Immunological relationships between malignant catarrhal fever virus (alcelaphine herpesvirus 1) and bovine cytomegalovirus (bovine herpesvirus 3)

Strains of malignant catarrhal fever virus (alcelaphine herpesvirus 1 (AHV-1)) and bovine cytomegalovirus (bovine herpesvirus 3 (BHV-3)) were compared for serological relatedness by cross-titration in an indirect immunofluorescent (IIF) antibody assay. There was definite cross-reactivity between these 2 viruses, with heterologous sera staining intracellular and membrane antigens of infected cells. Heterologous antibody titres were approximately 50-fold lower than homologous titres and could be removed by absorption with either homologous or heterologous virus-infected cells, but not with uninfected cells. Regression analyses of IIF antibody titres to AHV-1 and BHV-3 virus in 3 groups of wild ungulate sera also indicated a serological relationship between these herpesviruses. In a cross-immunity trial, 2 of 3 cattle immunized with a BHV-3 virus and 2 of 3 cattle immunized with avirulent AHV-1 resisted challenge with virulent AHV-1-infected blood which killed 3 unimmunized controls. These results are discussed particularly with respect to the involvement of BHV-3 in malignant catarrhal fever.     

Antibody to alcelaphine herpesvirus-1 (AHV-1) in hamsters experimentally infected with AHV-1 and the 'sheep-associated' agent of malignant catarrhal fever

Malignant catarrhal fever was induced in four groups of hamsters by the inoculation of cells infected with either the C/500 isolate of alcelaphine herpes-virus-1 (AHV-1) or the sheep-associated agent derived from cattle, red deer or Père David's deer. Using an indirect immunofluorescence assay, antibody to AHV-1 was detected in sera of clinically affected animals of all four groups. The reaction of sera from hamsters affected with malignant catarrhal fever induced by AHV-1 caused diffuse cytoplasmic staining while that from sera of hamsters with the sheep-associated form of the disease stained particulate nuclear antigens. Tests employing three other bovid herpesviruses were negative and no reaction was found with sera from normal hamsters. These studies provide convincing evidence that a virus antigenically related to AHV-1 is the cause of sheep-associated malignant catarrhal fever and that the same virus probably causes this form of the disease in both cattle and deer.     

Development of an enzyme-linked immunosorbent assay for the detection of antibodies against malignant catarrhal fever viruses in cattle serum

Malignant catarrhal fever (MCF) is a sporadic but fatal lymphoproliferative viral disease of cattle, deer and other ruminants. The causative agents are highly-cell-associated herpesviruses of the subfamily gammaherpesvirinae. In this study, an ELISA (WC11-ELISA) was developed to detect antibody to malignant catarrhal fever virus (MCFV) in cattle serum and compared to the commercially produced competitive-inhibition ELISA (CI-ELISA). Crude lysate antigen from alcelaphine herpesvirus-1 strain WC11 was bound to 96-well microplates and used to capture antibodies to MCFV. Dilutions of test sera were added to wells containing bound MCF antigen and control wells containing uninfected cell lysates. A horseradish peroxidase-labelled rabbit-anti-bovine IgG conjugate detected antibodies to MCF, and the results were expressed as absorbance readings at 450 nm. Samples were selected blind from cattle sera which had been sent to the laboratory for diagnostic testing for MCFV antibodies and were tested in both the WC11-ELISA and the CI-ELISA. Good agreement between the WC11-ELISA and CI-ELISA test (k=0.86, n=95) results was found.     

Studies concerning etiology of sheep-associated malignant catarrhal fever in Europe.

