Methicillin-susceptible S. Pseudintermedius Research Articles

Rapid and visual RPA-CRISPR/Cas12a detection for Staphylococcus pseudintermedius and methicillin-resistant S. pseudintermedius in clinical samples of dogs and cats

Staphylococcus pseudintermedius can cause severe infections of the skin, ear and other tissues in dogs and cats. Methicillin-resistant S. pseudintermedius (MRSP) has recently become more prevalent, posing a severe threat to companion animals and public health. Therefore, rapid and accurate diagnosis of S. pseudintermedius and MRSP infections in dogs and cats is essential for timely controlling infections. The development of CRISPR/Cas technology offers an innovative solution for rapid diagnosis. Here, we established an assay combining recombinant polymerase amplification (RPA) and CRISPR/Cas12a. By separately detecting the spsJ gene, the specific gene of S. pseudintermedius, and the mecA gene, the methicillin resistance gene, this method allows for the direct detection of methicillin-susceptible S. pseudintermedius (MSSP) and MRSP in clinical samples at 37 °C for a total of 40 min, The results can be directly visualized by the naked eye under blue light. The limits of detection of the RPA-CRISPR/Cas12a assay were 103 copies per reaction for the spsJ gene and 104 copies per reaction for the mecA gene. The RPA-CRISPR/Cas12a detection successfully detected and differentiated clinical isolates of MSSP and MRSP without cross-reactivity with other tested bacteria species. The evaluation of the detection performance of RPA-CRISPR/Cas12a with 47 clinical samples (without culture) from dogs and cats showed that the results of detection were 100% consistent with those of clinical culture and colony sequencing, which was more sensitive than PCR. RPA-CRISPR/Cas12a assay can quickly and sensitively detect S. pseudintermedius and MRSP in clinical samples without expensive instruments, making it suitable for small veterinary clinics.

Comparative Susceptibility of Pathogenic Methicillin-Resistant and Methicillin-Susceptible Staphylococcus pseudintermedius to Empirical Cotrimazole for Canine Pyoderma.

The prevalence of methicillin-resistant Staphylococcus pseudintermedius (MRSP) that causes pyoderma has been gradually shifting, according to many surveillance studies, with annual changes. The empirical cotrimazole regimen remains interesting, but research on cotrimazole susceptibility to MRSP is limited. The objective of this study was to evaluate the susceptibility of cotrimazole to canine pyoderma MRSP isolates. Sixty isolates of S. pseudintermedius were identified as 16 MRSP and 44 methicillin-susceptible S. pseudintermedius (MSSP) using an oxacillin disk diffusion test and VITEK 2 system with VITEK GP card. Using the VITEK 2 system with a VITEK AST-GP81 card, the susceptibility rates of MRSP (15.00%) and MSSP (35.00%) to cotrimazole was observed. The median MIC of cotrimazole on MSSP (median, ≤10; IQR, 10-320) was lower than that of MRSP (median, ≥320; IQR, 10-320) (p = 0.5889, Mann-Whitney test). Percent attainment of PK/PD targets in MRSP (q 12 h, 43.75; q 8 h, 43.75) were lower than that of MSSP (q 12 h, 52.27; q 8 h, 52.27) (p = 0.7710). These findings show the moderately phenotypic cotrimazole susceptibilities of both MRSP and MSSP. Further study is required to develop clinical trials examining the use of cotrimazole in dogs with pyoderma.

In Vitro Antimicrobial Activity of Piper betle Leaf Extract and Some Topical Agents against Methicillin-Resistant and Methicillin-Susceptible Staphylococcus Strains from Canine Pyoderma.

As multidrug-resistant methicillin-resistant staphylococci (MRS) is becoming more prevalent in canine pyoderma, the discovery of new therapeutic options is required. This study aimed to test the antimicrobial activity of crude Piper betle leaf extract and some topical antimicrobial agents against canine Staphylococcus clinical strains by determining the minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC). The results showed that the mean MICs of chlorhexidine, miconazole, crude P. betle leaf extract, azelaic acid, and benzoyl peroxide against Staphylococcus strains were 1.41, 1.62, 252.78, 963.49, and 1342.70 mg/L, respectively. Therefore, betel leaf extract demonstrated a superior efficacy to azelaic acid and benzoyl peroxide. Furthermore, the ratio of MBC/MIC of betel leaf extract was 1.75, indicating its bactericidal action. When applied to methicillin-resistant S. pseudintermedius (MRSP) and methicillin-susceptible S. pseudintermedius (MSSP), betel leaf extract was equally efficient towards both groups. S. pseudintermedius strains were more susceptible to betel leaf extract than S. schleiferi subsp. coagulans. In gas chromatography-mass spectrometry analysis, eugenol and hydroxychavicol appeared to be the major components of betel leaf extract. Given its efficacy, dogs with pyoderma could benefit from the use of betel leaf extract as a topical antimicrobial alternative.

