• Title, Summary, Keyword: rabbit hemorrhagic disease

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Rabbit Hemorrhagic Disease Virus Variant Recombinant VP60 Protein Induces Protective Immunogenicity

  • Yang, Dong-Kun;Kim, Ha-Hyun;Nah, Jin-Ju;Song, Jae-Young
    • Journal of Microbiology and Biotechnology
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    • v.25 no.11
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    • pp.1960-1965
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    • 2015
  • Rabbit hemorrhagic disease virus (RHDV) is highly contagious and often causes fatal disease that affects both wild and domestic rabbits of the species Oryctolagus cuniculus. A highly pathogenic RHDV variant (RHDVa) has been circulation in the Korean rabbit population since 2007 and has a devastating effect on the rabbit industry in Korea. A highly pathogenic RHDVa was isolated from naturally infected rabbits, and the gene encoding the VP60 protein was cloned into a baculovirus transfer vector and expressed in insect cells. The hemagglutination titer of the Sf-9 cell lysate infected with recombinant VP60 baculovirus was 131,072 units/50 μl and of the supernatant 4,096 units/50 μl. Guinea pigs immunized twice intramuscularly with a trial inactivated RHDVa vaccine containing recombinant VP60 contained 2,152 hemagglutination inhibition (HI) geometric mean titers. The 8-week-old white rabbits inoculated with one vaccine dose were challenged with a lethal RHDVa 21 days later and showed 100% survival rates. The recombinant VP60 protein expressed in a baculovirus system induced high HI titers in guinea pigs and rendered complete protection, which led to the development of a novel inactivated RHDVa vaccine.

Pathological observations of rabbit hemorrhagic disease (토끼 출혈병의 병리학적 관찰)

  • 문운경;하준일;인민권;김순복;곽수동
    • Korean Journal of Veterinary Service
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    • v.23 no.4
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    • pp.309-312
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    • 2000
  • Pathological findings of natural cases of rabbit hemorrhagic disease was investigated. Clinically inappentence, increase in body temperature, depression, bloody foam from nostrils, and sudden death were recognized. Characteristic anatomical findings were hemorrhages in the lungs, liver, kidneys, and heart. Intestinal catarrh and retention of turbid urine in urinary bladder were also often observed. Severe .necrosis of hepatic cells, massive hemorrhages in many organs and membranous glomerulonephritis with hyaline droplet formation were characteristic changes under the microscopy. Perivascular cuffing of brain and catarrhal enteritis were also seen in many cases.

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Sequential pathologic changes and viral distribution in rabbits experimentally infected with new Korean strain of rabbit hemorrhagic disease virus (RHDVa) (새로운 국내 분리 토끼출혈병바이러스(RHDVa)를 감염시킨 토끼에서의 경시적인 병리학적 변화와 조직 내 바이러스 항원 분포)

  • Park, Jung-Won;Chun, Ji-Eun;Yang, Dong-Kun;Bak, Eun-Jung;Kim, Han;Lee, Myeong-Heon;Hwang, Eui-Kyung;Lee, Chung-Bok;Woo, Gye-Hyeong
    • Korean Journal of Veterinary Research
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    • v.52 no.2
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    • pp.125-131
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    • 2012
  • Rabbit hemorrhagic disease is a highly acute and fatal viral disease caused by rabbit hemorrhagic disease virus (RHDV). Since first outbreak in Korea 1987, RHDV has been continually affected in the country, but the pattern of outbreak seem to be changed. In this study, to understand the pathogenesis of the new RHDVa serotype, we therefore carried out to inoculate RHDVa to rabbits, and to examine the sequential histopathologic changes and viral distribution. Macroscopically, various sized dark red or white spots or appearance were observed in the liver, lung, kidney uterus and ureter. In euhanized rabbits, significant pathologic findings such as infiltration of heterophils and mononuclear cells were observed at 24 hours after inoculation (HAI), and these were sequentially extended periportal to centrilobular area. However, in dead rabbits, severe hepatic degeneration and/or necrosis with relatively weak inflammatory responses were observed. RHDV antigens began to detect in liver, spleen, and lung from 12 HAI by PCR. Immunohistochemically, RHDV positive cells were seen in only liver from 24 HAI, and the degree of immunogen reactivity was stronger in dead rabbits than in euthanized ones. In conclusion, RHDVa caused the subacute or chronic infection accompanying low mortality and moderate to severe inflammatory reaction in rabbits, suggesting the possibility that RHD could become endemic.

