Investigation on biofilm composition and virulence traits of S. pseudintermedius isolated from infected and colonized dogs.

Staphylococcus pseudintermedius, which is part of the skin microbiome of dogs, causes a variety of opportunistic infections. These infections may become more difficult to treat due to the formation of biofilm. The capacity of S. pseudintermedius to form biofilm, as well as the associated genes, has not been elucidated. This study evaluated the production and composition of S. pseudintermedius biofilm. Samples were collected from both infected dogs and asymptomatic dogs. Isolates were identified using mass spectrometry and Multiplex-PCR. Biofilm production and composition were assessed using a quantitative microtiter plate assay. The presence of ica operon genes and sps genes was investigated using conventional PCR. The investigation of Agr type and virulence genes was conducted in silico on 24 sequenced samples. All strains could produce strong biofilms, with most of the isolates presenting a polysaccharide biofilm. 63.6% of the isolates carried the complete ica operon (ADBC). All samples showed the presence of the genes spsK, spsA, and spsL, while the distribution of other genes varied. Agr type III was the most prevalent (52.2%). All sequenced samples carried the cytotoxins hlb, luk-S, luk-F, as well as the exfoliative toxins siet and se_int. No isolate displayed other exfoliative toxins. Only LB1733 presented a set of different enterotoxins (sea, seb, sec_canine, seh, sek, sel, and seq). Our findings suggest that S. pseudintermedius is a strong producer of biofilm and carries virulence genes.

Biofilm formation by Staphylococcus pseudintermedius on titanium implants.

Osteomyelitis often involves Staphylococcus spp. as the isolated genus in domestic animal cases. Implant-related infections, frequently associated with biofilm-forming microorganisms like staphylococci species, necessitate careful material selection. This study assessed biofilm formation by Staphylococcus pseudintermedius on titanium nuts used in veterinary orthopaedic surgery. Biofilm quantification employed safranin staining and spectrophotometric measurement, while bacterial counts were determined in colony-forming units (CFU). Scanning Electron Microscopy (SEM) evaluated the biofilm morphology on the surface of titanium nuts. All samples had CFU counts. Absorbance values that evidence biofilm formation were observed in seven of the eight samples tested. SEM images revealed robust bacterial colonization, and significant extracellular polymeric substance production, and the negative control displayed surface irregularities on the nut. Whole genome sequencing revealed accessory Gene Regulator (agr) type III in six samples, agr IV and agr II in two each. Genes encoding hlb, luk-S, luk-F, siet, se_int, and the icaADCB operon were identified in all sequenced samples. Other exfoliative toxins were absent. Biofilm formation by S. pseudintermedius was detected in all samples, indicating the susceptibility of orthopaedic titanium alloys to adhesion and biofilm formation by veterinary species. The biofilm formation capacity raises concerns about potential post-surgical complications and associated costs.

Epidemiologic case investigation on the zoonotic transmission of Methicillin-resistant Staphylococcus pseudintermedius among dogs and their owners.

Dogs often carry methicillin-resistant Staphylococci asymptomatically. These bacteria are frequently linked to conditions such as canine pyoderma and otitis. Close interaction between dogs and humans can facilitate the exchange of resistant strains, particularly Methicillin-resistant Staphylococcus pseudintermedius (MRSP). This represents a public health issue, since these strains, in addition to occasionally causing infections in humans, can also serve as a source of resistance and virulence genes for strains of greater importance in human medicine, such as Staphylococcus aureus. Furthermore, MRSP strains are often multidrug resistant, which ends up compromising the treatment of infections. This study aimed to assess the potential transmission of Staphylococcus pseudintermedius among dogs and their owners. We examined a total of one hundred canine samples collected from cases of pyoderma and otitis to detect the presence of staphylococci. Simultaneously, we conducted evaluations on all dog owners. Staphylococci strains were identified using MALDI-TOF MS and PCR targeting the nuc gene. Methicillin resistance screening was also performed by detecting the mecA gene using PCR. Among the sampled dogs, 64 carried S. pseudintermedius. Nine were identified as MRSP. In six instances, dogs and their owners exhibited S. pseudintermedius. These samples underwent genome sequencing and were screened for antimicrobial resistance genes, SCCmec typing, MLST characterization, and Single Nucleotide Polymorphisms (SNP) analyses. The results of the phylogenetic analysis revealed that in three cases, dogs and owners had closely related isolates, suggesting interspecies transmission. Two of these cases involved MRSP and one MSSP. Moreover, in the two MRSP cases, the same SCCmec type (type V) was detected. Additionally, the sequence type was consistent across all three cases involving dogs and owners (MSSP ST2277, MRSP ST2282, and ST2286). These findings strongly indicate a transmission event. Since Staphylococcus pseudintermedius is primarily isolated from canine samples, it is plausible that dogs may have acted as a potential source. In the remaining three cases, despite identifying the same species in both samples, they had notable phylogenetic differences.

