EPIDEMIOLOGIC AND ANTIBIOTIC RESISTANCE PROFILE AND DETECTION OF MECA GENE IN STAPHYLOCOCCUS SPP. ISOLATES FROM DOG-OWNING VETERINARY STUDENTS

Abstract

The maintenance of pets as reservoirs of multiresistant bacteria and the transmission of microorganisms such as Staphylococcus spp. between animals and humans can affect the effectiveness of antimicrobials in human medicine. The aim of this study was to detect risk factors, evaluate the phenotypic profile of antimicrobial resistance and detect the mecA gene in Staphylococcus spp. isolated from the nasal cavity of students of veterinary medicine who own dogs. This is a field survey where 35 nasal swab samples were collected to isolate Staphylococcus spp. The antimicrobial resistance of the isolates and the classification according to the multidrug resistance profile (MDR) were determined. The presence of the mecA gene was investigated in isolates with resistance to oxacillin. In addition, the research subjects answered a questionnaire about behavior towards the dog and hygiene habits to identify risk variables for developing antimicrobial resistance. The antimicrobials tested were ampicillin, penicillin, oxacillin, cephalothin, clindamycin, gentamicin, erythromycin, enrofloxacin, and tetracycline. 92.9% of coagulase-positive staphylococci (CoPS) and 45% of coagulase-negative staphylococci (CoNS) were resistant to the beta-lactam class, and 28.6% of CoPS and 45% of CoNS showed MDR profile. Three isolates were classified as resistant to oxacillin, and the mecA gene was detected in 100% of these isolates. About half of the individuals used antimicrobials in the last 12 months (52.9%), and 75% used amoxicillin, which could explain the high antimicrobial resistance profile. Dog owners harbor Staphylococcus spp. with high resistance to beta-lactam antimicrobials and a multi-resistance profile, representing a unique One Health problem.

Full text

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EPIDEMIOLOGIC AND ANTIBIOTIC RESISTANCE PROFILE
AND DETECTION OF MECA GENE IN STAPHYLOCOCCUS SPP.
ISOLATES FROM DOG-OWNING VETERINARY STUDENTS
Recebido em: 21/08/2023
Aceito em: 18/09/2023
DOI: 10.25110/arqsaude.v27i9.2023-006
Paula Santina Banhe Cabral
1
Daniela Dib Gonçalves
2
Juliana Silveira do Valle
3
Maria Augusta Dorigan Bondezan
4
Rafaela Galves Ferreira
5
Izabela Camilotti Dorneles
6
Izabela Carvalho dos Santos
7
Luciana Kazue Otutumi
8
ABSTRACT: The maintenance of pets as reservoirs of multiresistant bacteria and the
transmission of microorganisms such as Staphylococcus spp. between animals and
humans can affect the effectiveness of antimicrobials in human medicine. The aim of this
study was to detect risk factors, evaluate the phenotypic profile of antimicrobial resistance
and detect the mecA gene in Staphylococcus spp. isolated from the nasal cavity of students
of veterinary medicine who own dogs. This is a field survey where 35 nasal swab samples
were collected to isolate Staphylococcus spp. The antimicrobial resistance of the isolates
and the classification according to the multidrug resistance profile (MDR) were
determined. The presence of the mecA gene was investigated in isolates with resistance
to oxacillin. In addition, the research subjects answered a questionnaire about behavior
towards the dog and hygiene habits to identify risk variables for developing antimicrobial
resistance. The antimicrobials tested were ampicillin, penicillin, oxacillin, cephalothin,
clindamycin, gentamicin, erythromycin, enrofloxacin, and tetracycline. 92.9% of
coagulase-positive staphylococci (CoPS) and 45% of coagulase-negative staphylococci
(CoNS) were resistant to the beta-lactam class, and 28.6% of CoPS and 45% of CoNS
showed MDR profile. Three isolates were classified as resistant to oxacillin, and the mecA
gene was detected in 100% of these isolates. About half of the individuals used
antimicrobials in the last 12 months (52.9%), and 75% used amoxicillin, which could
1
Master in Animal Science. Postgraduate in Animal Science with Emphasis on Bioactive Products.
Universidade Paranaense (UNIPAR). E-mail: paula.cabral@edu.unipar.br
2
Postgraduate in Animal Science with Emphasis on Bioactive Products and Postgraduate in Medicinal
Plants and Phytotherapics in Primary Care. Universidade Paranaense (UNIPAR).
E-mail: danieladib@prof.unipar.br
3
Postgraduate in Animal Science with Emphasis on Bioactive Products and Postgraduate in Biotecnology
Applied to Agriculture. Universidade Paranaense (UNIPAR). E-mail: jsvalle@prof.unipar.br
4
Student of Animal Science with Emphasis on Bioactive Products. Universidade Paranaense (UNIPAR).
E-mail: mariadorigan.vet@gmail.com
5
Student of Animal Science with Emphasis on Bioactive Products. Universidade Paranaense (UNIPAR).
E-mail: rafaela.ferreira@edu.unipar.br
6
Student of Animal Science with Emphasis on Bioactive Products. Universidade Paranaense (UNIPAR).
E-mail: dorneles.izabela@gmail.com
7
Doctor in Animal Science with Emphasis on Bioactive Products. Universidade Paranaense (UNIPAR).
E-mail: isacarsan@gmail.com
8
Postgraduate in Animal Science with Emphasis on Bioactive Products. Universidade Paranaense
(UNIPAR). E-mail: otutumi@prof.unipar.br

