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New Zealand Veterinary Journal

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Post-vaccinal distemper encephalitis in two Border

Collie cross littermates
a b c d
RA Fairley , O Knesl , PA Pesavento & BC Elias
a
Gribbles Veterinary Pathology, PO Box 3866, Christchurch, New Zealand
b
Zoetis Animal Health, 5 Giralda Farms, Mailstop B1, Madison, New Jersey 07940, USA
c
Department Pathology, Microbiology and Immunology, School of Veterinary Medicine,
University of California-Davis, Davis CA 95616
d
Rangiora West Veterinary Clinic, 10 Oxford Rd, Rangiora, New Zealand
Accepted author version posted online: 14 Aug 2014.Published online: 19 Jan 2015.

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To cite this article: RA Fairley, O Knesl, PA Pesavento & BC Elias (2015) Post-vaccinal distemper encephalitis in two Border
Collie cross littermates, New Zealand Veterinary Journal, 63:2, 117-120, DOI: 10.1080/00480169.2014.955068

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 New Zealand Veterinary Journal 63(2), 117–120, 2015
Clinical Communication
Post-vaccinal distemper encephalitis in two Border Collie cross littermates
RA Fairley*§, O Knesl†, PA Pesavento‡ and BC Elias#
Abstract
CASE HISTORY: One 4.5-month-old male Border Collie cross
presented with aggression and seizures in October 2006. A 16-
month-old, female, spayed Border Collie cross presented with
hypersalivation and a dropped jaw and rapidly became
stuporous in September 2007. The dogs were littermates and
developed acute neurological signs 5 and 27 days, respectively,
after vaccination with different modified live vaccines
containing canine distemper virus.
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HISTOPATHOLOGICAL FINDINGS: Sections of brain in
both dogs showed evidence of encephalitis mainly centred on
the grey matter of brainstem nuclei, where there was extensive
and intense parenchymal and perivascular infiltration of
histiocytes and lymphocytes. Intra-nuclear and intra-
cytoplasmic inclusions typical of distemper were plentiful and
there was abundant labelling for canine distemper virus using
immunohistochemistry.
DIAGNOSIS: Post-vaccinal canine distemper.
CLINCIAL RELEVANCE: Post-vaccinal canine distemper has
mainly been attributed to virulent vaccine virus, but it may
also occur in dogs whose immunologic nature makes them
susceptible to disease induced by a modified-live vaccine virus
that is safe and protective for most dogs.
KEY WORDS: Dog, canine distemper, encephalitis, IHC, vaccine
Introduction
Post-vaccinal encephalitis has been recognised following the
administration of certain strains of attenuated canine distemper
virus vaccine but published cases are rare (Hartley 1974; Bestetti
et al. 1978; Cornwell et al. 1988; McCandlish et al. 1992; Mar-
tella et al. 2011). A reversion to virulence by the vaccine virus
has been proposed for some of these cases, and Appel (1978)
demonstrated reversion to virulence in the laboratory although
* Gribbles Veterinary Pathology, PO Box 3866, Christchurch, New Zealand
†
Zoetis Animal Health, 5 Giralda Farms, Mailstop B1, Madison, New Jersey
07940, USA
‡
Department Pathology, Microbiology and Immunology, School of Veterinary
Medicine, University of California-Davis, Davis CA 95616
#
Rangiora West Veterinary Clinic, 10 Oxford Rd, Rangiora, New Zealand
§
Author for correspondence. Email: rob.fairley@gribbles.co.nz
http://dx.doi.org/10.1080/00480169.2014.955068
© 2015 New Zealand Veterinary Association
117
the reversion in those dogs was not fatal and did not lead to
encephalitis.
Post-vaccinal distemper encephalitis was produced experimentally
in dogs given modified live vaccine containing distemper virus fol-
lowed by challenge with virulent parvovirus 3 days later (Kra-
kowka et al. 1982). The mechanism for the development of
encephalitis in these dogs was speculated to be due to immune
suppression from canine parvovirus infection.
