CN113455470B - Method for artificially culturing toad feeding insects - Google Patents
Method for artificially culturing toad feeding insects Download PDFInfo
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Images
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; CARE OF BIRDS, FISHES, INSECTS; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New breeds of animals
- A01K67/033—Rearing or breeding invertebrates; New breeds of invertebrates
Abstract
The invention discloses a method for artificially cultivating toads, which comprises the steps of cleaning toads by using extract liquid of mangnolia officinalis, immature bitter orange, mirabilite and rheum officinale, pulping the toads, adding a selenious acid triglyceride mixture and honey, placing a maggot cultivating box for chrysomyia megacephala to eat and grow maggots, promoting maggot production by using porous microspheres containing indole, giving a proper growth temperature to the maggots by adjusting the light, humidity and temperature in the maggot cultivating box, passing through water of 48-50 ℃, and naturally airing to obtain the toads; the cultured toad feeding insects have high utilization rate of effective active ingredients in toads, high selenium content and strong inhibition activity on malignant tumors; the yield is high, the quality is uniform, and the culture period is short; the method is simple and convenient, has no regional limitation, can ensure the stability of maggot raising conditions, and is suitable for long-term mass production.
Description
Technical Field
The invention relates to the field of toad feeding, in particular to a method for artificially culturing toad feeding.
Background
The toad maggot is a kind of maggot larva formed by laying fly on the dead body of toad and eating the maggot sprout out of toad meat, and is a traditional and precious animal in China. From ancient times to the present, a sentence of medicine proverb was introduced in the medical field: bufo frog can not produce maggot, and the produced maggot is treasure. Because the toad maggot has magical pharmacological action, it can cure several difficult and complicated diseases. As early as over 300 years ago, Li Shizhen has detailed and definite record on the unique medicinal value and magical curative effect of toad maggots in Ben Cao gang mu (Vol. fortieth).
The toad eating maggot is rich in a plurality of effective substances of the toads and the maggots, integrates the effects of the toads and the maggots, and actually takes fly maggots as carriers to selectively absorb and enrich effective components on the toads through the fly maggots. The Bufonis venenum mainly contains Bufonis venenum, chitin, natural agglutinin, various amino acids, protein, SOD, minerals, and bufalin, and has effects of tonifying heart, promoting urination, relieving pain, diminishing inflammation, and inhibiting tumor growth.
The function of the toad venom is very wide, but the toad venom has certain toxicity and is easy to be overdosed after being directly taken, so that the safety problem exists. The toad feeding is relatively safe, has the efficacy of toad venom and the function and efficacy of maggots, is a brand new compound product, and has been researched a lot.
Traditional toad eating insects are mainly naturally formed by wild accidentally dead toads, the toads eating insects collected in the past are bred by dead toads which are killed by livestock, bitten by wild animals and putrescent toads dead bodies after being killed by people, the toads eating insects are small in amount, can meet but cannot be obtained, are difficult to obtain stably and cannot meet a large number of market demands, patent CN1056956C discloses a method for artificially breeding the toad eating insects, the method is influenced by natural conditions, uncertain factors are more, the yield is limited, dirty matters in the toads are not removed, patent CN 104585 discloses a breeding method for the toad eating rejuvenation insects and prepared medicine raw materials, the breeding method is complicated, and the content of components in the toads eating insects is not uniform.
Disclosure of Invention
In view of the above situation, on the basis of the previous method, the artificial cultivation method of the toad feeding insects is provided, and the toad feeding insects with strong pharmacological activity and uniform component content can be quickly, conveniently, continuously and stably cultured in an industrialized mode.