Comparison of the prevalence, antibiotic resistance patterns, and biofilm formation ability of methicillin-resistant Staphylococcus pseudintermedius in healthy dogs and dogs with skin infections.

Staphylococcus pseudintermedius (S. pseudintermedius), found on dogs' skin and mucous membranes, can act as an opportunistic pathogen causing skin, ear, and other tissue infections. Due to the possibility of zoonotic transmission of them, it is necessary to investigate the prevalence of S. pseudintermedius, especially the antimicrobial-resistant strains, in pets. In this study, the prevalence, antimicrobial resistance patterns, and biofilm formation ability of methicillin-resistant S. pseudintermedius (MRSP) were investigated and compared in 50 healthy dogs and 50 dogs with skin infections. The prevalence of S. pseudintermedius was not significantly different between healthy dogs (40%) and dogs with skin infections (50%). No significant difference was found in the prevalence of MRSP between healthy dogs (12%) and dogs with skin infections (18%). A total of 81.8% of S. pseudintermedius isolates were biofilm producers. The frequencies of antibiotic resistance (except for gentamicin), multidrug resistance (MDR), and biofilm formation ability were not significantly different between S. pseudintermedius isolates of healthy dogs and dogs with skin infections. The frequencies of resistance to penicillin and tetracycline, MDR, and biofilm production abilities were significantly higher among MRSP than methicillin-susceptible S. pseudintermedius (MSSP). The frequency of oxacillin resistance was significantly higher among weak or moderate biofilm producers than non-biofilm producers. The frequency of resistance to erythromycin was significantly higher among moderate biofilm producers than non-biofilm producers or weak biofilm producers. High frequencies of biofilm-producer S. pseudintermedius isolates and their resistance to antibiotics can affect the success of treatment of infections caused by these strains.

Transmission of Methicillin-Resistant Staphylococcus spp. from Infected Dogs to the Home Environment and Owners.

Dogs with methicillin-resistant Staphylococcus spp. (MRS) infections often undergo treatment in their homes, interacting with their owners and surroundings. This close contact between dogs and owners may facilitate the interspecies transmission of MRS. Therefore, this study aimed to investigate the transmission of MRS from infected dogs to their owners and home environments. Seven households with dogs that had been diagnosed with methicillin-resistant S. pseudintermedius (MRSP) and one household with a dog with methicillin-resistant S. epidermidis (MRSE) participated in the study. Dogs, owners, and the home environments were screened for the presence of clinical MRS. A selection of 36 staphylococcal isolates were whole-genome sequenced and screened for resistance genes and virulence genes. Clinical MRS were primarily identified from the dogs and their immediate surroundings, but these were also detected in locations that were out of reach for the dogs, indicating indirect transmission. Two of eight owners carried clinical MRS in their nostrils, while one owner carried methicillin-susceptible S. pseudintermedius (MSSP). All clinical MRS were multi-resistant, and several possessed resistance genes that were not expressed phenotypically. Clinical MRSP persisted in the home environment for a prolonged period, despite infection recovery and one dog being euthanized. Regardless of the stable presence of MRSP in the surroundings, the owners in these homes remained negative, but tested positive for MSSP on three occasions.

An overview of human infections caused by <em>Staphylococcus pseudintermedius:</em> A zoonotic risk of the oldest friend