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explain the high antimicrobial resistance profile. Dog owners harbor Staphylococcus spp.
with high resistance to beta-lactam antimicrobials and a multi-resistance profile,
representing a unique One Health problem.
KEYWORDS: Pets; Epidemiology; Resistance Genes; One Health.
PERFIL EPIDEMIOLÓGICO E DE RESISTÊNCIA A ANTIBIÓTICOS E
DETECÇÃO DO GENE MECA EM STAPHYLOCOCCUS SPP. ISOLADOS DE
ESTUDANTES VETERINÁRIOS PROPRIETÁRIOS DE CÃES
RESUMO: A manutenção de pets como reservatórios de bactérias multirresistentes e a
transmissão de microrganismos como Staphylococcus spp. entre animais e humanos
podem afetar a eficácia de antimicrobianos na medicina humana. O objetivo deste estudo
foi detectar fatores de risco, avaliar o perfil fenotípico de resistência antimicrobiana e
detectar o gene mecA em Staphylococcus spp. isolados da cavidade nasal de estudantes
de medicina veterinária proprietários de cães. Trata-se de pesquisa de campo onde 35
amostras de swab nasal foram coletadas para isolar Staphylococcus spp. A resistência
antimicrobiana e a classificação segundo o perfil de multirresistência (MDR) dos isolados
foram determinadas. A presença do gene mecA foi investigada em isolados resistentes à
oxacilina. Os estudantes responderam a um questionário sobre comportamento em
relação ao cão e hábitos de higiene para identificar variáveis de risco para o
desenvolvimento de resistência antimicrobiana. Foram testados os antimicrobianos
ampicilina, penicilina, oxacilina, cefalotina, clindamicina, gentamicina, eritromicina,
enrofloxacina e tetraciclina. 92,9% dos estafilococos coagulase-positivos (CoPS) e 45%
dos estafilococos coagulase-negativos (CoNS) foram resistentes à classe dos beta-
lactâmicos e 28,6% CoPS e 45% CoNS apresentaram perfil MDR. Três isolados foram
classificados como resistentes à oxacilina e o gene mecA foi detectado em 100% destes
isolados. Mais da metade dos indivíduos fez uso de antimicrobiano nos últimos 12 meses
(52,9%), 75% fizeram uso de amoxicilina, o que poderia explicar o alto perfil de
resistência antimicrobiana. Os donos de cães abrigam Staphylococcus spp. com alta
resistência aos antimicrobianos beta-lactâmicos e apresentam MDR, o que representa um
problema de saúde única.
PALAVRAS-CHAVE: Pets; Epidemiologia; Genes de Resistência; Saúde Única.
PERFIL DE RESISTENCIA EPIDEMIOLÓGICA Y ANTIBIÓTICA Y
DETECCIÓN DE GÉNERO MECA EN STAPHYLOCOCCUS SPP.
ESTUDIANTES VETERINARIOS AISLADOS DE PERROS
RESUMEN: El mantenimiento de las mascotas como reservorios de bacterias
multiresistentes y la transmisión de microorganismos como Staphylococcus spp. entre
animales y seres humanos pueden afectar a la eficacia de los antimicrobianos en la
medicina humana. El objetivo de este estudio fue detectar factores de riesgo, evaluar el
perfil fenotípico de la resistencia antimicrobiana y detectar el gen meccaA en
Staphylococcus spp. aislado de la cavidad nasal de estudiantes veterinarios dueños de
perros. Se trata de una investigación de campo en la que se recogieron 35 muestras de
hisopo nasal para aislar Staphylococcus spp. Se determinó la resistencia antimicrobiana
y la clasificación de los aislados por perfil de multiresistencia (MDR). La presencia del
gen de la mecaA se ha investigado en aislados resistentes a oxacilina. Los estudiantes
respondieron a un cuestionario sobre comportamiento de perros y hábitos de higiene para
identificar variables de riesgo para el desarrollo de resistencia antimicrobiana. Se han
estudiado antimicrobianos como ampicilina, penicilina, oxacilina, cefalotina,