The technical proposal for solving the problem is that,
an artificial cultivation method of toad feeding insects comprises the following steps:
s1, cleaning: cleaning living toads with clear water; then putting the toad into the mixed solution for 72 h;
s2, pulping: taking out the toad in S1, washing with flowing clear water for 20-24h, taking out the toad, beating the toad to cause dizziness, then pulping the toad uniformly, adding selenious acid triglyceride mixture and honey, and pulping the mixture uniformly again to obtain mixed toad pulp;
s3, maggot breeding: pouring the mixed toad slurry into a maggot raising pool in a maggot raising box, putting the porous microspheres into a grid box, putting chrysomyia megacephala into a maggot laying chamber, adjusting light, humidity and temperature, starting a ventilation device, starting a magnetic stirring device, continuously eating the mixed toad slurry by the chrysomyia megacephala and performing generation propagation, and obtaining the maximum toad eating number on the fifth day;
s4, maggot collection: turning off the light in the maggot laying chamber, turning on the light in the light chamber, allowing the toad to climb into the maggot collecting chamber through the pore channel, turning on a water sprayer in the maggot collecting chamber, after 2-3h, allowing the toad to climb in, and cleaning to obtain clean fresh maggots;
s5, putting the fresh maggots into water, heating to 48-50 ℃, filtering to remove water after 5-10min, and naturally airing for 2-3 days to obtain the toad eating maggots.
Further, the porous microspheres are porous activated carbon soaked in indole.
Further, the maggot raising box comprises a maggot producing chamber and a maggot collecting chamber, a maggot raising pool is arranged at the bottom of the maggot producing chamber, a plurality of grid boxes are arranged outside the maggot raising pool, pore passages are arranged outside the grid boxes, and the pore passages are communicated with the maggot producing chamber and the maggot collecting chamber.
Further, the preparation method of the porous microsphere comprises the following steps: firstly, indole is dissolved in ethanol to prepare the indole with the concentration of 1 × 10-4And (3) pouring the g/mL indole solution into porous activated carbon, immersing, stirring for 30-60min, and drying at 60-70 ℃ for 6-8h to obtain the porous microspheres.
Further, the maggot raising case comprises a maggot producing chamber and a maggot collecting chamber, the maggot producing chamber is arranged above the maggot collecting chamber, case shells are arranged outside the maggot producing chamber and the maggot collecting chamber, the case shells are black, the top of the maggot producing chamber is provided with an adjustable light, humidity detector, spraying device and air exchange device, a maggot raising pool is arranged at the bottom of the maggot producing chamber, magnetic balls are arranged in the maggot raising pool, a magnetic stirrer is arranged below the maggot raising pool, a plurality of net boxes are arranged outside the maggot raising pool, pore passages are arranged outside the net boxes and are communicated with the maggot producing chamber and the maggot collecting chamber, the lower edge of the pore passages is provided with an extension plate which extends outwards, the top of the maggot collecting chamber is provided with a water sprayer, light chambers are arranged at two sides of the maggot collecting chamber, a maggot blocking partition plate between the light chamber and the maggot collecting chamber is made of transparent glass, the maggot collecting box is arranged in the middle of the maggot collecting chamber, the cover-free position above the maggot collecting box, the transparent glass is made of transparent glass, a plurality of net holes are uniformly distributed on the maggot collecting box, the bottom of the maggot collecting chamber is provided with a water outlet.
Further, the preparation method of the mixed solution comprises the following steps: 100g of magnolia officinalis, 100g of immature bitter orange, 100g of mirabilite and 100g of rhubarb are taken, mixed and crushed, decocted in water for 30min and filtered, 25-30 kg of clean water is added into the filtrate, and the water temperature is kept at 26-28 ℃.
Further, the lamplight of the maggot producing chamber and the lamplight chamber is variable, before the toad pulp is put in, ultraviolet light is firstly used for irradiating for 5-10min, sterilization is carried out, after the toad pulp is put in, white light irradiation is carried out for 6:00-20:00, then the lamplight is turned off in 20:00-6:00 time period, the humidity is controlled to be 75% +/-5%, and the temperature is controlled to be 28 +/-2 ℃; the light of the light chamber is white light when maggots are collected.
Further, the mass quantity ratio of the mixed toad pulp to the chrysomyia megacephala is 1000g/200, wherein the quantity ratio of male and female of the chrysomyia megacephala is 1: 1; the female chrysomyia megacephala is to-be-born chrysomyia megacephala.
