VECTOR-BORNE AND ZOONOTIC DISEASES

Volume 12, Number 1, 2012

ª Mary Ann Liebert, Inc.
DOI: 10.1089/vbz.2011.0684

Domestic Dogs (Canis familiaris) as Reservoir Hosts

for Rickettsia conorii

Michael L. Levin, Lindsay F. Killmaster, and Galina E. Zemtsova

Abstract

Rickettsia conorii is the causative agent of Mediterranean spotted fever (MSF) and Israeli spotted fever (ISF)
transmitted by the brown dog tick Rhipicephalus sanguineus. In areas where MSF or ISF are prevalent, dogs have
high prevalence of R. conorii -neutralizing antibodies. However, the true role of dogs in the persistence of the R.
conorii transmission cycle is unknown, and their reservoir competence for this pathogen has remained untested.
We assessed the ability of dogs infected with R. conorii to transmit the pathogen to previously uninfected Rh.
sanguineus ticks. Dogs were infected either via needle-inoculation of cultured rickettsiae or naturally via infected
tick bite. Dogs were monitored for clinical signs of infection, for rickettsemia by PCR, and for seroconversion and
were subjected to infestation with uninfected ticks at different time points. Rh. sanguineus larvae and nymphs
successfully acquired the agent from both needle-inoculated and tick-infected dogs and transmitted it transta-
dially. Tick-infected dogs remained infectious to ticks for at least a month postinfection. The molted ticks were,
in turn, infectious to naı̈ve dogs. These results demonstrate that dogs are capable of acquiring R. conorii from
infected Rh. sanguineus ticks and transmitting infection to cohorts of uninfected ticks, thus confirming for the first
time that dogs are indeed competent reservoirs for R. conorii. In addition, dogs with different genetic back-
grounds appear to differ in their susceptibility to R. conorii infection.

Key Words: Dog—Mediterranean spotted fever—Reservoir—Rickettsia conorii—Rhipicephalus sanguineus.

Introduction in epidemiology of human infections has not been conclu-

sively defined. Opinions on the matter differ to the extreme—

R ickettsia conorii is the causative agent of the

Mediterranean spotted fever (MSF) and Israeli spotted
fever. It is an obligately intracellular, slow-growing Gram-
negative bacterium. The disease is characterized by an acute
febrile illness with eschar at the location of the tick bite, re-
gional adenopathy, and disseminated maculopapular rash
including extremities. When treated with appropriate antibi-
otics, the disease usually has a benign course; however, about
6% of the cases are severe, and fatal cases sometimes occur
even in young, healthy adults (Raoult et al. 1986). The most
common vector of R. conorii is the brown dog tick, Rhipice-
phalus sanguineus. Although this tick can feed on a variety of
mammalian and even avian animals (Hoogstraal and Kaiser
1958, Paperna and Giladi 1974, Mumcuoglu et al. 1993, Szabó
et al. 2008), it is most closely associated with canines and
primarily with the domestic dog, Canis familiaris (Walker et al.
2000). For that reason alone, dogs stand to play an important
role in the epidemiology of MSF. However, the true role of the
dog in the maintenance of the agent’s natural circulation and
from being the most important reservoir (Rehacek 1993), to
merely carrying Rickettsia-infected ticks and in so doing
bringing the agent into the vicinity of humans (Espejo et al.
1993, Mumcuoglu et al. 1993).
Unfortunately, these different opinions are exclusively
based on indirect or circumstantial evidence. R. conorii has
been isolated from Rh. sanguineus ticks removed from dogs.
Serological studies conducted in various countries endemic
for MSF have demonstrated a correlation between human
disease and R. conorii antibody prevalence in canines (Tringali
et al. 1986, Keysary et al. 1988, Herrero et al. 1992). Moreover,
dog owners have higher odds of being infected with R. conorii
than persons without pets (Mumcuoglu et al. 1993, Bacellar
et al. 1995). In addition, several studies have reported the
presence of either viable rickettsiae or rickettsial DNA in the
dogs’ blood (Durand 1930, Senneville et al. 1991, Estrada-
Pena and Venzal Bianchi 2006, Solano-Gallego et al. 2006). In
one controlled laboratory experiment, Kelly et al. (1992) re-
peatedly reisolated the Zimbabwean strain of R. conorii from

Rickettsial Zoonoses Branch, Centers for Disease Control and Prevention, Atlanta, Georgia.

