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CN113068767A - Application of sarmentosum bungei extract in preparation of additive for preventing and treating avian aspergillosis - Google Patents

Application of sarmentosum bungei extract in preparation of additive for preventing and treating avian aspergillosis Download PDF

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Publication number
CN113068767A
CN113068767A CN202110341832.XA CN202110341832A CN113068767A CN 113068767 A CN113068767 A CN 113068767A CN 202110341832 A CN202110341832 A CN 202110341832A CN 113068767 A CN113068767 A CN 113068767A
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China
Prior art keywords
extract
sarmentosum
aspergillus
aspergillosis
piper
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Pending
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CN202110341832.XA
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Chinese (zh)
Inventor
段瑞军
戴好富
梅文莉
王昊
盖翠娟
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Institute of Tropical Bioscience and Biotechnology Chinese Academy of Tropical Agricultural Sciences
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Institute of Tropical Bioscience and Biotechnology Chinese Academy of Tropical Agricultural Sciences
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Priority to CN202110341832.XA priority Critical patent/CN113068767A/en
Priority to PCT/CN2021/092030 priority patent/WO2022205557A1/en
Publication of CN113068767A publication Critical patent/CN113068767A/en
Pending legal-status Critical Current

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    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23KFODDER
    • A23K10/00Animal feeding-stuffs
    • A23K10/30Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23KFODDER
    • A23K50/00Feeding-stuffs specially adapted for particular animals
    • A23K50/70Feeding-stuffs specially adapted for particular animals for birds
    • A23K50/75Feeding-stuffs specially adapted for particular animals for birds for poultry
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18Magnoliophyta (angiosperms)
    • A61K36/185Magnoliopsida (dicotyledons)
    • A61K36/67Piperaceae (Pepper family), e.g. Jamaican pepper or kava
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/10Antimycotics
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/30Extraction of the material
    • A61K2236/33Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
    • A61K2236/331Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation or decoction
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/30Extraction of the material
    • A61K2236/33Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
    • A61K2236/333Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/30Extraction of the material
    • A61K2236/39Complex extraction schemes, e.g. fractionation or repeated extraction steps
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/50Methods involving additional extraction steps
    • A61K2236/51Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Natural Medicines & Medicinal Plants (AREA)
  • Polymers & Plastics (AREA)
  • Botany (AREA)
  • Animal Husbandry (AREA)
  • Biotechnology (AREA)
  • Medicinal Chemistry (AREA)
  • Food Science & Technology (AREA)
  • Mycology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Birds (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Zoology (AREA)
  • Medical Informatics (AREA)
  • Physiology (AREA)
  • Molecular Biology (AREA)
  • Epidemiology (AREA)
  • Microbiology (AREA)
  • Communicable Diseases (AREA)
  • Oncology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Organic Chemistry (AREA)
  • Alternative & Traditional Medicine (AREA)
  • Medicines Containing Plant Substances (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

The invention relates to the technical field of poultry breeding, in particular to application of sarmentosum bungei extract in preparing an additive for preventing and treating poultry aspergillosis. The research of the invention finds that the laetispicum aqueous extract and the alcohol extract have an inhibiting effect on aspergillus funguses such as aspergillus fumigatus, aspergillus flavus, aspergillus niger, aspergillus terreus and the like, and verifies that the laetispicum extract can be used for preventing and treating respiratory tract and intestinal tract diseases of poultry caused by aspergillus infection, and the effective rate can reach 81-84%. In addition, the sarmentosum bungei has the advantages of wide plant resource distribution, large biomass and low price, is particularly visible under the Hainan forest and everywhere on the roadside, and the extract vegetation has the advantages of simple process, low cost, high yield and easy large-scale production.

