Article

Disease investigations for equine infectious anemia in Canada

(2009–2012) — Retrospective evaluation and risk factor analysis
Katharina L. Lohmann, Carolyn R. James, Sara N. Higgins, Krista J. Howden, Tasha Epp

Abstract — This retrospective study describes the detection of equine infectious anemia (EIA) during Canadian
Food Inspection Agency (CFIA) disease investigations in Canada, examines aspects of importance for disease
control, and evaluates potential animal-level risk factors for EIA in high-risk horses. Based on review of all
EIA-positive samples and all samples collected during disease investigations (N = 4553) over a 4-year period (2009
to 2012), 409 EIA cases were detected. Horse owners with EIA cases owned between 1 and 60 affected animals,
and 49 horses seroconverted during a disease investigation period. Twenty-nine percent of cases (n = 68) for which
this information was available had, or possibly had, clinical signs of EIA. Using a mixed effects logistic regression
model, horses in older age groups were at greater odds of having a positive EIA status. The study emphasizes the
importance of disease investigation activities when EIA is detected and identifies age as an animal-level risk factor
in high-risk horses.

Résumé — Enquêtes médicales pour l’anémie infectieuse équine au Canada (2009-2012)  –  Évaluation
rétrospective et analyse des facteurs de risques. Cette étude rétrospective décrit la détection de l’anémie infectieuse
équine (EIA) durant les enquêtes médicales de l’Agence canadienne d’inspection des aliments (CFIA) au Canada,
examine les aspects importants pour la maitrise de la maladie, et évalue les facteurs de risque potentiels au niveau
des animaux pour l’EIA chez les chevaux à risque élevé. Sur la base d’une revue de tous les échantillons positifs
pour l’EIA et tous les échantillons prélevés durant les enquêtes (N = 4553) pendant une période de 4 ans
(2009-2012), 409 cas d’EIA furent détectés. Les propriétaires de chevaux avec EIA possédaient entre
1 et 60 animaux affectés, et 49 chevaux ont séro-converti durant une période d’enquête. Vingt-neuf pourcents des
cas (n = 68) pour lesquels l’information était disponible avaient, ou avaient possiblement eu, des signes cliniques
d’EIA. Utilisant un modèle de régression logistique à effets mixtes, les chevaux des groupes d’animaux plus âgés
étaient à plus grand risque d’avoir un statut positif pour l’EIA. Cette étude fait ressortir l’importance des activités
d’enquêtes médicales lorsque l’EIA est détectée et identifie l’âge comme étant un facteur de risque au niveau de
l’animal chez les chevaux à risque élevé.
(Traduit par Dr Serge Messier)
Can Vet J 2019;60:1199–1206

Introduction EIA virus, and in the absence of licensed vaccines or effective

E quine infectious anemia (EIA) is a reportable disease in

Canada and a national disease control program is admin-
istered by the Canadian Food Inspection Agency (CFIA) under
the Health of Animals Act and Regulations (1,2). Infected horses
(subsequently referred to as all equines) remain carriers of the
treatments (3), disease control is accomplished by removal of
infected horses from the population through euthanasia or
life-long quarantine. Blood samples for owner-requested vol-
untary surveillance testing for EIA are collected by accredited
veterinarians and tested at private or provincial laboratories

Department of Large Animal Clinical Sciences, Western College of Veterinary Medicine, University of Saskatchewan, 52 Campus
Drive, Saskatoon, Saskatchewan S7N 5B4 (Lohmann, Epp, Higgins); Terrestrial Animal Health Epidemiology and Surveillance
Section, Canadian Food Inspection Agency, 8403 Coronet Road NW, Edmonton, Alberta T6E 4N7 (Howden); Canadian Food
Inspection Agency, 59 Camelot Drive, Ottawa, Ontario K1A 0Y9 (James).
Address all correspondence to Dr. Katharina L. Lohmann; e-mail: k.lohmann@usask.ca
Dr. Higgins’ current address is APHA, Melcombe Court, 1 Cumberland Drive, Weymouth, DT4 9TT, United Kingdom.
Dr. Howden’s current address is One Health Scientific Solutions, 150 Chippewa Road, Unit 258, Sherwood Park, Alberta T8H 0K4.
Use of this article is limited to a single copy for personal study. Anyone interested in obtaining reprints should contact the CVMA
office (hbroughton@cvma-acmv.org) for additional copies or permission to use this material elsewhere.

