Next Article in Journal
Beyond Penicillin: The Potential of Filamentous Fungi for Drug Discovery in the Age of Antibiotic Resistance
Next Article in Special Issue
Susceptibility of Actinobacillus pleuropneumoniae, Pasteurella multocida and Streptococcus suis Isolated from Pigs in Hungary between 2018 and 2021
Previous Article in Journal
Antimicrobial Resistance, Genetic Lineages, and Biofilm Formation in Pseudomonas aeruginosa Isolated from Human Infections: An Emerging One Health Concern
Previous Article in Special Issue
Occurrence of Antimicrobial Resistance in Canine and Feline Bacterial Pathogens in Germany under the Impact of the TÄHAV Amendment in 2018
 
 
Search for Articles:
Title / Keyword
Author / Affiliation / Email
Journal
Article Type
 
 
Advanced Search
 
Section
Special Issue
Volume
Issue
Number
Page
 
Journals
Antibiotics
Volume 12
Issue 8
10.3390/antibiotics12081249
antibiotics-logo
Submit to this Journal Review for this Journal Propose a Special Issue
► Article Menu

Article Menu

share Share announcement Help format_quote Cite question_answer Discuss in SciProfiles thumb_up
...
Endorse textsms
...
Comment
first_page
settings
Order Article Reprints
Font Type:
Arial Georgia Verdana
Font Size:
Aa Aa Aa
Line Spacing:
Column Width:
Background:
Article

Fecal Carriage of Extended-Spectrum β-Lactamase-/AmpC-Producing Escherichia coli in Pet and Stray Cats

by
Gabriele Ratti
1,
Alessia Facchin
1,
Angelica Stranieri
1,
Alessia Giordano
1,
Saverio Paltrinieri
1,
Paola Scarpa
1,
Deborah Maragno
1,
Alessia Gazzonis
1,
Martina Penati
1,2,
Camilla Luzzago
1,
Paola Dall’Ara
1,* and
Stefania Lauzi
1
1
Department of Veterinary Medicine and Animal Sciences (DIVAS), University of Milan, 26900 Lodi, Italy
2
Laboratory of Animal Infectious Diseases (MiLab), University of Milan, 26900 Lodi, Italy
*
Author to whom correspondence should be addressed.
Antibiotics 2023, 12(8), 1249; https://doi.org/10.3390/antibiotics12081249
Submission received: 5 July 2023 / Revised: 21 July 2023 / Accepted: 26 July 2023 / Published: 29 July 2023
(This article belongs to the Special Issue Epidemiology, Impact and Mitigation of Antimicrobial Resistance in Veterinary Medicine, 2nd Edition)
Review Reports Versions Notes

Abstract

:
Dogs have been reported as potential carriers of antimicrobial-resistant bacteria, but the role of cats has been poorly studied. The aim of this study was to investigate the presence and the risk factors associated with the fecal carriage of extended-spectrum β-lactamase and AmpC (ESBL/AmpC)-producing Escherichia coli (E. coli) in pet and stray cats. Fecal samples were collected between 2020 and 2022 from healthy and unhealthy cats and screened for ESBL/AmpC-producing E. coli using selective media. The presence of ESBL/AmpC-producing E. coli was confirmed by phenotypic and molecular methods. The evaluation of minimum inhibitory concentrations (MICs) was performed on positive isolates. Host and hospitalization data were analyzed to identify risk factors. A total of 97 cats’ samples were collected, and ESBL/AmpC-producing E. coli were detected in 6/97 (6.2%), supported by the detection of blaCTX-M (100%), blaTEM (83.3%), and blaSHV (16.7%) genes and the overexpression of chromosomal ampC (1%). All E. coli isolates were categorized as multidrug-resistant. Unhealthy status and previous antibiotic therapy were significantly associated with ESBL/AmpC-producing E. coli fecal carriage. Our results suggest that cats may be carriers of ESBL/AmpC-producing E. coli, highlighting the need for antimicrobial stewardship in veterinary medicine and an antimicrobial-resistance surveillance program focusing on companion animals, including stray cats.

1. Introduction

Antimicrobial resistance (AMR) has become one of the major threats to public health [1]. Multisectoral surveillance systems focusing on resistance to antibiotics are key points to define and evaluate the effectiveness of measures against AMR. Due to their rapid emergence over the last few years, concerns have been raised about the spread of extended-spectrum beta-lactamase (ESBL)- and AmpC-producing Escherichia coli (E. coli) in both humans and animals [2,3]. The importance of ESBL/AmpC-producing E. coli strains has been emphasized by their association with increasing treatment failure, hospitalization, and mortality in humans and animals [4]. The need for a One Health approach has been highlighted [5], and ESBL/AmpC-producing E. coli have been proposed as commensal indicator microorganisms for AMR surveillance systems [5,6].
Most of the studies and national surveillance programs related to AMR have focused on food-producing animals [7,8,9]. However, the close contact and sharing of environments between humans and companion animals as well as the wide spread of dogs and cats in European households has raised concerns about the role of companion animals in AMR dissemination and the potential risk of the transmission of resistant bacteria to humans or vice versa [2,10]. The presence of ESBL-producing E. coli has been reported among dogs worldwide, with different prevalence levels likely reflecting differences in the diagnostic methods used; the levels of antibiotic use among veterinarians and owners; and other factors influencing bacterial transmission, including socioeconomic and behavioral components [11]. Similarly to humans and food-producing animals, the most frequent genes associated with ESBL resistance in dogs encode for CTX-M enzymes, followed by TEM and SHV [12,13]. Moreover, the presence of AmpC-producing E. coli in dogs, usually carrying the blaCMY-2 gene, has been reported with a lower prevalence compared to ESBL-producing E. coli [3,13,14].
Despite being the most popular companion animals in Europe, few studies have investigated the presence of ESBL/AmpC-producing E. coli in cats [11,15]. The presence of ESBL/AmpC-producing E. coli in cats has been reported in clinical samples from diseased cats (with a variety of common clinical conditions, including gastrointestinal disease, upper respiratory tract disease, otitis, conjunctivitis, stomatitis, skin abscess, and urinary tract infections) and in fecal samples from healthy cats [3,14,16,17]. In addition to owned cats, stray cats have also been reported as reservoirs of AMR E. coli [18]. The role of host factors associated with the spread of ESBL/AmpC-producing E. coli in cats still needs to be elucidated [2,19]. Given the importance of cats as a potential ESBL/AmpC-producing E. coli reservoir because of their wide diffusion as pet animals and their close contact and sharing of environments with humans, the aims of this study were to estimate the presence of ESBL/AmpC-producing E. coli fecal carriage in cats, characterize the antimicrobial-resistance phenotypes and genotypes of the isolates, and identify risk factors associated with the fecal carriage of ESBL/AmpC-producing E. coli.

