Serological Survey of Antibody to Neospora caninum in Japanese Dogs

Masumi SAWADA, Chun-Ho PARK1) , Hisayo KONDO1) , Takehito MORITA1) , Akinori SHIMADA1), Itsuro YAMANE2)
and Takashi UMEMURA
Laboratory of Comparative Pathology, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo 060–0818, 1)Department
of Veterinary Pathology, Faculty of Agriculture, Tottori University, Tottori 680–0945, and 2)Laboratory of Epidemiology, National
Institute of Animal Health, Ibaraki 305–0856, Japan
(Received 18 November 1997/Accepted 4 March 1998)

ABSTRACT. Prevalence of antibody to Neospora caninum (NC) in Japanese dogs were examined. The antibody was positive in 15 of 48
dogs (31.3%) reared in the dairy farms that had case of the abortions due to NC infection or had the cattle seropositive to NC, whereas the
prevalence was 7.1% (14 of 198 dogs) among the dogs kept in urban areas. In one dog breeder, all 17 Shetland sheepdogs older than 7
months were seropositive, and one pup was diagnosed as neosporosis 2 months before the first serological examination. The antibody
titers of the dogs kept at this breeder were almost unchanged for 1.5 years. Serological evidence of the dogs in the dairy farms and urban
areas indicates the transmission of NC between dogs and cattle. Also serological results of the dogs in one breeder may suggest the
potential horizontal transmission among dogs. — KEY WORDS: canine, Neospora, seroprevalence.
J. Vet. Med. Sci. 60(7): 853–854, 1998

Neospora caninum (NC) is an apicomplexan parasite that
can infect cattle, dogs, cats, rodents and other mammalian
animals [2]. NC is morphologically similar to Toxoplasma
gondii but natural transmission routes are not fully
understood except for its vertical transmission [2]. Canine
neosporosis has been reported from many countries
including Japan [2, 7]. Bovine neosporosis has also been
reported in Japan [5]. However, natural transmission of NC
between or among these species have not been demonstrated
yet. Also, serological prevalence of NC in Japanese dogs
have not been reported yet. Therefore, this study was
conducted to investigate the potential of natural transmission
of NC between dogs and cows and to obtain the serological
status of dogs in different environments.
The indirect fluorescent antibody test (IFAT) was used to
measure the antibody titers to NC. Tachyzoites of the JPA1
[8] and NC-1 (VMRD Inc., Wash., U.S.A.) isolates were
used as antigen for the IFAT, and a fluorescein
isothiocyanate labeled anti-dog IgG (BIOSYS) at a 1:100
dilution. There were no significant differences in the
reactions of these two isolates in the IFAT (data not shown).
Serum samples were heated and inactivated at 56°C for 30
min, and stored at −20°C until tested. All samples were
diluted in phosphate buffered saline (pH 7.4) and screened
at a dilution of 1:50. Positive sera were diluted up to
1:3,200. Antibody titers of ≧ 50 were considered to be
seropositive according to the supplier’s indication.
Sera were obtained from the following 4 groups.
Sera of urban dogs (Group 1): Sera of 198 dogs from 1
month to 17 years old were obtained from veterinary
practitioners in 28 prefectures. Of these samples, 122
samples previously were taken to measure the antibody titers
against viruses such as canine distemper virus, parvovirus
and canine hepatitis virus. The rest of the 76 samples were
obtained for the purpose of this study.
Sera of dogs kept in dairy farms (Group 2): Sera of 48
dogs reared in 11 dairy farms that had experienced bovine
neosporosis diagnosed by pathological examinations or had
raised cattle seropositive to NC were obtained from 9
prefectures in Japan. Ages of these dogs ranged from 5
months to 18 years old.
Sera of dogs from a Shetland sheepdog breeder (Group 3
and 4): One puppy, which were reared at one Shetland
sheepdog breeder, showed neurological disorders and were
diagnosed as neosporosis by pathological examinations (data
not reported). Serum samples from 20 dogs (2 months to 5
years old) reared at this breeder were taken as first sampling
(group 3). One and a half years later, serum samples from
19 dogs (1 to 11 years old) including 11 dogs previously
examined in the first sampling were obtained as second
sampling (Group 4).
The seroprevalence of NC were 7.1%, 31.3%, 85% and
100% for groups 1, 2, 3 and 4, respectively (Table 1). Titers
of the antibodies were from 1:50 to 1:400 in group 1, from
1:50 to 1:1,600 in group 2, from 1:50 to 1:1,600 in group 3
and from 1:50 to 1:800 in group 4 (Table 1). In groups 2, 3
and 4, seroprevalence between male and female were not
significantly different (chi-squared test P<0.05). In groups
1 and 3, seroprevalence in dogs under 1 year old were
significantly lower than dogs from 1 to 3 years old or from
4 to 6 years old (chi-squared test P<0.05). There were no
seropositive dogs younger than one year old in group 2, and
three pups of 2 months old were seronegative in group 3.
The relationships between seroprevalence and breed or
districts of the breeding in groups 1 and 2 could not be
examined due to the shortage of obtained information. In
group 4, all dogs including 8 newly examined adult ones
had NC antibody titers between 1:50 to 1:800. The antibody
titers of 11 dogs measured at both samplings (groups 3 and
4) were almost unchanged (the maximum change was 1
dilution).
Serological surveys for antibodies to NC of urban dogs
in Belgium and U.S.A. have shown seroprevalences of 11
and 2.2% in the IFAT (≧ 1/50 titer), respectively [1, 4].
The seroprevalence of dogs in England were reported to be
16.6 or 5.8% in the IFAT (≧ 1/50 titer) [3, 6]. In the
854 M. SAWADA ET AL.

