The serological response of the common seal (Phoca vitulina) and the grey seal (Halichoerus grypus) to phocine distemper virus as measured by a canine distemper virus neutralisation test.

The application of a canine distemper virus (CDV) neutralisation test revealed that antibody titres of up to 11,620 were present in the sera of common seals during the epizootic of phocine distemper in British waters. However, during the later stages of the epizootic and afterwards, a surprisingly high proportion of animals were still seronegative. Titres of from 90 to 183 were common in diseased animals and were probably indicative of partial immunosuppression. A few animals with proven phocine distemper had titres of less than 64, this being evidence of a more profound immunosuppression. Recovered animals tended to have moderate to high titres and in some cases these persisted for at least 6.5 months. By contrast, the administration of an inactivated CDV vaccine produced antibody which declined rapidly to very low levels within the same period. Infection also occurred in grey seals in 1988 but without severe mortality. Antibody titres in this species nearly always lay in the range 128-734 and persisted at or around this level for at least 12 months. In both common and grey seals, antibody was transferred by suckling from mother to pup but absorption by the pup seemed to take place more slowly than in domestic mammals.

Serologic study of phocine distemper in a population of harbor seals in Scotland.

A serologic survey of the prevalence of morbillivirus antibodies was conducted in a population of harbor seals (Phoca vitulina) from northeastern Scotland, where mortality was comparatively low during the 1988 phocine distemper virus outbreak. None of the 12 seals sampled before the epizootic were seropositive. Thirty-five (52%) of 68 seals sampled after the beginning of the epizootic were seropositive, although there were significant age-related differences in both the number of seropositive individuals and in antibody levels. Marking studies showed that most seropositive seals caught during the peak of the epizootic survived for several months. Thus, the low mortality observed in this population did not appear to result from a lack of contact with the virus.

Distemper encephalitis in pups after vaccination of the dam.

A five-year-old labrador bitch which had whelped 10 pups three days previously was given booster vaccination against distemper, adenovirus, parvovirus, parainfluenzavirus and leptospirosis. Eighteen days later, signs of central nervous system disease developed in some of the pups, five of which were ultimately euthanased. The cause of the nervous disease was found to be canine distemper, and serological studies showed that the infection was limited to some members of the litter, suggesting that the vaccinal rather than a field virus was more likely to have been responsible.

Studies on canine parvovirus infection: preparation of challenge virus.

Two techniques, adsorption on to hydroxylapatite and density gradient centrifugation, were investigated as prospective methods for the large scale purification of canine parvovirus from faecal suspensions. Adsorption with hydroxylapatite successfully removed virus from faecal material. However, the resultant virus was contaminated and some virus was left behind in the faecal suspension. Repeated adsorption with hydroxylapatite appeared to result in some damage to the virus particles. In contrast, density gradient centrifugation provided a simple, economical method of purification which yielded uncontaminated, infectious virus. The final method, using both isopyknic and rate zonal centrifugation is described.

Canine parvovirus: interaction between passive immunity and virulent challenge.

Two groups of puppies, one passively immunised by the administration of hyperimmune serum and the other with natural maternally derived antibody, were inoculated orally with virulent canine parvovirus of faecal origin. Serum antibody titres declined more rapidly in both groups after challenge than before. The dogs became clinically affected but the onset of clinical signs, seroconversion and faecal excretion of virus was delayed when compared to controls. It is postulated that this rapid decline of antibody was due to its sequestration by virus after the initial phase of viral replication in the lymphoid tissues. These findings have important implications. The incubation period of the disease is prolonged, making it more difficult to estimate accurately the time of infection in clinically affected animals. Furthermore, the more rapid decline of maternally derived antibody, which could occur in endemically infected premises, may complicate immunisation programmes based on the isolation and segregation of puppies in anticipation of a predicted decline in maternally derived antibody before vaccination.

Studies of parvovirus vaccination in the dog: the performance of live attenuated feline parvovirus vaccines.

The performance of three live attenuated feline parvovirus vaccines licensed for use in the dog was studied. At the end of the primary vaccination course 67 per cent of dogs had inadequate antibody levels (less than or equal to 32) as measured by a haemagglutination inhibition test. Interference by maternal antibody accounted for some of the failures but the fact that there was no significant difference in performance between dogs vaccinated at 12 weeks or 16 weeks of age indicated that maternal antibody was not the only factor.