The Diversity and Zoonotic Potential of Staphylococcus pseudintermedius in Humans and Pet Dogs in Trinidad and Tobago.

Staphylococcus pseudintermedius is an opportunistic pathogen that is frequently isolated from canines. It is of escalating interest because of its increasing antimicrobial resistance and zoonotic potential. Although many published articles are available that describe isolates obtained from diseased dogs and humans, this study focused on isolates obtained from healthy dogs and their owners who presented at clinics for routine veterinary care and utilized whole genome sequencing-based analyses for strain comparisons. A total of 25 humans and 27 canines were sampled at multiple sites, yielding 47 and 45 isolates, respectively. Whole genome sequence analysis was performed. We detected mostly new sequence types (STs) and a high diversity. Strains carried few antimicrobial resistance genes and plasmids, albeit three MRSP strains were found that belonged to two internationally distributed STs. The virulence content did not provide insights toward a tendency to colonization of humans but supported that there may be differences in the surface proteins between carrier strains and those causing pyoderma. We identified 13 cases in which humans were infected with strains from the dog they owned.

Genomic Features of Antimicrobial Resistance in Staphylococcus pseudintermedius Isolated from Dogs with Pyoderma in Argentina and the United States: A Comparative Study.

Staphylococcus pseudintermedius is the most common opportunistic pathogen in dogs and methicillin resistance (MRSP) has been identified as an emerging problem in canine pyoderma. Here, we evaluated the antimicrobial resistance (AMR) features and phylogeny of S. pseudintermedius isolated from canine pyoderma cases in Argentina (n = 29) and the United States (n = 29). 62% of isolates showed multi-drug resistance. The AMR genes found: mecA, blaZ, ermB, dfrG, catA, tetM, aac(6')-aph(2″), in addition to tetK and lnuA (only found in U.S. isolates). Two point mutations were detected: grlA(S80I)-gyrA(S84L), and grlA(D84N)-gyrA(S84L) in one U.S. isolate. A mutation in rpoB (H481N) was found in two isolates from Argentina. SCCmec type III, SCCmec type V, ΨSCCmec57395 were identified in the Argentinian isolates; and SCCmec type III, SCCmec type IVg, SCCmec type V, and SCCmec type VII variant in the U.S. cohort. Sequence type (ST) ST71 belonging to a dominant clone was found in isolates from both countries, and ST45 only in Argentinian isolates. This is the first study to comparatively analyze the population structure of canine pyoderma-associated S. pseudintermedius isolates in Argentina and in the U.S. It is important to maintain surveillance on S. pseudintermedius populations to monitor AMR and gain further understanding of its evolution and dissemination.

Standardization of in vitro dual-species biofilms of Staphylococcus pseudintermedius and Malassezia pachydermatis: a strategy to establish an ex vivo biofilm model.

Ex vivo experiments have been performed aiming at mimicking in vivo environments. The main aim of this research was to standardize in vitro dual-species biofilm formation by Staphylococcus pseudintermedius and Malassezia pachydermatis as a strategy to establish an ex vivo biofilm model. Initially, the in vitro formation of biofilms in co-culture was established, using YPD medium, inoculum turbidity of 0.5 on the McFarland scale and maturation periods of 96 h for M. pachydermatis and 48 h for S. pseudintermedius. Subsequently, biofilms were formed on porcine skin using the same conditions, under which a greater number of cells/ml was observed in in vitro dual-species than in in vitro mono-species biofilms. Furthermore, ex vivo biofilm images demonstrated the formation of a highly structured biofilm with the presence of cocci and yeasts surrounded by the matrix. Thus, these conditions optimized the growth of both microorganisms within biofilms in vitro and ex vivo.

First report of Staphylococcus pseudintermedius ST71-SCCmec III and ST45-ΨSCCmec57395 from canine pyoderma in Argentina.

Staphylococcus pseudintermedius is an opportunistic pathogen commonly associated with skin infections in dogs. Twenty-three methicillin-resistant S. pseudintermedius (MRSP) isolated in Argentina from dogs with pyoderma were analyzed using whole genome sequencing (WGS) and classified into sequence types (ST) by multilocus sequence typing (MLST) and staphylococcal chromosome cassette mec (SCCmec) types.Based on the WGS analysis, MLST, and SCCmec type results, we report for the first time in Argentina two MRSP strains, one each, belonging to ST71-SCCmec III and ST45-ΨSCCmec57395 from dogs with pyoderma. We also identified seven isolates with ST339, which had been previously reported in only two isolates in Argentina. Additionally, we identified ten MRSP isolates harboring variants of the SCCmec V found in S. aureus, seven SCCmec V (5C2&5) with two ccrC1 recombinases, and three SCCmec V (5C2) with one ccrC1 recombinase.Our findings provide important insights into the evolution and geographic spread of these hypervirulent dominant clones that threaten the health of our companion animals and represent a significant risk for zoonotic infections.

