Epidemiology of viral haemorrhagic disease and myxomatosis in a free-living population of wild rabbits

From January 1993 to June 1996, the epidemiology of myxomatosis and viral haemorrhagic disease (VHD) was studied in a free-living population of wild rabbits (Oryctolagus cuniculus) in Spain by means of serological surveys and radiotracking. Myxomatosis was endemic and associated with the breeding period. Its serological pattern was characterised by a 100 per cent prevalence of antibodies in adult rabbits and a rapid increase in antibodies in young rabbits in their first year. No mortality from myxomatosis was detected in adults, and mortality in young rabbits could not be estimated because of interference by predators and scavengers and the deaths of many radiotagged rabbits inside their burrows. VHD was also an endemic disease associated with the breeding period. Adults had a higher prevalence of antibodies against VHD than young rabbits, reaching values of 80 to 90 per cent. During the study, there was an increase in rabbit numbers as a result of a decrease in mortality from predation which was associated with an increase in mortality due to VHD and in the prevalence of antibodies to VHD. Mortality from VHD was lower in rabbits with VHD antibodies than in seronegative rabbits, but some mortality from the disease was also detected in seropositive rabbits. The annual mean mortality rate due to VHD in adult rabbits was estimated to be 21.8 per cent.     

Adult rabbits acquire resistance to lethal calicivirus infection by adoptive transfer of sera from infected young rabbits

Calicivirus infection of adult rabbits induces the so-called rabbit haemorrhagic disease (RHD) that kills 90% or more of the infected animals; in contrast, young rabbits (up to 8-week-old animals) are resistant to the same infectious agent. We report that calicivirus inoculation of young rabbits induced moderate titres of antiviral antibodies. When these rabbits reached adulthood, a second calicivirus inoculation resulted in resistance to RHD and boosting of antibody titres in half of the rabbits. Adoptive transfer of sera from calicivirus-infected young rabbits to na?ve adult rabbits conferred resistance to RHD. We conclude that calicivirus infection of young rabbits induces specific anti-calicivirus antibodies that will protect them from RHD when they reach adulthood.     

Seroprevalence of antibodies against gram-negative core antigens in rabbits, using an Escherichia coli J5 antigen-capture enzyme-linked immunosorbent assay

OBJECTIVE: To determine the seroprevalence of antibodies to gram-negative core antigens (GNCA) in specific-pathogen-free (SPF) rabbits (ie, free of Pasteurella multocida) and rabbits of undefined bacterial status (conventional). SAMPLE POPULATION: Serum samples were obtained from 7 groups of rabbits. The SPF rabbits comprised 2 adult groups and 1 immature group, whereas the 4 groups of conventional rabbits were all adults. PROCEDURE: A seroprevalence survey was conducted on rabbit sera for antibodies against GNCA, using an Escherichia coli J5 antigen-capture ELISA. RESULTS: Collective geometric mean titer (GMT) of adult rabbits was 1:6,463. The GMT of each of the 6 groups of adult rabbits was 1:956, 1:1,133, 1:4,525, 1:5,338, 1:7,669, and 1:25,600. Titers of populations differed significantly. CONCLUSION: Data analysis revealed there were anti-GNCA antibodies in rabbits. Similar to other species, the prevalence of IgM and IgG anti-GNCA antibodies increased with age. The IgG response was more marked than the IgM response. The SPF rabbits had lower IgG anti-GNCA titers than conventional rabbits, indicating possible cross-reactive epitopes between P multocida and Enterobacteriaceae. Rabbits with the highest anti-GNCA titers were those used in polyclonal antibody production, possibly stemming from endotoxin contamination of antigen or adjuvant. CLINICAL RELEVANCE: The possible cross-reactive antibodies directed at homologous wall components of Pasteurellaceae and Enterobacteriaceae could prove to be a possible heterotypic vaccination strategy for the protection of rabbits against pasteurellosis. Investigators should determine whether antigen impurity (endotoxin contamination) influences epitope focus during polyclonal antibody production and whether it affects sera variability among rabbits.     