CVJ / VOL 60 / NOVEMBER 2019 1199

 CFIA-approved laboratory CFIA National Reference Laboratory

cELISA cELISA AGID EIA pos

EIA neg EIA neg EIA neg

A R T I C LE

Accredited veterinary CFIA veterinary inspectors

practitioners

Figure 1.  Overview of the protocol for equine infectious anemia (EIA) testing in
Canada. Solid arrows represent a non-negative (cELISA) or positive (AGID) result,
and dashed arrows represent a negative test result. For the few samples that have
non-definitive AGID results, further testing, and possibly sampling, is performed as
per established protocol. cELISA — competitive enzyme-linked immunosorbent assay;
AGID — agar-gel immunodiffusion; CFIA — Canadian Food Inspection Agency; EIA
neg — negative for EIA; EIA pos — positive for EIA (EIA case).

approved for EIA screening by the CFIA. Samples with non-
negative test results during this screening process (laboratory
referrals, Figure 1), and blood samples collected by CFIA
inspectors during disease investigations and before export of
horses to countries other than the US and Mexico, are tested at
the CFIA National Reference Laboratory to confirm a horse’s
EIA status. Less intense owner-requested surveillance for EIA
in the western provinces was associated with an increase in the
detection of EIA cases (2).
Complementary to voluntary surveillance efforts, the current
EIA control program in Canada mandates disease investigation
activities when an outbreak of EIA is detected. Based on OIE
definitions, an outbreak of EIA is the occurrence of 1 or more
confirmed positive cases in an epidemiological unit. An epi-
demiological unit comprises 1 or more horses that are located
on a premises where the opportunity for disease transmission
exists via the exchange of blood or bodily fluids. This may occur
through various events including the activity of large biting flies
(primarily tabanids), the use of contaminated medical or surgi-
cal equipment, biting among horses, breeding, pregnancy, and
nursing (3–5). Disease investigation activities undertaken by the
CFIA include “sampling, performing epidemiological investiga-
tions, imposing movement restrictions, monitoring [and] order-
ing destruction” (6). Disease investigation testing pertains to all
horses that have been “in contact with the positive animal within
30 d of the sampling date” (7). This may include horses on the
same premises as the infected horse (index premises), horses on
a different premises in close proximity to the index premises
(i.e., fence line contact), and horses that had contact with the
infected horse at some point during the previous 30 d but are
no longer on the same premises (e.g., exposure during an event,
hospitalization, temporary housing). The policy on the timing
of testing in-contact horses takes into account the magnitude of
risk for virus transmission, which is thought to increase during
fly-biting season and with the presence of clinical signs in the
positive horse or its on-premises contacts as these horses likely
have significantly higher viral loads (8). The policy also accounts
for the fact that although sensitive assays detect antibodies
against the EIA virus as early as 14 to 28 d after infection (3,9),
a small proportion of infected horses may, for various reasons,
take longer to seroconvert (3,10). At a minimum, in-contact
horses are therefore sampled 45 d after the last known exposure
to an EIA case to allow for detectable levels of antibody to be
present. All horses on the index premises are tested until they
receive a negative test result.
One aspect in the assessment of risk for EIA virus transmis-
sion concerns housing distance between horses. Transmission of
EIA virus by tabanids is purely mechanical (11) and requires that
infected blood is transmitted to a second susceptible host within
hours (3,12), meaning it only takes place when a blood meal
is interrupted and then completed on a second host. Although
tabanids can travel long distances in general, they are unlikely
to change hosts when these horses are more than 50 m apart
(13–15), and disease investigation testing includes horses that
reside on a premises within 200 m of the index case. Physical
distance between horses may be less relevant in the context of
iatrogenic transmission.
In addition to distance, several previous studies have
addressed environmental and individual risk factors for natural
transmission of EIA virus. In 1 study (16), horse flies were most
active on hot days with bright sunshine and little or no wind,
while another study reported that tabanid activity was affected
by barometric pressure, temperature, relative humidity, time
of day, and cloud cover (17). Young horses, especially young
foals, were at reduced risk of infection (18), which is possibly
attributable to a more exaggerated defensive response to tabanid
feeding. Horses residing in flooded areas were at higher risk of
being seropositive in Brazil (19). Interestingly, several studies
showed that tabanids are less attracted to light-colored than
dark-colored animals, and also less attracted to certain striped
and spotted patterns than to homogenous colors (20,21). This
behavior is attributable to differences in polarized light reflection

