CN113229223B

CN113229223B - Reproduction method of centipedes hastellae

Info

Publication number: CN113229223B
Application number: CN202110703311.4A
Authority: CN
Inventors: 岳建军; 杨涛; 严珍
Original assignee: Dai Medical College Of West Yunnan University Of Applied Technology; Institute of Medicinal Plant Development of CAMS and PUMC
Current assignee: Dai Medical College Of West Yunnan University Of Applied Technology; Institute of Medicinal Plant Development of CAMS and PUMC
Priority date: 2021-06-24
Filing date: 2021-06-24
Publication date: 2022-11-22
Anticipated expiration: 2041-06-24
Also published as: CN113229223A

Abstract

The invention belongs to the technical field of medicinal animal feeding, and particularly relates to a centipede breeding method, which is simple to operate, simple and easy to operate in breeding steps, and easy to obtain breeding tools, so that the breeding cost is saved, the breeding period can be effectively shortened, the survival rate and the sub-algebra quantity of centipedes can be obviously improved, and the fatality rate is reduced; meanwhile, the centipede hastelling bred by the method has excellent offspring biological indexes, relatively consistent growth vigor, active and robust individuals and good quality, can meet the primary standard requirements of traditional Chinese medicinal materials, can provide stable and reliable animal materials for scientific research and medicine on the one hand, is favorable for carrying out large-scale ecological breeding of medicinal animal centipedes in subsequent industrial pharmacy on the other hand, and is favorable for sustainable development of the industrial chain related to the medicinal animal centipedes.

Description

Reproduction method of centipedes hastellae

Technical Field

The invention belongs to the technical field of medicinal animal feeding, and particularly relates to a centipede hastelling breeding method.

Background

Centipedes, which are ancient members of terrestrial invertebrates and widely distributed in the world, are specialized as jaw limbs for first step feet, can secrete venom, and are mainly used for predation and defense. Centipede has been used as an animal medicine in China for more than two thousand years. As early as in Shen nong Ben Cao Jing, there is a record about the medication of centipedes, and the pharmacological actions of centipedes are classified as toxic, sweet and mild in flavor and entering the stomach meridian. The centipede animal medicine for Chinese medicine is prepared with centipede (Scolopendendrasubspinos mutilans L. Koch) as well as 6 species including Khaki centipede, ink river centipede, etc. as medicine centipede in different areas of China.

Scolopendra, a traditional Chinese medicine, has effects of counteracting toxic substances, killing parasite, dispelling pathogenic wind and relieving pain, and is commonly used for treating pyocutaneous disease, swelling toxin, acute mastitis, disease, intractable tinea, toothache, rheumatalgia, etc. Modern pharmacological researches find that main drug-effect substances of the centipedes comprise proteins, enzymes, fatty acids and the like, and have the effects of regulating lipid metabolism, reducing blood fat, enhancing coronary blood flow, improving immunity, resisting aging, tranquilizing and easing pain and the like. Modern medical research finds that centipedes can be used for treating various diseases, such as partial cancers, and centipede water extract has strong inhibition effect on ovarian cancer, liver cancer, lung cancer, gastric cancer, colon cancer and other cancer cells; and can be used for treating apoplexy, hemiplegia, infantile convulsion, etc., and male diseases with good therapeutic effect.

The centipede venom has more complex components and different species of centipede venom have different pharmacological properties. Moreover, compared with other toxic animal species, the centipede venom has more remarkable difference in components, which is also a material basis for long-standing medicine of important animals. After the venom of the centipede hastelling is purified, the venom protein or polypeptide substances of the centipede hastelling have various pharmacological activities different from other centipede species, such as trypsin inhibitory activity, platelet aggregation activity, voltage-gated potassium channel activity and the like. The pharmacological action of the differences also enables the centipede hastelling to have better advantages in the aspects of biological indexes and pharmacology.