Furthermore, after maggots are collected, chrysomyia megacephala imagoes in the maggot breeding box can be continuously used for breeding toads in the next round.
Furthermore, the selenious acid triglyceride mixture is synthesized by esterification of sunflower oil and selenious acid, the content of the selenious acid triglyceride is 2%, and the selenious acid triglyceride mixture mainly has a dioxaselenolane structure.
Further, the mass volume ratio of the toad, selenious acid triglyceride mixture and honey is as follows: 800-1000g, 20-24g, 50-60 g.
Further, the toad variety is a Chinese toad.
Due to the adoption of the technical scheme, compared with the prior art, the invention has the following advantages;
the toad larvae cultured by the method for artificially culturing the toad larvae has high utilization rate of effective active ingredients in the toads, high selenium content and stronger inhibitory activity on malignant tumors; the yield is high, the quality is uniform, and the culture period is short; the method is simple and convenient, has no regional limitation, can ensure the stability of maggot raising conditions, and is suitable for long-term mass production.
Drawings
FIG. 1 is a structural view of a maggot raising box according to the present invention;
FIG. 2 is a sectional view of the maggot raising box of the present invention.
Detailed Description
The foregoing and other technical and scientific aspects, features and utilities of the present invention will be apparent from the following detailed description of the embodiments, which is to be read in connection with the accompanying drawings of fig. 1-2. The structural contents mentioned in the following embodiments are all referred to the attached drawings of the specification.
Example 1
An artificial cultivation method of toad feeding insects comprises the following steps:
s1, cleaning: washing 16kg of living toad with clear water; then putting the toad into the mixed solution for 72 h;
s2, pulping: taking out the toads in the S1, washing the toads with flowing clear water for 20h, taking out the toads, beating the toads to cause dizziness, then pulping the toads uniformly, adding 40g of selenious acid triglyceride mixture and 100g of honey, and pulping the toads uniformly again to obtain mixed toad pulp;
s3, maggot breeding: pouring the mixed toad slurry into a maggot raising pool in a maggot raising box, putting the porous microspheres into a grid box 15, putting chrysomyia megacephala into a maggot laying chamber, adjusting light, humidity and temperature, starting a ventilation device, starting a magnetic stirring device, continuously eating the mixed toad slurry by the chrysomyia megacephala and performing generation and propagation, and obtaining the maximum number of toads eating on the fifth day;
s4, maggot collection: turning off the light in the maggot laying chamber, turning on the light in the light chamber, allowing the toad to climb into the maggot collecting chamber through the pore channel, turning on a water sprayer in the maggot collecting chamber, after 2h, allowing the toad to climb in, and cleaning to obtain clean fresh and alive maggots;
s5, putting the fresh maggots into water, heating to 48 ℃, filtering to remove water after 5min, and naturally airing for 2 days to obtain the toad food.
After maggots are collected, chrysomyia megacephala imagoes in the maggot breeding box can be continuously used for breeding toads in the next round.
The preparation method of the porous microsphere comprises the following steps: firstly, indole is dissolved in ethanol to prepare the indole with the concentration of 1 × 10-4And (3) pouring the indole solution of g/mL into porous activated carbon, immersing, stirring for 30min, and drying for 6h at 60 ℃ to obtain the porous microspheres.