This report describes the occurrence of post-vaccinal distemper
encephalitis, 1 year apart, in two littermates that received different
live vaccines. A congenital immune defect was a possible reason
for the development of the encephalitis.
Clinical history
Case 1
On 14 October 2006 a 4.5-month-old, male Border Collie cross
was presented with aggression and seizures; it was subject to
euthanasia 4 days later. The dog had been vaccinated with
Canvac 4 and Canvac BB (Pfizer Animal Health, Auckland,
NZ, now Zoetis Inc., Florham Park, NJ, USA) on 07 September
2006 and given a booster vaccination on 09 October 2006, 5 days
before its presentation. Canvac 4 contains live, attenuated strains
of canine parvovirus, canine distemper virus, canine adenovirus,
and canine parainfluenza virus, while Canvac BB is a killed,
vaccine for Bordetella bronchiseptica with adjuvant.
Case 2
On 10 September 2007 a 16-month-old, female, spayed Border
Collie cross was presented for veterinary examination with hyper-
salivation and a dropped jaw. Within 24 hours the dog was stu-
porous and it was subject to euthanasia on 12 September 2007.
On 14 August 2007, approximately 1 month prior to presen-
tation, the dog had received an annual booster with Vanguard
Plus 5 (Pfizer Animal Health) containing live attenuated parvo-
virus, canine distemper virus, canine adenovirus, and canine para-
influenza virus. The dog had been initially vaccinated 1 year
before on 18 July 2006 and booster vaccinated on the 10
August 2006 with Canvac 4 and Canvac BB.
Subsequent investigation revealed that these two dogs were litter-
mates owned by different families in the Rangiora area of North
Canterbury, New Zealand. The dogs had a mixed lineage includ-
ing Border Collie (dam of mother) and Airedale crossed with
perhaps Labrador or Great Dane (father of mother). The
mother of the dogs had been mated back to her father.
PBS Phosphate buffered saline
 118 New Zealand Veterinary Journal, 2015
Histopathological findings
Fresh brain tissue and brain fixed in 10% formalin was received
from each dog, and fixed heart, kidney, liver and lung from
Dog 1. There were no gross lesions. The fixed material was rou-
tinely processed and microscopic sections were cut at 4 µm and
stained with H&E.
Case 1
There were no lesions in the heart, kidney, liver or lung. In the
brain, lesions were present in the cerebral cortex, thalamus, mid-
brain, medulla and cerebellum. The cerebral cortex had
occasional, small, random foci of lymphocytes and microglial
cells in the white and grey matter. One focus had a few cells
with eosinophilic, cytoplasmic inclusions but most did not.
Occasional individual or clusters of four to six cysts resembling
Toxoplasma or Neospora spp. were present in the grey matter of
the cerebral cortex. Most of the cysts were separate from the
small, inflammatory foci; one cyst was present within an inflam-
matory focus.
The most severe lesions were in the grey matter of the thalamus
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where there was extensive and intense parenchymal and perivascu-
lar infiltration of histiocytes and lymphocytes. The grey matter in
the dorsal midbrain at the level of the rostral colliculus was infil-
trated with lymphocytes. The medulla had a few, small, focal areas
of lymphocytes and microglial cells in the grey matter, and the
cerebellum had several, small, focal areas of granular cell apoptosis
with vacuolation of the adjacent white matter and hypercellularity
of the adjacent molecular cell layer. Nuclear and cytoplasmic eosi-
nophilic inclusions were present in large numbers in neurons and
in lesser numbers of glial cells in the thalamus (Figure 1), and they
were also present in large numbers in the dorsal midbrain, and
within ependymal cells of the mesencephalic aqueduct contiguous
with an area of affected thalamic grey matter.
