Meticillin-resistant strains of Staphylococcus pseudintermedius in companion animals

Sheila Crispin is chair of the new Advisory Council on the Welfare Issues of Dog Breeding. Here, she describes the background to the Council, outlines its priorities and offers some thoughts on the issues that need to be addressed.     

Antibiotic resistance and molecular characterization of ophthalmic Staphylococcus pseudintermedius isolates from dogs

The prevalence, virulence potential, and antibiotic resistance of ophthalmic Staphylococcus pseudintermedius (SP) isolated from dogs were examined. Sixty-seven Staphylococcus species were isolated from ophthalmic samples and surveyed for species-specific sequences in the Staphylococcus intermedius group (SIG) nuclease gene (SInuc), exfoliative toxin gene for SIG (siet), and antibiotic resistance genes (blaZ and mecA). PCR-restriction fragment length polymorphism analysis of the pta gene was also performed. Fifty isolates were identified as SIG strains, all of which were found to be SP. The blaZ gene was detected in 42 of the 50 SP strains and mecA gene was observed in 18 of the 50 SP strains. The 50 SP strains were most susceptible to amoxicillin/clavulanic acid (94%) and chlorampenicol (70%), and highly resistant to tetracycline (94%) and penicillin (92%). It was also found that 16 (88.9%) mecA-positive SP strains were resistant to oxacillin, tetracycline and penicillin. All mecA-positive SP were resistant to more than four of the eight tested antibiotics and therefore considered SP with multi-drug resistance (MDR). Our results indicate a high prevalence of antibiotic resistance genes in ophthalmic SP along with a close relationship between MDR SP strains and the mecA gene. Based on our findings, judicious administration of antibiotics to companion dogs is necessary.     

Antimicrobial susceptibility of Staphylococcus aureus and Staphylococcus pseudintermedius isolated from various animals

This study characterized the antimicrobial susceptibility of 221 Staphylococcus aureus isolated from various species, and 60 canine Staphylococcus pseudintermedius isolated from 1986 through 2000 at the Western College of Veterinary Medicine (WCVM). Resistance of S. aureus was most common to penicillin (31%) and tetracycline (14%); resistance of S. pseudintermedius to penicillin was present in 8% and to tetracycline in 34% of isolates. Resistance to trimethoprim/sulfamethoxazole was only seen among S. pseudintermedius, and there was no resistance to amoxicillin/clavulanate, ampicillin/sulbactam, cephalothin, amikacin, gentamicin, enrofloxacin, chloramphenicol, or rifampin among any isolate. Inducible clindamycin resistance was found in both S. aureus and S. pseudintermedius, highlighting the need for careful interpretation of culture and susceptibility test results. There were significant differences in the minimum inhibitory concentrations of penicillin, ciprofloxacin, enrofloxacin, clindamycin, erythromycin, chloramphenicol, and tetracycline between avian, bovine, equine, and porcine isolates.     

Case–control study of Staphylococcus lugdunensis infection isolates from small companion animals

Background – Coagulase‐negative Staphylococcus lugdunensis has recently been shown to cause invasive infections of people, which are similar in pathogenic effect to those caused by Staphylococcus aureus. Little is known about the pathogenicity of S. lugdunensis in companion animals. Objectives – To compare potential risk factors for infection, body sites affected, and whether cases and controls had been treated with antimicrobial drugs based upon susceptibility test results. Animals – Thirty‐three cases of S. lugdunensis infection (25 dogs, six cats and two small mammals) were identified between January 2003 and August 2011. Two Staphylococcus pseudintermedius controls, which were identified by the microbiology laboratory immediately before and after each S. lugdunensis case, were host‐species matched to each case. Methods – A retrospective case–control analysis. Results – During the period evaluated, the prevalence of S. lugdunensis infection was 1.3 cases per 10,000 hospital admissions for dogs and 0.95 cases per 10,000 admissions for cats (P = 0.453). In univariate analyses, S. pseudintermedius isolation was significantly associated with skin infections (P < 0.0001), while S. lugdunensis isolation was associated with the respiratory tract (P = 0.03) and other deep tissues (P = 0.005). Cases were less likely than controls to have been treated based upon susceptibility test results (P = 0.02). A conditional logistic regression analysis showed isolation of S. lugdunensis to be associated with recent (≤ 30 days) steroid administration (odds ratio, 17.72; 95% confidence interval, 2.35–132.82; P = 0.005); and in‐patient status (odds ratio, 9.67; 95% confidence interval, 2.18–42.88; P = 0.003). Conclusions – These results suggest that S. lugdunensis may cause invasive infections in companion animals, which should be treated with antimicrobials based upon susceptibility tests when available.     