Description

Application of sarmentosum bungei extract in preparation of additive for preventing and treating avian aspergillosis
Technical Field
The invention relates to the technical field of poultry breeding, in particular to application of sarmentosum bungei extract in preparing an additive for preventing and treating poultry aspergillosis.
Background
The avian aspergillosis is a common fungal disease in the duck breeding industry, and particularly, the avian aspergillosis has more morbidity in young ducks and higher mortality rate which can reach 50%. The pathogenic bacteria causing avian aspergillosis are aspergillus including aspergillus fumigatus, aspergillus flavus, aspergillus niger, aspergillus terreus, aspergillus niger and the like. In Hainan weather, the temperature is high and the humidity is high, feed and bedding are easy to mildew, and more mould spores exist in the culture environment, and the disease is a common disease in duck culture in the summer of Hainan. The clinical symptoms of the disease are dyspnea, gasp by mouth opening, increased breathing frequency, inappetence, preference for drinking water, love for sleeping, light green and thin stool, frequent staying or lying in the corner, and viscous secretion flowing out of the oral cavity and the nasal cavity. At present, no specific medicine is available for treating the disease, and the treatment effect is obvious when 1 thousandth of copper sulfate, 1 thousandth of potassium iodide, 0.05 percent of clotrimazole and 0.5 ten thousand IU of nystatin are used.
Along with the development of economy in China, the living standard of people is increasingly improved, and the requirements on the quality of meat products are also continuously improved. The Chinese herbal medicine is one of the development trends to replace antibiotics to be used as a feed additive, has important significance for green and healthy development of duck breeding industry, and plays an important role in providing healthy meat and eggs for society. However, no report about Chinese herbal medicines for preventing and treating duck aspergillosis is found at present.
Disclosure of Invention
In view of the above, the technical problem to be solved by the present invention is to provide an application of sarmentosum bungei extract in preparation of feed additives.
The invention provides application of sarmentosum bungei extract in preparation of an additive for preventing and treating avian aspergillosis.
In the invention, the poultry aspergillosis is caused by aspergillus fumigatus, aspergillus flavus, aspergillus niger, aspergillus terreus and aspergillus niger.
In the invention, the poultry aspergillosis is duck aspergillosis.
In some embodiments, the duck aspergillosis is aspergillosis of a red-haired duck.
In the invention, the sarmentosum bungei extract is an aqueous extract or an alcohol extract of sarmentosum bungei.
In the embodiment of the invention, the preparation method of the laetispicum water extract comprises the steps of extracting laetispicum for three times by refluxing with water, combining the extracting solutions, and concentrating to obtain the laetispicum water extract; in the three extractions, the first extraction lasts for 3 hours, the second extraction lasts for 2 hours, and the third extraction lasts for 1 hour. In the first extraction, the mass volume ratio of the piper sarmentosum to the water is 1 kg: 0.5L; in the second extraction, the mass volume ratio of the piper sarmentosum to the water is 1 kg: 0.4L; in the third extraction, the mass volume ratio of caulis et folium Piperis Hancei to water is 1 kg: 0.3L.
In the embodiment of the invention, the preparation method of the ethanol extract of the sarmentosum bungei comprises the steps of leaching the sarmentosum bungei with ethanol for three times, combining the extracting solutions, and concentrating to obtain the ethanol extract of the sarmentosum bungei; in the three extractions, the first extraction is carried out for 7 days, the second extraction is carried out for 5 days, and the third extraction is carried out for 3 days. In the first extraction, the mass volume ratio of the piper sarmentosum to the ethanol is 1 kg: 2L; in the second extraction, the mass volume ratio of the sarmentosum bungei to the ethanol is 1 kg: 1.5L; in the third extraction, the mass volume ratio of the sarmentosum bungei to the ethanol is 1kg to 1.5L. The ethanol is 95% ethanol. I.e. a solution of ethanol-water mixed in a volume ratio of 95: 5.
The invention also provides an additive comprising sarmentosum bungei extract.
The additive of the present invention may be added to feed or drinking water, but the present invention is not limited thereto. The addition amount of the water-soluble organic fertilizer in drinking water is 1%. Namely, the weight percentage of the sarmentosum bunge extract added into drinking water is 1 percent.
The invention also provides a method for raising poultry, which is to administer the additive.
In the present example, the administration time was 7 days old, and the administration was continued for one week.
The research of the invention finds that the laetispicum aqueous extract and the alcohol extract have an inhibiting effect on aspergillus funguses such as aspergillus fumigatus, aspergillus flavus, aspergillus niger, aspergillus terreus and the like, and verifies that the laetispicum extract can be used for preventing and treating respiratory tract and intestinal tract diseases of poultry caused by aspergillus infection, and the effective rate can reach 81-84%. In addition, the sarmentosum bungei has the advantages of wide plant resource distribution, large biomass, low price, simple preparation process, low cost, high yield and easy large-scale production, and is particularly visible under Hainan forests and roadside places.
Detailed Description
The invention provides the application of the sarmentosum bungei extract in preparing feed additives, and the technical personnel can use the content to reference and appropriately improve the process parameters. It is expressly intended that all such similar substitutes and modifications which would be obvious to one skilled in the art are deemed to be included in the invention. While the methods and applications of this invention have been described in terms of preferred embodiments, it will be apparent to those of ordinary skill in the art that variations and modifications in the methods and applications described herein, as well as other suitable variations and combinations, may be made to implement and use the techniques of this invention without departing from the spirit and scope of the invention.
The test materials adopted by the invention are all common commercial products and can be purchased in the market. The Chinese herbal medicine sarmentosum, namely the Piper betle, the Piper betle Piper betel, the Piper betle, the Piper sarmentosum, the Piper betle, the Piper betel and the Piper betel are Piperaceae (Piperaceae) Piper L.) Piper betel which has the effects of dispelling wind and cold, promoting qi circulation and relieving pain, activating collaterals and relieving swelling and is used for wind-cold cough and asthma, rheumatalgia, abdominal distension, diarrhea, dysentery, postpartum edema and traumatic injury (Chinese materia Medica). The caulis et folium Piperis Hancei extract can be prepared from whole plant of caulis et folium Piperis Hancei, or part of tissue and organ. Before extraction, drying and crushing. In the embodiment of the invention, the sarmentosum bungei extract is prepared by adopting sarmentosum bungei whole grass.