Kyllinga cantonensis (Scolopendra subspinipes dehaani Brandt) belongs to the family Centipedes, is a centipede with large individual and large venom amount, generally has an individual body length of 7-20cm, and the average body length of adults fed in Xishuangbanna can reach 22cm. The centipedes hastelling are distributed in subtropical areas of tropical zones such as Yunnan, hainan, guangdong and Guangxi, but the quantity is small; investigation finds that the centipede Hardgrove is the dominant species in the West-Shuangbanna area of Yunnan. In Xishuangbanna, villous amomum species under rubber forest are distributed with more centipedes which are due to complex feeding habits and can prey on mollusks (such as snails) and various pests (such as locusts, grasshoppers, crickets, scarabs and the like). The centipede hastelleri has diversified application values, and from the perspective of traditional Chinese medicinal materials, the centipede hastelleri can be used as an important animal medicine resource; from the perspective of biological control of agricultural pests, the centipede hastelling can be further developed and utilized as the dominant natural enemy of rubber and fructus amomi planting areas, namely the centipede hastelling is utilized as a control agent of natural enemy organisms, and the pests are controlled by the principle of controlling the pests by the pests.

The centipede hastelling is one of the traditional Chinese medicinal materials in China and has good medicinal value. However, wild resources of the centipede hastelling are very limited at present, and the requirements of the medicinal market can not be met. Although the number of centipedes can be improved to a certain extent by artificial feeding, no report related to the artificial feeding technology of centipedes hastella is available so far. Due to the lack of key biological information and feeding technology for feeding the centipedes hasteng, the centipedes hasteng in the current market have different quality and are difficult to reach the standard of excellent centipedes, so that the standardized ecological process of the centipedes is blocked, and the safety and the effectiveness of clinical medication are difficult to ensure. Therefore, an efficient and scientific centipede breeding method is needed to be developed to improve the breeding quality of centipedes hastelling and promote the standardized and large-scale breeding process of the centipedes hastelling to meet the standard of standard medicine use. The patent researches the breeding technology of the centipede hastelling for the first time, and has important significance for further development and utilization of the centipede hastelling.

Disclosure of Invention

The invention provides a reproduction method of centipedes hastelling for the first time, which is simple to operate, has easily obtained reproduction tools, saves the feeding cost, obviously improves the survival rate of the centipedes, greatly shortens the medicament application period of the centipedes, reduces the raw material cost of the centipedes, can produce a large amount of centipedes hastelling with consistent quality and excellent quality as traditional Chinese medicinal materials in a large scale in a short time for supplying to the market, and provides medicinal centipede raw materials with controllable quality.

In order to achieve the purpose, the technical scheme adopted by the invention is as follows:

a reproduction method of centipedes hastelling comprises the following steps:

s1, culturing the centipedes hastelling to an oviposition stage by using a climatic chamber, picking out female centipedes in the oviposition stage after one week of oviposition, controlling the temperature in the climatic chamber to be 18-34 ℃, controlling the humidity to be 60-80% and controlling the light cycle to be dark: illumination =12:12;

s2, respectively transferring the picked brooding Khattai centipedes to a feeding box for independent feeding, placing each feeding box in a climatic chamber, paving padding materials with the height of 1.5-2.5cm in each feeding box, wherein the padding materials are prepared from coconut chaff and brick red soil according to the proportion of 7: (2-4), 15-25mL of traditional Chinese medicine liquid is added into the padding, the humidity of the padding is adjusted to be 20% -28% by water, and the traditional Chinese medicine liquid is prepared from Dai Baijie and clerodendranthus spicatus;

s3, transferring the brood Kjellyfish centipedes which have the eggs to a climatic box to be separately fed with the larva after the brood Kjellyfish centipedes are hatched, transferring the brood Kjellyfish centipedes from the feeding box to the climatic box to be fed when the brood Kjellyfish centipedes can be independently moved, and feeding the brood Kjellyfish centipedes to an adult;

in the feeding period of the steps S1 to S3, 7-9g of feed is fed to the centipedes each time in the non-oviposition period, the feed fed to the centipedes each time in the non-oviposition period is increased to 10-12g, the centipedes are not fed any more in the hatching period, the centipedes are observed and fed once every 3 days, and the centipedes are fed 20:00, and feeding feed comprises living animals and common meat.

