CN101869512A - Method for collecting semen of primates of macaque by electro stimulation - Google Patents
Method for collecting semen of primates of macaque by electro stimulation Download PDFInfo
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- CN101869512A CN101869512A CN 201010198699 CN201010198699A CN101869512A CN 101869512 A CN101869512 A CN 101869512A CN 201010198699 CN201010198699 CN 201010198699 CN 201010198699 A CN201010198699 A CN 201010198699A CN 101869512 A CN101869512 A CN 101869512A
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Abstract
The invention provides a method for collecting semen of primates of macaque by electro stimulation, belonging to the method of animal reproduction, in particular to the method for collecting sperm of primates of macaque. In the invention, aluminium foil electrodes coated by absorbent cotton infiltrated by saline encircles around the penis of the anesthetized animal and is connected with the impulse power; the voltage begins from 0V, and stimulation is conducted for 10-15s continuously when each 10V is increased, until the voltage reaches 60V; the process is circulated for 6 to 7 times when necessary, with an interval of 20-30s between two stimulations; and the collected semen can be used for fertilization after incubation and separation. The invention can improve the success rate for collecting semen of animals, and has no pollution; the semen collected in the invention has similar index as that of the semen gained through natural mating and can be applied to external fertilization and artificial fertilization.
Description
Technical field
The invention belongs to animal reproduction method, the method for particularly gathering the macaca primate sperm.
Background technology
Macaca primate, as macaque, machin, Assamese macaque, macaca thibetana etc. is human close relative, its organizational structure, aspects such as Physiology and biochemistry, immunity and metabolism are highly similar to the mankind, are the best laboratory animals of setting up researchs such as infectious disease model, spiritual nervous system disease model, cardiovascular disease model, tumor model and reproductive physiology such as virus.But because the lasting deterioration and the destruction of field survivorship environment, wild primate fertility and essential condition descend gradually and lose, and each primate species has become rare species in the Macaca, and is listed in Class 1 or 2 species under State protection.Along with the increase of population and the deterioration of environment, the human on the one hand continuous puzzlement that faces some great functional difficult diseases, need a large amount of macaque animals to be used for physicism and solution population and health problem, on the other hand, raising of these animals and breeding and corresponding facility etc. are but very expensive.Therefore, protection and preservation laboratory animal resource are promoted to the resource requirement of national strategy Journal of Sex Research.Utilization reproduction ancillary technique, external fertilization for example, technology such as artificial insemination can improve utilization rate and the value of male monkey effectively, and can shorten the breeding cycle and improve breeding effectively and produce the efficient of animal, reduce pathophoresis.
The electricity irritation collecting semen is the primate artificial insemination, the basic means of external fertilization and sperm biological study.The present semen collection method commonly used of primate has rectum electro-ejaculation method and penis electro-ejaculation method.The rectal distention stimulation method needs flushing to get rid of animal internal rectum feces before semen collection, electrode accurately need be positioned over prostatic during collection, and this position stimulates organ such as bladder easily and causes seminal fluid by urine pollution, the loaded down with trivial details and unhygienic health of this method step.How conventional primate penis electrostimulation directly twines the penis of animal with aluminium foil, copper mesh etc., metal electrode directly contacts with skin, because such animal penis structure difference, contact condition between penis and the electrode and penis epidermis impedance difference, nearly glans penis end of macaca thibetana and Assamese macaque penis and glans penis trailing edge dorsal part are distributed with intensive thorn-like cutin nipple, make metal electrode and penile surface even contact possibility hardly, be easy to generate local current and cross ambassador's penis epidermis burn even epithelial damage.Therefore, improve the success rate of macaca primate semen collection and to avoid operating process that animal is caused damage be this area problem demanding prompt solution.
Summary of the invention
In order to overcome the prior art complex steps and to pollute drawbacks such as the collection or the animal penis epidermis of easily burning, the purpose of this invention is to provide a kind of penis electricity irritation semen collection method that is widely used in each primate species of Macaca, improve semen collection success rate and acquisition quality.