The invention is further illustrated by the following examples:
example 1
Pulverizing dried caulis et folium Piperis Hancei 1kg, placing into 5L triangular flask, adding 95% ethanol 2L, soaking at room temperature for 7 days, and recovering extractive solution; adding 1.5L 95% ethanol, soaking for 5 days, and recovering extractive solution; adding 1.5L 95% ethanol, soaking for 3 days, recovering extractive solution, mixing 3 times of extractive solutions, and concentrating under reduced pressure to obtain 110g ethanol extract of Piper hancei.
Example 2
Pulverizing dried caulis et folium Piperis Hancei 1kg, placing into 2L round bottom flask, adding water 0.5L, connecting with condensing system, heating and boiling for 3 hr, and recovering extractive solution; adding 0.4L water, heating and boiling for 2 hr, and recovering extractive solution; adding 0.3L water again, heating and boiling for 1 hr, recovering extractive solution, mixing 3 times of leaching solutions, and concentrating under reduced pressure to obtain 150g of caulis et folium Piperis Hancei water extract.
Example 3
Bacteriostatic experiments on the sarmentosum bungei extracts prepared in examples 1 and 2
Activating target strain, and diluting to 1 × 106CFU/mL bacterial liquid is reserved; preparing a YPD sterile plate; uniformly coating 0.1mL of prepared bacterial liquid on an YPD flat plate by using a sterile cotton swab; dissolving the sarmentosum bungei extract obtained in the embodiment 1-2 in methanol respectively to prepare a solution with the concentration of 10 mg/mL; preparing 10mg/mL solution of clotrimazole with acetone, dripping 10 μ L of the solution onto 6 mm-diameter sterile filter paper respectively, placing the filter paper on plates coated with target bacteria solution after the solvent is volatilized, placing 6 pieces of filter paper on each plate, sealing the plates, placing the plates upside down in an incubator at 28 ℃ for culturing for 24 hours, and measuring the diameter (mm) of a bacteriostatic spot with a vernier caliper. The target bacteria used in the bacteriostasis experiment are aspergillus fumigatus, aspergillus niger and aspergillus flavus; 3 replicates of each bacteria inhibition experiment were set up, each replicate comprising 2 pieces of filter paper sheets each containing 10mg/mL of sarmentosum extract, 10mg/mL of clotrimazole (positive control), methanol (blank control). The test results are shown in table 1:
TABLE 1 bacteriostasis test (unit mm) of the Piper sarmentosum extract obtained in examples 1 to 2
Aspergillus fumigatus Aspergillus niger Aspergillus flavus
Example 1 extract 21.41±1.71 27.35±1.62 18.14±1.22
Example 2 extract 22.15±1.31 26.72±1.47 20.36±1.43
Clotrimazole 24.36±1.17 28.43±1.75 21.78±1.64
Methanol 0±0 0±0 0±0
As can be seen from in vitro bacteriostasis experiments of data in Table 1, the sarmentosum bungei extracts prepared by the two methods have better inhibiting effect on 3 bacteria in Aspergillus, and the 3 aspergillosis are the most common pathogenic bacteria causing duck aspergillosis. The experiments show that the sarmentosum bungei extract can be used for preventing and controlling the duck aspergillosis.
Example 4
Bacteriostatic experiments on the sarmentosum bungei extracts prepared in examples 1 and 2 on the basis of example 3
The sarmentosum bungei extract was dissolved in DMSO to an initial concentration of 1mg/mL, clotrimazole was used as a positive control and DMSO was used as a blank control to dissolve in DMSO to an initial concentration of 1 mg/mL. And (3) performing gradient dilution on a 96-well plate by using a sterile culture medium according to a 2-fold method, adding bacteria to be detected after dilution, and observing after culturing for 12 hours. This experiment was set up in 3 replicates.
TABLE 2 MIC test (unit. mu.g/mL) of sarmentosum extract obtained in examples 1 to 2
Aspergillus fumigatus Aspergillus niger Aspergillus flavus
Example 1 extract 10~25 25~50 50~75
Example 2 extract 25~50 10~25 25~50
Clotrimazole 0.65 0.65 1.26
DMSO >100 >100 >100
From MIC experimental data in Table 2, the minimum inhibitory concentration of the piper sarmentosum extract on 3 bacteria in Aspergillus is less than 100 mu g/mL, which shows that the extract has a certain inhibitory effect on the above 3 Aspergillus and can be used as a feed additive for preventing and controlling duck aspergillosis.
Example 5
Examples 1 and 2 Duck aspergillosis control test of sarmentosum extract
The test is carried out in a duck farm in a certain paddy field in the state of delirium, the test time is 7 months, the test is the 7-day-old seedling of the red-feather duck in the seasons of high temperature and high humidity in Hainan and high occurrence of aspergillosis. 120 7-day-old ducks with confirmed aspergillosis were selected for the test in 4 groups of 30 ducks. The test subjects were fed continuously for 1 week with a test using a 1% aqueous solution of sarmentosum extract, a 0.05% aqueous solution of clotrimazole and water (blank control) instead of drinking water, and the treatment effect was evaluated according to the following criteria:
and (3) curing: the duck appetite and activity spirit are recovered, the color of the excrement is normal, and the dryness and the thinness are moderate.
Death: the disease of the ducks is not obviously improved and the ducks die in the prevention and treatment period.
The method has the following advantages: the appetite and activity spirit of the ducks are obviously improved, the diarrhea frequency is reduced, and the effective number comprises the cure number.
And (4) invalidation: the disease condition of the ducks is not obviously improved or even aggravated during the prevention and treatment period, but the death number is not included. The evaluation results are shown in Table 3.
TABLE 3 Duck aspergillosis control test of sarmentosum extract prepared in examples 1-2
The mortality rate is% The cure rate is% Effective rate% Not efficiency%
Example 1 extract 6.67 73.33 83.33 10.00
Example 2 extract 6.67 70.00 80.00 13.33
Clotrimazole 3.33 83.33 90.00 6.67
Drinking water 53.33 6.67 13.33 33.33
As can be seen from the data in Table 3, the piper sarmentosum extract has a good effect of preventing and controlling duck mycosis caused by aspergillus, and the effect is slightly poor compared with the clotrimazole effect of a control group, but the piper sarmentosum extract can be used as a feed additive for preventing and controlling duck mycosis.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that it is obvious to those skilled in the art that various modifications and improvements can be made without departing from the principle of the present invention, and these modifications and improvements should also be considered as the protection scope of the present invention.

