Zoonoses and Public Health

ORIGINAL ARTICLE

Meta-Analyses of Factors Associated with Leptospirosis in

Domestic Dogs

car-Aedo1 and G. Monti2
L. Azo
1
Graduate School, Faculty of Veterinary Sciences, Universidad Austral de Chile, Valdivia, Chile
2
Preventive Veterinary Medicine Department, Faculty of Veterinary Sciences, Universidad Austral de Chile, Valdivia, Chile

Impacts

• There are factors associated with leptospirosis in domestic dogs that coin-
cide in different studies worldwide.
• Two hundred and eighteen factors related with leptospirosis were identified
and 6 meta-analyses were performed to combine the results of observational
studies to obtain pooled measures as indicators of the infection risk.
• The variables ‘mixed-breed dogs’, ‘flooding occurrence in the habitat of the
dog’ and ‘working dogs’ were factors that increased the risk of leptospirosis,
while ‘being a dog less than 1 year old’ was a factor that decreased the risk,
but these associations were not statistically significant. Otherwise, the vari-
ables ‘male dogs’ and ‘urban dogs’ were statistically significant factors
increasing the risk of the disease.

Keywords:
Leptospirosis; domestic dogs; risk factors;
meta-analysis
Correspondence:

car-Aedo. Graduate School, Faculty of
L. Azo
Veterinary Sciences, Universidad Austral de
Chile, Campus Isla Teja, PO Box 567, Valdivia,
Chile. Tel/fax: +56 (63) 2 22 15 48;
E-mail: luciaazocaraedo@gmail.com
Received for publication March 5, 2015
doi: 10.1111/zph.12236
Summary
Factors related with leptospirosis in domestic dogs have been reported world-
wide. The aims of this study were to identify factors associated with this disease
described in different observational studies and to combine the coinciding factors
in at least four studies using meta-analyses, to obtain a pooled odds ratio (OR) as
measure of infection risk. A literature search was performed in electronic data-
bases, electronic databases of specific journals and search engines to find studies
published in English, Spanish and Portuguese available from January 1960 to Jan-
uary 2015. Two hundred and eighteen factors were identified in 31 publications
including cross-sectional and case–control studies. Finally, independent meta-
analyses were performed with six different variables, which included between 4
and 8 articles. The pooled OR indicated that the variables ‘mixed-breed dogs’,
‘flooding occurrence in the habitat of the dog’ and ‘working dogs’ were risk
factors for leptospirosis, while ‘being a dog less than 1 year old’ was a protective
factor; however, all these associations were not statistically significant. Otherwise,
the variables ‘male dog’ and ‘urban dog’ were statistically significant risk factors
for infection. This study highlights the need for more formal studies on the epi-
demiology of canine leptospirosis. Nevertheless, the study revealed that some risk
factors for infection coincided in different observational studies. These factors
could be considered to raise suspicion about the disease, especially when there is
a history of exposure to the bacteria.

The genus Leptospira is serologically classified into sero-

Introduction
Leptospirosis is a zoonotic disease of worldwide distribu-
tion, which is considered an important public health prob-
lem (Bharti et al., 2003; Chomel, 2014). It is caused by an
infection with a spirochaetal bacterium of the genus
Leptospira (Bharti et al., 2003; Goldstein, 2010).
vars, and it includes more than two hundred pathogenic
serovars, based on the structural heterogeneity of the bacte-
rial lipopolysaccharide (Xue et al., 2009). Serovars are
maintained in the environment by a variety of species such
as rodents, cattle and wildlife animals, which act as reser-
voirs of the bacterium in their kidneys (Greene et al.,