28
DOGS AS RESERVOIRS FOR Rickettsia conorii
the blood of artificially inoculated dogs and suggested that
dogs may be important in the epidemiology of this pathogen
by acting as domestic reservoirs of infection for ticks, albeit for
short periods. However, this supposition has remained un-
tested, as the authors did not attempt to infect ticks by feeding
them on infected dogs.
To date, there is no direct evidence of R. conorii transmis-
sion from infected dogs to uninfected ticks. Questions of
whether domestic dogs can amplify the infection in the pop-
ulation of vectors, or merely feed infected ticks and bring
them into contact with humans, remain unanswered. There-
fore, the goal of this study was to evaluate the reservoir
competence of dogs for R. conorii by assessing their ability to
acquire the pathogen and transmit it to previously uninfected
tick vectors. The first phase was to determine the ability of
dogs infected with R. conorii via needle inoculation to transmit
the agent to uninfected Rh. sanguineus ticks. This first phase
also served to supply ticks for the R. conorii-infected colony. In
the second phase of the study, we assessed the ability of the
dogs exposed to R. conorii via a tick bite to transmit the nat-
urally acquired infection to uninfected vectors.
Materials and Methods
Ticks and model animals
Rh. sanguineus ticks in this study were derived from our
colony of North American origin and maintained in our lab-
oratory as previously described (Troughton and Levin 2007;
Levin et al. 2009). Specific pathogen free New Zealand white
rabbits (Oryctolagus cuniculus) were used as hosts for feeding
all developmental stages of uninfected ticks. Between feed-
ings, ticks were kept in environmental incubators at
24C – 1C and 90% relative humidity. For the transmission
experiments, ticks were placed inside feeding bags glued to a
shaven area on a dog’s back. After engorgement, xenodiag-
nostic ticks were allowed to molt to the nymphal stage and
individually tested for the presence of rickettsial DNA.
For this study, we used 18–24-month-old male dogs–either
mixed breeds or purpose-bred beagles. All experimental
procedures were performed in accordance with Institutional
Animal Care and Use Committee approved protocols. All
dogs were housed indoors, in a climate-controlled animal
facility that precluded an unintended exposure to any ar-
thropod-borne agent including rickettsiae. The absence of
antibodies to spotted fever group rickettsiae in the blood of
each dog was confirmed before inoculation or infestation by
Table 1. Transmission of Two Strains of Rickettsia conorii from Inoculated Dogs
to Uninfected Rhipicephalus sanguineus Ticks
Xenodiagnostic ticks
Rickettsia strain Dog ID Day ticks placed Fed as
R.c. conorii A1 1 Larvae
(strain Malish) A2 2 Nymphs
B1 1 Larvae
R.c. israelensis B2 1 Larvae
(strain T-487) 2 Nymphs
B3 1 Larvae
2 Nymphs
29
the indirect fluorescent antibody (IFA) as described next. The
appetite, behavior, and level of activity of each dog were
monitored daily throughout the study. The weight and body
temperature of infected animals were measured thrice per
week. Venous blood and serum samples were collected twice
weekly after infection and tested for the presence of rickettsial
DNA and anti-R. conorii antibodies.
Dogs needle-inoculated with R. conorii
R. conorii isolates of African (R. conorii conorii strain Malish-
ATCC VR614) and Mediterranean (R. conorii israelensis strain
T487) origin were grown in Vero E6 cells at 32C in antibiotic-
free minimal essential medium supplemented with 2% fetal
calf serum and 2 mg/mL L-glutamine (CDC Core Facility).
Rickettsiae were purified by Renografin density gradient
centrifugation, frozen at - 70C in SPG with 5 mM MgCl2 and
1% Renografin76, and titered by plaque assay on E6 cells as
previously described (Eremeeva et al. 2003).
A mixed-breed dog A2 and a beagle A1 were intravenously
inoculated with 1 · 106 R. conorii conorii. Mixed-breed dogs B1
and B2, and a beagle B3 were intravenously inoculated with
