Infections of Two Isthmiophora Species (Digenea: Echinostomatidae) in Wild Mammals from Republic of Korea with Their Morphological Descriptions

Isthmiophora hortensis (Digenea: Echinostomatidae) is a dominant echinostome in animal reservoir hosts and humans in the Republic of Korea (Korea). We intended to investigate the infection status with this echinostome species in the several species of wild animals and describe the morphological characteristics in the faunistic view point. A total of 175 animal carcasses belonging to 3 families, i.e., Canidae, Felidae and Mustelidae, were collected from the southern regions of Korea from March 2010 to July 2017. Isthmiophora spp. worms were recovered from the small intestines of each animal under a stereomicroscope after washing of intestinal contents. Isthmiophora hortensis was recovered from 4 species of wild carnivores, i.e., Nyctereutes procyonoides (3/107: 2.8%), Mustela sibirica (11/31: 35.5%), Meles lucurus (2/3: 33.3%) and Martes flavigula (1/2: 50%). The other 3 carnivores comprising stray dogs, cat and leopard cat were negative for I. hortensis infection (0/2, 0/10 and 0/12, respectively). Specimens obtained from the Lutra lutra (6/8: 75%) were identified as a distinct species, I. inermis, by morphological comparison. Isthmiophora inermis has thinner body, elongate testes and different anterior limits of vitelline fields. Detailed morphological descriptions and comparisons with the morphological characteristics are provided. Conclusively, it was confirmed for the first time that 3 species of mustelid mammals, i.e., M. sibirica, M. lucurus, and M. flavigula, are to be the new definitive hosts of I. hortensis in Korea. Additionally, I. inermis is to be newly added in the Korean echinostome fauna.

Rabies immune status of raccoon dogs residing in areas where rabies bait vaccine has been distributed

Since 2000, large amounts of rabies bait vaccine have been distributed in two provinces where raccoon dog-mediated rabies has occurred. A total of 146 raccoon dogs were caught in Gangwon and Gyeonggi Provinces from January 2017 to June 2018, and raccoon dog blood samples were collected. Of the 146 raccoon dogs, 13.7% (20/146) had rabies antibodies. In Gyeonggi and Gangwon provinces, the rate of rabies antibody was 8.5% (5/59) and 17.2% (15/87), respectively. Considering these results, it would be desirable to improve the distribution method or use a new bait vaccine to prevent animal rabies in South Korea.

Morphological and Molecular Characterization of Toxocara tanuki (Nematoda: Ascaridae) from Korean Raccoon Dog, Nyctereutes procyonoides koreensis

Present study was performed to describe the morphological and molecular characterization of Toxocara tanuki (Nematoda: Ascaridae) from Korean raccoon dog, Nyctereutes procyonoides koreensis, naturally infected in the Republic of Korea (Korea). Juvenile and adult worms of T. tanuki were recovered in 5 out of 10 raccoon dogs examined and the larval worms were detected in 15 out of 20 muscle samples (75%). Small lateral alae were observed on the cranial end of the body in male and female adults and 2 long spicules (3.0–3.5 mm) were characteristically observed in the posterior end of males. In SEM observation, 18 pairs of proximal precloacal, a precloacal median, a postcloacal median and 5 pairs of postcloacal papillae were uniquely revealed in the posterior portion of males, but the proximal papillae were not shown in the lateral ends of females. Molecular analysis on the 18S rRNA partial DNA sequences was revealed the same finding in both samples, adult worms and muscle larvae, which are closely related to T. tanuki. In conclusion, it was confirmed for the first time that T. tanuki is indigenously distributed, the Korean raccoon dog is acted as the natural definitive host of this nematode in Korea and the morphological characteristics of T. tanuki were shown in specific structure for single postcloacal median papilla in male.

