Prevalence of metallo-beta-lactamase among Pseudomonas aeruginosa and Acinetobacter baumannii in a Korean university hospital and comparison of screening methods for detecting metallo-beta-lactamase

To identify the metallo-beta-lactamases (MBLs) prevalent in Korea, a total of 130 clinical isolates of Pseudomonas aeruginosa and Acinetobacter baumannii (99 P. aeruginosa and 31 A. baumannii) with a reduced susceptibility to imipenem (IPM) and/or ceftazidime (CAZ) was subjected to PCR analyses with primers specific to bla(IMP-1), bla(VIM-1), and bla(VIM-2). In addition, inhibitor-potentiated disk diffusion methods (IPD) using two kinds of substrate-inhibitor combinations (ceftazidime-2-mercaptopropionic acid (2MPA) and imipenem-EDTA) were investigated. Thirty-three isolates (29 P. aeruginosa and 4 A. baumannii) carried bla(VIM-2) and two P. aeruginosa isolates harbored bla(IMP-1). The enterobacterial repetitive intergenic consensus PCR (ERIC-PCR) pattern revealed that many of the VIM-2-producing P. aeruginosa isolates were clonally related, whereas the A. baumannii isolates were diverse. The inhibitor-potentiated disk diffusion test using imipenem-EDTA was highly sensitive and specific for detecting the VIM-2 producer. These results suggest that VIM-2 is an important MBL in P. aeruginosa and A. baumannii in the Korean hospital of this study and that the IMP-1-producing P. aeruginosa has also emerged. Screening for MBLs and strict infection control for these isolates will contribute to prevent further spread of resistance.     

Infectivity and resistance to antibiotics of bacterial strains isolated from patients hospitalised in intensive care units

The aim of this study was to evaluate the frequency of isolation and antimicrobial resistance testing of bacterial strains isolated from clinical specimens from patients hospitalized in three Intensive Care Units in Wroc?aw. The susceptibility of bacteria (107 strains) to selected antibiotics was determined. The results clearly show that non-fermentative rods were identified as the main agents causing pneumonia (58% of isolates). The second commonest pathogens were Gram-positive cocci (29%). The P. aeruginosa and E. cloacae strains were resistant to ampicillin, amoxicillin/clavulanate, cefuroxime and cefotaxime. All isolates of A. baumanii were susceptible only to imipenem. The rods of K. pneumoniae and E. coli were resistant to ampicillin, about 55% strains of both bacteria were sensitive to other antibiotics, except piperacillin/tazobactam, imipenem and ciprofloxacin. About 90% of methicillin resistant S. epidermidis strains were resistant to all antibiotics, except vancomycin (100% isolates were sensitive). ESBL were detected among E. cloace, K. pneumoniae and E. coli. We found P. aeruginosa rods producing MBL.     

铜绿假单胞菌对碳青霉烯类药物的耐药机制研究

目的 探讨铜绿假单胞菌对碳青霉烯类药物的耐药机制.方法 收集2008年11月至2009年4月我院临床分离的铜绿假单胞菌31株,根据药敏结果分为碳青霉烯类耐药组(21株)和碳青霉烯类敏感组(10株).另设1株标准株ATCC 27853,用亚胺培南-EDTA(乙二胺四乙酸)抑制试验检测菌株是否产生金屑酶,采用PCR法检测各菌株的外膜孔道蛋白oprD2基因,探讨铜绿假单胞菌对碳青霉烯类抗生素耐药机制.结果 21株耐药株有7株产生金属酶;21株耐药株经oprD2基因扩增,15株阴性,6株阳性,10株敏感株全部阳性.统计学检验结果表明,碳青霉烯类耐药组与敏感组oprD2基因阳性率的差异有极显著性(P＜0.01).结论 oprD2基因缺失和金属酶是本院铜绿假单胞菌对碳青霉烯类抗生素耐药的重要机制.     

