http://repub.eur.nl/res/aut/457/ List of Publications en http://repub.eur.nl/static-eur/img/logo.png http://repub.eur.nl http://repub.eur.nl/res/pub/3921/ 2003-07-01T00:00:00Z Phocid herpesvirus type 2 (PhHV-2), tentatively classified as a gammaherpesvirus, has been isolated from European and American harbour seals (Phoca vitulina). Here we describe the isolation and the molecular as well as biological characterisation of different PhHV-2 isolates from harbour seals and grey seals (Halichoerus grypus). Of 522 harbour seals and 231 grey seals that had been admitted to the seal research and rehabilitation centre in Pieterburen, The Netherlands, between 1992 and 2000, 38 and 18%, respectively, proved to have PhHV-2 neutralising antibodies. PhHV-2 was isolated from peripheral blood mononuclear cells (PBMCs) of 12 and 28% of these seropositive animals, respectively, and 26 and 56% of these cell samples, respectively, were positive by PCR analysis. Analysis of amino acid sequences of PCR products and of the growth characteristics of different PhHV-2 isolates indicated that harbour and grey seals are infected with distinct γ-herpesviruses, which however, may co-circulate between the two species. http://repub.eur.nl/res/pub/3919/ 2003-06-01T00:00:00Z To further characterize phocid herpesvirus type 1 (PhHV-1) at the molecular level, a cluster of genes comprising the complete unique short (Us) region of PhHV-1 has been cloned and sequenced. Within this region, ORFs were detected that code for the equivalent of the Us 2- protein of herpes simplex virus (HSV), a putative protein kinase, and for the glycoprotein equivalents gG, gD, gI and gE. In addition, two small ORFs downstream of gE, homologous to the Us 8·5 and Us 9 proteins of HSV were identified. Comparative analysis of the ORF encoding the gD equivalent of PhHV-1 identified the corresponding proteins of the alphaherpesviruses canine herpesvirus and, to lesser degree, feline herpesvirus as the closest relatives. http://repub.eur.nl/res/pub/8473/ 2003-01-01T00:00:00Z To further characterize phocid herpesvirus type 1 (PhHV-1) at the molecular level, a cluster of genes comprising the complete unique short (Us) region of PhHV-1 has been cloned and sequenced. Within this region, ORFs were detected that code for the equivalent of the Us 2- protein of herpes simplex virus (HSV), a putative protein kinase, and for the glycoprotein equivalents gG, gD, gI and gE. In addition, two small ORFs downstream of gE, homologous to the Us 8.5 and Us 9 proteins of HSV were identified. Comparative analysis of the ORF encoding the gD equivalent of PhHV-1 identified the corresponding proteins of the alphaherpesviruses canine herpesvirus and, to lesser degree, feline herpesvirus as the closest relatives. http://repub.eur.nl/res/pub/3830/ 2001-12-01T00:00:00Z Phocid herpesvirus type 1 (PhHV-1) causes significant morbidity and mortality among young and immunocompromised harbour seals. Therefore, the availability of an effective PhHV-1 vaccine would be of importance for orphanages and seal rehabilitation centres. Since possibilities to test PhHV-1 candidate vaccines in the target species are limited, a suitable animal model is needed. Given the close genetic and antigenic relationships between PhHV-1 and feline herpesvirus (FHV), the FHV cat system could be considered to test candidate PhHV-1 vaccines. Here we have tested a PhHV-1 based ISCOM vaccine for its protective efficacy against FHV infection in cats. To this end, three groups of cats were vaccinated thrice with ISCOM adjuvanted PhHV-1, FHV, and mock vaccines, respectively. One month after the last vaccination, all cats were challenged with a virulent FHV strain. All PhHV-1 and FHV vaccinated cats were protected from developing severe disease and excreted significantly less FHV than the mock vaccinated cats. http://repub.eur.nl/res/pub/18423/ 1997-10-29T00:00:00Z The Herpesviridae represent a family of diverse and complex viruses of vertebrates (Roizman et al., 1992; Roizman, 1996) and are believed to be of comparatively ancient origin (Karlin el al., 1994a; McGeoch et al., 1995). Many mammalian species are host to more than one herpesvirus species. In humans, e.g., an eighth distinct herpesvirus species has recently been discovered (Chang et al., 1994; Moore et aI., 1996). Disease associated with lytic herpesvirus infection in their natural hosts varies considerably from mild, superficial mucocutaneous lesions, acute respiratory disease, benign Iymphoproliferative disorders to fatal generalized infections and congenital malformations (Peterslund, 1991). Besides certain viral factors, immuno-(in)competence of the natural host constitutes a major pathogenic factor (Fawl & Roizman, 1994). Some herpesviruses also display transforming properties (e.g. Meinl et al., 1995; Thorley-Dawson el al., 1996) or act as co-factors in tumorigenesis (Nazarin, 1979; Khanna et al., 1995; Mesri et al., 1996). Herpesviruses characteristically establish life-long latenl infections in