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clindamicina, gentamicina, eritromicina, enrofloxacina y tetraciclina. El 92,9% de los
estafilococos coagulasa-positivos (CoPS) y el 45% de los estafilococos coagulasa-
negativos (CoNS) fueron resistentes a la clase beta-lactam y el 28,6% de los CoPS y el
45% de los CoNS tenían un perfil de MDR. Se clasificaron tres aislados como resistentes
a la oxacilina y se detectó el gen de la mecaA en el 100% de estos aislamientos. Más de
la mitad de los individuos utilizaron antimicrobianos en los últimos 12 meses (52,9%), el
75% utilizó amoxicilina, lo que podría explicar el alto perfil de resistencia antimicrobiana.
Los dueños de perros albergan Staphylococcus spp. con alta resistencia a los
antimicrobianos beta-lactámicos y tienen MDR, lo que representa un único problema de
salud.
PALABRAS CLAVE: Mascotas; Epidemiología; Genes de Resistencia; Salud Única.
1. INTRODUCTION
Currently, the relationship between humans and pets is a driving factor in the
growth of the pet market. Dogs take on a differentiated place since they commonly
become family members in homes, satisfying human needs through their company,
friendship, unconditional love, and affection (PRATA, 2022). However, this very close
relationship between humans and animals can result in the unwanted sharing of
antimicrobial-resistant or multidrug-resistant bacteria.
Staphylococcus spp. are gram-positive bacteria typically found in the upper
respiratory tract and other epithelial surfaces of all warm-blooded animals (MARCO-
FUENTES et al., 2022). However, Staphylococcus aureus, has been associated with
severe infections in humans, such as pneumonia, meningitis, endocarditis, and septicemia
(CHAI et al., 2021). Therefore, it is crucial to determine its resistance profile, mainly
because antimicrobial resistance rates are increasing in both hospital and community
settings, creating a favorable environment for the development of superbacteria (KLOS
et al., 2023). The World Health Organization (WHO) has presented data with high levels
of antimicrobial resistance for several bacterial infections in low-income and high-income
countries, with S. aureus being one of the most commonly reported resistant bacteria
(OPAS, 2018).
Studies evaluating the bacterial phenotypic resistance profile of isolates from
different diseases of dogs and cats (ISHII et al., 2011; SILVA et al., 2014; OLIVEIRA et
al., 2016; MARCO-FUENTES et al., 2022) have shown strains with resistance to most
antimicrobials and many with a multidrug resistance profile. These results show the
possibility of maintenance of multidrug-resistant bacteria and subsequent transmission of
these microorganisms, such as Staphylococcus spp. between animals and humans and