Use of artificial sputum medium to test antibiotic efficacy against Pseudomonas aeruginosa in conditions more relevant to the cystic fibrosis lung

There is growing concern about the relevance of in vitro antimicrobial susceptibility tests when applied to isolates of P. aeruginosa from cystic fibrosis (CF) patients. Existing methods rely on single or a few isolates grown aerobically and planktonically. Predetermined cut-offs are used to define whether the bacteria are sensitive or resistant to any given antibiotic. However, during chronic lung infections in CF, P. aeruginosa populations exist in biofilms and there is evidence that the environment is largely microaerophilic. The stark difference in conditions between bacteria in the lung and those during diagnostic testing has called into question the reliability and even relevance of these tests. Artificial sputum medium (ASM) is a culture medium containing the components of CF patient sputum, including amino acids, mucin and free DNA. P. aeruginosa growth in ASM mimics growth during CF infections, with the formation of self-aggregating biofilm structures and population divergence. The aim of this study was to develop a microtitre-plate assay to study antimicrobial susceptibility of P. aeruginosa based on growth in ASM, which is applicable to both microaerophilic and aerobic conditions. An ASM assay was developed in a microtitre plate format. P. aeruginosa biofilms were allowed to develop for 3 days prior to incubation with antimicrobial agents at different concentrations for 24 hours. After biofilm disruption, cell viability was measured by staining with resazurin. This assay was used to ascertain the sessile cell minimum inhibitory concentration (SMIC) of tobramycin for 15 different P. aeruginosa isolates under aerobic and microaerophilic conditions and SMIC values were compared to those obtained with standard broth growth. Whilst there was some evidence for increased MIC values for isolates grown in ASM when compared to their planktonic counterparts, the biggest differences were found with bacteria tested in microaerophilic conditions, which showed a much increased resistance up to a > 128 fold, towards tobramycin in the ASM system when compared to assays carried out in aerobic conditions. The lack of association between current susceptibility testing methods and clinical outcome has questioned the validity of current methods. Several in vitro models have been used previously to study P. aeruginosa biofilms. However, these methods rely on surface attached biofilms, whereas the ASM biofilms resemble those observed in the CF lung. In addition, reduced oxygen concentration in the mucus has been shown to alter the behavior of P. aeruginosa and affect antibiotic susceptibility. Therefore using ASM under microaerophilic conditions may provide a more realistic environment in which to study antimicrobial susceptibility.     

2014—2016年某院铜绿假单胞菌耐药性监测

摘要：目的 分析铜绿假单胞菌的分布和耐药性变化,为临床防治铜绿假单胞菌感染提供依据。方法 收集成都大学附属医院2014—2016年所分离的铜绿假单胞菌,采用VITEK 2-Compact全自动细菌鉴定系统进行鉴定和药敏试验,采用WHONET 5.6软件对数据进行分析。结果 3年间共分离出1 945株铜绿假单胞菌,多重耐药铜绿假单胞菌分离率为34.2%（666/1945）。标本来源以呼吸道为主,占82.2%（1598/1945）。科室分布以呼吸内科最多,ICU其次。铜绿假单胞菌对头孢曲松和头孢噻肟的耐药率最高,均＞57.0%;对阿米卡星耐药率最低,为2.0%。3年来铜绿假单胞菌对17种抗生素的耐药率呈整体上升的趋势。结论 铜绿假单胞菌对头孢曲松、头孢噻肟、亚胺培南耐药率较高,对阿米卡星耐药率较低。铜绿假单胞菌的耐药率呈整体上升的趋势,应重视细菌耐药性的监测,以延缓耐药性产生、促进临床合理用药。     

铜绿假单胞菌的院内感染分布及耐药性的调查

目的了解铜绿假单胞菌（PA）在医院内感染的分布特点及其耐药性,指导临床合理用药,降低医院感染率。方法回顾分析2005年1月至2008年11月我院临床分离的铜绿假单胞菌152株,按WHO推荐的NC-CLS标准方法（K—B法）进行药敏试验,分析菌株的临床分布特征及体外药敏结果。结果铜绿假单胞菌在临床上主要以呼吸道感染为主。在17种临床常用抗生素中,多表现为多重耐药。美洛培南、亚胺培南及左旋氧氟沙星抗PA效果最好,耐药率分别只有14．47％、14．47和5．26％。结论多重耐药铜绿假单胞菌在临床广泛存在,应当根据临床体外药敏结果合理选择抗菌药物进行治疗,以取得良好临床效果并减少耐药菌产生;同时规范院内感染控制措施,减少耐药菌株的播散。     