Incidence and Sero-survey of Canine Adenovirus Type 2 in Various Animal Species

Canine adenovirus type 2 (CAV-2) is the cause of a major respiratory illness in dogs. In this study, we analyzed adenovirus infections in dogs using 2000–2017 data from the Animal and Plant Quarantine Agency (APQA) and conducted a serological survey of CAV-2 infection in six animal species in Korea. In total, 38 of the 3,179 dog samples were confirmed as canine adenovirus infections. In serological survey, 1,028 dog sera, 160 raccoon dog sera, 100 cattle sera, 257 sow sera, 206 horse sera, and 106 cat sera, collected from January 2016 to July 2018, were screened for the presence of anti-CAV-2 antibodies by virus neutralization test. The seropositivity rates for dogs, raccoon dogs, cattle, sows, horses, and cats were 88.5% (910/1,028), 51.3% (82/160), 85.0% (85/100), 48.6% (125/257), 35.0% (72/206), and 2.8% (3/106), respectively. Among dogs and raccoon dogs, 1.9% (20/1,028) and 8.8% (14/160), respectively, had a virus-neutralizing antibody (VNA) titer of over 1:256. A high CAV-2 VNA titer indicates a repeated vaccination or natural infection in Korean dogs and circulation of CAV-2 in raccoon dog populations.

Serological and Molecular Detection of Toxoplasma gondii and Babesia microti in the Blood of Rescued Wild Animals in Gangwon-do (Province), Korea

Infections of Toxoplasma gondii and Babesia microti are reported in many wild animals worldwide, but information on their incidence and molecular detection in Korean wild fields is limited. In this study, the prevalence of T. gondii and B. microti infection in blood samples of 5 animal species (37 Chinese water deer, 23 raccoon dogs, 6 roe deer, 1 wild boar, and 3 Eurasian badgers) was examined during 2008–2009 in Gangwon-do (Province), the Republic of Korea (=Korea) by using serological and molecular tests. The overall seropositivity of T. gondii was 8.6% (6/70); 10.8% in Chinese water deer, 4.3% in raccoon dogs, and 16.7% in roe deer. PCR revealed only 1 case of T. gondii infection in Chinese water deer, and phylogenic analysis showed that the positive isolate was practically identical to the highly pathogenetic strain type I. In B. microti PCR, the positive rate was 5.7% (4/70), including 2 Chinese water deer and 2 Eurasian badgers. Phylogenetic analysis results of 18S rRNA and the β-tubulin gene showed that all positive isolates were US-type B. microti. To our knowledge, this is the first report of B. microti detected in Chinese water deer and Eurasian badger from Korea. These results indicate a potentially high prevalence of T. gondii and B. microti in wild animals of Gangwon-do, Korea. Furthermore, Chinese water deer might act as a reservoir for parasite infections of domestic animals.

Mass vaccination has led to the elimination of rabies since 2014 in South Korea

PURPOSE: Rabies is one of the most fatal diseases, but it is 100% preventable in animals by vaccination. In this study, we present the epidemiological features of, and national preventive measures against, rabies in Korea. MATERIALS AND METHODS: Data related to rabies and the population density of raccoon dogs in Korea were collected from the Animal and Plant Quarantine Agency, the Korean Centers for Disease Control and Prevention, and the National Institute of Environmental Research. Rabies diagnosis was confirmed with a fluorescent antibody test using brain samples of animals in accordance with the procedures described by the World Organization for Animal Health. Serological assays for dogs and cattle were conducted using the fluorescent antibody virus neutralization test. RESULTS: From 1993 to 2016, a total of seven human rabies cases and 437 animal rabies cases in five different species were reported. An increase in the distribution of bait vaccine seemed to be related to a dramatic decrease in rabies prevalence in endemic rabies regions. Two Korean provinces and the capital city, Seoul, were involved in rabies outbreaks. Korean rabies strains are most closely related to the eastern Chinese strain belonging to the Arctic-like lineage. The yearly seropositive rates ranged from 50.4% to 81.2% in dogs and from 25% to 60.5% in cattle residing in endemic rabies regions. CONCLUSION: This study indicates that national preventive measures, including mass vaccination and distribution of bait vaccines, have contributed to a substantial decrease in the number of rabies cases in Korea.

Detection of viral infections in wild Korean raccoon dogs (Nyctereutes procyonoides koreensis)

