• Clinical and Experimental Vaccine Research
• /
• 제12권2호
• /
• pp.97-106
• /
• 2023

Purpose: Rabies is a fatal but preventable disease with proper pre-exposure anti-rabies vaccination (ARV). Dogs, as household pets and strays, are the reservoir and vector of the disease, and dog bites have been associated with human rabies cases in Sri Lanka over the past few years. However, other susceptible species having frequent contact with humans may be a source of infection. One such species is sheep and immunity following ARV has never been tested in sheep reared in Sri Lanka. Materials and Methods: We have tested serum samples from sheep reared in the Animal Centre, Medical Research Institute of Sri Lanka for the presence of anti-rabies antibodies following ARV. Sheep serum samples were tested with Bio-Pro Rabies enzyme-linked immunosorbent assay (ELISA) antibody kits used for the first time in Sri Lanka and our results were verified by a seroneutralization method on cells (fluorescent antibody virus neutralization, FAVN test) currently recommended by World Organization for Animal Health and World Health Organization. Results: Sheep received annual ARV and maintained high neutralizing antibody titers in their serum. No maternal antibodies were detected in lamb around 6 months of age. Agreement between the ELISA and FAVN test, i.e., coefficient concordance was 83.87%. Conclusion: Annual vaccination in sheep has an effect on maintaining adequate protection against rabies by measurements of anti-rabies antibody response. Lambs need to be vaccinated earlier than 6 months of age to achieve protective levels of neutralizing antibodies in their serum. Introducing this ELISA in Sri Lanka will be a good opportunity to determine the level of anti-rabies antibodies in animal serum samples.

• 한국동물위생학회지
• /
• 제34권4호
• /
• pp.297-301
• /
• 2011

Rabies virus is transmitted most commonly through a bite from an infected dog. Especially, stray dogs that are not excluded from contact with rabid wild animals can become rabies vectors. Therefore, serological survey of rabies virus from stray dogs in Seoul Metropolitan City was carried out in this study. To investigate prevalence of rabies antibodies in the stray dogs, serum samples were taken from 500 stray dogs between April and December 2010. Antibodies to rabies virus were detected by indirect ELISA. Of 500 tested sera, 147 (29.4%) were positive to rabies virus. Prevalence rates of rabies antibodies(PRRA) in northern and southern Han river region of Seoul were 26.4% and 33.2% respectively. PRRA in male and female dogs were 33.6% and 26.1% respectively. PRRA in less than 1 year, 1~2< years, 2~3< years, 3~5< years, 5~10< years and over 10 years old dogs were 12.7%, 21.6%, 26.4%, 36.4%, 32.5% and 46.4%, respectively. These results indicates that antibody seroprevalence to rabies is still not enough to prevent rabies and rabies vaccination is required to enhance the seroprevalence in the dogs. To improve the situation, measures to raise public awareness of rabies and its prevention is needed. Also, reducing stray dogs and keeping companion animals from contact with wild animals are indispensable for the prevention of rabies.

• 한국동물위생학회지
• /
• 제25권1호
• /
• pp.1-7
• /
• 2002

Recently, the rabies cases have been reported in Paju- and Yangju-gun, Gyeonggj province near Seoul metropolitan area. The Pukhansan national park, nearly located from the cities, is suspected to be a high risk area for incidence and spread of the rabies to metropolitan area. This study was conducted to investigate the prevalence rate for rabies antibody of dogs near the Pukhansan national park and in some other districts in Seoul metropolitan city. From march to october 2001, a total of 306 serum samples were taken from dogs for breeding(189) md pet dogs(117) in 4 districts near the Pukhansan national park and other districts of Seoul. Rabies virus antibodies in sera were detected by neutralizing peroxidase - linked as say (NPLA). Of the 306 sera of dogs tested, 74 (24.2%) were positive to rabies virus antibody. The prevalence rates of rabies antibody in Pukhansan national park area and in the other districts of Seoul city were 23.7% and 25.3%, respectively There was no significant difference in the prevalence rate between these two districts. The prevalence rates of rabies antibody in pet dogs and dogs for breeding were 40.2% and 14.3% respectively. The prevalence rates of rabies antibodies in less than 1 year, 1∼<2 years, 2∼<3 years, and over 3 years old dogs were 14.5%, 22.4%, 32.6%, and 27.1%, respectively, and overall 24.2% in the dog population. In addition, we found that dogs less than 1 year old had lower antibody prevalence than those over 1 year old. It was concluded that enhancement of vaccination is important in the prevention of the rabies, and that rabies vaccines should not be less supplied than the population of the dog.

