Case report in Republic of Korea. Dong-Kun Yang Senior researcher Lab. of rabies in viral disease division of QIA

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1 Case report in Republic of Korea Dong-Kun Yang Senior researcher Lab. of rabies in viral disease division of QIA

2 Contents 1. Epidemiological situation of rabies 2. Current control program in wildlife 3. Recent research project related to rabies

3 Case of rabies Epidemiological classification of animal rabies case in Korea Epidemic stage Elimination stage Recurrence stage Year Since the first rabies occurred in 1907, in total, 16,129 rabies cases have been recorded in Korea.

4 No outbreak of animal rabies cases two times up to now Case of rabies ~ 1945: No outbreak (7 years) ~ 1992: No outbreak (8 years) (8 years) Year Korea s policy to control rabies in 1960 s - Implement mass vaccination program to dogs. - Slay stray dogs. - Perform the education for people. As the result, urban rabies was eradicated in Since 1993, sylvatic rabies has been reported in northern border area of Korea.

5 Distribution of rabies cases in Korea from 1993 to DMZ Most of rabies were detected in two provinces near the border of demilitarized zone. Korean raccoon dogs have played a important role in transmission of rabies since 1993.

6 Change of rabies cases in Korea according to provinces Since 2007, rabies cases have not been reported in Gyeonggi province but continued to occur and move to north-eastward in Gangwon province. This result may be obtained from application of bait vaccine to two provinces.

7 Rabies case Biting cases Numbers of human rabies and biting cases in rabies risk regions of Korea Human rabies cases No. of patients bitten by animals Year Year No human rabies occurred between 2005 and Even though incidence of patients bitten by animals in rabies risk regions has been increased, the patients have been treated with postexposure prophylaxis(pep) properly.

8 Case rate(%) Incidence rate of rabies according to animal species Cattle Dog Raccoon dog Others Rabies cases have been diminished in dog. But, since rabies was first confirmed in a Korean raccoon dog in 1994, The cases have been increase.

9 No. of rabies cases Monthly distribution of animal rabies cases from 1993 to 2011 Season (month) Number of cases(%) Spring (March-May) 133 (28.9.) Summer (June-August) 64 (13.9) Autumn (September-November) 97 (21.0) Winter (December-February) 167 (36.2) Total 461 (100) Jan Feb Mar Apr May Jun Jul Aug Sep Oct Nov Dec The monthly distribution of animal rabies during 18 years period peaked in Jan. The incidence rate was the highest during winter. That is because raccoon dogs did not accumulate enough fat in their body for hibernation and came down to villages to seek food.

10 Identification of Korean rabies isolates using IFA test and electronic microscopy The confirmative diagnostic method is fluoresent antibody test(fat). And an asset of virus isolation (VI) is availability for further characterization. Most of rabies isolates are propagated well in neuroblastoma cells (NG ).

11 Application of RT-PCR and rapid kit to rabid brain samples M NC PC M M NC PC M RT-PCR and rapid immunodiagnostic kit can also be used for the detection of rabies in brain or saliva samples of infected animals to make a rapid decision. These kinds of methods could be used as supplementary diagnostic tools. 11

12 Nishigahara SKRDG9902GY Ni-CE SKRDG9901GY RV-97 SKRDG0203CW 100 Gyeonggi SKRBV9801YC PV SKRRD9902PJ NNV-RAB-H 99 ERA Serotype NeiMeng1025C SKRRD9901PJ SAD CVS-11 SKRRD9903YG Flury-LEP 99 SKRDG0204CW F02 94 SKRBV0404HC RRVON D02 SKRRD0406CC 98 Gangwon I SKRBV0403CW 72 FJ009 SKRRD0205HC SKRDG0204HC KRVR0804 KRVB0903 KRVB1001 KRVR KRVB KRVB0907 Gangwon II KRVB KRVB0904 KRVB0902 KRVR0906 KRVR0803 KRVR0901 KRVC0802 KRVB0909 KRVB0908 KRVB1005 KRVB1004 KRVB0910 KRVB1003 KRVB0905 Korean rabies isolates belong to serotype I, genotype I. Based on the phylogenetic tree, Korean rabies isolates classified into 3 subclades. and were most closely related to the eastern Chinese and Russian strains. Korean isolates originated from a rabid animal in northeastern Asia.

