In vitro antimicrobial activity of miconazole and polymyxin B against canine meticillin-resistant Staphylococcus aureus and meticillin-resistant Staphylococcus pseudintermedius isolates

Background – Meticillin‐resistant Staphylococcus aureus (MRSA) and meticillin‐resistant Staphylococcus pseudintermedius (MRSP) infections are increasingly reported in dogs, and these bacteria may be isolated from ear infections. Hypothesis/Objectives – The main aim of the present study was to investigate the in vitro antimicrobial activity of miconazole, polymyxin B and a combination of both against 24 canine MRSA and 50 canine MRSP isolates. The minimal inhibitory concentration (MIC) values of 12 other antimicrobial agents were also determined. Methods – All MIC values were determined according to a broth microdilution assay. Results – Acquired resistance was found to all tested agents, except for linezolid, miconazole and polymyxin B. The MIC values for miconazole and polymyxin B against MRSA were in the range of 4–8 and 8–64 μg/mL, respectively, while the MIC values for miconazole and polymyxin B against MRSP were in the range of 1–2 and 0.25–4 μg/mL, respectively. Using a combination of miconazole and polymyxin B, there was no evidence for enhanced in vitro activity of the combination (i.e. synergy) of both products. Nevertheless, MIC₉₀ values of the combination of these antimicrobial agents and of a commercial product containing both agents were at least 1000 times lower than the concentration present in the commercial product. Conclusions and clinical importance – These results indicate that the topical use of a combination of miconazole and polymyxin B in a 43.5:1 ratio may have potential for the treatment of MRSA‐mediated and MRSP‐associated otitis externa in dogs.     

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.     

大黄酸对伪中间葡萄球菌的抗菌活性及作用机制

伪中间葡萄球菌(Staphylococcus pseudintermedius)是犬脓皮病主要致病菌,具有多重耐药性,严重威胁犬及人类健康.本研究从脓皮病患犬中分离20株伪中间葡萄球菌,并揭示大黄酸对伪中间葡萄球菌的抗菌活性及抗菌机理.通过测定最小抑菌浓度、最低杀菌浓度、抑菌曲线和黏附抑制试验分析了大黄酸对伪中间葡萄球...     

In vitro miconazole susceptibility of meticillin-resistant Staphylococcus pseudintermedius and Staphylococcus aureus

Background – The emergence and dissemination of meticillin‐resistant staphylococci has created significant treatment challenges in veterinary medicine and increased interest in topical therapy for superficial infections. Concern has been expressed regarding the use of some topical antimicrobials in animals because of the potential for emergence of resistance, and additional options are required. Miconazole has limited antibacterial properties that include antistaphylococcal activity. Hypothesis/Objectives – The objective of this study was to assess the in vitro susceptibility of Staphylococcus pseudintermedius and Staphylococcus aureus to miconazole. Methods – In vitro susceptibility of 112 meticillin‐resistant S. pseudintermedius (MRSP), 53 meticillin‐resistant S. aureus (MRSA) and 37 meticillin‐susceptible S. pseudintermedius (MSSP) to miconazole was assessed using agar dilution. Results – The minimal inhibitory concentration (MIC) range, MIC₅₀ and MIC₉₀ for MRSP were 1–8, 2 and 4 μg/mL, respectively. Corresponding results for MRSA were 1–8, 2 and 6 μg/mL, and for MSSP 1–4, 2 and 2 μg/mL. The MIC for MSSP was a significantly lower MIC than that for both MRSP (P = 0.006) and MRSA (P < 0.001), while the MIC for MRSP was significantly lower than that for MRSA (P = 0.001). Conclusions and clinical importance – These in vitro data suggest that miconazole could be a useful therapeutic option for superficial infections caused by meticillin‐susceptible and meticillin‐resistant staphylococci, but proper clinical investigation is required.     

Antimicrobial activity of gallium against virulent Rhodococcus equiin vitro and in vivo

Rhodococcus equi, a facultative intracellular bacterium, causes severe pneumonia in foals. Evidence suggests that most foals become infected very early in life, when they have immature or ineffective innate immune responses. This study evaluated the antimicrobial activity of gallium against R. equi, as a potential chemoprophylactic and therapeutic agent. Rhodococcus equi was grown in media with various concentrations of gallium nitrate (GN), with and without excess iron. GN significantly inhibited growth and killed R. equi, and these effects were abolished with excess iron. Antimicrobial effects of Ga appear to be related to its interference with iron metabolism. Mice were treated orally with gallium maltolate (GaM), 10 or 50 mg/kg BW, or distilled H2O prior to and after experimental infection with R. equi. Six days post-infection, organs were harvested and R. equi concentrations assessed, and serum gallium concentrations determined. GaM was absorbed in a dose-dependent manner, and R. equi tissue burdens were greater in control mice than in all GaM-treated mice. GaM may aid in the control of disease by preventing development of overwhelming R. equi tissue burdens prior to the establishment of requisite innate and adaptive immune responses.     