The etiological agent of sheep-associated malignant catarrhal fever (MCF) in Europe has not been isolated directly from sheep. The occurrence of antibodies against the African bovine herpesvirus (BHV-3, WC 11) in cattle and sheep was examined using recently modified indirect immunofluorescence test (IIFT) and neutralization test (NT) methods. Studies revealed that sheep and cattle sera in Europe were free from neutralizing antibodies of BHV-3. The IIFT could not establish the presence of antibodies to African BHV-3 in cattle but revealed that about 22.4% of sheep sera reacted to it. Apart from the well known ovine herpesvirus (BHV-5), occurrence of another herpesvirus in Europe had been expected. This virus is not identical with the WC 11 strain, but it is in antigenic relationship to it. We had for the first time substantial serological evidence to the effect that sheep-associated MCF in Europe is a herpesvirus related to the African strain.     

Concurrent malignant catarrhal fever and bovine virus diarrhoea virus infection in a dairy herd

An account is given of an outbreak of malignant catarrhal fever which occurred in a 98-cow dairy herd. Ten animals died or were slaughtered and the disease was confirmed by clinical and histological examination. Serological tests for malignant catarrhal fever virus were positive in three of four animals. The diagnosis of malignant catarrhal fever was complicated by the presence of bovine virus diarrhoea virus infection in three of the early cases. The initial cases of malignant catarrhal fever occurred in a group of nine-month-old calves which were housed in an old milking parlour with 19 pedigree Suffolk ewes at lambing time. Later cases occurred in two adult cows and in two heifers. Investigations of the remainder of the herd for evidence of bovine virus diarrhoea virus did not reveal the presence of any persistently infected cattle. Serological examinations for antibody to malignant catarrhal fever and bovine virus diarrhoea virus were carried out on the 19 Suffolk ewes. Six of them had neutralising antibody titres to malignant catarrhal fever virus and three were positive in the indirect immunofluorescence test. The possible roles of bovine virus     

Outbreak of malignant catarrhal fever in Welsh Black cattle in Carmarthenshire

An outbreak of malignant catarrhal fever (MCF) resulted in the deaths of 12 cattle in a herd of 77 animals during seven weeks in 1999; in addition, one cow developed a milder disease which was confirmed as MCF by PCR for ovine herpesvirus 2 DNA and an immunofluorescent antibody test for antibodies to the virus, but recovered. Further PCR and serological testing revealed the infection in three other animals, none of which developed clinical disease. Hypocuprosis and the possibility of a genetic predisposition were identified as factors which may have contributed to the outbreak.     

Antibody response in cattle and rabbits to early antigens of malignant catarrhal fever virus in cultured cells.

Cytosine arabinoside (Ara-C) inhibited the production of late antigens and of infectious virus in monolayers of bovine kidney cells infected with the high-passage, WC-11 strain of malignant catarrhal fever virus. Early antigens were not affected. Using hyperimmune and acute-phase sera from cattle and rabbits in indirect immunofluorescence tests, it was shown that Ara-C treated cultures contained two early antigens; one was diffuse and distributed throughout the cells, the other was particulate and intranuclear. Antibody to early antigens developed later and attained lower titres in infected animals, especially rabbits; only hyperimmune sera reacted with the diffuse early antigen.     

Antigens and antibodies of malignant catarrhal fever herpesvirus detected by immunodiffusion and counter-immunoelectrophoresis

Using hyperimmune rabbit and cattle sera, immunodiffusion (ID) and counter-immunoelectrophoresis (CIEP) tests detected three or four and two or three malignant catarrhal fever (MCF) virus antigens, respectively, in infected cells. The ID test detected precipitating antibodies to MCF virus in $\text{3}\text{9}$ experimentally infected rabbits, $\text{0}\text{14}$ experimentally infected cattle, $\text{1}\text{13}$ naturally infected cattle, $\text{62}\text{176}$ wildebeest and $\text{3}\text{20}$ hartebeest. The CIEP test detected specific antibodies in $\text{3}\text{9}$ rabbit sera, but non-specific reactions prevented its use with bovine sera.The CIEP test was 2 to 4 times more sensitive than ID for detecting antibodies to MCF virus, but both tests were less sensitive than indirect immunofluorescence.The ID test demonstrated an antigenic relationship between wildebeest and hartebeest strains of MCF virus. Neither ID nor CIEP detected MCFV antigens in tissues infected with MCF virus.     