13 Multiple alignments of amino acid sequences of glycoprotein from the 14 Korean isolates Isolates KRVR0801 PNNLVVEDEGCTNLSGF...GFGKAYTIFNKTLMEADAHYKSVRTWNEIIPSKGCLR KRVC D KRVR KRVR KRVR KRVB H KRVB KRVB KRVB KRVR KRVB KRVB KRVB KRVB The arginin(r) at position 333 of the residue within the ectodomain of glycoprotein is essential for virulence. There were no changes of arginin among isolates. Therefore, rabies isolates circulating in Korea may pathogenic in several hosts.

14 Rabies case report occurred in March 2011 Species: cattle (Hanwoo), about 5 years old. Date: 21 April Location: Bukmyeon Inje Gangwon province Latitude: 38.06,43 Longitude: Symptom: severe salivation, extreme tremors ataxia, paralysis. Diagnosis: rabies. Indication: All cattle of farm must be vaccinated. According to statement from farmer, wild animal, raccoon dogs were seen wandering Around farm frequently. Like this, recent rabies cases were related to attacks by raccoon dogs. Indications are transferred to farmer associated with rabies

15 Current control program of rabies in Korea

16 Current Korea s policy to control rabies National animal rabies eradication program 1. Annual vaccination of dogs all around of Korea 2. Obligatory vaccination of cattle in rabies risk area 3. Distribution of bait vaccines for wild animals, raccoon dogs 4. Carry out serosurveillance of rabies in two provinces 5. Education for people

17 Amount of distribution(x10,000) Rate of intake(%) Amount of bait vaccine to be distributed into two provinces and intake rate Year Year About 500 thousand doses were distributed in The amount of bait vaccine has been regulated according to the incidence of region. The points distributing bait vaccine are various ranging from 20,734 to 39,280. The time of distribution is spring and autumn twice a year.

18 Density(head/100ha) Korean raccoon dog involved in rabies Family: Canidae, Nycteeutesprocyonoides koreensis American raccoon dog 8 Gangwon Gyeonggi 4% 4% 7 18% 26% 48% Mount Hill Korean raccoon dogs do not bark and live near farm. Population density of raccoon dogs is higher in Ganwon than in Gyeonggi.

19 A process of distributing bait vaccine to field Before distribution of bait vaccine, conduct education (wear gum glove), the vaccines were distributed near farm and small stream. Check locations and the vaccine three times each week. If no trace of taking bait vaccine, change the location of distribution. After 2 month collect them and check the remaining vaccine and report them.

20 Positive rate(%) Seroprevalence of rabies virus in domestic animals of Ganwon province Canine Bovine Year We need continuous surveillance to check national eradication program. From the result, about 70% of dogs demonstrated sero-positive against rabies. It is well known that the rabies is not popular if the antibody ratio of positivity is 70% or greater.

21 Recent research project related to rabies

22 Prepare the ingredients of bait Poultry meal Fish meal Soybean cake Tetracycline Corn Feather meal Sugar Poultry oil + Beta dextrin premix bait

23 Strategy for developing candidate for bait vaccine using reverse genetics system T7 promoter N P M G(mutate) L ERA genome plasmid rerag333glu T7 v v v BHK/T7-9 cell N P L RC-HL helper plasmids The minimal infection unit of negative sense RNA genome is ribonucleoprotein and N, P, L genes are needed to recover recombinant virus

24 Viral titer(10faid50/ml) Result of construction of recombinant rabies virus and propagation in cells M G1 G2 N1 N2 rerag333(bhk) rerag333(ng108) rerag333 strain Days after inoculation Control cells The recombinant vaccine candidate is propagated well in both BHK and NG108 cells. After checking immunogenicity and efficacy of candidate, this strain will be Used for oral bait vaccine seed.

25 New strategy for the point infection control (PIC) program ORV (oral vaccine) TVR PR (population reduction) Location of outbreak Trap-vaccination-release (TVR) program had been performed for 7 years. The Ontario state was declared with rabies free region at October Concentric rings around the location of a rabid animal represent vector population reduction (PR), TVR and ORV zones. It is important to get national model to control rabies.

26 Conclusion National eradication program including massive vaccination and distribution of bait vaccine for the control of rabies in wild animals was lead to a substantial decrease in number of rabies in Korea. When the new reinforced program is prepared and applied to the rabies risk areas, sylvatic rabies will be eradicated in Korea in the near future. These kinds of skills will be transferred to Asian countries.

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