Canine parvovirus type 2 vaccine protects against virulent challenge with type 2c virus

The ability of dogs vaccinated with a live attenuated CPV type 2 (Nobivac Intervet) vaccine to resist challenge with a current CPV2c isolate was investigated. Six SPF beagle dogs were given the minimum recommended course of vaccination, comprising a single inoculation of vaccine (Nobivac Lepto+Nobivac Pi) at 8-10 weeks of age followed 3 weeks later with a parvovirus vaccine in combination with distemper, adenovirus and parainfluenza virus (Nobivac DHPPi) and a repeat leptospirosis vaccine. Six control dogs were kept unvaccinated. All animals were challenged orally with a type 2c isolate of CPV and monitored for clinical signs, virus shedding, white blood cell fluctuations and serological responses. All vaccinated dogs were fully protected; showing no clinical signs nor shedding challenge virus in the faeces, in contrast to control animals, which displayed all the typical signs of infection with pathogenic CPV and shed challenge virus in the faeces.     

Protection against feline leukemia virus challenge for at least 2 years after vaccination with an inactivated feline leukemia virus vaccine

Twelve cats were vaccinated at 8 and 11 weeks of age with a commercially available inactivated FeLV vaccine (Nobivac FeLV, Intervet/Schering-Plough Animal Health). Eleven cats served as age-matched, placebo-vaccinated controls. All cats were kept in isolation for 2 years after vaccination and were then challenged with virulent FeLV to evaluate vaccine efficacy and duration of immunity. Cats were monitored for 12 weeks after challenge for development of persistent viremia using a commercial FeLV p27 ELISA. Persistent viremia developed in all 11 (100%) of the control cats, whereas 10 of 12 (83%) vaccinated cats were fully protected from persistent viremia following challenge. The results demonstrate that the vaccine used in this study protects cats from persistent FeLV viremia for at least 2 years after vaccination.     

Prevention of renal infection and urinary shedding in dogs by a Leptospira vaccination

Prevention of urinary shedding of Leptospira interrogans spp. by chronically infected dogs remains a key objective of the vaccination in dogs against leptospirosis which is a zoonotic disease. An inactivated bivalent vaccine composed of Leptospira interrogans serovars icterohaemorrhagiae [L. icterohaemorrhagiae] and canicola [L. canicola] bacterins was tested for its ability to protect puppies against a challenge exposure with L. icterohaemorrhagiae. The vaccine was administered twice at a 3-week interval to six puppies aged from 8 to 9 weeks. Six other puppies were used as unvaccinated controls. All puppies were challenged 2 weeks after the second vaccine injection by intraperitoneal (IP) administration of L. icterohaemorrhagiae (day 0). Clinical signs, haematological and biochemical changes and evidence of Leptospira in blood, urine and kidney were monitored for 4 weeks after the challenge exposure (days 0-28). Puppies were euthanised on day 28 for post-mortem and histological examinations of liver and kidney. Control group presented clinical pictures of severe or subclinical infection. One dog developed severe clinical signs (hypothermia, depression, anorexia, abdominal pain, dehydration, icterus, weight loss) and died on post-infection day (PID) 7 due to an acute renal failure. Gross and microscopic lesions were in accordance with this clinical pattern. In the five remaining control dogs, the challenge exposure induced mainly a systemic infection including leptospiraemia, leptospiruria and renal carriage. The vaccinated group remained healthy throughout the study period. In conclusion, immunisation with a Leptospira vaccine was shown to protect dogs against symptomatology and leptospiraemia, urine shedding and renal infection.     

Comparison of antibody responses after vaccination with two inactivated rabies vaccines

Thirty laboratory dogs were randomly assigned to two groups (A and B) of 15 dogs and subcutaneously vaccinated with a single dose of one of two commercially available monovalent inactivated rabies vaccines: RABISIN (Merial, France) (group A) and NOBIVAC Rabies (Intervet International) (group B). Rabies antibodies were measured over a period of 4 months using the fluorescent antibody virus neutralization (FAVN) test. The two vaccines performed differently in terms of magnitude and persistence of rabies antibodies titers in dogs. Two weeks after vaccination, average rabies antibody titers peaked at 2.53 IU/mL (range, 0.17-13.77 IU/mL) and 1.26 IU/mL (range, 0.50-4.56 IU/mL) in groups A and B dogs, respectively. The average FAVN antibody titres against rabies on D28, D56, D84, D112 and D120 were significantly higher in group A than in group B. Although all dogs from group B serologically responded to vaccination, the proportion of dogs with antibody titres >or=0.5 IU/mL dropped significantly after D28 and was statistically significantly lower on D56, D84 and D112 compared to group A dogs. In conclusion, in the context of international trade, the choice of the vaccine and the timing of blood tests are critical factors in achieving successful serological test results after rabies vaccination. RABISIN induces high and sustained antibody titres against rabies, increasing the flexibility for the time of blood sampling after primo-vaccination.     

