Brazilian Journal of Microbiology (2020) 51:2133–2143

https://doi.org/10.1007/s42770-020-00377-z

VETERINARY MICROBIOLOGY - RESEARCH PAPER

Seroepidemiology of bovine brucellosis in Colombia’s preeminent

dairy region, and its potential public health impact
Olga Lucia Herrán Ramirez 1 & Huarrisson Azevedo Santos 1 & Ingrid Lorena Jaramillo Delgado 2 &
Isabele da Costa Angelo 1

Received: 25 June 2020 / Accepted: 4 September 2020 / Published online: 12 September 2020
# Sociedade Brasileira de Microbiologia 2020

Abstract
A cross-sectional study was conducted to determine the associated factors of brucellosis in Colombia’s preeminent dairy region
declared in quarantine. A total of 656 samples were collected from cows ≥ 2-year-old from 40 herds. Samples were screened by
the Rose Bengal Plate Test, and the Fluorescence Polarized Assay test and Competitive ELISA were used as confirmatory tests.
A cow was classified as positive if the screening and both confirmatory tests were positive. A herd was classified as positive if at
least one cow was seropositive. The factors associated to seropositivity were tested using a logistic regression model with
explanatory variables regarding cattle management, zootechnical parameters, and sanitary practices. The seroprevalence at the
animal level was 6.6% (43/656) and at herd level 27.5% (11/40). In the model, five variables explained the animal cases: purchase
or animal transfer between owner’s farms (OR = 2.79, 95% CI 1.42, 5.49), history of abortion (OR = 4.22, 95% CI 1.91, 9.33),
birth of weak calves (OR = 13.77, 95% CI 2.75, 68.91), use of a bull for mating (OR = 9.69, 95% CI 2.23, 42.18), and the
vaccination in adulthood (OR = 3.03, 95% CI 1.04.8.78). In the model at the herd level, two variables explained the cases: birth of
weak calves (OR = 9.60, 95% CI 1.54, 59.76) and purchase or animal transfer between owner’s farms (OR = 7.22, 95% CI 1.03,
50.62). These results justify the need for a quarantine declaration in the region and the implementation of epidemiological studies
as a public health measures used to combat outbreak.

Keywords Brucellosis . Bovine . Dairy . Outbreak . Public health . Seroepidemiology