their hosts (Stevens, 1994). Periodically, latent (silent) virus can become reactivated leading to limited lytic replication and shedding of infectious particles. Latently infected hosts, therefore, represent the major herpesvirus reservoir. http://repub.eur.nl/res/pub/3592/ 1997-03-01T00:00:00Z Canine distemper morbillivirus (CDV) induces a multisystemic, often fatal disease in a wide and seemingly expanding host range among the Carnivora. Several genotypes of an otherwise monotypic virus species co-circulate in a geographically restricted pattern. Interspecies transmissions frequently occur, often leading to devastating epizootics in highly susceptible or immunologically naive populations. http://repub.eur.nl/res/pub/3591/ 1997-01-20T00:00:00Z A glycoprotein B (gB) gene homologue was identified in a 5.4-kb BamHl genomic fragment of the phocid herpesvirus type-1 (PhHV-1) which represents a widespread and important pathogen of pinnipeds. Sequence analysis revealed a gB-specific open-reading frame comprising 881 amino acids. Phylogenetic analysis gave evidence for a close evolutionary relationship between PhHV-1 and members of the Varicellovirus genus of the alpha-Herpesvirinae and canid herpesvirus in particular. In PhHV-1-infected Crandell feline kidney cells gB is expressed as a 113-kDa glycosylated molecule which is proteolytically cleaved into at least two fragments of 67 and 53-59 kDa apparently forming disulfide-linked heterodimers of 140 kDa. Cell surface expression of PhHV-1 gB was confirmed by FACS analysis. Thus, synthesis and processing of the gB protein of PhHV-1 follows a pattern also observed in other Varicelloviruses. Since the gB protein of herpesviruses, expressed in the baculovirus system, has been shown to be a suitable target for vaccine design, we used this system for expression of PhHV-1 gB. Recombinant (rec) baculovirus-expressed gB was identified as a 105-kDa glycosylated molecule. Proteolytic cleavage into fragments of 62 and 52 kDa was markedly delayed compared to wild-type (wt) gB. Wt and rec gB harbored endoglycosidase H (precursor)- as well as N-glycosidase F-sensitive N-glycans (proteolytic fragments). Baculovirus-expressed gB appeared to be antigenically authentic, since it was recognized in radioimmunoprecipitation and immune peroxidase monolayer assays by PhHV-1-neutralizing seal sera and by gB-specific neutralizing murine monoclonal antibodies. Furthermore, PhHV-1-neutralizing antibodies were induced in mice following immunization with baculovirus-expressed gB, indicating its suitability for incorporation in a candidate vaccine for seals. http://repub.eur.nl/res/pub/3599/ 1997-01-01T00:00:00Z The prevalence and clinical signs of phocid herpesvirus type-1 (PhHV-1) infections among harbour seals (Phoca vitulina) in a seal rehabilitation centre in the Netherlands were monitored between June and September 1993 and 1994 when 34 and 36 seals, respectively, were rehabilitated. In both years PhHV-1-related disease outbreaks occurred in the pupping season. PhHV-1 infections were diagnosed by the demonstration of a more than four-fold increase in virus neutralising serum antibodies in paired serum samples, by the isolation of the virus from swab samples in primary seal kidney cells, and by the detection of PhHV-1 DNA with a polymerase chain reaction (PCR) assay in swab samples. This assay targets a 290 bp fragment of the glycoprotein D (gD) gene equivalent of PhHV-1. The PCR assay when combined with Southern blotting (PCR-SB) was approximately 1000 times more sensitive than virus isolation when tested with serially diluted samples from PhHV-1-infected cell cultures. In contrast with virus isolation, the PCR-SB scored as positive all the animals with serological evidence of PhHV-1 infection. The majority of seals present in the centre during the outbreaks contracted the infection and developed benign upper respiratory disease. However, the severity of PhHV-1-related disease was inversely correlated with age and fatal generalised infections occurred only in neonates. http://repub.eur.nl/res/pub/3610/ 1997-01-01T00:00:00Z Blood samples were collected from 1,042 marine mammals off the coast of Alaska (USA) and Russia during the period 1978 to 1994. Eight species of pinnipeds were represented. Sera were tested for presence of neutralizing antibodies to both the PB84 isolate of phocid herpesvirus-1 (PhHV-1) and the 7848/Han90 strain of phocid herpesvirus-2 (PhHV-2). Species-specific antibody prevalences ranged from 22% to 77% for PhHV-1 and 11% to 50% for PhHV-2. Species-specific antibody prevalences for PhHV-1 were greater than or equal to prevalences for PhHV-2. For both viruses and each host species, differences in antibody prevalences were not related to: (1) sex, (2) location of capture, or (3) year of collection. Antibody prevalence of PhHV-1 in walruses (Odobenus rosmarus) could be quantitatively predicted as a function of age. These two viruses have distinct biological properties and based on current data the