Canine parainfluenza-Bordetella bronchiseptica vaccine immunogenicity

The immunogenicity and safety of 3 serials of a canine parainfluenza (CPI) virus-Bordetella bronchiseptica vaccine was evaluated. Each serial was used to vaccinate 10 dogs with single doses given intranasally. The 30 vaccinated and 10 nonvaccinated controls dogs were challenge exposed with aerosols of virulent CPI virus and B bronchiseptica at 18 days and at 21 days, respectively, after vaccination. After challenge exposure, none of the 30 vaccinated dogs had clinical signs of disease; however, 9 of the 10 nonvaccinated dogs developed coughing problems. The CPI virus was isolated from nasal swab specimens obtained from nonvaccinated dogs on an average of 5.1 days after challenge exposure, but was not isolated from any of the specimens obtained from the vaccinated dogs. Bordetella bronchiseptica was isolated from nasal swab specimens obtained from both vaccinated and nonvaccinated dogs up to 18 days after challenge exposure. The erythrocyte sedimentation rates and total leukocyte counts for control dogs were generally increased, in contrast to those for the vaccinated groups. Dogs showed a primary serologic response to CPI virus and B bronchiseptica after vaccination and an anamnestic response to the bacterium after challenge exposure. Adverse local or systemic reactions attributable to the bivalent vaccine were not observed in the vaccinated dogs.     

Protection of vaccinated pigs against experimental infections with homologous and heterologous Haemophilus parasuis

The efficacy of a new Haemophilus parasuis vaccine for pigs was investigated. The vaccine contains H parasuis serotype 5 cells and is adjuvanted with Diluvac Forte (Intervet). Groups of pigs were vaccinated at five and seven weeks with 2 ml and their littermates served as unvaccinated controls. The vaccinated pigs were protected against a challenge with another strain of Hparasuis serotype 5 at two, eight and 17 weeks after the second vaccination, whereas the controls became very ill. The susceptibility of the pigs to the infection decreased with increasing age. After a heterologous challenge with H parasuis serotypes 1, 12, 13 and 14, two weeks after the second vaccination, the vaccine also gave clear protection. The severity of the illness among the control pigs differed with the different serotypes.     

Three-year duration of immunity in dogs following vaccination against canine adenovirus type-1, canine parvovirus, and canine distemper virus

A challenge-of-immunity study was conducted to demonstrate immunity in dogs 3 years after their second vaccination with a new multivalent, modified-live vaccine containing canine adenovirus type 2 (CAV-2), canine parvovirus (CPV), and canine distemper virus (CDV). Twenty-three seronegative pups were vaccinated at 7 and 11 weeks of age. Eighteen seronegative pups, randomized into groups of six dogs, served as challenge controls. Dogs were kept in strict isolation for 3 years following the vaccination and then challenged sequentially with virulent canine adenovirus type 1 (CAV-1), CPV, and CDV. For each viral challenge, a separate group of six control dogs was also challenged. Clinical signs of CAV-1, CPV, and CDV infections were prevented in 100% of vaccinated dogs, demonstrating that the multivalent, modified-live test vaccine provided protection against virulent CAV-1, CPV, and CDV challenge in dogs 7 weeks of age or older for a minimum of 3 years following second vaccination.     