The effect of antibody and complement on the expression of herpesvirus of bovine malignant catarrhal fever in cultured rabbit lymphocytes

Using indirect immunofluorescence with a hyperimmune calf serum, a virus-induced antigen was demonstrated on the surface of lymphocytes expressing intracellular malignant catarrhal fever virus antigens. Antibody to the antigen was also detected in terminal sera of both cattle and rabbits. Antisera did not restrict virus expression in explanted lymph nodes unless they were supplemented with two to four units of lytic complement per ml culture. While human, bovine and guinea pig complements caused immune lysis of infected lymphocytes, rabbit complement was ineffective. The relevance of the findings in the pathogenesis of the lymphoid proliferation caused by MCFV is discussed.     

Role of wildebeest fetal membranes and fluids in the transmission of malignant catarrhal fever virus

Malignant catarrhal fever virus was not isolated from samples of fetal membranes or fluid collected from 93 calving wildebeest (Connochaetes taurinus) in Kenya Maasailand. Cell-free strains of malignant catarrhal fever virus were very rapidly inactivated when exposed to the sun under field conditions, at least 3.0 log10 units/25 microliter being lost per hour at midday. It is suggested that wildebeest fetal membranes and fluids act as visual markers for areas of pasture which are particularly heavily contaminated with malignant catarrhal fever virus in oculonasal secretions of wildebeest calves. It is possible that starting to graze cattle one to two hours later each morning may be a useful measure for helping to protect cattle from malignant catarrhal fever in areas where they are forced to share pastures with calving wildebeest.     

Antibodies in carrier wildebeest to the lymphoproliferative herpesvirus of malignant catarrhal fever

Six types of antibody to malignant catarrhal fever virus (MCFV) were measured in 132 sera collected from Wildebeest in Kenya Masailand. The titre of all types of antibody declined slowly with increasing age of the wildebeest. A significantly greater proportion of wildebeest calves had higher titres of antibodies to MCFV early antigens, IgM antibodies to MCFV late antigens and complement-fixing antibodies, than did older animals. One seronegative calf, reared in isolation without colostrum, became seropositive 4 1/2 weeks after birth but did not show any clinical signs indicative of MCFV infection. Similarities between MCFV infection of wildebeest calves and other inapparent infections with lymphoproliferative herpesviruses are discussed.     

Occurrence of ovine herpesvirus type-2 infection in sheep and cattle in Samsun Province, Turkey

July 2004, a cow with clinical signs of ovine herpesvirus type-2 infection which is known as sheep associated malignant catarrhal fever (SA-MCF) was reported in Samsun Province in Turkey. Blood samples were collected from the suspected cow, 10 sheep housed with it, and from 150 healthy sheep and 29 healthy cattle randomly selected from different places in Samsun Province. Nested polymerase chain reaction (n-PCR) was used to detect ovine herpesvirus type-2 (OvHV-2) DNA in the suspected cow and competitive- ELISA (c-ELISA) kits were used to detect antibodies against OvHV-2. The suspected cow was found to be n-PCR positive and c-ELISA negative. The serological results were as follows: All 10 (100%) of sheep housed with the suspected cow and 18 of 29 (62%) of the randomly selected cattle were found seropositive. All 150 randomly selected healthy sheep were seronegative. The overall percentage of seropositivity was 14.7% (28/190). OvHV-2 DNA was detected in the peripheral blood leucocyte (PBL) samples of the cow and of the 10 sheep housed with the suspected cow.     

Malignant catarrhal fever. I. Response of American cattle to malignant catarrhal virus isolated in Kenya.