1 · 106 R. conorii israelensis (ISTT). Rh. sanguineus larvae were
placed on dogs A1, B1, B2, and B3 on the day of inoculation,
and uninfected nymphs were placed on dogs A2, B2, and B3
one day after inoculation (Table 1). Engorged larvae and
nymphs were collected daily, allowed to molt, and used to
establish R. conorii-infected colonies, or for the subsequent
experiments involving tick-borne infectious challenge. Re-
presentative samples of freshly molted acquisition-fed ticks
were tested by PCR for the presence of rickettsial DNA.Dogs
were monitored daily for 4 weeks for changes in temperature,
weight, and behavior. Blood and serum samples were ob-
tained from all dogs three/week for 5–6 months for PCR and
serologic detection of rickettsial infection.
Dogs infected with R. conorii israelensis via tick bite
Ticks fed on dogs needle inoculated with R. conorii israe-
lensis were placed on mixed-breed dog C1 together with un-
infected Rh. sanguineus nymphs for proliferation of R. conorii
infection in the tick colony by cofeeding as previously de-
scribed (Zemtsova et al. 2010). Adult ticks derived from these
nymphs were used as the source of infection for the subse-
quent experiments. The dog C1 itself was not subjected to
follow-up xenodiagnostic infestations but was monitored for
Tested as Prevalence of infection % (infected/tested)
Nymphs 17 (25/150)
Adults 93 (37/40)
Nymphs 37 (37/100)
Nymphs 12 (3/25)
Adults 10 (2/20)
Nymphs 24 (6/25)
Adults 15 (3/20)
30
clinical signs of infection as well as by blood-PCR and
serology.
Ten male and 10 female Rh. sanguineus infected with R.
conorii israelensis by means of cofeeding on the dog C1 were
placed on naı̈ve beagles C2 and C3 (day 0) and allowed to feed
for 8 days. After removal of all adult ticks, dogs were fitted
with new feeding bags, and cohorts of uninfected Rh. san-
guineus larvae were placed on days 9, 16, 23, and 30 postin-
fection. To avoid inflammatory reactions at the feeding site,
the first and third groups of larvae were placed on dog C2,
and the second and fourth groups of larvae were placed on
dog C3 (Table 2).
Diminished survival of ticks exposed to the strain Malish
(Levin et al. 2009) precluded implementation of this second
phase using R. conorii conorii.
PCR and serology
Whole blood (200 lL) and serum (500 lL) samples from
dogs were aseptically collected into either 1.7 mL micro-
centrifuge tubes (Corning, Inc., Lowell, MA) containing 5 lL
of 2% EDTA (Sigma Aldrich, St. Louis, MO) or Microtainer
serum separator tubes (Becton Dickinson and Co., Franklin
Lakes, NJ). Blood samples were stored frozen at - 20C until
tested by PCR for the presence of R. conorii DNA, and sera
were refrigerated for serologic testing.
DNA extraction and PCR procedures were carried out in
separate facilities. DNA was extracted from ticks and blood
samples by using the Qiagen DNEasy Blood and Tissue kit
(Qiagen, Inc., Valencia, CA) according to manufacturer’s
protocols. The presence of rickettsial DNA was detected by
PCR using primers RR190-547F and RR190-701R to amplify a
154-bp fragment of the rompA gene of Rickettsia as described
by Eremeeva et al. (Eremeeva et al. 2003). Blood samples were
tested in duplicates with negative samples included in all
extraction and PCR rounds. Water was used as a no-template
negative control. PCR was interpreted as positive when both
duplicate reactions had positive results.
IFA was performed on dog sera, as previously described
(Lennette et al. 1995), using FITC labeled goat anti-dog IgG (c)
conjugate diluted as per manufacturer’s recommendations
(KPL, Inc., Gaithersburg, MD). Slides were spotted with
Rickettsia conorii Morocco (VR141) antigen, air-dried, and
fixed in acetone. Serum samples were initially screened at 1/
16 and 1/256 dilutions, and positive samples were titered to
the endpoint in a twofold dilution series. Serologic data are
reported as the reciprocal of the last dilution showing positive
fluorescence. Titers q16 were considered positive.
Table 2. Transmission of Rickettsia conorii