The established breeding method of the centipede hastelling can accurately and simply carry out propagation, and provide the centipede Chinese medicinal materials which have excellent biological indexes, more consistent growth vigor, active and robust individuals and good quality and can meet the first-level standard requirements of the Chinese medicinal materials. Is beneficial to large-scale ecological breeding of the centipedes hastelling, is beneficial to promoting the sustainable development of related industrial chains, and is beneficial to promoting the rural happy strategy practice taking traditional Chinese medicine breeding as a gripper. Therefore, the biological characteristics of the centipedes susceptible to fungal diseases are combined, the feeding boxes are adopted for feeding separately in the incubation period and the larva growth period, diversified living animal characteristic feeds are fed in the feeding period, and the characteristic Dai medicine is adopted as an additive component of the mixture padding, so that the cross infection of diseases can be effectively prevented, the centipedes hasten are enabled to be good in performance on various morphological indexes, the centipedes are enabled to be more robust, and the breeding of good varieties is facilitated; the centipede medicinal parts are the whole centipede, the centipede hastelling offspring bred by the breeding method has excellent biological indexes, more consistent growth vigor, active and robust individuals, 80 larvae can be produced by each female, the average length of the adult is 22.1cm, the quality is good, and the centipede hastelling offspring can meet the first-level standard requirements of traditional Chinese medicinal materials. The yield and the quality of the centipede medicinal materials are obviously improved, and the cost such as the time period is effectively reduced.

The method provided by the invention has important significance in breeding and standardized large-scale feeding of the centipedes Harden, gradually meeting the resource requirements of domestic medicine enterprises, promoting the ordered development of related industrial chains of the centipedes Chinese medicinal materials and solving the market requirements of the domestic centipedes Chinese medicinal materials.

The survival rate of the centipedes hastelling can be effectively improved by adopting the optimal temperature illumination humidity condition. Preferably, the temperature of the climatic chamber is 22-30 ℃.

Preferably, the preparation method of the traditional Chinese medicine liquid comprises the following steps: uniformly mixing Daibaijie and folium orthosiphoni according to the weight ratio of 1:2, adding water, boiling, decocting to one fourth of the original volume, and filtering dregs to obtain the traditional Chinese medicine liquid. Further, the feed-liquid ratio of the Daibeibaijie to water is 3g:400mL.

Preferably, in the feed for feeding, the mass ratio of the living animal to the common meat is 1:1.

Further, the living animals are any one of crickets, locusts, grasshoppers, yellow mealworms and snails, the common meat is any one of fresh chicken, chicken livers and pig livers, and the living animals and the common meat are matched at will and are fed and changed with different matching combinations each time.

Preferably, the bedding is prepared from coconut coir and red loam according to the weight ratio of 7:3 in a weight ratio.

Preferably, the laying height of the padding is 2.0cm.

Preferably, 30-60 centipedes are raised in each climatic chamber.

Preferably, 20-40 rearing boxes are placed per climatic chamber.

Preferably, during the rearing in steps S1 to S3, the climatic chamber is kept quiet, without excessive disturbance other than the daily rearing management.

Compared with the prior art, the invention has the beneficial effects that:

the invention provides a breeding method of centipedes hastelling, which is simple to operate, simple and easy to operate in breeding steps, easy to obtain breeding tools, not only saves the breeding cost, but also can effectively shorten the breeding period, obviously improve the survival rate and the sub-generation quantity of the centipedes hastelling and reduce the fatality rate; meanwhile, the centipede hastelling bred by the method has excellent offspring biological indexes, more consistent growth vigor, active and robust individuals and good quality, can meet the primary standard requirements of traditional Chinese medicinal materials, can provide stable and reliable animal materials for scientific research and medicine on the one hand, is favorable for carrying out large-scale ecological breeding of medicinal animal centipedes in subsequent industrial pharmacy, and is favorable for sustainable development of the industrial chain related to the medicinal animal centipedes on the other hand.

Drawings

FIG. 1 is a climatic chamber for propagation;

FIG. 2 shows a breeding box;

FIG. 3 is a view of Khatherum centipede raised in a climatic chamber;

FIG. 4 is a partial Khatherum centipede in a rearing box;

fig. 5 is an oviferous female centipede hastella.

Detailed Description

The following further describes the embodiments of the present invention. It should be noted that the description of the embodiments is provided to help understanding of the present invention, but the present invention is not limited thereto. In addition, the technical features involved in the embodiments of the present invention described below may be combined with each other as long as they do not conflict with each other.

The experimental procedures in the following examples were carried out by conventional methods unless otherwise specified, and the test materials used in the following examples were commercially available by conventional methods unless otherwise specified.