Wild raccoon dogs (Nyctereutes procyonoides koreensis) may play a role transmitting several pathogens to humans and pet animals. Information concerning the incidence of rabies, canine distemper virus (CDV), canine parvovirus (CPV), canine adenovirus type 2 (CAdV-2), canine parainfluenza virus type 5 (CPIV-5), and canine herpesvirus (CHV) is needed in wild raccoon dogs. In total, 62 brain samples of raccoon dogs were examined for rabies virus (RABV) and CDV, and 49 lung samples were screened for CDV, CAdV-2, CPIV-5, and CHV. No RABV, CAdV-2, CPIV-5, or CHV was identified, but nine CDV antigens (8.1%, 9/111) were detected. Moreover, 174 serum samples from wild raccoon dogs were screened for antibodies against the five major viral pathogens. The overall serosurveillance against CDV, CPV, CAdV-2, CPIV-5, and CHV in wild raccoon dogs was 60.3%, 52.9%, 59.8%, 23.6%, and 10.3%, respectively. Comparisons of the sero-surveillance of the five pathogens showed that raccoon dogs of Gyeonggi province have slightly higher sero-positive rates against CDV, CPV, and CHV than those of Gangwon province. These results indicate high incidences of CDV, CPV, and CAdV-2 in wild raccoon dogs of two Korean provinces and a latent risk of pathogen transmission to companion and domestic animals.

New Definitive Hosts and Differential Body Indices of Isthmiophora hortensis (Digenea: Echinostomatidae)

The present study was performed to record new definitive hosts of Isthmiophora hortensis, and to describe morphological characteristics derived from a variety of worm samples for clarification of its taxonomic validity. Morphological characteristics with dimensions were observed in worm samples (n=21) from naturally infected wild animals, including a raccoon dog Nyctereutes procyonoides from Gimhae-si (City), Gyeongsangnam-do, stray cats and a striped field mouse from several localities, and a wild boar Sus scrofa, from Gurye-gun (County), Jeollanam-do. In addition, adult flukes (n=45) recovered in albino rats experimentally infected with the metacercariae from a freshwater fish species were also subjected to morphological studies. The mean ratios of the body length (BL) to body width (BW) were 5.86 and 5.76 in worms from wild animals and experimental rats, respectively. Those of the ventral sucker to oral sucker were 2.92 and 3.01 in worms from 2 groups. The mean percentages of the hindbody length (HBL) to BL were 42.1 and 41.2 in 2 groups. Those of uterine fields to BL were 9.8 and 12.2 in the 2 worm groups. By the present study, the 2 species of wild animals, the raccoon dog and wild boar, have been added as new definitive hosts for I. hortensis. The morphological characteristics of adult flukes derived from a variety of host source were redescribed to support the taxonomic validity of this echinostome species.

Safety and immunogenicity of recombinant rabies virus (ERAGS) in mice and raccoon dogs

PURPOSE: The development of a genetically modified live rabies vaccine applicable to wild raccoon dogs is necessary for the eradication of rabies in Korea. Thus, we constructed a recombinant rabies virus (RABV) called the ERAGS strain, using a reverse genetic system and evaluated its safety and efficacy in mice and its safety and immunogenicity in raccoon dogs. MATERIALS AND METHODS: ERAGS, which has Asn194Ser and Arg333Glu substitutions in the glycoprotein, was constructed using site-directed mutagenesis. Mice were inoculated with the ERAGS strain (either 10(5.0) or 10(7.0) FAID(50)/mL) via intramuscular (IM) or intracranial injections and then challenged with a virulent RABV. Raccoon dogs were administered the ERAGS strain (10(8.0) FAID(50)/mL) either orally or via the IM route and the immunogenicity of the strain was evaluated using fluorescent antibody virus neutralization tests. RESULTS: The ERAGS strain inoculated into murine neuroblastoma cells reached 10(7.8) FAID(50)/mL at 96-hour post-inoculation. The virus was not pathogenic and induced complete protection from virulent RABV in immunized 4- and 6-week-old mice. Korean raccoon dogs immunized with the ERAGS strain via IM or oral route were also safe from the virus and developed high titer levels (26.4-32.8 IU/mL) of virus-neutralizing antibody (VNA) at 4 weeks post-inoculation. CONCLUSION: The ERAGS RABV strain was effectively protective against rabies in mice and produced a high VNA titer in raccoon dogs.