Introduction animals, accidental inoculation during vaccination programs,

Brucellosis is a neglected zoonotic disease caused by the
Brucella spp. The infection causes a severe, chronic, and de-
bilitating disease in humans. People can become accidental
hosts of Brucella spp. by the consumption of animal-source
foods without adequate cooking and pasteurization processes
[35]. Brucellosis is an occupational disease that can be trans-
mitted by direct contact with the secretions of infected
Responsible Editor: Roxane Piazza
* Olga Lucia Herrán Ramirez
luciaherranmvz@gmail.com
1
Department of Epidemiology and Public Health, Veterinary Institute,
Federal Rural University of Rio de Janeiro, P.O. Box 23897-000,
Seropedica, RJ, Brazil
2
Faculty of Graduate School, CES University, P.O. Box 050021,
Medellin, Colombia
or through airborne transmission to the vaccinator, veterinar-
ians, laboratorians, slaughterhouse workers, and field
workers. Therefore, eradicating the disease has become a ma-
jor goal of public health programs in endemic countries [49].
Bovine brucellosis is found across the world; it causes pro-
duction losses with significant financial impact to farmers
[44]. The main risk factors for the introduction and spread of
brucellosis in livestock are directly related to the lack of
biosecurity and best management practices [42]. Brucellosis
is usually asymptomatic in young animals and non-pregnant
females. In pregnant females, the principal symptom is abor-
tion [50]. However, even in the absence of an abortion, the
bacteria can easily spread to cattle herds through a profuse
excretion of organisms in the placenta, fetal fluids, and vaginal
discharges [28]. The bacteria can live in soil, water, pasture,
and manure for a long time [26]. The excretion of Brucella in
the environment is a risk to the public health of rural families
[44].
2134
The “gold standard” for a definitive diagnosis is a culture of
Brucella spp., but the diagnostic sensitivity is low, and the
bacteria is considered a bioterrorist agent [36, 45].
Therefore, the OIE recommends serological testing for the
surveillance and the development of public health programs
in developing countries [36]. A few studies with Brucella
isolation have been conducted in Colombia: B. abortus
(biovars 1, 2, and 4) [30, 40], B. canis [38], and B. suis (biovar
1) [41, 47]. However, official data only recognizes B. abortus
[20].
There are currently no vaccines that can prevent brucellosis
in humans [9]. The smooth strain S19 and the rough strain
RB51 vaccines are used in livestock as a tool for epidemio-
logical control. However, both vaccines have drawbacks: The
RB51 strain is virulent for humans and resistant to rifampicin,
an antibiotic used for treating human brucellosis. In addition,
vaccinated cattle can, in some cases, shed the vaccine-strain
into their milk [33]. Finally, some antibodies produced by
vaccinated cattle can be detected in serological testing and
give a false positive result [36].
In the last couple of years, Colombia’s preeminent
dairy region has seen an unusual increase in their num-
ber of seropositive cattle. This made it necessary to
declare a quarantine for brucellosis and implement of
sanitary measures for brucellosis control in the region
[22, 24]. Our study addresses the prevalence of bovine
brucellosis from Colombia’s dairy region and its factors
associated with the increase of seropositivity as a seri-
ous public health problem.
Materials and methods
Study design
Between January and April 2019, a cross-sectional study was
conducted to determine the prevalence of Brucella spp. anti-
bodies and the factors associated with seropositivity in dairy
herds from the Antioquia region in Colombia.
Study area
The study was conducted in four municipalities of Antioquia:
Santa Rosa de Osos (6° 38′ 50″ N, 75° 27′ 38″ O),
Entrerrios (6° 33′ 55″ N, 75° 30′ 51″ O), Belmira (6°
36′ 18″ N, 75° 39′ 57″ O), and San Pedro de los
Milagros (6° 27′ 34″ N, 75° 33′ 28″ O) (Fig. 1). This
region is the leading milk production in Colombia. It
represents about 30% of the country’s annual milk pro-
duction. Around 31,250 families make their living from
the production of milk in this region [11].
Braz J Microbiol (2020) 51:2133–2143
Study population, sampling, and epidemiological
data collection
The target population was dairy cattle of Antioquia, within the
selected municipalities. The number of cows was defined ac-
cording to the data of the vaccination campaign for the Foot
and Mouth Disease and Brucellosis [23]. Dairy cows ≥ 2-year-
old in lactation or dry period were the primary sampling units.
Dairy herds in compliance with brucellosis vaccine programs
were considered secondary sampling units. The formula n =
1.962 p (1 − p)/d2 was used to calculate the sample size with a
confidence level of 95%, desired absolute precision (d2) of
5%, and expected prevalence (p) of 10% [46]. Proportionate
sampling was used from each municipality to ensure that the
municipalities with large numbers of dairy cattle were not
being under-represented. Farms were selected by simple ran-
dom sampling using a random number table without replace-
ment. Herd managers were asked to participate in the study,
and they voluntarily signed the informed consent agreement to
participate. The concept of aggregated sensitivity (Se) and