2. Results

In total, 97 fecal samples from cats were included in this study. The characteristics of the analyzed cats are summarized in Table 1. Statistical analysis showed that the unhealthy status of animals (OR = 5.91; 95% CI: 1.01–34.44; p = 0.049) and previous antibiotic therapy (OR = 6.67; 95% CI: 1.14–38.99; p = 0.037) were significantly associated with the fecal carriage of ESBL/AmpC-producing E. coli, whereas the other variables considered in the analysis were not risk factors (Table 1). The two positive stray cats detected in this study originated from two different feline colonies.
The characteristics of the unhealthy cats compared to healthy cats analyzed in this study are given in Table S1.
Overall, the presence of ESBL/AmpC-producing E. coli was detected in 6/97 (6.2%; 95% confidence interval: 1.4–11%) cats, and the characteristics of the positive cats are reported in Table 1. More precisely, ESBL-producing E. coli was only observed in five (83.3%) isolates, whereas ESBL- and AmpC-producing E. coli were observed in one (16.7%) strain only. The results of the phenotypic and genetic characterization of the E. coli isolates are summarized in Table 2. Among the blaCTX-M-positive isolates, all E. coli isolates carried the blaCTX-M-1 group gene.
The analysis of the promoter/attenuator region of the AmpC-producing E. coli detected in this study showed the presence of a −32 T > A mutation in the −35 promoter box and a −28 G > A mutation in the spacer region (Table S2), previously associated with the upregulation of AmpC production [20].
Based on MIC results (Table 3), the highest level of resistance was observed for β-lactam, with all isolates resistant to ampicillin, cefazolin, cephalexin, cefovecin, and cefpodoxime, and for doxycycline. Resistance to fluoroquinolones (16.7%); phenicol (16.7%); β-lactam in combination with the β-lactamase inhibitor agent (16.7% and 50% for amoxicillin/clavulanic acid 2:1 and piperacillin/tazobactam content 4, respectively); tetracycline (83.3%); and carbapenems (16.7%) was less frequent, with at least one isolate resistant to all the agents in each category. All isolates were susceptible to amikacin, accounting for the lowest resistance rate. All ESBL/AmpC-producing E. coli isolates detected in this study were MDR.

3. Discussion

Cats are the most common companion animals worldwide, but only a few studies have investigated their role as potential carriers of AMR bacteria [7,11,21]. In the present study, fecal samples were analyzed from stray and pet cats to estimate the presence of ESBL/AmpC-producing E. coli fecal carriage in cats from Italy and the associated risk factors. The overall presence of ESBL/AmpC-producing E. coli in 6.2% of cats detected in this study was consistent with previous reports that have investigated the fecal carriage of ESBL-producing E. coli in healthy and sick cats from Portugal [3] and in sheltered cats [22]. The recent meta-analysis performed by Salgado-Caxito and colleagues [11] showing that the global estimated prevalence of ESBL-producing E. coli in cats is 5.04% (95% CI: 2.42–10.22%) also confirms our results. However, other studies have detected a prevalence of up to 20% for ESBL-producing E. coli among cat fecal samples [23,24], probably due to the different backgrounds of the tested cats or the different diagnostic methods used. Indeed, a comparison between studies may be challenging due to the lack of standardized diagnostic methods for AMR surveillance in pets [11,13]. Therefore, according to standardized protocols for food-producing animals [6], the implementation of a specific standardized diagnostics methodology for AMR surveillance programs in companion animals may be suggested.
The presence of the ESBL genotype supported by the detection of blaCTX-M as the main resistance gene in all isolates in this study, followed by blaTEM and blaSHV, confirmed previous studies reporting these as the most common genes in ESBL-producing bacteria from both humans and animals [25,26], though it should be noted that we did not determine the TEM and SHV groups of the isolates, which do not always encode for ESBL [27]. Our results showing that the majority of E. coli isolates carried more than one resistance gene confirmed the genetic diversity of ESBL-producing E. coli in companion animals [11,13]. Moreover, the high detection rate of blaCTX-M-1 group genes was expected, as this group is the most common worldwide [28].
The presence of mutations of the AmpC-producing E. coli isolates detected in this study associated with the upregulation of AmpC production [20] was consistent with previous reports highlighting the presence of campC resistance in companion animals [29,30]. However, it must be noted that AmpC hyperproduction is less frequently investigated compared to plasmid-mediated AmpC resistance, since the latter mechanism is capable of being horizontally transferred to other bacteria, posing a greater threat to AMR control [20,31]. Indeed, the absence of pampC among the AmpC-producing E. coli detected in this study could have been due to the limited sample size and the lower presence of pAmpC-producing E. coli compared to ESBL-producing E. coli in companion animals [3,13,14,17,32]. Therefore, further studies with a wider sample size are necessary to assess the role of cats in the dissemination of pAmpC-producing E. coli. Moreover, our results confirmed that commercially available kits for AmpC detection failed to differentiate between pAmpC and cAmpC production in E. coli, as previously reported [31].
Interestingly, one isolate detected in this study that carried blaCTX-M and blaTEM genes was phenotypically negative for ESBL production. The ESBL phenotype could have been masked by the presence of AmpC β-lactamases and/or carbapenemases [33]. However, the presence of AmpC β-lactamases was unlikely, as the isolate was negative for the AmpC phenotype. Interestingly, this isolate was the only one resistant to imipenem, with an MIC value ≥16 µg/mL. A high imipenem MIC value has been used as a screening method for carbapenemase production in Enterobacteriaceae [34,35], as isolates with an MIC value of 2 µg/mL or above are likely to be carbapenemase producers [36,37]. Unfortunately, the aim of our study did not include the detection of carbapenemase-producing bacteria. In this respect, further studies are needed to clarify the epidemiological role of cats in the carriage of carbapenemase-producing E. coli, as the presence of carbapenemase-producing Enterobacteriaceae in companion animals has been rarely investigated, despite recent studies having shown infection or colonization by carbapenemase-producing Enterobacterales in companion animals [21,38,39].
Regarding E. coli phylogroups, the detection of B2 as the most frequent phylogenetic group was consistent with previous reports showing a high presence of phylogroup B2 among the AMR E. coli in companion animals and humans but not in food-producing animals, suggesting the need for further investigations to understand the zoonotic potential of these isolates [17,40,41,42,43,44]. Moreover, given that E. coli belonging to phylogroups B2 and F has been commonly associated with the presence of virulence factors responsible for extra-intestinal infections [40,44], further investigations are suggested to assess the virulotyping of ESBL/AmpC-producing E. coli in cats.
The high presence of MDR E. coli was expected, as ESBL-producing E. coli usually not only exhibit resistance to β-lactam, but frequently carry other resistance genes conferring resistance to other antimicrobial drugs, leading to MDR [45]. Moreover, the high detection rate of MDR E. coli highlighted the need for antimicrobial stewardship in veterinary practice to also reduce the emergence and spread of MDR bacteria in cats.
The unhealthy status of cats as a risk factor associated with the fecal carriage of ESBL/AmpC-producing E. coli observed in our study was likely influenced by the fact that a higher percentage of sick animals were treated with antibiotics compared to healthy cats. This suggests that antibiotic treatment alone may have increased the risk of the fecal carriage of AMR organisms in this study, as it is a well-recognized risk factor for the acquisition of drug-resistant bacteria in both humans and animals [2,46,47]. It was not possible to define if specific antibiotic classes were associated with ESBL/AmpC-producing E. coli due to the low number of cats treated with antibiotics in this study. The absence of an association between hospitalization and the fecal carriage of ESBL/AmpC-producing E. coli could have been due to the low number of hospitalized cats included in the present study or to the differences in management and hygiene protocols among hospitals [19], since hospitalization has been associated with the carriage of drug-resistant bacteria in companion animals [19,46,47]. In this respect, further studies focusing on hospitalized cats are needed to evaluate the contribution of veterinary hospitals in the spread of drug-resistant bacteria.
The presence of ESBL/AmpC-producing E. coli among both stray and owned cats is of interest and confirms a recent report detecting AMR in stray cats [18]. Given the unknown backgrounds of the stray cats in this study, we could not conclude if the AMR acquisition was associated with environmental sources, antimicrobial administration, the presence of subclinical disease (cats were healthy at physical examination), or other factors. Moreover, the apparent non-statistically significant higher ESBL/AmpC-producing E. coli carriage in owned cats compared to stray cats may have been influenced by the high percentage of unhealthy animals among the owned cats, likely requiring antimicrobial therapy. Indeed, although this group of cats was raised in an enclosed environment, with a clean water and food supply and a good level of care from their owners, more than half of the owned cats were unhealthy, whereas all stray cats were apparently healthy and were brought to the VTH only for neutering.
Further investigations are needed to understand the potential route of transmission of AMR in stray cat populations as well as to clarify the role of stray cats, as previously reported for stray dogs in South America [14,48].
This study had some limitations, starting from the low number of analyzed cats, which was not sufficient for population estimation. Further studies with a greater sample size and an ad hoc sampling strategy are needed to assess the epidemiological role of stray and owned cats in the spread of ESBL/AmpC-producing E. coli. The whole-genome sequencing of the E. coli isolates detected in this study is suggested in order to define virulence factors and the presence of other resistance genes that were not investigated in this study. Moreover, it would be interesting to analyze more than one AMR isolate from each sample to investigate if the genetic variability of E. coli isolates can occur within the same fecal sample.

