Treatment of a lower urinary tract infection in a cat caused by a multi-drug methicillin-resistant Staphylococcus pseudintermedius and Enterococcus faecalis

Staphylococci and enterococci are common causes of urinary tract infections in cats. However, both species are rarely implicated together as causes of lower urinary tract infections associated with urethral obstruction. This report describes the first case of a multi-drug methicillin-resistant Staphylococcus pseudintermedius belonging to spa type t06 and Enterococcus faecalis urinary infection in a cat with pre-existing and recurrent urethral obstruction. Both species were isolated at >105CFU/ml from a cystocentesis urine specimen. Clinical and ultrasound features, results from urinalysis, urine culture, molecular typing and susceptibility testing by minimal inhibitory concentrations determination are described. Oral treatment with nitrofurantoin, the only antimicrobial agent that constituted a viable therapeutic option, had a positive outcome

Phenotypic and Molecular Traits of Staphylococcus coagulans Associated with Canine Skin Infections in Portugal

Funding Information: Funding: This work was supported by Project BIOSAFE funded by FEDER through the Programa Operacional Factores de Competitividade—COMPETE, by the Fundação para a Ciência e a Tecnologia (FCT, Portugal)—Grant LISBOA-01-0145-FEDER-030713, PTDC/CAL-EST/30713/2017 and by FCT through funds to GHTM (UID/04413/2020) and the CIISA Project (UID/CVT/00276/2020). Funding Information: This work was supported by Project BIOSAFE funded by FEDER through the Programa Operacional Factores de Competitividade?COMPETE, by the Funda??o para a Ci?ncia e a Tecnologia (FCT, Portugal)?Grant LISBOA-01-0145-FEDER-030713, PTDC/CAL-EST/30713/2017 and by FCT through funds to GHTM (UID/04413/2020) and the CIISA Project (UID/CVT/00276/2020). Publisher Copyright: © 2021 by the authors. Licensee MDPI, Basel, Switzerland.Staphylococcus coagulans is among the three most frequent pathogens of canine pyoderma. Yet, studies on this species are scarce. Twenty-seven S. coagulans and one S. schleiferi, corresponding to all pyoderma-related isolations from these two species at two veterinary laboratories in Lisbon, Portugal, between 1999 and 2018 (Lab 1) or 2018 (Lab 2), were analyzed. Isolates were identified by the analysis of the nuc gene and urease production. Antibiotic susceptibility towards 27 antibiotics was evaluated by disk diffusion. Fourteen antibiotic resistance genes were screened by PCR. Isolates were typed by SmaI-PFGE. Two S. coagulans isolates (2/27, 7.4%) were methicillin-resistant (MRSC, mecA+) and four (4/27, 14.8%) displayed a multidrug-resistant (MDR) phenotype. We observed resistance to penicillin (17/27, 63.0%), fluoroquinolones (11/27, 40.7%), erythromycin and clindamycin (3/27, 11.1%), fusidic acid (3/27, 11.1%) and tetracycline (1/27, 3.7%). The blaZ and erm(B) genes were carried by 16 and 1 isolates resistant to penicillin and erythromycin/clindamycin, respectively. Only three S. coagulans carried plasmids. The single S. schleiferi isolate presented an MDR phenotype. SmaI-PFGE revealed a limited genetic diversity of S. coagulans, with a predominant lineage present from 2001 to 2018. This study describes the first MRSC causing canine infection in Portugal and reveals a high burden of antimicrobial resistance, with the emergence of MDR phenotypes within the main lineages.publishersversionpublishe

Frequent biofilm production by Staphylococcus epidermidis causing infection in pets or colonizing veterinary staff