Outcome of equids with clinical signs of West Nile virus infection and factors associated with death

OBJECTIVE: To determine outcome of equids in the western United States with clinical signs of West Nile virus (WNV) infection and identify factors associated with risk of death in infected equids. DESIGN: Cross-sectional study. ANIMALS: 484 equids in Nebraska and Colorado. PROCEDURE: Owners of 484 equids with laboratory-confirmed West Nile virus infection in Nebraska and Colorado were contacted by telephone, and a questionnaire was used to obtain information on signalment, management, clinical signs, date of disease onset, duration of disease, WNV vaccination status, and health status at the time of the interview. RESULTS: 137 of 482 (28.4%) animals died or were euthanatized. Ataxia, lethargy, muscle fasciculations, and weakness were the most common clinical signs of disease. Animals > or = 3 years old were more likely to die than were animals < or = 2 years old. Unvaccinated equids were twice as likely to die as were animals that had been vaccinated at least once prior to the onset of disease. Animals that were recumbent and unable to rise were 78 times as likely to die as were animals that never lost the ability to rise. Females were 2.9 times as likely to die as males. Two hundred seventy-one of 339 (79.9%) animals that survived recovered fully; mean duration of disease for these animals was 22.3 days. CONCLUSIONS AND CLINICAL RELEVANCE: Among equids with WNV infection, age, vaccination status, an inability to rise, and sex were associated with the risk of death.     

Efficacy of oral vaccination in the final stage of fox rabies elimination in Switzerland

Subsequent to rabies vaccination campaigns, two well-established methods for the determination of the proportion of vaccinated foxes--the detection of tetracycline (TC) in bones and the detection of virus neutralizing antibodies (VNA) in thoracic fluids--were used and compared. Special emphasis was given to the effect of a new method of bait distribution at the den, which is primarily targeted at young foxes. The overall proportion of vaccinated animals estimated by TC was 60% as compared to 50% by VNA. In young foxes overall, significantly lower proportions of vaccinated animals (58% by TC and 40% by VNA) than in adult foxes (75 and 59%) were estimated with both methods. Low proportions of vaccinated young animals were found after spring (39 and 18%), but also after autumn vaccination (56 and 35%). In contrast, after den vaccination the level of vaccination of young foxes reached that of adult foxes. The theoretical implication of the successful elimination of fox rabies in Switzerland in spite of a relatively low overall proportion of VNA-positive animals is discussed.     

The influence of feedlot management on an outbreak of bovine respiratory disease

Epidemic curves, odds ratios and chisquare were used to investigate an epidemic of respiratory disease in a pen of feedlot cattle. The cattle were divided into four groups by an eartag number received at processing. Data for each group were collected from feedlot records describing purchase, transportation and processing histories, daily feeding methods, daily pen movements, daily diagnoses, treatment and mortality rates. These data were used to describe the effects of market origin and feeding management on the levels and distribution of respiratory disease.

The three groups of cattle purchased from auction markets and started on high levels of grain in their rations were determined to be 6.3 times (P<0.0005) more likely to be treated for any disease, 4.9 times (P<0.0005) more likely to be treated for respiratory disease, 12.7 times (P<0.0025) more likely to die, and 6.7 times (P<0.0471) more likely to die with respiratory disease than the group made up primarily of farm-assembled heifers and started on a 10% grain ration with time for adjustment to grain.

     

Leukocyte-hepatocyte interaction in calicivirus infection: differences between rabbits that are resistant or susceptible to rabbit haemorrhagic disease (RHD)

Calicivirus infection is lethal for adult rabbits, whereas young rabbits (less than 8-weeks-old) are resistant to the same infectious agent. The virus replicates in the liver and causes a fulminant hepatitis in adult rabbits leading to rabbit haemorrhagic disease (RHD); this is in contrast with the mild and transient hepatitis observed in infected young rabbits. We have used electron microscopy to compare liver leukocyte infiltrates between young (resistant) and adult (susceptible) rabbits, 36-48 h after inoculation of the animals with caliciviruses. In adult rabbits, liver infiltrates were made up mostly of heterophils, and they were located near hepatocytes showing severe cellular damage. In contrast, liver leukocyte infiltrates of RHD-resistant young rabbits were dominated by lymphocytes that depicted membrane contacts with the cell surface of undamaged hepatocytes. We conclude that: (i) the cellular inflammatory response of the liver to calicivirus infection is different in rabbits that are susceptible (adult) or resistant (young) to RHD; (ii) leukocyte infiltration of the adult liver by heterophils is probably directed at the removal of dead hepatocytes, whereas the liver lymphocytic infiltration of young rabbits suggests the expression of viral antigens on the surface of liver cells of the RHD-resistant animals.     