Staphylococcus pseudintermedius is colonized in 90% of healthy dogs and the infection has also been reported in humans who have close contact with dogs. S. pseudintermedius is a Gram positive coagulase positive coccus which is an opportunistic pathogen. Human infection has been reported less frequently either due to misidentification or misdiagnosis or to underreporting. The objective of this overview is to highlight the epidemiology, clinical presentation, importance of laboratory tests and antimicrobial resistance of S. pseudintermedius in humans.Human infections of S. pseudintermedius have been reported as case studies or clusters of infections in the literature. Several cases have been reported among the adult population, and in patients with metabolic diseases such as diabetes mellitus, immunocompromised states such as AIDS, cancers, and autoimmune diseases. The organism had been isolated and identified both in healthy people and individuals with adverse clinical outcomes. S. pseudintermedius causes skin and soft tissue infections, catheter associated infections and surgical site infections in humans. The incidence of skin and soft tissue infection by S. pseudintermedius in humans was 0.025% in Canada. Methicillin resistance S. pseudintermedius (MRSP) has been isolated both in healthy people and associated with clinical infections. MRSP is often multidrug resistant and therapeutic options are limited. Multidrug resistance has been detected in methicillin susceptible S. pseudintermedius (MSSP). Close association of a canine host is the major risk factor in humans and vulnerable individuals need to be persuaded to practice hygienic practices to minimize the risk of zoonotic infection.In conclusion, differentiation of S. pseudintermedius from S. aureus is required to understand the epidemiology of the disease, diagnosis, clinical presentation, and further interpretation of antimicrobial resistance in the human clinical context. Suitable hygienic practices post handling of dogs and improvement of laboratory facilities would help minimize the incidence of S. pseudintermedius infections in humans.

Absence of Host-Specific Genes in Canine and Human Staphylococcus pseudintermedius as Inferred from Comparative Genomics.

Staphylococcus pseudintermedius is an important pathogen in dogs that occasionally causes infections in humans as an opportunistic pathogen of elderly and immunocompromised people. This study compared the genomic relatedness and antimicrobial resistance genes using genome-wide association study (GWAS) to examine host association of canine and human S. pseudintermedius isolates. Canine (n = 25) and human (n = 32) methicillin-susceptible S. pseudintermedius (MSSP) isolates showed a high level of genetic diversity with an overrepresentation of clonal complex CC241 in human isolates. This clonal complex was associated with carriage of a plasmid containing a bacteriocin with cytotoxic properties, a CRISPR-cas domain and a pRE25-like mobile element containing five antimicrobial resistance genes. Multi-drug resistance (MDR) was predicted in 13 (41%) of human isolates and 14 (56%) of canine isolates. CC241 represented 54% of predicted MDR isolates from humans and 21% of predicted MDR canine isolates. While it had previously been suggested that certain host-specific genes were present the current GWAS analysis did not identify any genes that were significantly associated with human or canine isolates. In conclusion, this is the first genomic study showing that MSSP is genetically diverse in both hosts and that multidrug resistance is important in dog and human-associated S. pseudintermedius isolates.

Genes on the Move: In Vitro Transduction of Antimicrobial Resistance Genes between Human and Canine Staphylococcal Pathogens.

Transmission of methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-resistant Staphylococcus pseudintermedius (MRSP) between people and pets, and their co-carriage, are well-described. Potential exchange of antimicrobial resistance (AMR) genes amongst these staphylococci was investigated in vitro through endogenous bacteriophage-mediated transduction. Bacteriophages were UV-induced from seven donor isolates of canine (MRSP) and human (MRSA) origin, containing tet(M), tet(K), fusB or fusC, and lysates filtered. Twenty-seven tetracycline- and fusidic acid- (FA-) susceptible recipients were used in 122 donor-recipient combinations (22 tetracycline, 100 FA) across 415 assays (115 tetracycline, 300 FA). Bacteriophage lysates were incubated with recipients and presumed transductants quantified on antimicrobial-supplemented agar plates. Tetracycline resistance transduction from MRSP and MRSA to methicillin-susceptible S. pseudintermedius (MSSP) was confirmed by PCR in 15/115 assays. No FA-resistance transfer occurred, confirmed by negative fusB/fusC PCR, but colonies resulting from FA assays had high MICs (≥32 mg/L) and showed mutations in fusA, two at a novel position (F88L), nine at H457[Y/N/L]. Horizontal gene transfer of tetracycline-resistance confirms that resistance genes can be shared between coagulase-positive staphylococci from different hosts. Cross-species AMR transmission highlights the importance of good antimicrobial stewardship across humans and veterinary species to support One Health.

Antimicrobial Activity of Some Essential Oils against Methicillin-Susceptible and Methicillin-Resistant Staphylococcus pseudintermedius-Associated Pyoderma in Dogs.