• 한국동물위생학회지
• /
• 제33권2호
• /
• pp.105-111
• /
• 2010

Rabies virus which belongs to the genus Lyssavirus of the family Rhabdoviridae is known as a highly neurotropic virus and causes fatal encephalitis accompanied by severe neurological symptoms in almost all mammals, including humans. In this study, monoclonal antibodies (MAbs) against rabies virus were produced, characterized and applications of MAbs as diagnostic reagents were assessed Spleen and inguinal lymph node cells from Balb/c mouse immunized with purified rabies virus were fused with SP2/O myeloma cells using polyethylene glycol (PEG) and hybridoma cells producing rabies virus-specific MAbs were screened by an indirect fluorescent antibody test. A total of ten MAbs were produced against rabies virus. The protein specificity and neutralizing activity of MAbs were determined by Western blot analysis and fluorescent antibody virus neutralization test, respectively. As a result, two MAbs, 5G3 and 6H4 had specificity for nucleoprotein (N protein) and two other MAbs, 5B1 and 5C1 had neutralizing activity for rabies virus. Some MAbs recognized the rabies virus-infected bovine brain stem cells by immunohistochemistry (IHC) assay. In conclusion, it was confirmed that MAbs produced in this study were rabies virusspecific and could be used as reliable diagnostic reagents for the detection of rabies virus.

• 한국미생물·생명공학회지
• /
• 제46권3호
• /
• pp.269-276
• /
• 2018

Rabies virus (RABV)-specific antibodies in animals and humans are measured using standard methods such as fluorescent antibody virus neutralization (FAVN) tests and rapid fluorescent focus inhibition tests, which are based on cell culture systems. An alternative assay that is safe and easy to perform is required for rapid sero-surveillance following mass vaccination of animals. Two purified monoclonal antibodies (4G36 and B2H17) against RABV were selected as capture and detection antibodies, respectively. A genetically modified RABV, the ERAGS strain, was propagated and concentrated by polyethylene glycol precipitation. Optimal conditions for the RABV antigen, antibodies, and serum dilution for a blocking enzymelinked immune sorbent assay (B-ELISA) were established. We evaluated the sensitivity, specificity, and accuracy of the B-ELISA using serum samples from 138 dogs, 71 raccoon dogs, and 25 cats. The B-ELISA showed a diagnostic sensitivity of 95.8-96.3%, specificity of 91.3-100%, and accuracy of 96.0-97.2% compared to the FAVN test. These results suggest that the B-ELISA is useful for sero-surveillance of RABV in dogs, raccoon dogs, and cats.

• 대한수의학회지
• /
• 제63권2호
• /
• pp.13.1-13.6
• /
• 2023

Animals imported from abroad are a cause of rabies outbreaks in many countries. Therefore, rabies serology testing for dogs and cats traveling abroad is an important measure to reduce the incidence of rabies. Rabies virus antibodies were measured in sera collected from 2,367 dogs and 894 cats between 2017 and 2021. A serum sample with a value of 0.5 IU/mL or higher was considered a pass. The overall pass rates for rabies virus were 96.4% in dogs and 98.4% in cats. The mean rabies virus neutralization assay titers were higher in cats than in dogs and in female than in male animals. According to age, 6-year-old dogs and 9-year-old cats had the highest virus neutralization assay titers. Of the failure cases, 53.0% (53/100) were dogs or cats less than 1 year old. Although the average failure rates in dogs and cats were low at 3.5% and 1.6%, respectively, the factors influencing failure were age and vaccine manufacturer. Therefore, it is necessary to observe the vaccination interval and timing of blood collection after boosting.

• 한국동물위생학회지
• /
• 제24권2호
• /
• pp.109-116
• /
• 2001

This study was conducted to investigate the prevalence rate for rabies antibody (PRRA) of dogs near the Pukhansan National Park and in some other districts in Seoul Metropolitan city. From April to July 2000, a total of 414 serum samples were taken from dogs for breeding (92), pet dogs (162), and unclaimed/stray dogs (162). Rabies virus antibodies were detected by neutralizing peroxidase-linked assay (NPLA). Of 414 sera tested, 145 (35%) were positive to rabies virus antibody. PRRA in Pukhansan National Park area and in the other districts of Seoul city were 34.8% and 35.4%, respectively. There was no significant difference in the prevalence rate between these two districts. PRRA in pet dogs, unclaimed/stray dogs, and dogs for breeding were 39.5%, 35%, and 27.2% respectively. PRRA in dogs from residential areas, apartments, animal hospitals, and farms were 32.5%, 60%, 35.3%, and 26.7% respectively. Especially, the dogs reared in apartments had a significantly higher seroprevalence (60%) than those in residential or farm areas. PRRA in less than 1 year, 1~<2 years, 2~<3 years, and over 3 years old dogs were 14.7%, 40.4%, 38.4%, and 53% respectively, and overall 35% in the dog population. In addition, we found that dogs less than 1 year of age had lower antibody prevalence than those over 1 year of age. It was concluded that enhancement of vaccination is important in the prevention of the rabies, and that rabies vaccines should not be less supplied than the population of the dog.