Fifty-three American cattle were inoculated with malignant catarrhal fever virus isolated from a wildebeest in Kenya. Three animals showed the mild form of the disease and recovered, and 47 showed the severe form of the disease. The other three did not react. Of the 47 cattle, 28 died, 16 were killed for the collection of specimens and three recovered. The incubation period for the 47 cattle ranged from 16 to 29 days and the course of the fatal disease for 28 cattle averaged three to 23 days. Virus titration of specimens from nine infected steers yielded a mean titer of 10(4)/TCID50 per gm for lymph nodes, 10(3) TCID50 per mL for buffy coats and 10(2.3) TCID50 per gm for spleens. Smaller amounts of virus were found in the liver, kidneys, adrenals and thyroids. Malignant catarrhal fever virus was also found in nasal secretions and saliva of viremic cattle. Viral infectivity was shown in bovine buffy coat cells stored at 4 degrees C for two days but was immediately destroyed upon freezing even when glycerine or dimethylsulfoxide was added. Viral particles were not found in infected animal tissues by electron microscopy. The disease was successfully transmitted in steers by intratracheal intubation and by aerosol inhalation but not by contact.     

Detection of African malignant catarrhal fever virus antigens in cell cultures by immunofluorescence

A fluorescent antibody (FA) test for antigens of African bovine wildebeest-derived malignant catarrhal fever virus was developed. Serum from one of the few survivors of the experimental disease in steers was used to prepare the conjugate. Both a virulent and an attenuated strain of malignant catarrhal fever virus were used to infect bovine thyroid cell cultures. Cells infected with both strains were readily detected by FA staining as early as 24 h post infection, whereas cytopathic effect could be observed by bright-field microscopy only after days 5 or 6 post infection. Controls consisting of normal bovine thyroid cells or infected cells treated with conjugated normal globulins did not show autofluorescence. The reaction was blocked by treatment of infected cells with homologous positive antisera but not by treatment with normal bovine serum or antisera to foot-and-mouth disease, rinderpest, bovine virus diarrhea, Ibaraki, infectious bovine rhinotracheitis, or bovine herpes mammilitis viruses. Treated with African malgnant catarrhal fever virus conjugate did not react.     

Malignant catarrhal fever virus specific secretory IgA in nasal secretions of wildebeest calves

Wildebeest IgA was isolated from nasal secretions and precolostrum. It was indentified by cross-reaction with anti-human and anti-bovine IgA sera.

Nasal secretions collected from wildebeest calves over 3 months old had malignant catarrhal fever virus neutralizing antibody activity. They also contained specific IgA to the virus as detected by indirect immunofluorescence. It is suggested that production of malignant catarrhal fever virus specific IgA in the nasal cavity, contributes to the elimination and cassation of the virus shed in the nasal secretions of wildebeest calves over 3 months. old.     

A simpler, more sensitive competitive inhibition enzyme-linked immunosorbent assay for detection of antibody to malignant catarrhal fever viruses.

An earlier competitive inhibition enzyme-linked immunosorbent assay (CI-ELISA) was developed for detection of specific antibody against malignant catarrhal fever (MCF) viruses (MCFV) in ruminants. In this study, the indirect CI-ELISA was improved by conjugating the monoclonal antibody 15-A directly with horseradish peroxidase and by developing a method of producing precoated, dried antigen plates. This new test is referred to as a direct CI-ELISA. The reformatted test yielded a significantly improved sensitivity, and the time required was reduced to about one-sixth of the previous time. Of 37 MCF cases in cattle that were confirmed by histopathology and polymerase chain reaction (PCR) assay, 37 (100%) were positive by the new test, whereas the indirect CI-ELISA detected only 23 (62%). The direct CI-ELISA detected antibody to MCFV in 100% of 48 sheep that had been defined as infected with ovine herpesvirus 2 (OvHV-2) by PCR, whereas the indirect CI-ELISA detected only 41 (85%). Comparison of antibody titers measured by the 2 assays for sera collected from OvHV-2-infected sheep and from cattle, bison, and deer with clinical sheep-associated MCF revealed that the direct CI-ELISA offered a 4-fold increase in analytical sensitivity over the indirect format. The number of seropositive animals detected by the direct CI-ELISA among apparently normal cattle and bison was 2-3 times greater than the number detected by the indirect CI-ELISA, indicating that a significant percentage of normal cattle and bison are subclincally infected with MCFV.     

