly improved.
Above example is merely to the preferably embodiment that absolutely proves the present invention and enumerate, the protection model of the present invention
Enclosing and be not limited to this, those skilled in the art's equivalent done by the basis of the present invention substitutes or conversion, all the present invention's
Within protection domain.
Claims (10)
1. the method for a large-scale production gynoecial loaches, it is characterised in that comprise the following steps:
(1) parent culture: large-scale cultivation Misgurni anguillicaudati, the female loach breeding of screening build grace fast growth is as standby parent；
(2) gynogenesis: female loach parent uses red Carassius auratus Heterologous Sperm stimulate and cold shock treatment method carries out gynogenesis
Process, it is thus achieved that complete female diploid seed；
(3) sexual inversion: the complete female diploid seed of part just hatching is reversed with androgen Progressive symmetric erythrokeratodermia, it is thus achieved that physiology milter；
(4) large-scale production: with the normal fingerling of complete female diploid of step (2) for maternal, with the physiology milter of step (3) as father
Originally copulation, large-scale production 100% gynoecial loaches are carried out.
The method of a kind of large-scale production gynoecial loaches the most according to claim 1, it is characterised in that described gynogenesis
Method be:
(I) paradise fish of China sperm utilizing inactivation stimulates the ovum of female loach, it is thus achieved that " germ cell "；
(II) cold shock treatment 25min at " germ cell " of step (I) being placed in 0-4 DEG C, then cultivate acquisition by conventional method
The complete normal fingerling of female diploid.
The method of a kind of large-scale production gynoecial loaches the most according to claim 2, it is characterised in that described step (I)
In, the method irradiated with ultraviolet carries out inactivation treatment to red Carassius auratus sperm.
The method of a kind of large-scale production gynoecial loaches the most according to claim 3, it is characterised in that described inactivation treatment
Method be: the red Carassius auratus seminal fluid of dilution being taken 4mL every time and is laid on minute surface, is 15W × 2 in intensity, range is 28-30cm
Ultraviolet under irradiate 60s, test under microscope motility of sperm, determine inactivation situation.
The method of a kind of large-scale production gynoecial loaches the most according to claim 1, it is characterised in that described sexual inversion
Method is: with artemia larvae for androgen carrier, and artemia larvae androgen solution soaking crossed is as the opening of fry
Feedstuff is thrown something and fed, and after throwing something and feeding 30-60 days continuously, transfers normal raising to；Wait raising to when can press pectoral fin shape differentiation male and female, choose
Select physiology milter as standby male parent, and carry out reinforced cultivating.
The method of a kind of large-scale production gynoecial loaches the most according to claim 5, it is characterised in that described androgen
For Methyl testosterone.
The method of a kind of large-scale production gynoecial loaches the most according to claim 1, it is characterised in that described step (4)
In, before copulation, respectively to the complete female Misgurni anguillicaudati of female diploid and physiological male loach injection oxytocin, then according to maternal and male parent
The ratio mixing of mantissa 1:2 is put in a suitable place to breed, allows its spawning and fertilization voluntarily.
The method of a kind of large-scale production gynoecial loaches the most according to claim 1, it is characterised in that described step (4)
In, male parent and female parent being placed in net cage of laying eggs and carries out mixing and put in a suitable place to breed, breeding density is 2.5kg/m2。
The method of a kind of large-scale production gynoecial loaches the most according to claim 8, it is characterised in that described in lay eggs net cage
In be additionally provided with fish nest, set circulating water outside net cage of laying eggs, in net cage of laying eggs, take batch (-type) stimulation by running water to promote Misgurni anguillicaudati to produce voluntarily
Ovum.
The method of a kind of large-scale production gynoecial loaches the most according to claim 9, it is characterised in that treat male parent and mother
This is in net cage after natural spawning, first takes out fish nest, then pulls the Misgurni anguillicaudati in puerperal out, then washes the germ cell of lower attachment, will
Germ cell, after container metering ovum number, is put in hatching cylinder and is hatched.
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| CN107232092A (en) * | 2017-05-26 | 2017-10-10 | 武汉百瑞生物技术有限公司 | A kind of production method of loach pseudo-milter |
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| CN109769722A (en) * | 2019-03-25 | 2019-05-21 | 华中农业大学 | A method of gynoecial loaches being quickly mass produced without using sex hormone |
| CN112262794A (en) * | 2020-10-24 | 2021-01-26 | 武汉市农业科学院 | Method for sustainable large-scale production of all-female mandarin fish |
| CN115428752A (en) * | 2022-10-12 | 2022-12-06 | 华中农业大学 | Method for producing all-female grass carp in large scale and application |
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