Detection of the international lineage ST71 of methicillin-resistant Staphylococcus pseudintermedius in two cities in Rio de Janeiro State.

Staphylococcus pseudintermedius is the main coagulase-positive staphylococci associated with canine skin/soft tissue infections (SSTI), otitis externa, and surgical site infections. The international spread of an epidemic and multiresistant lineage of methicillin-resistant Staphylococcus pseudintermedius (MRSP), the so-called European clone-displaying sequence type (ST) 71-requires attention. The first isolation of an MRSP ST71 isolate in South America was reported in Rio de Janeiro city, in 2010; however, a limited number of canine isolates were analyzed. Thus, to have a better panel of the MRSP spread in this city, we were stimulated to continue this study and search for the presence of MRSP in 282 colonized or infected dogs in the city of Rio de Janeiro. Among the MRSP isolates collected (N = 17; 6.1%), the pulsed-field gel electrophoresis (PFGE) patterns were similar to those of European clone. All 17 isolates were classified as ST71 by multilocus sequence typing (MLST). In order to assess whether isolates of MRSP ST71 may have also spread to the Rio de Janeiro state countryside, we collected samples from 124 infected dogs in the city of Campos dos Goytacazes (232 km away from Rio de Janeiro city). Our data showed the presence of ST71 lineage in one isolate among three MRSP detected. S. pseudintermedius was isolated from 40.6% of the clinical samples (N = 165/406). A relatively high incidence of methicillin resistance, detected by a PCR-based method, was found in 12.1% of the S. pseudintermedius recovered from animals (N = 20/165). The resistance profile of these isolates was similar to that described for the international ST71 strains whose genomes are publicly available in the GenBank. The prospect of ST71 isolates being resistant to virtually all antimicrobials used in veterinary medicine is alarming and should be considered a central issue considering that MRSP ST71 spreads over large geographic distances and its transmission from animals to humans.

Characterizing biofilm formation of Staphylococcus pseudintermedius in different suture materials.

Staphylococcus pseudintermedius is the primary cause of pyoderma and surgical site infection (SSI) in dogs, and biofilm formation is the main reason for persistent SSI. The presence of biofilm in medical devices can directly impact treatment. Methicillin-resistant S. pseudintermedius (MRSP) emerged rapidly in companion animals, limiting treatment options. MRSP is a public health problem since zoonotic transmission can occur. The study seeks to evaluate biofilm formation capacity via Staphylococcus pseudintermedius collected from dogs affected by topical infections, in suture materials commonly used in companion animal surgery. We tested segments of four types of sutures. Biofilm production was measured by staining with safranin and colorimetric absorbance measurement. We calculated colony-forming units (CFUs) for each type of sutures and visualized biofilm via Scanning Electron Microscopy (SEM) images. The genes associated with biofilm formation (icaA and icaD) were identified using PCR. The colorimetric tests showed that the biofilm is most abundantly formed on the cotton sutures and polyglactin 910. The ability to form biofilm on polypropylene and nylon sutures has also been demonstrated, although at varying intensities. PCR revealed the presence of the two genes (icaA and icaD) in all the isolates. We used a positive control using a reference strain and negative control without bacteria for comparisons. Suture material allowing biofilm formation makes it difficult to prevent and treat surgical site infections. Therefore, it is important to know which suture thread is more susceptible to biofilm formation by bacteria to prevent possible secondary infections at surgical sites.

Characterization and antimicrobial susceptibility of coagulase-positive Staphylococcus isolated in a veterinary teaching hospital in Chile / Caracterización y sensibilidad antimicrobiana de Staphylococcus coagulasa-positivos aislados en un hospital clínico veterinario en Chile

Abstract There is limited information about the prevalence and antimicrobial susceptibility of coagulase-positive Staphylococcus (CoPS) strains in veterinary settings in Chile. The aim of this observational study was to identify and characterize CoPS strains from dogs, owners, veterinary professionals and surfaces in a veterinary teaching hospital at Universidad de Chile to determine the presence of methicillin-resistant strains and evaluate the genetic relationship among the strains. Veterinarians (n = 24), surfaces (n = 10), and healthy dogs (n = 40) and their respective owners (n = 40) were sampled for CoPS. Isolates were identified by PCRand antimicrobial susceptibility was assessed by the disk diffusion method and MIC. The presence of the mecA gene was evaluated by PCR, and the genetic relationship among the strains was established by PFGE. A total of 45 CoPS strains were obtained, eight from veterinary professionals, three from hospital surfaces, eight from owners and 26 from dogs. Nine of the strains were resistant to methicillin (20%), and all of them carried the mecA gene. A high percentage of the strains was resistant to clindamycin (33.3%). Additionally, the isolated CoPS showed high genetic diversity. This study suggests that veterinarians are in high risk of harboring methicillin-resistant CoPS (25% versus 2.5% from owners) and our results provide evidence that clindamycin could not be an empiric alternative for CoPS in the analyzed hospital. This is the first report of methicillin-resistant CoPS in veterinary settings in Chile, considering humans, pets and surfaces.