specificity (Sp) was used [16, 43] in order to allow the sample
size in the herd to detect at least one positive animal with 95%
confidence [43]. A total of 15 dairy cows were sampled in
herds with ≤ 50 cows, whereas 22 cows were sampled in larg-
er herds (> 50 cows). The random selection was made from
the available list of animals. For the blood collection, the skin
area was cleaned with chlorhexidine, and using blood collec-
tion tubes with anticoagulant, six milliliters of venous blood
were taken from each animal. In order to allow clot separation,
all blood tubes were let to stand at ambient temperature, then
refrigerated and transported to the laboratory properly.
Before the sampling collection, a survey was given to the
farmers. The survey was based on a review of literature and
included closed questions related to the origin of the cattle
(born at the farm, purchased or animal transfer between
owner’s farms), the disposal of waste birth, vaccination pro-
tocols (calf 3–8 months, heifers, and cows), co-grazing with
another species, sharing of water sources, presence of black
vultures and their access to birth waste, methods of disposal
for discarded milk (pen, septic tank, calf), type of vaccine
strain used (strain RB51, strain S19, both strain), brucellosis
status of the herds (free, unknown, positive), milking process
(pen, mechanic system in pen, parlor milking), mating type
(artificial insemination (AI), bull, both), and other variables
that may be associated with bovine brucellosis. Additionally,
individual questionnaires were conducted to identify variables
such as origin of the cow, age (24–36 month, > 36–60 month,
> 60 month), month of gestation (neither, 1–90 days, >
90 days), number of births (0–1 calf, 2–4 calves, > 4 calves),
vaccination boosters, reproductive problems (abortion, birth
of weak calves, retention of fetal membranes). and to confirm
the mating type. Both the survey and questionnaire were pre-
Braz J Microbiol (2020) 51:2133–2143
Fig. 1 Map of Antioquia,
Colombia, showing the 4
municipalities from the north of
Antioquia where this study was
conducted
tested and validated by farm managers that did not belong to
the study.
Laboratory diagnostics
The Rose Bengal Plate Test (RBT) was used as the screening
test and processed at a laboratory authorized by the
Colombian Agricultural Institute (ICA) for diagnosis of
brucellosis (Approval number 027-2016). The agglutina-
tion denoted a positive test for RBT and vice versa. The
samples that did not have an agglutination were consid-
ered negative. Positive RBT samples were confirmed by
the Fluorescence Polarized Assay test (FPA) and
Competitive ELISA (cELISA) in the Laboratory ICA.
The FPA results were expressed in millipolarizations
(mP). A sample was considered positive if it was equal
or greater than 121 mP. Samples equal or lower than
107 mP were considered negative, and those between
108 to 120 mP were considered suspects. The cElisa
results were obtained by comparing the sample optical
density with the positive control mean optical density
(OD) to 450 nm. For the interpretation of the results,
samples with a percentage inhibition (PI) ≥ 30% were
considered positive for the presence of Brucella anti-
bodies and those with PI < 30% were considered nega-
tive. All serological tests were performed and
interpreted according to standardized procedures OIE
[36]. Combinations of serological tests were used to
improve Se and Sp, complying with the official regula-
tions of the Control, Prevention and Eradication
Program of Brucellosis in Colombia [21].
2135
Case definition
At the animal level, the case definition was a dairy cow ≥ 2-
year-old with positive results in the RBT diagnosis test, and
then positive to both confirmatory tests. A herd was classified
as positive if at least one cow was seropositive.
Statistical analysis
With subsequent serial testing, the seroprevalence of Brucella
spp. and confidence intervals (CI) were determined with a Se
(98%) and Sp (99.7%) according to ICA [21]. A chi-square
test was used to evaluate the association between Brucella
spp. seropositivity at the animal and the herd level (binary
outcome: seropositive/seronegative) and all explanatory vari-
ables. A p < 0.05 was considered to be statistically significant.
Variables statistically significant and variables with a p < 0.25,
but important according to other epidemiological studies,
were offered to the logistic regression models at the animal
and the herd level. The variables included in the models were
based on forward method “wald,” which tests the significance
of inclusion for the variable at each stage and using at 0.1 as
cut point. Prior to building a final model, the collinearity be-
tween independents variables was tested through cross tabu-
lation using Fisher’s exact test. When two variables were
found with high collinearity, only the variable that had more
relevance according to literature review was included in the
model. The validity of the model to the observed data was
assessed by the Hosmer–Lemeshow goodness-of-fit test.
(p > 0.05). Finally, the coefficients were estimated, and the
appropriateness adequacy and usefulness of the model were
2136
identified. The statistical analysis of the data was performed
with the statistical program IBM SPSS (Statistical Package for
the Social Sciences) v. 25.0 for Windows (Armonk, Nueva
York: IBM Corp.).
