Method for simulating rapid induction of anticoccidial drug-resistant strains in field and separation and identification method of anticoccidial drug-resistant strain

A technology of anticoccidiostats and identification methods, which is applied in the direction of microorganism-based methods, separation of microorganisms, biochemical equipment and methods, etc., can solve the problems of time-consuming, complex background of local strains, etc., and achieve short time-consuming, simple operation, Time-sensitive effect

Active Publication Date: 2021-06-04
CHINA AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] For this reason, the technical problem to be solved by the present invention is to provide a method for simulating the rapid induction of coccidiostat-resistant strains in the field, so as to solve the problem of time-consuming and complex background of local strains in the screening of drug-resistant strains in the prior art. In order to quickly and effectively screen a sufficient number of drug-resistant strains with clear background and stable resistance;

Method used

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  • Method for simulating rapid induction of anticoccidial drug-resistant strains in field and separation and identification method of anticoccidial drug-resistant strain
  • Method for simulating rapid induction of anticoccidial drug-resistant strains in field and separation and identification method of anticoccidial drug-resistant strain
  • Method for simulating rapid induction of anticoccidial drug-resistant strains in field and separation and identification method of anticoccidial drug-resistant strain

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] Embodiment 1 simulates the experimental scheme of rapidly inducing salinomycin-resistant strains in the field

[0069] The first batch of induced drug-resistant strain test

[0070] The first batch of experiments set up two groups, that is, the dosing group and the inoculation group. After cleaning and disinfection, they were covered with fresh litter (fresh rice chaff) that had been baked at high temperature and had no coccidia pollution, and equipped with drinking water, food intake, light, Ventilation, insulation and other related equipment. Breeders need to observe the temperature and humidity of the two test groups every day during the test, and improve the test conditions in time according to the data displayed by the temperature and humidity meter, and strictly prevent cross-contamination between groups during the test.

[0071] Introduce 300 1-day-old chicks, and give each chicken a wing number to facilitate the collation of the test results later. And on the ...

Embodiment 2

[0086] Example 2 Drug-resistant strain sample purification and identification

[0087] Oocyst expansion

[0088] Select 7-14 day-old AA broiler chicken coccidia without coccidia infection, orally inoculate chickens with sporulated oocysts, and add a higher dose concentration (3-4 times the working concentration) to the feed 48 hours before inoculation. ) of salinomycin, collect the oocysts in the feces excreted 6-10 days after inoculation.

[0089] Oocyst Purification

[0090] After 48 hours of sporulation, the oocysts were purified. Pour the potassium dichromate containing oocysts into a clean centrifuge tube, centrifuge at 3600rpm for 5min, pour off the upper liquid, resuspend the pellet with PBS, repeat 2-3 times; resuspend the pellet with saturated saline, centrifuge at 3600rpm for 5min , introduce the liquid containing oocysts in the upper layer into another clean centrifuge tube, add 5 times the volume of PBS, and centrifuge at 3600rpm for 5min; pour off the supernata...

Embodiment 3

[0105] Example 3 Simulating the Field Rapid Induction of Drug-resistant Strains Test Program to Obtain Phrysanthone-resistant Insect Strains

[0106] The experiment was divided into two groups. After cleaning and disinfection, they were covered with fresh bedding without coccidia pollution, and equipped with drinking water, feeding, lighting, ventilation, heat preservation and other related equipment. Breeders need to observe the temperature and humidity of the two test groups every day during the test, and improve the test conditions in time according to the data displayed by the temperature and humidity meter, and strictly prevent cross-contamination between groups during the test.

[0107] Introduce 200 1-day-old chicks, and on the day of introducing the coccidia-free chicks, give each chicken a wing number, so that the test records can be made in the later stage. In the dosing group, during the whole test period, a working concentration of the drug (3 mg / kg) was added to t...

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Abstract

The invention belongs to the technical field of coccidium drug-resistant strain induction and acquisition, and particularly relates to a test for rapidly inducing drug-resistant strains in a simulated field and a separation and identification scheme of the drug-resistant strain, which are related to an anticoccidial drug. According to the method for simulating field rapid induction of the anti-coccidiosis drug-resistant strains, by designing two batches of rapid induction drug-resistant strain tests before and after, a plurality of stable drug-resistant strains can be rapidly screened out within 7 weeks, and then a sufficient number of drug-resistant strains can be obtained within a short time. According to the test, under the condition that the original strain background is clear, the coccidium drug-resistant strains can be rapidly screened, high timeliness is achieved, the sample number required by subsequent deep analysis is met, the reliability of the method is verified in the test for screening the drug-resistant strains of the two drugs, and the method has the advantages of being short in consumed time, easy to operate and low in cost and can be widely applied to test induction, screening and identification of chicken coccidiosis drug-resistant strains.

Description

technical field [0001] The invention belongs to the technical field of induction and acquisition of drug-resistant strains of coccidiostats, and in particular relates to a simulated field rapid induction test of drug-resistant strains related to anticoccidiostats and a scheme for the isolation and identification of drug-resistant strains. Background technique [0002] Chicken coccidiosis is a common parasitic disease caused by Eimeria coccidiosis. One of the most costly diseases. Eimeria (Eimeria) belongs to the apicomplexa, and more than 1,800 species have been reported. Seven species of Eimeria have been found and recognized to cause infection, namely, Eimeria tenabilis and Eimeria poisonous. Auris, Eimeria giga, Eimeria heaperia, Eimeria placid, Eimeria precocious, and Eimeria brucei. Coccidia can destroy chicken intestinal epithelial cells. In large-scale farming, coccidiosis outbreaks characterized by high mortality are uncommon. Symptoms such as loss of appetite, lis...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/10C12N1/02C12Q1/686A01K67/02C12R1/90
CPCC12N1/10C12N1/02C12Q1/686A01K67/02C12Q2565/125
Inventor 刘贤勇索勋孙霈郝振凯毕菲菲胡丹丹于咏兰
Owner CHINA AGRI UNIV
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