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consequently, an impact on the effectiveness of these drugs in human medicine (LLOYD,
2007; YADAV; KAPLEY, 2021).
Coelho et al. (2007) reported that combining phenotypic and genotypic methods
in identifying the antimicrobial resistance profile of Staphylococcus spp. of humans and
animals allows a more accurate diagnosis and assists in establishing strategies for
controlling the spread of resistant strains.
In bacteria of the genus Staphylococcus, resistance to antimicrobials, especially
beta-lactams, may be associated with producing a penicillin-binding protein (PBP2a or
PBP2’) encoded by the mecA gene (MENDONÇA et al., 2012) and whose expression is
constitutive or induced by beta-lactam antimicrobials such as oxacillin (LOWY, 2003).
Therefore, this study aimed to detect risk factors, evaluate the phenotypic
antimicrobial resistance profile and detect the mecA gene in Staphylococcus spp. isolated
from nasal swabs from dog-owning veterinary medicine students.
2. MATERIAL AND METHODS
2.1 Study Location
Sample collection was conducted in the municipality of Umuarama, located at
23º45'59"S and 53º19'30"W, in northwestern Paraná state, Brazil. Nasal swab samples
were collected from dog-owning veterinary medicine students from a private University
after approval by the Research Ethics Committee Involving Human Beings (CEPEH)
under CAAE No. 71715417.5.0000.0109.
Initially, students were informed about the research project and answered an
epidemiologic questionnaire containing questions about using antimicrobials in the last
12 months and hygienic habits. None of the students showed clinical signs of any illness.
2.2 Sampling
The samples were collected through a nasal swab from the students, considering
the total number of veterinary medicine students (N = 181) in 2017 and the formula for
determining the number of samples to obtain data for statistical analysis (RODRIGUES,
2002):
n0 = 
󰇛󰇜 and n = 



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Where,
n0 - initial number; z - confidence level; p - value obtained from a previous study by others or an origin not
known, is considered 50%; N - population size; q = 100 – p; P – p = precision determined by the researcher
(15%).
Thus, 35 nasal swab samples were collected from the veterinary students. For
sample collection, the swab was first moistened in the transport medium (Stuart). It was
then introduced into the student’s nose, with rotation, and inserted into the transport
medium for further laboratory processing.
2.3 Isolation of Coagulase-Positive Staphylococci (CoPS) and Coagulase-Negative
Staphylococci (CoNS)
The swab was processed at the Laboratory of Preventive Veterinary Medicine and
Public Health of the Graduate Program in Animal Science with an Emphasis on Bioactive
Products at Parana University (UNIPAR). The swab was removed from the transport
medium and incubated in brain heart infusion medium (BHI) in an oven for 24 hours at
37 ºC. Afterward, the bacteria were streaked on mannitol agar and incubated at 37 ºC for
24 hours to isolate Staphylococcus spp. Each colony was submitted to analysis of the
macroscopic, microscopic, catalase, and coagulase tests, allowing classification of
coagulase-positive Staphylococci (CoPS) and coagulase-negative Staphylococci (CoNS).
2.4 Diffusion Susceptibility Test
The agar disc-diffusion method was used according to the recommendations of
the Clinical and Laboratory Standards Institute (CLSI, 2018). The Staphylococcus spp.
were tested for susceptibility to 10 antimicrobials: ampicillin (10 µg), penicillin (10 U),
oxacillin (1 µg), cephalotin (30 µg), cefoxitin (30 µg), clindamycin (2 µg), gentamicin
(10 µg), erythromycin (15 µg), enrofloxacin (5 µg) and tetracycline (30 µg).
Inhibition zone results were subsequently interpreted as sensitive (S) and resistant
(R) according to the reference standard by Br-CAST (BrCAST, 2022), except for
enrofloxacin which was interpreted according to CLSI (CLSI, 2018).
2.5 Detection of the Meca Gene
The samples that were resistant to oxacillin had their DNA extracted with the aid
of a commercial kit (Purelink Genomic DNA Kit - Invitrogen, Carlsbad, CA, USA), for