4. Materials and Methods

4.1. Sample Collection

Fecal samples were collected from both owned and stray cats admitted to the Veterinary Teaching Hospital (VTH) of Lodi, University of Milan, Italy from 2020 to 2022. Fecal samples from owned cats were collected from leftover material submitted to the VTH laboratory for diagnostic purposes or collected for research purposes in collaboration with local veterinarians. Stray cats were admitted to undergo neutering programs for the demographic control of stray population. Samples were stored at 4 °C until arrival at the laboratory (within 24 h from collection). The study was approved by the Institutional Animal Care and Use Committee and the Institutional Ethical Committee (approval no. OPBA_40_2020). Furthermore, residual fecal samples from cats collected for diagnostic purposes at the VTH with the informed consent of the owners were used for this study without any additional formal request for authorization, according to the decision of the Ethical Committee of the University of Milan (EC decision 29 October 2012, renewed with protocol no. 02-2016).

4.2. Identification of ESBL/AmpC-Producing E. coli

Samples were screened for the presence of ESBL/AmpC-producing E. coli according to the standard protocol of the DTU National Food Institute, the reference laboratory for antimicrobial resistance in Europe, with minor modifications [49]. One gram of each fecal sample was pre-enriched in buffered peptone water (BPW) and incubated at 37 ± 1 °C for 18–22 h. For ESBL/AmpC-producing Enterobacteriaceae, a loopful of BPW was inoculated on MacConkey agar supplemented with 1 mg/L cefotaxime and incubated overnight at 37 ± 1 °C. Up to four bacterial colonies from positive growths of each sample were submitted to species identification regardless of colony morphology (lactose +/−). Species identification was accomplished in duplicate via matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) (MBT Microflex ® LT/SH MALDI-TOF mass spectrometer, Bruker Daltonik, GmbH, Bremen, Germany) using the direct transfer method [50]. Following species identification, all four colonies/samples recovered from MacConkey agar supplemented with 1 mg/L cefotaxime were stored in brain heart infusion (BHI) broth with 15% glycerol at −80 °C.
One confirmed E. coli isolate for each sample was further thawed and used for ESBL and AmpC phenotypes confirmation using a combination disk test (CDT) and AmpC detection set D69C (MAST Group Ltd., Bootle, UK), respectively, according to EUCAST guidelines [33]. Briefly, a pure fresh culture of the tested isolate was suspended in physiological saline to obtain a 0.5 McFarland standard density equivalent suspension. The suspension was spread uniformly across the surface of a Mueller Hinton agar plate. Disks used for ESBL phenotyping contained ceftazidime or cefotaxime (30 μg) with and without clavulanic acid (10 μg). Disks used for AmpC phenotyping were disks A, B, and C. Disks were placed on the inoculated medium and incubated at 35 ± 1 °C for 18 ± 2 h. Results were interpreted following the manufacture’s instructions. For ESBL phenotyping, the isolate was considered positive in the presence of a ≥5 mm increase in the inhibition zone diameter in the disk containing ceftazidime or cefotaxime and clavulanic acid compared to the one without clavulanic acid. For AmpC phenotyping, the isolate was considered positive if the difference in the inhibition zone diameter between disks C and A and between disks C and B was ≥5 mm.
Isolates were also subjected to PCR analysis and the evaluation of minimum inhibitory concentrations (MICs).

4.3. PCR Analysis for Resistance Genes and E. coli Phylogroup

Single thawed E. coli colonies recovered from MacConkey agar supplemented with 1 mg/L cefotaxime were resuspended in 100 µL of sterile distilled water, and DNA was extracted by the boiling method at 95 °C for 10 min and subjected to PCR analysis to determine ESBL/AmpC genes and E. coli phylogroup.
For the detection of ESBL-producing E. coli, a multiplex PCR targeting blaCTX-M, blaTEM, and blaSHV was performed on the DNA of all isolates, as previously reported [51]. Isolates positive for blaCTX-M genes were further analyzed with a specific PCR for the blaCTX-M-1 group, blaCTX-M-2 group, blaCTX-M-8 group, blaCTX-M-9 group, and blaCTX-M-25 group, as previously reported [52,53,54,55,56]. Since the ESBL phenotype could be masked by the presence of AmpC β-lactamases and/or carbapenemases, cats were considered positive for the presence of ESBL-producing E. coli in the presence of at least one ESBL-encoding gene, regardless of phenotypic confirmation, as previously reported [33,57].
The DNA of E. coli isolates showing the AmpC phenotype were subjected to PCR for the detection of major plasmid-mediated ampC β-lactamase (pampC)-encoding genes and chromosomal-mediated ampC β-lactamases (campC). The presence of pampC was determined using a multiplex PCR, as previously reported [58]. The presence of campC hyperproduction was investigated by the amplification of a 271 bp fragment of the promoter/attenuator region, as previously reported [59]. Amplicons were purified and Sanger sequenced by a commercial sequencing facility (Microsynth Seqlab, Göttingen, Germany). The sequences were aligned against the promoter/attenuator region of the campC gene of E. coli strain ATCC 25922 using Clustal X in BioEdit software v.7.0. Strains were labeled campC hyperproducers when promoter mutations were found, according to previous reports [20,60]. E. coli isolates showing the AmpC phenotype supported by the overexpression of campC or by the presence of pampC genes were considered as AmpC-producing E. coli.
The E. coli phylogenetic group was determined following previously published protocols [44].
The different PCR assays were performed using positive control strains, and a blank control (DNAse-free water sample) was also included in all the PCR reactions. The primers used in this study are shown in Table S3.