As shown in figure 1 and figure 2, the maggot raising case comprises a maggot producing chamber and a maggot collecting chamber, wherein the maggot producing chamber is arranged above the maggot collecting chamber, a case shell 1 is arranged outside the maggot producing chamber and the maggot collecting chamber, the case shell 1 is black, the top of the maggot producing chamber is provided with adjustable light 10, a humidity detector 20, a spraying device 12 and a ventilation device 11, a maggot raising pool 13 is arranged at the bottom of the maggot producing chamber, magnetic balls 19 are arranged in the maggot raising pool 13, a magnetic stirrer 18 is arranged below the maggot raising pool, a plurality of net boxes 15 are arranged outside the maggot raising pool, a pore passage 16 is arranged outside the net boxes 15, the pore passage 16 is communicated with the maggot producing chamber and the maggot collecting chamber, an extension plate 17 is arranged at the lower edge of the pore passage 16, the extension plate 17 extends outwards, a water sprayer 30 is arranged at the top of the maggot collecting chamber, light chambers 31 are arranged at two sides of the maggot collecting chamber, a baffle plate between the light chamber 31 and the maggot collecting chamber is made of transparent glass, a collection box 32 is arranged in the middle of the maggot collecting chamber, the collection box is made of transparent glass, a plurality of net holes are uniformly distributed on the maggot collecting box 32, the net holes do not allow the smallest maggots to pass through, and a water outlet 33 is arranged at the bottom of the maggot collecting chamber.
The preparation method of the mixed solution comprises the following steps: 100g of magnolia officinalis, 100g of immature bitter orange, 100g of mirabilite and 100g of rhubarb are taken, mixed and crushed, decocted in water for 30min and filtered, 25 kg of clear water is added into the filtrate, and the water temperature is kept at 26 ℃.
Before the toad slurry is put in, the lamp of a maggot breeding box is firstly adjusted to be ultraviolet light for irradiating and sterilizing for 5min, after the toad slurry is put in, white light is irradiated for 6:00-20:00, then the lamp light is turned off in 20:00-6:00 time period, the humidity is controlled at 70%, the temperature is controlled at 26 ℃, and the rotating speed of magnetic stirring is 200 r/min.
The mass quantity ratio of the mixed toad slurry to the chrysomyia megacephala is 1000g/200, wherein the quantity ratio of male to female of the chrysomyia megacephala is 1: 1; the female chrysomyia megacephala is to-be-born chrysomyia megacephala.
The selenious acid triglyceride mixture is synthesized by esterification of sunflower oil and selenious acid, the content of the selenious acid triglyceride is 2%, and the selenious acid triglyceride mixture mainly has a dioxaselenolane structure.
The toad variety is Bufo siccus.
Example 2
An artificial cultivation method of toad feeding insects comprises the following steps:
s1, cleaning: washing 18kg of living toad with clear water; then putting the toad into the mixed solution for 72 h;
s2, pulping: taking out the toads in the S1, washing the toads with flowing clear water for 22h, taking out the toads, beating the toads to cause dizziness, then pulping the toads uniformly, adding 44g of selenious acid triglyceride mixture and 110g of honey, and pulping the toads uniformly again to obtain mixed toad pulp;
s3, maggot breeding: pouring the mixed toad slurry into a maggot raising pool in a maggot raising box, putting the porous microspheres into a grid box, putting chrysomyia megacephala into a maggot laying chamber, adjusting light, humidity and temperature, starting a ventilation device, starting a magnetic stirring device, continuously eating the mixed toad slurry by the chrysomyia megacephala and performing generation propagation, and obtaining the maximum toad eating number on the fifth day;
s4, maggot collection: turning off the light in the maggot laying chamber, turning on the light in the light chamber, allowing the toad to climb into the maggot collecting chamber through the pore channel, turning on a water sprayer in the maggot collecting chamber, after 2.5h, allowing the toad to climb in, and cleaning to obtain clean fresh maggots;
s5, putting the fresh maggots into water, heating to 49 ℃, filtering to remove water after 8min, and naturally airing for 3 days to obtain the toad food.
After maggots are collected, chrysomyia megacephala imagoes in the maggot breeding box can be continuously used for breeding toads in the next round.
The preparation method of the porous microsphere comprises the following steps: firstly, indole is dissolved in ethanol to prepare the indole with the concentration of 1 × 10-4And (3) pouring the indole solution of g/mL into porous activated carbon, immersing, stirring for 45min, and drying for 7h at 65 ℃ to obtain the porous microspheres.
The preparation method of the mixed solution comprises the following steps: 100g of magnolia officinalis, 100g of immature bitter orange, 100g of mirabilite and 100g of rhubarb are taken, mixed and crushed, decocted in water for 30min and filtered, 28 kg of clear water is added into the filtrate, and the water temperature is kept at 27 ℃.
