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This study analyses whether the pig paramyxovirus of blue eye disease (PPBED) infects the central nervous system (CNS) utilizing anterograde and retrograde peripheral nerve transport systems. The virus was injected into muscle and skin, and inoculated per nasum. The presence of PPBED was detected by an immunohistochemical method using polyclonal mouse antibodies against the whole inactivated virus, and was revealed with polyclonal rabbit antibodies against mouse immunoglobulin G (IgG) labelled with peroxidase. The PPBED injected into the pig medial gastrocnemius (MG) muscle was detected in a terminal branch innervating the MG muscle, in neural fibres of the sciatic nerve, in fibres of the ventral and dorsal spinal roots and in ventral horn neurones of the spinal cord. When PPBED was injected into the skin area innervated by the sural nerve, it was detected in neural fibres of the sural and sciatic nerves and in spinal cord dorsal horn neurones. The per nasum inoculum rapidly invaded the CNS through the olfactory nerve. The study concluded that, in order to invade the CNS, PPBED was transported retrogradely by peripheral cutaneous and muscular nerves, and anterogradely by the olfactory nerve. No PPBED was detected in either cat peripheral nerves or in cat CNS.

Several viruses were isolated from snakes which died during an outbreak of disease in a zoo. The viruses could be cultivated at 28 degrees C on a snake cell line (VH2, ATCC CCL 140). In cell culture they formed syncytia, leading to destruction of the monolayer. The isolates were sensitive to treatment with chloroform but not to growth in the presence of IUDR and revealed haemagglutinating activity with chicken erythrocytes. These results, combined with electron microscopic examination of the infectious supernatant, confirmed the diagnosis of paramyxoviruses (PMV). Haemagglutination inhibition tests with antisera against avian PMV revealed a serological relationship between the snake viruses and the avian PMV serotypes 1 and 7. Tests for thermostability at 56 degrees C showed a marked decrease of the haemagglutination and infectivity titres in contrast to the avian PMV serotype 7. The multiplication of the snake viruses seemed to be restricted to incubation temperatures below 37 degrees C. At 28 degrees C the snake viruses could be cultivated in several cell cultures. Separation of virus proteins using polyacrylamide gel electrophoresis showed differences between the snake isolates and avian PMVs of serotypes 1 and 7.

Sera from 265 finishing pigs belonging to 70 herds, in which severe respiratory disorders were observed, were examined for antibody prevalence to porcine reproductive and respiratory syndrome virus (PRRSV), influenza virus subtypes H3N2 and H1N1, porcine respiratory corona virus (PRCV) and a recently described porcine paramyxovirus (PPMV). By immunoperoxidase-monolayer assay 69.1% of these sera were positive for PRRSV. Hemagglutination inhibiting activity was found in 55.1% of the sera for influenza virus subtype H1N1 (strain A/swine/Arnsberg/1/81). in 51.3% for influenza virus subtype H3N2 (strain A/Hong Kong/1/68) and in 14.3% for PPMV. PRCV specific antibodies, as determined by differential competitive blocking enzyme-linked immunosorbent assay were demonstrated in 192 of 236 (81.3%) sera. In order to reveal associations interspecific coefficients were calculated for antibody prevalence between PRRSV and the other viruses. Positive associations to PRRSV titres were found to PRCV, PPMV and influenza virus subtype H1N1 titres. chi-square analysis showed the statistical significance of associations regarding PRCV and influenza virus subtype H1N1. Depletion of lung macrophages after PRRSV infection is discussed as a possible mechanism for the promotion of secondary infections.

The morphology and morphogenesis of five viruses isolated in Great Britain, France and South Africa from turkeys with rhinotracheitis were examined. The five isolates were antigenically related by immunofluorescence and were indistinguishable by negative contrast and thin section electron microscopy. Negative contrast electron microscopy of infected Vero cell cultures revealed ortho- or paramyxovirus-like particles and helical nucleocapsids 14 nm in diameter with a pitch of 6 nm. The viral nature of these structures was confirmed by immuno-gold labelling, using a hyperimmune rabbit antiserum to TRT virus and 15 nm gold-labelled goat anti-rabbit IgG. Ultrastructural changes characteristics of paramyxovirus infection were observed in Vero cell cultures infected with each of the five TRT virus isolates. These alterations, which included areas of localised thickening of plasma membrane, associated cytoplasmic inclusions of nucleocapsids and budding virus particles also labelled specifically following immunogold staining. These observations are in accord with the suggestion that TRT virus is an avian pneumovirus.

Eight harbour seals (Phoca vitulina), two of them seronegative, six seropositive against PDV and a seronegative grey seal (Halichoerus grypus) were exposed to a low doses of a cell culture-propagated phocine distemper virus isolate (PDV 2558/Han 88). An intranasal route of inoculation was chosen. Clinical signs, resembling those of 1988's seal disease and seroconversion were observed in both seronegative harbour seals. One of them succumbed to the infection. The virus was not transmitted to another susceptible harbour seal which served as in-contact animal. Virus could be recovered from leucocytes of the diseased seals. Viremia was also present in a seropositive harbour seal that developed mild clinical signs; other seropositive seals were protected from clinical disease. The grey seal showed seroconversion upon inoculation, but did not develop any signs of disease. The humoral immune response of the seals plainly discriminated between homologous (PDV) and heterologous (canine distemper virus, CDV) virus as shown by virus neutralization tests and an antibody-binding assay (PLA).