Ethics approval
The study was approved by the Research Ethics Committee
(CEP Approval number 1.243/18) and by the Ethics
Committee for Animal Use (CEUA Approval number
8427171219) for the Veterinary Institute of Rural Federal
University of Rio de Janeiro (UFRRJ).
Results
The sera of 656 bovines were collected from 40 dairy herds;
with subsequent serial testing, the overall animal-level sero-
prevalence was 6.6% (43/656; 95% CI 4.8, 8.7). The herd-
level seroprevalence was 27.5% (11/40; 95% CI 14.6, 43.9)
Within the herd, the mean of positive animals was 3.91 (range
1.0, 10.0).
In the univariable analysis, all significant variables at 5%
(Tables 1 and 2) were included to the logistic regression anal-
ysis. The variable retention of fetal membranes at the animal-
level, as well as brucellosis status of the herds, history of
abortion, and origin of the cattle at the herd level were not
significant (p < 0.25). However, they were considered impor-
tant and included in the logistic regression analysis.
In the logistic regression model at the animal-level, five
variables correctly explained 73.8% of the cases: origin of
the cow (purchased or animal transfer between owner’s farms)
(OR = 2.79, 95% CI 1.42, 5.49), history of abortion (OR =
4.22, 95% CI 1.91, 9.33), the birth of weak calves (OR =
13.77, 95% CI 2.75, 68.91), the use of a bull for mating
(OR = 9.69, 95% CI 2.23, 42.18), and use of the RB51 vac-
cine strain in the animal adulthood (OR = 3.03, 95% CI 1.04,
8.78). All variables showed a p < 0.05, which affirms that the
independent variables influence the dependent variable (sero-
positive animal). The Hosmer–Lemeshow goodness-of-fit test
showed that the model fit the data (X2 = 2.73, d.f. 6, p = 0.841)
(Table 3). In the logistic regression model at the herd level,
two variables correctly explained 82.5% of the cases: the birth
of weak calves (OR = 9.60, 95% CI 1.54, 59.76) and the ori-
gin of the cattle (purchased or animal transfer between
owner’s farms) (OR = 7.22, 95% CI 1.03, 50.62). All vari-
ables showed a p < 0.05, which affirms that the independent
variables influence the dependent variable (seropositive herd).
The Hosmer–Lemeshow goodness-of-fit test showed that the
model fit the data (X2 = 0.40, d.f. 2, p = 0.81) (Table 3).
The Cramer phi prime estimates indicated no important
correlations between any pairs of the independent variables.
Braz J Microbiol (2020) 51:2133–2143
None of the pair-wise interactions were statistically signifi-
cant, and there was no evidence of confounding bias.
Discussion
The purpose of this study was to determine the seroprevalence
of bovine brucellosis and their factors associated in a dairy
region of Colombia. The study showed an increase compared
with the average prevalence reported by the ICA between
2005 and 2016, both at the department (4.3% at the animal
level and 25.9% at the herd level) and at the national level
(4.4% at the animal level and 22% at the herd level) [5, 20].
The prevalence results were in line with the quarantine man-
date in the region of study [22, 24]. Nevertheless, the difficulty
to be able to identify by the use of serology, the type Brucella
spp. antibodies induced (wild-type or vaccinal strains), is a
great limitation for the development of effective strategies of
control and eradication of brucellosis, and the proper analysis
of the outbreaks [19].
Bovine brucellosis spreads easily in dairy regions because
there are favorable environmental conditions for their prolif-
eration. The different management practices used for milk
production offer an opportunity for the bacteria to multiply
and spread the disease faster [10]. Consistent with others stud-
ies, management practices such as co-grazing [15, 17], the
sharing of water sources with the neighborhood [8] and the
lack of exclusive maternity pens [15] were associated with a
seropositive animal in the study.
Parlor milking was associated to seropositivity at the ani-
mal-level. The authors who reported similar results attributed
this association to the close contact between the animals,
which increases the longer the cows are part of the milking
process [3, 7, 17]. The use of discarded milk to feed calves
showed significant association with seropositivity at the ani-
mal level. Potential secretion of viable bacteria through mam-
mary glands was shown in a study that isolated B. suis from
milk of cows naturally infected during a period of 2 years [18].
Similarly, other studies have shown the potential shedding of
RB51 and S19 strain into the cows’ milk [12, 35]. In 2018, the
Center for Disease Control and Prevention (CDC) studied
three cases of brucellosis in humans. The individuals had
contracted brucellosis by the intake of unpasteurized milk
and were isolated RB51 strain [12, 35].
The seropositivity at the animal level also showed a signif-
icant association with the presence of wildlife, which is con-
sistent with other studies conducted in Brazil, Zimbabwe, and
the USA [15, 34, 42]. Surveillance of wildlife and
synanthropic animals is strongly recommended, since those
animals could serve as potential reservoirs or vectors [19,
29]. The presence of black vultures and opossums was com-
mon at most of the farms. However, there was a significant
association between the presence of rodents and a seropositive
Braz J Microbiol (2020) 51:2133–2143 2137