Before the toad slurry is put in, the lamp of a maggot breeding box is firstly adjusted to be ultraviolet light for irradiating and sterilizing for 8min, after the toad slurry is put in, white light is irradiated for 6:00-20:00, then the lamp light is turned off in 20:00-6:00 time period, the humidity is controlled at 75%, the temperature is controlled at 28, and the rotating speed of magnetic stirring is 250 r/min.
The mass quantity ratio of the mixed toad slurry to the chrysomyia megacephala is 1000g/200, wherein the quantity ratio of male to female of the chrysomyia megacephala is 1: 1; the female chrysomyia megacephala is to-be-born chrysomyia megacephala.
The selenious acid triglyceride mixture is synthesized by esterification of sunflower oil and selenious acid, the content of the selenious acid triglyceride is 2%, and the selenious acid triglyceride mixture mainly has a dioxaselenolane structure.
The toad variety is Bufo siccus.
Example 3
An artificial cultivation method of toad feeding insects comprises the following steps:
s1, cleaning: washing 20kg of living toad with clear water; then putting the toad into the mixed solution for 72 h;
s2, pulping: taking out the toads in the S1, washing the toads with flowing clear water for 24h, taking out the toads, beating the toads to cause dizziness, then pulping the toads uniformly, adding 48g of selenious acid triglyceride mixture and 120g of honey, and pulping the toads uniformly again to obtain mixed toad pulp;
s3, maggot breeding: pouring the mixed toad slurry into a maggot raising pool in a maggot raising box, putting the porous microspheres into a grid box, putting chrysomyia megacephala into a maggot laying chamber, adjusting light, humidity and temperature, starting a ventilation device, starting a magnetic stirring device, continuously eating the mixed toad slurry by the chrysomyia megacephala and performing generation propagation, and obtaining the maximum toad eating number on the fifth day;
s4, maggot collection: turning off the light in the maggot laying chamber, turning on the light in the light chamber, allowing the toad to climb into the maggot collecting chamber through the pore channel, turning on a water sprayer in the maggot collecting chamber, after 3h, allowing the toad to climb in, and cleaning to obtain clean fresh and alive maggots;
s5, putting the fresh maggots into water, heating to 50 ℃, filtering to remove water after 10min, and naturally airing for 3 days to obtain the toad food.
After maggots are collected, chrysomyia megacephala imagoes in the maggot breeding box can be continuously used for breeding toads in the next round.
The preparation method of the porous microsphere comprises the following steps: firstly, indole is dissolved in ethanol to prepare the indole with the concentration of 1 × 10-4And (3) pouring the indole solution of g/mL into porous activated carbon, immersing, stirring for 60min, and drying for 8h at 70 ℃ to obtain the porous microspheres.
The preparation method of the mixed solution comprises the following steps: 100g of magnolia officinalis, 100g of immature bitter orange, 100g of mirabilite and 100g of rhubarb are taken, mixed and crushed, decocted in water for 30min and filtered, 30 kg of clear water is added into the filtrate, and the water temperature is kept at 28 ℃.
Before the toad slurry is put in, the lamp of a maggot breeding box is adjusted to be ultraviolet light to irradiate and sterilize for 5-10min, after the toad slurry is put in, white light irradiation is carried out for 6:00-20:00, then the lamp light is turned off in 20:00-6:00 time period, the humidity is controlled at 80%, the temperature is controlled at 30 ℃, and the rotating speed of magnetic stirring is 300 r/min.
The mass quantity ratio of the mixed toad slurry to the chrysomyia megacephala is 1000g/200, wherein the quantity ratio of male to female of the chrysomyia megacephala is 1: 1; the female chrysomyia megacephala is to-be-born chrysomyia megacephala.
The selenious acid triglyceride mixture is synthesized by esterification of sunflower oil and selenious acid, the content of the selenious acid triglyceride is 2%, and the selenious acid triglyceride mixture mainly has a dioxaselenolane structure.