1200 CVJ / VOL 60 / NOVEMBER 2019

based on coat color; however, to our knowledge, the relevance
of these findings has not been evaluated in the context of EIA
transmission.
The study herein was undertaken in conjunction with our
previous investigation of owner-requested surveillance for EIA
(2) and was centered on disease investigation activities within
the current Canadian EIA control program. The first objec-
tive was to describe the extent of EIA detection during disease
investigations over a 4-year period, and to examine potential
issues of importance for disease control such as the extent of
seroconversion during disease investigations and the frequency
of clinical signs in affected horses. A second objective was to
evaluate potential animal-level risk factors for EIA in this sub-
group of high-risk animals.
Materials and methods
The study covered the period from January 1, 2009 to
December 31, 2012 and the sample population comprised all
EIA cases, and all horses tested as part of an EIA disease inves-
tigation. Cases included those discovered based on sampling by
a private veterinarian accredited by the CFIA for this function
(3) and those detected during a disease investigation.
In accordance with the agency’s confidentiality rules, the
CFIA provided information on submissions for EIA testing
to the CFIA National Reference Laboratory on a sample level.
The database was limited to samples from disease investigations
and those submitted as laboratory referrals, i.e., referral for
additional testing after a non-negative competitive enzyme-
linked immunosorbent assay (cELISA) result was obtained at an
approved private laboratory. Aside from the reason for testing,
available information included the sample origin (province and
postal code), name and description of the tested horse, presence
or absence of clinical signs, dates of sample submission and
testing, applied tests [cELISA and/or agar-gel immunodiffusion
(AGID)] and their results, and anonymized, coded informa-
tion about the animal owner. For each sample, test results were
reviewed and categorized as “positive” or “negative.” If AGID
testing was performed, the AGID result was considered the final
result, and only samples positive by AGID were considered EIA
positive; results reported as “AGID atypical” were considered
negative.
Sample data were then reviewed to attribute test results to
individual horses and identify repeated testing of individuals
over time. Due to the lack of a unique national identification
system for horses, sample-level records were sorted by owner,
and available animal identifiers (breed, sex, name, age, descrip-
tion, tattoo, tag) were reviewed to identify horses tested on more
than 1 occasion, with final verification provided by the CFIA.
For horses tested more than once, seroconversion was defined
as a change in EIA test result from negative to positive during
the time frame of the study.
As the quality of information (e.g., description) varied
between repeated records for some horses, a composite record for
each horse was created for the purposes of risk factor analysis,
retaining the available information for reason for test, region of
origin, sex, breed type, coat color, age, and EIA status. Region
was recorded as “East” or “West” according to the province of
CVJ / VOL 60 / NOVEMBER 2019
N = 5271 EIA tests representing
4553 samples*
671 samples excluded
(quality control, feral horses, etc.)
N = 3882 samples representing
2746 horses
Information from 1136 repeated
samplings condensed to a single
entry per horse
N = 2746 horses
A R T I C LE
N = 2337 horses N = 409 EIA cases
EIA negative
Figure 2.  Flow diagram showing the process of moving from
sample-level information to horse-level information for the
purposes of risk factor analysis. *Some samples (n = 718) were
tested by both cELISA and AGID and only the AGID result was
retained.
origin: “East” included Ontario (ON), Quebec (QC), and the
Atlantic provinces (Nova Scotia, New Brunswick, Prince Edward
Island, Newfoundland and Labrador) and “West” included the
Yukon territory (YT), British Columbia (BC), Alberta (AB),
Saskatchewan (SK), and Manitoba (MB). Sex was recorded as
“male” or “female.” Breed type was categorized as “riding type”
(Thoroughbred, Quarter Horse, Appaloosa, Paint, Arabian,
Morgan, Fjord, and a variety of other riding horses, includ-
ing crosses in which at least 2 breeds were identified and each
individually fit into the riding type category), “draft” (Belgian,
Clydesdale, and Percheron), “pony” (Pony, Miniature, Shetland,
and Welsh), “crossbred” [horses identified as crossbreds, crosses
in which only 1 breed was identified (e.g., Quarter Horse cross),
and crosses in which the identified breeds individually belonged
to different categories (e.g., Quarter Horse-Percheron cross)],
and “other” (donkeys and mules). Coat color was categorized
as “dark” (black, brown, bay, red bay), “light” (white, gray,
palomino, cream, perlino, dunalino), and “other” (sorrel and
any other identified color). Additional color descriptors were
considered for defining the light category; qualifiers of “dark”
(e.g., dark gray) and “dappled” (e.g., dappled palomino) were
interpreted as a darkening of the basic coat color, and those
horses’ coat color was categorized as “other.” The color of any
donkey described as “gray” was also categorized as “other.” Age
categorized as 0 to 5 y, 6 to 10 y, 11 to 15 y, 16 to 20 y or
$ 21 y was based on the age at the last recorded submission. If
the birth year rather than a numerical age was provided, the age
at submission was calculated as the difference between the birth
year and the year of submission. Equine infectious anemia status
was categorized as “positive” (EIA case) or “negative” according
to the last reported test result (if horses were tested more than
once). For all variables, a defined category was assigned if incon-
gruent information for the same horse over time individually
fit into the same category (e.g., color was categorized as “dark”
when subsequent records reported color as “black” or “brown”).
Missing information, information entered as “unknown” and
incongruent information for the same horse over time that
could not be consistently assigned to 1 category was recorded as
“unknown” and excluded from the risk factor analysis.
1201
 Table 1.  Cases of equine infectious anemia (EIA) by province (2009–2012). Cases identified as
laboratory referrals were previously described (2). During the time frame of the study, EIA was not
detected in Manitoba, Ontario, and the Atlantic provinces.
Number of positives (included number of laboratory referrals)
Province/territory
or region 2009 2010 2011 2012 Cumulative
Yukon territory 23 (n = 1) 11 (n = 11) 0 14 (n = 12) 48 (n = 24)
British Columbia 14 (n = 5) 3 (n = 1) 58 (n = 1) 8 (n = 2) 83 (n = 9)
A R T I C LE