israelensis from Dogs Infected Via Tick Bite
to Uninfected Rhipicephalus sanguineus Larvae
Prevalence of infection in molted
Dog ID Day larvae placed nymphs % (infected/tested)
C2 9 12 (3/25)
C3 16 20 (5/25)
C2 23 20 (5/25)
C3 30 2 (1/50)
KILLMASTER ET AL.
Results
Transmission of R. conorii conorii and
R. conorii israelensis from needle-inoculated
dogs to Rh. sanguineus ticks
The dogs A1 (beagle) and A2 (mixed-breed) responded to
needle inoculation with R. conorii conorii with mild fever, loss
of appetite for 2–4 days, and transient lethargy during the first
week after inoculation. All symptoms spontaneously resolved
without intervention by the day 8 postinoculation. Rickettsial
DNA was continuously detectable in the blood of dog A1
from day 2 to 8 postinoculation, whereas dog A2 was blood-
PCR positive from day 3 though day 7 and again on day 24.
Dogs A1 and A2 seroconverted by day 10 and 7, and the
immune response peaked between 14 and 21 days postinoc-
ulation with antibody titers reaching 16,384 and 8192 re-
spectively. Both dogs remained blood-PCR negative and
seropositive for the remainder of observation with antibody
titers declining to 512 by day 152 postinoculation.
Rh. sanguineus larvae and nymphs fed on both R. conorii
conorii–inoculated dogs successfully acquired the infection
and transmitted it transstadially (Table 1). The resulting
prevalence of infection in nymphal ticks that had fed as lar-
vae on beagle A1 was 17%, and the prevalence of infection in
adult ticks that had fed as nymphs on the mixed-breed dog A2
was 93%.
Needle inoculation of dogs with R. conorii israelensis re-
sulted in variable clinical presentation. Dog B1 was febrile for
the first 5 days postinoculation, anorexic on days 2–4, and
lethargic on day 4. It was blood-PCR positive only on days 1
through 8 postinoculation, seroconverted by day 15, and ex-
hibited a peak antibody titer of 16,384. The antibody titers
declined to 512 by day 90 after inoculation and to 256 by day
156. Dog B2 experienced a decreased appetite but no fever,
and had detectable rickettsemia on days 4, 7, and 21 postin-
oculation. It seroconverted by day 7 and exhibited a peak
antibody titer of 4096 on days 14–18. The titers declined to 512
by day 92, and the dog became seronegative within 6 months
postinoculation. Conversely, the beagle-dog B3 did not ex-
hibit any symptoms after the inoculation; it ate and behaved
normally, and retained a normal body temperature. Blood
samples from B3 were all PCR negative for R. conorii. This dog
seroconverted by day 7, but the antibody titers reached only
132, and the dog became seronegative within only 3 months
postinoculation.
Rh. sanguineus larvae and nymphs fed on all three R. conorii
israelensis—inoculated dogs acquired the infection and
transmitted it transstadially (Table 1). The resultant preva-
lence of infection in freshly molted nymphs ranged from 12%
to 37%, and in adult ticks—from 10% to 15%. It is noteworthy
that dog B3 successfully transmitted infection to both xeno-
diagnostic larvae and nymphs, despite being PCR-negative
throughout the experiment.
Transmission of R. conorii israelensis from dogs
infected via tick bite to uninfected Rh. sanguineus
When mixed-breed-dog C1 was infested with ticks that had
previously fed on an R. conorii israelensis–inoculated dog, it
did not exhibit any symptoms of infection. However, rick-
ettsial DNA was detected in the dog’s blood on two occasions
(days 25 and 56 postinfestation), and antirickettsial antibodies
DOGS AS RESERVOIRS FOR Rickettsia conorii
developed by day 10. Antibody titers peaked at 8192 on days
14–21, declined to 512 by day 52, and the dog became sero-
negative at 227 days postinfestation. Blood samples for PCR
were collected thrice/week throughout the 7.5-month long
observation period, and only two samples were positive. It is
noteworthy that the immune response to rickettsial infection
had already passed its peak by the time the pathogen was first
detected by PCR, and antibody titers were on the decline