The toad variety is Bufo siccus.
Comparative example 1
The selenious acid triglyceride mixture in the step of artificially culturing toad feeding in example 1 is replaced by sodium selenite, and the rest steps are unchanged.
Comparative example 2
The honey in the step of artificially culturing toad feeding in example 1 is removed, and the rest steps are unchanged.
Comparative example 3
The porous microspheres in the step of artificially culturing toad feeding in example 1 are removed, and the rest steps are unchanged.
Comparative example 4
The step of artificially culturing toad feeding in example 1, "S2, pulping: taking out the toad in the S1, cleaning the toad with flowing clear water for 20h, taking out the toad, beating the toad to be stunned, then beating the toad uniformly, adding selenious acid triglyceride mixture and honey, beating the mixture uniformly again to obtain mixed toad pulp, replacing the mixed toad pulp with S2, and pulping: taking out Bufo siccus in S1, washing with flowing clear water for 20 hr, taking out Bufo siccus, beating to make it dizzy, adding selenious acid triglyceride mixture and Mel into Bufo siccus body to obtain mixed Bufo siccus slurry, and keeping the rest steps unchanged.
Comparative example 5
The selenious acid triglyceride mixture in the step of artificially culturing toad feeding in example 1 is removed, and the rest steps are unchanged.
Test example 1
The quality of the toad feeding insects obtained in examples 1-3 and comparative examples 1-5 was recorded, respectively, and the toad feeding yield (toad feeding quality/toad quality) was calculated;
the average selenium content in the toad larvae of examples 1 to 3 and comparative examples 1 to 5 was determined by hydride atomic fluorescence spectrometry according to determination of selenium in national standard food for food safety (GB 5009.93-2017);
randomly selecting 100 toad pests in each of examples 1-3 and comparative examples 1-5, determining the content of the toad peptide in each toad pest according to the method for determining the toad peptide in CN102091318A, and recording the lowest content and the highest content;
100 toad feeding insects in each of examples 1 to 3 and comparative examples 1 to 5 were randomly selected, and the content of the toad active ingredients in the toad feeding insects was expressed by toad venom base content measurement, wherein the measurement method is as follows: quality analysis of toad fed and maggots [ J ] Shenyang pharmaceutical university report, 2013(5), recording minimum and maximum content;
100 toads are randomly selected in each of examples 1 to 3 and comparative examples 1 to 5, the total nitrogen content in the toads is used as the protein content, the total nitrogen content is measured for each toad according to the requirement of the second method of the first appendix IX L of the 2010 edition of the pharmacopoeia of the people's republic of China, and the lowest content and the highest content are recorded.
The test results are shown in Table 1
From table 1, the toad feeding insects obtained in examples 1 to 3 have high yield, high content of bufopeptide and bufogenin, uniform quality, high effective absorption to toads, selenious acid triglyceride is more suitable for enrichment of selenium element by toads, and sodium selenite can also enrich selenium of toads, but has high mortality rate and low final yield; the honey can promote chrysomyia megacephala to soak the mixed toad serous fluid and can help to improve the selenium content and the active ingredients of the toad eating insects; the porous microspheres contain indole, so that the chrysomyia megacephala can be promoted to lay eggs, the egg laying period is shortened, and the yield is improved; compared with the direct eating of toad carcasses, the mixed toad slurry can better ensure the uniform quality of the toad eating insects from the maximum difference content of the toad peptide and the toad venom base.