The object of the invention realizes by following steps:
(1) sexually matured primate is slightly anaesthetized fixation of animal backstroke, cleaning, sterilization penis zone with the intramuscular injection of 3-4mg/kg body weight with ketalar;
(2) get two through the absorbent cotton that normal saline soaks into fully that 2cm is wide, 4-5cm is long, wrap up two 3-5mm aluminium foil paper slips wide, that 6-7cm is long respectively and make two irritation of penis electrode strips;
(3) electrode strip is looped around animal penis base portion and nearly glans penis end respectively, electrode strip is connected with the pulse power with lead around junction point;
(4) make pulse power supply, pulse frequency is set to very low frequency; Voltage, whenever increases progressively and once continues stimulation 10-15s, until 60V with the 5-10Vs speed increase from 0V;
(5) ejaculation promptly stops to stimulate, and collects seminal fluid and hatches 30 minutes in 37 ℃, isolates the seminal fluid of liquid part.
The increasing velocity of described pulse voltage is 10V/s.
Described pulse voltage is a symmetrical square wave, and its frequency is 17Hz, and pulsewidth is 17ms.
Described animal penis base electrode connects pulse power positive pole, and nearly glans penis termination electrode connects pulse power negative pole.
Described method (4) is set by step carried out 6-7 stimulation, interval 20-30s between per twice stimulation.
The progress that the present invention has is:
The present invention utilizes normal saline infiltration absorbent cotton parcel aluminium foil to make with penis good electrodes in contact bar; make that the electric conductivity of electrode is unaffected; and avoided the phenomenon of the excessive penis of burning of local current, have the excellent protection effect for animal penis epidermis.
The present invention has also determined the required voltage range of macaca primate penis electricity irritation semen collection, and basic parameters such as blanking time, pulse frequency, pulsewidth, dutycycle can improve the success rate and the quality of semen collection.
The present invention and environmental friendliness, watch for animals and draw materials easily, easy and simple to handle, save material.
Below by specific embodiment technical scheme of the present invention is described further, embodiment should be understood that annotation of the present invention but not limiting the scope of the invention.
The specific embodiment
Embodiment 1
Slightly anaesthetizing Assamese macaque with ketalar with the intramuscular injection of 4mg/kg body weight, is backstroke with fixation of animal.Clear up monkey penis zone with normal saline, and penile surface is disinfected with 75% medical alcohol.Prepare the aluminium foil paper slips that two 5mm are wide, 7cm is long,, and fully soak into respectively with normal saline and to make two electrode strips respectively with the absorbent cotton parcel.These two electrode strips are looped around the penis base portion and nearly glans penis end of animal respectively, electrode strip is fully contacted with monkey penile surface skin.The positive pole and the negative pole that electrode strip are connected the pulse output current by lead respectively.With the frequency configuration of pulse current is 17Hz, and pulsewidth is 17ms, dutycycle 1, and energized, voltage is from 0V, and with 5V/s speed boosted voltage, every increase 10V continues to stimulate 15s, and then increases 5V until 60V with identical speed.Can carry out 6-7 time if necessary stimulates, the time of having a rest of interval 30s between per twice stimulation.The monkey ejaculation then stops to stimulate immediately.
The Assamese macaque penis epidermis phenomenon of burning does not take place in the Assamese macaque seminal fluid of gathering.The seminal fluid volume is 3.2mL, and wherein liquid partly is 1.2mL, and sperm concentration is 11.5 * 10
8/ mL, sperm motility degree are 81%, and the normal sperm of form accounts for 74%.