when rickettsial DNA was detected in the blood the second
time.
Beagle dog C2 exhibited decreased appetite on day 5
postinfestation and elevated temperature on days 4–6. Blood
was PCR positive on days 43, 45, and 48 postinfestation.
Seroconversion occurred by day 13, and antibody titer peaked
at 2048 on days 17–22. After that, antibody titers gradually
declined, and the dog became seronegative by day 214 post-
infestation. Here again, the antibody response was already on
the decline by the time of rickettsial DNA detection in the
dog’s blood. Beagle dog C3 showed no clinical signs of
R. conorii infection. Its appetite, temperature, and behavior
remained normal, and it failed to seroconvert. However, it
had a PCR-positive blood sample on day 43 postinfestation.
Uninfected Rh. sanguineus larvae were able to acquire
R. conorii from both dogs C2 and C3 and transmitted the agent
transstadially (Table 2). This transmission occurred at the time
when rickettsial DNA was not detectable in the blood by PCR,
and despite extremely high antibody titers in dog C2. The
dogs remained infectious to uninfected larvae for at least a
month after infected ticks had completed their feeding, al-
though the prevalence of infection varied between cohorts of
recipient ticks (Table 2). Larger proportions of ticks acquired
the pathogen if placed on dogs at two and 3 weeks after
placement of infected adult ticks than if placed at 1 or 4 weeks.
The difference in prevalence of infection was statistically
significant between the second or third cohorts and the fourth
cohort of ticks (Pv2 = 0.004).
Discussion
Results of this study demonstrate for the first time that dogs
infected with R. conorii conorii and R. conorii israelensis are
indeed infectious to Rh. sanguineus ticks. Moreover, we show
that dogs acquiring the R. conorii israelensis infection naturally,
via tick bite, can be infectious to ticks for at least 4 weeks after
initial exposure to the agent regardless of the presence or se-
verity of clinical symptoms. Successful transmission of
R. conorii israelensis from dogs to ticks was observed while
rickettsemia was undetectable by PCR, and in the presence of
high antibody titers.
In 1967, Burdgdorfer and Varma suggested that ticks serve
as both vectors and reservoirs for most of the spotted fever
group rickettsiae (Burgdorfer and Varma 1967). General ac-
ceptance of this hypothesis may simply be due to its frequent
reiteration, or stem from an assumption that vertebrate hosts
of ticks once exposed to a pathogen become immune for the
rest of their lives and, thus, are excluded from maintenance of
the natural transmission cycle. However, maintenance of SFG
rickettsiae with ticks as the only reservoir would require ef-
ficient transovarial and transstadial transmission with bacte-
ria causing no deleterious effect on the reproductive fitness or
viability of the tick host. This does not always appear to be the
case, as at least some strains of Rickettsia rickettsii and R. conorii
31
have been shown to negatively affect fecundity and survival
of infected ticks (Niebylski et al. 1999, Levin et al. 2009, So-
colovschi et al. 2009). This suggests that these agents could not
be maintained indefinitely without regular horizontal prolif-
eration among ticks through susceptible vertebrate hosts.
Since the principal vector of R. conorii—Rh. sanguineus—is a
common parasite of dogs worldwide, these hosts are rou-
tinely exposed to the pathogen and have a potential for af-
fecting its maintenance cycle—whether by increasing or
decreasing prevalence of infection in ticks. A number of
studies have demonstrated a correlation between human
disease and R. conorii antibody prevalence in canines (Tringali
et al. 1986, Keysary et al. 1988, Herrero et al. 1992). Also, dog
owners may have higher odds of being infected with R. conorii
than persons without pets (Mumcuoglu et al. 1993, Bacellar
et al. 1995).
In our study, all but one dog seroconverted within 2 weeks
postinfection, regardless of the mode of inoculation and de-