Heterogeneity of Staphylococcus pseudintermedius isolates from atopic and healthy dogs

Staphylococcus pseudintermedius is part of the normal canine flora but frequently causes pyoderma in canine atopic dermatitis (AD). This study aimed to determine whether particular S. pseudintermedius strains were associated with AD and/or pyoderma. Ninety‐six S. pseudintermedius isolates from the ear, nares, perineum and lesions of 21 atopic and 16 healthy dogs were lysed with proteinase K and digested with 40 U SmaI. Restriction products were separated using pulsed‐field gel electrophoresis (PFGE) with an Oxford S. aureus control and lambda‐ladder DNA concatomer markers. A dendrogram was constructed by the unweighted pair group method. All isolates showed a ≥56% similarity coefficient. Nine distinct PFGE clusters were identified, as follows: five from both atopic and healthy dogs; three from atopic dogs only; and one from healthy dogs only. Nine clusters were isolated from the nares, eight from the perineum, five from the ears and six from pyoderma lesions. There were no significant differences in the frequency of isolation from atopic or healthy skin, body sites or infected lesions for any of the clusters. Two of six healthy dogs and 18 of 20 atopic dogs with multiple isolates had closely related isolates (less than three band differences) at more than one sampling site. Isolates from pyoderma lesions were closely related to at least one mucosal isolate in 11 of 16 dogs. Staphylococcus pseudintermedius isolates appear to be heterogeneous, and colonization or infection of atopic skin was not associated with any particular strain or cluster of strains.     

Prevalence and characterization of methicillin-resistant Staphylococcus pseudintermedius from symptomatic companion animals in Northern Italy: Clonal diversity and novel sequence types

The aim of this study was to assess the prevalence, the genotypic diversity, the antimicrobial resistance traits of canine and feline clinical methicillin-resistant Staphylococcus pseudintermedius (MRSP) isolates in a diagnostic laboratory in Italy during 2015-2016.All isolates were characterized by multilocus sequence typing (MLST), staphylococcal cassette chromosome (SCC)-mec typing and staphylococcal protein A (spa)-typing. The resistance profiles were assessed by antimicrobial susceptibility testing and confirmed genotypically by the detection of mecA gene and by microarray analyses.The prevalence of MRSP isolates was high (31.6%). All the strains were multidrug resistant and the most frequent clone was ST71-SCCmec type II-III. These results confirm a high prevalence of MRSP amongst clinical samples from pets in Italy. These isolates show multidrug resistance features that are of concern both in veterinary and human medicine for clinical and epidemiological reasons.     

Epidemiology of methicillin resistant Staphylococcus pseudintermedius in guide dogs in Finland

Background

Methicillin resistant Staphylococcus pseudintermedius (MRSP) and Staphylococcus aureus (MRSA) are common multi-drug resistant (MDR) bacteria in dogs. In 2012–2013 three dogs of the Guide Dog School of the Finnish Federation of the Visually Impaired were found to be MRSP positive. Guide dogs have regular contact with each other during their first year of life and prolonged contact when in training. Since dogs are placed in different parts of Finland after training, there is a risk for national spread of MDR bacteria. In this study the prevalence of MRSP and MRSA, as well as the risk factors for MRSP were determined in the Finnish guide dog population. MRSP isolates were investigated using molecular methods and compared to the earlier isolates.

Results

Out of 132 tested dogs 4 were MRSP positive thus giving the prevalence estimate of 3% (95% CI: 1–8%) for MRSP in the target population. MRSA was not detected (prevalence estimate 0%, 95% CI: 0–3%). Risk factors associated with MRSP were being a breeding bitch (OR = 8.4; 95% CI: 1.1–64.1, P = 0.012), the number of veterinary visits (OR = 1.23; 95% CI: 1.0–1.5, P = 0.025) and number of antimicrobial courses (OR = 1.63; 95% CI: 1.0–2.55; P = 0.035). Identified MRSP isolates belonged to five different sequence types (ST45, 71, 402, 403 and 404). All ST71 isolates carried SCCmec II-III, while the SCCmec type of the ST45 and ST402 (a single locus variant of ST45) isolates were non-typeable with the method used.