Embodiment 2
Slightly anaesthetizing macaque with ketalar with the intramuscular injection of 3mg/kg body weight, is backstroke with fixation of animal.Clear up monkey penis zone with normal saline, and penile surface is disinfected with 75% medical alcohol.Prepare the aluminium foil paper slips that two 3mm are wide, 5cm is long,, and fully soak into respectively with normal saline and to make two electrode strips respectively with the absorbent cotton parcel.These two electrode strips are looped around the penis base portion and nearly glans penis end of animal respectively, electrode strip is fully contacted with monkey penile surface skin.Electrode strip is connected with the pulse current delivery outlet by lead.The penis base electrode connects anodal, and nearly glans penis termination electrode connects negative pole.With the frequency configuration of output square wave is 15Hz, and pulsewidth is 17ms, dutycycle 1, and energized, voltage is from 0V, and with 10V/s speed boosted voltage, every increase 10V continues to stimulate 10s, increases progressively until 60V with 10V again.Repeating 6-7 time stimulates, the time of having a rest of interval 30s between per twice stimulation.Stop immediately stimulating during the monkey ejaculation.
The macaque penis epidermis phenomenon of burning does not take place in the macaque seminal fluid of gathering.The seminal fluid volume is 2mL, and wherein liquid partly is 0.7mL, and sperm concentration is 12.6 * 10
8/ mL, sperm motility degree are 85%, and the normal sperm of form accounts for 78%.
Embodiment 3.
Slightly anaesthetizing machin with ketalar with the intramuscular injection of 3mg/kg body weight, is backstroke with fixation of animal.Clear up monkey penis zone with normal saline, and penile surface is disinfected with 75% medical alcohol.Prepare the aluminium foil paper slips that two 3mm are wide, 5cm is long,, and fully soak into respectively with normal saline and to make two electrode strips respectively with the absorbent cotton parcel.These two electrode strips are looped around the penis base portion and nearly glans penis end of animal respectively, electrode strip is fully contacted with monkey penile surface skin.Electrode strip is connected with the pulse current delivery outlet by lead.The penis base electrode connects anodal, and nearly glans penis termination electrode connects negative pole.The frequency configuration of pulse being exported square wave is 18Hz, and pulsewidth is 17ms, and dutycycle 1, starting voltage are 0V, energized, and with the speed boosted voltage of 10V/s, every increase 10V continues to stimulate 10s, and then increases 10V until 60V with identical speed.Repeating 6-7 time stimulates, the time of having a rest of interval 30s between per twice stimulation.Stop immediately stimulating during the monkey ejaculation.
The machin penis epidermis phenomenon of burning does not take place in the macaque seminal fluid of gathering.The seminal fluid volume is 1.8mL, and wherein liquid partly is 0.5mL, and sperm concentration is 10.5 * 10
8/ mL, sperm motility degree are 79%, and the normal sperm of form accounts for 74%.
Embodiment 4:
Slightly anaesthetizing macaca thibetana with ketalar with the intramuscular injection of 3mg/kg body weight, is backstroke with fixation of animal.Clear up monkey penis zone with normal saline, and penile surface is disinfected with 75% medical alcohol.Prepare the aluminium foil paper slips that two 3mm are wide, 5cm is long,, and fully soak into respectively with normal saline and to make two electrode strips respectively with the absorbent cotton parcel.These two electrode strips are looped around the penis base portion and nearly glans penis end of animal respectively, electrode strip is fully contacted with monkey penile surface skin.Electrode strip is connected pulse current delivery outlet positive pole and negative pole respectively by lead.Pulse output is set to square wave, frequency 17Hz, and pulsewidth is 17ms, dutycycle 1.Energized, voltage are from 0V, and with the speed boosted voltage of 10V/s, every increase 10V continues to stimulate 10s, increases 10V until 60V with identical speed again.Repeating 6-7 time stimulates, the time of having a rest of interval 30s between per twice stimulation.Stop immediately stimulating during the monkey ejaculation.
The macaca thibetana penis epidermis phenomenon of burning does not take place in the macaca thibetana seminal fluid of gathering.The seminal fluid volume is 6.3mL, and wherein liquid partly is 2.1mL, and sperm concentration is 45.6 * 10
8/ mL, sperm motility degree are 82%, and the normal sperm of form accounts for 78%.