Simple SummaryPyoderma is one of the most common diseases in dogs, and Staphylococcus pseudintermedius, a Gram-positive coagulase-positive bacterium, represents the most common infectious agent causing canine pyoderma. Since multidrug-resistant S. pseudintermedius strains have become a relevant threat in veterinary medicine, this study aimed to test the antimicrobial properties of some essential oils (EOs) against S. pseudintermedius strains isolated from dogs suffering from pyoderma. The obtained findings demonstrated a clear in vitro efficacy of some tested EOs against clinical methicillin-resistant and methicillin-sensible S. pseudintermedius strains. The applicability and efficacy of EOs in cases of canine pyoderma supported by S. pseudintermedius could be beneficial for both dogs and pet owners, who are inevitably exposed to this zoonotic bacterium.This study aimed to test in vitro the antimicrobial activity of 11 essential oils (EOs) against four methicillin-resistant Staphylococcus pseudintermedius (MRSP) and four methicillin-susceptible S. pseudintermedius (MSSP) clinical isolates. The obtained findings demonstrated a clear in vitro efficacy of some tested EOs against both MRSP and MSSP strains. Particularly, modal minimum inhibitory concentration (MIC) values ranging from 1:2048 v/v for Melissa officinalis against an MSSP strain to 1:256 v/v for Cymbopogon citratus against all MRSP strains were observed. The best results, highlighting a modal MIC value of 1:1024 v/v for all tested isolates, was provided by Cinnamomum zeylanicum. Intriguingly, Cinnamomum zeylanicum showed, in many cases, a correspondence between minimum bactericidal concentration (MBC) and MIC values, indicating that the inhibiting dose is also often bactericidal. Moreover, a mild antibacterial and bactericidal activity against both MRSP and MSSP isolates was detected for the other tested EOs. Considering the zoonotic potential of S. pseudintermedius and the increased dissemination of multidrug-resistant strains, the employment of EOs could be useful for the treatment of canine pyoderma. Since antibiotic resistance has become the most urgent issue, from the perspective of the One Health initiative, alternative therapeutic approaches are desirable to limit the use of antibiotics or to improve the efficacy of conventional therapies.

Novel Antimicrobial Peptide from Temporin L in The Treatment of Staphylococcus pseudintermedius and Malassezia pachydermatis in Polymicrobial Inter-Kingdom Infection.

Interkingdom polymicrobial diseases are caused by different microorganisms that colonize the same niche, as in the case of yeast-bacteria coinfections. The latter are difficult to treat due the absence of any common therapeutic target for their elimination, both in animals and humans. Staphylococcus pseudintermedius and Malassezia pachydermatis belong to distinct kingdoms. They can colonize the same skin district or apparatus being the causative agents of fastidious pet animals’ pathologies. Here we analysed the antimicrobial properties of a panel of 11 peptides, derived from temporin L, against Malassezia pachydermatis. Only peptide 8 showed the best mycocidal activity at 6.25 μM. Prolonged application of peptide 8 did not cause M. pachydermatis drug-resistance. Peptide 8 was also able to inhibit the growth of Staphylococcus pseudintermedius, regardless of methicillin resistance, at 1.56 μM for methicillin-susceptible S. pseudintermedius (MSSP) and 6.25 μM for methicillin-resistant S. pseudintermedius (MRSP). Of interest, peptide 8 increased the susceptibility of MRSP to oxacillin. Oxacillin MIC value reduction was of about eight times when used in combination with peptide 8. Finally, the compound affected the vitality of bacteria embedded in S. pseudintermedius biofilm. In conclusion, peptide 8 might represent a valid therapeutic alternative in the treatment of interkingdom polymicrobial infections, also in the presence of methicillin-resistant bacteria.

Geographical Variations in Virulence Factors and Antimicrobial Resistance Amongst Staphylococci Isolated From Dogs From the United Kingdom and Romania.