Resumen Existe información limitada sobre prevalencia y sensibilidad antimicrobiana de cepas de Staphylococcus coagulasa-positivas (CoPS) en entornos veterinarios en Chile. El objetivo de este estudio observacional fue identificar y caracterizar cepas CoPS de perros, duenos, veterinarios y superficies de un hospital veterinario de la Universidad de Chile, determinar la presencia de cepas meticilino-resistentes y evaluar la relación genética entre las cepas. Se colectaron muestras de veterinarios (n = 24), de superficies hospitalarias (n = 10) y de perros sanos (n =40) y sus respectivos duenos (n = 40). Los aislamientos se identificaron mediante PCR y la sensibilidad antimicrobiana se evaluó por difusión en discos y CIM. También se empleó PCR para detectar la presencia del gen mecA; la relación genética entre las cepas se estableció mediante electroforesis de campos pulsantes (PFGE). Se obtuvo un total de 45 cepas de CoPS, 8 de veterinarios, 3 de superficies hospitalarias, 8 de duenos y 26 de perros. Nueve cepas fueron meticilino-resistentes (20%), todas portadoras del gen mecA. Un porcentaje importante de cepas fue resistente a clindamicina (33,3%). Además, las cepas aisladas mostraron una alta diversidad genética. Este estudio sugiere que los veterinarios tienen alto riesgo de portar CoPS resistentes a meticilina (25% versus 2,5% propietarios). Asimismo, nuestros resultados proporcionan evidencia de que la clindamicina podría no ser una alternativa empírica para CoPS en el hospital analizado. Este es el primer estudio de CoPS meticilino-resistentes en entornos veterinarios en Chile que considera humanos, mascotas y superficies.

Characterization and antimicrobial susceptibility of coagulase-positive Staphylococcus isolated in a veterinary teaching hospital in Chile.

There is limited information about the prevalence and antimicrobial susceptibility of coagulase-positive Staphylococcus (CoPS) strains in veterinary settings in Chile. The aim of this observational study was to identify and characterize CoPS strains from dogs, owners, veterinary professionals and surfaces in a veterinary teaching hospital at Universidad de Chile to determine the presence of methicillin-resistant strains and evaluate the genetic relationship among the strains. Veterinarians (n=24), surfaces (n=10), and healthy dogs (n=40) and their respective owners (n=40) were sampled for CoPS. Isolates were identified by PCR and antimicrobial susceptibility was assessed by the disk diffusion method and MIC. The presence of the mecA gene was evaluated by PCR, and the genetic relationship among the strains was established by PFGE. A total of 45 CoPS strains were obtained, eight from veterinary professionals, three from hospital surfaces, eight from owners and 26 from dogs. Nine of the strains were resistant to methicillin (20%), and all of them carried the mecA gene. A high percentage of the strains was resistant to clindamycin (33.3%). Additionally, the isolated CoPS showed high genetic diversity. This study suggests that veterinarians are in high risk of harboring methicillin-resistant CoPS (25% versus 2.5% from owners) and our results provide evidence that clindamycin could not be an empiric alternative for CoPS in the analyzed hospital. This is the first report of methicillin-resistant CoPS in veterinary settings in Chile, considering humans, pets and surfaces.

Interspecies transfer of plasmid-borne gentamicin resistance between Staphylococcus isolated from domestic dogs to Staphylococcus aureus.

As preconized by the One Health concept, the intimate relationship between pets and owners is a common source for the trade of microorganisms with zoonotic potential, and with them, antimicrobial resistance genes. In this work, we evaluated the presence of antimicrobial resistance genes, that are usually within mobile genetic elements, in a laboratory collection of 79 canine Staphylococcus strains, mostly Staphylococcus pseudintermedius and Staphylococcus coagulans. Resistance to tetracycline was observed in 34% of the strains, followed by resistance to erythromycin (21%) and gentamicin (19%). These phenotypes were partially correlated with the presence of the tetracycline resistance genes tet(M) and tet(K) in 64% and 44% of all strains, respectively; erythromycin resistance genes erm(A) and erm(C) in 53% and 23%; and gentamicin resistance gene aac(6')-aph(2″) in 26% of the strains. At least 45% of the strains harbored high- and/or low-molecular weight plasmids, whose transfer may be facilitated by their widespread biofilm-forming capacity, and absence of restrictive CRISPR systems. We selected eight plasmid-bearing and multidrug resistant strains, which were submitted to plasmid curing by stress with SDS. No strain lost resistance during stressing cultivation but, by conjugation experiments, the S. pseudintermedius strain 27 transferred its plasmid-borne resistance to gentamicin, conferred by the aac(6')-aph(2″) gene, to Staphylococcus aureus. The frequent empirical use of gentamicin to treat skin and ear infections in domestic dogs is likely to select resistant strains. Also, as demonstrated by our study, these strains can serve as gene reservoirs for human pathogens, such as S. aureus.