Intranasal vaccination of rabbits with Pasteurella multocida A:3 outer membranes that express iron-regulated proteins

OBJECTIVE: To determine efficacy of intranasal vaccination of rabbits with Pasteurella multocida A:3 outer membrane proteins (OMP) expressing iron-regulated OMP (IROMP) in conferring protection against experimental challenge exposure. ANIMALS: 52 male New Zealand White rabbits. PROCEDURE: Rabbits were vaccinated intranasally on days 0, 7, and 14; some vaccines included cholera toxin (CT) as an adjuvant. Concentrations of intranasal IgA and serum IgG antibodies against P multocida OMP were determined. In experiment A, rabbits were vaccinated with either phospate-buffered saline solution (PBSS), PBSS-CT, OMP-CT, or IROMP-CT, challenge-exposed intranasally on day 16, and euthanatized and necropsied on day 28. Rabbits were also vaccinated with OMP or IROMP without CT and were not challenge-exposed. In experiment B, rabbits were vaccinated with PBSS, PBSS-CT, IROMP, or IROMP-CT. On day 17, rabbits were challenge-exposed intranasally. Nasal bacteria and antibodies were determined on day 24. RESULTS: In experiment A, OMP-CT vaccination stimulated mucosal and systemic antibody responses to the bacterium and enhanced resistance against challenge exposure. Intranasal bacterial counts were not significantly reduced. Vaccination with IROMP-CT stimulated mucosal and systemic antibodies, enhanced resistance to challenge exposure, and significantly reduced nasal bacterial counts. In experiment B, natural infection was detected in several rabbits at challenge exposure; however, IROMP-CT-vaccinated rabbits had significantly higher serum and nasal antibody responses, compared with other rabbits IROMP-CT-vaccinated rabbits had significantly lower nasal bacterial counts compared to control rabbits. CONCLUSIONS AND CLINICAL RELEVANCE: Intranasal vaccination of rabbits with P multocida outer membranes containing IROMP and CT stimulated immunity against experimental pneumonic pasteurellosis.     

Ingestion capacity of PMN cells in peripheral blood of rabbits experimentally infected with VHD (viral haemorrhagic disease) virus strains originating from various biotopes

The study presents results of investigations on ingestion capacity in rabbits experimentally infected with two French strains (Fr-1 and Fr-2) and four Polish strains (KGM, SGM, MAL, Kr-1) of VHD virus. The results proved that the strains studied exhibited distinct reactivity even if the VHD virus is thought to be uniform in the biological respect.     

Liver disease in young rabbits infected by calicivirus through nasal and oral routes

Calicivirus infection causes rabbit haemorrhagic disease (RHD) that kills more than 90% of adult animals, whereas young rabbits are naturally resistant to this viral disease. It has been proposed that the different response of adult and young rabbits to calicivirus infection is due to absence of viral receptors in respiratory and digestive systems of young animals. We have searched for liver disease in 4-week-old rabbits inoculated with a calicivirus suspension by intranasal and oral routes. These young rabbits showed cell damage and mononuclear infiltration of the liver. The hepatic lesions were associated with mild to moderate increase in circulating transaminases. We conclude that the previously reported reduction of viral receptors in the epithelium of respiratory and digestive systems of young rabbits does not inhibit calicivirus from inducing liver disease in these hosts.     