328 © 2015 Blackwell Verlag GmbH
Zoonoses and Public Health, 2016, 63, 328–336

car-Aedo and G. Monti
L. Azo
2008). The transmission of the infection occurs mainly by
the contact of intact mucous membranes or abraded skin
with urine or urine-contaminated soil, water or urine-con-
taminated food (Sykes et al., 2011).
Leptospirosis has a significant presence in canine clinical
practice with an important morbidity (Sessions and
Greene, 2004; Ananda et al., 2008). The seroprevalence of
canine leptospirosis varies according to the geographic
location. Some reported prevalences worldwide may range
from 1.8% in Australia (Zwijnenberg et al., 2008), 7.3% in
China (Shi et al., 2012), 17.1% in the USA (Davis et al.,
2008), 27.4% in Brazil (Bier et al., 2013), 29.4% in Italy
(Scanziani et al., 2002), 36.9% in Iran (Rad et al., 2004)
and 71.1% in India (Ambily et al., 2013).
Factors associated with canine leptospirosis have been
reported in a number of studies, for example, characteris-
tics of the dogs according to breed, gender, age, reproduc-
tive status, or lifestyle features such as contact with
livestock, rodents or wildlife species. Other studies focused
on associations between environmental variables and lep-
tospirosis such as rainfall and urban or rural localization of
the dogs (Ward et al., 2002, 2004a; Oliveira-Lavinsky et al.,
2012), but to date, no quantitative summary of the results
of observational studies describing factors associated with
the infection risk has been conducted.
A meta-analysis is a systematic approach to identify, syn-
thesize and combine the results of studies to obtain conclu-
sions about a specific topic or research question. It is an
analytical method where the results of independent and dif-
ferent studies are integrated and pooled in a common result
(Stroup et al., 2000; Leandro and Gallus, 2005). This
methodology has been frequently applied to randomize
controlled trials (Stroup et al., 2000); however, meta-ana-
lyses of observational studies are also becoming common
(Egger et al., 1998). Some examples of the application of
meta-analysis in observational studies exploring factors
associated with diseases and other topics are the publica-
tions authored by He et al. (1999), Renehan et al. (2004),
Sun et al. (2006), Taylor et al. (2014) and Falzon et al.
(2014).
The objectives of this study were (i) to identify factors
associated with leptospirosis in dogs described in different
observational studies and (ii) to combine those factors that
coincide in at least four studies using meta-analysis to
obtain a pooled measure of infection risk.
Materials and Methods
Literature search strategy
To find observational studies describing factors associated
with leptospirosis in dogs, a literature search was con-
ducted using the following electronic databases: Wiley-
Blackwell, EBSCOhost, Science Direct, Springer Link, Wiley
© 2015 Blackwell Verlag GmbH
Zoonoses and Public Health, 2016, 63, 328–336
Meta-Analyses on Leptospirosis in Dogs
Online, Pubmed Central and SCIELO, as well as electronic
databases of specific journals. In addition, the search was
performed on internet-based search engines including Goo-
gle, Google Scholar, Bing and Yahoo Search.
The references of the obtained articles were also
reviewed. The subsequent search of relevant documents
was performed on the electronic databases and search engi-
nes aforementioned.
The search was focused on documents written in English,
Spanish and Portuguese published from January 1960 until
January 2015. The keywords combinations used were (i)
English: canine leptospirosis, leptospirosis in dogs, lep-
tospirosis AND dogs OR canines, canine leptospirosis risk
factors and canine leptospirosis AND risk factors, (ii) Span-
ish: leptospirosis canina, leptospirosis en perros, leptospiro-
sis en caninos, factores de riesgo para leptospirosis canina y
factores de riesgo para leptospirosis en perros, (iii) Por-
tuguese: leptospirose canina, leptospirose em c~aes, fatores
de risco para leptospirose canina and fatores de risco para
leptospirose em c~aes.
Inclusion and exclusion criteria
The selected articles or documents included observational
studies on domestic dogs that described risk or protective
factors associated with leptospirosis and that pointed out
an indicator of the risk and their corresponding 95% confi-
dence interval (95% CI). Only original documents contain-
ing data concerning the association between animal,
lifestyle or environmental characteristics and the infection
were included. In addition, only studies with a clearly
defined diagnostic criteria depending on the diagnostic
tests used (criteria for considering the samples as positive
or negatives to Leptospira or cut-off values of antibody
levels in serologic tests) were considered.
Abstract of articles and other types of publications on
leptospirosis in dogs, such as literature reviews, evaluation
of diagnostic tests, letters to the editor and duplicated
articles were excluded.
Data extraction
The following data were extracted in each selected publica-
tion: author(s), year of publication, the geographic location
in which the study was carried out, study design and factors
related with leptospirosis with the measure of effect and
their 95% CI.
Other extracted data were sample size, number of posi-
tive animals in the diagnostic tests used, number of cases,
number of controls and number of individuals included in
the cohorts (exposed and unexposed groups), depending
on the study design (cross-sectional, case–control or cohort
studies).
329
Meta-Analyses on Leptospirosis in Dogs
Meta-analyses
In each selected publication, the factors potentially related
with leptospirosis were identified. Subsequently, the
repeated variables were grouped and independent meta-
analyses for those factors described in at least 4 different
publications were conducted.
The statistical heterogeneity between the selected studies
in each meta-analysis was determined using the Q statistic
approximation considering a significance level of 0.1. To
complement this, the inconsistency test (I2) was used with
values of 25%, 50% and 75% for low, moderate and high
heterogeneity, respectively (Leandro and Gallus, 2005).
When no evidence of heterogeneity was detected with
the tests aforementioned, a fixed effects model to obtain
the pooled measure of the effect was considered; otherwise,
a random effects model was used.
Forest plots were obtained to illustrate the estimated
measure of effect of each study and the overall pooled
effect.
To determine the presence of publication bias, the Begg0 s
and the Egger0 s tests were carried out considering a signifi-
cance level of 0.1. If there was evidence of publication bias
from any of these tests, the ‘trim and fill method’ was used
to estimate and correct for this publication bias.
All the analyses were performed using MIX Pro version
2.0 (Bax, 2011).
Evaluation of the methodological quality of the studies
included in the meta-analyses
After selection of the factors associated with leptospirosis
to be used in the different meta-analyses, an evaluation of
the methodological quality of the observational studies in
which the factors were described was conducted.
To evaluate cross-sectional studies, some characteristics
noted in the literature by VonElm et al. (2007) were taken
into account. These studies should indicate the following
features as indicators of good design: (i) the main elements
of the study design: with descriptions of the settings, loca-
tions and periods of recruitment, (ii) the sources and meth-
ods for the selection of the individuals included in the
study and the sampling strategy, (iii) clearly defined diag-
nostic criteria (for example, for the detection of a disease),
(iv) the number of outcome events or summary measures,
(v) a description of any effort to detect the potential source
of bias and 6) an indication of the statistical method(s)
used in the study.
To evaluate case–control studies, the following character-
istics stated by Frankena and Thrusfield (1997) and Wood-
ward (1999) were considered: (i) a definition of the
objectives of the study, (ii) a specification of the ‘case defi-
nition’ and clearly defined source population for cases and
330