A fatal pneumonia due to coinfection of Pseudomonas putida and Staphylococcus pseudintermedius in a laboratory beagle dog

Background: Pseudomonas putida (P. putida) is widely distributed in the environment, and sometimes caused nosocomialinfections in human beings, but no case of infection has been reported in beagle dogs. Staphylococcus pseudintermedius(S. pseudintermedius) is a natural cutaneous bacterium in dogs and occasionally causes purulent infections of the skin yetrarely causes pneumonia. Both bacteria are opportunistic pathogens. Dogs, even well-controlled laboratory beagle dogs,maybe infected by the bacterium in certain conditions like this report. In order to provide information and give suggestionto veterinarians involved in dogs study, a complete profile of the coinfection was drawn in this report.Case: It is presented a case of an 8-month-old beagle dog, weighing 6 kg that suffered from coinfection of P. putida andS. pseudintermedius during a treatment of chemotherapy. The animal was confirmed as normal by appearance, physicalexamination and laboratory tests before arrival according to the applicable guidelines. After 14-day acclimation period, theanimal was administrated with a tyrosinase inhibitor once daily via oral gavage. From Day 8, coughing, decreased activity, hyporeflexia, squinting, shortness of breath (abdominal breathing), and discharge around the nose as well as cracklesin the lung and rapid heart rate were noted. Since the poor conditions progressed quickly and have not been improved bytreatment of ceftriaxone and dexamethasone. On Day 9, the animal was euthanized for humanitarian reasons. To define thepathogen, hilar lymph node and thoracic swab were collected for bacteria isolation and purification in special mediums,and at last characterized by Gram staining and 16s rRNA gene sequence analysis and positive PCR-restriction fragmentlength polymorphism. In clinical pathological examination, an increase in WBC, neutrophils, lymphocytes, monocytes...(AU)

A fatal pneumonia due to coinfection of Pseudomonas putida and Staphylococcus pseudintermedius in a laboratory beagle dog

Background: Pseudomonas putida (P. putida) is widely distributed in the environment, and sometimes caused nosocomialinfections in human beings, but no case of infection has been reported in beagle dogs. Staphylococcus pseudintermedius(S. pseudintermedius) is a natural cutaneous bacterium in dogs and occasionally causes purulent infections of the skin yetrarely causes pneumonia. Both bacteria are opportunistic pathogens. Dogs, even well-controlled laboratory beagle dogs,maybe infected by the bacterium in certain conditions like this report. In order to provide information and give suggestionto veterinarians involved in dogs study, a complete profile of the coinfection was drawn in this report.Case: It is presented a case of an 8-month-old beagle dog, weighing 6 kg that suffered from coinfection of P. putida andS. pseudintermedius during a treatment of chemotherapy. The animal was confirmed as normal by appearance, physicalexamination and laboratory tests before arrival according to the applicable guidelines. After 14-day acclimation period, theanimal was administrated with a tyrosinase inhibitor once daily via oral gavage. From Day 8, coughing, decreased activity, hyporeflexia, squinting, shortness of breath (abdominal breathing), and discharge around the nose as well as cracklesin the lung and rapid heart rate were noted. Since the poor conditions progressed quickly and have not been improved bytreatment of ceftriaxone and dexamethasone. On Day 9, the animal was euthanized for humanitarian reasons. To define thepathogen, hilar lymph node and thoracic swab were collected for bacteria isolation and purification in special mediums,and at last characterized by Gram staining and 16s rRNA gene sequence analysis and positive PCR-restriction fragmentlength polymorphism. In clinical pathological examination, an increase in WBC, neutrophils, lymphocytes, monocytes...

Molecular Detection and Characterization of the mecA and nuc Genes From Staphylococcus Species (S. aureus, S. pseudintermedius, and S. schleiferi) Isolated From Dogs Suffering Superficial Pyoderma and Their Antimicrobial Resistance Profiles.

