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Israel Journal of Veterinary Medicine  Vol. 70 (1)  March 2015 53 Neosporosis and Hepatozoonosis in Pups
INTRODUCTION
Neospora caninum, an apicomplexa protozoan from the family
Sarcocystidae is the causative agent of abortion in cattle and
neuromuscular diseases in dogs (1). Te defnitive hosts of
N. caninum are domestic dogs and wild canids. Dogs may be
infected by N. caninum horizontally by consumption or verti-
cally by in utero transmission of tachyzoites during pregnancy
(1, 2). In naturally infected dogs and cattle the predominant
route of infection is considered to be transplacental (1, 2).
Hepatozoon canis is an arthropod-borne infectious agent
of the apicomplexan protozoa from the family Hepatozoidae
in the suborder Adeleorin. H. canis transmission takes place by
ingestion by the intermediate host, the dog, of the brown dog
tick Rhipicephalus sanguineus, the defnitive host that contains
mature oocysts (3). Most domestic dogs probably become
infected with H. canis through grooming ticks from their
hair coat. Also like other apicomplexan parasites horizontal
transmission through the uterus from dam to its ofspring
has been demonstrated to take place (3).
Concurrent Neosporosis and Hepatozoonosis in a Litter of Pups
Mazuz, L.M.,
1, 2
Wolkomirsky, R.,
1
Sherman A.,
3
Savitzsky, I.,
1
Waner, T.,
4
Golenser,
J.
2
and Shkap, V.
1
1
Division of Parasitology, Kimron Veterinary Institute, 50250 Bet Dagan, Israel.
2
Department of Microbiology and Molecular Genetics, Te Kuvin Centre for the Study of Infectious and Tropical Diseases,
the Hebrew University of Jerusalem, Jerusalem, Israel.
3
HaMercaz HaVeterinary BaNegev, Rehov HaShomron 62, Beer Sheva, Israel.
4
Veterinary Clinic, 9 Meginay Hagalil Street, 76200 Rehovot, Israel.
*
Corresponding Author: Dr. Monica Mazuz, Division of Parasitology, Kimron Veterinary Institute, 50250 Bet Dagan, Israel. Email: monica@int.gov.il
ABSTRACT
Tis case report describes a concomitant infection in a litter of puppies with two apicomplexan protozoa
Neospora caninum and Hepatazoon canis. Te diferent potential routes of infection are discussed along with a
description of the course of the disease. A private practitioner in the South of Israel (Beer Sheva) submitted
blood samples for serological testing for toxoplasmosis and neosporosis from a pup of 6 weeks of age (from a
liter of 6 pups) showing neurological symptoms. Te results were seropostive for N. caninum and seronegative
for toxoplasmosis. After 3 weeks another pup also started to develop neurological signs. Samples including
blood smears, serum samples for serology for Neospora and Toxoplasma and fecal samples from all the pups.
Out of the 6 pups tested four were found seropositive for Neospora while all pups were seronegative for
Toxoplasma. Of the six pups, four were found to be infected with Heptaozoon canis, three of which were co-
infected with N. caninum. One pup was infected with only H. canis and another pup with only N. caninum and
a third pup was found to be negative for both H. canis and N. caninum. Treatment consisted of trimethoprim
sulfadiazine and clindamycin which were administered for six weeks. Te pups recovered completely, with
the exception of paresis of the left hind limb in the frst diagnosed puppy. An important conclusion from
this case report is the need to test all pups in a litter for N. caninum where even only one pup shows clinical
signs. Furthermore treatment at an early age and for a prolonged period of time appeared to be successful in
preventing the progression of the clinical signs. To the best knowledge of the authors this is the frst natural
concomitant infection case of N. caninum and H. canis in a litter of puppies.
Keywords: Neospora caninum; Hepatazoon canis; Concurrent Infection; Dog; Pups.
Israel Journal of Veterinary Medicine  Vol. 70 (1)  March 2015 Mazuz, L.M. 54
Concurrent infections in dogs are reported to occur with
a variety of pathogens (3). Often they may be transmitted
by a common vector such as the tick. However co-infecting
pathogens seemingly unrelated may be involved, resulting in
clinical manifestations which may make the clinical diagno-
sis more complex. Te relationship between the concurrent
infections in this case involves two diseases of puppies, neo-
sporosis and hepatozoonosis, both of which can be transmit-
ted in utero. On the other hand, infection by one agent may
infuence the susceptibility to a new infection which may
infuence the progression of the existing condition.