4.4. Antimicrobial Susceptibility Testing

MICs were determined on single ESBL- and/or AmpC-producing E. coli isolates by the broth microdilution method using a commercially available plate (COMPGN1F Sensititre plates, Thermo Fisher Scientific®, Waltham, MA, USA). The list of antimicrobials used, the related cut-off values, and the MIC (µg/mL) range are reported in Table S4: E. coli ATCC 25922 was used as the control strain for susceptibility testing. Results were defined manually using a Sensititre Manual Viewbox (Sensititre™, Thermo Fisher Scientific®, Waltham, MA, USA). The MIC results were interpreted according to Clinical and Laboratory Standard Institute breakpoints CLSI VET01S ED6:2023 [61] following the manufacturer’s instructions. Multidrug resistance (MDR) was defined as non-susceptibility to at least one antimicrobial agent in three or more antimicrobial categories, as previously reported [62].

4.5. Data Analysis

For each animal, information regarding sex; age; type of ownership; and clinical history (clinical status, history of hospitalization, and previous antibiotic treatment performed within three months) were collected, as well as the results of biochemical analyses on blood and serum samples, if performed as part of the VTH diagnostic procedures. The stray cats belonged to 11 cat colonies from Lodi province, and the geographic location of the feline colony of origin was recorded. For the age variable, two categories were considered: <2 years old and ≥2 years old, as previously reported for the identification of age as a risk factor for infectious diseases [63]. The clinical status variable was classified into two categories: healthy and unhealthy, according to the presence/absence of a clinical manifestation and the results of routine laboratory analyses, if available. The clinical status of unhealthy cats was further classified, according to the main clinical presentation on admission, into gastrointestinal, respiratory, urogenital, dermatological, neurologic, traumatic, or systemic. Previous antibiotic treatment was further classified according to the antibiotic class used to treat the cats.
Pearson’s chi-square test and Fisher’s exact probability test were used to evaluate the differences between the proportions of ESBL/AmpC-producing E. coli-positive cats and sex, age, type of ownership, clinical status, hospitalization, and previous antibiotic therapy. Statistical comparisons were carried out using Epitools (https://epitools.ausvet.com.au/ (accessed on: 16 February 2023)), taking p < 0.05 as significant.

5. Conclusions

In this study, the fecal carriage of ESBL/AmpC-producing E. coli was detected in both owned and stray cats from Italy, with an estimated positivity of 6.2%, confirming that cats previously treated with antibiotics are at a higher risk of AMR carriage. These results highlight the need for antimicrobial stewardship in veterinary medicine and an AMR surveillance program focusing on companion animals, including stray cats.

Supplementary Materials

The following supporting information can be downloaded at: https://www.mdpi.com/article/10.3390/antibiotics12081249/s1, Table S1: Characteristics of unhealthy cats compared to healthy cats analyzed in this study, Table S2: Results of the chromosomal ampC promoter/attenuator region analysis of the AmpC-producing E. coli detected in this study, Table S3: Primer sequences used in this study, Table S4: List of antimicrobials used, related cut-off values, and the MIC (mg/L) range used in this study.

Author Contributions

Conceptualization, G.R. and S.L.; methodology, G.R.; validation, G.R. and S.L.; formal analysis, G.R., A.G. (Alessia Giordano), S.P. and S.L.; investigation, G.R., A.F. and M.P.; resources, A.S., P.S. and A.G. (Alessia Gazzonis); data curation, G.R., A.F. and D.M.; writing—original draft preparation, G.R.; writing—review and editing, A.G. (Alessia Giordano), S.P., P.D., C.L. and S.L.; visualization, G.R. and S.L.; supervision, S.L.; project administration, S.L.; funding acquisition, A.G. (Alessia Giordano), S.P., C.L., P.D. and S.L. All authors have read and agreed to the published version of the manuscript.

Funding

This work was funded by the University of Milan (Funding LINEA2_SLAUZ_2019_AA, Piano di Sostegno alla Ricerca UNIMI 2019: Zoonotic pathogens and antimicrobial resistance in pets: a model study from the One health—One welfare perspective, principal investigator Stefania Lauzi).

Institutional Review Board Statement

This study was approved by the Institutional Animal Care and Use Committee and the Institutional Ethical Committee (approval no. OPBA_40_2020). Furthermore, samples from owned cats were collected according to the diagnostic procedures and the Ethical Committee decision of the University of Milan. Residual aliquots of samples or tissues collected for diagnostic purposes at the VTH with the informed consent of the owners can be used for research purposes without any additional formal request of authorization (EC decision 29 October 2012, renewed with the protocol no. 02-2016).

Informed Consent Statement

Informed consent was obtained from the owners of the animals involved in the study.

Data Availability Statement

The data that supported the findings of this study are available from the corresponding author upon reasonable request.

Acknowledgments

The authors are grateful to all the veterinarians who contributed by helping in sample collection.

Conflicts of Interest

The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript; or in the decision to publish the results.