Example 1 establishment of reproduction method of Khattachia

(1) Early preparation:

1) The artificial propagation tools such as a climatic box (purchased from Ningbo Saifu laboratory instruments Co., ltd., an intelligent artificial climatic box with the model of PRX-250B), a feeding box (with the length multiplied by the width multiplied by the height multiplied by 19cm multiplied by 12.5cm multiplied by 7.5cm, 10-15 small holes with the diameter of 2mm are respectively arranged on two long sides, purchased from Huangyan palm Ching spider pet products Co., ltd., taizhou city) and tweezers need to be disinfected by 75% alcohol before use.

2) Preparation of traditional Chinese medicine liquid for padding (taking 500mL as an example): the traditional Chinese medicine liquid is prepared from Dai medicine, the Dai medicine comprises Dai Baijie (Marsdenia tenacissima) and kidney tea (Cleriodendanthus spicatus), 15g of the Dai Baijie and 30g of the kidney tea are respectively weighed, 2000mL of water is added after uniform mixing, the mixture is boiled to 500mL after boiling, the medicine residue is filtered, the traditional Chinese medicine liquid is prepared, and the ice box is placed for standby after normal temperature cooling.

3) Feed for feeding: comprises a living animal and common meat, wherein the mass ratio of the living animal to the common meat is 1:1; the living animals are any one of crickets, locusts, grasshoppers, yellow mealworms and snails, the common meat is any one of fresh chicken, chicken livers and pig livers, and the living animals and the common meat are matched at will and are fed and changed with different matching combinations each time. For example, the first feeding is carried out by matching and combining cricket and pork liver, the second feeding is changed into locust and chicken, and the third feeding is changed into grasshopper and chicken liver … …, namely, the components matched in two adjacent times are different, and the rest is done in the same way.

(2) The specific propagation process comprises the following steps:

1) Taking a climatic chamber as a feeding chamber (as shown in figure 1), culturing the centipedes with Ha's style to an oviposition stage, picking out female centipedes at the oviposition stage after one week of oviposition, feeding 30-60 (male and female centipedes in mixed culture) in each climatic chamber, controlling the temperature in the climatic chamber to be 18-34 ℃, the humidity to be 60-80% and the light cycle to be dark: illumination =12:12, ensuring that the climate box is in a feeding room with good natural ventilation;

2) Respectively transferring the picked oviposition centipedes hastelling with tweezers to feeding boxes (shown in figure 2) for independent feeding, placing the feeding boxes in climate boxes, wherein each climate box is provided with 20-40 feeding boxes, and padding with the height of 1.5-2.5cm is paved in each feeding box, and is prepared from coconut chaff and brick red soil according to the ratio of 7: (2-4), adding 15-25mL of traditional Chinese medicine liquid into the padding, and adjusting the humidity of the padding to be 20% -28% by using sterile water;

3) After brooding Khattacharbe is hatched into young Khattacharbe, the female Khattacharbe is transferred from the feeding box to the climate box by using tweezers to be fed separately from the larva, the young Khattacharbe is transferred from the feeding box to the climate box when being cultured to the stage of independent activity, and the feeding is finished after the young Khattacharbe is fed to the adult.

During the feeding period of each feeding step, keeping the climate box quiet, not interfering too much except daily feeding management, feeding 7-9g of feed for each time in a non-oviposit period, properly increasing the amount of the feed during the oviposit period, increasing the amount of the feed for each time to 10-12g, not feeding the centipedes (feeding the centipedes according to normal standard after the centipedes are hatched out of larvae), observing and feeding once every 3 days, and feeding 20:00 is carried out.

Example 2 research on growth and development indexes of centipede Hardgrove at different development stages

The centipede hastelling is artificially fed according to the centipede hastelling propagation method in the embodiment 1, and the feeding process is briefly introduced as follows:

(1) Culturing the centipedes in a climatic chamber until the centipedes are in an oviposition stage (the centipedes raised in the climatic chamber are shown in figure 3; the female centipedes in the oviposition are shown in figure 5), wherein the temperature in the climatic chamber is 26 +/-1 ℃, the humidity is 70 +/-5%, and the light cycle is dark: illumination =12:12; then picking 20 female centipedes which have spawned in the same batch;