Tis case report describes a concomitant infection in a
litter of puppies with two apicomplexan protozoa N. cani-
num and H. canis. Te diferent potential routes of infection
are discussed along with a description of the course of the
disease. To the best knowledge of the authors this is the frst
natural concomitant infection case of N. caninum and H. canis
in a litter of puppies.
CASE REPORT
A sample of serum was received from a private practitioner
from a pup named “Fistuk” of 6 weeks of age from a liter of
6 pups born in the city of Beer Sheva in the South of Israel.
Te pup showed neurological symptoms which in the opinion
of the practitioner resembled either that of toxoplasmosis or
neosporosis. Te clinical signs in the pup were that of paresis
of the left hind limb which appeared to be progressing to
the right hind limb. In addition the pup appeared to be both
fecally and urinarily incontinent.
Te serum sample was tested by the immunofuorescence
antibody test (IFAT) for both Toxoplasma gondii and Neospora
caninum. A pronounced high titer for were found for N. cani-
num (1:12,800) while no reaction to T. gondii were observed.
Te pup was treated orally with trimethoprim sulfadiazine
(Resprim, Teva Medical, Petah Tikva, Israel) and clindamycin
(Dalacin C, Pfzer Pharmaceutical, Israel). Two weeks later
the pup showed an improvement in the right hind limb but
not in the left hind limb.
When the pups reached 9 weeks of age, another pup,
named Puf, also started to develop neurological signs of
paresis in the left hind limb and furthermore had difculty
in rising. Samples including blood smears, serum samples for
serology for Neospora and Toxoplasma and fecal samples were
collected from all the pups. Out of the 6 pups tested four
were found seropositive for Neospora with high antibody titers
(Table 1) while all pups were seronegative for Toxoplasma. Of
the six pups, four were found to be infected with Heptaozoon
canis, diagnosed from examination of their blood smears and
three of which were co-infected with N. caninum. One pup
was infected with only H. canis and another pup with only N.
caninum and a third pup was found to be negative for both
H. canis and N. caninum. No oocysts, or other parasites were
found in fecal samples.
Clinical signs resembling those of neosporosis were only
seen in the two pups described above. Te other two pups
with hepatozoonosis and/or neosporosis did not appear to
exhibit any detectable clinical signs of disease.
Positive pups for either N. caninum or H. canis parasites
were treated per os with trimethoprim sulfadiazine (15mg/kg
q12h) (Resprim, Teva, Petach Tikva, Israel) and clindamycin
(10mg/kg q12h) (Dalacin, Pfzer) for six weeks (2). All pups
were found to be healthy after 6 weeks after treatment. Te
incontinent pup Fistuk improved although the left hind limb
remained paralytic. Te pup named Puf recovered completely.
No H. canis parasites were observed in the blood smears
Table 1: Antibody titer for Neospora caninum and the detection of Hepatozoon canis gamonts in a litter of 6 pups
Date 10-Oct-12 5-Nov-12 11-June-13
Name
Ab titers
N. caninum
Ab titers
N. caninum
Blood smear
Ab titers
N. caninum
Blood smear
Fanta n.d. 1:12800 Hepatozoon 1:12800 Negative
Pupa n.d. 1:12800 Negative 1:12800 Negative
Lipstick n.d. Negative Hepatozoon Not done Negative
Popcorn n.d. Negative Negative Not done Negative
Puf n.d. 1:12800 Hepatozoon 1:12800 Negative
Fistuk 1:12800 1:12800 Hepatozoon 1:12800 Negative
n.d. = Not done
Case Reports
Israel Journal of Veterinary Medicine  Vol. 70 (1)  March 2015 55 Neosporosis and Hepatozoonosis in Pups
examined at the end of the six week treatment period. Te
four Neospora seropositive puppies continued with high
antibody titers.
DISCUSSION
Te predominant route of natural infection in dogs by
Neospora caninum is considered to be transplacental (1, 2). Te
bitch from this report from the south of Israel was adopted
by the current owner as a stray dog. It is suggested that free
roaming dogs in urban areas have a greater seroprevalence
for N. caninum than companion pets (4).
It has been reported that a variable number, but not all
pups in a litter from a dam infected with N. caninum may
have clinical manifestations (4, 5, 11). Indeed, in this litter
four of the six pups presented as infected, judged by sero-
logical investigation (IFAT) and of them only two showed
clinical signs at diferent intervals after birth. An important
conclusion from this study is the need to test all puppies in a
litter for N. caninum where even only one pup shows clinical
signs. From the frequency of the disease among the pups in
this litter this conclusion appears to be applicable to both N.
caninum and H. canis.