References

  1. World Health Organization. Antibiotic Resistance. Available online: https://www.who.int/news-room/fact-sheets/detail/antibiotic-resistance (accessed on 3 November 2022).
  2. Pomba, C.; Rantala, M.; Greko, C.; Baptiste, K.E.; Catry, B.; van Duijkeren, E.; Mateus, A.; Moreno, M.A.; Pyörälä, S.; Ružauskas, M.; et al. Public health risk of antimicrobial resistance transfer from companion animals. J. Antimicrob. Chemother. 2017, 72, 957–968. [Google Scholar] [CrossRef] [PubMed]
  3. Carvalho, I.; Safia Chenouf, N.; Cunha, R.; Martins, C.; Pimenta, P.; Pereira, A.R.; Martínez-Álvarez, S.; Ramos, S.; Silva, V.; Igrejas, G.; et al. Antimicrobial Resistance Genes and Diversity of Clones among ESBL-and Acquired AmpC-Producing Escherichia coli Isolated from Fecal Samples of Healthy and Sick Cats in Portugal. Antibiotics 2021, 10, 262. [Google Scholar] [CrossRef] [PubMed]
  4. Rubin, J.E.; Pitout, J.D. Extended-spectrum β-lactamase, carbapenemase and AmpC producing Enterobacteriaceae in companion animals. Vet. Microbiol. 2014, 170, 10–18. [Google Scholar] [CrossRef]
  5. World Health Organization. WHO Integrated Global Surveillance on ESBL-Producing E. coli Using a “One Health” Approach: Implementation and Opportunities; World Health Organization: Geneva, Switzerland, 2021. [Google Scholar]
  6. Commission Implementing Decision (EU) 2020/1729 of 17 November 2020 on the Monitoring and Reporting of Antimicrobial Resistance in Zoonotic and Commensal Bacteria and Repealing Implementing Decision 2013/652/EU. 2020. Available online: http://data.europa.eu/eli/dec_impl/2020/1729/oj (accessed on 7 November 2022).
  7. Guardabassi, L.; Schwarz, S.; Lloyd, D.H. Pet animals as reservoirs of antimicrobial-resistant bacteria. J. Antimicrob. Chemother. 2004, 54, 321–332. [Google Scholar] [CrossRef]
  8. Javed, M.U.; Ijaz, M.; Fatima, Z.; Anjum, A.A.; Aqib, A.I.; Ali, M.M.; Rehman, A.; Ahmed, A.; Ghaffar, A. Frequency and antimicrobial susceptibility of methicillin and vancomycin-resistant Staphylococcus aureus from bovine milk. Pak. Vet. J. 2021, 41, 463–468. [Google Scholar] [CrossRef]
  9. Ma, J.; Zhou, W.; Wu, J.; Liu, X.; Lin, J.; Ji, X.; Lin, H.; Wang, J.; Jiang, H.; Zhou, Q.; et al. Large-Scale Studies on Antimicrobial Resistance and Molecular Characterization of Escherichia coli from Food Animals in Developed Areas of Eastern China. Microbiol. Spectr. 2022, 10, e0201522. [Google Scholar] [CrossRef] [PubMed]
  10. Overgaauw, P.; Vinke, C.M.; Hagen, M.; Lipman, L. A One Health Perspective on the Human-Companion Animal Relationship with Emphasis on Zoonotic Aspects. Int. J. Environ. Res. Public Health 2020, 17, 3789. [Google Scholar] [CrossRef]
  11. Salgado-Caxito, M.; Benavides, J.A.; Adell, A.D.; Paes, A.C.; Moreno-Switt, A.I. Global prevalence and molecular characterization of extended-spectrum β-lactamase producing-Escherichia coli in dogs and cats-A scoping review and meta-analysis. One Health 2021, 12, 100236. [Google Scholar] [CrossRef]
  12. Ewers, C.; Bethe, A.; Semmler, T.; Guenther, S.; Wieler, L.H. Extended-spectrum β-lactamase-producing and AmpC-producing Escherichia coli from livestock and companion animals, and their putative impact on public health: A global perspective. Clin. Microbiol. Infect. 2012, 18, 646–655. [Google Scholar] [CrossRef] [Green Version]
  13. Formenti, N.; Grassi, A.; Parisio, G.; Romeo, C.; Guarneri, F.; Birbes, L.; Pitozzi, A.; Scali, F.; Maisano, A.M.; Boniotti, M.B.; et al. Extended-Spectrum-β-Lactamase-and AmpC-Producing Escherichia coli in Domestic Dogs: Spread, Characterisation and Associated Risk Factors. Antibiotics 2021, 10, 1251. [Google Scholar] [CrossRef]
  14. Melo, L.C.; Oresco, C.; Leigue, L.; Netto, H.M.; Melville, P.A.; Benites, N.R.; Saras, E.; Haenni, M.; Lincopan, N.; Madec, J.Y. Prevalence and molecular features of ESBL/pAmpC-producing Enterobacteriaceae in healthy and diseased companion animals in Brazil. Vet. Microbiol. 2018, 221, 59–66. [Google Scholar] [CrossRef] [PubMed]
  15. European Pet Food Federation. Available online: https://europeanpetfood.org/about/statistics/ (accessed on 30 November 2022).
  16. Zogg, A.L.; Simmen, S.; Zurfluh, K.; Stephan, R.; Schmitt, S.N.; Nüesch-Inderbinen, M. High Prevalence of Extended-Spectrum β-Lactamase Producing Enterobacteriaceae Among Clinical Isolates From Cats and Dogs Admitted to a Veterinary Hospital in Switzerland. Front. Vet. Sci. 2018, 5, 62. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  17. Piccolo, F.L.; Belas, A.; Foti, M.; Fisichella, V.; Marques, C.; Pomba, C. Detection of multidrug resistance and extended-spectrum/plasmid-mediated AmpC beta-lactamase genes in Enterobacteriaceae isolates from diseased cats in Italy. J. Feline Med. Surg. 2020, 22, 613–622. [Google Scholar] [CrossRef] [PubMed]
  18. Gargano, V.; Gambino, D.; Orefice, T.; Cirincione, R.; Castelli, G.; Bruno, F.; Interrante, P.; Pizzo, M.; Spada, E.; Proverbio, D.; et al. Can Stray Cats Be Reservoirs of Antimicrobial Resistance? Vet. Sci. 2022, 9, 631. [Google Scholar] [CrossRef] [PubMed]
  19. Dazio, V.; Nigg, A.; Schmidt, J.S.; Brilhante, M.; Mauri, N.; Kuster, S.P.; Brawand, S.G.; Schüpbach-Regula, G.; Willi, B.; Endimiani, A.; et al. Acquisition and carriage of multidrug-resistant organisms in dogs and cats presented to small animal practices and clinics in Switzerland. J. Vet. Intern. Med. 2021, 35, 970–979. [Google Scholar] [CrossRef]
  20. Coolen, J.P.M.; den Drijver, E.P.M.; Verweij, J.J.; Schildkraut, J.A.; Neveling, K.; Melchers, W.J.G.; Kolwijck, E.; Wertheim, H.F.L.; Kluytmans, J.A.J.W.; Huynen, M.A. Genome-wide analysis in Escherichia coli unravels a high level of genetic homoplasy associated with cefotaxime resistance. Microb. Genom. 2021, 7, 000556. [Google Scholar] [CrossRef]
  21. Silva, J.; Menezes, J.; Marques, C.; Pomba, C.F. Companion Animals-An Overlooked and Misdiagnosed Reservoir of Carbapenem Resistance. Antibiotics 2022, 11, 533. [Google Scholar] [CrossRef]
  22. Weese, J.S.; O’Brien, T.; Bateman, S. Fecal shedding of extended-spectrum beta-lactamase-producing Enterobacterales in cats admitted to an animal shelter. J. Feline Med. Surg. 2022, 24, 1301–1304. [Google Scholar] [CrossRef]
  23. Hordijk, J.; Schoormans, A.; Kwakernaak, M.; Duim, B.; Broens, E.; Dierikx, C.; Mevius, D.; Wagenaar, J.A. High prevalence of fecal carriage of extended spectrum β-lactamase/AmpC-producing Enterobacteriaceae in cats and dogs. Front. Microbiol. 2013, 4, 242. [Google Scholar] [CrossRef] [Green Version]
  24. Sfaciotte, R.; Parussolo, L.; Melo, F.D.; Wildemann, P.; Bordignon, G.; Israel, N.D.; Leitzke, M.; Wosiacki, S.R.; Salbego, F.Z.; da Costa, U.M.; et al. Identification and Characterization of Multidrug-Resistant Extended-Spectrum Beta-Lactamase-Producing Bacteria from Healthy and Diseased Dogs and Cats Admitted to a Veterinary Hospital in Brazil. Microb. Drug Resist. 2021, 27, 855–864. [Google Scholar] [CrossRef]
  25. Abbas, R.; Nawaz, Z.; Siddique, A.B.; Aslam, R.; Rafique, A.; Zahoor, M.A.; Qamar, M.U.; Ahmad, M.Z.; Jalees, M.M.; Qasim, M.; et al. Molecular detection of biofilm production among multidrug resistant isolates of Pseudomonas aeruginosa from meat samples. Pak. Vet. J. 2022, 42, 505–510. [Google Scholar]
  26. Liebana, E.; Carattoli, A.; Coque, T.M.; Hasman, H.; Magiorakos, A.P.; Mevius, D.; Peixe, L.; Poirel, L.; Schuepbach-Regula, G.; Torneke, K.; et al. Public health risks of enterobacterial isolates producing extended-spectrum β-lactamases or AmpC β-lactamases in food and food-producing animals: An EU perspective of epidemiology, analytical methods, risk factors, and control options. Clin. Infect. Dis. 2013, 56, 1030–1037. [Google Scholar] [CrossRef] [Green Version]