Iodine sensitivity of bacteria isolated from iodinated water systems

Fourteen bacterial isolates, predominantly Pseudomonas sp., from two water systems disinfected by iodinated anion-exchange resins were studied and compared with an isolate of Pseudomonas aeruginosa from a povidone-iodine solution and four other isolates. Pseudomonas cepacia and P. aeruginosa grown in brain heart infusion were 3 to 5 logs less sensitive to 1 mg/L I2 (pH 7.2, 1 min) when compared with cultures grown in phosphate buffer. Another P. cepacia isolate was the least sensitive culture when grown in brain heart infusion (1 log decrease) but was more sensitive after cultivation in phosphate buffer (5 logs). Isolates from an iodinated potable water system, including P. cepacia, Staphyloccus warneri, and a Bacillus sp., were all less sensitive to iodine than a "resistant" P. aeruginosa and three other isolates when grown in brain heart infusion. A clinical isolate of P. aeruginosa exhibited intermediate sensitivity. The sensitivity of bacteria to iodine is thus highly variable, depending on the organism as well as the growth conditions.     

Antibiotic susceptibility and occurrence of ESBL, IBL and MBL in Pseudomonas aeruginosa strains

The aim of this study was to evaluate the drug susceptibility of P. aeruginosa strains and to detect strains producing inducible beta-lactamases (IBL), extended-spectrum beta-lactamases (ESBL), and metallo-beta-lactamases (MBL). During 6 month (October 2005 - March 2006), 66 strains of P. aeruginosa strains were cultured from clinical specimens obtained from patients of two of hospitals in Siedlce and from patients of outpatient clinics. All the strains were identified in the automatic ATB (bio Mérieux). The susceptibility of bacteria to antibiotics was tested by standard disc diffusion method. The majority of strains were susceptible to meropenem (89.4%), piperacillin combined with tazobactam (84.8%), ciprofloxacin (84.8%) and piperacillin (83.3%). Many of our strains were resistant to carbenicillin (69.7%), mezlocillin (45.5%), gentamicin (42.4%) and netylmicin (30.3%). 6 strains (9.1%) were multidrug-resistant (MDR). Inducible beta-lactamases were detected with the use double disc method according to Sanders and Sanders. ESBL-producing strains were detected with double disc test (DDST) according to Jarlier et al. These strains were identified as ESBL-positive on the basis of the DDST were also determined using a double disc (DD) test according to Appleton. Production of metallo-beta-lactamases (MBL) was examined with the use of Etest MBL (AB Biodisk, Sweden) and the double disc test according to Arakava et al. Sixty-five IBL-producing strains (98.5% of all strains) and three strains (4.5%) with MBL activity were detected. Strains producing extended beta-lactamases (ESBL) were not found.     

Antimicrobial susceptibility and molecular mechanisms of resistance to beta-lactams of gram-negative microorganisms--causative agents of nosocomial infections

Profiles and mechanisms of resistance to beta-lactam antibiotics of isolates of Gram-negative microorganisms, which are causative agents of infections in Intensive Care Unit of hospital surgery department, were studied. Two hundred and ten clinical isolates were studied: Pseudomonas aeruginosa--86 strains (40.9%), Acinetobacter baummanii--45 strains (21.4%), Klebsiella pneumoniae--52 strains (24.8%), Escherichia coli--23 strains (11%), Enterobacter spp.--4 strains (1.9%). Profiles of antibiotic resistance were studied by the method of serial microdilutions; detection of most widespread and clinically significant genes of beta-lactamases of Gram-negative bacteria was performed by polymerase chain reaction. Carbapenems and cefoperazone/sulbactam were the most active antibiotics. Local features of distribution of beta-lactamase coding genes (TEM, SHV, CTX) in K. pneumoniae and E. coli isolates were revealed. Eleven strains of P. aeruginosa resistant to carbapenems and possessing genetic determinants of VIM-group, which codes metallo-beta-lactamases, were isolated. Obtained data allows to assess the parameters of resistance to beta-lactam antibiotics and to reveal the main mechanisms of such resistance in etiologic agents of nosocomial infections, that, in its turn, allows to choose preparations for etiotropic therapy.     