Te clinical signs presented by the pups were typical of
neosporosis for their age: Congenitally infected pups tends
to exhibit a more severe form of the disease (2). Pups of less
than 6 months of age exhibit a severe disseminated form
showing ascending paralysis of the limbs with the pelvic
limbs afected more severely than the thoracic limbs (1, 6).
Muscle atrophy, contracture and fbrosis occur in many pups
(2). Gradual, progressive, ascending paralysis with hyperex-
tension of the hind limbs in congenitally infected puppies
is the most common clinical manifestation of the disease in
dogs less than 6 months of age (2).
H. canis infection causes disease ranging in severity from
an incidental hematologic fnding in an apparently healthy
dog to a debilitating and life threatening illness (3). Even so,
when high parasitemia is present H. canis may also induce
severe clinical manifestations (e.g. lethargy, fever, anorexia,
weight loss, lymphadenomegaly and anemia) associated with
a high parasite load (3, 7). Te possibility that the clinical
signs were exacerbated due to the concurrent infection with
H. canis cannot be completely discounted, however H. canis
is found primarily in the hemolymphatic tissues and not
directly related to the neurological system (3).
Concurrent infections with H. canis in dogs have been
described with Ehrlichia canis and Babesia canis where all
three infections are transmitted by the same tick vector,
R. sanguineus. Other pathogens reported to be involved in
concurrent infections include parvovirus, canine distemper,
Anaplasma phagocytophilum, Anaplasma platys, Toxoplasma
gondii (8) and Leishmania infantum. To the best knowledge
of the authors’ coinfection of Heptozoon canis with Neospora
caninum has not been previously described.
Te seroprevalence of H. canis in dogs in Israel has been
reported to be 33% (9). Te incidence of clinical disease for
H. canis is considered to be much lower than the exposure
rate with most dogs probably undergoing a subclinical form
of the disease. In fact in the litter described in this study one
pup was infected with H. canis only and was not reported to
show clinical signs. Although the route of infection cannot
be defnitely ascertained, it seems likely that the pups were
infected in utero. Horizontal transmission through in the
uterus from the dam to its ofspring has been demonstrated
in H. canis. In a study of naturally infected pregnant bitches
meronts were found in the spleen of a pup that died 16 days
after birth and blood gamonts were detectable as early as 21
days in other pups. Te possibility of these pups being natu-
rally infected by ingestion of the Rhipicepahalus sanguineus
ticks cannot be completely discounted, however taking into
account the young age of the pups and the relatively high rate
of infection within the litter this seems unlikely. Furthermore
it would have been necessary for the young pups to ingest the
ticks and this makes the possibility of infection after birth as
even more unlikely.
Symptoms seen in the two pups were strongly sug-
gestive of infection with N. caninum (1). Treatment with
trimethoprim sulfadiazine and clindamycin for six weeks
was successful in treating the two afects pups, one pup had
recovered completely, and the other remained with paresis of
the left hind limb whereas the other signs including urinary
and fecal incontinenence were alleviated. Although the cur-
rent treatment protocol for H. canis is imidocarb dipropionate
alone or in combination with doxycycline, a similar treatment
used in this study has been recommended for several years
in the treatment of H. canis (10). Tis treatment regime was
not applied in the treatment of two puppies in this study
indicating that H. canis may not have been involved in the
symptomology of this concurrent infectious condition.
Te exact interaction of N. caninum and H. canis in the
Case Reports
Israel Journal of Veterinary Medicine  Vol. 70 (1)  March 2015 Mazuz, L.M. 56
pathogenesis of the symptoms is unknown but from the
treatment outcome it does appear that N. caninum did play
a prominent or maybe even a singular role.
In conclusion, this case study is to the best knowledge
of the authors the frst report of a concomitant infection
of neosporosis and hepatozoononosis in a litter of puppies.
Treatment at an early age and for a prolonged period of time
appeared to be successful in preventing the progression of the
clinical signs and possibly preventing clinical signs in other
litter mates which were infected. Our study is in agreement
with others who shown that in naturally infected dogs, treat-
ment can improve clinical signs but may not eliminate the
infection, as the puppies remained seropositive (11).
Te clinical signs were strongly indicative of those caused
by N. caninum although the role of H. canis in the develop-
ment of the symptomology could not be established.
ACKNOWLEDGMENT:
Appreciation to the owner of the puppies Ronit, for her cooperation
and dedication.
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Case Reports