Comparison of the mucosal immune response in dogs vaccinated with either an intranasal avirulent live culture or a subcutaneous antigen extract vaccine of Bordetella bronchiseptica

Healthy dogs with low antibody titer to Bordetella bronchiseptica were vaccinated intranasally with an avirulent live vaccine, subcutaneously with an antigen extract vaccine, or subcutaneously and intranasally with a placebo. Intranasally vaccinated dogs developed B. bronchiseptica-specific IgA titers in nasal secretions that remained at high levels until the end of the study; dogs vaccinated subcutaneously with the antigen extract or placebo did not develop measurable antigen-specific IgA titers in nasal secretions. Dogs were challenged with virulent live B. bronchiseptica 63 days after vaccination. Intranasally vaccinated dogs had significantly lower cough scores (P < or =.0058) and shed significantly fewer challenge organisms (P <.0001) than dogs in either of the other groups. Cough scores of subcutaneously vaccinated dogs were not significantly different from placebo-vaccinated dogs.     

Canine distemper virus neutralising antibodies in vaccinated dogs

The associations between the levels of canine distemper virus neutralising antibodies and vaccination history, age and gender were investigated in a cross-sectional study of a sample of 4627 dogs from the Finnish urban dog population. Dogs vaccinated with either Canlan (Langford Laboratories) or Dohyvac (Solvay Animal Health) or with both, had significantly lower titres than those vaccinated with Candur (Behringwerke), Duramune (Fort Dodge Laboratories) or Nobivac (Intervet), or with combinations including at least one of these. The vaccines were classified as having low and high immunogenicity on the basis of the geometric mean titre achieved by the vaccine when compared with the geometric mean titre of the entire dataset. The low geometric mean titre of Canlan, Dohyvac and their combination groups resulted from the large proportion of dogs without detectable titres, especially dogs under one year of age, rather than from uniformly low titres. An age-stratified comparison of vaccine usage and titres showed that the division of the vaccines into low and high immunogenicity vaccines was apparent in the dogs less than two years of age but not in the older dogs. The first vaccination with the high immunogenicity vaccines resulted in a higher proportion of dogs with detectable antibodies than even repeated vaccination with the low immunogenicity vaccines. Neither the time elapsed since the most recent vaccination nor the gender of the dogs was associated with the titre of antibody.     

Vaccines against Aujeszky's disease: Evaluation of their efficacy under standardized laboratory conditions

Summary

A standardized test was developed to compare the efficacy of Aujeszky's disease virus (ADV) vaccines under laboratory conditions. Per test 3 groups of 6 to 8 sero‐negative pigs were used. The first vaccination was done at 10 weeks of age. One group was vaccinated once, another was vaccinated twice and the 3rd served as control. Pigs were challenge exposed to the virulent NIA‐3 strain of ADV 12 weeks after the first vaccination. Apart from mortality, average periods of growth arrest, fever and virus shedding after challenge were used as parameters to evaluate vaccine efficacy.

Two inactivated and 4 attenuated vaccines were tested. Two attenuated vaccine viruses were excreted after vaccination. Despite maximal standardization, a considerable variation still existed between the experiments in mortality and growth arrest periods of control pigs after challenge. However, the controls were always more severely affected than the vaccinated pigs. All vaccines except one were effective in preventing death after challenge, but none conferred complete protection. Most vaccinated pigs still lost weight, developed fever and shed virus after challenge. Revaccination after 3 or 4 weeks had little effect, particularly with the attenuated vaccines. The results of the present study indicate that 2 of the attenuated vaccines conferred the best protection, I attenuated vaccine appeared to be as effective as the 2 inactivated ones, and the 4th attenuated vaccine was least effective.     

Prevention of renal carriage of leptospirosis in dogs by vaccination

Dogs vaccinated with a Leptospira interrogans vaccine containing serogroups Canicola and Icterohaemorrhagiae and prepared from cultures grown in a protein-free medium resisted challenge with heterologous representatives of these serogroups. In contrast, control dogs were pyrexic and leptospires were isolated from the blood for nine days following Canicola challenge and six days after Icterohaemorrhagiae challenge. Leptospires were isolated from the urine of controls throughout the post challenge period and from kidneys at post mortem examination of six out of six and four out of six Canicola and Icterohaemorrhagiae challenged controls, respectively. There have been no reports of hypersensitivity reactions to this vaccine since its introduction in 1980.     