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carrying out PCR assays, according to Murakami et al. (1991) using the primers mecA1
(AAAATCGATGGTAAAGGTTGG) and mecA2 (AGTTCTGCAGTACCGGATTTG)
at 5 µM. A Veriti™ 96-Well Thermal Cycler Applied Biosystems was used for PCR
amplification. The amplification products were visualized by electrophoresis on 2%
agarose gel stained with Gel Red (Uniscience, Osasco, São Paulo, BR), yielding a 533-
bp band.
2.6 Determination of the Multidrug Resistance Profile - Multidrug Resistance
(MDR)
Bacterial isolates were classified concerning their multidrug resistance (MDR)
profile, defining MDR as not susceptible to at least one agent in at least three classes of
antimicrobials (SWEENEY et al., 2018).
2.7 Statistical Analysis
The association between the variables (hygienic habits) and antimicrobial
resistance profile or multidrug resistance of CoPS and CoNS was determined. The data
were compared using the chi-square test with Yates correction or Fisher's exact test and
calculating the odds ratio and 95% confidence interval using the statistical program SPSS
v. 21.0 at the 5% significance level.
3. RESULTS
Thirty-five isolates of Staphylococcus spp. were obtained. Fourteen were
identified as coagulase-positive staphylococci (CoPS) (Table 1) and 92.9% were resistant
to the beta-lactams and macrolides antimicrobials. In addition, 28.6% presented a multi-
resistance profile. The antimicrobials with the highest resistance profile were penicillin
(PEN) and erythromycin (ERI), resistant to 14 strains of CoPS.
Table 1 - Rate of antimicrobials resistance of coagulase-positive Staphylococci (CoPS) strains (n=14)
isolated from nasal swabs of veterinary medicine students dog owners.
ID
Course series
Antimicrobials to which the strain was resistant
MDR
1
2 nd serie
PEN, ERI
No
5
2 nd serie
PEN, ERI
No
9
2 nd serie
PEN, ERI
No
19
1 st serie
PEN, ERI
No
20
1 st serie
PEN, ERI, GEN
Yes
24
3 rd serie
OXA, PEN, ERI, GEN, TET
Yes
29
4 th serie
PEN, ERI
No
37
4 th serie
PEN, ERI, GEN, TET
Yes