Table 1. Neospora caninum IFAT titers in Japanese dogs

Number of dogs with IFAT titers Seroprevalence Total number of

<50 1:50 1:100 1:200 1:400 1:800 1:1600 (%) dogs examined

Group 1a) 184 8 1 4 1 0 0 7.1 198

Group 2b) 33 10 1 2 1 0 1 31.3 48
Group 3c) 3 9 2 1 1 1 3 85.0 20
Group 4d) 0 15 3 0 0 1 0 100.0 19

a) Urban dogs. b) Dogs kept in dairy farms. c) First sampling of Shetland sheepdog breeder’s dogs. d)
Second sampling of Shetland sheepdog breeder’s dogs.

present study, the seroprevalence of urban dogs in Japan
was 7.1% and this was at the average of those in other
countries [1, 3, 4, 6]. Transplacental transmission of NC is
the only confirmed natural transmission [2]. In the present
study, the prevalence of the dogs kept in dairy farms
suspected of having neosporosis were remarkably higher
than that of urban dogs (chi-squared test P<0.01). Based
on this serological evidence, we suspect that horizontal
transmission between cattle and dogs may be occurring in
this farm. There was a significantly higher seroprevalence
in dogs kept in the Shetland sheepdog breeder compared
with the dogs in groups 1 or 2 (chi-squared test P<0.01).
Furthermore, the dogs younger than 1 year old in group 2,
and 2-month-old pups in group 3 were seronegative and all
dogs older than 7-month-old in groups 3 and 4 were
seropositive, and some of these seropositive dogs had no
consanguinity relations with others. Therefore,
transplacental infection may not be the only way of
transmission and horizontal transmission among dogs may
be potential other transmission. We need further
investigations to examine the biological evidence of
horizontal transmission and identify the final host of NC
infection.
ACKNOWLEDGMENTS. We thank Dr. Takehisa Sohma,
Marupi Lifetech Co., Ltd. for supplying serum samples.
This study was financially supported in part by the Japanese
Ministry of Education, Science, Sports, and Culture (Grant-
in-Aid for Developmental Scientific Research, No.
07556070).
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