Encephalitis in dogs associated with a batch of canine distemper (Rockborn) vaccine.

During a period of seven months in 1982-83 cases of postvaccinal encephalitis were recorded in dogs in various parts of Britain after the administration of a particular batch of combined distemper/hepatitis vaccine. Detailed investigations of one of these cases revealed that the distemper component was responsible and the vaccine virus was recovered from the brain of an affected dog.

Canine parvovirus enteritis 2: Pathogenesis.

Two groups of puppies, eight and 10 weeks of age, were inoculated orally with canine parvovirus of faecal origin. The patterns of faecal excretion of virus, antibody production and systemic viral localisation following inoculation were studied. Faecal excretion of virus was first apparent at day 3 after inoculation, was present most frequently and in greatest quantity at days 4 to 7 after inoculation and fell sharply thereafter. Serum antibody was first detected at day 5 after inoculation with high titres in all samples from day 7 onwards. Virus isolation from serum samples revealed a non-cell associated viraemia at days 3 and 4 after inoculation. Immunocytochemical examination, using both immunofluorescence and immunoperoxidase techniques, first revealed antigen in the thymic cortex at day 1 after inoculation and in the germinal centres of the lymph nodes and the splenic white pulp from days 2 and 3. Viral antigen was first detected in the intestines at day 4 in individual cells in the proliferative zone of the crypt epithelium. From day 5 onwards, the amount of antigen present in the lymphoid tissue decreased so that by days 7 and 8, only a trace was present. There was widespread specific staining in the small intestinal mucosa at day 6, but little antigen was present by day 7. Virus was present in the bone marrow of some dogs killed at days 5 and 6.

Canine parvovirus enteritis 3: Scanning electron microscopical features of experimental infection.

A group of 10-week-old puppies was orally inoculated with canine parvovirus of faecal origin. Scanning electron microscopy was used to study and compare the surface topography in both control and inoculated animals. In control dogs the villi were tall and finger-like in shape and numerous irregular transverse circumferential grooves were present on the surface. At higher magnification, the outlines of individual epithelial cells and depressions, interpreted as goblet cells, could be discerned. In the inoculated dogs, scanning electron microscopy changes were first seen at six days after inoculation. The small intestinal mucosa was covered by a thick layer of mucus. The underlying villi were stunted and had lost their surface features. In some instances there was loss of the luminal epithelium, exposing the lamina propria. In addition, there was dilation of the circumvillar basins and the crypt mouths. There appeared to be regenerative changes by day 7 after inoculation. The surface of the small intestinal mucosa was still covered by a thick layer of mucus. Where villi could be discerned, they were short and pointed and transverse grooves could be seen on their surface. There was some hypertrophy of the intervillus ridges. The changes in the surface topography of the small intestinal mucosa following canine parvovirus infection are compared to those seen in enteric infections in other species and the similarity of the lesion to that seen following sublethal irradiation is discussed.

Canine parvovirus enteritis 1: Clinical, haematological and pathological features of experimental infection.

The effect of oral infection of puppies, eight and 10 weeks old, with canine parvovirus of faecal origin was studied. Clinical signs of enteric disease were first apparent at five days after inoculation and persisted during days 6 and 7 after inoculation. The severity of clinical signs varied from transient dullness and anorexia to emesis, dysentery and death. Changes in haematological parameters were first found at day 3 after inoculation when a relative lymphopenia was observed. A profound neutropenia developed in severely affected dogs after the appearance of clinical enteric disease. Post mortem examination revealed thymic atrophy in all dogs killed on day 4 after inoculation. Macroscopic changes in the small intestine were apparent only in animals examined during the phase of severe enteric disease and consisted of thickening, rigidity and congestion of the small intestines. Microscopically there was lymphocytolysis in the thymic cortex and the germinal centres of the lymph nodes from days 2 and 3 after inoculation respectively and this rapidly resulted in depletion of these tissues. There was repopulation of lymph nodes from day 7 after inoculation but significant thymic regeneration was not apparent during the course of this study. In the small intestine, necrosis of crypt epithelium, atrophy of villi and, in some areas, complete collapse of mucosal architecture were found but the extent of these changes varied along the length of the small intestine and between individuals. Regenerative intestinal changes were observed in those animals surviving the acute phase of enteric dysfunction. The variable severity of clinical and enteric lesions, together with the factors which may affect the expression of clinical disease, are discussed.