The objective of this study was to compare virulence and resistance factors of mucosal and cutaneous staphylococci from dogs with pyoderma in the UK and Romania, two countries with different approaches to antimicrobial use in companion animals. Staphylococcal isolates (n = 166) identified to the species level as being Staphylococcus pseudintermedius or coagulase negative (CoNS) were analyzed for their antimicrobial resistance (AMR) profile and presence of resistance and virulence genes. Of the investigated isolates, 26 were methicillin-resistant S. pseudintermedius (MRSP), 89 were methicillin-susceptible S. pseudintermedius (MSSP) and 51 were coagulase negative staphylococci (CoNS). A significantly larger number of isolates originating from Romania were resistant to clindamycin, tetracycline, and chloramphenicol compared to the UK isolates (P < 0.05). Resistance to amoxicillin-clavulanic acid, gentamicin, and trimethoprim-sulphamethoxazole was more evident in UK isolates. Fusidic acid resistance was common in Staphylococcus spp. isolates from both countries. Most isolates carried virulence factors associated with siet (exfoliative toxin) and luk (leucocidin) genes. All MRSP UK isolates exhibited fusidic acid resistance genes whilst this was very rare in the MRSP isolates from Romania. The chlorhexidine resistance gene qacA/B was frequently identified in CoNS isolates from the UK (P < 0.001). The current study documented differences in antimicrobial resistance profiles of Staphylococcus spp. isolates from dogs in two geographical locations in Europe, which could reflect differences in antimicrobial prescribing patterns. The study also highlights the need for further studies and interventions on antimicrobial use, prescribing patterns and AMR surveillance in companion animals in Romania.

Isolation and characterization of bacteriophages active against methicillin-resistant Staphylococcus pseudintermedius

We aimed to isolate and characterize bacteriophages (phages) with preferential activity against methicillin-resistant Staphylococcus pseudintermedius (MRSP), a multidrug-resistant canine pathogen. Four phages were isolated from canine faeces using two MRSP strains as initial hosts. Phage host range was evaluated by the spot test on 17 MRSP, 43 methicillin-susceptible S. pseudintermedius (MSSP), and six other staphylococci isolated from dogs. Transmission electron microscopy was used for presumptive identification followed by whole genome sequencing (WGS). All phages lysed all MRSP isolates whereas only 16–28% of MSSP were lysed. Their lytic activity was limited to S. pseudintermedius and S. schleiferi. All phages had similar morphology and belonged to the Siphoviridae family. WGS indicated that the phages were 93.8–99.7% identical to each other, and exhibited the highest similarity (87%) to the temperate S. aureus phage 187. Confirmatory lytic activity tests showed that phages were able to produce clear plaques on lysogens, which was enabled by recombination of the lysogeny modules as shown by WGS of the phages after propagation and plaque formation. This study provides insight into the genetic diversity and biology of S. pseudintermedius temperate phages, which could be further developed for topical therapy of MRSP skin and wound infections.

Susceptibility in vitro of canine methicillin-resistant and -susceptible staphylococcal isolates to fusidic acid, chlorhexidine and miconazole: opportunities for topical therapy of canine superficial pyoderma.

ObjectivesIncreasing multidrug resistance amongst canine pathogenic staphylococci has renewed interest in topical antibacterial therapy for skin infections in the context of responsible veterinary prescribing. We therefore determined the activity in vitro of three clinically relevant topical agents and synergism between two of them against Staphylococcus pseudintermedius and Staphylococcus aureus.MethodsThe MICs of fusidic acid (n = 199), chlorhexidine (n = 198), miconazole (n = 198) and a 1:1 combination of miconazole/chlorhexidine (n = 198) were determined for canine isolates [50 MRSA and 49 methicillin-resistant S. pseudintermedius (MRSP), 50 MSSA and 50 methicillin-susceptible S. pseudintermedius (MSSP)] collected from the UK and Germany using an agar dilution method (CLSI VET01-A4). Fractional inhibitory concentration (FIC) indices were calculated to assess the interaction of miconazole with chlorhexidine.ResultsMICs of each drug/combination were significantly (P < 0.0005) higher for S. aureus when compared with S. pseudintermedius. Most strains (n = 172) had an MIC of fusidic acid of ≤0.03 mg/L (MIC ≥64 mg/L, n = 5 MRSA). All strains had MICs of chlorhexidine of 0.5–4 mg/L, except for one MRSA (MIC = 8 mg/L). All but four strains had MICs of miconazole of 1–4 mg/L (MIC = 16 mg/L, n = 3; MIC = 256 mg/L, n = 1). Miconazole/chlorhexidine (1:1 ratio) had a synergistic effect against 49/50 MRSA, 31/50 MSSA, 12/49 MRSP and 23/49 MSSP.ConclusionsSince the majority of these staphylococci, including methicillin-resistant isolates, had MICs that should be readily exceeded by topical skin application of these agents, their therapeutic efficacy for canine superficial pyoderma should be assessed. The synergistic interaction shown in vitro supports further clinical evaluation of miconazole/chlorhexidine combination therapy for staphylococcal infection.

