Open Veterinary Journal, (2025), Vol. 15(9): 4075-4081

Short Communication

10.5455/OVJ.2025.v15.i9.11

Antimicrobial resistance of coagulase-negative Staphylococcus isolated from the external ear canal of healthy cats in Talca, Chile, South América preliminary results

Andrea Núñez^1*, Rodrigo Castro^2,* and Pamela Thomson^3,4

¹Escuela de Medicina Veterinaria, Facultad de Ciencias Agrarias y Forestales, Universidad Católica del Maule, Curicó, Chile

²Escuela de Medicina Veterinaria, Facultad de Recursos Naturales y Medicina Veterinaria, Universidad Santo Tomás, Chile

³Escuela de Medicina Veterinaria, Facultad de Ciencias de la Vida, Universidad Andrés Bello, Santiago, Chile

⁴One Health Institute, Faculty of Life Sciences, Universidad Andrés Bello, Santiago, Chile

*Corresponding Author: Andrea Núñez Bustamante. Escuela de Medicina Veterinaria, Facultad de Ciencias Agrarias y Forestales, Universidad Católica del Maule, Curicó, Chile. Email: anunezb [at] ucm.cl; Rodrigo Castro, Escuela de Medicina Veterinaria, Facultad de Recursos Naturales y Medicina Veterinaria, Universidad Santo Tomás, Chile. Email: rodrigocastro [at] santotomas.cl

Submitted: 15/05/2025 Revised: 05/08/2025 Accepted: 16/08/2025 Published: 30/09/2025

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ABSTRACT

Background: The microbiota of the external ear of cats comprises different types of bacteria that can change due to intrinsic or extrinsic factors. Coagulase-negative Staphylococcus (CoNS) is predominant in the external ear of cats. Antibiotic-resistant CoNS associated with infectious pathologies has been reported in humans. The presence of antibiotic-resistant CoNS acting as reservoirs in the ears of healthy cats could represent a zoonotic risk. This has not yet been studied in Talca, Chile.

Aim: This study aimed to determine the antimicrobial susceptibility of CoNS strains obtained from the external ear of healthy cats in Talca, Chile, as a possible zoonotic risk.

Methods: Ear swabs were obtained from 31 healthy cats. Bacterial culture and morphological analysis were performed on the samples. MALDI-TOF was used to identify CoNS strains, which were tested against a panel of 11 antibiotics using the Kirby-Bauer disk diffusion method. Fisher’s exact test was used to determine the association between the epidemiological variables of the patients and the antimicrobial susceptibility of CoNS strains.

Results: Nine CoNS strains were isolated from the ear of seven healthy cats (prevalence 22.58%). Staphylococcus simulans and Staphylococcus felis were the main species identified. Antimicrobial resistance was observed in 55.5% of the strains, mainly to oxacillin. Resistance to more than two antibiotics was observed in 22.2% of the strains. No association was found between the epidemiological variables of the patients and the antimicrobial susceptibility of the CoNS strains.

Conclusion: The ears of healthy cats can act as a reservoir for antibiotic-resistant CoNS, with a potential risk of transmission to humans. These preliminary results are the first to be reported in Talca, Chile.

Keywords: Staphylococcus, Healthy cat, Ear, Antimicrobial resistance.

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Introduction

Staphylococcus spp. is a diverse genus of facultative anaerobic Gram-positive bacteria comprising more than 40 different species. These bacteria are commensals of the skin and mucous membranes in humans and animals and can cause serious infections in different tissues (Morris et al., 2017). Coagulase-positive Staphylococcus species are described as the agents most associated with infections in both animals and humans, whereas coagulase-negative Staphylococcus (CoNS) is reported as part of the skin and mucosal microbiota (Weese, 2013; Becker et al., 2014; Possa de Menezes et al., 2025). However, the CoNS group has currently been described as having an important clinical relevance due to its virulence and pathogenicity factors (Elnageh et al., 2021; Sukur and Esendal 2020; Franca et al., 2021). CoNS have been associated with serious infections in humans, mainly as opportunistic agents in immunocompromised patients (Asante et al., 2020) and as nosocomial pathogens associated with the use of invasive medical devices (Becker et al., 2014). In cats, they are part of the skin and ear microbiota and have been described in lower urinary tract infections, eye infections, and otitis (Litster et al., 2007; Loeffler et al., 2011; Nocera et al., 2021), with Staphylococcus felis, Staphylococcus simulans, and Staphylococcus schleiferi being the most described species (Ma et al., 2020).