[P270]Aim: To characterize biofilm production by Staphylococcus epidermidis isolates causing infection in pets or colonizing veterinary staff. Methods: The study collection comprised 129 S. epidermidis isolates, from nasal colonization of vet-erinary staff (n = 112) and causing infection in cats and dogs (n = 17). The capacity to produce biofilm was evaluated by the Crystal Violet (CV) method, performed in flat-bottom 96-well polystyrene plates and biofilm production was categorized (as strong, moderate or weak) according to specific ranges of 570 nm optical density. The biofilm producer S. epidermidis RP62A was used as control in each assay. Biofilm-associated genes icaAD and aap were screened by PCR.Results: Amongst thehumancommensal isolates, the CV method enabled the identification of 66/112 (58.9 %) biofilm producers, of which 34/66 (51.5 %) were strong or moderate producers. In the group of pet infection isolates, 16/17 isolates (94.1 %) produced biofilm, of which 12/16 (75.0%) were strong or moderate producers. The majority of the isolates carried icaAD and/or aap genes. The genotype icaAD-aap- was associated with non-biofilm production whereas genotype icaAD+aap+ was linked to strong biofilm production. No correlation was found between any biofilm producing phenotype and the genotypes icaAD+aap-icaAD-aap+.Conclusion: These results evidence a high frequency of biofilm production by S. epidermidis either causing infection in pets or colonizing humans in close contact with them. The findings also highlight the importance of biofilm-associated genes other than ica in the biofilm phenotype.publishe

Prevalence of biofilm production in Staphylococcus epidermidis colonizing veterinary staff and students

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Characterization of plasmid encoded efflux determinants from Staphylococcus epidermidis

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Genetic diversity and antimicrobial resistance profiles of Staphylococcus pseudintermedius associated with skin and soft-tissue infections in companion animals in Lisbon, Portugal

Staphylococcus pseudintermedius is the main bacterial pathogen of skin and soft-tissue infections (SSTIs) in companion animals. Antimicrobial resistance in this species is a growing public health concern. This study aims to characterize a collection of S. pseudintermedius causing SSTIs in companion animals, establishing the main clonal lineages and antimicrobial resistance traits. The collection corresponded to all S. pseudintermedius (n = 155) causing SSTIs in companion animals (dogs, cats and one rabbit) collected between 2014 and 2018 at two laboratories in Lisbon, Portugal. Susceptibility patterns were established by disk diffusion for 28 antimicrobials (15 classes). For antimicrobials without clinical breakpoints available, a cut-off value (COWT) was estimated, based on the distribution of the zones of inhibition. The blaZ and mecA genes were screened for the entire collection. Other resistance genes (e.g., erm, tet, aadD, vga(C), dfrA(S1)) were searched only for those isolates showing an intermediate/resistance phenotype. For fluoroquinolone resistance, we determined the chromosomal mutations in the target genes grlA and gyrA. All the isolates were typed by PFGE following SmaI macrorestriction and isolates representative of each PFGE type were further typed by MLST. Forty-eight out of the 155  S. pseudintermedius isolates (31.0%) were methicillin-resistant (mecA+, MRSP). Multidrug-resistant (MDR) phenotypes were detected for 95.8% of the MRSP and 22.4% of the methicillin-susceptible (MSSP) isolates. Of particular concern, only 19 isolates (12.3%) were susceptible to all antimicrobials tested. In total, 43 different antimicrobial resistance profiles were detected, mostly associated with the carriage of blaZ, mecA, erm(B), aph3-IIIa, aacA-aphD, catpC221, tet(M) and dfr(G) genes. The 155 isolates were distributed within 129 PFGE clusters, grouped by MLST in 42 clonal lineages, 25 of which correspond to new sequence types (STs). While ST71 remains the most frequent S. pseudintermedius lineage, other lineages that have been replacing ST71 in other countries were detected, including ST258, described for the first time in Portugal. This study revealed a high frequency of MRSP and MDR profiles among S. pseudintermedius associated with SSTIs in companion animals in our setting. Additionally, several clonal lineages with different resistance profiles were described, evidencing the importance of a correct diagnosis and selection of the therapy

In vitro antimicrobial efficacy of two medical grade honey formulations against common high-risk meticillin-resistant staphylococci and Pseudomonas spp. pathogens

Background: Antimicrobial resistance is a problem in human and animal healthcare. Honey may be used for its wound healing properties and antimicrobial effects. Objective: To investigate the antimicrobial activity of two commercially available medical grade honeys (MGHs) against Staphylococcus spp. and Pseudomonas spp. isolates. Methods and materials: Two formulations, MGH1 (40% w/v honey) and MGH2 (80% w/v Manuka honey), were tested in vitro for minimal inhibitory concentrations (MIC) and minimal bactericidal concentrations (MBC) against 11 Staphylococcus and 11 Pseudomonas isolates at low [1.5 × 104 colony forming units (cfu)/well] and high (1.5 × 106 cfu/well) concentrations of inoculum, representing systemic and cutaneous bacterial loads during infection, respectively. Results: MGH2 showed a lower MIC against staphylococci than MGH1, although this was not statistically significant. MGH1 had stronger bactericidal effects against staphylococci than MGH2, although this effect was statistically significant only at the higher bacterial concentration (P < 0.01). For Pseudomonas spp., MGH1 had significantly higher antimicrobial activity (both MIC and MBC) than MGH2 against all isolates tested and at both bacterial concentrations (P < 0.05). Conclusions and clinical importance: Both MGHs were effective in vitro against common cutaneous pathogens including meticillin‐resistant staphylococci and Pseudomonas species. The higher efficacy of the MGH1 formulation against Pseudomonas and its consistent effects against staphylococci, while containing only half of the amount of honey compared to MGH2, invites further investigation of the mechanisms and clinical applications of MGH1