High-cell-passage canine coronavirus vaccine providing sterilising immunity

OBJECTIVES: To evaluate the ability of a high-cell-passage canine coronavirus vaccine to immunise dogs against challenge with a field isolate of the virus. METHODS: Three dogs that had previously tested seronegative and virus-negative for canine coronavirus were inoculated twice, at 21-day intervals, with the vaccine and kept under observation. Two seronegative and virus-negative dogs served as unvaccinated controls. For safety tests, two additional dogs were inoculated oronasally with 10 times the vaccinal dose and no reactions were observed. Faecal samples were collected daily from the vaccinated dogs after the first and second inoculations. Both vaccinated and control dogs were challenged two weeks after the second vaccination with a field canine coronavirus strain. Blood samples were collected for serological tests before vaccination and at weekly intervals after vaccinations and challenge. RESULTS: Virus was not detected in faecal samples after the first or second vaccinations by virus isolation assays and PCR. Significantly, the vaccinated dogs did not have clinical signs after challenge and no virus shedding was observed. The two unvaccinated control dogs had moderate enteritis, and virus was detected in cell cultures starting from three days postchallenge (dog 1) and two days postchallenge (dog 2), and by PCR for 23 median days. CLINICAL SIGNIFICANCE: This study showed the efficacy of a high-cell-passage canine coronavirus vaccine in preventing infection of dogs by virulent virus and, specifically, its ability to induce sterilising immunity.     

Duration of immunity in dogs vaccinated against leptospirosis with a bivalent inactivated vaccine

Duration of immunity in dogs induced with current commercial inactivated leptospirosis vaccines and evaluated against experimental infection, to date, has hardly been documented. The purpose of the present work was to assess the duration of immunity in dogs that is attainable with a commercial inactivated bivalent leptospirosis vaccine. For this purpose, young dogs were vaccinated twice followed by challenge with either Leptospira interrogans serovar canicola or L. interrogans serovar icterohaemorrhagiae 5 weeks, 27 weeks or 56 weeks after the second vaccination. For assessment of the duration of immunity, titres of agglutinating serum antibodies were measured before and after challenge, and the effects of challenge on a variety of parameters were determined including reisolation of challenge organisms from blood, urine and kidney. Both challenge strains induced a generalised infection in control dogs, the canicola strain being most virulent. From the results with different parameters it appeared that the two vaccinations induced a high rate of protection from generalised infection with canicola and icterohaemorrhagiae at 5, 27 and 56 weeks after the second vaccination. In addition, after 56 weeks, still a high level of immunity against renal infection with sv. canicola and, as a consequence, urinary shedding of sv. canicola bacteria, was demonstrated. It was, therefore, concluded that with this vaccine, using this vaccination schedule, a duration of immunity of 1 year can be attained against infection with both serovars.     

Use of a monovalent leptospiral vaccine to prevent renal colonization and urinary shedding in cattle exposed to Leptospira borgpetersenii serovar hardjo.

OBJECTIVE: To determine whether a monovalent Leptospira borgpetersenii serovar hardjo (type hardjobovis) vaccine commercially available in Australia, New Zealand, Ireland, and the United Kingdom would protect cattle from renal colonization and urinary shedding when exposed to a US strain of Leptospira borgpetersenii serovar hardjo. ANIMALS: 24 Hereford heifers that lacked detectable antibodies against serovar hardjo. PROCEDURE: Heifers received 2 doses, 4 weeks apart, of the commercial hardjo vaccine (n = 8) or a monovalent US reference hardjo vaccine (8) or were not vaccinated (controls; 8). Heifers were challenged 16 weeks later by intraperitoneal inoculation or conjunctival instillation. Serum antibody titers were measured weekly, and urine samples were examined for leptospires. Heifers were euthanatized 11 to 14 weeks after challenge, and kidney tissue was examined for evidence of colonization. RESULTS: All 8 heifers vaccinated with the reference vaccine were found to be shedding leptospires in their urine and had evidence of renal colonization. All 4 control heifers challenged by conjunctival instillation and 2 of 4 control heifers challenged by intraperitoneal inoculation shed leptospires in their urine, and all 8 had evidence of renal colonization. In contrast, leptospires were not detected in the urine or tissues of any of the 8 heifers that received the commercial hardjo vaccine. Heifers that received the commercial hardjo vaccine had significantly higher antibody titers than did heifers that received the reference vaccine. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that cattle that received 2 doses of the commercial hardjo vaccine were protected against renal colonization and urinary shedding when challenged with L borgpetersenii serovar hardjo strain 203 four months after vaccination.     