Conclusions

MRSP and MRSA had low prevalence in the studied dog population despite the close contact between dogs, and the MRSP population was heterogenic. Antimicrobial therapy and veterinary visits are risk factors for MRSP even among a small case group.

Electronic supplementary material

The online version of this article (doi:10.1186/s13028-015-0129-8) contains supplementary material, which is available to authorized users.     

Comparative mutant prevention concentration and mechanism of resistance to veterinary fluoroquinolones in Staphylococcus pseudintermedius

Background – The problem of antibacterial drug resistance is increasing worldwide, in part due to the therapeutic concentrations currently used based on the minimal inhibitory concentration (MIC) as a measure of potency are often the very concentrations required to selectively enrich the resistant mutant portion of the population. A mutant prevention concentration (MPC)‐based dosing strategy is suggested to improve the therapeutic outcome based on the MIC. Objective – Our aim was to investigate the MPC and mechanism of resistance to various fluoroquinolones using recent Staphylococcus pseudintermedius isolates from canine pyoderma. Methods – The broth microdilution method for MIC and a series of agar plates containing different concentrations of fluoroquinolones were inoculated with ～10¹⁰ colony‐forming units of the bacterial culture for MPC were used. PCR was used to identify mutation in the resistant isolates. Results – The rank order of potency based on MIC and MPC was ciprofloxacin = enrofloxacin ≥ marbofloxacin > difloxacin ≥ orbifloxacin. Integrating our data with reported pharmacokinetic data at the recommended dose ranges revealed that only high doses of ciprofloxacin, enrofloxacin and marbofloxacin could achieve a maximal plasma concentration (C_max) greater than the MPC of 90% of isolates (C_max/MPC₉₀). The overall rank of potency against S. pseudintermedius, based on C_max/MIC, C_max/MPC, the area under concentration–time curve (AUC)/MIC and AUC/MPC values, was in decreasing order: enrofloxacin > ciprofloxacin ≥ marbofloxacin ≥ orbifloxacin = difloxacin. Sequencing of the quinolone resistant determining region of gyrA, gyrB, grlA and grlB of resistant strains showed a base‐pair substitution in both gyrA and gyrB that resulted in Ser‐84 to Leu and Ser‐80 to Arg amino acid changes, respectively. Conclusions and clinical importance – High doses of ciprofloxacin, enrofloxacin and marbofloxacin could minimize the selection of resistant mutants, whereas the possibility of selecting mutants with the conventional doses of difloxacin and orbifloxacin, and low clinical doses of all fluoroquinolones, seems high.     

广东省宠物源伪中间型葡萄球菌的耐药性调查研究

为调查广东省宠物源伪中间型葡萄球菌(Staphylococcus pseudintermedius)的携带和耐药情况,从小动物临床采集皮肤、耳部和鼻腔等部位的样品,分离鉴定伪中间型葡萄球菌,并采用琼脂稀释法和纸片扩散法对伪中间型葡萄球菌进行抗菌药物敏感性检测,采用PCR方法检测mecA基因。结果表明,从898个样品中共分离到144株伪中间型葡萄球菌,检出率为16.0%,皮肤、耳部和鼻腔的伪中间型葡萄球菌检出率分别为20%(64/319)、17.8%(64/359)和7.5%(3/40)。抗菌药物敏感性检测和mecA基因检测结果显示有89株(61.8%)为耐甲氧西林伪中间型葡萄球菌(MRSP)。伪中间型葡萄球菌对氨苄西林、红霉素、阿奇霉素、克林霉素和四环素的耐药率均在80%以上,对利福平和阿米卡星的耐药率低于8%,对万古霉素、利奈唑胺、喹奴普汀/达福普汀和替考拉宁均敏感。大部分(>69%)MRSP对红霉素、克林霉素、泰乐菌素、阿奇霉素、复方新诺明、四环素、环丙沙星和恩诺沙星等抗菌药物表现耐药。本研究结果表明宠物源伪中间型葡萄球菌的耐药情况严重且MRSP的检出率高,应高度重视宠物源细菌耐药问题。     

Prevalence of canine methicillin resistant Staphylococcus pseudintermedius in a veterinary diagnostic laboratory in Italy

The overall prevalence of methicillin-resistant Staphylococcus pseudintermedius (MRSP) was 2% (10/590) among 590 canine specimens submitted to an Italian veterinary diagnostic laboratory during a two-month period, and 21% (10/48) among Staphylococcus intermedius group (SIG) isolates. All methicillin-resistant strains exhibited additional resistance to fluoroquinolones, gentamicin, lincosamides, tetracyclines, and potentiated sulfonamides, belonged predominantly to spa type t02 and harboured SCCmec type II–III cassette.     