Biocide susceptibility in Staphylococcus epidermidis causing infection in pets

[P280]Aim: To characterize biocide susceptibility of Staphylococcus epidermidis isolates causing infection in pets and evaluate proposed biocide epidemiological cut-off (tentative ECOFF) values. Methods: The study comprised a collection of 17 S. epidermidis isolates collected from several infection sources in cats and dogs from 2001 to 2016. Susceptibility to benzalkonium chloride (BAC), cetrim-ide, chlorhexidine digluconate, tetraphenylphosphonium bromide (TPP), triclosan (TCL) and ethidium bromide (EtBr) was evaluated by microdilution MIC determination. The efflux pump genes qacA/B, qacG, qacJ and smr, associated with biocide resistance, and the triclosan resistance gene sh-fabI were screened by PCR.Results: The MIC distributions for all biocides and EtBr were analyzed against the tentative ECOFF val-ues proposed for these compounds (see other poster by Costa et al, ISSSI 2018). Non-wild-type (NWT) populations were detected towards BAC (6/17), TPP (8/17) and TCL (4/17) and EtBr (9/17). The NWT populations towards BAC, TPP and EtBr were associated with the presence of either qacA or smr plas-mid-encoded genes, whereas the TCL NWT population was linked to sh-fabI carriage. One isolate carry-ing the smr gene was only detected taking into consideration the proposed EtBr ECOFF.Conclusion: This study illustrates the frequently reduced susceptibility towards relevant biocides of S. epidermidis infecting pets. It also supports the tentative ECOFF values suggested for BAC, TPP and TCL, as criteria to detect S. epidermidis harboring biocide resistance genes. The establishment of such crite-ria would enable early detection of S. epidermidis isolates with a non-wild-type phenotype and improve therapeutics in veterinary medicine.publishe

Clonal lineages, antimicrobial resistance, and pvl carriage of staphylococcus aureus associated to skin and soft-tissue infections from ambulatory patients in Portugal

Staphylococcus aureus (S. aureus) is a leading cause of skin and soft-tissue infections (SSTIs) in the community. In this study, we characterized a collection of 34 S. aureus from SSTIs in ambulatory patients in Portugal and analyzed the presence of Panton–Valentine leucocidin (PVL)-encoding genes and antibiotic-resistance profile, which was correlated with genetic determinants, plasmid carriage, and clonal lineage. Nearly half of the isolates (15, 44.1%) were methicillin-resistant Staphylococcus aureus (MRSA) and/or multidrug resistant (MDR). We also detected resistance to penicillin (33/34, 97.1%), fluoroquinolones (17/34, 50.0%), macrolides and lincosamides (15/34, 44.1%), aminoglycosides (6/34, 17.6%), and fusidic acid (2/34, 5.9%), associated with several combinations of resistance determinants (blaZ, erm(A), erm(C), msr(A), mph(C), aacA-aphD, aadD, aph(3′ )-IIIa, fusC), or mutations in target genes (fusA, grlA/gyrA). The collection presented a high genetic diversity (Simpson’s index of 0.92) with prevalence of clonal lineages CC5, CC22, and CC8, which included the MRSA and also most MDR isolates (CC5 and CC22). PVL-encoding genes were found in seven isolates (20.6%), three methicillin-susceptible Staphylococcus aureus (MSSA) (ST152-agrI and ST30-agrIII), and four MRSA (ST8-agrI). Plasmid profiling revealed seventeen distinct plasmid profiles. This work highlights the high frequency of antimicrobial resistance and PVL carriage in SSTIs-related S. aureus outside of the hospital environment.publishersversionpublishe