Alberta 40 (n = 4) 5 (n = 2) 5 (n = 3) 26 (n = 6) 76 (n = 15)
Saskatchewan 4 (n = 1) 0 103 (n = 10) 83 (n = 11) 190 (n = 22)
Quebec 0 2 (n = 2) 10 (n = 0) 0 12 (n = 2)
Easta 0 2 (n = 2) 10 (n = 0) 0 12 (n = 2)
Westb 81 (n = 11) 19 (n = 14) 166 (n = 14) 131 (n = 31) 397 (n = 70)
Canada 81 (n = 11) 21 (n = 16) 176 (n = 14) 131 (n = 31) 409 (n = 72)
a Ontario,Quebec, Atlantic provinces (Nova Scotia, New Brunswick, Prince Edward Island, Newfoundland and Labrador).
b Yukon territory, British Columbia, Alberta, Saskatchewan, Manitoba.

To test the association between potential risk factors (sex, age,
breed, coat color, region) and outcome, univariable statistical
analyses were completed using the STATA statistical pack-
age (STATA 13, Software; StataCorp, College Station, Texas,
USA). The association between each potential risk factor and
EIA status was examined by using mixed effects logistic regres-
sion (STATA command melogit) accounting for clustering of
horses by owner. An odds ratio (OR) with a 95% CI was used
to determine the association of each risk factor with EIA status,
and P , 0.05 was considered significant. Only variables with a
P , 0.20 were assessed for inclusion in a multivariable model,
with assessment of confounding using a 20% change in the
coefficient of interest and assessment of interaction of only
biologically relevant relationships. The relationship between age
categories and outcome was visualized by graphing the predicted
probabilities and the 95% confidence intervals. An intraclass
correlation coefficient (ICC) was calculated based on the latent
variable approach to estimate the correlation in EIA status
among horses from the same owner. Assumptions of mixed
effects models were examined by assessing residuals at the horse
and owner levels. Using commercial geographical information
system software (ArcGIS version 10.2.1; Environmental Systems
Research Institute, Redlands, California, USA), EIA cases were
mapped to the centroid of the owner’s postal code region identi-
fied by a full 6-digit postal code, or the first 5 digits of a postal
code for a small number of observations (n = 3).
Results
The database comprised information from 5271 EIA tests on
4553 samples; 718 samples were tested by both cELISA and
AGID and only the final AGID result was retained. Information
from 671 samples was excluded from the study as samples were
submitted for the purpose of quality control (n = 34), originated
from feral horses (n = 3), because the identity of the tested
horses could not be verified (n = 233) or because laboratory
referral samples ultimately tested negative and were not associ-
ated with a disease investigation (n = 401). Once repeated test-
ing of some horses over time was accounted for, the remaining
3882 samples represented the study population of 2746 animals;
409 that were categorized as EIA cases based on their last avail-
able test (404 horses, 3 donkeys, 2 mules) and 2337 that were
categorized as EIA negative (2322 horses, 12 donkeys, 3 mules,
Figure 2). Samples for 72 EIA cases (17.6%) were laboratory
referrals, i.e., they were originally taken by accredited veterinar-
ians based on owner requests, and these have been previously
described (2). The remaining 337 EIA cases were identified
during disease investigations. The overall ratio of EIA cases
discovered during disease investigations to those discovered
by owner-requested surveillance testing, therefore, was 4.7:1.
Table 1 and Figure 3 show the distribution of EIA cases across
provinces, regions, and years of the study. Most cases of EIA
were detected in the western provinces.
A total of 288 horse owners were involved in disease investi-
gations. One hundred and eighty-six owners owned only horses
that tested EIA negative, 57 owners owned a single EIA positive
horse, and 45 owners owned between 2 and 60 EIA positive
horses. The owner of the 3 affected donkeys and the owner of
1 of the mules also owned affected horses, while the second
affected mule was the owner’s only affected animal.
Based on submission dates, 49 horses seroconverted during a
disease investigation period; i.e., their EIA status changed from
negative to positive during a time period considered to represent
1 disease investigation. One additional horse seroconverted dur-
ing the time frame of the study; however, dates of testing were
approximately 20 mo apart, suggesting the horse was involved in
more than 1 disease investigation. Information on the presence
or absence of clinical signs was available for 233 EIA cases, all
of which were horses. Clinical signs were described as “present”
for 22 horses (9.4%), “possible” for 46 (19.7%), “absent” for
87 (37.3%), and “unknown” for 78 horses (33.5%). Details
concerning the nature of clinical signs were not available.
Using a mixed effects logistic regression model, which
accounted for clustering of horses by owner, the only animal-
level factor significantly associated with a positive EIA status was
age (Table 2 and Figure 4). Model fit was acceptable based on
assessment of residuals. Horses in the 3 oldest age groups had a
significantly increased risk of an EIA positive status compared
with young horses (0 to 5 y). The intraclass correlation coef-
ficient (ICC) for this model was 0.60 (95% CI: 0.49, 0.71),
and the variance at the owner level was 5.0 (95% CI: 3.2, 8.0).
Discussion
Following our previous description of the incidence of EIA detec-
tion based on owner-requested testing alone, the first objective

