Antimicrobial susceptibility of Pseudomonas aeruginosa from dogs and cats as well as Arcanobacterium pyogenes from cattle and swine as determined in the BfT-GermVet monitoring program 2004-2006

During the BfT-GermVet monitoring program, Pseudomonas (P) aeruginosa from dogs and cats (n = 99) as well as Arcanobacterium (A.) pyogenes from cattle and swine (n = 90) were examined for their antimicrobial susceptibility. In general, P. aeruginosa is known to be resistant against many antimicrobial agents whereas A. pyogenes is thought to be susceptible to most agents in-vitro. However, representative and actual minimum inhibitory concentration (MIC) values are missing for both veterinary pathogens. In the present study, MIC values were determined and categorized according to the recommendations given in the Clinical and Laboratory Standards Institute (CLSI) documents M31-A2 and M31-S1. For susceptibility testing of A. pyogenes, the CLSI methodology was slightly modified. Specific breakpoints were not available for most of the antimicrobial agents tested. P. aeruginosa isolates from infections of the skin, ear and mouth as well as the urinary and genital tract of dogs and cats were either resistant or exhibited high MIC values to most antimicrobial agents tested. However, gentamicin resistant isolates were observed in only 27% and 11% (intermediate isolates 29% and 39%), respectively. For the same bacterium/host animal/organ system combinations, enrofloxacin resistance was detected in only 24% and 11% of the isolates (intermediate isolates 49% and 61%). For A. pyogenes, resistance was most prevalent against tetracycline (33%-56%, bovine and porcine isolates) and sulfonamides (26%-40%, bovine isolates).     

Susceptibility of bacterial pathogens against lincomycin/spectinomycin (1/2), penicillin G/neomycin (1/1), and penicillin G/dihydrostreptomycin (1/1) as determined in the BfT-GermVet monitoring program 2004-2006

A total of 368 bacterial pathogens, including 72 coagulase-positive and coagulase-variable staphylococci, 97 beta-haemolytic streptococci, 51 Escherichia coli, 75 Pasteurella multocida, 25 Mannheimia haemolytica, 25 Pseudomonas aeruginosa, and 23 Arcanobacterium pyogenes, were investigated for their susceptibility to the three combinations of antimicrobial agents lincomycin/spectinomycin (1/2), penicillin G/neomycin (1/1), and penicillin G/dihydrostreptomycin (1/1) in comparison to their susceptibility to the corresponding single substances. When comparing the minimum inhibitory concentrations (MICs) determined for any of the three combinations with those for the single substances, the lowest MIC of one of the two substances usually determined the MIC of the combination.This observation was made for all three combinations and all bacterial pathogens tested.Thus, it is assumed that the combination of lincomycin with spectinomycin as well as that of penicillin with either neomycin or dihydrostreptomycin resulted in an extended spectrum of target bacterial pathogens rather than in an increase in antimicrobial efficacy.     

国家动物源细菌耐药性监测工作的探讨和建议

介绍了国外动物源细菌耐药性监测工作的情况及目标,对我国的动物源细菌耐药性监测工作需考虑的许多因素进行了介绍和讨论,包括动物品种、食品、采样方法、样本的收集、细菌品种、抗菌药物、标准药敏试验法、质控、结果记录和数据库设计、报告和结果分析等,并对今后的动物源细菌耐药性监测工作提出了建议。     

Layout proposal for a microtitre plate to be used in routine antimicrobial susceptibility testing of bacteria from infections of dogs and cats

The determination of minimum inhibitory concentrations by broth microdilution is recommended as method of choice for susceptibility testing of veterinary bacterial pathogens. Accordingly, broth microdilution is used in veterinary routine diagnostic laboratories at a progressive rate. To reduce the costs of susceptibility testing, it is reasonable to develop widely accepted uniform microtitre plate layouts that are produced in large quantities. Such microtitre plate layouts have already been developed and published for the susceptibility testing of pathogens from food-producing animals. However, a microtitre plate layout, especially designed for the testing of bacteria from dogs and cats, should be available, too. The choice of the antimicrobial agents or combinations of antimicrobial agents to be included in a suitable layout should be based on the following criteria: (1) the approval and availability of an antimicrobial agent or combination of agents, (2) known cross-resistances, and (3) availability of approved clinical breakpoints. The latter point is of particular importance for the choice of the numbers of concentrations per antimicrobial agent tested and the range of test concentrations. Taking into account these aspects, a science-based layout proposal for microtitre plates, which are suitable for routine testing of bacteria from dogs and cats, is presented and discussed.     