Currently, these species constitute a therapeutic challenge (Becker et al., 2014), as methicillin-resistant isolates and others with resistance to more than two antimicrobials have been reported (Hiramatsu et al., 2001; Suepaul et al., 2021; Miszczak et al., 2023; Temmerman et al., 2024). This study aimed to determine the antimicrobial susceptibility of CoNS strains obtained from the external ear canal of healthy cats.

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Materials and Methods

From December 2021 to March 2022, a total of 31 healthy cats were sampled and attended for routine check-ups in the city of Talca, Chile (35°26′00”S; 71°40′00”W), who underwent a history and physical examination under the management of “cat friendly” care, recording epidemiological variables and variables of clinical interest, such as: age (kitten=birth up to 1 year; young adult=1–6 years; mature adult=7–10 years; senior > 10 years) (Quimby et al., 2021); sex (male or female); reproductive status (whole; sterilized); breed Domestic Short Haired (DSH); Domestic Long Haired (DLH); confinement (indoor or outdoor); having been hospitalized at some point in life (yes or not); has received antibiotic treatment at some point in life (yes or not).

Exclusion criteria

Cats with the following clinical records were excluded from this study: cats with clinical otitis; cats that have been hospitalized in the last 30 days prior to sampling; cats that have received antibiotic treatment in the last 30 days prior to sampling.

Sampling

Samples were taken from the right external auditory canal (Older et al., 2021) using a sterile swab in Stuart transport medium (Linsan, Santiago, Chile) and transported immediately to the Clinical Microbiology Laboratory of the Andrés Bello University, Metropolitan Region.

Sample processing

Each swab was seeded on mannitol salt agar (Becton Dickinson, Heidelberg, Germany) and incubated at 37°C for 24 hours. Then, the different cultured morphotypes were evaluated semi-quantitatively. Gram-positive, catalase-positive, and coagulase-negative morphotypes (BD BBL™) they were isolated in blood agar (Linsan, Santiago, Chile) and subsequently identified by MALDI-TOF mass spectrometry (Maldi Biotyper, Bruker, Billerica, MA, USA) (Oliveira et al., 2021). All CoNS isolates were tested against a panel of 11 antibiotics using the Kirby-Bauer disk diffusion method following the CLSI guidelines in document VET01S (CLSI, 2020). The antibiotics tested included cefoxitin (FOX, 30 μg), oxacillin (OX, 1 μg), imipenem (IPM, 10 μg), ciprofloxacin (CIP, 5 μg), doxycycline (OD, 30 μg), erythromycin (E, 15 μg), amikacin (AMK, 30 μg), gentamicin (GEN, 10 μg), trimethoprim/sulfamethoxazole (SXT, 1.25/23.75 μg), amoxicillin/clavulanic acid (AMC, 30 μg), and clindamycin (DA, 2 μg), all of which were supplied by OXOID (Hampshire, UK). The methicillin-resistant phenotype was evaluated using an OX disc. The reference strain was Staphylococcus aureus ATCC 25923. The strains were classified into three categories according to zone diameter breakpoints (mm): sensitive, intermediate sensitive, or antibiotic resistant (Table 1). Bacterial isolates resistant to three or more classes of antimicrobials were classified as multidrug resistance (MDR) according to previously standardized criteria (Magiorakos et al., 2012).

Table 1. Interpretative categories and zone diameter breakpoints (mm) for each antibiotic studied.

