Hepatic lesions in young rabbits experimentally infected with rabbit haemorrhagic disease virus

Twenty young rabbits (eleven 2-week-old and nine 4-week-old) were experimentally infected with rabbit haemorrhagic disease virus (RHDV) to clarify susceptibility. They were killed chronologically up to 96 hours post-inoculation (PI) and examined for lesions. All inoculated rabbits were clinically normal, but grossly minute white or grey spots were detected throughout the liver. Histologically, the lesions consisted of aggregates of lymphocytes, macrophages and heterophils, with or without acidophilic bodies and necrotic hepatocytes. Immunohistochemically, RHDV antigens were found in the degenerated hepatocytes and in macrophages. The cellular aggregates were considered to be a reaction to necrotic hepatocytes infected with RHDV. It was concluded that some hepatocytes are susceptible to RHDV in young rabbits.     

Typing of rabbit haemorrhagic disease virus from New Zealand wild rabbits

Abstract

AIMS: To examine the gestation lengths and occurrence of daytime foaling of Standardbred mares foaling outdoors at stud farms in Southland, New Zealand (latitude 45?S).

METHODS: Data were collected prospectively at two commercial Standardbred stud farms (Farms A and B), during the 2008/9 and 2009/10 breeding seasons (October to February). For each foaling, the identity of the mare, date and time of foaling, gender of foal, time the mare passed the fetal membranes, time the foal stood, and foaling problems including dystocias, were recorded. The effect of farm, season, gender of foal, month of artificial insemination (AI) or foaling, age and parity of mare on gestation length, percentage of mares foaling during daylight hours, and percentage of foalings recorded as dystocia, were examined.

RESULTS: A total of 614 foaling records were obtained from 507 mares. For 594 foalings with complete records, mean gestation length was 349 (SE 0.5) days. Mean gestation length was shorter for fillies (347.8 (SE 0.6) days) than colts (350.3 (SE 0.6) days) p = 0.021) and decreased with month of AI, from October to February (p = 0.001). The time of foaling was bimodally distributed with the primary peak around 0200 hours and a secondary smaller peak around 1300 hours. The percentage of mares foaling in daylight was lower on Farm A (69/285 (24%)) than Farm B (128/313 (41%)) (p = 0.001). Colt foals were less likely to be born during the day than fillies (OR = 0.63; (95% CI = 0.44–0.88); p = 0.008), but there was no effect of age or parity of mare or month of foaling (p>0.05).

CONCLUSIONS AND CLINICAL RELEVANCE: Mean gestation length of mares in this study was longer than that previously reported from other countries. Longer gestation length decreases the time available for these mares to get back into foal in the same season. It is important that managers take this into account and ensure optimum conditions for conception; breeding at the first postpartum oestrus may be essential. There were more mares foaling in daylight hours on one stud than has previously been reported. Under these conditions mares should be monitored for foaling during daytime as well as at night.     

Viral haemorrhagic disease in rabbits: A review

An acute haemorrhagic disease of rabbits was first reported in a southern province of China in 1984. It subsequently spread rapidly to South China and some parts of North China. The disease is characterized by an acute onset with fever, rapid respiration and sudden death. There is a high morbidity and mortality rate. The pathological changes are consistent with severe generalized circulatory dysfunction (hyperaemia, congestion and haemorrhage), marked degeneration of parenchymatous tissue, pronounced serous-haemorrhagic pneumonia and extensive disruption of reticulo-lymphoid tissue. The disease has been named rabbit viral haemorrhagic disease and it has been suggested that the aetiological agent is a picornavirus. A tissue-derived vaccine has been prepared by homogenizing the liver, lung, spleen and kidney of infected rabbits and inactivating with formaldehyde. This review summarizes the information on the aetiology, epidemiology and clinical and pathological aspects of this new rabbit disease.     

Immunohistological diagnosis of rabbit haemorrhagic disease in experimentally infected rabbits in Spain.

The immunoreactivity of routinely processed liver and lung tissue samples obtained from rabbits inoculated with tissue explants from naturally infected animals when antisera directed against parvovirus from different species (canine, feline and porcine) as well as a RHD virus antiserum were employed has been tested by different immunoperoxidase methods. Cross-reactivity between RHD-virus antigens and parvovirus antigens was present. Best results were obtained with RHD and canine parvovirus antisera with the ABC method. The immunoreactivity in the liver was found in hepatocytes, Kupffer and bile duct cells. In the lung, it was exclusively observed in intravascular macrophages.     