epizootiology of the two viruses is different, as well. No evidence of herpesvirus-induced mortality has been detected in areas included in this survey. Based on results of this survey, neither PhHV-1 nor PhHV-2 are considered significant mortality factors in mammals which inhabit the marine environment off the coast of Alaska or Russia. http://repub.eur.nl/res/pub/3563/ 1996-01-01T00:00:00Z To study the relationships between herpesvirus recently isolated from different pinniped species, antigenic and genetic analyses were performed. First, herpesviruses isolated from North American harbour seals (Phoca vitulina), a Californian sea lion (Zalophus californianus) and a European grey seal (Halichoerus grypus) were examined in an enzyme immunoassay (EIA) with a panel of monoclonal antibodies which had previously been shown to allow typing of herpesviruses from European harbour seals into two distinct virus types: phocid herpesvirus type-1 and type-2 (PhHV-1 and PhHV-2). The EIA data showed that all but one of the isolates from seals ranging in United States coastal waters were PhHV-2-like while the European grey seal herpesvirus was PhHV-1-like. Genetic characterization was facilitated by PCR analysis using primers based on conserved regions of the glycoprotein B and D (gB and gD) genes of the antigenically closely related canid (CHV) and felid (FHV) herpesvirus. Specific amplified products were obtained with five isolates antigenically characterized as PhHV-1-like but not with five PhHV-2-like isolates. Sequence analysis of the PCR products confirmed greatest similarity to members of the genus Varicellovirus of the Alphaherpesvirinae and in particular to CHV. Sequence analysis of two EcoRI fragments of the PhHV-2 genome (European isolate 7848) revealed greatest similarity to gammaherpesviruses and in particular equine herpesvirus-2. Although an unambiguous subgrouping was not feasible, this is the first evidence that PhHV-2 may be a putative gammaherpesvirus of pinnipeds. http://repub.eur.nl/res/pub/3565/ 1996-01-01T00:00:00Z Specific pathogen free (SPF) domestic cats were inoculated with tissue homogenate obtained from a Chinese leopard (Panthera pardus japonensis) that had died in a North American zoo from a natural infection with canine distemper virus (CDV). The cats developed a transient cell-associated CDV viraemia along with pronounced lymphopenia but did not show any clinical symptoms. Plasma neutralizing-antibody titres against the homologous CDV (A92-27/4, isolated from the Chinese leopard) were consistently higher than against the CDV vaccine strain 'Bussell'. The Chinese leopard CDV isolate showed in vitro biological properties reminiscent of virulent, wild-type CDV strains. Sequence analysis of the H gene of two large felid CDV isolates from the USA (A92-27/4 and A92-6) revealed up to 10% amino acid changes including up to four additional potential N-linked glycosylation sites in the extra-cytoplasmic domain as compared to CDV vaccine strains. Phylogenetic analysis was performed using the entire coding region of the H gene and a 388 bp fragment of the P gene of several morbillivirus species. Evidence was obtained that recent CDV isolates from different species in the United States (including isolates from large felids), Europe and Africa are significantly distinct from CDV vaccine strains. All wild-type CDV isolates analysed clustered according to geographical distribution rather than to host species origin. By sequence analysis a CDV epizootic among large felids in a Californian safari park was linked to a virus which most likely originated from feral non-felid carnivores. http://repub.eur.nl/res/pub/3579/ 1996-01-01T00:00:00Z The virus epizootic which resulted in significant mortality in Siberian seals (Phoca sibirica) in Lake Baikal during 1987/88 was caused by canine distemper virus. Sequence analysis of the virus glycoprotein genes revealed that it was most closely related to recent European field isolates of canine distemper virus. This paper presents evidence that the same virus continued to circulate in seals in Lake Baikal after the initial epizootic. Three out of 45 brain tissue samples collected from seals culled in the spring of 1992 were positive for canine distemper virus-specific nucleic acid by the reverse transcription/polymerase chain reaction and the sequences were closely related to that of the original virus isolated in 1988. http://repub.eur.nl/res/pub/3529/ 1995-06-13T00:00:00Z Recently an epizootic, reported to be due to a morbillivirus infection, affected the lion population of the Tanzanian Serengeti National Park. A morbillivirus phosphoprotein (P) gene fragment was amplified by PCR from tissue samples of several affected lions. Sequencing of the amplificates and subsequent phylogenetic analyses revealed that a wild-type strain of canine distemper morbillivirus (CDV) was involved. Vaccination of the local domestic dog population with proven safe CDV vaccines is proposed. http://repub.eur.nl/res/pub/3531/ 