Vaccines against Aujeszky's disease: evaluation of their efficacy under standardized laboratory conditions

A standardized test was developed to compare the efficacy of Aujeszky's disease virus (ADV) vaccines under laboratory conditions. Per test 3 groups of 6 to 8 sero-negative pigs were used. The first vaccination was done at 10 weeks of age. One group was vaccinated once, another was vaccinated twice and the 3rd served as control. Pigs were challenge exposed to the virulent NIA-3 strain of ADV 12 weeks after the first vaccination. Apart from mortality, average periods of growth arrest, fever and virus shedding after challenge were used as parameters to evaluate vaccine efficacy. Two inactivated and 4 attenuated vaccines were tested. Two attenuated vaccine viruses were excreted after vaccination. Despite maximal standardization, a considerable variation still existed between the experiments in mortality and growth arrest periods of control pigs after challenge. However, the controls were always more severely affected than the vaccinated pigs. All vaccines except one were effective in preventing death after challenge, but none conferred complete protection. Most vaccinated pigs still lost weight, developed fever and shed virus after challenge. Revaccination after 3 or 4 weeks had little effect, particularly with the attenuated vaccines. The results of the present study indicate that 2 of the attenuated vaccines conferred the best protection, 1 attenuated vaccine appeared to be as effective as the 2 inactivated ones, and the 4th attenuated vaccine was least effective.     

The duration of immunity to an inactivated adjuvanted canine parvovirus vaccine. A 52 and 64 week postvaccination challenge study

Dogs were successfully isolated for a period of either 52 or 64 weeks following vaccination with an inactivated, adjuvanted canine parvovirus-2 vaccine. Antibody persisted in all ten vaccinated dogs, although in one case by 52 weeks postvaccination only virus neutralizing antibody, and not hemagglutination-inhibiting antibody, could be detected. Sentinel unvaccinated dogs housed alongside the vaccinated dogs throughout the study remained free of canine parvovirus-2 antibody until challenged. Upon oral challenge with canine parvovirus-2 infected material all unvaccinated dogs developed one or more signs of canine parvovirus-2 disease, shed virus and developed antibody. None of the vaccinated dogs became overtly sick. Of the five vaccinated dogs challenged 52 weeks after vaccination, three shed virus and one showed a significant rise in antibody. At 64 weeks after vaccination only one of the five challenged dogs shed virus and showed a boost in antibody titer.     

Immune response of sows and their offspring to pseudorabies virus: serum neutralization response to vaccination and field virus challenge.

One month prior to breeding, sows were vaccinated with an attenuated pseudorabies virus vaccine or challenged with a field strain of pseudorabies virus. A third group of sows were not vaccinated or challenged before breeding. Pigs from these sows were vaccinated at 3, 6, or 12 weeks of age and challenged with virulent virus three weeks later. One pig from each litter served as an unvaccinated, unchallenged control. Serum neutralization titers of these pigs were monitored from birth until 22 weeks of age. Titers of the sows were monitored through breeding, gestation and farrowing. The maximum prefarrowing anti-pseudorabies virus titer in the field virus challenged sows occurred four weeks following challenge. A significant decline in titers occurred at farrowing. Titers rose from one week postfarrowing and then declined. Titers in the field virus infected sows were consistently two to threefold greater than those of the vaccinated sows. The maximum prefarrowing anti-pseudorabies virus titer in the vaccinated sows occurred six weeks following vaccination. The geometric mean titer in these sow's then decreased and increased for two weeks after farrowing. The results in the pigs can be summarized as follows: Pigs from control sows had a greater serological response following field virus challenge than following vaccination with a modified live virus. Pigs from control sows responded serologically to vaccination at 3, 6 and 12 weeks of age. Pigs from control sows which were challenged at 6, 9 and 15 weeks of age had similar antibody responses. Pigs from vaccinated sows had no increase in titer following vaccination at three and six weeks of age. Titers increased when these pigs were vaccinated at 12 weeks of age. There was no significant increase in mean titers of pigs from challenged sows following vaccination at 3, 6 and 12 weeks of age. Vaccinated pigs from control and vaccinated sows had a secondary response following challenge three weeks after vaccination.(ABSTRACT TRUNCATED AT 250 WORDS)     