(2) Feeding 20 female centipedes with eggs in a feeding box (the Khatherd centipedes in the feeding box are shown in figure 4) separately, laying a layer of padding with the thickness of 2cm (the weight ratio of coconut chaff to brick red soil is 7:3) in the box, adding 20mL of traditional Chinese medicine liquid into the padding, and adding water to adjust the humidity to 24%;

keeping a climate box quiet during feeding, keeping the climate box quiet during the feeding period, not interfering with daily feeding management, feeding the centipedes about 8g each time in a non-oozing period (the first feeding is crickets: pork liver = 1:1), properly increasing the feeding amount during the oozing period, increasing the feeding amount each time to about 11g, not feeding the centipedes any more in an incubation period (feeding the centipedes according to normal standards after the centipedes hatch the larvae), observing and feeding the centipedes once every 3 days, and feeding the centipedes 20:00 is carried out;

(3) After hatching the young centipedes, the brooding centipedes are transferred to a climatic box to be fed separately from the larvae, and the young centipedes are transferred to the climatic box to be fed when being cultured until the centipedes can move independently (about 45 days are needed) until being fed to adults (about one year is needed).

In the raising process, growth and development indexes of offspring eggs, larvae, sub-adults and adults in each development stage are respectively observed, 20 centipedes are observed in each treatment group, observation is carried out for 2-3 times every week, morphological characteristics of each development stage are observed by using a stereoscopic microscope, and data such as the body length of the centipedes are measured by using a ruler.

As can be seen from Table 1, the centipedes Harris bred by the centipede breeding method of example 1 have an average adult length of 22.1cm. According to the reported standards of centipede medicinal materials (refer to a document 'Hu Chaoyi, and the like; the research on the commodity specification grade standard of centipede medicinal materials [ J ]. Anhui agricultural science, 2019,47 (16): 209-211, 253', in particular to the content of the commodity specification grade division of centipedes in the table 5 in the document), the first-level standard is that the body length is more than or equal to 14cm, so that the centipedes hastening fed by the method can reach the standard of the first-level medicinal materials.

TABLE 1 biological observations of the different developmental stages of Khatherum centipede

Note: data in the table are mean ± sem.

EXAMPLE 3 Effect of different temperatures on the growth and development of Khatherum

Culturing the centipedes hastelling by using a climatic chamber to the oviposition stage, selecting the female centipedes with the same batch of oviposition for testing, artificially feeding the centipedes hastelling according to the breeding method of the centipedes hastelling in the embodiment 1, observing 20 centipedes in each treatment group for 2-3 times in each week, wherein the feeding specific parameters are the same as those in the embodiment 2, and simultaneously, the climatic chamber temperature of each treatment group is respectively set to be 18 ℃,22 ℃, 26 ℃, 30 ℃ and 34 ℃; the development stages of the offspring at each development stage were observed, and the stage (unit d) and death at each development stage were recorded for 5 temperature treatments.

As can be seen from Table 2, the overall trend of the development history of the Khatherum centipede is gradually shortened along with the increase of the temperature, wherein the mortality rates of the centipedes treated at 18 ℃ and 34 ℃ are obviously higher than those of the centipedes treated at 22 ℃, 26 ℃ and 30 ℃, and the mortality rates of the centipedes treated at 22-30 ℃ can be 0 in all development stages, so that the temperature of 22-30 ℃ is the optimum feeding temperature.

TABLE 2 duration of various developmental stages of Khatherum centipede at different temperatures

Note: data in the table are mean ± sem.

Example 4 study of the propagation time of Kyllinga brevifolia

Culturing the centipedes hastelling by a climatic chamber to the oviposition stage, selecting the female centipedes with eggs of the same batch for testing, dividing the centipedes into two equal parts with 20 heads each, wherein one part is artificially fed according to the breeding method of the centipedes hastelling in the embodiment 1, the specific feeding parameters are the same as those in the embodiment 2, and the temperature of the climatic chamber is 26 ℃. The observation is performed 2-3 times per week, and the development period of each development stage of ovum, larva, sub-adult and adult is recorded.

And the other part is placed in a rubber forest planting area of the Dai autonomous State of the Xishuangbanna, yunnan province for field propagation, investigation is carried out once every half month, system investigation is continuously carried out, and development periods of development stages of eggs, larvae, sub-adults and adults are observed and recorded.