veloped high antibody titers. These titers, however, declined
significantly within the next 3 months. Two of our dogs that
seroconverted after exposure to infected ticks became nega-
tive after 214 and 227 days postinfection. Evidence for the
relatively short life of anti-R. conorii immunity in naturally
infected dogs can be found in a number of studies where both
the prevalence and titers of antibodies declined rapidly in
winter when ticks were inactive but rose again the next spring
(Espejo-Arenas et al. 1990, Espejo et al. 1993, Delgado and
Carmenes 1995, Ortuno et al. 2009). This is in contrast to the
serologic response in humans, who maintain IgG titers for
several years after contracting boutonneuse fever (Mansueto
et al. 1985). At least two surveys found no significant differ-
ences in seroprevalence to R. conorii between young (< 1 year
old) and old dogs (Delgado and Carmenes 1995, Harrus et al.
2007). This again confirms that canine antibodies lasted less
than a year.
Despite frequent exposure to R. conorii, dogs rarely become
clinically ill when infected by either infected ticks or inocu-
lation of infected blood (Durand 1930, Mumcuoglu et al. 1993,
Ortuno et al. 2009), or these symptoms are so mild and of such
short duration that they are not noticed. One report described
a febrile illness associated with R. conorii infection in three
Yorkshire terriers, which exhibited anorexia and lethargy for
2–3 days (Solano-Gallego et al. 2006). In our study, most of the
dogs had similar clinical symptoms of infection, although the
severity of symptoms varied depending on the bacterial
strain, the mode of infection, and on the genetic background
of the animals. Needle inoculation with the isolate Malish
caused mild fever, anorexia, and lethargy. These symptoms
appeared more pronounced in a mixed-breed dog than in a
pure-bred beagle. Needle inoculation with an identical dose of
R. conorii israelensis caused yet milder symptoms in mixed-
breed dogs and none at all in a beagle. When naı̈ve dogs were
subjected to infestation with R. conorii israelensis-infected
ticks, two out of three exhibited no adverse reaction at all.
Although a description of clinical symptoms after R. conorii
infection in dogs was not a subject of this study, our records
show that the propensity for clinical illness was lower in
beagles than in mixed-breed dogs.
Regardless of the symptoms, rickettsial DNA was repeat-
edly detected in the blood of most dogs whether needle in-
oculated or bitten by infected ticks. Interestingly, the dogs that
exhibited more pronounced clinical symptoms (A1, A2, and
32
B1) had only short periods of detectable rickettsemia lasting
less than 10 days postinfection. On the other hand, blood-PCR
was positive much later—between 25 and 56 days postinfec-
tion—in dogs with milder or no symptoms. It is also note-
worthy that in dogs infected with R. conorii israelensis via tick
bite, rickettsial DNA was detected much later than in needle-
inoculated dogs. This dissimilarity is likely due to the differ-
ence in the infectious dose received by animals, but may also
be related to the inoculation routes. Similarly to our results,
Estrada-Pena and Bianchi detected persistence of R. conorii
DNA in dog blood for at least 3 months (Estrada-Pena and
Venzal Bianchi 2006).
Uninfected ticks feeding on one of the naturally infected
dogs at the peak of antibody response successfully acquired R.
conorii and transmitted it transstadially. Further, in two of the
dogs, rickettsial DNA was detected well after this peak,
whereas antibody titers were on a decline. These episodes of
detectable rickettsemia did not trigger a boost in antibody
titers. Thus, the presence of antibodies per se did not prevent
dogs from being infectious to ticks, or from recurring rick-
ettsemia. The transmission of R. conorii despite the presence of
high antibody titers suggests that the antibodies may not be
protective, and indicates a possibility that even seropositive
dogs may serve as competent reservoirs upon reinfection.