Claims (10)

1. Application of caulis et folium piperis sarmentosi extract in preparing additive for preventing and treating avian aspergillosis is provided.
2. Use according to claim 1, characterized in that the avian aspergillosis is caused by aspergillus fumigatus, aspergillus flavus, aspergillus niger, aspergillus terreus and aspergillus niger.
3. Use according to claim 1, wherein the avian aspergillosis is duck aspergillosis.
4. The use according to claim 3, wherein the duck aspergillosis is aspergillosis of a red-haired duck.
5. The use according to any one of claims 1 to 4, wherein the Piper hancei extract is an aqueous extract or an alcoholic extract of Piper hancei.
6. The use of claim 5, wherein the aqueous extract of Piper sarmentosum is prepared by extracting Piper sarmentosum with water under reflux three times, combining the extracts and concentrating to obtain an aqueous extract of Piper sarmentosum; in the three extractions, the first extraction lasts for 3 hours, the second extraction lasts for 2 hours, and the third extraction lasts for 1 hour.
7. The use of claim 5, wherein the alcohol extract of Piper sarmentosum is prepared by extracting Piper sarmentosum with ethanol three times, combining the extracts, and concentrating to obtain alcohol extract of Piper sarmentosum; in the three extractions, the first extraction is carried out for 7 days, the second extraction is carried out for 5 days, and the third extraction is carried out for 3 days.
8. An additive, comprising sarmentosum bungei extract.
9. A method for feeding birds comprising administering the supplement of claim 8.
10. The method of claim 9, wherein the administration is for a period of 7 days of age and is for one week.
CN202110341832.XA 2021-03-30 2021-03-30 Application of sarmentosum bungei extract in preparation of additive for preventing and treating avian aspergillosis Pending CN113068767A (en)

Priority Applications (2)

Application Number Priority Date Filing Date Title
CN202110341832.XA CN113068767A (en) 2021-03-30 2021-03-30 Application of sarmentosum bungei extract in preparation of additive for preventing and treating avian aspergillosis
PCT/CN2021/092030 WO2022205557A1 (en) 2021-03-30 2021-05-07 Use of piper sarmentosum roxb extract in preparation of additive for preventing and treating avian aspergillosis

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CN202110341832.XA CN113068767A (en) 2021-03-30 2021-03-30 Application of sarmentosum bungei extract in preparation of additive for preventing and treating avian aspergillosis

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Application publication date: 20210706