Oral immunization of mice with recombinant rabies vaccine strain (ERAG3G) induces complete protection

PURPOSE: New rabies vaccine bait for both pets and raccoon dogs residing in Korea is needed to eradicate rabies infection among animals. In this study, we constructed a recombinant rabies virus (RABV), the ERAG3G strain, using a reverse genetics system. Then we investigated the efficacy of this strain in mice after oral administration and the safety of this strain in cats after intramuscular administration. MATERIALS AND METHODS: The ERAG3G strain was rescued in BHK/T7-9 cells using the full-length genome mutated at the amino acid position 333 of the glycoprotein gene of RABV and helper plasmids. Four-week-old mice underwent one or two oral administrations of the ERAG3G strain and were challenged with the highly virulent RABV strain CVSN2c 14 days after the second administration. Clinical symptoms were observed and body weights were measured every day after the challenge. RESULTS: All mice showed complete protection against virulent RABV. In addition, cats intramuscularly inoculated with the ERAG3G strain showed high antibody titers ranging from 2.62 to 23.9 IU/mL at 28-day postinoculation. CONCLUSION: The oral immunization of the ERAG3G strain plays an important role in conferring complete protection in mice, and intramuscular inoculation of the ERAG3G strain induces the formation of anti-rabies neutralizing antibody in cats.

Application of recombinant adenoviruses expressing glycoprotein or nucleoprotein of rabies virus to Korean raccoon dogs

PURPOSE: A new rabies vaccine for animals, including raccoon dogs, in Korea is needed to eradicate rabies infection. In this study, we constructed two recombinant adenoviruses expressing the glycoprotein or nucleoprotein of the rabies virus (RABV). We then investigated the safety and immunogenicity of these strains in raccoon dogs, depending on inoculation route. MATERIALS AND METHODS: Recombinant adenoviruses expressing the glycoprotein (Ad-0910G) or nucleoprotein (Ad-0910N) of rabies were constructed in 293A cells using an adenoviral system. One-year-old raccoon dogs underwent intramuscular (IM) inoculation or oral administration of the recombinant Ad-0910G and Ad-0910N. Clinical symptoms were observed and virus-neutralizing antibodies (VNA) against RABV were measured at 0, 2, 4, and 6 weeks after the immunization. Raccoons were considered positive if VNA titers were > or = 0.1 IU/mL. RESULTS: Raccoon dogs inoculated with the combined Ad-0910G and Ad-0910N virus via the IM route did not exhibit any clinical sign of rabies during the observation period. All raccoon dogs (n = 7) immunized IM had high VNA titers, ranging from 0.17 to 41.6 IU/mL at 2 weeks after inoculation, but 70% (7/10) of raccoon dogs administered viruses via the oral route responded by 6 weeks after administration against RABV. CONCLUSION: Raccoon dogs inoculated with Ad-0910G and Ad-0910N viruses showed no adverse effects. Immunization with the combined Ad-0910G and Ad-0910N strains may play an important role in inducing VNA against RABV in raccoon dogs.

Safety and Immunogenicity of a Recombinant Rabies Virus Strain (ERAG3G) in Korean Raccoon Dogs

A new alternative rabies bait vaccine strain named ERAG3G, which is applicable to wild animals, was developed to eliminate rabies in South Korea. In this study, the safety and immunogenicity of the strain was evaluated in Korean raccoon dogs. The ERAG3G was propagated in BHK/T7-9 cells. Korean raccoon dogs were administered ERAG3G (1 ml, 10(8.0) FAID50/ml) orally or intramuscularly to evaluate its safety and immunogenicity. The raccoon dogs were observed for 70 days after administration, and immunogenicity was measured using a fluorescent antibody virus neutralization test. The ERAG3G strain was not pathogenic to Korean raccoon dogs immunized via the intramuscular or oral route. Raccoon dogs administered the candidate vaccine via the oral route developed high virus neutralizing antibody (VNA) titers ranging from 13.7 to 41.6 IU/ml 70 days post administration. Raccoon dogs inoculated intramuscularly with the ERAG3G strain developed moderate VNA titers ranging from 0.5 to 13.7 IU/ml. These findings suggest that the ERAG3G strain is safe and induces a protective immune response in raccoon dogs.

Seroepidemiological Survey of Aujeszky's Disease Virus in Wild Boar (Sus scrofa) and Raccoon Dogs (Nyctereutes procyonoides koreensis) in Korea

Aujeszky's disease caused by Aujeszky's disease virus (ADV) is one of the most important diseases in the pig industry. In this study, we conducted a seroepidemiological survey of ADV in wild boars and raccoon dogs in South Korea. In total, 217 wild boar sera collected between March and August 2013, and 96 raccoon dogs between 2011 and 2012 were screened for the presence of antibodies against ADV. The sero-positive rates in wild boars and raccoon dogs tested for ADV were found to be 3.55% (8/225) and 0% (0/96), respectively. The presence of virus neutralization antibody titer against ADV means that small number of wild boars was infected with ADV and AD may be circulated continuously in Korean wild boar populations, and that wild boars may act as a potential reservoir of ADV. Therefore, to achieve the declaration of AD free, effective preventive measures to block transmission of AD should be taken to the wild boars.