Studies on the rabbit viral hepatitis I. Electron microscopic observation of the acute hepatic lesions in experimentally infected rabbit (토끼의 바이러스성(性) 간염(肝炎)에 관(關)한 연구(硏究) I. 실험적(實驗的) 감염토(感染兎)의 급성간염조직(急性肝炎組織)의 전자현미경적(電子顯微鏡的) 관찰(觀察))

  • Lee, Cha-soo;Park, Cheong-kyu
    • Korean Journal of Veterinary Research
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    • v.29 no.4
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    • pp.531-540
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    • 1989
  • A new sudden death in rabbits appeared in China and Korea in 1984 and 1985, respectively, and was recognized to be an acute infectious disease caused by a virus. The disease was reported as a "new viral disease," and thereafter, a tentative name of "viral hemorrhagic disease", "hemorrhagic pneumonia" or "viral hemorrhagic pneumonia" has been described in the case reports. But authors had called the viral disease "rabbit viral hepatitis" due to picornavirus infection, because the principal lesion of the disease was an acute hepatitis. The purpose of this report is to describe the electron microscopic findings on the livers in experimentally infected rabbits. All the livers of the affected rabbits were shown to have degenerative changes of a type that is characteristic of acute hepatitis. In the liver cells, there were dilation of rER and mitochondria, vacuole formation of various sizes, and appearances of many virus-like particles in the vicinity of rER, granular bodies and crystalline arrays of viral particles in the cytoplasm with necrotic changes of the nucleus. Clusters of virus-like particles and viral crystals appeared in the cytoplasm of sinusoid endothelial cells and Kupffer's cells with morphological changes of organelles. Also viral crystals were demonstrated in the cytoplasm of macrophages among the liver cells. On the whole, the liver cells had many virus-like particles and a few crystalline arrays of viral particles. Therefore, this implies that the liver cells are the main site of the viral replication in inducing the viremia. It was concluded that the liver was the primary target organ of this viral disease, and the pathological and the ultrastructural evidence suggest that the virus may be belong to genus enterovirus.

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Hematological and blood chemical changes of rabbits infected with viral hemorrhagic disease virus (토끼 출혈성 바이러스에 감염된 토끼의 혈액상과 혈액화학치의 변화)

  • Yoon, In-joong;Jeon, Yun-seong
    • Korean Journal of Veterinary Research
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    • v.30 no.1
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    • pp.73-78
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    • 1990
  • Hematological and blood chemical changes of rabbits infected with rabbit hemorrhagic disease virus were studied and the results were summarized as follows: 1. Total leukocyte count ($2,410{\pm}1,076/{\mu}l$), lymphocyte count ($1,582{\pm}632.5{\mu}l$) and heterophil count ($705{\pm}411.1{\mu}l$) were significantly decreased after 24 hours of the infection (p<0.01). However, no significant changes were observed in monocyte, eosinophil and basophil numbers. 2. A significant increase of aspartate aminotransferase (96IU/L), alanine aminotransferase (96IU/L) and alkaline phosphatase ($401.1{\pm}131.8IU/L$) was observed (p<0.01). 3. A moderate increase of BUN ($26.9{\pm}3.6mg/100ml$) and creatinine ($3.2{\pm}1.9mg/100ml$) was observed (p<0.05). 4. No significant changes of r-GTP, thymol turbidity, glucose, cholesterol, albumin, total serum protein, fibrinogen were observed.