  27. Bush, K.; Jacoby, G.A. Updated functional classification of beta-lactamases. Agents Chemother. 2010, 54, 969–976. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  28. Peirano, G.; Pitout, J.D.D. Extended-Spectrum β-Lactamase-Producing Enterobacteriaceae: Update on Molecular Epidemiology and Treatment Options. Drugs 2019, 79, 1529–1541. [Google Scholar] [CrossRef] [PubMed]
  29. Bogaerts, P.; Huang, T.D.; Bouchahrouf, W.; Bauraing, C.; Berhin, C.; El Garch, F.; Glupczynski, Y.; ComPath Study Group. Characterization of ESBL-and AmpC-Producing Enterobacteriaceae from Diseased Companion Animals in Europe. Microb. Drug. Resist. 2015, 21, 643–650. [Google Scholar] [CrossRef] [PubMed]
  30. Rocha-Gracia, R.C.; Cortés-Cortés, G.; Lozano-Zarain, P.; Bello, F.; Martínez-Laguna, Y.; Torres, C. Faecal Escherichia coli isolates from healthy dogs harbour CTX-M-15 and CMY-2 β-lactamases. Vet. J. 2015, 203, 315–319. [Google Scholar] [CrossRef] [PubMed]
  31. Coolen, J.P.M.; den Drijver, E.P.M.; Kluytmans, J.A.J.W.; Verweij, J.J.; Lamberts, B.A.; Soer, J.A.C.J.; Verhulst, C.; Wertheim, H.F.L.; Kolwijck, E. Development of an algorithm to discriminate between plasmid- and chromosomal-mediated AmpC β-lactamase production in Escherichia coli by elaborate phenotypic and genotypic characterization. J. Antimicrob. Chemother. 2019, 74, 3481–3488. [Google Scholar] [CrossRef]
  32. Carattoli, A.; Lovari, S.; Franco, A.; Cordaro, G.; Di Matteo, P.; Battisti, A. Extended-spectrum beta-lactamases in Escherichia coli isolated from dogs and cats in Rome, Italy, from 2001 to 2003. Antimicrob. Agents Chemother. 2005, 49, 833–835. [Google Scholar] [CrossRef] [Green Version]
  33. EUCAST Guidelines for Detection of Resistance Mechanisms and Specific Resistances of Clinical and/or Epidemiological Importance (V 2.0 July 2017). Available online: https://www.eucast.org/fileadmin/src/media/PDFs/EUCAST_files/Resistance_mechanisms/EUCAST_detection_of_resistance_mechanisms_170711.pdf (accessed on 7 November 2022).
  34. Miller, S.; Humphries, R.M. Clinical laboratory detection of carbapenem-resistant and carbapenemase-producing Enterobacteriaceae. Expert. Rev. Anti. Infect. Ther. 2016, 14, 705–717. [Google Scholar] [CrossRef]
  35. Vlek, A.L.; Frentz, D.; Haenen, A.; Bootsma, H.J.; Notermans, D.W.; Frakking, F.N.; de Greeff, S.C.; Leenstra, T.; ISIS-AR study group. Detection and epidemiology of carbapenemase producing Enterobacteriaceae in the Netherlands in 2013-2014. Eur. J. Clin. Microbiol. Infect. Dis. 2016, 35, 1089–1096. [Google Scholar] [CrossRef]
  36. Tamma, P.D.; Huang, Y.; Opene, B.N.; Simner, P.J. Determining the Optimal Carbapenem MIC That Distinguishes Carbapenemase-Producing and Non-Carbapenemase-Producing Carbapenem-Resistant Enterobacteriaceae. Antimicrob. Agents Chemother. 2016, 60, 6425–6429. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  37. Ong, D.S.Y.; Altorf-van der Kuil, W.; Vlek, A.L.M.; Schouls, L.M.; Schoffelen, A.F. Routinely available antimicrobial susceptibility information can be used to increase the efficiency of screening for carbapenemase-producing Enterobacteriaceae. J. Med. Microbiol. 2020, 69, 1235–1239. [Google Scholar] [CrossRef] [PubMed]
  38. Hong, J.S.; Song, W.; Jeong, S.H. Molecular Characteristics of NDM-5-Producing Escherichia coli from a Cat and a Dog in South Korea. Microb. Drug Resist. 2020, 26, 1005–1008. [Google Scholar] [CrossRef] [PubMed]
  39. Cole, S.D.; Peak, L.; Tyson, G.H.; Reimschuessel, R.; Ceric, O.; Rankin, S.C. New Delhi Metallo-β-Lactamase-5–Producing Escherichia coli in Companion Animals, United States. Emerg. Infect. Dis. 2020, 26, 381–383. [Google Scholar] [CrossRef] [Green Version]
  40. Bortolami, A.; Zendri, F.; Maciuca, E.I.; Wattret, A.; Ellis, C.; Schmidt, V.; Pinchbeck, G.; Timofte, D. Diversity, Virulence, and Clinical Significance of Extended-Spectrum β-Lactamase- and pAmpC-Producing Escherichia coli From Companion Animals. Front. Microbiol. 2019, 10, 1260. [Google Scholar] [CrossRef] [Green Version]
  41. Giufrè, M.; Mazzolini, E.; Cerquetti, M.; Brusaferro, S.; CCM2015 One-Health ESBL-producing Escherichia coli Study Group. Extended-spectrum β-lactamase-producing Escherichia coli from extraintestinal infections in humans and from food-producing animals in Italy: A ‘One Health’ study. Int. J. Antimicrob. Agents 2021, 58, 106433. [Google Scholar] [CrossRef]
  42. Zhou, Y.; Ji, X.; Liang, B.; Jiang, B.; Li, Y.; Yuan, T.; Zhu, L.; Liu, J.; Guo, X.; Sun, Y. Antimicrobial Resistance and Prevalence of Extended Spectrum β-Lactamase-Producing Escherichia coli from Dogs and Cats in Northeastern China from 2012 to 2021. Antibiotics 2022, 11, 1506. [Google Scholar] [CrossRef]
  43. Jakobsen, L.; Garneau, P.; Kurbasic, A.; Bruant, G.; Stegger, M.; Harel, J.; Jensen, K.S.; Brousseau, R.; Hammerum, A.M.; Frimodt-Møller, N. Microarray-based detection of extended virulence and antimicrobial resistance gene profiles in phylogroup B2 Escherichia coli of human, meat and animal origin. J. Med. Microbiol. 2011, 60, 1502–1511. [Google Scholar] [CrossRef]
  44. Clermont, O.; Christenson, J.K.; Denamur, E.; Gordon, D.M. The Clermont Escherichia coli phylo-typing method revisited: Improvement of specificity and detection of new phylo-groups. Environ. Microbiol. Rep. 2013, 5, 58–65. [Google Scholar] [CrossRef]
  45. Walther, B.; Tedin, K.; Lübke-Becker, A. Multidrug-resistant opportunistic pathogens challenging veterinary infection control. Vet. Microbiol. 2017, 200, 71–78. [Google Scholar] [CrossRef]
  46. Ortiz-Díez, G.; Mengíbar, R.L.; Turrientes, M.C.; Artigao, M.B.; Gallifa, R.L.; Tello, A.M.; Pérez, C.F.; Santiago, T.A. Prevalence, incidence and risk factors for acquisition and colonization of extended-spectrum beta-lactamase- and carbapenemase-producing Enterobacteriaceae from dogs attended at a veterinary hospital in Spain. Comp. Immunol. Microbiol. Infect. Dis. 2023, 92, 101922. [Google Scholar] [CrossRef]
  47. Shnaiderman-Torban, A.; Navon-Venezia, S.; Kelmer, E.; Cohen, A.; Paitan, Y.; Arielly, H.; Steinman, A. Extended-Spectrum β-Lactamase-Producing Enterobacterales Shedding by Dogs and Cats Hospitalized in an Emergency and Critical Care Department of a Veterinary Teaching Hospital. Antibiotics 2020, 9, 545. [Google Scholar] [CrossRef]
  48. Marchetti, L.; Buldain, D.; Gortari Castillo, L.; Buchamer, A.; Chirino-Trejo, M.; Mestorino, N. Pet and Stray Dogs as Reservoirs of Antimicrobial-Resistant Escherichia coli. Int. J. Microbiol. 2021, 2021, 6664557. [Google Scholar] [CrossRef]
  49. Isolation of ESBL-, AmpC- and Carbapenemase-Producing E. coli from Caecal Samples (V 7 2019). Available online: https://www.eurl-ar.eu/CustomerData/Files/Folders/21-protocols/530_esbl-ampc-cpeprotocol-version-caecal-v7-09-12-19.pdf (accessed on 6 November 2022).
  50. Singhal, N.; Kumar, M.; Kanaujia, P.K.; Virdi, J.S. MALDI-TOF mass spectrometry: An emerging technology for microbial identification and diagnosis. Front. Microbiol. 2015, 6, 791. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  51. Monstein, H.J.; Ostholm-Balkhed, A.; Nilsson, M.V.; Nilsson, M.; Dornbusch, K.; Nilsson, L.E. Multiplex PCR amplification assay for the detection of blaSHV, blaTEM and blaCTX-M genes in Enterobacteriaceae. APMIS 2007, 115, 1400–1408. [Google Scholar] [CrossRef] [PubMed]
  52. Saladin, M.; Cao, V.T.; Lambert, T.; Donay, J.L.; Herrmann, J.L.; Ould-Hocine, Z.; Verdet, C.; Delisle, F.; Philippon, A.; Arlet, G. Diversity of CTX-M beta-lactamases and their promoter regions from Enterobacteriaceae isolated in three Parisian hospitals. FEMS. Microbiol. Lett. 2002, 209, 161–168. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  53. Bertrand, S.; Weill, F.X.; Cloeckaert, A.; Vrints, M.; Mairiaux, E.; Praud, K.; Dierick, K.; Wildemauve, C.; Godard, C.; Butaye, P.; et al. Clonal emergence of extended-spectrum beta-lactamase (CTX-M-2)-producing Salmonella enterica serovar Virchow isolates with reduced susceptibilities to ciprofloxacin among poultry and humans in Belgium and France (2000 to 2003). J. Clin. Microbiol. 2006, 44, 2897–2903. [Google Scholar] [CrossRef] [Green Version]