Antiserum against canine distemper virus (CDV) was raised in pigs by intranasal inoculation with CDV strains CND65 and ROCKBORN. Immunoglobulin fractions were conjugated with horseradish peroxidase. Peroxidase-conjugated anti-CDV immunoglobulin preparations were used for the detection and titration of CDV, seal-derived (phocine) distemper virus (PDV) and rinderpest virus (RPV) in Vero cell cultures. For the detection and titration of corresponding neutralizing antibodies a direct neutralizing peroxidase-linked antibody (NPLA) assay was established. The results were compared with those obtained with the conventional microtitre neutralization test (MNT) based on CPE reading. In addition the sensitivity of an indirect peroxidase-linked antibody (PLA) assay was tested in parallel with that of the NPLA assay using sera obtained from CDV-immunized pigs.

As a result of a surveillance programme in North-Germany, paramyxovirus-isolates of serogroup 1 with different pathogenicity were isolated from different species of feral birds (Black-headed gull, mallard, tawny owl, tree sparrow, mute swan). In chick embryo fibroblast monolayers these isolates produced plaques exhibiting different plaque morphology. After cloning the isolates, further physicochemical, serological and pathogenicity testings were performed. On the basis of mean death time and intracerebral pathogenicity index, two of the isolates were found to be avirulent (lentogenic), whereas the others proved to be virulent (velogenic). SDS-Polyacrylamide-gel-electrophoresis showed polypeptide patterns similar to that of the PMV-1 reference strains. Summarizing all relevant findings of these investigations under consideration of the suggestions for the virological description of avian PMV (Alexander, 1987; Alexander et al., 1987; Who-Expert-Committee, 1980), including the recently established serotyping by means of monoclonal antibodies, the following designations of the PMV isolates tested herein are proposed: 1. PMV-1/black-headed gull/Germany/SSP-233/83, lentogenic NDV, monoclonal antibody group G 2. PMV-1/mallard/Germany/SSP-284/83, lentogenic NDV, monoclonal antibody group H 3. PMV-1/tawny owl/Germany/SSP-262/84, velogenic NDV, monoclonal antibody group B 4. PMV-1/tree sparrow nestling/Germany/SSP-352/84, velogenic NDV, monoclonal antibody group B 5. PMV-1/mute swan/Germany/SSP-204/85, velogenic NDV, monoclonal antibody group B.

From 16 (14%) out of 112 dead or euthanized seals originating from wildlife and seal orphanages phocine morbillivirus was isolated. The majority of viral isolates in cell cultures was obtained from lung homogenates of 15 out of 71 free-ranging seals (21%). The virus was isolated by longterm cultivation in roller cultures of seal kidney cells. The phocine morbillivirus was detected by typical cytopathogenic alteration and by peroxidase-linked antibody (PLA) assay, respectively. A neutralization test based on PLA was used for antibody detection in seals using a canine distemper virus (CDV) strain and in parallel one of the phocine morbillivirus isolates. All sera tested were proven to contain neutralizing antibodies of higher titres against the latter virus than against the CDV strain. Several seals furnished morbillivirus isolates and at the same time exhibited neutralizing antibodies of low to medium titres. No viral isolates were obtained from the majority of sick animals with moderate to high neutralizing titres (greater than 1/1,000). The significance of these findings is discussed in relation to the cause of the mass mortality amongst seals observed in 1988 in the Bay of Heligoland.

Of 57 seals hospitalized in the Norddeich Orphanage, 37 (65%) had died until 22nd of August, 1988, when the first collection of blood samples from the survivors commenced. All the sera including those obtained at a later date from the remaining animals had invariably higher neutralizing antibody titres against a phocine distemper virus (PDV) isolate than against canine distemper virus (CDV). The difference of mean titres was calculated to be more than 1.5 x log10 serum dilution. Peak titres demonstrated by a direct neutralization peroxidase-linked antibody (NPLA) assay reached 1/90,000. Attempts to isolate PDV in seal kidney cell cultures from heparinized blood samples collected from convalescent animals were not successful. From the increase in antibody titre following the last fatal case it was concluded that the devastating epidemic sweeping through the Norddeich Orphanage was primarily due to phocine distemper.

The recent review is based on recent literature on the epizootiology and on properties of avian paramyxoviruses in vivo and in vitro. Paramyxoviruses are classified according to their pathogenicity (especially serotype 1), in addition to serotyping by means of hemagglutination-inhibition tests and by monoclonal antibody binding assays. Also, neuraminidase-inhibition tests, double-immunodiffusion tests can be performed. Growth properties of the different paramyxovirus-serotypes are accessory techniques to classify paramyxoviruses parallel to the determination of the pathogenicity. Structural polypeptides can be demonstrated in polyacrylamide-electrophoresis, thus leading to a distinction of the different serotypes.