car-Aedo and G. Monti
L. Azo
controls, (iii) a definition of the controls: which must be a
group of animals without the disease from the same source
population as the cases and with potential to develop the
disease during the study period, (iv) a description of any
effort to detect the potential source of bias, (v) a specifica-
tion of methods for disease determination (diagnostic tests
used) and (vi) a description of statistical method(s) used in
the study. In cohort studies, some features described by
Grimes and Schulz (2002), VonElm et al. (2007) and Song
and Chung (2010) were considered in the evaluation of the
methodological quality as follows: (i) clearly defined eligi-
bility criteria of the individuals included in the study, (ii)
the exposed and unexposed group should be selected from
the same population and it must have similar characteris-
tics, except for the exposure, (iii) the exposed and unex-
posed group must be at risk of developing the disease, (iv)
outcomes must be clear, specific and measurable, and it
must be comparable for the exposed and unexposed group
to avoid information bias, (v) report of the numbers of
outcome events or summary measures overtime, (vi)
description of methods for follow-up and minimizing attri-
tion and (vii) description the statistical method(s) used in
the study.
Results
After the literature search, 216 potential articles to be used
in the analysis were identified. In 12 of them, it was only
possible to obtain the abstract; therefore, they were
excluded, leaving a total of 204 full-text documents selected
for reviewing.
Of the 204 articles reviewed, 166 were excluded because
they did not describe factors associated with leptospirosis
or did not report the target data such as the measure of the
risk and the 95% CI. Consequently, 38 articles were
selected, but three studies were duplicates and two were
descriptions of clinical cases of leptospirosis in dogs; there-
fore, they were also excluded. Finally, 33 epidemiologic
studies met all the inclusion criteria, of which 22 (66.7%)
were written in English, 7 (21.2%) in Portuguese and 4
(12.1%) in Spanish. They were published from 1988 to
2015.
Regarding the study design, 19 of 33 articles (57.6%)
were cross-sectional and 14 (42.4%) were case–control
studies. Cohort studies were not found in the bibliographic
search.
Of the 33 articles that met the inclusion criteria, 31
described factors associated with leptospirosis using OR as
indicator of the risk. Two used relative risk and were
excluded from the analysis.
Two hundred and eighteen factors that describe the OR
as measure of the risk were identified in these 31 articles.
The factors associated with the disease that were reported
© 2015 Blackwell Verlag GmbH
Zoonoses and Public Health, 2016, 63, 328–336

car-Aedo and G. Monti
L. Azo Meta-Analyses on Leptospirosis in Dogs

in at least 4 different publications were as follows: ‘male publications. These variables were selected to execute six
dogs’, ‘mixed-breed dogs’, ‘dogs less than 1 year old’, independent meta-analyses.
‘working dogs’, ‘flooding occurrence in the habitat of the The meta-analyses performed included from four to
dogs’ and ‘urban dogs’, which appeared in 18 different eight studies (Fig. 1), which met the characteristics for

"Male dogs" "Mixed-breed dogs"

Aguiar et al (2007)
Ward et al
(2002)
Ward et al (2004a)
Batista et al
Ward et al
(2005)
(2002)
Ghneim et al (2003) Alton et al
Bañados (2009)
(1993)
Ward et al
Oliveira-Lavinsky et (2004a)
al (2012)
Martins et al (2013) Bañados
(1993)
Hennebelle
et al (2014)

0.0625 0.125 0.25 0.5 1 2 4 8 16 32 0.125 0.25 0.5 1 2 4 8 16

OR OR

"Dogs less than one year old" "Working dogs"

Ghneim et
Ghneim et al (2007)
al (2007)
Ward et al
Bañados (2004a)
(1993)
Ward et al
Zwijneberg (2002)
et al (2008)
Alton et al
Meeyam et (2009)
al (2006)
Oliveira- Hennebelle et al (2014)
Lavinsky et
al (2012)
Lee et al
Martins et (2014)
al (2013)

0.125 0.5 2 8 32 128 0.0625 0.125 0.25 0.5 1 2 4 8

OR OR

"Flooding occurrence" "Urban origin"

Ward et al
(2004b) Alton et al
(2009)

Raghavan et
Batista et al (2005)
al (2011)

Querino et
Querino et al (2003)
al (2003)
Lelu et al
(2015)
Raghavan et al (2011)

0.125 0.25 0.5 1 2 4 8 0.125 0.25 0.5 1 2 4 8 16 32

Fig. 1. Forest plots of the meta-analyses OR OR
performed.

© 2015 Blackwell Verlag GmbH
Zoonoses and Public Health, 2016, 63, 328–336 331
Meta-Analyses on Leptospirosis in Dogs
car-Aedo and G. Monti
L. Azo