Together, results of this study conclusively demonstrate for
the first time that the domestic dog (C. familiaris) is a compe-
tent reservoir of R. conorii. Dogs infected with R. conorii,
whether by artificial or natural means, were able to transmit
the infection to previously uninfected Rh. sanguineus ticks.
Dogs were infectious to ticks for at least a month after expo-
sure to infected ticks, and the transmission success was not
dependant on the presence of clinical symptoms. Further, the
highest rates of transmission were seen in the absence of de-
tectable rickettsemia in the blood, and in the presence of high
antibody titers. We subjected dogs to infestations with unin-
fected ticks for only up to 4 weeks after the initial exposure. It
remains to be studied whether the period of infectivity for
ticks in naturally infected dogs can last longer than 1 month
postinfection.
The mildness of a tick-borne infection in reservoir animals
normally would allow vertebrate hosts to remain active,
which will increase exposure of the animal to uninfected ticks,
and spreading the infected ones. Therefore, absence of severe
clinical illness in dogs infected with R. conorii via the natural
route of a tick bite indirectly confirms ecological coadaptation
between this agent and the domestic dog as a natural reser-
voir.
Relative ecological importance of the systemic transmission
(due to rickettsemia) in dogs vs. transmission between co-
feeding ticks remains to be assessed. Prevalence of infection in
ticks acquiring R. conorii israelensis from naturally infected
dogs in this study was lower than when donor and recipient
ticks were simultaneously fed (Zemtsova et al. 2010). How-
ever, the longevity of the systemic transmission may well
compensate for its lower efficiency and allow the infectious
dogs to produce more infected ticks and spread them over a
larger territory.
Although Rh. sanguineus in peridomestic environment
feeds primarily on dogs, it can also be found in a diverse range
of wild and domestic animals, including rodents, lago-
morphs, ungulates, carnivores, and humans (Mumcuoglu
et al. 1993, Walker et al. 2000, Kahn and Line 2010). Un-
KILLMASTER ET AL.
surprisingly, all these host species are also exposed to R.
conorii, and some of them may also serve as reservoirs
alongside or in competition with dogs. Antibodies against R.
conorii have been also detected in a variety of domestic ani-
mals including pigs, donkeys, cattle, goats, sheep, rabbits,
mules, and horses, potentially suggesting a role in mainte-
nance of MSF (Herrero-Herrero et al. 1989). The European
rabbit (O. cuniculus) has been suggested to play a role in
transmitting R. conorii along the French Mediterranean coast,
because a spectacular reduction in MSF cases was seen during
an outbreak of myxomatosis in 1952, which destroyed the
wild rabbit population (reviewed by Rovery and Raoult
2008). In addition, small ruminants have been suspected as
potential reservoirs of R. conorii based on the fact that in Cy-
prus the prevalence of infection in Rh. sanguineus ticks col-
lected from sheep and goats was significantly higher than in
those from dogs (Psaroulaki et al. 1999). The ability of these or
any other vertebrate hosts for transmission of R. conorii to
ticks following natural infection remains to be studied. Only
then will their relative importance as reservoirs in the main-
tenance of this agent be able to be compared with that of the
domestic dog.
Acknowledgments
The authors thank Davidia N. Grant for her invaluable help
with the maintenance of infected and uninfected tick colonies.
They gratefully acknowledge Dr. William L. Nicholson and
Aubree J. Roche for their contributions to this study.
Disclosure Statement
No competing financial interests exist.
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Address correspondence to:
Michael L. Levin
Rickettsial Zoonoses Branch
Centers for Disease Control and Prevention
1600 Clifton Road, MS G-13
Atlanta, GA 30333
E-mail: mlevin@cdc.gov