Case–control risk factor study of methicillin-resistant Staphylococcus pseudintermedius (MRSP) infection in dogs and cats in Germany

Methicillin-resistant Staphylococcus pseudintermedius (MRSP) has emerged as a highly drug-resistant small animal veterinary pathogen. Although often isolated from outpatients in veterinary clinics, there is concern that MRSP follows a veterinary-hospital-associated epidemiology. This study's objective was to identify risk factors for MRSP infections in dogs and cats in Germany. Clinical isolates of MRSP cases (n=150) and methicillin-susceptible S. pseudintermedius (MSSP) controls (n=133) and their corresponding host signalment and medical data covering the six months prior to staphylococcal isolation were analysed by multivariable logistic regression. The identity of all MRSP isolates was confirmed through demonstration of S. intermedius-group specific nuc and mecA. In the final model, cats (compared to dogs, OR 18.5, 95% CI 1.8–188.0, P=0.01), animals that had been hospitalised (OR 104.4, 95% CI 21.3–511.6, P<0.001), or visited veterinary clinics more frequently (>10 visits OR 7.3, 95% CI 1.0–52.6, P=0.049) and those that had received topical ear medication (OR 5.1, 95% CI 1.8–14.9, P=0.003) or glucocorticoids (OR 22.5, 95% CI 7.0–72.6, P<0.001) were at higher risk of MRSP infection, whereas S. pseudintermedius isolates from ears were more likely to belong to the MSSP-group (OR 0.09, 95% CI 0.03–0.34, P<0.001). These results indicate an association of MRSP infection with veterinary clinic/hospital settings and possibly with chronic skin disease. There was an unexpected lack of association between MRSP and antimicrobial therapy; this requires further investigation but may indicate that MRSP is well adapted to canine skin with little need for selective pressure.

High diversity of Staphylococcus aureus and Staphylococcus pseudintermedius lineages and toxigenic traits in healthy pet-owning household members. Underestimating normal household contact?

Forty-three unrelated pet-owning households were screened in Spain to study the Staphylococcus aureus and Staphylococcus pseudintermedius nasal carriage, their genetic lineages and virulence traits. Sixty-seven healthy owners and 66 healthy pets were investigated. Isolates characterization was performed and potential interspecies transmission was assessed. S. aureus was present in 51.2% of households studied while S. pseudintermedius in 30.2%. Twenty-eight owners (41.8%) carried S. aureus: one methicillin-resistant S. aureus (MRSA) [t5173-ST8-SCCmecIVa] and 27 methicillin-susceptible S. aureus (MSSA). Three owners (4.5%) were colonized by methicillin-susceptible S. pseudintermedius (MSSP). Fifteen pets (22.7%) carried S. pseudintermedius: two methicillin-resistant S. pseudintermedius (MRSP) [ST71-SCCmecII/III; ST92-SCCmecV] and 13 MSSP; in addition, 8 pets (12.1%) presented MSSA. High diversity of spa and sequence types (STs) was detected. Typical livestock-associated S. aureus lineages (CC398, CC9) were observed in humans and/or companion animals and hospital and/or community-acquired S. aureus lineages (CC45, CC121, CC5, CC8) were detected among pets. Almost 40% of S. pseudintermedius were multidrug-resistant. S. aureus isolates harboured a remarkable high number of virulence genes. The expA gene was detected in 3 S. pseudintermedius isolates. Identical strains from both owners and their pets were identified in 5 households (11.6%): (a) four MSSA (t073-ST45/CC45, t159-ST121/CC121, t209-ST109/CC9, t021-ST1654([new])/singleton) and (b) one multidrug-resistant MSSP (ST142([new])). Highly clonally diverse and toxigenic S. aureus and S. pseudintermedius are common colonizers of healthy humans and pets. The presence of these bacterial species, virulence genes, and interspecies transmission detected, points out to consider pet ownership as a risk factor to acquire, maintain and spread, potential pathogenic bacteria.