Inhibition of priming for bovine respiratory syncytial virus-specific protective immune responses following parenteral vaccination of passively immune calves

The effect of maternal antibodies (MatAb) on immunological priming by neonatal parenteral vaccination for bovine respiratory syncytial virus (BRSV) was addressed for the first time in experimental infection in 34 Holstein calves. Both vaccinated and control calves developed moderate to severe respiratory disease characteristic of acute BRSV infection. There were no differences in clinical signs, BRSV shed, arterial oxygen concentrations, or mortality between vaccinated and control calves after BRSV challenge approximately 11 wk after vaccination. There were no anamnestic antibody or cytokine responses in the vaccinates after challenge. Lung lesions were extensive in both groups, and although there was a statistically significant (P = 0.05) difference between groups, this difference was considered not biologically significant. These data indicate that stimulation of protective immune responses was inhibited by maternal antibodies when a combination modified-live BRSV vaccine was administered parenterally to young passively immune calves. Alternate routes of administration or different vaccine formulations should be used to successfully immunize young calves with good passive antibody transfer.     

Liver disease in young rabbits infected by calicivirus through nasal and oral routes

Calicivirus infection causes rabbit haemorrhagic disease (RHD) that kills more than 90% of adult animals, whereas young rabbits are naturally resistant to this viral disease. It has been proposed that the different response of adult and young rabbits to calicivirus infection is due to absence of viral receptors in respiratory and digestive systems of young animals. We have searched for liver disease in 4-week-old rabbits inoculated with a calicivirus suspension by intranasal and oral routes. These young rabbits showed cell damage and mononuclear infiltration of the liver. The hepatic lesions were associated with mild to moderate increase in circulating transaminases. We conclude that the previously reported reduction of viral receptors in the epithelium of respiratory and digestive systems of young rabbits does not inhibit calicivirus from inducing liver disease in these hosts.     

“黄白散”对兔接种RHD疫苗后免疫功能的影响

在注射兔病毒性出血症 (RHD)疫苗后 ,30只兔随机分为 2组 ,试验组兔饲喂含 1 5 %黄白散的饲料。从接种到第 1 80天 ,每组取兔 8只 ,每隔 1 5d采血 1次 ,检测血凝抑制 (HI)抗体 ;在接种前 1天及接种后 5、 1 0、 1 7、 2 4、 31、 41、 5 1d ,每组取兔 1 0只 ,采血检测ANAE+淋巴细胞百分率 ;在 1 80、 2 4 0、 30 0d时 ,每组分别取兔 5只做攻毒保护试验。结果表明 :2组兔HI抗体峰值分别为 7 3log2和 9 8log2 ,差异极显著 (P <0 0 1 ) ;试验组疫苗保护期可延长 60d ;试验组ANAE+率从第 1 0天至 5 1天与对照组比较 ,差异极显著(P<0 0 1 )。所以黄白散可增强兔接种RHD疫苗后的特异性免疫功能 ,增强RHD疫苗的免疫效果。     

Regulatory T cells are decreased in acute RHDV lethal infection of adult rabbits

Rabbit hemorrhagic disease virus (RHDV) is the etiologic agent of rabbit hemorrhagic disease (RHD), an acute lethal infection that kills 90% of adult rabbits due to severe acute liver inflammation. Interestingly, young rabbits are naturally resistant to RHDV infection. Here, we have compared naturally occurring CD4(+)Foxp3(+) regulatory T cells (Tregs) between young and adult rabbits after infection by RHDV. The number and frequency of Tregs was decreased in the spleen of adult rabbits 24h after the RHDV infection; this was in contrast with the unchanged number and frequency of splenic Tregs found in young rabbits after the same infection. Also, serum levels of IL-10 and TGF-β were enhanced in the infected adult rabbits whereas no alteration was observed in infected young rabbits. However, this increase is accompanied by a burst of pro-inflammatory cytokines, but seems not able to prevent the death of the animals with severe acute liver inflammation in few days after infection. Since Tregs downregulate inflammation, we conclude that their decrease may contribute to the natural susceptibility of adult rabbits to RHDV infection.     

Serologic and bacteriologic test results after adult vaccination with strain 19 in three dairy herds infected with brucellosis.