Table 1 Univariable analysis at the animal level between independent variables and Brucella spp. seropositive of 656 cows in 40 herds located in dairy
cattle of Antioquia, Colombia

Animal level = 656

Variable Frequency % p** OR (IC 95%)

Origin 0.010 2.261 (1.19, 4.28)

Purchased or transferred 27/289 62.8
Born 16/367 37.2
Age 0.805
24–36 months 10/150 23.3
37–60 months 15/258 34.9
> 60 month 18/248 41.9
Month of gestation 0.836
Neither 17/257 39.5
1–90 days 11/147 25.6
> 90 days 15/252 34.9
Number of pregnancies 0.764
Neither 2/20 4.7
1–2 18/279 41.9
3–4 12/217 27.9
>4 11/140 25.6
Abortion 0.001 3.78 (1.80, 7.96)
Yes 11/62 25.6
No 32/594 74.4
Retention of fetal membranes 0.183 2.941 (0.62, 13.87)
Yes 2/12 4.7
No 41/644 95.3
Birth of weak calves 0.023 6493 (1.61, 26.06)
Yes 3/10 7.0
No 40/646 93.0
Disposal of waste birth 0.000 0.123 (0.05, 0.28)
Neither 33/624 76.7
Buried 10/32 23.3
Maternity pen 0.001 3.354 (1.53, 7.34)
No 35/382 81.4
Yes 8/274 18.6
Co-grazing 0.020 3.730 (1.13, 12.24)
Yes 40/519 93.0
No 3/137 7.0
Shared water source 0.000 4.984 (2.07, 11.98)
Yes 37/376 86.0
No 6/280 14.0
Mating type 0.000
AI 2/186 4.7
Bull 11/263 25.6
Both 30/207 69.8
Booster vaccination in adulthood 0,026 2.004 (1.07, 3.73)
Yes 24/261 55.8
No 19/395 44.2
Disposal of discarded milk 0.050
Calf 27/296 62.8
2138 Braz J Microbiol (2020) 51:2133–2143

Table 1 (continued)

Animal level = 656

Variable Frequency % p** OR (IC 95%)

Septic tank 4/105 9.3

Pen 12/255 27.9
Health status 0.000
Positive status 25/157 58.1
Brucellosis free status 6/264 14.0
Unknown Status 12/235 27.9
Parlor Milking 0.007 2.509 (1.26, 4.97)
Yes 31/342 72.1
No 12/314 27.9
Milking type 0.009
Parlor Milking 31/342 72.1
Mechanic milking system in pen 12/247 27.9
Milking in pen 0/67 0.0
Access of the black vultures to birth or abortion 0.001 0.255 (0.11, 0.55)
Yes 33/602 76.7
No 10/54 23.3
Black vulture, opossums, and rodents 0.000 3.621 (1.70, 7.67)
Yes 34/347 79.1
No 9/309 20.9
Wildlife 0.000
Black vultures (BV) 8/134 18.6
BV and opossums 0/74 0.0
BV, opossums, rodents 34/347 79.1
BV, opossums, other 1/90 2.3
Neither 0/11 0.0

% percent of positive, p value statistical significance, OR odds ratio, (95% CI) confidence interval with p** < 0.05

animal. This is supported by a study in central Argentina that
evidenced Brucella spp. antibodies in rodents and opossums
captured in dairy farms [29]. The presence of Brucella infec-
tion in rodents should not be underestimated and should be
explored in detailed as a risk factor in rural environments.
In this study, the access of the black vultures to birth or
abortion wastes appear to be associated protective factors in
this study. Black vultures can prevent the spread of bacteria in
the environment because they ingest afterbirth or abortion
wastes within 24 h of the event [14, 31, 42]. It can be faster
than finding and disposing the waste properly by the farmer.
At this time, it has not been determined that black vultures
increase the risk of transmission of brucellosis in livestock,
rather it appears to reduce the risk and likely act as a biological
control agent of disease [4, 31, 48].
The majority of the factors associated with the introduction
and spread of brucellosis in livestock are directly related to the
management practices [42], being the introduction of new
animals to the herds, either purchased or transferred, the most
predominantly factor. This has been proven by this study and
widely reported by other epidemiological studies [8, 15, 32].
The presence of abortion and birth of weak calves contrib-
uted to explain the seropositivity and should generate alerts,
being the birth of weak calves the more likely related to sero-
positivity. Most studies highlight the presence of abortion as a
warning sign for brucellosis [1, 6, 50]. However, even in full-
term pregnancies, the bacteria can easily spread to environ-
ment through placenta excretion, fetal fluids, and vaginal dis-
charges [28]. Appropriate control mechanisms as maternity
pens and biosecurity management best practices should also
be present in farms to prevent the spread of the infection.
Bulls can be infected by the bacteria, by errors during vac-
cination of the calves, or by exposure to a contaminated envi-
ronment [25, 39]. The B. abortus specie can be present in the
testicles and the genital glands of the bulls [13]. Transmission
can occur by infected semen, either using a bull or through
artificial insemination [2, 25]. The use of a bull, AI or both,
were significantly associated with the seropositivity at the
Braz J Microbiol (2020) 51:2133–2143 2139