• Journal of Microbiology and Biotechnology
• /
• 제27권4호
• /
• pp.718-724
• /
• 2017

The combination of rabies immunoglobulin (RIG) with a vaccine is currently effective against rabies infections, but improvements are needed. Genetic engineering antibody technology is an attractive approach for developing novel antibodies to replace RIG. In our previous study, a single-chain variable fragment, scFv57R, against rabies virus glycoprotein was constructed. However, its inherent weak stability and short half-life compared with the parent RIG may limit its diagnostic and therapeutic application. Therefore, an acidic tail of synuclein (ATS) derived from the C-terminal acidic tail of human alpha-synuclein protein was fused to the C-terminus of scFv57R in order to help it resist adverse stress and improve the stability and half-life. The tail showed no apparent effect on the preparation procedure and affinity of the protein, nor did it change the neutralizing potency in vitro. In the ELISA test of molecular stability, the ATS fusion form of the protein, scFv57R-ATS, showed an increase in thermal stability and longer half-life in serum than scFv57R. The protection against fatal rabies virus challenge improved after fusing the tail to the scFv, which may be attributed to the improved stability. Thus, the ATS fusion approach presented here is easily implemented and can be used as a new strategy to improve the stability and half-life of engineered antibody proteins for practical applications.

• 대한수의학회지
• /
• 제57권4호
• /
• pp.209-214
• /
• 2017

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 sero-surveillance 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.

• Journal of Plant Biotechnology
• /
• 제34권4호
• /
• pp.339-346
• /
• 2007

식물화분발아는 수분수정을 통한 식물의 종자형성과 식물개체의 지속적 세대유지를 위한 중요한 요소이다. 따라서, 본 연구에서는 boric acid를 이용하여 형질전환 하지 않은 (NT) 식물과 항암활성 (anti-colorectal cancer mAb CO17-1A, anti-breast cancer mAb BR55) 혹은 항바이러스 활성을 갖는 항체 (anti-rabies virus mAb57)의 유전인자를 발현하는 형질전환식물 간의 in vitro pollen germination과 Pollen tube growth의 차이를 알아보았다. Boric acid의 화분발아에 미치는 영향을 보기 위하여 0, 5, 10, 15, 20, $40{\mu}g/mL$의 농도를 갖는 발아용액 (germination buffer)을 담배식물 화분에 처리하여 pollen germination rate을 본 결과 $20{\mu}g/mL$에서 가장 높게 발아율 (49.5%)을 보였다. Boric acid ($20{\mu}g/mL$)에 대해 형질전환식물과 NT 식물에서 화분발아율을 관찰한 결과, boric acid를 처리하지 않은 화분에 비해 형질전환여부에 관계없이 발아율이 3-10배 증가하였다. 또한, boric acid $20{\mu}g/mL$농도에서 형질전환식물과 NT 식물의 pollen tube length의 성장률이 가장 높게 나타났다. 시간 별로 형질전환 식물들의 Pollen tube length를 확인 한 결과, germination buffer ($20{\mu}g/mL$)를 처리하고 난 후 3시간까지 가장 빠른 성장률을 보였으며, 3시간 후에는 그 성장률이 둔화됨을 확인하였다. Western blot를 이용하여 식물의 화분과 잎에서 황체유전자 단백질 발현을 확인하였다. 화분과 잎에서의 발현을 비교해 본 결과, 잎에서 항체의 안정한 발현을 확인하였다. 본 연구결과를 통해 $20{\mu}g/mL$의 boric acid가 항체발현 형질전환담배식물의 in vitro pollen germination을 보기 위해 가장 적절한 농도라는 것을 제시하였다. 궁극적으로 의료용 항체유전자를 발현하는 형질전환식물의 in vitro pollen germination viability를 확인함으로써 형질전환식물의 화분의 안정적인 수분수정이 이루어질 수 있음을 간접적으로 제시하였다.