Milk culture data and serologic test results were evaluated after adult vaccination with Brucella abortus strain 19 in cattle of 3 large California dairy herds infected with brucellosis. Strain-19 organisms were isolated by culture of milk from 1.9% of the vaccinated cows. Isolation of field strain of B abortus varied directly with magnitude of complement-fixation (CF) and rivanol titers. At time of milk culture, 74% of cows from which field strain was isolated had CF titer greater than or equal to 160, compared with 58% of cows from which strain 19 was isolated. Cows with CF titer greater than or equal to 160 at 2 months or greater than or equal to 80 to 4 months after adult vaccination were more likely to be correctly classified as reactors (on the basis of subsequent milk culture results and/or persistently high serologic titer) than were cows with lower CF titer at these times. Cows from which B abortus strain 19 was isolated from milk were more likely to maintain persistent serologic titer than were cows from which neither strain of B abortus was isolated.     

Immunosuppression abrogates resistance of young rabbits to Rabbit Haemorrhagic Disease (RHD)

Rabbit Haemorrhagic Disease (RHD) is caused by a calicivirus (RHDV) that kills 90% of infected adult European rabbits within 3 days. Remarkably, young rabbits are resistant to RHD. We induced immunosuppression in young rabbits by treatment with methylprednisolone acetate (MPA) and challenged the animals with RHDV by intramuscular injection. All of these young rabbits died within 3 days of infection due to fulminant hepatitis, presenting a large number of RHDV-positive dead or apoptotic hepatocytes, and a significant seric increase in cytokines, features that are similar to those of naïve adult rabbits infected by RHDV. We conclude that MPA-induced immunosuppression abrogates the resistance of young rabbits to RHD, indicating that there are differences in the innate immune system between young and adult rabbits that contribute to their distinct resistance/susceptibility to RHDV infection.     

Preliminary testing in turkeys of the safety and efficacy of a putative haemorrhagic enteritis virus vaccine

Haemorrhagic enteritis virus (HEV) causes clinical haemorrhagic enteritis in young poults and/or subclinical immunosuppression which is often associated with colibacillosis. This disease is controlled with live vaccines worldwide, however, importation of HEV vaccines or cells that support HEV propagation are not permitted in Australia. A major experiment in isolators was conducted to test the safety and efficacy of a putative HEV vaccine. The study had a factorial design with four factors namely vaccination age (28 and 42 days of age), vaccine dose (0, 10⁵, 10⁶, 10⁷ genomic copies of HEV vaccine), challenge with HEV (yes, no) and vaccination‐challenge interval (7, 21 or 42 days). A total of 315 poults were used providing 6‐8 birds per treatment combination. Turkey growth rate, mortality, pathological findings, anti‐HEV antibodies and viral load were examined. Vaccination lead to significant increases in anti HEV antibody over the following 2‐4 weeks. Overall, vaccination with 10⁶ and 10⁷ was protective against increase in relative splenic weight and splenic viral load in challenged birds. Clinical haemorrhagic enteritis was not induced by any treatment but there was an increased incidence of airsacculitis in groups receiving either HEV vaccine or challenge virus compared to the negative control birds (25.8‐29.3% vs. 9.4%, P < 0.05). Growth rate, mortality and relative bursal weight were unaffected by vaccination. This laboratory level study indicates that the putative vaccine is safe and likely to be efficacious, but may cause elevated levels of airsacculitis. These findings require confirmation in larger scale field trials.     

Growth Performance and Reproductive Traits at First Parity of New Zealand White Female Rabbits as Affected by Heat Stress and Its Alleviation under Egyptian Conditions