cross-sectional and case–control studies with indicators of infection (OR < 1); however, these associations were not
a good study design. The studies included, in each meta- statistically significant (P > 0.05). Otherwise, being a ‘male
analyses, their designs and their geographic locations are dog’ (OR = 1.64; 95% CI = 1.38–1.95) and being an ‘ur-
listed in Table 1. ban dog’ (OR = 1.58; 95% CI = 1.04–2.39) were statisti-
Heterogeneity was detected in five of the meta-analysis cally significant risk factors for leptospirosis (P < 0.05)
performed; therefore, the random effects model was used (Table 2). Forest plots in Fig. 1 represent the OR of the
to determine the pooled OR. Only in the meta-analysis of studies included in each meta-analyses and the pooled OR
‘male dogs’, the fixed effect model was used because hetero- obtained.
geneity was not observed (Table 2). Only in the meta-analysis of ‘flooding occurrence in the
The pooled OR estimation in the meta-analyses of habitat of the dogs’ the Begg0 s and the Egger0 s tests showed
‘mixed-breed dogs’, ‘working dogs’ and ‘flooding occur- evidence of publication bias; however, the trim and fill cor-
rence in the habitat of the dogs’ showed that these variables rection indicated that no study needed to be added to cor-
were risk factors for leptospirosis (OR > 1) and ‘being a rect this bias (Table 2).
dog less than 1 year old’ was a protective factor for the
Discussion
Table 1. Data of the studies included in the meta-analyses performed
Leptospirosis is a disease with a complex epidemiology,
Meta-analysis/author(s) Study design Country and some determinants in their appearance are the interac-
Male dogs tion among the bacterium, reservoir hosts, susceptible ani-
Aguiar et al. (2007) Cross-sectional Brazil mals and the environment in which they all coexist (Lau
Ban~ados (1993) Case–control Chile et al., 2010). Several factors could also exert influence on
Ghneim et al. (2007) Case–control USA the presentation of the disease in domestic animals such as
Hennebelle et al. (2014) Case–control USA
climatic drivers (temperature, relative humidity), weather
Martins et al. (2013) Cross-sectional Brazil
events (increased rainfall, floodings and cyclones), hygienic
Oliveira-Lavinsky et al. (2012) Cross-sectional Brazil
Ward et al. (2002) Case–control USA and Canada conditions in the environment favouring rodent prolifera-
Ward et al. (2004a) Case–control USA tion, as well as the preventive measures taken, for example,
Mixed-breed dogs by animal health institutions, veterinarians and pet owners
Alton et al. (2009) Case–control Canada (Faine, 1994; Levett, 2001). In this study, we identify factors
Ban~ados (1993) Case–control Chile associated with leptospirosis in domestic dogs described in
Batista et al. (2005) Cross-sectional Brazil
different observational studies and combined them using
Ward et al. (2002) Case–control USA and Canada
meta-analysis to obtain pooled odds ratios as measure of
Ward et al. (2004a) Case–control USA
Dogs less than 1 year old the effect and as indication of infection risk.
Ban~ados (1993) Case–control Chile According Hooijmans et al. (2014), two studies as a
Ghneim et al. (2007) Case–control USA minimum are required to perform a meta-analysis and the
Martins et al. (2013) Cross-sectional Brazil methodological quality of the included studies is essential
Meeyam et al. (2006) Cross-sectional Thailand for the robustness of their results. In the bibliographic
Oliveira-Lavinsky et al. (2012) Cross-sectional Brazil
search in the present study, more than 200 factors related
Zwijnenberg et al. (2008) Cross-sectional Australia
with leptospirosis in domestic dogs were identified, which
Working dogs
Alton et al. (2009) Case–control Canada highlight the diversity of variables that researchers consid-
Ghneim et al. (2007) Case–control USA ered as potential risk or protective factors associated with
Hennebelle et al. (2014) Case–control USA the disease. However, it is notable that although some fac-
Lee et al. (2014) Case–control USA and Canada tors coincided between publications, these were not
Ward et al. (2004a) Case–control USA repeated in a large number of studies and consequently, 6
Ward et al. (2002) Case–control USA and Canada
meta-analyses were performed involving from four to eight
Flooding occurrence in the habitat of the dog
articles. All the studies included in the meta-analyses met
Batista et al. (2005) Cross-sectional Brazil
Querino et al. (2003) Cross-sectional Brazil the characteristics of good study design for cross-sectional
Raghavan et al. (2012) Case–control USA and case–control studies proposed by different authors
Ward et al. (2004b) Case–control USA (Frankena and Thrusfield, 1997; Woodward, 1999 and
Urban dogs VonElm et al., 2007); therefore, their results are reliable,
Alton et al. (2009) Case–control Canada even considering the limited number of studies.
Lelu et al. (2015) Cross-sectional Chile
Stroup et al. (2000) state that differences in designs and
Querino et al. (2003) Cross-sectional Brazil
outcomes are expected when observational studies are
Raghavan et al. (2011) Case–control USA
combined with meta-analysis, which contribute to the

332 © 2015 Blackwell Verlag GmbH
Zoonoses and Public Health, 2016, 63, 328–336

car-Aedo and G. Monti
L. Azo Meta-Analyses on Leptospirosis in Dogs

Table 2. Results of the meta-analyses performed including heterogeneity statistics, model used, pooled estimation, presence of publication bias and
trim and fill estimation

Heterogeneity statistics Pooled estimation

Meta-analyses Q test 2
I test Model OR 95% CI P value Publication bias Trim and fill estimation

‘Male dogs’ P = 0.58 I2 = 0% Fixed 1.64 1.38–1.95 <0.01* No N/A**

‘Mixed-breed dogs’ P = 0.007 I2 = 71.03% Random 1.24 0.79–1.94 0.34 No N/A
‘Dogs less than 1 year old’ P = 0.001 I2 = 86.71% Random 1.20 0.59–2.45 0.59 No N/A
‘Working dogs’ P = 0.001 I2 = 79.62% Random 0.97 0.52–1.83 0.94 No N/A
‘Flooding occurrence’ P = 0.005 I2 = 76.41% Random 2.00 0.88–4.55 0.09 Yes 0 studies
‘Urban dogs’ P = 0.003 I2 = 78.28 Random 2.23 1.01–4.94 0.04* No N/A