Antimicrobial susceptibility pattern of Helicobacter suis strains

Helicobacter suis is a very fastidious porcine gastric pathogen, which is also considered to be of zoonotic importance. In vitro antimicrobial susceptibility cannot be determined using standard assays, as this agent only grows in a biphasic medium with an acidic pH. Therefore, a combined agar and broth dilution method was used to analyse the activity of nine antimicrobial agents against nine H. suis isolates. After 48 h microaerobic incubation, minimal inhibitory concentrations (MICs) were determined by software-assisted calculation of bacterial growth. Only for enrofloxacin a bimodal distribution of MICs was demonstrated, indicating acquired resistance in one strain, which showed an AGT→AGG (Ser→Arg) substitution at codon 99 of gyrA. In conclusion, the assay developed here is suitable for determination of the antimicrobial susceptibility of H. suis isolates, although activity of acid sensitive antimicrobial agents may be higher than predicted from MIC endpoints.     

OIE抗菌药耐药性国际标准

兽用抗菌药耐药性已经成为一个全球普遍关注的公共健康问题,各国际组织都积极采取相应的措施控制耐药性的产生和蔓延。介绍了国际组织世界动物卫生组织OIE制定的五个国际标准,包括协调抗菌药耐药性监督和检测程序指南、畜牧业抗菌药消耗量监测指南、兽用抗菌药慎用指南、抗菌药敏感性检测的实验室方法指南、动物源抗菌药耐药性对公共健康潜在影响的风险分析方法指南,以期为我国政策制定者和决策者参照国际标准制定出符合我国国情的耐药性相关指南。     

抗菌肽对抗生素耐药菌株的抑菌活性研究

[目的]研究抗菌肽对抗生素耐药菌株的抑菌活性。[方法]利用抗性平板划线法从腹泻病牛血便中筛选分离出1株耐药菌,通过16S rDNA序列进行鉴定,采用琼脂孔穴扩散法通过梯度盐酸壮观霉素（spectinomycin,Spe⁺）、氨苄青霉素（ampicillin,Amp⁺）、硫酸卡那霉素（kanamycin,Kan⁺）和氯霉素（chloramphenicol,Cm⁺）试验确定该菌药敏特性,并利用1种抗菌肽制剂对该菌株进行药敏试验。[结果]经BLAST比对分析该菌16S rDNA序列,鉴定该耐药菌为科氏葡萄球菌（Staphylococcus cohnii）,此菌对Amp⁺敏感,但对试验中其他抗生素均有耐药性,各梯度抗菌肽对该耐药菌均具有明显的抑菌活性。[结论]抗菌肽能有效抑制耐药科氏葡萄球菌的生长,有望在畜牧生产中代替抗生素使用。     

Antimicrobial resistance of Actinobacillus pleuropneumoniae isolated from swine

The aim of this retrospective study was to evaluate the antimicrobial resistance rates and the trend in resistance of Actinobacillus pleuropneumoniae isolated from pigs in Italy from 1994 to 2009. A total of 992 A. pleuropneumoniae isolates were tested for their susceptibility to a panel of antimicrobial agents in a disk diffusion method. Resistance to 7 drugs (amoxicillin, amoxicillin/clavulanic acid, ampicillin, cefquinome, cotrimoxazole, penicillin G and tilmicosin) showed a significant increasing trend over the time, while for 2 drugs (gentamycin and marbofloxacin) a significant decrease was observed. Resistance to the remaining 14 antimicrobial agents tested did not change significantly over the study period. Most of the isolates retained high susceptibility to antimicrobials usually effective against A. pleuropneumoniae such as amphenicols, fluoroquinolones and ceftiofur. However, high rates of resistance were observed for potentiated sulfa drugs, tetracyclines and penicillins which are currently recommended antimicrobials for pig pleuropneumonia therapy. Our results suggest the importance of continued monitoring of A. pleuropneumoniae clinical isolates in order to choose the most appropriate treatment of infections and to control the increase of resistance to currently used antimicrobials.     