Strategic model of national rabies control in Korea

Rabies is an important zoonosis in the public and veterinary healthy arenas. This article provides information on the situation of current rabies outbreak, analyzes the current national rabies control system, reviews the weaknesses of the national rabies control strategy, and identifies an appropriate solution to manage the current situation. Current rabies outbreak was shown to be present from rural areas to urban regions. Moreover, the situation worldwide demonstrates that each nation struggles to prevent or control rabies. Proper application and execution of the rabies control program require the overcoming of existing weaknesses. Bait vaccines and other complex programs are suggested to prevent rabies transmission or infection. Acceleration of the rabies control strategy also requires supplementation of current policy and of public information. In addition, these prevention strategies should be executed over a mid- to long-term period to control rabies.

Antibody Response in Cattle and Guinea Pigs Inoculated with Rabies Vaccines

One hundred ninety-five rabies cases in cattle were identified in South Korea since 1993. As most of rabies cases have a relation to rabid Korean raccoon dogs (Nyctereutes procyonoides koreensis), vaccination to animals including cattle is mandatory in rabies risk region. In order to minimize fatal rabies in animals, eradication policy of the disease has been achieved by controlling reservoirs and by mass vaccination. In this study, we compared the antibody response in cattle and guinea pigs inoculated with rabies vaccines commercially available in Korea. Each group of cattle in Gangwon-do was vaccinated intramuscularly with either one of five commercial inactivated vaccines or a live attenuated rabies vaccine (designated as A to F). Serum samples at the time of vaccination and four weeks post vaccination were obtained from the cattle and guinea pigs and were analyzed with virus neutralizing assay (VNA). Each group of cattle inoculating rabies vaccines showed significant virus neutralizing antibody titers (p < 0.05) ranging from 1.55 to 17.8 mean IU/ml compared with the non-vaccinated cattle and guinea pigs inoculated with 1/20 dose of vaccine showed relatively low VN antibody titers ranging from 0.23 to 6.1 mean IU/ml. All cattle immunized with A, C and F showed high VN antibody titers over 0.5 IU/ml and 62.5% and 37.5% of cattle inoculated with D and E showed protective antibody titer, respectively. This finding suggests that the inactivated or live attenuated rabies vaccination commercially available in Korea could induce protective antibody response in Korean cattle, but sero-conversion rate and sero-positive rate showing VN antibody titer over 0.5 IU/ml depend on vaccines.

Serologic Survey of Rabies Virus, Canine Distemper Virus and Parvovirus in Wild Raccoon Dogs (Nyctereutes procyonoides koreensis) in Korea

Oral rabies vaccination (ORV) program for the wild animals in rabies risk regions of Korea has been conducted since 2000. Evaluation of ORV program under field condition and information concerning the incidence of exposure to canine distemper and canine parvovirus (CPV) are needed in wild raccoon dogs (Nyctereutes procyonoides koreensis). Ninety four sera of wild raccoon dogs were screened for antibodies against rabies, canine distemper virus (CDV) and CPV in Korea. The overall prevalence of antibodies against rabies virus (RABV), CDV and CPV in wild raccoon dogs was 35.1%, 89.4% and 24.5%, respectively. Comparisons of sero-prevalences of RABV, CDV and CPV were assayed in two regions (Gyeonggi-do and Gangwon-do). The Gyeonggi-do (36.4%) showed higher sero-positive rate against CPV than Gangwon-do (20.8%). In contrast, Gangwon-do (41.7% and 97.2%) showed higher sero-positive rates against RABV and CDV than Gyeonggi-do (13.6% and 63.6%). These results indicate that there was severe circulation of CDV and CPV among wild raccoon dogs in the two regions of Korea. Furthermore, raccoon dogs showing a protective antibody titer (0.5 IU/ml) were 15.9%, suggesting that new rabies control program such as trap-vaccination-release (TVR) should be launched urgently in rabies risk regions.