Statistical analysis

Descriptive statistics were used to characterize the distribution of the isolated strains and their antimicrobial susceptibility. The association between the epidemiological variables of the patients and the antimicrobial susceptibility of the Staphylococcus strainswas determined using the exact Fisher test, with 95% confidence and a α error of 5% (Di Rienzo et al., 2008).

Ethical approval

The study received the approval of the Scientific Ethics Committee of the South Central Macrozone of the Santo Tomás University (CEC-CS UST), code n°129-21.

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Results

Of the 31 healthy cats studied, CoNS was detected in seven of them (prevalence 22.58%). The most frequent variables were: age kitten (45.16%), male sex (58.06%), breed DSH (80.65%), reproductive status (sterilized) (58.06%), and confinement indoor (64.52%). On the other hand, 67.64% of cats have not been hospitalized, and 61.29% have received antibiotic treatment at some point in their lives (Table 2). No association was found between epidemiological variables and Staphylococcus isolation (p > 0.05).

Table 2. Absolute (n) and relative (%) frequency distribution of the epidemiological variables and detection of strains CoNS in the 31 patients included in the study.

Nine CoNS strains were isolated and identified as follows: 3 S. simulans, 2 S. felis, 1 S. capitis, 1 S. epidermidis, 1 S. pettenkoferi, and 1 S. hominis (Table 3). Although the nine strains were sensitive to the antimicrobials gentamicin, imipenem, doxycycline, cefoxitin, amoxicillin, clavulanic acid, amikacin, and enrofloxacin, 55.5% of the strains presented resistance to other antimicrobials, with oxacillin being the most frequent, followed by erythromycin. One strain of S. felis presented with MDR to clindamycin, erythromycin, oxacillin, and sulfa-trimethoprim (Table 3). The strains that presented methicillin resistance (MRS) were S. capitis, S. epidermidis, S. pettenkoferi, S. simulans, and S. felis.

Table 3. Description of feline patients according to epidemiological variables, species and antimicrobial susceptibility of the strains obtained.

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Discussion

Companion animals are colonized by various Staphylococcus spp. populations (Kaspar et al., 2018). However, the proportion of coagulase-negative species and their relative distribution vary between studies, possibly due to the different anatomical sites chosen for sampling (Weese, 2013). Thus, Ma et al. (2020) reported a high colonization rate of various species, particularly CoNS, which was significantly higher in dogs than in cats.

In the group of 31 cats studied in this study, the ears of healthy cats can act as a reservoir of antibiotic-resistant CoNS, with a potential risk of transmission to humans. These preliminary results are the first reported in Talca, Chile, and not only provide prevalence data but also allow us to identify patterns of antimicrobial resistence (AMR), which should be considered in future lines of research aimed at epidemiological surveillance and genetic characterization of resistance genes. This local information provides background information that contributes to a better understanding of the global phenomenon of AMR and public health and may be useful for other locations in the world that share similar conditions. Talca, Chile (35°25ʹS, 71°39ʹW), is a rural South American city characterized by a temperate Mediterranean climate, with an average rainfall of 475 and 29 mm in winter and summer, respectively, and an average temperature of 18.8°C (Thomson et al., 2022a).

In the present study, all strains obtained were identified as CoNS, and S. simulans was the most frequently isolated strain (33.3%), which is similar to the results reported by Elnageh et al. (2021), who obtained 63% of CoNS group strains isolated mainly from healthy cats. Staphylococcus simulans is an emerging cutaneous pathogen, whose zoonotic potential has already been documented and isolated from human infections (Shields et al., 2016). The second most frequently isolated strain was S. felis (22.2%), which possesses virulence genes capable of generating pathologies, mainly associated with urinary tract infections and otitis externa (Worthing et al., 2018). In a previous study conducted in our country, the most frequently isolated species in animals and humans was S. felis (Thomson et al., 2022b).