Exposing growing and adult New Zealand White (NZW) female rabbits to severe heat stress (temperature–humidity index = 28.9) during summer adversely affected their growth and reproductive traits. The traits that declined significantly (p<0.01) were the live body weight, daily weight gain and feed intake of growing rabbits, and the litter size and litter weight at weaning (p<0.05) and the pre-weaning weight gain of pups (p<0.01) for adult females. The conception rate declined considerably with heat stress. The declines in the values of the digestibility coefficients due to heat stress were 7.9% (p<0.05) for dry matter (DM), 8.1% (p<0.05) for crude protein (CP) and 1.0% for crude fibre (CF). The traits that increased significantly (p<0.01) due to heat stress were water intake, water/feed ratio and rectal temperature in growing rabbits and pre-weaning mortality for adult females.Alleviation of heat stress in the growing and adult female NZW rabbits was more efficient with drinking cool water (10–15°C; between 10:00 and 17:00) than with supplementation with palm oil (as a source of energy) or natural clay (as a natural enhancer to growth and milk production). Supplying the animals with cool drinking water gave the highest body weight and weight gain, conception rate, litter size and weight and digestibility coefficients for DM and CP and the lowest rectal temperature, respiration rate and pre-weaning mortality.The loss in rabbit production pertaining to heat stress estimated from the percentages of decline in conception rate×pre-weaning mortality×litter weight at weaning was 73.0%. The provision of cool water restored 11/12 of heat loss.     

Evidence of peste des petits ruminants virus (PPRV) infection in Sindh Ibex (<Emphasis Type="Italic">Capra aegagrus blythi</Emphasis>) in Pakistan as confirmed by detection of antigen and antibody

An outbreak resulting in mortality in Sindh Ibex (Capra aegagrus blythi) was investigated. There was a history of about 36 deaths (both young and adult) during the period of 1 month. Disease appeared in a generalized form, affecting the respiratory and digestive systems. Major lesions were respiratory distress, pustules on and in the mouth, ocular–nasal discharges, and severe diarrhea. The most significant lesion was the oculonasal discharges and diarrhea. Deaths were mainly due to blindness, anorexia, diarrhea, and respiratory arrest. Both adult (mortality = 21) and young (mortality = 15) animals were affected with the disease. Peste des petits ruminants virus (PPRV) antigen was detected in the spleen, lung, lymph node, and swab samples by immunocapture enzyme-linked immunosorbent assay. Spleen and lung samples were also tested and found positive for the presence of F-gene of PPRV by polymerase chain reaction. Thirteen of 20 serum samples from nearby sheep and goats were found positive for antibodies to PPRV. The disease threatened the huge population of ibex in the wild life park, which was spread over a large area, but vaccination of the domestic population of sheep and goats in the surrounding villages appeared to control the disease.     

Oral immunisation of swine with a classical swine fever vaccine (Chinese strain) and transmission studies in rabbits and sheep

Seven experiments including a total of 47 pigs, 11 wild boars, 26 rabbits, 10 hares and 16 sheep were carried out to assess the efficacy, safety and transmission of the Chinese vaccine strain of the classical swine fever virus (CSFV) administrated by the oral route. Within 3 weeks after oral vaccination, a clear seroconversion occurred in the pigs. Six weeks after vaccination, vaccinated pigs were fully protected against a virulent challenge. The C-strain was not isolated from tonsils, spleen, lymph nodes, thymus, saliva, urine and faeces of pigs within 4 days after oral vaccination. In one experiment, susceptible pigs were placed in direct contact with vaccinated pigs. None of these contact-exposed pigs became serologically positive for CSFV antibodies. It is concluded that the C-strain induces protection in pigs when administrated by the oral route and is not shed by vaccinated pigs. Serum anti-CSFV antibodies developed in seven out of eight wild boars vaccinated by the oral route. No vaccine virus was detected in the spleen and tonsils of these animals. The results in wild boar were in accordance with those obtained in domestic pigs. Sheep did not show any clinical signs after oral vaccination while rabbits had moderate hyperthermia and growth retardation. No clinical response to oral immunisation in hares was detected. At the end of the experiment, no sheep had detectable serum antibodies against CSFV, whereas a few vaccinated rabbits and hares became seropositive. None of the contact-exposed rabbits and hares seroconverted. These data indicate that the C-strain is safe for sheep and as expected, moderately or not pathogenic for rabbits and hares. These efficacy and safety studies on oral vaccination with the C-strain under experimental conditions provide essential information for further studies in wild boars under experimental and field conditions, including assays with baits to control a CSF epidemic.