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Identification and molecular characterization of a rabbit hemorrhagic disease virus variant (KV0801) isolated in Korea

  • Yang, Dong-Kun;Kim, Byoung-Han;Lee, Kyung-Woo;Kim, Ji-Yeon;Kim, Hee-Jin;Choi, Sung-Suk;Chun, Ji-Eun;Son, Seong-Wan
    • Korean Journal of Veterinary Research
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    • v.49 no.3
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    • pp.207-213
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    • 2009
  • Rabbit hemorrhagic disease (RHD) is caused by RHD virus (RHDV) and is one of the most fatal diseases of rabbits. Acute death of rabbits occurred in a farm located in the Gyeonggi province of South Korea. The virus was isolated and confirmed as RHDV based on reverse transcription polymerase chain reaction and hemagglutination assay (HA), and the isolate was designated as KV0801. The nucleotide sequence of the complete VP60 gene of KV0801 was determined and the corresponding amino acid sequence was deduced. Molecular analysis showed that the KV0801 isolate can be classified as a pandemic antigenic variant strain, RHDVa. The VP60 nucleotide sequence and deduced amino acid homology between KV0801 and other Korean isolate, RHF89, which was isolated in 1988, were 92.1 and 94.3%, respectively. The pathogenicity of the KV0801 isolate at an HA titer ranging from 16,384 to 0.16 HA units was evaluated in five-month-old SFP rabbits. The rabbits inoculated with KV0801 isolate containing more than 1.63 HA units died within six days of inoculation. These results suggest that a highly pathogenic RHDVa is circulating in the rabbit populations of Korea.

Pathogenicity, hemagglutinability, and the effect of physicochemical agents on virus of rabbit hemorrhagic disease (토끼 출혈성 바이러스의 병원성, 적혈구응집성 및 물리화학적 요인에 대한 영향)

  • Yoon, In-joong;Jeon, Yun-seong
    • Korean Journal of Veterinary Research
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    • v.30 no.1
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    • pp.65-71
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    • 1990
  • Rabbits were experimentally infected with rabbit hemorrhagic disease virs and the viral pathogenicity, hemagglutinability, and the effect of physicochemical factors were studied. The experimental results were summariaed as follows: 1. Mean rectal temperature of 11 infected rabbits was $40.0{\pm}0.47^{\circ}C$ prior to the virus inoculation, and $39.9{\pm}0.75^{\circ}C$ after 12hrs., $40.2{\pm}0.65^{\circ}C$ after 24hrs., $40.1{\pm}0.77^{\circ}C$ after 36hurs, and $40.6{\pm}0.56^{\circ}C$ just before the death. 2. Mean death time of infected rabbits was $40.3{\pm}22.0$ honrs and its range was 24 to 93 hours. 3. O, B, AB and A type of human erythrocytes were shown their HA in the order, but rabbit and chicken erythrocytes were not hemagglutinated by the virus. 4. In the hemagglutination, less than 0.25 per cent of a final concentration of erythrocytes and 0.2 per cent of BSA in PBS resulted in the best hemagglutination. Phosphate concentration in a range of 0.01M to 0.10M in PBS was not influenced on the hemagglutination reaction, and its pH 7.0 resulted in a better HA. 5. The hemagglutinating titers, in log 2 scale, of organs and tissues of the virus infected rabbits were $9.3{\pm}3.8$ (liver), $9.1{\pm}3.9$ (blood), $6.2{\pm}2.6$ (spleen) and $5.0{\pm}2.5$ (kidney). 6. The physicochemical factors such as heating ($50^{\circ}C$, 10 min.), trypsin treatment (0.05 pre cent, 5 min.), acid treatment (pH 3.0, 20 min.) and ether extraction (3 times) were not affective to the stability of virus and viral HA activities.