Canine superficial pyoderma (CSP) is a bacterial infection secondary to several skin diseases of the dog. Staphylococcus pseudintermedius, which is a commensal bacterium of the dog's skin, is the leading agent found in dogs affected by CSP, which can progress to deep pyoderma. It is also of clinical significance because S. pseudintermedius strains carry antimicrobial resistance genes, mainly the mecA gene. In this descriptive longitudinal study, molecular characterization of bacterial isolates from dogs affected by CSP was performed in addition to phenotyping, antimicrobial profiling, and assessment of resistance carriage status. Fifty dogs (24 females and 26 males) attending the CES University Veterinary Teaching Hospital were included in the study. CSP was confirmed according to clinical signs and cytological examination. Swabs were taken from active skin lesions for bacterial culture, and phenotyping and antimicrobial resistance profiles were assessed using API-Staph phenotyping and the Kirby-Bauer method, respectively. We also performed molecular detection and characterization of the mecA and nuc encoding gene of coagulase-positive Staphylococci. The mecA gene frequency was established by qPCR amplification of a 131bp gene fragment. Data were evaluated by descriptive statistics. Erythema, peeling, pruritus, and alopecia were the predominant symptoms (72, 56, and 46%, respectively). We isolated bacteria compatible with Staphylococcus species from all samples tested. API phenotyping showed 83.1 to 97.8% compatibility with S. pseudintermedius. PCR-genotyping resulted in 15, 3, and 1 isolates positive for S. pseudintermedius, S. aureus, and S. schleiferi, respectively. Isolated strains showed high susceptibility to Imipenem, Ampicillin/Sulbactam, and Rifampicin (100, 94, and 92%, respectively). The highest resistance was against Vancomycin and Trimethoprim/Sulfamethoxazole (98 and 74%, respectively). S. pseudintermedius, S. aureus, and S. schleiferi isolates were cloned and shared 96% sequence homology. Finally, we found 62% carriage status of the mecA gene in isolates of CSP patients, although only 36% of the isolates were methicillin-resistant. Identification of three Staphylococcus species causing CSP, high-level resistance against conventional antimicrobials, and carriage of the mecA gene highlight the importance of performing molecular characterization of bacteria causing dermatological conditions in dogs.

Characterization of the First mecA-Positive Multidrug-Resistant Staphylococcus pseudintermedius Isolated from an Argentinian Patient.

Staphylococcus pseudintermedius is commonly associated with colonization or infection in dogs, and was identified as a novel species within the genus Staphylococcus in 2006. Methicillin resistance emerged in S. pseudintermedius during the last decade. We describe here a genomic characterization of the first methicillin-resistant S. pseudintermedius (MRSP) recovered from a human patient in Argentina. The strain was phenotypically identified as MRSP 8510 by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) and antimicrobial susceptibility testing. We assessed genetic characterization by mecA PCR, SCCmec (staphylococcal chromosomal cassette) typing, and whole-genome sequencing. MRSP 8510 was phenotypically resistant to six classes of antimicrobial agents, consistent with the genes found in its genome. We concluded that MRSP 8510 was a multidrug-resistant ST1412 isolate. This study highlights the importance of the detection and characterization of pathogens with potential risks of zoonotic transmission to humans, as they may constitute a reservoir of genes associated with antimicrobial resistance.

Ceftaroline resistance in Staphylococcus pseudintermedius gene mecA carriers / Resistência à ceftarolina em Staphylococcus pseudintermedius portadores do gene mecA

Infections caused by methicillin-resistant Staphylococcus aureus (MRSA) being a constant concern, ceftaroline fosamil has been recently approved as a new cephalosporin, active against MRSA, for use in humans; only rare cases of resistance have been reported till date. There is no report of resistance to ceftaroline in Staphylococcus pseudintermedius, which is the main bacterium causing dermatitis and otitis in dogs. To evaluate staphylococcal resistance to ceftaroline, 35 isolates of methicillin-resistant S. pseudintermedius (MRSP), carrying the mecA gene, from 26 dogs with folliculitis and nine dogs with external otitis, underwent disk diffusion test with cefoxitin, oxacillin, and ceftaroline. Tests with cefoxitin and oxacillin showed > 90% sensitivity in methicillin resistance detection. In the disk diffusion test, 97.14% (34/35) were resistant to cefoxitin, 94.29% (33/35) to oxacillin, and 31.43% (11/35) to ceftaroline. Of the ceftaroline-resistant strains, 27.27% (3/11) were obtained from the ears of dogs while the rest (8/11) were from the skin. The current report is the first description of MRSP resistance to ceftaroline.(AU)