Case 2
There were no lesions in the cerebrum and lesions were present in
the cerebellum and in the brainstem from the rostral thalamus to
the medulla. The lesions in the brainstem mainly involved the
grey matter and the immediately adjacent white matter and
Figure 1. Light photomicrograph of a section of a brainstem nucleus,
from a 4.5-month-old dog, showing three neurons with large intranuc-
lear inclusions (arrows) and several intracytoplasmic inclusions (arrow-
heads) (H&E, bar = 10 µm).
Fairley et al.
consisted of parenchymal infiltrates of lymphocytes and histio-
cytes with perivascular lymphocytes. Eosinophilic nuclear and
cytoplasmic inclusions occurred in neurons and glial cells. The
inclusions were plentiful in some areas, and absent or few in
others. There were also scattered eosinophilic shrunken neurons
with pyknotic nuclei and many other pyknotic cells.
Immunohistochemistry
Immunohistochemistry was performed on samples from
both cases. Formalin-fixed, paraffin-embedded tissue sections,
that were 4 µm thick, were mounted on charged slides, and
air-dried overnight at 37°C. Sections were deparaffinized
through xylene to 100% reagent alcohol, and then treated with
0.3% hydrogen peroxide in 100% methanol for 30 minutes. Sec-
tions were rehydrated to water through 95% and 70% reagent
alcohols.
Antigen retrieval was performed with heat-induced epitope
retrieval for 30 minutes at 95°C, followed by a 20-minute
cool down. The retrieval solution was Target Retrieval Solution,
pH 6 (Dako Corp, Carpinteria, CA, USA). After antigen retrie-
val, slides were rinsed in deionised water and placed in 0.1 M
phosphate buffered saline (PBS), pH 7.4. Slides were then incu-
bated with blocking reagent (PBS with 0.02% Tween-20 and
10% normal horse serum) and blocked for 20 minutes. Mono-
clonal mouse anti-canine distemper virus (CMI 2–12; Custom
Monoclonals International, Sacramento, CA, USA) was diluted
at 1:200 in PBS with 0.02% Tween-20, and slides were incu-
bated for 1 hour at room temperature. Dako Envision
+System-HRP (Dako Corp), used according to the manufac-
turer’s instructions, was applied for 30 minutes to label mouse
anti- anti-canine distemper virus. Sections were counterstained
in Mayer’s haematoxylin and air dried. Canine cerebellum
from a case of natural distemper in California was used as a posi-
tive control. Negative controls were performed by using dupli-
cate sections in which isotype serum was used in place of the
primary antibody.
Figure 2. Light photomicrograph of a section of a brainstem nucleus, from
a 4.5-month-old dog, showing several neurons with intranuclear labelling
(arrows) and abundant intracytoplasmic labelling (arrowhead) with anti-
body to canine distemper virus (immunohistochemistry, bar = 25 µm).
 Fairley et al. New Zealand Veterinary Journal, 2015
Case 1
Immunohistochemistry was performed on blocks that included
thalamus, midbrain and medulla. There was marked cytoplasmic
and nuclear labelling with antibody to canine distemper virus of
neuronal cell bodies and their processes and of glial cell nuclei
in the grey matter of the thalamus (Figure 2), in cells in parts
of the ependyma of the thalamus, in the dorsal midbrain grey
matter, ventral midbrain meninges, and in occasional, scattered
cells in the white matter of the medulla.
Case 2
Immunohistochemistry was performed on a block that included
midbrain and cerebellum. There was marked cytoplasmic and
nuclear labelling with antibody to canine distemper virus of neur-
onal cell bodies and their processes and of glial cell nuclei in the
grey matter of the midbrain. There were scattered small areas of
labelling of cells and their processes in the cerebellar granular
and molecular cell layers. The nuclei and cytoplasm of cells in
parts of the meninges of the ventral midbrain also labelled
intensely.