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Rapid and Easy Diagnosis of Rabbit Haemorrhagic Disease by In Situ Hybridization (In situ Hybridization에 의한 토끼출혈증(rabbit haemorrhagic disease)의 신속.간편한 진단)

  • Park, Nam-Yong;Cho, Ho-Seong;Cho, Kyoung-Oh;Kim, Sang-Jip;Park, Hyung-Seon
    • Korean Journal of Veterinary Pathology
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    • v.5 no.2
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    • pp.57-62
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    • 2001
  • Recently various molecular diagnostic techniques have been used to identify rabbit hemorrhagic disease virus (RHDV), a causative agent responsible for acute hepatitis and disseminated intravascular coagulation in rabbit. But they were hard to perform and time consuming. To detect RHDV in a rapid and easy way, we developed biotinylated oligonucleotide probe within ORF 1 region encoding the polyprotein of RHDV in formalin-fixed and paraffin-embedded tissues from various tissues of 20 rabbits naturally infected with RHDV, Our in situ hybridization (ISH) was quickly carried out within two hours by MicroProbe capillary action system. The ISH produced a positive reaction in liver, kidney and lung. In conclusion, ISH with a biotintlated oligonucleotide probe provided a useful diagnostic method for detecting RHDV.

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Sequential hepatic ultrastructural changes and apoptosis in rabbits experimentally infected with Korean strain of rabbit hemorrhagic disease virus (RHDVa) (국내 분리 토끼출혈병 바이러스(RHDVa)를 감염시킨 토끼 간장에서의 경시적인 초미세구조 변화와 apoptosis)

  • Park, Jung-Won;Chun, Ji-Eun;Bak, Eun-Jung;Kim, Han;Lee, Myeong-Heon;Hwang, Eui-Kyung;Kim, Jae-Hoon;Lee, Chung-Bok;Woo, Gye-Hyeong
    • Korean Journal of Veterinary Research
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    • v.53 no.1
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    • pp.11-17
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    • 2013
  • In this study, to understand the pathogenesis of new rabbit hemorrhagic disease virus (RHDVa) serotype, we carried out to administrate RHDVa to rabbits, and to examine sequential electron microscopic changes and relationship between pathogenesis and apoptosis. TUNEL-positive cells began to be observed from 24 hours after inoculation (HAI) and the number of positive cells was slightly increased with the course of time. Whereas marked increase of positive cells was seen in the liver from the rabbits died acutely. Typical viral particles with cup-like projections and a diameter of 30~40 nm were detected in homogenized liver samples and tissues at 36 and 48, and 48 HAI, respectively. Ultrastructurally, glycogen deposition was observed from the first stage of hepatocellular degeneration by RHDVa infection and then, swelling and disruption of cristae of mitochondria by viral particles, swelling of smooth endoplasmic reticulum, vacuoles and vesicles were detected. Condensation, margination and fragmentation of chromatin were observed in degenerative hepatocytes at 36 and 48 HAI, indicating apoptotic bodies. These data offer that hepatocytic apoptosis by RHDV infection could be closely related with mitochondrial impairment in the hepatocytes.

Cerebral lesions of encephalitozoonosis in the rabbits died of rabbit hemorrhagic disease (토끼 출혈병(出血病)으로 죽은 토끼의 대뇌(大腦)에서 보인 encephalitozoonosis의 병변(病變))

  • Park, Jae-hak;Lee, Yong-soon;Itakura, C.
    • Korean Journal of Veterinary Research
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    • v.32 no.2
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    • pp.227-233
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    • 1992
  • Out of twenty rabbits which died of rabbit hemorrhagic disease spontaneously occurring in Korea, five animals had a concurrent infection with Encephalitozoon cuniculi in the cerebrum. The lesions were composed of granulomas, leptomeningitis and perivascular cuffing with mononuclear cells. The granulomas consisted of a central necrotic focus surrounded by an infiltration with plasma cells, lymphocytes and macrophages. Gliosis was associated with the granulomas. Gram-positive organisms were detected in the cerebrum from two rabbits. They were oval to rod-shaped with blunt round ends. The distribution of the pathogens was investigated by the direct avidin-biotin peroxidase complex method. They were present in pseudocysts and macrophages. Pseudocysts were found in the granulomas as well as the neuropil without cellular reactions. Some organisms were present within reticulo-endothelial cells of blood capillaries and macrophages in the subarachnoid spaces. These organisms had ultrastructural characteristics consistent with Encephalitozoon cuniculi.

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