Table 2 Univariable analysis at the herd level between independents variables and Brucella spp. seropositive of 656 cows in 40 herds located in dairy
cattle of Antioquia, Colombia

Herd level = 40

Variable Frequency % p** OR (IC 95%)

Origin 0.073 5.538 (1.01, 30.25)

Purchased or transferred 9/22 81.8
Born 2/18 18.2
Abortion 0.159 0.667 (0.52, 0.84)
Yes 11/33 100
No 0/7 0.0
Retention of fetal membranes 0.486 0.677 (0.41, 8.59)
Yes 8/25 72.7
No 3/15 27.3
Birth of weak calves 0.014 7.500 (1.53, 36.71)
Yes 6/10 54.5
No 5/30 45.5
Disposal of waste birth 0.479 0.357 (0.02, 6.26)
Neither 10/38 90.9
Buried 1/2 9.1
Maternity pen 0.486 1.882 (0.41, 8.59)
No 8/25 72.7
Yes 3/15 27.3
Co-grazing 0.405 3.182 (0.34, 29.43)
Yes 10/32 90.9
No 1/8 9.1
Shared water source 0.473 2.167 (0.47, 9.86)
Yes 8/24 72.7
No 3/16 27.3
Mating type 0.419
AI 2/10 18.2
Bull 4/18 36.4
Both 5/12 45.5
Type of vaccine strain 0.623
Strain RB51 9/29 81.8
Strain S19 1 /4 9.1
Both strain 0/4 0.0
Neither 1/3 9.1
Disposal of discarded milk 0.332
Calf 7/18 63.6
Septic tank 1/7 9.1
Pen 3/15 27.3
Health status 0.078
Positive status 5/9 45.5
Brucellosis free status 4/16 36.4
Unknown Status 2/15 18.2
Parlor Milking 0.728 1.354 (0.33, 5.55)
Yes 5/16 45.5
No 6/24 54.5
Milking type 0.338
Parlor Milking 6/19 54.5
2140 Braz J Microbiol (2020) 51:2133–2143

Table 2 (continued)

Herd level = 40

Variable Frequency % p** OR (IC 95%)

Mechanic milking system in pen 5/16 45.5

Milking in pen 0/5 0.0
Access of the black vultures to birth or abortion 1.000 0.741 (0.06, 9.09)
Yes 10/37 90.9
No 1/3 9.1
Black vulture, opossums, and rodents 1.000 0.921 (0.22, 3.92)
Yes 7/26 63.6
No 4/14 36.4
Wildlife 0.697
Black vultures (BV) 3/8 27.3
BV and opossums 4/18 36.4
BV, opossums, rodents 3/7 27.3
BV, opossums, other 1/6 9.1
Neither 0/1 0.0

% percent of positive, p value statistical significance, OR odds ratio, (95% CI) confidence interval with p** < 0.05

animal-level. In the logistic regression model, the cows mated
with a bull explained a seropositive animal. The results of this
study were similar to a study in Bangladesh [1] and to a recent
case control study in Colombia where natural breeding with
bulls with an unknown health status showed a higher risk,
compared with the farms that used AI [8].
Calfhood vaccination is part of regular health control pro-
grams in endemic countries [36]. In Colombia, the S19 strain
only requires a single vaccination between the 3 and 8 months
of age. When the RB51 strain is used, animals are required to
be revaccinated in adulthood (heifer and cows) [21]. A milk
discard period has not been established for this vaccine in
dairy cattle and can represent a risk factor for farmers and
lactating calves. From the cattle evaluated, the RB51 strain
was the vaccine more predominantly used. The RB51 vaccine
applied in adulthood has a significant association with
Brucella spp. seropositive at the animal level. The quarantine
status of the region may highly influence a booster vaccina-
tion. These results should be interpreted with caution, espe-
cially in endemic countries where the prevalence changes [37]
and may increase the animal susceptibility and decrease the
efficacy and the protective effect of the vaccine [27]. Another
potential interpretation is that strain antibodies in both vac-
cines S19 and RB51 can persist into adulthood and a positive