Comparison between immune responses and resistance induced in BALB/c mice vaccinated with RB51 and Rev. 1 vaccines and challenged with Brucella melitensis bv. 3

BALB/c mice were immunized with live rough Brucella abortus RB51 or smooth Brucella melitensis Rev. 1 vaccines and challenged with a B. melitensis field strain. Protection was assessed by a variety of serological tests and recovery of vaccinal and challenge strains by culture. Mice vaccinated with RB51 gave negative results in the conventional serological tests prior to challenge, namely; standard tube agglutination test (SAT), Rose Bengal plate test (RBPT), buffered acidified plate antigen test (BAPAT), and mercaptoethanol test (MET). Sero-conversion took place to a whole-cell bacterial buffered RB51 antigen after vaccination and persisted for 7 weeks post-vaccination. Mice challenged with B. melitensis were assessed for bacterial load and immune response for 12 weeks after challenge. Protection units were showed that Rev. 1 vaccine was superior to RB51 vaccine in protection of mice against B. melitensis. However, RB51 vaccine has the advantage that it would not elicit antibodies to standard serological tests based on the LPS O antigen. RB51 vaccine could therefore be used for control of B. melitensis infection and avoid confusion in the use of standard sero-diagnostic tests.     

Serologic responses of dogs given a commercial vaccine against Leptospira interrogans serovar pomona and Leptospira kirschneri serovar grippotyphosa

OBJECTIVE: To evaluate serum titers obtained by use of the microscopic agglutination test (ie, MAT titers) to Leptospira interrogans serovar pomona and autumnalis and Leptospira kirschneri serovar grippotyphosa in dogs given a commercial vaccine against serovars pomona and grippotyphosa. ANIMALS: Forty 12-week-old puppies and 20 mature Beagles. PROCEDURE: Puppies received a commercial vaccine against serovars pomona and grippotyphosa at 12 weeks of age, then received a booster vaccine and 3 weeks later; mature dogs received the vaccine once. Serum MAT titers to serovars pomona, autumnalis, and grippotyphosa were measured before vaccination and at 2, 4, 6, 10, and 16 weeks after the first or only vaccination. RESULTS: Of the 40 puppies vaccinated, 40, 0, and 40 developed MAT titers of > 100 after vaccination to serovars pomona, grippotyphosa, and autumnalis, respectively. Microscopic agglutination test titers to serovar autumnalis were higher than MAT titers to serovars pomona and grippotyphosa and persisted in some dogs for 16 weeks (6 weeks longer than for titers to serovar pomona). Of the 20 mature dogs, 13, 5, and 20 developed MAT titers of > 100 at 2 weeks to serovars pomona, grippotyphosa, and autumnalis, respectively. Titers to serovar pomona were higher and persisted in some dogs beyond 16 weeks after vaccination, compared with titers to serovars pomona and grippotyphosa, which persisted for 10 and 6 weeks, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: Subunit vaccines against serovars pomona and grippotyphosa induce MAT titers not only to homologous antigens but also to serovar autumnalis, which could lead to a misdiagnosis of leptospirosis caused by serovar autumnalis.     

A canine parainfluenza viral vaccine: immunogenicity and safety.

A canine parainfluenza viral vaccine was developed and shown to be safe by absence of clinical disease in vaccinated dogs and by inability to isolate vaccine virus from blood or nasopharyngeal swabs. Backpassage in susceptible dogs, using blood of vaccinated dogs, could not be demonstrated. The vaccine produced neutralizing antibody when administered either intramuscularly or subcutaneously; however, a significantly higher immune response was obtained by intramuscular inoculation. Differences in the antibody response were not produced by tenfold dilutions of vaccine virus ranging from 10(2.9) to 10(5.9) median tissue culture infective doses. The presence of neutralizing antibody was associated significantly with decreased respiratory shedding period of challenge virus by vaccinated dogs compared to seronegative control dogs. Six days after aerosol exposure to virulent challenge virus, 100% of the controls (n = 5) but only 15% of the vaccinated dogs (n = 3) shed virus. Seven days after challenge exposure, virus could not be recovered from the vaccinated dogs, but 80% of the control dogs shed virus. An anamnestic response occurred in vaccinated dogs but not in the seronegative control dogs following challenge exposure. A mild clinical disease was produced in 3 of the 5 seronegative control dogs but not in the 20 vaccinated dogs.