Methicillin-resistant Staphylococcus aureus and Staphylococcus pseudintermedius in veterinary medicine

Staphylococci are important opportunistic pathogens in most animal species. Among the most relevant species are the coagulase positive species Staphylococcus aureus and Staphylococcus pseudintermedius. Methicillin resistance has emerged as an important problem in both of these organisms, with significant concerns about animal and public health. The relative importance of these staphylococci on different animal species varies, as do the concerns about zoonotic transmission, yet it is clear that both present a challenge to veterinary medicine.     

不同中药提取物对耐甲氧西林金黄色葡萄球菌的抑菌作用和逆转机制

采用(K-B)药敏纸片扩散法检测耐甲氧西林金黄色葡萄球菌(MRSA)的药物敏感性,并用二倍稀释法分别测定10种中药提取物对MRSA的最小抑菌浓度(MIC)和最小杀菌浓度(MBC),选取抑菌效果最好的中药提取物(黄芩)通过棋盘法测定其与5种常用抗生素的联合抑菌效果。观察黄芩不同亚抑菌浓度作用下对MRSA耐药性的逆转效果,并通过荧光实时定量PCR法检测耐药基因mec A在mRNA表达水平上的变化。结果表明,MRSA具有多重耐药性,10种中药提取物里黄芩的抑菌和杀菌效果最为显著,MIC和MBC均为156 mg/L,但其与5种抗生素没有显著的联合抗菌作用,在黄芩亚抑菌浓度130 mg/L作用20 h后,MRSA主要耐药基因mec A的表达量下调并恢复了对药物的相对敏感性。     

猪源耐甲氧西林金黄色葡萄球菌的耐药性及其SCCmec基因分型研究

为了解陕西猪场耐甲氧西林金黄色葡萄球菌(MRSA)的耐药表型及其SCCmec基因型分布情况,我们于2009年1月～7月在陕西地区6个规模化猪场,采集鼻腔拭子共计173份,利用选择性培养基分离金黄色葡萄球菌(SA),采用多重PCR方法扩增其中是否含有nuc基因和mecA基因进行MRSA鉴定;以琼脂稀释法进行药敏试验;并通过多重PCR方法对SCCmec进行基因分型。结果显示,调查的6个猪场中有2个猪场分离到MRSA,共分离到83株SA,其中有14株(16.9%)为MRSA,均为SCCmec-IVb型;药敏结果显示MRSA除对CFP、CHL、AMK和VAN敏感外,对CIP、OXA耐药率分别为57.1%和92.6%,对FOX、TET、ERY、甲氧嘧啶和卡那霉素的耐药率均为100%。结果显示了猪源SCCmec-IVb型MRSA株具有多重耐药特点,与人源SCCmec-IV型MRSA株耐药谱存在较大差异,这一结果提示猪源MRSA容易通过猪及其产品感染人。     

In vitro activity of fifteen antimicrobial agents against methicillin-resistant and methicillin-susceptible Staphylococcus intermedius

In this study the susceptibility of 91 methicillin-resistant and -susceptible Staphylococcus intermedius strains (MRSI and MSSI, respectively) against 15 antimicrobial agents was determined. The activity of the antimicrobial agents was studied at pH 7.2 and pH 8.5. Methicillin was more active at pH7.2 (28 strains methicillin-resistant) than at pH 8.5 (55 strains methicillin-resistant). Gentamicin showed excellent activity, with only 3 strains resistant at pH 8.5. However, gentamicin would have to be administered parenterally. Oxytetracy-cline cannot be recommended for treatment of canine staphylococcal dermatitis, due to the high percentage (over 25%) of strains that were found to be resistant. Clindamycin showed little activity in inhibiting growth of the strains studied, the percent resistance at pH 7.2 was 93.4%. Rifampin behaved differently at the two pH values. However, a close relationship was noted between methicillin-resistant and rifampin-resistant strains, particularly at the lower pH. Of the fluoroquinolones, ciprofloxacin or enrofloxacin would be a good useful alternative for the treatment of methicillin-resistant strains of S. intermedius . Lastly, very high resistance to sulphamethoxypyridazine was found, as was the case with trimethoprim and a combination of trimethoprim/ sulphamethoxypyridazine, against not only MRSI but also MSSI strains.     

In vitro antimicrobial activity of nitrofurantoin against Escherichia coli and Staphylococcus pseudintermedius isolated from dogs and cats

Minimum inhibitory concentrations (MIC) of nitrofurantoin were determined by agar dilution in 269 canine and feline isolates of Escherichia coli and Staphylococcus pseudintermedius, two of the most common bacterial species associated with urinary tract infection (UTI) in small animals. The MIC90 for E. coli and S. pseudintermedius were 32 and 16 μg/ml, respectively. All isolates, including multidrug-resistant strains of known genetic background, displayed MICs below the drug concentrations reported in canine urine following oral administration of nitrofurantoin. Preliminary data on mutant prevention concentration (MPC) and many years of nitrofurantoin usage in human medicine suggest that emergence of resistant mutants during treatment is not a critical issue for this drug. The study provides species-specific data on nitrofurantoin MIC distribution that can be used for setting dog- and cat-specific breakpoints. Although nitrofurantoin is not an appropriate first-line agent for empirical treatment of canine UTI due to toxicity and poor pharmacokinetic properties, it may be indicated for treatment of UTI caused by multidrug-resistant bacteria, which are otherwise difficult to treat using conventional veterinary antimicrobial agents.     