*Statistically significant.
**N/A (not applicable).

heterogeneity. For this, it is not surprising that heterogene-
ity was detected in five of six meta-analyses performed in
the present work. Other possible explanations to the
heterogeneity could be the variability in biologic character-
istics, such as age sex, or breed of the animals included in
the studies (Higgins et al., 2003; Hooijmans et al., 2014).
The causes of the heterogeneity in a meta-analysis can be
explored using meta-regression or subgroup analyses
(Moayyedi, 2004); however, these approaches were not car-
ried out considering the low number of available studies.
It is important to note that no cohort or prospective
studies about canine leptospirosis were included or even
found in the bibliographic search, which highlights the
need of these types of studies in the leptospirosis scientific
literature and to increase our understanding of the epi-
demiology of the infection in dogs as neither cross-sec-
tional or case–control studies can demonstrate causality
(Frankena and Thrusfield, 1997).
The diagnostic criteria used to classify the dogs with lep-
tospirosis (for example, different cut-off values in serology)
were employed as inclusion criteria and all the studies
included in this work met this criteria. However, not all the
reviewed studies stated if the vaccination status was consid-
ered to classify the animals as seropositive or seronegative.
In response to vaccination against Leptospira infection,
most of the dogs develop relatively low antibody titres
(from 1 : 100 to 1 : 400), which may persist at these levels
for 1–3 months after the application of the vaccine, while
other dogs develop high titres after vaccination, which
decline in longer times (Bolin, 1996). Therefore, timing of
vaccination could influence the results of serologic tests
and become a limiting factor in subsequent infection status
classification. This emphasizes that data about the time
since vaccination should be considered in the interpretation
of diagnostic tests such of MAT or ELISA.
The pooled odds ratios indicated that the variables
‘mixed-breed dogs’, ‘flooding occurrence in the habitat of
the dog’ and ‘working dogs’ were risk factors for lep-
tospirosis, while ‘being a dog less than 1 year old’ was a
protective factor, but these associations were not statisti-
cally significant. However, the fact that these variables were
described in more than three different scientific articles,
indicate that they should be considered as covariates in
further epidemiologic studies.
On the other hand, the variables ‘male dogs’ and ‘urban
dogs’ were statistically significant risk factors for lep-
tospirosis. The pooled OR revealed that male dogs have
1.64 greater odds of disease than females. Being a male dog
has been associated with higher frequency of leptospirosis
than females in surveys carried out in Argentina (Rubel
et al., 1997), Iran (Rad et al., 2004) and Trinidad (Ade-
siyun et al., 2006) and according Gaschen (2008), the
roaming behaviour of some male dogs could increase the
chances of infection, for example due to the possible con-
tact with stray dogs in cities, livestock and wildlife in rural
areas or environments contaminated with leptospires.
Dogs that live in urban areas showed 1.58 greater odds of
disease than rural canines.
Urban environments are important places where the
transmission of leptospirosis can be possible (Cachay and
Vinetz, 2005). Growing urbanization in suburban areas
promoting dog–wildlife contacts could allow the infection
with pathogenic serovars (Okewole and Ayoola, 2009). For
example, evidence of exposure to leptospires has been
found in peridomestic wildlife such as raccoons (serovar
Icterohaemorragiae), squirrels (serovars Grippotyphosa
and Canicola) and skunks (serovar Grippothyphosa) in
Conneticut, USA, which could be considered as potential
sources of Leptospira infection for domestic dogs in the area
(Richardson and Gauthier, 2004). Leptospires have also
been detected in kidney samples of rats (Rattus norvergicus)
captured in urban areas of Tokio, Japan (Koizumi et al.,
2009) and in Salvador, Brazil (Tucunduva de Faria et al.,
2008), which demonstrate the existence of reservoir hosts
for Leptospira in urban areas. The presence of leptospirosis
in urban settings could also be associated to demographic

© 2015 Blackwell Verlag GmbH
Zoonoses and Public Health, 2016, 63, 328–336 333
Meta-Analyses on Leptospirosis in Dogs
variations because an increment of human populations liv-
ing in urban slums (Picardeau, 2013), in which transmis-
sion of the leptospirosis among dogs could be facilitated by
environmental conditions such as overcrowding, poor
hygiene standards and inadequate sanitization (Burriel
et al., 2003).
Regarding publication bias, this was not observed in 5 of
the meta-analyses conducted. Publication bias can occur
when not all existing evidence about a particular topic is
properly represented in a meta-analysis (Higgins et al.,
2003). The lack of detection of this bias in the present study
could be explained because the bibliographic search was
broad and included scientific articles in three languages.
However, not all literature found could be reviewed (in 12
studies found, only the abstract was located) and it is possi-
ble that relevant articles have been missed. Moreover, in
the meta-analyses of ‘flooding occurrence in the habitat of
the dogs’, publication bias was detected and notably, the
trim and fill estimation showed that no studies should be
added to reduce the bias, which could be influenced by the
number of studies included (4 studies).
We are aware that the main limitation of this work is the
limited number of studies included in the 6 meta-analysis
performed; however, this the first approach to quantita-
tively summarize the results of available scientific evidence
in observational studies of leptospirosis in dogs. The study
revealed that there are factors associated with this disease
that coincide in different observational studies and that the
variables ‘male dogs’ and ‘urban dogs’ were statistically sig-
nificant factors that increase the risk of disease. As the
application of meta-analysis in animal health is a valuable
tool for improving health care (Hooijmans et al., 2014),
these results could be considered to raise suspicion about
the disease, especially when there is a history of exposure to
the bacteria.
Is important to note that other factors involving animal
characteristics, management practices by dog owners, or
environmental features apart from those found in the pre-
sent study could be associated with the disease, which
should be determined in further observational studies. In
addition, considering that canines can be important sen-
tinels for the occurrence of Leptospira infection, cohort
studies could be useful to establish more robust causal asso-
ciations.
Conflict of Interest
The authors declare that no conflict of interest exists.
Funding
L.Az