Immunity to canine adenovirus respiratory disease: effect of vaccination with an inactivated vaccine.

Nine puppies without maternal antibody to canine adenovirus (CAV) were divided into two groups. The first consisted of six puppies, each of which was given two doses of a commercial inactivated CAV-1 vaccine, 14 days apart. Eight days after administration of the second dose of vaccine, all six puppies, together with the second group, consisting of three unvaccinated controls, were challenged with an aerosol of virulent CAV-2. One dog from each group was killed on the third, fifth and 10th days after challenge and the three additional vaccinates killed at intervening times. All of the dogs developed respiratory signs, mainly coughing and tachypnoea, but the vaccinated dogs made a more rapid recovery. The lungs of both groups were consolidated, the areas affected being more extensive in the controls, and histological examination revealed the main lesion to be a severe necrotising bronchiolitis. Virus was isolated from the respiratory tissues and from throat swabs collected from both groups of dogs. The presence of neutralising antibody in the serum was not, of itself, sufficient to control viral replication and oblate the disease.

Experimental canine adenovirus glomerulonephritis: histological, immunofluorescence and ultrastructural features of the early glomerular changes.

Eighteen 16-week-old dogs were inoculated i.v. with a virulent strain of canine adenovirus and the renal glomeruli were examined by light, electron (transmission and scanning) and immunofluorescence microscopy at intervals from 24 h to 7 days after inoculation of virus. From 2 days onwards intranuclear inclusion bodies and cell-associated viral antigen were detected in mesangial, endothelial and circulating mononuclear cells. This was accompanied by swelling and, in some cases, necrosis of these cells with partial or complete occlusion of capillary loops. In 5 dogs examined on days 5-7 after inoculation of virus, granular deposition of IgG, C3 and viral antigen was observed in mesangial areas and electron dense deposits were found in the mesangial matrix.

Patterns of cilia formation in the lower respiratory tract of the dog: a scanning electron microscopic study.

A scanning electron microscopic study of the distribution of ciliated cells in the tracheobronchial tree of 25 dogs whose ages ranged from four hours to six months was carried out. In newborn puppies, only the dorsal wall of the trachea was completely ciliated while the lateral and ventral walls showed patchy ciliation. This pattern of cilia formation persisted until five days of age when the whole tracheal wall was found to be completely ciliated. The bronchus of newborn puppies was uniformly poorly ciliated but almost complete ciliation was achieved by two days. Likewise, large and small bronchioles of newborn animals had few cilia and, although the number of ciliated cells had increased by two days, complete ciliation was never observed regardless of the age of the dog. No ciliated cells were found in the respiratory bronchioles of any of the 25 dogs.

Immunity to canine adenovirus respiratory disease: a comparison of attenuated CAV-1 and CAV-2 vaccines.

Four litters of puppies were divided into three groups. One group was vaccinated with a live CAV-1 vaccine and another with a live CAV-2 vaccine. Throat swabs were collected from two dogs in each of these groups to monitor the possible excretion of vaccine virus, but none was found. Both groups, together with the third group of unvaccinated controls, were challenged 17 days later with an aerosol of virulent CAV-2. One dog from each group was killed on the third, fourth, seventh, ninth, 11th and 14th days after challenge. The unvaccinated dogs developed a clinical disease characterised by anorexia, dullness, coughing and tachypnoea. The lungs were consolidated and histological examination revealed the main lesion to be a severe necrotising bronchiolitis. Large amounts of virus were present in the respiratory tissues of these dogs and high titres of virus were isolated from throat swabs. In contrast, both groups of vaccinated dogs remained clinically almost normal with minimal lesions, present for a much shorter period of time. Virus was found on day 4 in the respiratory tissues of one dog vaccinated with CAV-1 but the other vaccinated animals contained little or no virus. In general, the degree of protection afforded by CAV-1 vaccine seemed similar to that provided by CAV-2 vaccine.