Claims (9)
1. method for collecting semen of primates of macaque by electro stimulation comprises the steps:
(1) sexually matured primate is slightly anaesthetized fixation of animal backstroke, cleaning, sterilization penis zone with the intramuscular injection of 3-4mg/kg body weight with ketalar;
(2) get two through the absorbent cotton that normal saline soaks into fully that 2cm is wide, 4-5cm is long, wrap up two 3-5mm aluminium foil paper slips wide, that 6-7cm is long respectively and make two irritation of penis electrode strips;
(3) electrode strip is looped around animal penis base portion and nearly glans penis end respectively, electrode strip is connected with the pulse power with lead around junction point;
(4) make pulse power supply, pulse frequency are very low frequency, and voltage, whenever increases progressively and once continues stimulation 10-15s, until 60V with the 5-10V/s speed increase from 0V;
(5) ejaculation promptly stops to stimulate, and collects seminal fluid and hatches 30 minutes in 37 ℃, isolates the seminal fluid of liquid part.
2. acquisition method according to claim 1, the increasing velocity that it is characterized in that pulse voltage is 10V/s.
3. according to claim 1,2 described acquisition methods, it is characterized in that pulse voltage is a symmetrical square wave, its frequency is 17Hz, and pulsewidth is 17ms.
4. according to claim 1,2 described acquisition methods, it is characterized in that animal penis base electrode connects pulse power positive pole, nearly glans penis termination electrode connects pulse power negative pole.
5. acquisition method according to claim 3 is characterized in that animal penis base electrode connects pulse power positive pole, and nearly glans penis termination electrode connects pulse power negative pole.
6. according to claim 1,2 described acquisition methods, it is characterized in that set by step (4) carry out 6-7 time and stimulate, 20-30s at interval between per twice stimulation.
7. acquisition method according to claim 3 is characterized in that set by step (4) carry out 6-7 time and stimulate, 20-30s at interval between per twice stimulation.
8. acquisition method according to claim 4 is characterized in that set by step (4) carry out 6-7 time and stimulate, 20-30s at interval between per twice stimulation.
9. acquisition method according to claim 5 is characterized in that set by step (4) carry out 6-7 time and stimulate, 20-30s at interval between per twice stimulation.
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| CN 201010198699 CN101869512A (en) | 2010-06-12 | 2010-06-12 | Method for collecting semen of primates of macaque by electro stimulation |
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| CN 201010198699 CN101869512A (en) | 2010-06-12 | 2010-06-12 | Method for collecting semen of primates of macaque by electro stimulation |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103784214A (en) * | 2012-10-30 | 2014-05-14 | 苏州药明康德新药开发有限公司 | Monkey semen collection method |
| CN104758082A (en) * | 2015-03-06 | 2015-07-08 | 华南农业大学 | Semen collection method of macaque |
| CN107669370A (en) * | 2016-08-02 | 2018-02-09 | 西南大学 | The optimization method of one elite stand musk deer electro-ejaculation |
-
2010
- 2010-06-12 CN CN 201010198699 patent/CN101869512A/en active Pending
Non-Patent Citations (1)
| Title |
|---|
| 《动物学研究》 19941231 杨上川等 一种改进的阴茎电刺激采精法和猕猴藏酋猴及熊猴的采精及其精液特征的初步研究 77-83 1-3 第15卷, 第1期 2 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103784214A (en) * | 2012-10-30 | 2014-05-14 | 苏州药明康德新药开发有限公司 | Monkey semen collection method |
| CN103784214B (en) * | 2012-10-30 | 2016-05-25 | 苏州药明康德新药开发股份有限公司 | The method of monkey semen collection |
| CN104758082A (en) * | 2015-03-06 | 2015-07-08 | 华南农业大学 | Semen collection method of macaque |
| CN107669370A (en) * | 2016-08-02 | 2018-02-09 | 西南大学 | The optimization method of one elite stand musk deer electro-ejaculation |
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Application publication date: 20101027 |