ocar-Aedo was supported by a scholarship from
CONICYT, Chile.
334

car-Aedo and G. Monti
L. Azo
References
Adesiyun, A., C. Hull-Jackson, N. Motoo, S. Halsall, R. Bennett,
N. Clarke, C. Whittington, and N. Seepersadsingh, 2006:
Sero-epidemiology of canine leptospirosis in Trinidad: sero-
vars, implications for vaccination and public health. J. Vet.
Med. 53, 91–99.
Aguiar, D., G. Cavalcante, M. Marvulo, J. Silva, A. Pinter, S.
Vasconcelos, Z. Morais, M. Labruna, I. Camargo, and
S. Gennari, 2007: Risk factors associated with anti-Leptospira
antibodies occurrence in dogs from Monte Negro County,
Rondonia, Brazilian Western Amazon. Arq. Bras. Med. Vet.
Zootec. 59, 70–76.
Alton, G., O. Berke, R. Reid- Smith, D. Ojkic, and J. Prescott,
2009: Increase in seroprevalence of canine leptospirosis and
its risk factors, Ontario 1998-2006. Can. J. Vet. Res. 73,
167–175.
Ambily, R., M. Mini, S. Joseph, S. Krishna, and G. Abhinay,
2013: Canine leptospirosis- a seroprevalence study from
Kerala, India. Vet. World 6, 42–44.
Ananda, K. J., T. Suryananarayana, P. Sharada, and P. D0 Souza,
2008: Diagnosis and treatment of leptospirosis in a dog. A
case report. Vet. World 1, 278–279.
Ba~
nados, R. 1993: Estudio retrospectivo de leptospirosis canina
diagnosticada en la cl
ınica veterinaria de peque~ nos animales
de la Universidad de Concepci
on, Chill
an (1987-1991).
Veterinarian Thesis, Universidad de Concepci
on, Chill
an,
Chile.
Batista, C., C. Alves, and S. Azevedo, 2005: Seroprevalence and
risk factors for leptospirosis in dogs from Campina Grande,
State of Para
ıba, Brazil. Arq. Bras. Med. Vet. Zootec. 57,
179–185.
Bax, L. 2011: MIX 2.0. Professional software for meta-analysis in
Excel. Version 2.0.1.4. BiostatXL California.
Bharti, A., J. Nally, J. Ricaldi, M. Matthias, M. Diaz, M. Lovett,
P. Levett, R. Gilman, M. Willig, E. Gotuzzo, and J. Vinetz,
2003: Leptospirosis: a zoonotic disease of global importance.
Lancet Infect. Dis. 3, 757–771.
Bier, D., S. Shimakura, V. Morikawa, L. Ullmann, M. Kikuti, H.
Langoni, A. Biondo, and M. Molento, 2013: Spatial analysis of
the risk of canine leptospirosis in the Vila Pantanal, Curitiba,
Paran
a, Brazil. Pesq. Vet. Bras. 33, 74–79.
Bolin, C., 1996: Diagnosis of canine leptospirosis: a reemerging
disease of companion animals. Semin. Vet. Med. Surg. 11,
166–171.
Burriel, A., C. Dalley, and M. Woodward, 2003: Prevalence of
Leptospira among farmed and domestic animals in Greece
Vet. Rec. 153, 146–148.
Cachay, E., J. Vinetz, 2005: A global research agenda for lep-
tospirosis. J. Postgrad. Med. 51, 174–178.
Chomel, B., 2014: Emerging and re-emerging zoonoses in dogs
and cats. Animals 4, 434–445.
Davis, M., F. Everman, C. Petersen, J. Vanderschalie, T. Besser, J.
Huckabee, J. Daniels, B. Hankock, M. Leslie, and R. Baer,
2008: Serological survey for antibodies to Leptospira in dogs
© 2015 Blackwell Verlag GmbH
Zoonoses and Public Health, 2016, 63, 328–336