The selection in vitro of antibiotics with activity against intracellular S. aureus

The activity of a range of antibiotics on intracellular Staphylococcus aureus was examined using an in vitro system in which staphylococci survived within bovine neutrophils and extracellular organisms were killed by lysostaphin. Cloxacillin in the presence of lysostaphin caused a reduction in the number of viable intracellular S. aureus but cloxacillin alone failed to reduce such bacteria significantly. The cloxacillin appeared to sensitize the staphylococci to lysis by extracellular traces of lysostaphin following neutrophil disruption. Extracellular staphylococci which remained viable after exposure to cloxacillin in the absence of lysostaphin were subsequently killed more rapidly by neutrophils, but this was not found with bacteria exposed to cloxacillin while inside cells. Vancomycin with lysostaphin produced a similar but smaller sensitization effect but this antibiotic also appeared to increase survival of intracellular staphylococci when compared with controls, possibly by impeding neutrophil bactericidal mechanisms. The only other antibiotic to show significant intracellular killing was rifampicin, and in this case the action was independent of lysostaphin.     

Epidemiology of methicillin-resistant Staphylococcus aureus in dairy herds.

Sixty-eight methicillin-resistant Staphylococcus aureus strains were isolated from mastitis milk samples originating from 20 Belgian dairyherds. All these strains appeared to be representatives of one single strain which was probably of human origin. Evidence is presented indicating a rapid in vivo evolutionary change in this strain. The following characteristics were found to be variable: the production of beta haemolysin inversely connected with fibrinolysin (staphylokinase) activity; the production of lipase, enterotoxin B and delta haemolysin; the resistance to neomycin, chloramphenicol, tetracyclines, methicillin and spectinomycin associated with constitutive or inducible macrolide resistance.     

Efficacy of gallium maltolate against Lawsonia intracellularis infection in a rabbit model

Antimicrobial efficacy against Lawsonia intracellularis is difficult to evaluate in vitro, thus, the effects of gallium maltolate's (GaM) were investigated in a rabbit model for equine proliferative enteropathy (EPE). Juvenile (5–6‐week‐old) does were infected with 3.0 × 10⁸ L. intracellularis/rabbit and allocated into three groups (n = 8). One week postinfection, one group was treated with GaM, 50 mg/kg; one, with doxycycline, 5 mg/kg; and one with a sham‐treatment (control). Feces and blood were collected daily and weekly, respectively, to verify presence of L. intracellularis fecal shedding using qPCR, and seroconversion using immunoperoxidase monolayer assay. Rabbits were sacrificed after 1 week of treatment to collect intestinal tissues focusing on EPE‐affected sections. Intestinal lesions were confirmed via immunohistochemistry. No difference was noted between treatments regarding EPE‐lesions in jejunum (P = 0.51), ileum (P = 0.74), and cecum (P = 0.35), or in L. intracellularis fecal shedding (P = 0.64). GaM and doxycycline appear to have similar efficacy against EPE in infected rabbits.     

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。     

Antimicrobial activity of gallium nitrate against Mycobacterium avium subsp. paratuberculosis in neonatal calves

Background: Mycobacterium avium subsp. paratuberculosis (MAP), the agent of Johne's disease in cattle, is a facultative intracellular bacterium that is dependent on ferric iron for its survival and replication. Gallium (Ga), a trivalent semimetal that shares many similarities with ferric iron and functions as an iron mimic has been shown to have in vitro antimicrobial activity against several microorganisms, including MAP. Objectives: (1) To investigate the antimicrobial activity of Ga in calves experimentally infected with MAP; and (2) to monitor for potential adverse effects of Ga on calf health. Animals: Twelve Holstein calves. Methods: Randomized blind controlled experiment. Beginning at 10 days of age (study day 1), the experimental calves (n = 6) were treated with 20 mg/kg gallium nitrate daily for 45 days. On study days 4 and 5, all calves were challenged with a PO dose of a live field strain MAP. Treated calves were monitored daily for adverse effects. Calves were euthanized on study day 100, and 29 tissue samples and 1 fecal sample were collected from each calf. Samples were cultured for MAP by MGIT liquid culture system, Herrold's Egg Yolk Medium culture, or both. Results: No adverse effects were observed in the treated calves. Treatment was associated with a significant reduction in MAP tissue burden when compared with control calves (P = .017). Conclusions and Clinical Relevance: Chemoprophylactic treatment of calves with Ga before and during the period of high susceptibility decreased MAP tissue colonization in experimentally infected neonatal calves.     

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.     

Antibiotic treatments of a methicillin-resistant Staphylococcus pseudintermedius infection in a dog: a case presentation

We report the antibiotic treatments administered to a female dog with mastitis and successive pyoderma. Microbiological investigations allowed the identification of Staphylococcus pseudintermedius after 54 days of various antibiotic treatments. The isolate carried the mecA gene and was resistant to 9 of 15 tested antibiotics. Consistent antibiotic treatment of the infection was possible only after accurate microbiological diagnosis.