Factors associated with methicillin-resistant versus methicillin-susceptible Staphylococcus pseudintermedius infection in dogs

To compare methicillin-resistant Staphylococcus pseudintermedius (MRSP) and methicillin-susceptible S pseudintermedius (MSSP) infections in dogs. Multicenter case-control study. Dogs with MRSP infections were matched, by hospital, with 2 MSSP controls, with the infections occurring immediately before and after the case infection. Signalment, historical, clinical, treatment, and outcome data were documented. Conditional logistic regression was performed. A manual stepwise backward elimination procedure was used to build the multivariable model. 56 case and 112 control dogs were enrolled. Pyoderma was the most common infection type in both groups. In the final multivariable model, systemic administration of antimicrobials within 30 days prior to infection was significantly associated with an MRSP versus an MSSP infection (OR, 9.9; 95% confidence interval, 3.59 to 27.53). The association of prior antimicrobial administration and MRSP infection indicated the potential impact of routine antimicrobial use in veterinary medicine on antimicrobial resistance and the need for prudent use of these important drugs. Mortality rate was not significantly different between MRSP and MSSP infections; the lack of a significant difference suggested that MRSP was inherently no more virulent than MSSP, provided the infection was properly diagnosed and appropriate treatment was started. Basic concepts such as prudent antimicrobial use and early diagnosis through timely submission of appropriate culture specimens therefore can be important measures to try to reduce the impact of this pathogen.

Carriage of Methicillin‐Resistant Staphylococcus pseudintermedius in Small Animal Veterinarians: Indirect Evidence of Zoonotic Transmission

Methicillin-resistant Staphylococcus pseudintermedius (MRSP) is increasingly reported in small animals and cases of human infections have already been described despite its recent emergence in veterinary practice. We investigated the prevalence of MRSP and methicillin-resistant Staphylococcus aureus (MRSA) among small animal dermatologists attending a national veterinary conference in Italy. Nasal swabs were obtained from 128 veterinarians, seven of which harboured MRSP (n = 5; 3.9%) or MRSA (n = 2; 1.6%). A follow-up study of two carriers revealed that MRSP persisted for at least 1 month in the nasal cavity. Methicillin-susceptible S. aureus (MSSA) was isolated from 32 (25%) conference participants, whereas methicillin-susceptible S. pseudintermedius (MSSP) was not detected, suggesting that MRSP may have a particular ability to colonize humans compared to MSSP. All isolates were characterized by spa typing. Methicillin-resistant isolates were further typed by antimicrobial susceptibility testing, SCCmec and multi-locus sequence typing. Two lineages previously associated with pets were identified among the five MRSP isolates; the European epidemic clone ST71-SCCmec II-III and ST106-SCCmec IV. One of the two MRSA isolates displayed a genotype (ST22- SCCmecIV) frequently reported in dogs and cats. MRSP isolates were resistant to more antimicrobial agents compared with MRSA isolates and displayed the typical multidrug resistance patterns of MRSP in pets. The 32 MSSA isolates belonged to 20 spa types and the most frequent types (t12, t15 and t166) were associated with common S. aureus lineages in humans (CC30 and CC45). Although low, the 3.9% MRSP carriage rate found among small animal dermatologists was surprising in consideration of the rare occurrence of S. pseudintermedius in humans, the lack of MSSP detection and the recent appearance of MRSP in Europe. As cases of human MRSP infection have been linked with pets, veterinarians should be aware of this zoonotic risk and proper preventative measures should be taken to avoid MRSP transmission from animal patients.

Identification of a predominant multilocus sequence type, pulsed-field gel electrophoresis cluster, and novel staphylococcal chromosomal cassette in clinical isolates of mecA-containing, methicillin-resistant Staphylococcus pseudintermedius

Methicillin resistance encoded by the mecA gene is increasingly observed in Staphylococcus pseudintermedius. Little is known about the population genetics of veterinary staphylococci bearing methicillin resistance. The aim of this study was to determine the relatedness of resistant bacteria and to compare them with methicillin-susceptible isolates. Multilocus sequence typing (MLST) and pulsed-field gel electrophoresis (PFGE) fragment profiling were performed on methicillin-resistant S. pseudintermedius (MRSP) and methicillin-susceptible S. pseudintermedius (MSSP) isolates obtained from canine samples submitted to the veterinary teaching hospital bacteriology service between 2006 and 2008. Multilocus sequence typing detected 20 different sequence types, 16 of which were not previously described. Methicillin-resistant isolates were predominantly ST 68, possessed the Staphylococcus aureus-associated staphylococcal chromosomal cassette mec (SCCmec) type VT and fell within the largest PFGE cluster; whereas methicillin-susceptible strains were more genetically diverse. This suggests that most methicillin resistance within the population of isolates tested originated from a single source which has persisted and expanded for several years.