Infecções causadas por Staphylococcus aureus resistente à meticilina (MRSA) são uma preocupação médica constante. A ceftarolina fosamila é uma nova cefalosporina ativa contra Staphylococcus aureus resistente à meticilina recentemente aprovada para uso em humanos e raros casos de resistência relatados até agora. Não há relatos de resistência à ceftarolina em Staphylococcus pseudintermedius, principal bactéria causadora de dermatite e otite em cães. Com o objetivo de avaliar a resistência estafilocócica à ceftarolina, 35 amostras de S. pseudintermedius resistentes à meticilina (MRSP), portadoras do gene mecA, provenientes de 26 cães com foliculite e 9 com otite externa foram submetidos ao teste de disco-difusão com cefoxitina, oxacilina e ceftarolina. Os testes realizados com cefoxitina e oxacilina mostraram mais de 90% de sensibilidade na detecção da resistência à meticilina em ambas. No teste da disco-difusão, 97,14% (1/35) foram resistentes à cefoxitina, 94,29% (3/35) à oxacilina e 31,43% (11/35) à ceftarolina. Das cepas resistentes às ceftarolina, 27,27 (3/11) foram provenientes de ouvido de cães e as demais (8/11), provenientes da pele, sendo essa primeira descrição de resistência de MRSP à ceftarolina na literatura atual.(AU)

Ceftaroline resistance in Staphylococcus pseudintermedius gene mecA carriers / Resistência à ceftarolina em Staphylococcus pseudintermedius portadores do gene mecA

Infections caused by methicillin-resistant Staphylococcus aureus (MRSA) being a constant concern, ceftaroline fosamil has been recently approved as a new cephalosporin, active against MRSA, for use in humans; only rare cases of resistance have been reported till date. There is no report of resistance to ceftaroline in Staphylococcus pseudintermedius, which is the main bacterium causing dermatitis and otitis in dogs. To evaluate staphylococcal resistance to ceftaroline, 35 isolates of methicillin-resistant S. pseudintermedius (MRSP), carrying the mecA gene, from 26 dogs with folliculitis and nine dogs with external otitis, underwent disk diffusion test with cefoxitin, oxacillin, and ceftaroline. Tests with cefoxitin and oxacillin showed > 90% sensitivity in methicillin resistance detection. In the disk diffusion test, 97.14% (34/35) were resistant to cefoxitin, 94.29% (33/35) to oxacillin, and 31.43% (11/35) to ceftaroline. Of the ceftaroline-resistant strains, 27.27% (3/11) were obtained from the ears of dogs while the rest (8/11) were from the skin. The current report is the first description of MRSP resistance to ceftaroline.(AU)

Infecções causadas por Staphylococcus aureus resistente à meticilina (MRSA) são uma preocupação médica constante. A ceftarolina fosamila é uma nova cefalosporina ativa contra Staphylococcus aureus resistente à meticilina recentemente aprovada para uso em humanos e raros casos de resistência relatados até agora. Não há relatos de resistência à ceftarolina em Staphylococcus pseudintermedius, principal bactéria causadora de dermatite e otite em cães. Com o objetivo de avaliar a resistência estafilocócica à ceftarolina, 35 amostras de S. pseudintermedius resistentes à meticilina (MRSP), portadoras do gene mecA, provenientes de 26 cães com foliculite e 9 com otite externa foram submetidos ao teste de disco-difusão com cefoxitina, oxacilina e ceftarolina. Os testes realizados com cefoxitina e oxacilina mostraram mais de 90% de sensibilidade na detecção da resistência à meticilina em ambas. No teste da disco-difusão, 97,14% (1/35) foram resistentes à cefoxitina, 94,29% (3/35) à oxacilina e 31,43% (11/35) à ceftarolina. Das cepas resistentes às ceftarolina, 27,27 (3/11) foram provenientes de ouvido de cães e as demais (8/11), provenientes da pele, sendo essa primeira descrição de resistência de MRSP à ceftarolina na literatura atual.(AU)

An investigation of methicillin-resistant Staphylococcus pseudintermedius (MRSP) in domestic and shelter dogs in Montenegro (RS-Brazil) / Pesquisa de “Staphylococcus pseudintermedius” resistente à meticilina (MRSP) em cães domésticos e de abrigo em Montenegro (RS-Brasil)

The interaction established between man and their pets has been increasing progressively. The diagnosis of animals carrying methicillin-resistant Staphylococcus pseudintermedius (MRSP) becomes important since positive results for this opportunistic bacteria has been found in humans. The aim of this study was to investigate methicillin-resistant Staphylococcus pseudintermedius (MRSP) in domestic dogs attended in a veterinary clinic and shelter dogs. Anal and nasal swabs from 53 domestic dogs and 46 shelter dogs were collected between the months of August and October of 2015, resulting in 198 samples, which were sent for routine laboratory analysis and subsequently used for this study. Phenotypic tests for bacteria identification and susceptibility to oxacillin were performed for the investigation of MRSP. A total of 23 samples (11.6%) from 21 dogs were positive for S. pseudintermedius, however none of the isolates were MRSP. Nasal and anal swabs had similar positivity and two dogs were carriers of the bacteria in both analyzed sites (anal and nasal). Our results showed a higher positivity for S. pseudintermedius in domestic dogs compared to shelter dogs, as well as indicating the importance of collecting more than one colonization site. This is the first study to perform a comparison among different animal creating sites in Brazil.(AU)