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38
4 th serie
PEN, ERI, TET
No
39
4 th serie
PEN, ERI
No
41
3 rd serie
PEN, ERI, TET
Yes
42
3 rd serie
AMP, PEN, ERI
No
43
3 rd serie
AMP, PEN, ERI
No
44
3 rd serie
AMP, PEN, ERI, TET
Yes
TOTAL
2 -1 st serie
3 -2 nd serie
5 -3 rd serie
4 -4 th serie
Beta-lactams (13/14; 92.9%)
Quinolones (0/14; 0%)
Macrolides (13/14; 92.9%)
Lincosamides (0/14; 0%)
Aminoglycosides (4/14; 28.6%)
Tetracycline (4/14; 28.6%)
MDR
(4/14; 28.6%)
MDR – Multidrug resistance; Beta-lactams class: AMP: Ampicillin; OXA: Oxacillin; PEN: Penicillin;
CFL: Cephalothin; CFO: Cefoxitin. Quinolones: ENO: Enrofloxacin. Macrolides: ERI: Erythromycin.
Lincosamides: CLI: Clindamycin. Aminoglycosides: GEN: Gentamicin. Tetrakinins: TET: Tetracycline
Source: Prepared by the authors.
The 20 strains of CoNS identified (Table 2) presented a lower resistance profile.
That is, 45% were resistant to the beta-lactam and 40% to the macrolide antimicrobials.
However, they showed a higher multidrug resistance profile (45%), emphasizing the same
antimicrobials.
Table 2 - Rate of antimicrobials resistance of coagulase-negative Staphylococci (CoNS) strains (n=20)
isolated from nasal swabs of veterinary medicine students dog owners.
ID
Course series
Antimicrobials to which the strain was resistant
MDR
2
2 nd serie
PEN, ERI, GEN, TET
Yes
3
2 nd serie
PEN, ERI, TET
Yes
4
2 nd serie
AMP, PEN, ERI, GEN, TET, CFO
Yes
7
2 nd serie
PEN, ERI
No
14
1 st serie
AMP, PEN, CFO
No
15
1 st serie
PEN, TET, CFO
No
16
1 st serie
PEN, GEN, TET, CFO
Yes
17
1 st serie
PEN, CLI, ERI, CFO
Yes
18
1 st serie
PEN, CFO
No
21
1 st serie
PEN, ENO, ERI, GEN, TET, CFO
Yes
23
3 rd serie
PEN, ERI, GEN, TET, CFO
Yes
25
3 rd serie
AMP, OXA , PEN, ENO, ERI, GEN, TET, CFO
Yes
30
4 th serie
PEN, ERI
No
31
4 th serie
PEN, ERI
No
32
4 th serie
PEN
No
33
4 th serie
PEN, ERI
No
34
4 th serie
OXA, PEN, GEN
No
35
4 th serie
PEN, ERI, TET, CFO
Yes
36
4 th serie
PEN, ERI, CFO
No
40
4 th serie
PEN, CFL
No
TOTAL
6 -1 st Serie
4 -2 nd Serie
2 -3 rd Serie
8 -4 th Serie
Beta-lactams (9/20; 45%)
Quinolones (2/20; 10%)
Macrolides (8/20; 40%)
Lincosamides (1/20; 5%)
Aminoglycosides (7/20; 35%)
Tetracycline (9/20; 45%)
MDR
(9/20; 45%)
MDR – Multidrug resistance; Beta-lactams class: AMP: Ampicillin; OXA: Oxacillin; PEN: Penicillin;
CFL: Cephalothin; CFO: Cefoxitin. Quinolones: ENO: Enrofloxacin. Macrolides: ERI: Erythromycin.
Lincosamides: CLI: Clindamycin. Aminoglycosides: GEN: Gentamicin. Tetrakinins: TET: Tetracycline.
Source: Prepared by the authors.