Table 3 Multivariable models with potential risk factors at the animal and the herd level for Brucella spp. seropositivity in dairy cattle of Antioquia,
Colombia

Animal level = 656 Herd level = 40

Factors Category Β S.E (β) p** OR (IC 95%) β S.E (β) p** OR (IC 95%)

Mating type Bulls 2.27 0.775 0.002 9.69 (2.23, 42.18)

AI ref.
Origin Purchased or transferred 1.02 0.345 0.003 2.79 (1.42, 5.49) 1.977 0.993 0.047 7.223 (1.03, 50.62)
Born ref.
Abortion Yes 1.44 0.40 0.000 4.22 (1.91, 9.33)
No ref.
Weak calves Yes 2.62 0.82 0.001 13.77 (2.75, 68.91) 2.262 0.933 0.015 9.606 (1.54, 59.76)
No ref.
Booster vaccination Yes 1.09 0.543 0.043 2.99 (1.03, 8.58)
No ref.
Constant -6.42 0,938 0,000 −2.97 0.971 0.002

Dependent variable: positive animal, results given with β: beta; S.E: standard errors; OR: odds ratio; (95% CI): confidence interval with p** < 0.05
Braz J Microbiol (2020) 51:2133–2143
serological cross-reaction could occur [36]. Therefore, the loss
of false-positive animals should be taken into consideration.
Conclusion
This study confirms an increase in the brucellosis seropreva-
lence justifying the associated quarantine declaration in the
Colombia’s preeminent dairy region. Factors such as the in-
troduction of new animals, the presence of abortion, the birth
of weak calves, use of bulls as a reproductive practice, and the
use of RB51 vaccine applied in adulthood were all high risk
factors associated with seropositivity. Farmers should be par-
ticularly sensitive to these factors and use them with in con-
junction with serological testing to confirm the presence of
infected cattle.
Brucellosis can have a severe impact in human’s health. It
is vital to develop effective strategies to prevent and eradicate
the infection in livestock. Adequate vaccination programs, the
implementation of preventive measures to manage risk fac-
tors, permanent surveillance with epidemiological studies,
and the correct diagnosis of suspected cases will improve food
safety and public health.
Acknowledgments We would like to thank the Colombian Agricultural
Institute and the Federal Rural University of Rio de Janeiro for facilitating
the execution of this study. We also thank TestLab S.A.S for their col-
laboration with the laboratory process. Finally, special thanks to
Corpogansa del Norte of Antioquia for their support with our fieldwork,
especially their associates for allowing us to conduct the research on their
cattle.
Authors’ contributions OLH: conceptualization, methodology, formal
analysis, investigation, writing, review and editing. HAS and IDA: con-
ceptualization, methodology, review, editing, supervision and project ad-
ministration and funding acquisition. ILJ: participated in the methodolo-
gy, review and editing.
Funding This work was supported by the Coordination for the
Improvement of Higher Education Personnel (CAPES, finance code
001). This work was financial supported, by the National Council for
Scientific and Technological Development (CNPq) for the fellowship
granted to HAS (Research Productivity Scholarship, grant number
310819/2018-0) and the Foundation Carlos Chagas Filho Research
Support of the State of Rio de Janeiro (FAPERJ).
Data availability Not applicable
Compliance with ethical standards
Conflict of interest The authors declare that they have no conflict of
interest.
Ethical approval The study was approved by the Research Ethics
Committee (CEP Approval number 1.243/18) and by the Ethics
Committee for Animal Use (CEUA Approval number 8427171219) for
the Veterinary Institute of Rural Federal University of Rio de Janeiro
(UFRRJ).
2141
Consent to participate Herd managers voluntarily signed the informed
consent agreement to participate.
Consent for publication Not applicable
Code availability Not applicable
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