Antibiotic susceptibility of bacteria isolated from infections in cats and dogs throughout Europe (2002–2009)

A monitoring program of the pre-treatment susceptibility of clinical isolates of bacteria from diseased dogs and cats was active between the years 2002 and 2009. Susceptibility of each isolated strain to a panel of nine antibiotics (amoxicillin/clavulanic acid, ampicillin, penicillin, clindamycin, doxycycline, enrofloxacin, marbofloxacin, trimethoprim and trimethoprim/sulfamethoxazole) was assessed. The Minimum Inhibitory Concentration (MIC) of marbofloxacin was also determined by a standardized microdilution technique following CLSI recommendations. In total, 1857 bacterial strains were collected throughout Europe from cases of otitis, respiratory, urinary and dermatological infections. Although bacterial susceptibility varied for each of the antibiotics within the panel, patterns of susceptibility were similar to those described in the literature for comparable time periods and geographical areas. With a clinical resistance varying from 0 to 14.48% against the isolated strains, marbofloxacin susceptibility was very high and remains an effective antibiotic for the treatment of otitis, urinary, respiratory and dermatological infections in companion animals.     

Monitoring of antimicrobial resistance among food animals: principles and limitations

Large amounts of antimicrobial agents are in the production of food animals used for therapy and prophylactics of bacterial infections and in feed to promote growth. The use of antimicrobial agents causes problems in the therapy of infections through the selection for resistance among bacteria pathogenic for animals or humans. Current knowledge regarding the occurrence of antimicrobial resistance in food animals, the quantitative impact of the use of different antimicrobial agents on selection for resistance and the most appropriate treatment regimes to limit the development of resistance is incomplete. Programmes monitoring the occurrence and development of resistance are essential to determine the most important areas for intervention and to monitor the effects of interventions. When designing a monitoring programme it is important to decide on the purpose of the programme. Thus, there are major differences between programmes designed to detect changes in a national population, individual herds or groups of animals. In addition, programmes have to be designed differently according to whether the aim is to determine changes in resistance for all antimicrobial agents or only the antimicrobial agents considered most important in relation to treatment of humans. In 1995 a continuous surveillance for antimicrobial resistance among bacteria isolated from food animals was established in Denmark. Three categories of bacteria, indicator bacteria, zoonotic bacteria and animal pathogens are continuously isolated from broilers, cattle and pigs and tested for susceptibility to antimicrobial agents used for therapy and growth promotion by disc diffusion or minimal inhibitory concentration determinations. This programme will only detect changes on a national level. However, isolating the bacteria and testing for several antimicrobial agents will enable us to determine the effect of linkage of resistance. Since 1995 major differences in the consumption pattern of different antimicrobial agents have occurred in Denmark. The Danish monitoring programme has enabled us to determine the effect of these changes on the occurrence of resistance. The Danish monitoring is, however, not suited to determine changes on a herd level or to detect emergence of new types of resistance only occurring at a low level.     

Repurposing Ionophores as novel antimicrobial agents for the treatment of bovine mastitis caused by Gram‐positive pathogens

Increasing reports of multidrug‐resistant bacterial infections in animals has created a need for novel antimicrobial agents that do not promote cross‐resistance to critically important antimicrobial classes used in human medicine. In response to the recent emergence of antimicrobial resistance in several bovine mastitis pathogens, in vitro antimicrobial susceptibility was determined for four polyether ionophores (lasalocid, monensin, narasin and salinomycin) against Staphylococcus spp. and Streptococcus spp. isolated from clinical cases. In addition, erythrocyte haemolysis and WST ‐1 cell proliferation assays were used to assess in vitro mammalian cell cytotoxicity and biofilm susceptibility testing was performed using the minimum biofilm eradication concentration (MBEC ?) biofilm assay. Lasalocid, monensin, narasin and salinomycin exhibited bacteriostatic antimicrobial activity against all pathogens tested, including methicillin‐resistant staphylococci, with MIC ₉₀ values <16 μg/ml. Narasin and monensin displayed the least toxicity against mammalian cell lines and all compounds significantly reduced viable cell numbers in a Staphylococcus aureus biofilm. Based on in vitro characterization, all four ionophores offer potentially novel treatments against bovine mastitis but in vivo studies will be essential to determine whether acceptable safety and efficacy is present following intramammary administration.     

