A  Preliminary Study of “Minzhi peculiar smell eliminator”

to Prevent from Infection Avian influenza Virus

敏芝异味消防治禽流感试验报告

QingJing Smell Science Research Institute，Xiangcheng district, Xiangfan City, Hubei Province ,China, 

Li De-Min 李德敏

 Abstract: In order to study the effect of antiseptic “Minzhi peculiar smell eliminator”(designated as peculiar smell eliminator), a botanical based disinfectants were tested for the prevention chickens from being infected by H5N1 highly pathogenic avian influenza virus. A protection trail that SPF chickens administrated medicines in advance by routes of spray, drink water and feedstuffs, lived with chickens infected with virus, was carried out. The result: “peculiar smell eliminator” diluted in 1:10 with saline, is non-toxic to chickens embryos 66.7% avian influenza viruses were inactivated in 15 minutes by “peculiar smell eliminator” diluted in 1:5. “peculiar smell eliminator” was sprayed chickens directly, mixed in drinking water and feedstuff for 3 days prior to experiment. Then the chickens could be kept from being infected by H5N1 virus. The rate of prevention and protection was 100%.

Materials and methods

1. Medicine: “peculiar smell eliminator”, provided by Qingjing Smell Science Research Institute，Xiangcheng District, Xiangfan City,

2. Virus: Avian influenza virus A/Goose/Guangdong/1/96 (H5N1) designated as GD/1/96, 2×10⁹ EID₅₀/ml(50% egg infectious dose).

3. Specific pathogen free (SPF) chicken embryos and SPF chickens: 9~11 days old chicken embryos and 6-week-old chickens, provided by the experimental animal center of Harbin Veterinary Research Institute of Chinese Academy of Agriculture Science, SPF chicken were raised in minus pressure segregated incubator.

4. Chicken embryo toxicity test: “peculiar smell eliminator” was diluted with PBS solution, injected into SPF chicken embryoed eggs, the chicken embryo survival rate was checked up after incubation for 48 hours at 37℃.

5. Efficacy test for influenza virus in vivo: Disinfectants were diluted and mixed with H5N1 avian influenza virus at different ratio, the disinfecting efficacy was checked by injection of the mixture of virus and disinfectants into chicken embryoed eggs, and the virus left over was tested by chicken red cells hemagglutination test.

6. Grouping of chickens: sixty, six-week-old SPF chickens were randomly divided into three groups as the followings: test group, control group and virus infected group, respectively.

7. Administration methods of the disinfectants “peculiar smell eliminator”: chickens of test group were administrated the disinfectants as the following ways: the first day drinking water contained 1%“peculiar smell eliminator”, the second day drinking water contained 2%“peculiar smell eliminator”, the third day drinking water contained 3%“peculiar smell eliminator” since the fourth day, drinking water contained 4%“peculiar smell eliminator” daily throughout the whole experiment, when drunk water contain drug, fed with feed mixed with drug and pasteurized spreading in segregated machinery air, 3 times everyday, once every 4 hours, were carried out.

8. Methods of virus infection: 20 SPF chickens in infected group were inoculated with avian influenza virus GD/1/96 through dropping nose and sprinkling eyes, which inoculation dose was 10⁶EID₅₀ every chicken. After infected virus 3hours, 10 infected chickens were put in segregated machinery of trail group and control group, 5every group, lived and fed mixture with other chickens. clinical feature every chickens groups were observed and recorded everyday.

9. Isolation of Virus: On day 2 and day 5 after the chicken infected with virus, samples were taken by trachea and cloacae swabbing, and then inoculating the SPF chicken embryos respectively to isolate the virus, the sub-type of virus were identified by hemagglutination test and hemagglutination inhibition test.

Results

1. Toxicity Test

The result indicated that “peculiar smell eliminator” is not toxic to chicken embryos inoculated in concentration of 1:10 in water.

2. Avian influenza virus destroyed in vitro by “peculiar smell eliminator”:

All virus were killed in 15 minutes under room temperature when the virus were mixed with “peculiar smell eliminator”66.7 percent of the virus were killed in 15 minutes when “peculiar smell eliminator” and virus were mixed in ratio of 1:5 30 percent of the virus were killed in 30 minutes when “peculiar smell eliminator” and virus were mixed in ratio of 1:10. The result of test as below:

Note : the dosage of the H5N1 avian influenza virus is 10⁶EID₅₀/0.2ml the embryos were inoculated with avian influenza virus that was mixed with “peculiar smell eliminator” in different times under room temperature.

3. Result of clinic trials with “peculiar smell eliminator”: “peculiar smell eliminator” administrated by spray, mixed in drinking water or feed is 100 percent (20/20)effective preventing tested chickens from infection by virus-infected chickens. The virus isolation results show all of the “peculiar smell eliminator” treated chickens are virus negative while the chickens of the control group and infected group are virus positive. All the chickens of the infected group died in five day post virus infected by nasal and eye route directly. The chickens of control group lived with the infected chicken together are all dead by ninth days, and viruses are isolated from these chickens.

Conclusions

peculiar smell eliminator, the disinfects made from botanic materials, is not of toxic for chicken embryo inoculated in concentration of 1:10 dilution 66.7% of avian influenza virus is inactivated by“peculiar smell eliminator” at the concentration of 1:5 dilution in 15 minutes.    peculiar smell eliminator administrated to chickens by three routes: sprayed directly, mixed in drinking water and mixed in feed, are 100 percent(20/20) effective preventing H5N1 avian influenza virus from infection.

 This Drug test reports has no references.