Attempts to immunize cattle against virulent African malignant catarrhal fever virus (alcelaphine herpesvirus-1) with a herpesvirus isolated from American cattle.

Two consecutive weekly inoculations with a herpesvirus isolated from sick cattle in America (707K), protected four out of four steers against a first challenge with virulent African malignant catarrhal fever virus (alcelaphine herpesvirus-1), strain C500. Three of these steers were still protected in a rechallenge carried out 9.5 months after the first challenge. One inoculation with this agent did not protect such steers, and repeated weekly inoculations had the risk of inducing a malignant catarrhal fever-like disease. In addition such repeated inoculation did not necessarily confer adequate protection, either in the first or the second challenge. There was no correlation between the development of virus neutralizing antibody and protection to challenge with the virulent virus. Endonuclease analysis of the genome of 707K virus, revealed differences between the agent and the avirulent cell-free form of the virulent African malignant catarrhal fever virus (WC11).     

Neutralising antibodies to wildebeest-derived malignant catarrhal fever virus in African wildlife

A total of 2,722 sera collected between 1963 and 1983, from 43 different species of wildlife in 11 African countries was examined for neutralising antibodies against the wildebeest-derived strain of malignant catarrhal fever (MCF) virus. Antibodies were demonstrated in 10 species of Bovidae which included eight species from the sub-family Hippotraginae and one species each from Bovinae and Antilopinae. Neutralising antibodies were also recorded in hippopotamus. It is suggested that the high prevalence of antibodies recorded in sera from waterbuck and reedbuck indicate infection with MCF. However, titres in other species may be due to antigenically related viruses.     

Immune complexes associated with infection of cattle by the herpesvirus of malignant catarrhal fever

Associated with the fatal lymphoid proliferation induced by the herpesvirus of African malignant catarrhal fever (MCFV) was the deposition of immune complexes comprising immunoglobulin, complement and conglutinin in the kidneys of cattle with terminal pyrexia due to infection with this virus. Such deposits were common in glomeruli and were also seen in renal arteries with and without lesions. Absence of lytic complement and a significant depletion of conglutinin in terminal sera confirmed the activation of complement. However, virus antigen was only rarely detected and virus-specific antibody could not be demonstrated in the deposits. These disparities and the importance of the observations are discussed.     

The etiology and epidemiology of malignant catarrh--a review

It is generally accepted that both, the wildebeest-derived malignant catarrhal fever (WD-MCF), and the circumstantially evidenced sheep-associated form of the disease (SA-MCF), may be explained as autoimmune disease of various ruminants, namely cattle and farmed deer. The disease follows infection with related herpesviruses being shed by the respective healthy carrier animals. This has convincingly be shown to apply for WD-MCF (Alcelaphine herpesvirus 1, AlcHV1). SA-MCF, however, remains to be controversial with both respects. In Switzerland, a serological study indicated that a herpesvirus(es) was highly prevalent among cattle and sheep, inducing antibody that cross-react with AlcHV1 and bovine herpesvirus 4 (BHV4). The latter is known as a largely innocuous agent. A relationship can be demonstrated between the presence of MCF in this country and concurrent serological reactions to both viruses. However similar results may be obtained with healthy animals. Healthy cattle and sheep from farms with or without incidences of MCF displayed the same antibody profiles. It is thus not possible to effectuate meaningful diagnostic tests for (SA-)MCF, nor to confirm any relationship between presumed carrier sheep and the appearance of MCF.