The development period of indoor feeding is shown in table 2, the development time of the centipede hastelling from an egg to an adult is 348.5 days; the development period of the field propagation is shown in Table 3, and the development time of the Khatherum polycephalum from egg to adult is 592.4d; compared with the field, the growth duration of indoor feeding is accelerated by 243.9d. Therefore, the centipede fed by the method can obviously shorten the development period of the centipede and improve the breeding efficiency of the centipede.

TABLE 3 growth history of wild centipedes

Stage of development	History of life	Development calendar period (d)
			Egg	From first 1 month to first 2 months	17.5±3.2
Development of larvae to independent activities	From 1 late ten days to 2 late ten days	35.1±4.8
			From larval to sub-adult	Last ten days of 3 months to middle ten days of June	425.4±24.7
From juvenile to adult	7 months and early in the next year	574.9±26.3

Note: data in the table are mean ± sem.

Example 5 Khatag centipede 1 generation quantity study

After the centipedes hastelling are cultured to the egg-carrying stage by using a climatic chamber, 15 female centipedes which carry eggs in the same batch are selected for testing, artificial feeding is carried out according to the breeding method of the centipedes hastelling in the embodiment 1, and the specific feeding parameters are the same as those in the embodiment 2. The number of offspring (F1 generation) was recorded separately, 2-3 times per week. Meanwhile, 3 female centipedes which have spawned are collected and observed in the field as field control, and the number of filial generations (F1 generation) of the female centipedes is observed.

As shown in the statistical results of the F1 generation numbers of the Khattacharbe in Table 4, the average number of the F1 generation of the Khattacharbe bred by the method is 80 heads/female, and the average number of the field control group is 51 heads/female. Therefore, the centipede fed by the method can greatly shorten the period of application of the centipede as a medicine, and effectively reduce the time period and other costs.

TABLE 4 statistics of the number of generations of Khattachar F1

Note: data in the table are mean ± sem.

The embodiments of the present invention have been described in detail, but the present invention is not limited to the described embodiments. It will be apparent to those skilled in the art that various changes, modifications, substitutions and alterations can be made in the embodiments without departing from the principles and spirit of the invention, and these embodiments are still within the scope of the invention.

Claims

1. The breeding method of the centipede hastelleri is characterized by comprising the following steps:

s2, respectively transferring the picked oviposition Khatsuki centipedes into feeding boxes for independent feeding, placing the feeding boxes into a climate box, and paving 1.5-2.5cm high padding materials in the feeding boxes, wherein the padding materials are prepared from coconut coir and brick red soil according to the weight ratio of 7: (2-4), adding 15-25mL of Chinese medicinal liquid into the padding, adjusting the humidity of the padding to 20% -28% by using water, mixing Dai Baiji and kidney tea uniformly according to the weight ratio of 1:2, adding water, boiling, decocting to one fourth of the original volume, and filtering dregs to obtain the Chinese medicinal liquid;

during the raising period of the steps S1 to S3, 7-9g of feed is fed to the centipedes every time in the non-egg-laying period, the feed fed every time in the egg-laying period is increased to 10-12g, the centipedes are not fed any more in the hatching period, the centipedes are observed and fed once every 3 days, the feed for feeding comprises living animals and common meat, the living animals are any one of crickets, locusts, mealworms and snails, the common meat is any one of fresh chicken, chicken livers and pork livers, and the living animals and the common meat are matched at will and different matching combinations are changed every time of feeding.

2. The breeding method of centipedes hastelli according to claim 1, wherein the temperature of the climatic chamber is 22-30 ℃.

3. The breeding method of centipedes harveyi according to claim 1, wherein the mass ratio of the living animals to the common meat in the feed is 1:1.

4. The breeding method of centipedes hastella according to claim 1, wherein the padding is prepared from coconut coir and brick red soil according to the weight ratio of 7:3 in a weight ratio.

5. The breeding method of centipedes hastella according to claim 1, wherein the laying height of the padding is 2.0cm.

6. The breeding method of centipedes hastelli according to claim 1, wherein 30-60 centipedes are bred in each climatic chamber.

7. The breeding method of centipedes hastelling as claimed in claim 1, wherein 20-40 rearing boxes are placed in each climate box.

8. The breeding method of centipedes harveyi according to claim 1, wherein during the breeding period of steps S1 to S3, the climate box is kept quiet without excessive disturbance except for daily breeding management.