The strains that presented MRS were S. capitis, S. epidermidis, S. pettenkoferi, S. simulans, and S. felis. The latter was also MDR and came from a 5-month-old, indoor, whole female patient who was not previously hospitalized but did receive antimicrobial therapy. Although S. felis is a CoNS species usually isolated from the ear of healthy cats, there is evidence of its zoonotic potential causing infection in human skin wounds (Sips et al., 2023). Therefore, the MDR condition recognized in this study should be considered for epidemiological follow-up studies because this strain was isolated from a healthy indoor kitten. Incorrect antibiotic prescribing practices or indications could contribute to AMR (King et al., 2018). In the present study, 6 of the 7 cats carrying CoNS had received antibiotic treatment, a condition associated with the resistance profile observed in the strains (Jung et al., 2020). In Chile, Galarce et al. (2021) surveyed veterinarians regarding the use of antimicrobials for treating bacterial diseases and the use of laboratory diagnostic tools. Thus, the most commonly used antimicrobials by professionals were beta-lactams (65.3%), quinolones (36.2%), and tetracyclines (23.2%) (Galarce et al., 2021). Although 67% stated that they were aware of the official antimicrobial classification schemes, only 15% of veterinarians requested laboratory diagnostic tests for such a prescription. With these results, the authors describe the need for official efforts aimed at reducing the widespread use of antimicrobials in companion animals, promoting the correct prescription of antimicrobials, and establishing protocols for the use of antimicrobials, given the importance of working under the One Health approach.

In this study, no S. aureus strains were identified, which is not related to what was obtained by other authors, who described that the colonization of this species in healthy cats is frequent (De Jong et al., 2020). This difference could be attributed to the larger sample size used at various anatomical sites.

Changes in the ear microclimate, due to immune-based pathologies, can induce the excessive growth of bacteria and other microorganisms, predisposing animals to bacterial otitis (Coutinho et al., 2006), and companion animals can harbor MRS strains, which represent a risk to public health. Thus, the reported antimicrobial resistance among CoNS strains requires greater attention and continuous monitoring by veterinarians and public health professionals (Elnageh et al., 2021). Risk factors for infections by these resistant strains in dogs and cats include previous hospitalization, frequent visits to veterinary clinics, glucocorticoid administration, and antibiotic treatment (Lehner et al., 2014; Krapf et al., 2019). Cats with a history of antimicrobial treatment and hospital stay are at greater risk of acquiring MRS strains (Souza et al., 2024). Different environments, surfaces, and equipment of veterinary clinics can be contaminated with bacteria resistant to multiple antimicrobials (Espadale et al., 2018). Community colonization of these strains is a risk factor for cross-transmission of resistant bacteria between cats and humans. Therefore, the emergence and spread of AMR in cats represents a challenge in veterinary medicine, both from the point of view of patient health and public health. According to the One Health perspective, it is relevant to carry out epidemiological surveillance and investigate the role that companion animals play in the transmission of these pathogens to better understand the mechanism of transmission of antimicrobial resistance between veterinary medicine and humans (Favier et al., 2018; Jung et al., 2020).

In the present study, there was no association between epidemiological variables and CoNS isolation (p > 0.05). However, the lack of significant associations with other epidemiological variables could reflect the independence of the condition with respect to these factors or could be due to limitations in the study sample size, in particular the small group of only seven cats with CoNS isolation, which could reduce the study’s ability to detect small effects and limit the generalizability of the findings.

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Conclusion

The ears of healthy cats can act as a reservoir for antibiotic-resistant CoNS, with a potential risk of transmission to humans. These preliminary results are the first reported in Talca, Chile, and suggest that future follow-up studies, such as epidemiological surveillance and genetic characterization of resistance genes, should be conducted.

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Acknowledgments

The authors would like to thank the veterinarians and veterinary clinics of the city of Talca for their invaluable help in providing patients for the realization of this study.

Conflict of interest

The authors declare that there is no conflict of interest.

Funding

None.

Authors’ contributions

Andrea Núñez: Conception and design of the study, wrote the first draft of the manuscript, critically revised the manuscript, and methodology. Pamela Thomson: writing the first draft of the manuscript and methodology. Rodrigo Castro: Writing on one of the topics, statistical methodology, and critically revising the manuscript. All authors have read and agreed to the published version of the manuscript.

Data availability

All data were provided in the manuscript.

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