  54. Jouini, A.; Vinué, L.; Slama, K.B.; Sáenz, Y.; Klibi, N.; Hammami, S.; Boudabous, A.; Torres, C. Characterization of CTX-M and SHV extended-spectrum beta-lactamases and associated resistance genes in Escherichia coli strains of food samples in Tunisia. J. Antimicrob. Chemother. 2007, 60, 1137–1141. [Google Scholar] [CrossRef] [Green Version]
  55. Jiang, X.; Ni, Y.; Jiang, Y.; Yuan, F.; Han, L.; Li, M.; Liu, H.; Yang, L.; Lu, Y. Outbreak of infection caused by Enterobacter cloacae producing the novel VEB-3 beta-lactamase in China. J. Clin. Microbiol. 2005, 43, 826–831. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  56. Chmelnitsky, I.; Carmeli, Y.; Leavitt, A.; Schwaber, M.J.; Navon-Venezia, S. CTX-M-2 and a new CTX-M-39 enzyme are the major extended-spectrum beta-lactamases in multiple Escherichia coli clones isolated in Tel Aviv, Israel. Agents Chemother. 2005, 49, 4745–4750. [Google Scholar] [CrossRef] [Green Version]
  57. Garrec, H.; Drieux-Rouzet, L.; Golmard, J.L.; Jarlier, V.; Robert, J. Comparison of nine phenotypic methods for detection of extended-spectrum beta-lactamase production by Enterobacteriaceae. J. Clin. Microbiol. 2011, 49, 1048–1057. [Google Scholar] [CrossRef] [Green Version]
  58. Dallenne, C.; Da Costa, A.; Decré, D.; Favier, C.; Arlet, G. Development of a set of multiplex PCR assays for the detection of genes encoding important beta-lactamases in Enterobacteriaceae. J. Antimicrob. Chemother. 2010, 65, 490–495. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  59. Corvec, S.; Prodhomme, A.; Giraudeau, C.; Dauvergne, S.; Reynaud, A.; Caroff, N. Most Escherichia coli strains overproducing chromosomal AmpC beta-lactamase belong to phylogenetic group A. J. Antimicrob. Chemother. 2007, 60, 872–876. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  60. Peter-Getzlaff, S.; Polsfuss, S.; Poledica, M.; Hombach, M.; Giger, J.; Böttger, E.C.; Zbinden, R.; Bloemberg, G.V. Detection of AmpC beta-lactamase in Escherichia coli: Comparison of three phenotypic confirmation assays and genetic analysis. J. Clin. Microbiol. 2011, 49, 2924–2932. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  61. Clinical and Laboratory Standards Institute (CLSI). Performance Standards for Antimicrobial Disk and Dilution Susceptibility Tests for Bacteria Isolated from Animals, 6th ed.; CLSI Supplement VET01S; Clinical and Laboratory Standards Institute: Wayne, PA, USA, 2023; ISBN 978-1-68440-167-3. [Google Scholar]
  62. Magiorakos, A.P.; Srinivasan, A.; Carey, R.B.; Carmeli, Y.; Falagas, M.E.; Giske, C.G.; Harbarth, S.; Hindler, J.F.; Kahlmeter, G.; Olsson-Liljequist, B.; et al. Multidrug-resistant, extensively drug-resistant and pandrug-resistant bacteria: An international expert proposal for interim standard definitions for acquired resistance. Clin. Microbiol. Infect. 2012, 18, 268–281. [Google Scholar] [CrossRef] [Green Version]
  63. Anpuanandam, K.; Selvarajah, G.T.; Choy, M.; Ng, S.W.; Kumar, K.; Ali, R.M.; Rajendran, S.K.; Ho, K.L.; Tan, W.S. Molecular detection and characterization of Domestic Cat Hepadnavirus (DCH) from blood and liver tissues of cats in Malaysia. BMC Vet. Res. 2021, 17, 9. [Google Scholar] [CrossRef]
Table
Table 1. Characteristics of cats analyzed in this study.
Table 1. Characteristics of cats analyzed in this study.
Population Characteristics No. (%)No. ESBL/AmpC-Positive (%)p Value a
SexMale54 (55.7)5 (9.3)0.22
Female43 (44.3)1 (2.3)
Age b<2 years55 (57.9)3 (5.5)1
≥2 years40 (42.1)3 (7.5)
Type of ownershipStray50 (51.5)2 (4.0)0.43
Owned47 (48.5)4 (8.5)
Clinical statusHealthy70 (72.2)2 (2.9)0.049
Unhealthy27 (27.8)4 (14.8)
Clinical syndrome at admissionGastrointestinal12 (44.4)2 (16.7)nd
Respiratory8 (29.6)1 (12.5)
Urogenital2 (7.4)0 (0)
Systemic2(7.4)1 (50)
Dermatological1 (3.7)0 (0)
Neurologic1 (3.7)0 (0)
Traumatic1 (3.7)0 (0)
HospitalizationYes10 (10.3)2 (20.0)0.11
No87 (89.7)4 (4.6)
Previous antibiotic therapyYes25 (25.8)4 (16)0.037
No72 (74.2)2 (2.8)
Antibiotic class used in treated cats cFluoroquinolones12 (48.0)0 (0)nd
β-Lactams and β-Lactamase inhibitors10 (40.0)1 (10.0)
Cephalosporins7 (28.0)2 (28.6)
Macrolide-nitroimidazole2 (8.0)1 (50.0)
Tetracyclines2 (8.0)0 (0)
a Numbers in bold indicate p < 0.05. b Age was unknown for two cats. c Cats treated with single and combination therapy were included. nd = not determined. Statistical analysis was not performed due to the low numbers of cats included in the different groups.
Table
Table 2. Phenotypic and genetic characterization of ESBL/AmpC-producing E. coli isolates.
Table 2. Phenotypic and genetic characterization of ESBL/AmpC-producing E. coli isolates.
Isolate IDAnimal CharacteristicsPhenotypeGenetic Determinants of ResistancePhylogenetic GroupResistance Pattern
SexAgeOwnershipClinical StatusHospitalizationPrevious Antibiotic Therapy (Antibiotic)
46/1Female6 monthsStrayHealthyNoNoESBL/AmpCblaCTX-M-1 group, campC hyperproducerB2AMP, FAZ, FOV, POD, LEX, AUG2, DOX, TET
51/1Male3 yearsStrayHealthyNoNoESBLblaCTX-M-1 group, blaTEMAAMP, FAZ, FOV, POD, LEX, DOX, TET
77/1Male13 yearsOwnedUnhealthyNoYes (cephalosporin)ESBLblaCTX-M-1 group, blaTEMB2AMP, FAZ, FOV, POD, LEX, DOX, TET, SXT
137/1Male1 yearOwnedUnhealthyYesYes (cephalosporin)ESBLblaCTX-M-1 group, blaTEM, blaSHVB2AMP, FAZ, FOV, POD, LEX, DOX, TET, SXT
161/1Male7 yearsOwnedUnhealthyYesYes (amoxicillin + clavulanic acid)ESBLblaCTX-M-1 group, blaTEMB2AMP, FAZ, FOV, POD, LEX, AUG2, DOX, CHL, GEN, TET, SXT
195/1Male10 monthsOwnedUnhealthyNoYes (metronidazole and spiramycin)NegativeblaCTX-M-1 group, blaTEMFAMP, FAZ, FOV, POD, LEX, DOX, TAZ, AUG2, P/T4, IMI, GEN, ENRO, MAR, ORB, PRA
AMP: ampicillin; AUG2: amoxicillin/clavulanic acid 2:1 (value refers to amoxicillin concentration); CHL: chloramphenicol; DOX: doxycycline; ENRO: enrofloxacin; FAZ: cefazolin; FOV: cefovecin; GEN: gentamicin; IMI: imipenem; LEX: cephalexin; MAR: marbofloxacin; ORB: orbifloxacin; PRA: pradofloxacin; POD: cefpodoxime; P/T4: piperacillin/tazobactam constant 4; SXT: trimethoprim/sulfamethoxazole 1:19 (value refers to trimethoprim concentration); TAZ: ceftazidime; TET: tetracycline.
Table
Table 3. Distribution of MICs among ESBL/AmpC-producing E. coli isolates.
Table 3. Distribution of MICs among ESBL/AmpC-producing E. coli isolates.
Antimicrobial ClassAntimicrobial AgentNo. ResistantNo. of Isolates at the Indicated MIC µg/mL
0.120.250.51248163264128
β-Lactam (penicillins)AMP6/6 6
β-Lactam (cephalosporin I)FAZ6/6 6
LEX6/6 6
β-Lactam (cephalosporin III)FOV6/6 6
POD6/6 6
TAZ1/6 23 1
β-Lactams and β-lactamase inhibitorsAUG23/6 123
P/T41/6 41 1
AminoglycosidesAMI0/6 51
GEN2/6 21 1 2
PhenicolCHL1/6 131 1
FluoroquinolonesENRO1/632 1
MAR1/632 1
ORB1/6 32 1
PRA1/6 5 1
Tetracyclines TET5/6 1 5
DOX6/6 21 12
CarbapenemsIMI1/6 5 1
Folate pathway antagonistsSXT3/6 3 3
White fields denote the range of dilutions tested for each antimicrobial agent. Grey fields denote the range of dilutions not tested for each antimicrobial agent Vertical lines indicate CLSI VET01S ED6:2023 cut-off values (intermediate results were considered susceptible). AMP: ampicillin; FAZ: cefazolin; LEX: cephalexin; FOV: cefovecin; POD: cefpodoxime; TAZ: ceftazidime; AUG2: amoxicillin/clavulanic acid 2:1 (value refers to amoxicillin concentration); P/T4: piperacillin/tazobactam constant 4; AMI: amikacin; GEN: gentamicin; CHL: chloramphenicol; ENRO: enrofloxacin; MAR: marbofloxacin; ORB: orbifloxacin; PRA: pradofloxacin; TET: tetracycline; DOX: doxycycline; IMI: imipenem; SXT: trimethoprim/sulfamethoxazole 1:19 (value refers to trimethoprim concentration).
Disclaimer/Publisher’s Note: The statements, opinions and data contained in all publications are solely those of the individual author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to people or property resulting from any ideas, methods, instructions or products referred to in the content.