Phagocytosis of neutrophils in rabbits infected with antigenic variants of RHD (rabbit haemorrhagic disease) virus

The present study was aimed at determining changes in chosen elements of phagocytosis in rabbits infected with 3 antigenic variants of RHD - Hartmannsdorf, Pv97 and 9905, which differed in haemagglutination ability. The animals were tested for phagocytosis parameters, and the results revealed that the examined strains showed the differences. These variations regarded mainly Pv97 strain, as the intensity of the changes were 5 times stronger in comparison to strain Hartmannsdorf and 9905. As all of the strains examined are signified as antigenic variants, we have stated that this feature does not determine their immunological picture. The results suggest the existence of immunological dissimilarities among strains of the RHD virus, which was revealed for the first time in antigenic variants.     

Crypt abscesses in the caeca of feral rabbits in a rabbit haemorrhagic disease endemic region of New Zealand

AIM: To describe the pathology of crypt abscesses in the caeca of feral rabbits in the Manawatu region of New Zealand, and to examine the possible relationship between their prevalence and rabbit haemorrhagic disease (RHD) virus (RHDV) infection.

METHODS: During the course of a 3-year study of RHD, 173 wild rabbits (Oryctolagus cuniculus) were shot on three pastoral livestock farms in the Manawatu region of New Zealand. All rabbits were necropsied, and tissue samples of the caeca were examined histologically. The age of each animal was determined, and blood samples collected for the detection of RHDV antibodies. Logistic regression was used to model the odds of rabbits having crypt abscesses.

RESULTS: At necropsy, 63/173 (36.4%) rabbits were found to have small circular black nodules on the mucosal surface of their appendix caecum and/or sacculus rotundus. Microscopically, these were identified as small crypt abscesses composed of dilated sacs at the base of the mucosa that were often lined by a thin layer of attenuated epithelial cells. They usually contained large amounts of concentrically-laminated mucopolysaccharide material that was sometimes pigmented, inflammatory debris, and were often the site of a moderate multifocal appendicitis. Although RHDV was active in the study area, no association was found between RHDV antibodies in serum and the presence of lesions. The lesions were more common in older individuals and those born in summer or autumn.

CONCLUSIONS: This is the first report of crypt abscesses with inflammation and necrotic debris in the caeca of rabbits. No association between the occurrence of crypt abscesses and RHDV infection was identified. Wild rabbits born in a particular season were presumably exposed very early in life to conditions that caused the crypt abscesses to develop. Alternatively, association with season of birth may represent rabbits that were of similar age in a later stage of their lives, when they became exposed to the cause of the lesions, which remains unidentified.     

Crypt abscesses in the caeca of feral rabbits in a rabbit haemorrhagic disease endemic region of New Zealand

AIM: To describe the pathology of crypt abscesses in the caeca of feral rabbits in the Manawatu region of New Zealand, and to examine the possible relationship between their prevalence and rabbit haemorrhagic disease (RHD) virus (RHDV) infection. METHODS: During the course of a 3-year study of RHD, 173 wild rabbits (Oryctolagus cuniculus) were shot on three pastoral livestock farms in the Manawatu region of New Zealand. All rabbits were necropsied, and tissue samples of the caeca were examined histologically. The age of each animal was determined, and blood samples collected for the detection of RHDV antibodies. Logistic regression was used to model the odds of rabbits having crypt abscesses. RESULTS: At necropsy, 63/173 (36.4%) rabbits were found to have small circular black nodules on the mucosal surface of their appendix caecum and/or sacculus rotundus. Microscopically, these were identified as small crypt abscesses composed of dilated sacs at the base of the mucosa that were often lined by a thin layer of attenuated epithelial cells. They usually contained large amounts of concentrically-laminated mucopolysaccharide material that was sometimes pigmented, inflammatory debris, and were often the site of a moderate multifocal appendicitis. Although RHDV was active in the study area, no association was found between RHDV antibodies in serum and the presence of lesions. The lesions were more common in older individuals and those born in summer or autumn. CONCLUSIONS: This is the first report of crypt abscesses with inflammation and necrotic debris in the caeca of rabbits. No association between the occurrence of crypt abscesses and RHDV infection was identified. Wild rabbits born in a particular season were presumably exposed very early in life to conditions that caused the crypt abscesses to develop. Alternatively, association with season of birth may represent rabbits that were of similar age in a later stage of their lives, when they became exposed to the cause of the lesions, which remains unidentified.     