Antibody Response in Korean Raccoon Dogs Inoculated with Inactivated Rabies Vaccines

Since sylvatic rabies was first identified in South Korea in 1993, over three million bait vaccine doses have been distributed to rabies risk regions in order to block transmission of rabies among wild animals. New progressive strategy is needed to eliminate sylvatic rabies completely in rabies risk regions. Before applying the preventive program related to eradication, immunogenicity of inactivated rabies vaccines available in Korea has to be evaluated in Korean raccoon dogs (Nyctereutes procyonoides koreensis). Six groups of raccoon dogs in wild rescue center of Gyeonggi-do were vaccinated intramuscularly with single dose of six inactivated commercial rabies vaccines (designated A to F). Serum samples at the time of vaccination, and two and four weeks post vaccination were obtained and analyzed by virus neutralizing assay (VNA). All raccoon dogs inoculated with vaccines C, D, E or F, showed VN antibody titers ranging from 0.5 to 13.77 IU/ml. Half of four raccoon dogs immunized with vaccine B revealed VN titer over 0.5 IU/ml, and one of four raccoon dogs inoculated with vaccine A showed protective antibody titer. This finding suggests that most of the commercially available inactivated rabies vaccines could induce protective immunity in Korean raccoon dogs and be applicable to new rabies control program.

Molecular characterization of Korean rabies virus isolates

The nucleoprotein (N) and glycoprotein (G) of 11 Korean rabies virus (RABV) isolates collected from animals diagnosed with rabies between 2008 and 2009 were subjected to molecular and phylogenetic analyses. Six isolates originated from domestic animals (cattle and dogs) and five were obtained from wild free-ranging raccoon dogs. The similarities in the nucleotide sequences of the N gene among all Korean isolates ranged from 98.1 to 99.8%, while those of the G gene ranged from 97.9 to 99.3%. Based on the nucleotide analysis of the N and G genes, the Korean RABV isolates were confirmed as genotype I of Lyssavirus and classified into four distinct subgroups with high similarity. Phylogenetic analysis showed that the Korean isolates were most closely related to the non-Korean NeiMeng1025B and 857r strains, which were isolated from rabid raccoon dogs in Eastern China and Russia, respectively. These findings suggest that the Korean RABV isolates originated from a rabid raccoon dog in Northeastern Asia. Genetic analysis of the Korean RABV isolates revealed no substitutions at several antigenic sites, indicating that the isolates circulating in Korea may be pathogenic in several hosts.

Effects of alprazolam on capture stress-related serum cortisol responses in Korean raccoon dogs (Nyctereutes procyonoides koreensis)

The purpose of this study was to evaluate the effect of alprazolam on the stress that Korean raccoon dogs (Nyctereutes procyonoides koreensis) may experience while caught in a live trap by measuring their serum cortisol response. The animals were placed in a live trap with or without being pretreated with oral doses of alprazolam. In both groups, pre-trap blood samples were initially collected without anesthesia before the animals were positioned in the live trap; then post-trap blood samples were collected after the animals had remained in the live trap for 2 h. Changes in cortisol levels were observed using a chemiluminescent immunoassay. The level of cortisol increased in the control group and decreased in the alprazolam-pretreatment group (p < 0.05). In this study, we demonstrated that alprazolam pretreatment reduced stress during live trap capture.

Epidemiological Characteristics of Rabies in South Korea from January 2004 to March 2011

Over the seven years from January 2004 to March 2011, 105 rabies cases in three different animal species and one case in human were recorded in South Korea. Forty three (40.6%) cattle and 33 (31.1%) dogs, 29 (27.4%) raccoon dogs were affected and one (0.9%) death in human was reported. The highest annual incidence of rabies was recorded with 27 cases in 2004, and then decreased to a median of 14 cases per year. Eighty cases (76.2%) occurred in Gangwon and 24 cases (22.9%) in Gyeonggi and one case in Seoul Province. All rabies cases occurred in the northern part of the country, namely, Seoul, Gyeonggi and Gangwon Provinces. Since 2007, rabies cases were not reported in Gyeonggi Province, but continued to occur and move eastward in Gangwon Province. The monthly distribution of animal rabies during the seven year period peaked in January, and the incidence was the highest during winter, from December to February, and the least common in summer, from June to September. The epidemiological study indicated that preventive measures including distribution of bait vaccine for the control of rabies in wild animals was helpful to a substantial decrease in number of rabies cases in South Korea.