Antimicrobial susceptibility of Pasteurella multocida and Bordetella bronchiseptica from dogs and cats as determined in the BfT-GermVet monitoring program 2004-2006

A total of 92 canine/feline Pasteurella multocida strains form respiratory tract infections or infections of skin/ear/mouth as well as 42 canine/feline Bordetella bronchiseptica strains from respiratory tract infections were investigated for their susceptibility to antimicrobial agents. While the P. multocida strains were susceptible to all antimicrobial agents tested - except sulfonamides -, a considerable number of the B. bronchiseptica strains was resistant or exhibited high MIC values against a number of antimicrobial agents including penicillin G, oxacillin, cefazolin, ceftiofur, cefquinome, sulfamethoxazole, and trimethoprim/sulfamethoxazole.     

Antimicrobial susceptibility of Escherichia coli from swine, horses, dogs and cats as determined in the BfT-GermVet monitoring program 2004-2006

A total of 417 isolates of Escherichia coli collected from five animal species/organ system combinations from swine [urinary/genital tract (UGT) incl. mastitis metritis agalactia syndrome], horses [genital tract (GT)] and dogs/cats [respiratory tract (RT), UGT and gastrointestinal tract (GIT)] were analysed quantitatively for their susceptibility against different antimicrobial agents by determination of minimum inhibitory concentrations. Regardless of which animal species the strains originated from, resistance appeared most frequently against sulfamethoxazole (18-59%), tetracycline (14-54 %), and ampicillin (14-39%). High percentages of intermediate isolates were observed for cephalothin (39-46 %). In general, low prevalences of resistance were detected for amoxicillin/clavulanic acid (1-4%), gentamicin (1-9%), and cefazolin (0-11%). Generally speaking, the antimicrobial resistance situation among E. coli isolates from horses and small animals is relatively good.     

Survey of antimicrobial susceptibility testing practices of veterinary diagnostic laboratories in the United States

OBJECTIVE: To describe antimicrobial susceptibility testing practices of veterinary diagnostic laboratories in the United States and evaluate the feasibility of collating this information for the purpose of monitoring antimicrobial resistance in bacterial isolates from animals. DESIGN: Cross-sectional study. PROCEDURES: A questionnaire was mailed to veterinary diagnostic laboratories throughout the United States to identify those laboratories that conduct susceptibility testing. Nonrespondent laboratories were followed up through telephone contact and additional mailings. Data were gathered regarding methods of susceptibility testing, standardization of methods, data management, and types of isolates tested. RESULTS: Eighty-six of 113 (76%) laboratories responded to the survey, and 64 of the 86 (74%) routinely performed susceptibility testing on bacterial isolates from animals. Thirty-four of the 36 (94%) laboratories accredited by the American Association of Veterinary Laboratory Diagnosticians responded to the survey. Laboratories reported testing > 160,000 bacterial isolates/y. Fifty-one (88%) laboratories reported using the Kirby-Bauer disk diffusion test to evaluate antimicrobial susceptibility; this accounted for 65% of the isolates tested. Most (87%) laboratories used the NCCLS (National Committee for Clinical Laboratory Standards) documents for test interpretation. Seventy-five percent of the laboratories performed susceptibility testing on bacterial isolates only when they were potential pathogens. CONCLUSIONS: The veterinary diagnostic laboratories represent a comprehensive source of data that is not easily accessible in the United States. Variability in testing methods and data storage would present challenges for data aggregation, summary, and interpretation.     

Antimicrobial susceptibility testing for bovine respiratory disease: Getting more from diagnostic results

Bovine respiratory disease (BRD) is one of the most common diseases of cattle worldwide. Given the significant bacterial component of this disease, antimicrobial agents remain one of the mainstays of therapy. However, the potential welfare and economic impact resulting from the selection of inappropriate antimicrobial therapy for BRD poses significant risks to both animal and animal owner. To determine the ‘best’ antimicrobial agent for a specific case, the decision-making process needs to incorporate all available evidence, often including the results of bacterial culture and antimicrobial susceptibility testing. While antimicrobial susceptibility testing can be a valuable diagnostic tool, integrating the test results into the clinical decision making process can be a challenging experience. This review details the process by which interpretive criteria for susceptibility tests are developed. Principles for how to best integrate antimicrobial susceptibility testing, both at the individual animal test and aggregate test levels, into the clinical decision making process are discussed. Non-traditional testing methodologies and how they may improve susceptibility testing in the future are also reviewed.     