car-Aedo and G. Monti
L. Azo
and raccoons in Washington state. Zoonoses Public Health 55,
436–442.
Egger, M., M. Schneider, and G. Smith, 1998: Meta-analysis of
observational studies BMJ 140, 1–8.
Faine, S., 1994: Leptospira and leptospirosis. CRS Press, Florida.
Falzon, L., T. O0 Neill, P. Menzies, A. Peregrine, A. Jones-Bitton,
J. vanLeeuwen, and A. Mederos, 2014: A systematic review
and meta-analysis of factors associated with anthelmintic
resistance in sheep. Prev. Vet. Med. 117, 388–402.
Frankena, K., and M. Thrusfield, 1997: Basics of observational
studies. In: Noordhuizen, J., K. Frankena, and der Van Hoofd
C. (eds), Application of quantitative methods in veterinary
epidemiology, pp. 101–134, Wageningen Pers, Wageningen.
Gaschen, F. 2008. Canine leptospirosis. Proceedings of the 33rd
World Small Animal Veterinary Congress, Dublin, Ireland.
2008. Available at: http://www.ivis.org/proceedings/wsava/
2008/lecture14/89.pdf?LA=1 (Accessed February 24, 2015).
Ghneim, G., J. Viers, B. Chomel, P. Kass, D. Descollonges, and
M. Johnson, 2007: Use of case-control study and geographic
information systems to determinate environmental and
demographic risk factors for canine leptospirosis. Vet. Res. 38,
37–50.
Goldstein, R., 2010: Canine leptospirosis. Vet. Clin. Small. Anim.
40, 1091–1101.
Greene, C., J. Sykes, C. Brown, and K. Hartmann 2008: Lep-
tospirosis. In: Greene, C. (ed), Enfermedades infecciosas del
perro y el gato, pp. 448–463. Interm
edica, Buenos Aires.
Grimes, D., and K. Schulz, 2002: Cohort studies: marching
towards outcomes. Lancet 359, 341–345.
He, J., S. Vupputuri, K. Allen, M. Prerost, J. Hughes, and P.
Whelton, 1999: Passive smoking and the risk of coronary
heart disease, a meta-analysis of epidemiologic studies. New
Engl. J. Med. 340, 920–926.
Hennebelle, J., J. Sykes, and J. Foley, 2014: Risk factors associ-
ated with leptospirosis in dogs from northern California:
2001-2010. Vector Borne Zoonotic Dis. 14, 733–739.
Higgins, J., S. Thompson, J. Deeks, and D. Altman, 2003:
Measuring inconsistency in meta-analyses. BMJ 227, 557–560.
Hooijmans, C., J. IntHout, M. Ritskes-Hoitinga, and M. Rovers,
2014: Meta-Analyses of animal studies: an introduction of a
valuable instrument to further improve healthcare. ILAR J. 55,
418–426.
Koizumi, N., M. Muto, T. Tanikawa, H. Mizutani, Y. Sohmura,
E. Hayashi, N. Akao, M. Hoshino, M. Kawabata, and H.
Watanabe, 2009: Human leptospirosis cases and the preva-
lence of rats harboring Leptospira interrogans in urban areas of
Tokyo, Japan. J. Med. Microbiol., 58, 1227–1230.
Lau, C., L. Smythe, S. Craig, and P. Weinstein, 2010: Climate
change, urbanization and leptospirosis: fuelling the fire?
Trans. R. Soc. Trop. Med. Hyg. 104, 631–638.
Leandro, G., G. Gallus. 2005: Meta-analysis in medical research.
Blackwell Publishing, Oxford.
Lee, H., L. Guptill, A. Johnson, and G. Moore, 2014: Signalment
changes in canine leptospirosis between 1970 and 2009 J. Vet.
Intern. Med. 28, 294–299.
© 2015 Blackwell Verlag GmbH
Zoonoses and Public Health, 2016, 63, 328–336
Meta-Analyses on Leptospirosis in Dogs
Lelu, M., C. Mu~ noz-Zanzi, B. Higgins, and R. Galloway, 2015:
Seroepidemiology of leptospirosis in dogs from rural and
slum communities of Los Rios Region, Chile. BMC Vet. Res.
11, 31. doi:10.1186/s12917-015-0341-9.
Levett, P., 2001: Leptospirosis Clin. Microbiol. Rev. 14, 296–326.
Martins, C., C. de Barros, C. Martins, M. Kikuti, L. Ullmann, R.
Pampuch, J. Hoffmann, H. Langoni, F. Ferreira, M. Molento,
and A. Biondo, 2013: Incidence of canine leptospirosis in the
metropolitan area of Curitiba, State of Paran
a, Southern Bra-
zil. Rev. Soc. Bras. Med. Trop. 46, 772–775.
Meeyam, T., P. Tablerk, B. Petchanok, D. Pichpol, and P.
Padungtod, 2006: Seroprevalence and risk factors associated
with leptospirosis in dogs Southeast. Asian J. Trop. Med.
Public Health 37, 148–153.
Moayyedi, P., 2004: Meta-analysis: can we mix apples and
oranges? Am. J. Gastroenterol. 99, 2297–2301.
Okewole, E., and M. Ayoola, 2009: Seroprevalence of leptospiral
serovars other than Canicola and Icterohaemorragiae in dogs
in Southwestern Nigeria. Vet. Archiv. 79, 87–96.
Oliveira-Lavinsky, M., R. Abou, G. Reuss, and H. Langoni, 2012:
Seroprevalence of anti-Leptospira spp antibodies in Bahia,
Brazil. Prev. Vet. Med. 106, 79–84.
Picardeau, M., 2013: Diagnosis and epidemiology of leptospiro-
sis. Med. Maladies. Infect. 43, 1–9.
Querino, A., A. Botazzo, R. Claret, F. Gibson, E. Eckehardt, R.
Lemos, and J. De Freitas, 2003: Risk factors associated to
leptospirosis in dogs in Londrina City-PR. Semina: Ciencias
Agrarias Londrina 24, 27–34.
Rad, M., A. Zeinali, J. Van Yousofi, and A. Tabatabayi,
2004: Seroprevalence and bacteriological study of canine
leptospirosis in Tehran and its suburban areas. Iran J. Vet.
Res. 5, 2–10.
Raghavan, R., Brenner, K., J. Higgins, D. Van der Merwe, and K.
Harkin, 2011: Evaluations of land cover risk factors for canine
leptospirosis: 94 cases (2002-2009). Prev Vet Med 101,
241–249.
Raghavan, R., K. Brenner, J. Higgins, J. Hutchinson, and K.
Harkin, 2012: Evaluations of hydrologic risk factors for canine
leptospirosis: 94 cases (2002-2009) Prev Vet Med 107,
105–109.
Renehan, A., M. Zwahlen, C. Minder, C. O0 Dwyer, S. Shalet, and
M. Egger, 2004: Insulin-like growth factor (IGF)-I,IGF bind-
ing protein-3, and cancer risk: systematic review and meta-
regression analysis. The Lancet 363, 1346–1353.
Richardson, D., and J. Gauthier, 2004: A Serosurvey of lep-
tospirosis in Connecticut peridomestic wildlife. Vector Borne
Zoon Dis: 3, 187–193.
Rubel, D., A. Seijo, B. Cerigoi, A. Viale, and C. Wisnivesky-Coli,
1997: Leptospira interrogans in a canine population of Greater
Buenos Aires: variables associated with seropositivity. Rev
Panam Salud P ublica 2, 102–105.
Scanziani, E., F. Origgi, A. Giusti, G. Lacchia, A. Vasino, G. Piro-
vano, P. Scarpa, and S. Tagliabue, 2002: Serological survey of
leptospiral infection in kennelled dogs in Italy. J. Small Anim.
Pract. 43, 154–157.
335
Meta-Analyses on Leptospirosis in Dogs
Sessions, J., and c. Greene, 2004: Canine leptospirosis: epidemi-
ology, pathogenesis and diagnosis. Compend Contin Ed Pract
Vet 26, 606–618.
Shi, D., M. Liu, S. Guo, S. Liao, M. Sun, J. Liu, L. Wang, Z.
Wang, S. Wang, D. Yang, and T. Chai, 2012: Serological sur-
vey of canine leptospirosis in southern China. Pak Vet J 32,
280–282.
Song, J., and K. Chung, 2010: Observational studies: cohort and
case-control studies. Plast Reconstruc Surg 126, 2234–2242.
Stroup, D., J. Berlin, S. Morton, I. Olkin, G. Williamson, D.
Rennie, D. Moher, T. Sipe, and S. Thacker, 2000: Meta-analy-
sis of observational studies in epidemiology A proposal for
reporting. JAMA 283, 2008–2012.
Sun, C. L., J. Yuam, W. Koh, and M. Yu, 2006: Green and black
tea and colorectal cancer risk: meta-analysis of epidemiologic
studies. Carcinogenesis 27, 1301–1309.
Sykes, J., K. Hartmann, K. Lunn, G. Moore, R. Stoddard, and R.
Goldstein:, 2011: 2010 ACVIM small animal consensus state-
ment on leptospirosis: diagnosis, epidemiology, treatment
and prevention. J. Vet. Intern. Med. 25, 1–13.
Taylor, L., A. Swerdfeger and G. Eslick, 2014: Vaccines are not
associated with autism: an evidence-based meta-analysis of
case control and cohort studies. Vaccine 32, 3623–3629.
Tucunduva de Faria T., M. Calderwood, M. Athanazio, A.
McBride, R. Hartskeerl, M. Pereira, A. Ko, and M. Reis, 2008:
Carriage of Leptospira interrogans among domestic rats from
336