1202 CVJ / VOL 60 / NOVEMBER 2019

 A R T I C LE
Figure 3.  Location of cases of equine infectious anemia (EIA) identified during disease investigations from 2009 to 2012 (n = 409).
The symbols indicate the postal codes at which at least 1 EIA case was located. Most EIA positives (n = 397) were located in western
provinces (YT, BC, AB, SK) while 12 EIA positives were located in the East (QC). Also see Table 1.

of the current study was to describe the extent of EIA detection
during disease investigations and to examine issues of importance
for disease control such as seroconversion of in-contact animals
and the frequency of clinical signs in EIA positive horses at the
time of testing. In addition to the 72 EIA cases detected through
voluntary owner-requested testing at the accredited veterinary
practitioner level, disease investigations identified 337 EIA cases
(out of 2674 tested horses), indicating that most EIA cases in
Canada during the 4 years of the study were identified through
disease investigation testing. The fact that individual owners
owned from 1 to 60 EIA positive animals suggested a wide range
of degree of disease spread from individual animals, assuming
that horses belonging to 1 owner resided on the same prem-
ises. One limitation of our study is that we cannot verify this
assumption, or exclude the possibility that relationships beyond
ownership status existed between EIA cases. For instance, the
same premises may have housed horses belonging to multiple
owners, but this could not be determined from the available
data. Nonetheless, in the context of disease control, it appears
reasonable to emphasize the importance of testing potentially
exposed animals whenever a case of EIA is detected.
One limitation of the current EIA control program in Canada
is that disease investigation testing is limited to those animals “in
contact with the positive animal within 30 days of the sampling
date” (7); i.e., the extent of CFIA-requested testing is dictated
by the date disease is detected in an index case. As subclinical
carriers of the virus are common (3) and clinical signs of a first
infection may go undetected (22), the date of disease detection
does not reflect the infection date in most cases. The current
30-day disease response approach is therefore limited in its
capacity to identify all at-risk contacts, and ultimately affects
the ability to determine when a horse may have become infected
or when it may have put others at risk. Even for horses that are
tested repeatedly for EIA throughout their lifetime, estimating
the time point of infection is very difficult as, currently, there
is no mandatory unique identification of horses in Canada, and
sample and animal level data are not collected in a consistent
manner when voluntary EIA testing leads to a negative result. In
the context of our study, a time point of infection could only be
estimated for those horses that seroconverted during an ongoing
disease investigation or, in 1 case, within a 20-month period
between subsequent investigations.