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Concerning the use of antimicrobials in the last 12 months, 18 among 34 students
(52.9%) (Table 3) reported the use of antimicrobials, and 12 (75%) used a beta-lactam
antimicrobial (amoxicillin). One of the students did not answer this question.
Table 3 - Absolute (n) and relative (%) behavior of dog-owning veterinary medicine students (n = 34) in
relation to the use of antimicrobials, hygiene habits and habits related to contact with animals.
Variable
n (%)
No
Yes
Did you use an antibiotic in the last 12 months?
16 (47.1%)
18 (52.9%)
Do you wash your hands before eating?
19 (55.9%)
15 (44.1%)
Are you in the habit of washing your hands after contact with animals?
20 (58.8%)
14 (41.2%)
Does the animal have access to outside?
14 (41.2%)
20 (58.8%)
Are you in the habit of sleeping with your pet?
22 (64.7%)
12 (35.3%)
Do you use PPE during a procedure with animals?
---
34 (100%)
PPE: personal protective equipment
Source: Prepared by the authors.
There was no association (P>0.05) between the presence of CoPS and CoNS
strains with a multi-resistance profile and the variables related to the hygiene habits and
behavior of students with their pets (Table 4).
In the molecular tests, the three samples classified as resistant to oxacillin and
subjected to PCR were positive for the mecA gene.
Table 4 - Pet and hygiene habits of dog-owning veterinary medicine students and their relationship with
multidrug resistance profile resistance.
Variable
Answer
CoPS
PMR
P value
CoNS
PMR
P value*
Did you use an antibiotic in the
last 12 months?
No
Yes
2/9 (22.22%)
2/5 (40.0%)
0.5840
3/11(27.27%)
5/8 (62.5%)
0.1809
Do you wash your hands before
eating?
No
Always
1/2 (50.0%)
0/4 (0%)
0.4909
1/1 (100%)
3/8 (37.5%)
1.000
Sometimes
3/8 (37.5%)
4/9 (44.44%)
Are you in the habit of washing
your hands after contact with your
pet?
No
Yes
4/8 (50.0%)
0/6 (0%)
0.0849
5/10 (50.0%)
3/9 (33.3%)
0.6499
Are you in the habit of sleeping
with your pet?
No
Yes
3/8 (37.5%)
1/6 (16.7%)
0.5804
5/13(38.5%)
3/6 (50.0%)
1.000
Do you use PPE during a
procedure with animals?
No
Yes
2/9 (22.2%)
2/5 (40.0%)
0.5804
7/13(53.8%)
1/6 (16.7%)
0.1770
Questions with different numbers, not answered by the student; *Fisher’s exact test. PPE: personal
protective equipment.
Source: Prepared by the authors.
4. DISCUSSION
The results revealed high antimicrobial resistance profile, mainly for beta-lactam
(penicillin) and macrolides (erythromycin) (Tables 1 and 2). The high resistance profile
may be associated with using antimicrobials in the last 12 months by 52.9% of the

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students (Table 3), mainly amoxicillin (75%). According to Yadav and Kapley (2021),
one of the factors associated with the emergence of antimicrobial resistance is related to
antibiotic overuse. Klein et al. (2021) reported that from 2000 to 2015, the sale of
medicines in 76 countries revealed a 65% increase in the use of antimicrobials.
Homologous genes found in pathogens, normal microbiota, and soil
(WOOLHOUSE et al., 2015) indicate that the sharing of resistance between bacteria from
animals, humans, and the environment may justify the increase in bacterial strains with
profiles of resistance and multi-resistance, including those associated with the
consumption of food contaminated with resistant bacteria (SERGELIDIS; ANGELIDIS,
2017).
Amoxicillin is the most prescribed antimicrobial in Brazil (LIMA et al., 2014;
PAULA, 2014). In addition, Silva et al. (2013) reported that more than 70% of S. aureus
(CoPS) and S. epidermidis (CoNS) isolates from the community or the hospital
environment are resistant to penicillin, ampicillin and amoxicillin, corroborating the
results found in the present study for penicillin.
Dobrachinski et al. (2022), studying employees of a Public Health unit in Bahia
state, found that all samples of S. aureus (CoPS) isolated from the employees' nasal
cavities were resistant to penicillin, as was observed in this study, in which 100% of the
CoPS strains were resistant to penicillin.
Lin et al. (2018) verified that school environmental contamination with
methicillin-resistant Staphylococcus aureus (MRSA) was positively associated with
MRSA nasal carriage in elementary students. Although the present study did not assess
MRSA strains, the fact that environmental contamination influenced the contamination
of students demonstrates the sharing of antimicrobial-resistant microbiota.
Coelho et al. (2007), evaluating the resistance profile of Staphylococcus spp. from
animals and humans, also observed a high resistance profile for antimicrobials of the beta-
lactam class (ampicillin and penicillin), with a higher percentage for human samples
(80.9%). Penicillin is among the most used antimicrobials in the small animal clinic
(ARIAS; CARRILHO, 2012), which could explain the high antimicrobial resistance in
this drug class in the Staphylococcus isolates tested.
Additionally, we highlight that the students of veterinary medicine subjects of this
research conduct their practical activities with animals, including dogs, and can share
resistant microorganisms with the animals. Coincidently, amoxicillin is one of the main