car-Aedo and G. Monti
L. Azo
an urban setting highly endemic for leptospirosis in Brazil.
Acta Trop. 108, 1–5.
VonElm, E., D. Altman, M. Egger, S. Pocock, P. Gotzsche, and J.
Vandenbroucke, 2007: The Strengthening the Reporting of
Observational Studies in Epidemiology (STROBE) statement:
guidelines for reporting observational studies. Epidemiology
18, 800–804.
Ward, M., L. Glickman, and L. Guptill, 2002: Prevalence and risk
factors for leptospirosis among dogs in the United States and
Canada: 677 cases (1970-1998). J. Am. Vet. Med. Assoc. 220,
53–58.
Ward, M., L. Guptill, and C. Wu, 2004a: Evaluation of environ-
mental risk factors for leptospirosis in dogs: 36 cases (1997-
2002)J. Am. Vet. Med. Assoc. 225, 72–76.
Ward, M., L. Guptill, A. Prahl, and C. Wu, 2004b: Serovar-speci-
fic prevalence and risk factors for leptospirosis among dogs:
90 cases (1997-2002). J. Am. Vet. Med. Assoc. 22, 1958–1963.
Woodward, M., 1999: Epidemiology. Study design and data
analysis. Chapman & Hall/CRC, London.
Xue, F., J. Yan, and M. Picardeau, 2009: Evolution and patho-
genesis of Leptospira spp: lessons learned from the genome.
Microbes Infect. 11, 328–333.
Zwijnenberg, R., L. Smythe, M. Symonds, M. Dohnt, and
J. Toribio, 2008: Cross- sectional study of canine leptospirosis
in animal shelter populations in mainland Australia. Austr.
Vet. J. 86, 317–323.
© 2015 Blackwell Verlag GmbH
Zoonoses and Public Health, 2016, 63, 328–336