CVJ / VOL 60 / NOVEMBER 2019 1203

 Table 2.  Univariable analysis of potential risk factors associated with a positive equine infectious anemia (EIA) status of horses in Canada.
Animal-level risk factors were evaluated in a subset of high-risk horses (n = 2746) using mixed effects logistic regression with a random
intercept to account for clustering of horses per owner. Missing and inconclusive data were excluded from the analysis. P-values for the
overall analysis of each factor are indicated in bold.
Factor
N = horses Number of horses Number of EIA cases P-value
(farms) Categories (% of total) (proportion positive) OR (95% CI) (Category P-value)
Sex Male 1428 (54.7) 201 (14.1) ref a 0.91
A R T I C LE

N = 2611 Female 1183 (45.3) 171 (14.5) 1.01 (0.76, 1.35)

(279)
Age 0–5 y 821 (40.8) 123 (15) ref 0.0002
N = 2011 6–10 y 763 (37.9) 127 (16.6) 1.2 (0.83, 1.73) (0.34)
(256) 11–15 y 265 (13.2) 67 (25.3) 2.6 (1.58, 4.34) (, 0.001)
16–20 y 120 (6) 28 (23.3) 2.2 (1.09, 4.48) (0.03)
$ 21y 42 (2.1) 12 (28.6) 4.4 (1.65, 11.61) (0.003)
Coat colorb Dark 879 (46.3) 131 (14.9) ref 0.58
N = 1897 Light 148 (7.8) 20 (13.5) 0.69 (0.34, 1.4)
(232) Other 870 (45.9) 143 (16.4) 0.93 (0.65, 1.32)
Breed typeb Riding 937 (59.6) 154 (16.4) ref 0.38
N = 1573 Draft 54 (3.4) 5 (9.3) 0.29 (0.09, 0.97)
(229) Pony 182 (11.6) 42 (23.1) 0.92 (0.44, 1.93)
Crossbred 380 (24.1) 40 (10.5) 0.93 (0.44, 1.99)
Other 20 (1.3) 5 (25) 1.37 (0.27, 7.03)
Regionb East 143 (5.2) 12 (8.4) ref 0.35
N = 2746 West 2603 (94.8) 397 (15.25) 2.07 (0.45, 9.55)
(288)
a ref
— reference category.
b Categories
were defined as described in the materials and methods.

Forty-nine horses (12% of all EIA cases) seroconverted dur-

Marginal predicted probabilities (mean)

ing a disease investigation period, and assessment of testing 0.4 C

dates in light of the fact that seroconversion typically occurs 0.35

within 45 d of infection (21) suggested transmission may have
occurred shortly before the disease investigation began, or may
have continued after the initial case had been detected (data not
shown). Investigation of modes of transmission was beyond the
scope of this study and, to our knowledge, modes other than
natural vector transmission were not confirmed for any of the
EIA outbreaks included in the study. When considering testing
dates, it was interesting to note that seroconversion in some
cases occurred during the late fall and winter months when
tabanid activity is expected to be low or absent. Potential expla-
nations include longer than expected seroconversion periods or
non-tabanid modes of virus transmission. Evidence of ongo-
ing seroconversion during disease investigations in this study
supports the current approach of testing until all on-premises
contact horses test negative at least 45 d after the last potential
exposure to a known EIA case.
The existence of a subclinical carrier status makes EIA a diffi-
cult disease to control without routine testing. Of 233 EIA cases
that had this information available, fewer than 10% reportedly
had clinical signs at the time of testing, although clinical signs
may have been present for an additional 19.7% of horses in
which clinical signs were described as “possible.” While report-
ing on clinical signs may be inconsistent, this suggests a large
proportion of EIA cases in this study were subclinical carriers
or, if clinical signs were present, they were not recognized at the
time of testing or not attributed to EIA. Clinical signs associated
with EIA include fever (acute or recurring), lethargy, pallor,
0.3
C BC
0.25
0.2 AB
A
0.15
0.1
0–5 6–10 11–15 16–20 211
Age category (years)
Figure 4.  Age as a significant risk factor for EIA (P = 0.0002)
based on logistic regression analysis using a mixed effects model
and accounting for clustering of horses by owner. The predicted
probabilities of EIA cases within the various age categories
are shown. Categories with different superscript letters differ
significantly from each other. Data are based on 2011 horses of
high risk, for which the age at last testing could be identified.
edema, and weight loss, with anemia and thrombocytopenia
commonly detected on hematological tests (22).
The second objective of this study was to evaluate potential
animal-level risk factors for EIA. The subset of horses used for
this analysis represented those at high risk for EIA, and this
fact has to be taken into account when interpreting our data
as the epidemiology of EIA, once the disease is introduced into
a group, may be quite different compared to its epidemiology
in the overall population. The previously reported increased
risk for an EIA positive status in the western provinces, when

