RESUMO
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.
Assuntos
Biofilmes , Infecções Estafilocócicas , Animais , Titânio , Matriz Extracelular de Substâncias Poliméricas , Staphylococcus/genéticaRESUMO
AIMS: The purpose of this study was to characterize the capacity for biofilm formation, antimicrobial resistance rates, and search for genetic determinants of resistance and virulence in the species. METHODS AND RESULTS: Strains were collected from asymptomatic and infected dogs. Identification was conducted using matrix-assisted laser desorption ionization-time of flight (MALDI-TOF), antimicrobial susceptibility using disk diffusion and PCR targeting mecA. Biofilm formation was evaluated on a microtiter plate assay. A total of 27 strains were selected for whole-genome sequencing. We identified 111 Staphylococcus coagulans. The highest number was obtained from infected dogs. The highest resistance rates were observed for penicillin, gentamicin, and ciprofloxacin/erythromycin. Twelve strains were characterized as resistant to methicillin. All isolates had the ability to form biofilm and were strong producers. Among Methicillin Resistant Staphylococcus coagulans (MRSC), SCCmec types IIIA, and Vc were identified. Acquired resistance genes, such as aac(6')-aph(2''), tet(K), blaZ, qacG, qacJ, and erm(C) were found. Different virulence genes were identified. Of note, Panton-Valentine Leucocidin was highly prevalent among the isolates. CONCLUSION: Staphylococcus coagulans had a high isolation rate among infected dogs and demonstrated significant resistance to commonly used antibiotics such as penicillin and gentamicin.
Assuntos
Staphylococcus aureus Resistente à Meticilina , Infecções Estafilocócicas , Cães , Animais , Staphylococcus aureus Resistente à Meticilina/genética , Leucocidinas/genética , Prevalência , Infecções Estafilocócicas/epidemiologia , Infecções Estafilocócicas/veterinária , Testes de Sensibilidade Microbiana , Brasil/epidemiologia , Antibacterianos/farmacologia , Staphylococcus/genética , Meticilina , Gentamicinas/farmacologiaRESUMO
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.
Assuntos
Staphylococcus aureus Resistente à Meticilina , Otite , Pioderma , Infecções Estafilocócicas , Cães , Animais , Humanos , Resistência a Meticilina , Tipagem de Sequências Multilocus/veterinária , Filogenia , Infecções Estafilocócicas/epidemiologia , Infecções Estafilocócicas/veterinária , Infecções Estafilocócicas/microbiologia , Antibacterianos/farmacologia , Testes de Sensibilidade MicrobianaRESUMO
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.