O convívio estabelecido entre o homem e seus animais de estimação vem se intensificando progressivamente. O diagnóstico de animais portadores de Staphylococcus pseudintermedius resistente à meticilina (MRSP) se torna importante uma vez que já foram obtidos resultados positivos para esta bactéria oportunista em humanos. O objetivo do estudo foi pesquisar Staphylococcus pseudintermedius resistente à meticilina (MRSP) em cães domésticos atendidos numa clínica veterinária e em cães de abrigo. Entre os meses de Agosto e Outubro de 2015 foram coletados swabs anais e nasais de 53 cães domésticos e de 46 cães de abrigo, resultando em um total de 198 amostras, as quais foram enviadas para análise de rotina laboratorial na Universidade Feevale e, posteriormente, utilizadas para este estudo. Foram realizados testes fenotípicos de identificação da bactéria e de suscetibilidade à oxacilina para a pesquisa de MRSP. Um total de 23 amostras (11.6%) oriundas de 21 cães foi positivo para S. pseudintermedius, porém nenhum isolado era MRSP. Os swabs nasais e anais tiveram positividade semelhante, sendo dois cães portadores da bactéria em ambos os sítios (anal e nasal). Nossos resultados indicaram uma maior positividade para S. pseudintermedius em cães domésticos em comparação aos cães de abrigo, além de sinalizar sobre a importância de coletas em mais de um sítio de colonização. Este é o primeiro estudo que realizou uma comparação entre os diferentes locais de criação dos animais no Brasil.(AU)

An investigation of methicillin-resistant Staphylococcus pseudintermedius (MRSP) in domestic and shelter dogs in Montenegro (RS-Brazil) / Pesquisa de “Staphylococcus pseudintermedius” resistente à meticilina (MRSP) em cães domésticos e de abrigo em Montenegro (RS-Brasil)

The interaction established between man and their pets has been increasing progressively. The diagnosis of animals carrying methicillin-resistant Staphylococcus pseudintermedius (MRSP) becomes important since positive results for this opportunistic bacteria has been found in humans. The aim of this study was to investigate methicillin-resistant Staphylococcus pseudintermedius (MRSP) in domestic dogs attended in a veterinary clinic and shelter dogs. Anal and nasal swabs from 53 domestic dogs and 46 shelter dogs were collected between the months of August and October of 2015, resulting in 198 samples, which were sent for routine laboratory analysis and subsequently used for this study. Phenotypic tests for bacteria identification and susceptibility to oxacillin were performed for the investigation of MRSP. A total of 23 samples (11.6%) from 21 dogs were positive for S. pseudintermedius, however none of the isolates were MRSP. Nasal and anal swabs had similar positivity and two dogs were carriers of the bacteria in both analyzed sites (anal and nasal). Our results showed a higher positivity for S. pseudintermedius in domestic dogs compared to shelter dogs, as well as indicating the importance of collecting more than one colonization site. This is the first study to perform a comparison among different animal creating sites in Brazil.

O convívio estabelecido entre o homem e seus animais de estimação vem se intensificando progressivamente. O diagnóstico de animais portadores de Staphylococcus pseudintermedius resistente à meticilina (MRSP) se torna importante uma vez que já foram obtidos resultados positivos para esta bactéria oportunista em humanos. O objetivo do estudo foi pesquisar Staphylococcus pseudintermedius resistente à meticilina (MRSP) em cães domésticos atendidos numa clínica veterinária e em cães de abrigo. Entre os meses de Agosto e Outubro de 2015 foram coletados swabs anais e nasais de 53 cães domésticos e de 46 cães de abrigo, resultando em um total de 198 amostras, as quais foram enviadas para análise de rotina laboratorial na Universidade Feevale e, posteriormente, utilizadas para este estudo. Foram realizados testes fenotípicos de identificação da bactéria e de suscetibilidade à oxacilina para a pesquisa de MRSP. Um total de 23 amostras (11.6%) oriundas de 21 cães foi positivo para S. pseudintermedius, porém nenhum isolado era MRSP. Os swabs nasais e anais tiveram positividade semelhante, sendo dois cães portadores da bactéria em ambos os sítios (anal e nasal). Nossos resultados indicaram uma maior positividade para S. pseudintermedius em cães domésticos em comparação aos cães de abrigo, além de sinalizar sobre a importância de coletas em mais de um sítio de colonização. Este é o primeiro estudo que realizou uma comparação entre os diferentes locais de criação dos animais no Brasil.