1204 CVJ / VOL 60 / NOVEMBER 2019

o­ wner-requested testing was evaluated (3), disappeared once
clustering of horses by owner was taken into account in the
mixed effects model. In other words, while more outbreaks
occurred in the West during the time frame of the study, the
average number of EIA cases per owner did not differ signifi-
cantly between regions. If one assumes that horses belonging
to the same owner reside together in most cases, this finding,
as well as the high ICC, suggests that proximity to an EIA
case itself heightens the risk of EIA, which makes sense when
natural transmission of the EIA virus is considered. Conversely,
the data could also be interpreted as consistent with individual
owners’ higher-risk management practices, or lower farm-level
biosecurity measures, being associated with a higher risk for
EIA. Information about the transmission patterns in individual
disease outbreaks would be required to conclusively interpret
our data, but these were not investigated as part of this study.
The original plan was to investigate breed as a potential risk
factor reflecting management practices; however, accurate infor-
mation about breed was missing in many cases and horses of the
same breed (e.g., Thoroughbreds) may have been managed quite
differently (e.g., as riding horses or racehorses). Similar breeds,
therefore, were grouped into larger categories and based on this
grouping, breed type was not associated with disease risk. Aside
from the inherent limitations of our breed type categorization,
any effect of breed may also have been removed as individual
breeds tended to cluster by owner, again suggesting that proxim-
ity to EIA cases was a stronger risk factor than the individual
animal characteristics.
Sex was not identified as a risk factor for EIA in this study,
which is similar to previous reports (19,23,24). There are
no inherent biological reasons why disease risk should dif-
fer between the sexes; however, sex-dependent differences in
behavior or management practices might still increase the risk
of contracting EIA. In wild horse populations, an increased
risk of EIA for stallions has been suggested (25) and may be
explained by a transfer of blood or saliva during stallion fights.
Interestingly, age was a significant risk factor for EIA in our
study, with horses in the 3 oldest age groups having a signifi-
cantly increased risk compared to the youngest age category.
The data herein are consistent with 1 previous study showing
an increased risk of EIA in older horses (19) and may reflect
an increased “lifetime” risk for exposure to EIA by both natural
and iatrogenic routes in older horses. As our study evaluated
the age of EIA detection rather than exposure to the virus, our
data are also consistent with persistence of the infection for life.
An age-related reduced risk for EIA based on the observation of
reduced tabanid burdens (18) has been suggested for foals (26),
and may be explained by increased defensive behaviors. We did
not specifically evaluate disease risk in foals in this study as it
was difficult to identify foals definitively based on the informa-
tion provided in the database.
Certain host-seeking tabanids are reportedly attracted to
linearly polarized light reflected from the coat of host animals,
resulting in light-colored animals being less attractive than
darker ones (20). In addition, certain striped or spotted patterns,
which alternate from dark to bright or alternate directions of
polarization, are less attractive than homogeneous colors (21).
CVJ / VOL 60 / NOVEMBER 2019
The potential effect of light coat color was investigated in
our study, but we were not able to assess the effect of specific
color patterns as the descriptive information was not detailed
enough in most cases. Our categorization of coat colors followed
stringent criteria to try to separate out the lightest and darkest
colored horses in an attempt to mimic experimental studies;
however, coat color was not identified as a risk factor for EIA.
A R T I C LE
While this may be attributable to some of the study’s general
limitations, it is also possible that tabanid species in Canada
do not exhibit the same host-seeking behaviors as those previ-
ously studied. Tabanids may not have been the most important
contributor to disease risk during the time frame of our study,
or influences such as fly control methods were not equally dis-
tributed across groups.
This study has several limitations, which are mostly attrib-
utable to inherent difficulties when retrospectively evaluating
sample submission-based datasets. Importantly, the lack of
unique identification of horses in Canada, along with the fact
that a substantial proportion of records in the database con-
tained limited, conflicting or missing information, complicated
identification of individual horses and categorization of horses
for the purposes of risk factor analysis. Improved traceability
and identification of horses undergoing EIA testing would likely
facilitate further research, and may even be useful in the context
of disease investigations.