Test example 2
The toad feeding immune activity research:
experiment one: 120 mice, half male and female, weighing 18-22g, were randomly divided into 4 groups: group I, group II, group III and control group, wherein the toad feeding in example 1 is group I, the toad feeding in comparative example 5 is group II, and the toad feeding in comparative example 5 is group III by adding 2.8% of selenoprotein, pulverizing into fine powder, sieving with 80 mesh sieve, and mixing with 1% of sodium carboxymethyl cellulose to obtain suspension; 0.5mL of 1% sodium carboxymethyl cellulose solution is irrigated to a control group, 0.5mL of 0.332% suspension is irrigated to a group I, a group II and a group III respectively every day, the mice are continuously fed for 15 days, blood is bled from eye sockets of the mice after the last feeding, the animals are sacrificed, the weights of the body weight, the thymus, the liver and the spleen are weighed respectively, and the organ coefficient is calculated;
experiment two: four groups of mice fed in experiment one were bled from the orbit and placed in anticoagulated test tubes with 1% heparin sodium to isolate plasma. Taking 0.1mL of blood plasma to be detected, adding 0.3mL of physiological saline, uniformly mixing, taking 10uL, injecting into a G plate hole, placing in a horizontal wet box, placing in an incubator at 37 ℃, observing the result after 48 hours, reading the diameter of a diffusion ring of each hole by using an immunity measuring instrument, and calculating the content of IgG by using a standard curve;
experiment three: and adding 0.1mL of anticoagulation blood of each group in experiment II into 1.8mL of RPMll640 culture medium, simultaneously adding 0.1mL of PHA into a 37 ℃ water bath for 72h, sucking the supernatant, adding 4mL of human N trampling CL (8.39L /) and uniformly mixing the mixture, carrying out the water bath for 10min, centrifuging the mixture for 10min, discarding the supernatant, adding 5mL of stationary liquid, standing the mixture for 10min at room temperature, and centrifuging the mixture to discard the supernatant. Soaking the glass slide in distilled water, placing the glass slide in a refrigerator, adding 2 drops of precipitate into the glass slide, naturally drying and dyeing, calculating the number of 100 lymphocytes, and calculating the conversion rate;
the results are shown in Table 2:
from table 2, the weight of important immune organs such as thymus and spleen of mice in groups I-III is obviously reduced, immunoglobulin production and lymphocyte transformation rate are also inhibited to a certain extent, which indicates that the toad feeding insects have the same inhibitory action on body humoral immunity and cellular immunity, the mechanism is consistent with that of clinical application of toad feeding insects for treating malignant tumors, compared with the toad feeding insects in comparative example 5 and the mixture of toad feeding insects and selenoprotein in comparative example 5, the toad feeding insects prepared by the method have stronger inhibitory action on body humoral immunity and cellular immunity of mice, and the toad feeding insects prepared by the method have stronger inhibitory activity on malignant tumors.
While the invention has been described in further detail with reference to specific embodiments thereof, it is not intended that the invention be limited to the specific embodiments thereof; for those skilled in the art to which the present invention pertains and related technologies, the extension, operation method and data replacement should fall within the protection scope of the present invention based on the technical solution of the present invention.
Claims (9)
1. The method for artificially culturing toad feeding insects is characterized by comprising the following steps:
s1, cleaning: cleaning living toads with clear water; then putting the toad into the mixed solution for 72 h;
s2, pulping: taking out the toad in S1, washing with flowing clear water for 20-24h, taking out the toad, beating the toad to cause dizziness, then pulping the toad uniformly, adding selenious acid triglyceride mixture and honey, and pulping the mixture uniformly again to obtain mixed toad pulp;
s3, maggot breeding: pouring the mixed toad slurry into a maggot raising pool in a maggot raising box, putting the porous microspheres into a grid box, putting chrysomyia megacephala into a maggot laying chamber, adjusting light, humidity and temperature, starting a ventilation device, starting a magnetic stirring device, continuously eating the mixed toad slurry by the chrysomyia megacephala and performing generation propagation, and obtaining the maximum toad eating number on the fifth day;
s4, maggot collection: turning off the light in the maggot laying chamber, turning on the light in the light chamber, allowing the toad to climb into the maggot collecting chamber through the pore channel, turning on a water sprayer in the maggot collecting chamber, after 2-3h, allowing the toad to climb in, and cleaning to obtain clean fresh maggots;
s5, putting the fresh maggots into water, heating to 48-50 ℃, filtering to remove water after 5-10min, and naturally airing for 2-3 days to obtain toad eating maggots;
the porous microspheres are porous activated carbon soaked in indole; the maggot raising case comprises a maggot raising chamber and a maggot collecting chamber, wherein a maggot raising pool is arranged at the bottom of the maggot raising chamber, a plurality of grid boxes are arranged outside the maggot raising pool, pore passages are arranged outside the grid boxes, and the pore passages are communicated with the maggot raising chamber and the maggot collecting chamber.