Share and Cite

MDPI and ACS Style

Ratti, G.; Facchin, A.; Stranieri, A.; Giordano, A.; Paltrinieri, S.; Scarpa, P.; Maragno, D.; Gazzonis, A.; Penati, M.; Luzzago, C.; et al. Fecal Carriage of Extended-Spectrum β-Lactamase-/AmpC-Producing Escherichia coli in Pet and Stray Cats. Antibiotics 2023, 12, 1249. https://doi.org/10.3390/antibiotics12081249

AMA Style

Ratti G, Facchin A, Stranieri A, Giordano A, Paltrinieri S, Scarpa P, Maragno D, Gazzonis A, Penati M, Luzzago C, et al. Fecal Carriage of Extended-Spectrum β-Lactamase-/AmpC-Producing Escherichia coli in Pet and Stray Cats. Antibiotics. 2023; 12(8):1249. https://doi.org/10.3390/antibiotics12081249

Chicago/Turabian Style

Ratti, Gabriele, Alessia Facchin, Angelica Stranieri, Alessia Giordano, Saverio Paltrinieri, Paola Scarpa, Deborah Maragno, Alessia Gazzonis, Martina Penati, Camilla Luzzago, and et al. 2023. "Fecal Carriage of Extended-Spectrum β-Lactamase-/AmpC-Producing Escherichia coli in Pet and Stray Cats" Antibiotics 12, no. 8: 1249. https://doi.org/10.3390/antibiotics12081249

Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details here.

Article Metrics

Back to TopTop