Evaluation of the antimicrobial activity of florfenicol against bacteria isolated from bovine and porcine respiratory disease

Antimicrobial susceptibility of florfenicol (FFC) against 243 bacterial agents isolated in Korea from cattle and pigs with respiratory disease were investigated by agar diffusion and microdilution broth methods following the recommendations provided by the National Committee for Clinical Laboratory Standards. All Actinobacillus pleuropnemoniae, Pasteurella multocida, Mannheimia haemolytica and 98.6% of the Bordetella bronchiseptica isolates were susceptible to FFC, which as significantly more effective than the other antibiotics used in this study. FFC also showed high in vitro antimicrobial activities (MIC(90) < or = 1 microg/ml) against all strains tested with a minimal inhibitory concentration (MIC) determination ranging from 0.12 to 4 microg/ml. No resistant strains of A. pleuropneumoniae, P. multocida and M. haemolytica to FFC have apparently developed since the first introduction of this antibiotics for veterinary use in Korea. The results suggest that FFC is therapeutically valuable in the treatment of primary or complicating bacterial pathogens causing of the bovine and swine respiratory tract.     

我国动物源细菌耐药状况、存在问题和对策

动物源细菌耐药性问题不仅关系到动物的用药安全,也与公共卫生安全密切相关,日益引起各国的关注。中国兽医药品监察所从事动物源细菌耐药性检测工作10余年,对我国动物源细菌的耐药性状况进行了系统的调查。本文从动物源细菌耐药性监测工作的重要性、我国动物源细菌的耐药性状况、存在的问题以及应对策略四个方面进行了综述,以期对我国的动物源细菌耐药性监测工作起一定的参考作用。     

Antimicrobial susceptibility of Mycoplasma hyorhinis

A broth microdilution technique was used to determine the antimicrobial susceptibility of 15 field isolates of Mycoplasma hyorhinis to 10 antimicrobial agents, representative of different classes, and contrasting newer agents to existing ones. For the macrolides, the MIC(90) for tylosin and tilmicosin was 1 and 4 microg/ml, respectively, but was > or = 16 microg/ml for erythromycin. Tetracycline, lincomycin and enrofloxacin each had an MIC(90) of 2 microg/ml. The mycoplasma had similar levels of susceptibility to the aminoglycoside and aminocyclictol classes exhibiting an MIC(90) of 4 microg/ml for gentamicin and 2 microg/ml for spectinomycin. The isolates exhibited high MICs to trimethoprim/sulfamethoxazole with an MIC(90) > or = 16/304 microg/ml. In summary, M. hyorhinis isolates from the US had low MICs against a variety of antimicrobials tested, with the exception of erythromycin and trimethoprim/sulfamethoxazole.     

Susceptibility of different bacterial species isolated from food animals to copper sulphate, zinc chloride and antimicrobial substances used for disinfection

A total of 569 different bacterial isolates (156 Salmonella, 202 E. coli, 43 S. aureus, 38 S. hyicus, 52 E. faecalis, 78 E. faecium) were tested for susceptibility to copper sulphate, benzalkonium chloride, hydrogen peroxide and chlorhexidine using MIC determinations. A total of 442 isolates were also tested for susceptibility to formaldehyde and 177 isolates for susceptibility to zinc chloride. Enterococcal isolates formed a bimodal distribution of MICs to copper sulphate, whereas the other bacterial species formed one large population. Otherwise the isolates formed one large population of susceptibilities to the different antimicrobial agents. Large variations were observed in the susceptibility of the different bacterial species to the different compounds. Staphylococci were in general very susceptible to all antimicrobial compounds tested. The Salmonella isolates were in general less susceptible to copper sulphate, benzalkonium chloride and chlorhexidine followed by E. coli and the Gram-positive species. The opposite was the case for zinc chloride. All isolates were very susceptible to H(2)O(2) with MICs ranging from 0.002 to 0.016%, and to formaldehyde with MICs at 0.003 and 0.006%. This study showed that Danish bacterial isolates from livestock so far have not or have only to a limited degree developed resistance to antimicrobial compounds commonly used for disinfection. Acquired copper resistance was only found in enterococci. There were large differences in the intrinsic susceptibility of the different bacterial species to these compounds, and Salmonella especially seems intrinsically less susceptible than the other bacterial species, which might have human health implications.