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Discussion
The lesions and epidemiological features suggest that the encepha-
litis in these dogs was the result of infection with distemper vaccine
virus rather than being due to a field strain infection for several
reasons. First, canine distemper is an exceedingly rare disease in
New Zealand, assisted by the absence of wildlife reservoirs such
as raccoons (Procyon lotor), and over the last 30 years most prac-
titioners have never seen a case. The chances of two littermates
that had lived apart for over a year selectively developing natural dis-
temper seems remote especially as no other cases of distemper were
recognised in the dog population at the time and none have been
reported since then. Second, the encephalitis developed soon
after vaccination. Third, the lesions were similar to those described
in cases of post-vaccinal distemper by Hartley (1974). The lesions
were centred on the grey matter of mainly brainstem nuclei and
there was no demyelination. Alternative explanations, such as
natural infection when the dogs were still together as puppies,
with latent infection resulting in encephalitis in one of the dogs
over 12 months later, or vaccine-induced activation of latent infec-
tion, seem unlikely.
Although protozoal cysts were present in the cerebral cortex in
Case 1, they were interpreted to account for only a few small,
inflammatory foci in the cerebral cortex. Infection with Toxo-
plasma or Neospora spp. can occur concurrently with natural
canine distemper virus infection. No protozoa were detected in
Case 2 and as the protozoa were considered relatively unimportant
no attempt was made to determine whether they might be Toxo-
plasma or Neospora spp.
An intriguing difference between the two cases was the later devel-
opment of the disease in Case 2. This dog did not develop encepha-
litis after the initial course of vaccinations. The initial vaccinations
were the same as given to Dog 1, but the annual booster, following
which encephalitis developed, was with a different product.
The occurrence of disease in an individual is the result of the
outcome of the interaction between the agent, the host’s resistance,
and the host’s environment. In the present cases, the lack of reports
of post-vaccinal disease throughout the country, despite the wide-
spread use of these vaccines, and the occurrence of the disease in
119
two littermates vaccinated at different times, suggests that the ence-
phalitis was most likely due to a congenital susceptibility, rather
than a problem with the vaccine. A common environmental influ-
ence on the dogs’ immune systems is possible but unlikely given
that the dogs had lived in separate homes for over a year.
There are reports in the medical literature attributing immunode-
ficiency as the underlying reason for vaccine-induced disease. This
includes encephalitis after vaccination for the measles virus in a
child with profoundly low numbers of CD8 T lymphocytes
who was clinically normal and whose presumed immunodefi-
ciency went undetected (Bitnum et al. 1999). Cases of severe
primary immunodeficiency in children are usually detected
before vaccination, so vaccination of such children with live
vaccine is avoided, thus making the occurrence of post-vaccinal
measles encephalitis even rarer than might otherwise be the case
(Bitnum et al. 1999).
Hartley (1974) described two episodes of post-vaccinal distemper
encephalitis affecting large numbers of dogs in Australia in the late
1960s and early 1970s. The dogs had been vaccinated with a dis-
temper/hepatitis vaccine. Affected dogs were of six different
breeds, from 10 to 20 weeks old, and developed clinical signs
8–18 days post vaccination. These episodes were attributed to
an unusually virulent vaccine virus.
Although the histological lesions in the dogs in our report are
similar to those described by Hartley (1974) we believe the ence-
phalitis in our two dogs was likely due to a host susceptibility
(immunodeficiency) problem and not to a virulent vaccine virus.
The two multivalent vaccines given to these dogs have been
widely used in several countries without reported problems. The
vaccines contain different strains of live attenuated distemper
virus, Rockborn strain in Vanguard and CSL Masterseed B013
22/11/82 in Canvac, and are manufactured in different facilities.
The occurrence of fatal encephalitis may raise the issue of the
safety of live vaccines for some veterinarians. While distressing
for the owners of affected dogs, others would argue that these vac-
cines are highly efficacious and safe for the vast majority of dogs,
and without such good vaccines the animal welfare issues arising
from periodic distemper epidemics or even low-level endemic dis-
temper infection would be far worse.
Acknowledgments
Dr. Keith Thompson, Massey University took the
photomicrographs.
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