Serology of rabbit haemorrhagic disease virus in wild rabbits before and after release of the virus in New Zealand

Rabbit haemorrhagic disease virus (RHDV) was illegally released in New Zealand in August 1997. The initial release and spread of the virus was conducted by landholders in an effort to reduce costs associated with more conventional control methods (poisoning and shooting). Serum was collected from wild rabbits throughout the Otago region prior to the release and from 13 sites in the months following the first epizootic. Following the occurrence of the first RHDV epizootic on 13 pastoral farming properties a range of survival rates was found. The major factor influencing the survival rate was found to be the method of release, with widespread use of carrot or oat baits containing RHDV resulting in poor kills. Widespread use of baits also resulted in higher levels of antibody in surviving adult rabbits with a higher proportion of adult females surviving the epizootic, compared with properties where the disease was allowed to spread naturally. A correlation was found between survival rate and the percentage of surviving adults with high levels of antibody. These results suggest that poor kill rates are not due to poor spread of the virus, that the large-scale use of baits resulted in protective immunisation and that rabbit control should in the future be achieved through establishing naturally spreading epidemics rather than widespread use of baits.     

Apoptosis of peripheral blood leukocytes from rabbits infected with non-haemagglutinating strains of rabbit haemorrhagic disease virus (RHDV)

The report demonstrates that the induction of apoptosis in peripheral blood granulocytes and lymphocytes of rabbits infected with three non-haemagglutinating RHDV strains (English Rainham, German Frankfurt, and Spanish Asturias) is a crucial determinant of the pathogenesis of rabbit haemorrhagic disease. Apoptosis was measured by flow cytometric detection of caspase activity. These studies demonstrated that the investigated RHDV (rabbit haemorrhagic disease virus) viral strains affected leukocyte apoptosis to varying degrees. Enhanced leukocyte apoptosis was detected between 4 and 36h after infection and was more pronounced in lymphocytes than in granulocytes. The data presented here thus provide a preliminary understanding of the kinetics of apoptosis in leukocytes of rabbits infected with RHDV.     

Immunohistochemical localisation of rabbit haemorrhagic disease virus VP-60 antigen in early infection of young and adult rabbits

This study evaluated the time course distribution of rabbit haemorrhagic disease virus (RHDV) structural protein VP60 in tissues from experimentally infected rabbits from three different age groups. Viral VP60 antigen could not be detected in tissue samples from animals under four weeks, and only a few hepatocytes (0.01 to 0.2 per cent) were stained in the 6-week-old animals. A 6-week-old rabbit euthanised at 72 hpi showed VP60-labelling in hepatocytes and macrophages close to areas of inflammation. Viral VP60 antigen was detected as early as 12 hpi in a few hepatocytes (0.03 per cent) from adult animals. Within this age group, the extent of hepatocyte labelling considerably increased at 18 (3.0 per cent), 24 (25.5 per cent), 36 (50 per cent) and 48 (60 per cent) hpi. Extrahepatic viral VP60 antigen was also detected at 36 and 48 hpi in spleen macrophages and lymphocytes from adult rabbits. These findings support the hypothesis that the hepatocyte is the only cell type in the liver able to support RHDV replication almost immediately after viral infection.     

Etiology of rabbit haemorrhagic disease spontaneously occurring in Korea.

Causative agent of rabbit haemorrhagic disease (RHD) was purified by CsCl density gradient centrifugation from the liver homogenate of rabbits infected with RHD virus which originated from Korea. The viral particles were 35-40 nm in diameter, and had hollow depressions on their surface. Protein A-gold immunoelectron microscopy clearly showed that the convalescent antisera of diseased rabbits reacted specifically with the virus particles. SDS-PAGE and Western blot analyses demonstrated that the structural protein of the virus was composed of a single major polypeptide of 63 kD. These findings indicate that the causative agent of RHD, tentatively named as picornavirus in Korea, belongs to calicivirus.     

Immunohistological diagnosis of rabbit haemorrhagic disease (RHD).

In the present study the diagnostic use of a biotinylated serum from an immune rabbit was investigated by means of an Avidin-Biotin-Complex (ABC)-Peroxidase method on paraffin sections. 15 cases of RHD which had been verified histologically and/or by haemagglutination test (HA), 4 suspected cases and 3 cases without history of RHD were included (cases 1 to 22). From 5 prospective cases a wider tissue range was examined (cases 23 to 25 and 29 to 30). Furthermore lungs, liver and placenta of 3 fetuses from a RHD affected dam were investigated (cases 26 to 28). The 20 typical cases had intense intranuclear and diffuse intracytoplasmic immunostaining of hepatocytes, predominantly in the periportal areas. In some cases there was also positive staining of macrophages in the lungs (4 cases), spleen (4 cases) and in lymph nodes (1 case). Positive granular staining in the renal mesangial cells of the glomeruli was observed in 1 case. No positive staining was observed in the 3 negative cases. In contrast to other reports (4), crossreactivity of these antigens to Porcine Parvovirus (PPV) could not be confirmed. Furthermore the RHD virus (RHDV) seems not to crossreact with Feline and Bovine Parvoviruses.