1995-01-01T00:00:00Z Sequence analysis of the haemagglutinin protein (H) gene of the morbillivirus (PDV-2) isolated from a Siberian seal (Phoca sibirica) during the 1987/1988 epizootic in Lake Baikal revealed that it was most closely related to two recent isolates of canine distemper virus (CDV) from Germany and different from CDV vaccines currently in use in that region. The virus continued to circulate in seals in Lake Baikal after the 1987/1988 epizootic since sera collected from culled seals in the spring of 1992 were positive in morbillivirus ELISA tests, reacting most strongly with the CDV antigen. http://repub.eur.nl/res/pub/3551/ 1995-01-01T00:00:00Z In recent years, serious disease outbreaks among seals and dolphins were attributed to infection with established or newly recognized morbilliviruses. The first identification of a morbillivirus as causative agent of mass mortality among marine mammals was in 1988, when the previously unrecognized phocine distemper virus (PDV) caused the death of 20,000 harbor seals (Phoca vitulina) in northwestern Europe. A similar epizootic among Baikal seals (Phoca sibirica) in Siberia in 1987 was later attributed to infection with canine distemper virus (CDV). A morbillivirus isolated from stranded harbor porpoises (Phocoena phocoena) between 1988 and 1990 proved to be yet another new member of the genus Morbillivirus, distinct from PDV and CDV and more closely related to rinderpest virus and peste-des-petits-ruminants virus: porpoise morbillivirus. A similar virus, dolphin morbillivirus, was the primary cause of mass mortality among striped dolphins (Stenella coeruleoalba) in the Mediterranean from 1990 to 1992. In this review, current knowledge of the genetic and antigenic relationships of these viruses is presented, and the origin and epizootiological aspects of the newly discovered morbilliviruses are discussed. In addition, the possible contributory role of environmental contaminant-related immunosuppression in the severity and extent of the different disease outbreaks is discussed. http://repub.eur.nl/res/pub/3501/ 1994-01-01T00:00:00Z Murine monoclonal antibodies (MAbs) were generated against phocid herpesviruses (PhHV 2557/Han88 and 7848/Han90) isolated from European harbour seals (Phoca vitulina), and against strains of both felid (FHV strain FVR 605) and canid herpesviruses (CHV isolate 5105/Han89). MAbs were characterized with respect to certain biological properties and used to outline antigenicity profiles of isolates of PhHV (n = 8), FHV (n = 7) and CHV (n = 3) in enzyme immunoassays employing fixed infected cells. A close antigenic relationship between herpesviruses derived from pinnipeds and terrestrial carnivores became evident: The majority of the MAbs was directed against epitopes which were expressed by at least two of the viral species tested. A number of MAbs detected epitopes which were conserved between all isolates of PhHV, FHV and CHV. A few MAbs recognized type-specific B-cell epitopes and facilitated the identification of single viral species. Moreover, the PhHV isolate 7848/Han90 was antigenically distinguishable both from seven other phocid herpesvirus isolates and from FHV or CHV. PhHV 7848/Han90 proved to be antigenically distinct from all other viruses tested when examined by cross neutralization utilizing various reconvalescent and hyperimmune sera. Although more data are needed to ensure that PhHV 7848/Han90 indeed is a new genuine seal herpesvirus, the preliminary clustering of two groups of phocid herpesvirus isolates, tentatively designated PhHV-1 (type isolate 2557/Han88) and PhHV-2 (represented by 7848/Han90), seems to be justified. By using selected MAbs an unambiguous identification and typing of herpesvirus isolates derived from marine mammals and terrestrial carnivores is significantly facilitated. http://repub.eur.nl/res/pub/3453/ 1992-12-01T00:00:00Z Since 1988 morbilliviruses have been increasingly recognized and held responsible for mass mortality amongst harbour seals (Phoca vitulina) and other seal species. Virus isolations and characterization proved that morbilliviruses from seals in Northwest Europe were genetically distinct from other known members of this group including canine distemper virus (CDV), rinderpest virus, peste des petits ruminants virus and measles virus. An epidemic in Baikal seals in 1987 was apparently caused by a morbillivirus closely related to CDV so that two morbilliviruses have now been identified in two geographically distant seal populations, with only the group of isolates from Northwest Europe forming a new member of the genus morbillivirus: phocid distemper virus (PDV). Because of distemper-like disease, the Baikal seal morbillivirus was tentatively named PDV-2 in spite of its possible identity with CDV. The appearance of morbilliviruses in the Mediterranean Sea causing high mortality amongst dolphins should further increase the research activities on protection strategies for endangered species of marine mammals.