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antimicrobials prescribed in the small animal clinic for infections by gram-positive
bacteria at the veterinary hospital where the students perform practical activities. In
addition, the use of the combination of amoxicillin with clavulanate for respiratory
infections and cephalexin for pets' skin infections caused by gram-positive bacteria was
also mentioned by veterinarians working at that hospital.
Pereira and Cunha (2009) evaluated nasal colonization by Staphylococcus spp.
resistant to oxacillin in nursing students and similarly observed a low profile of oxacillin
resistance. However, resistance was only detected in CoNS (16.2%), differing from the
present work in which an isolate resistant to oxacillin and with the mecA gene in CoPS
was found.
The occurrence of the mecA gene was evaluated only in samples resistant to
oxacillin, and the results showed that 100% (n=3) had the mecA gene. Similarly, Angel
et al. (2019) found that of the 32 oxacillin-resistant S. aureus isolated from patients in
two hospitals, 30 (93.8%) harbored the mecA gene. Mimica and Mendes (2007) reported
that phenotypic resistance to oxacillin is highly variable and dependent on the expression
of the mecA gene. However, Goering et al. (2019) reported that strains of S. aureus
possessing the mecA gene but showing phenotypic susceptibility to oxacillin may become
resistant after exposure to the antimicrobial, resulting in treatment failure.
In the present study, when the habits of dog-owning students (Table 3) were
related to MDR profile, there was no significant association (P>0.05) between habits
concerning hygiene and behavior with animals and MDR profile. However, this
demonstrates that at some point in life, they were exposed to or showed habits explaining
the high percentage of resistance to antimicrobials of the beta-lactam class. In addition,
in analyzing the questionnaire answers of students about hygiene habits and habits related
to contact with animals (Table 3), 100% made use of personal protective equipment
(mainly gloves) during practical activities with animals, and only 35.3% had the habit of
sleeping with their pet.
Among the CoPS isolated, 28.6% exhibited an MDR profile, similar to the data
from Angel et al. (2019) in S. aureus isolates from urine, vagina, ear, and wound samples
from patients in two hospitals.
Staphylococcus spp. do not live isolated on the skin or mucous surfaces of animals
but are in very close contact with various other bacteria (WENDLANDT et al., 2013),
enabling the sharing of resistance genes. In addition, the interruption of antimicrobial

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treatment when the patient sees improvement allows some bacteria to survive, which
results in the recurrence of the disease, contributing to the growth of bacteria that
survived, possibly including drug-resistant populations (ALLEL et al., 2023).
Strategies aimed at combating resistant pathogens are frequently based on
international documents, such as from the FDA (Food and Drug Administration) and
CDC (Centers for Disease Control and Prevention), that consider many resources and
disease conditions not present in Brazil (COSTA; SILVA JÚNIOR, 2017). These
hindrances and the fact that treatments guided by antibiogram results are rare limit the
measures for control and prevention in Brazil (MENEZES et al., 2006). Since the launch
by WHO of the Global Action Plan on Antimicrobial Resistance in 2015 (ANVISA,
2018), the Brazilian government developed the National Plan for the Prevention and
Control of Resistance to Antimicrobials in the Scope of One Health in 2018 (BRASIL,
2018), which includes, among other objectives, the training of professional managers who
work in the areas of human, animal and environmental health concerning antimicrobial
resistance, demonstrating the great importance of the theme.
5. CONCLUSIONS
Dog-owning students host Staphylococcus spp. with a high antibiotic resistance
profile, mostly drugs of the beta-lactam class. In addition, the Staphylococcus isolates
show a multidrug resistance profile, with the mecA gene present in 100% of the isolates
resistant to oxacillin, proving to be an One health problem, as it may have been acquired
from animals in your household. Further work is needed to verify the association between
the resistance profile in strains of Staphylococcus spp. isolated from dogs and their
respective owners.

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