山羊源伪中间葡萄球菌的分离鉴定及耐药性检测

伪中间葡萄球菌是一种常见的致病菌,多引起宠物不同程度感染。2016年3月初,吉林省某羊场育肥羊出现零星死亡,病羊主要表现为呼吸系统症状。采集病羊呼吸系统各器官,以进行细菌的分离、培养。分离菌株经培养特性、染色镜检、生化试验及16S r DNA序列分析,确定为伪中间葡萄球菌,其对小鼠有致病性。对分离到的菌株通过微量稀释法进行药物敏感性测定,结果显示,对庆大霉素、链霉素、红霉素、阿莫西林、头孢氨苄、磺胺间甲氧嘧啶、恩诺沙星、强力霉素耐药;对阿米卡星、氟苯尼考、环丙沙星敏感。     

Prevalence of canine methicillin resistant Staphylococcus pseudintermedius in a veterinary diagnostic laboratory in Italy

The overall prevalence of methicillin-resistant Staphylococcus pseudintermedius (MRSP) was 2% (10/590) among 590 canine specimens submitted to an Italian veterinary diagnostic laboratory during a two-month period, and 21% (10/48) among Staphylococcus intermedius group (SIG) isolates. All methicillin-resistant strains exhibited additional resistance to fluoroquinolones, gentamicin, lincosamides, tetracyclines, and potentiated sulfonamides, belonged predominantly to spa type t02 and harboured SCCmec type II–III cassette.     

Screening of Staphylococcus aureus, Staphylococcus intermedius, and Staphylococcus schleiferi isolates obtained from small companion animals for antimicrobial resistance: a retrospective review of 749 isolates (2003-04)

Companion animal staphylococcal isolate antibiograms were screened retrospectively to determine the frequency of methicillin-resistant (MR) infection by Staphylococcus aureus, Staphylococcus intermedius, and Staphylococcus schleiferi. Rates of MR were: S. aureus 35%, S. intermedius 17%, and S. schleiferi 40%. Frequency of isolation of methicillin-resistant S. aureus (MRSA) from dogs and cats was similar, whereas methicillin-resistant S. intermedius (MRSI) and methicillin-resistant S. schleiferi (MRSS) were significantly more common in dogs. MRSS was more commonly associated with superficial (skin and ear canal) infections, whereas MRSA was more commonly associated with deep infections. The MR strain resistance pattern to other classes of antibiotics was also investigated. MRSA was resistant to the most classes of antibiotics, followed by MRSI, while MRSS maintained the most favourable susceptibility profile. MR staphylococci may pose a significant risk to animal and public health. Therefore, to avoid selecting for resistant strains in cases of suspected staphylococcal infection, clinicians should consider culture and susceptibility testing early in the course of treatment.     

Antimicrobial resistance of Staphylococcus pseudintermedius isolates from healthy dogs and dogs affected with pyoderma in Japan

Staphylococcus pseudintermedius strains were isolated from healthy dogs and dogs with pyoderma in 2000-2002 and 2009. All the isolates from dogs with pyoderma in 1999-2000 and from healthy dogs in 2000-2002 and 2009 were susceptible to cefalexin and/or other cephalosporins and oxacillin. However, 7.1-12.5 and 11.4% of S. pseudintermedius isolates from dogs with pyoderma in 2009 were resistant to cephalosporins and oxacillin, respectively. All S. pseudintermedius isolates from dogs with pyoderma in 1999-2000 and those from healthy dogs in 2000-2002 were susceptible to fluoroquinolones; however, 50% of the S. pseudintermedius strains isolated from dogs with pyoderma in 2009 and 30% of the S. pseudintermedius strains isolated from healthy dogs in 2009 were resistant to fluoroquinolones. Of the 21 oxacillin-resistant S. pseudintermedius (MRSP) isolates, 11 carried SCCmec type V and 10 carried hybrid SCCmec types II-III. Staphylococcus pseudintermedius strains that were resistant to only one of three fluoroquinolones had a mutation in the quinolone resistance determination region of grlA, whereas S. pseudintermedius strains that were resistant to two or more fluoroquinolones had mutations in the quinolone resistance determination regions of both grlA and gyrA.