2. The method for artificially culturing toad feeding according to claim 1, wherein the porous microspheres are prepared by the following steps: firstly, indole is dissolved in ethanol to prepare the indole with the concentration of 1 × 10-4And (3) pouring the g/mL indole solution into porous activated carbon, immersing, stirring for 30-60min, and drying at 60-70 ℃ for 6-8h to obtain the porous microspheres.
3. The method for artificially culturing toads according to claim 1, wherein the maggot raising case comprises a maggot producing chamber and a maggot collecting chamber, the maggot producing chamber is arranged above the maggot collecting chamber, case shells are arranged outside the maggot producing chamber and the maggot collecting chamber, the case shells are black, an adjustable light, a humidity detector, a spraying device and a ventilation device are arranged at the top of the maggot producing chamber, a maggot raising pool is arranged at the bottom of the maggot producing chamber, magnetic balls are arranged in the maggot raising pool, a magnetic stirrer is arranged below the maggot raising pool, a plurality of grid boxes are arranged outside the maggot raising pool, a pore passage is arranged outside each grid box, the pore passage is communicated with the maggot producing chamber and the maggot collecting chamber, an extension plate is arranged at the lower edge of the pore passage, the extension plate extends outwards, a water sprayer is arranged at the top of the maggot collecting chamber, light chambers are arranged at two sides of the maggot collecting chamber, a baffle plate between the light chamber and the maggot collecting chamber is made of transparent glass, the maggot collecting box is arranged in the middle of the maggot collecting chamber, a cover is not arranged above the collecting box made of transparent glass, a plurality of net holes are uniformly distributed on the maggot collecting box, the net holes do not allow the smallest maggots to pass through, and a water outlet is arranged at the bottom of the maggot collecting chamber.
4. The method for artificially culturing toad feeding according to claim 1, wherein the preparation method of the mixed solution comprises: 100g of magnolia officinalis, 100g of immature bitter orange, 100g of mirabilite and 100g of rhubarb are taken, mixed and crushed, decocted in water for 30min and filtered, 25-30 kg of clean water is added into the filtrate, and the water temperature is kept at 26-28 ℃.
5. The method for artificially cultivating and feeding toad larvae, as claimed in claim 1, wherein the light of both the maggot-producing chamber and the light chamber is variable, ultraviolet light is used for 5-10min before toad slurry is put in, sterilization is carried out, white light is used for 6:00-20:00 after toad slurry is put in, then the light is turned off in 20:00-6:00, the humidity is controlled at 75% ± 5%, and the temperature is controlled at 28 ± 2 ℃; the light of the light chamber is white light when maggots are collected.
6. The method for artificial cultivation of toad feeding insects as claimed in claim 1, wherein the mass quantity ratio of the mixed toad slurry to the Chrysomya megacephala is 1000g/200, wherein the ratio of the Chrysomya megacephala to the Chrysomya megacephala is 1: 1; the female chrysomyia megacephala is to-be-born chrysomyia megacephala.
7. The method for artificially culturing and feeding toads according to claim 1, wherein the chrysomyia megacephala imagoes in the maggot raising box can be continuously used for culturing the toads in the next round after collecting the maggots.
8. The method for artificially culturing and feeding toad pests according to claim 1, wherein the selenious acid triglyceride mixture is synthesized by esterification of sunflower oil and selenious acid, the selenious acid triglyceride content is 2%, and the selenious acid triglyceride mixture mainly has a dioxaselenolane structure.
9. The method for artificially culturing and feeding the toads according to claim 1, wherein the mass-volume ratio of the toads to the selenious acid triglyceride mixture to the honey is as follows: 800-1000g, 20-24g, 50-60 g.
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