In conclusion, our study showed that the majority of horses
with EIA during the 4-year study period were detected based
on disease investigation testing of in-contact horses. Our data
further suggest that seroconversion of additional horses can be
expected once a disease investigation commences, and that most
horses found to have EIA in Canada are subclinical carriers.
Within a subpopulation of high-risk animals, age was identi-
fied as a significant animal-level risk factor for EIA. Additional
research is needed to investigate all potential modes of virus
transmission during disease outbreaks, and to evaluate the
impact of management practices on disease risk in Canada. CVJ
Acknowledgments
The authors gratefully acknowledge Maria Funk, Hayley
Kosolofski, and Thuy Nguyen for their assistance with data
cleaning. CVJ
References
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tive study of owner-requested testing as surveillance for equine infectious
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  3. Cook RF, Leroux C, Issel CJ. Equine infectious anemia and equine
infectious anemia virus in 2013: A review. Vet Microbiol 2013;167:
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  4. Lucas MH, Davies THR. Equine infectious anaemia. Equine Vet Educ
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in 2014: Live with it or eradicate it? Vet Clin Equine 2014;30:561–577.
A R T I C LE
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Book Review
Compte rendu de livre
Lavin’s Radiography for Veterinary
Technicians, 6th edition
Brown M, Brown LC. Elsevier, St. Louis, Missouri, USA. 2018.
627 pp. ISBN: 9780-3234-1367-1.
T his book is a great addition to any clinic’s library. It con-
tains an absolute ton of information starting with how
an X-ray machine works and where it gets its power from, to
a multitude of different specialized imaging options. It covers
the when and the how of quality control for X-ray machines,
which is great for both practices and technicians who are just
starting out. There is a fantastic group of chapters about small
animal positioning for abdomen, thorax, forelimbs, and hind
limbs, all the way to vertebrae and skull. They’ve also included
information on options for restraint and tools and tricks to
help reduce radiation exposure to staff. Sample radiographs are
provided along with footnotes to help the reader visualize and
follow along in the text.
1206
18. Foil L, Stage D, Adams WV Jr, Issel CJ. Observations of tabanid feeding
on mares and foals. Am J Vet Res 1985;46:1111–1113.
19. Borges AM, Silva LG, Nogueira MF, et al. Prevalence and risk factors for
Equine Infectious Anemia in Poconé municipality, northern Brazilian
Pantanal. Res Vet Sci 2013;95:76–81.
20. Horvath G, Blaho M, Kriska G, et al. An unexpected advantage of
whiteness in horses: The most horsefly-proof horse has a depolarizing
white coat. Proc Biol Sci 2010;277:1643–1650.
21. Egri A, Blaho M, Kriska G, et al. Polarotactic tabanids find striped pat-
terns with brightness and/or polarization modulation least attractive:
An advantage of zebra stripes. J Exp Biol 2012;215:736–745.
22. Mealey RH. Equine Infectious Anemia. In: Sellon DC, Long MT, eds.
Equine Infectious Diseases. St. Louis, Missouri: Elsevier, 2014.
23. Lucas MH, Davies THR. Equine infectious anaemia. Equine Vet Educ
1995;7:89–92.
24. Senna D, Schmitt AC, Mundim APM, Nociti DLP, Silva IS, Damasceno
MO. Epidemiological evaluation of equine infectious anaemia virus
in draught horses in Cuiaba, MT. Estudo epidemiologico do virus da
anemia infecciosa equina em cavalos de tracao, em Cuiaba — MT. Rev
Bras Med Vet 2003;25:202–204.
25. Issel CJ, Anderson G. Researchers: The stigma of EIA is misplaced.
TheHorse.com 2007. Available from: http://www.uky.edu/Ag/
VetScience/EIA/documents/178.pdf Last accessed July 31, 2019.
26. Issel CJ, Rushlow K, Foil LD, Montelaro RC. A perspective on equine
infectious anemia with an emphasis on vector transmission and genetic
analysis. Vet Microbiol 1099;17:251–286.
“Application Information” boxes and “Technician Notes” have
been added, which provide practical information for on-the-job
challenges. There was quite a bit of mention about working
with film and developing film, and while I’m sure there are still
some clinics working this way, I feel that most clinics have now
switched to digital X-ray, so in my opinion it may have been
appropriate to focus more on this aspect.
There are informative chapters on avian and exotic, and
equine and large animal radiography, which provide informa-
tion on positioning and restraint for the X-rays. The authors
also mention different tools to help with the positioning and
restraint techniques to get the best possible results. Overall I
think that this is a great book. It has educational information
and is very in-depth. I would recommend this for any veterinary
clinic because it’s always useful to be able to have a reliable and
informational resource when needed.
Reviewed by Kelly Buker, RVT, Hart Family Veterinary Clinic,
Prince George, British Columbia.
CVJ / VOL 60 / NOVEMBER 2019