Frequency and susceptibility patterns of non-fermenting gram-negative rods isolated from patients hospitalised in intensive care units of a tertiary care hospital

The aim of the study was to assess frequency and susceptibility to antimicrobial agents of non-fermenting gram-negative rods isolated from clinical specimens obtained from patients requiring intensive care, with emphasis on profile of the unit. Identification of cultured isolates was done using automated VITEK and API systems (bioMerieux, France). Susceptibility to antimicrobial agents was tested by a disk-diffusion method according to the NCCLS recommendations. In total the analysis comprised 425 strains of non-fermenting gram-negative rods, constituting 58.9% of all isolates of gram-negative bacteria. In blood cultures predominated strains of A. baumannii (46.8%) and P. aeruginosa (40.4%), while in cultures of other clinical specimens these bacteria comprised 42.9% and 43.9% of isolates. Major differences were observed in frequency of these species on both ICU units. Strains of non-fermenting rods isolated from blood cultures comprised a lower percentage of strains susceptible to antimicrobials (particularly cefepime and carbapenems) than isolates cultured from other specimens. Strains of A. baumannii resistant to imipenem and meropenem were detected with a frequency of 12.5% and 26.7%, respectively. Resistance of P. aeruginosa strains to carbapenems was 62.2% and 44.3%, respectively. There was a relatively high percentage of strains susceptible to cefepime (82.0%), ceftazidime (78.9%), amikacin (77.8%) and piperacillin/tazobactam (69.7%). Conclusions: 1. There was a predominance (58.9%) of strains of gram-negative non-fermenting rods. 2. Isolates from blood cultures were characterised by a much higher percentage of resistant strains in comparison to other specimens. 3. Strains of A. baumannii resistant to carbapenems were recorded. 4. There were differences in frequency and antimicrobial susceptibility among the strains of P. aeruginosa and A. baumannii depending on the type of clinical specimen and ICU profile.     

Identification and genetic characterization of metallo-beta-lactamase-producing strains of Pseudomonas aeruginosa in Tehran, Iran

Metallo-beta-lactamases (MBLs) are being reported with increasing frequency worldwide. The aim of this study was to investigate the prevalence of blalMP-1, blaVIM-1,2 and blaSPM-1 genes encoding metallo-beta-lactamases (MBLs) among a collection of Pseudomonas aeruginosa strains isolated from patients at different hospitals in Tehran and to trace the disseminated clones at these hospitals by pulsed field gel electrophoresis (PFGE). Susceptibility of 610 P aeruginosa to 14 different antibiotics was determined using disc diffusion method. Isolates showing resistance to imipenem and ceftazidime were subjected to micro broth dilution assay to determine their MIC values. The blaIMP-1, blaVIM-1, blaVIM-2, and blaSPM-1, genes were amplified by PCR. Isolates containing blaVIM-1 were analyzed by PFGE. Sixty-eight isolates were resistant to imipenem (MIC > or = 4 microg/ml) of which 16 isolates carried blaVIM-1 gene using PCR assay. No other MBL genes were detected in this study. Three different unrelated patterns were found for isolates containing blaVIM-1 gene by PFGE of which pattern A was predominant. All isolates were susceptible to colistin and polymixin B. blaVIM-1 was the main gene encoding MBL among the isolates of P aeruginosa in our study. Clonal spread of isolates containing blaVIM-1 had occurred at Tehran hospitals. However, heterogeneous clones also were involved in the outbreaks.     

Antibiotic sensitivity pattern; experience at University Hospital, Riyadh, Saudi Arabia

Results of sensitivity testing were discussed based on examination of 5192 isolates of the various bacteria isolated from clinical specimens from King Khalid University Hospital in Riyadh, Saudi Arabia. Streptococcus pyogenes and Streptococcus pneumoniae were sensitive to penicillin and erythromycin. The sensitivity pattern of Staphylococcus aureus was also predictable as they were fairly sensitive to both methicillin (98%) and erythromycin (96%). Neisseria gonorrhoeae (27%) showed a high level of resistance to penicillin. The resistance of Haemophilus influenzae to ampicillin and chloramphenicol was low. Brucella species was sensitive to tetracycline and rifampicin; resistance to streptomycin and cotrimoxazole was minimal being 1% and 6% respectively. The resistance of E. coli, Klebsiella species and Proteus species to second and third generation cephalosporins and amikacin was fairly low ranging from 1.3% to 3%. The gentamicin resistance for these organisms was also within the acceptable range (3%-10%). Gentamicin and amikacin resistance for Pseudomonas aeruginosa was low (2-8%). Salmonella typhi was sensitive to ampicillin, cotrimoxazole, and chloramphenicol. Salmonella enteritidis, Shigella species, and enteropathogenic E. coli were highly resistant to various antibiotics. Campylobacter jejuni was sensitive to gentamicin but 6% of isolates were resistant to erythromycin. Ninety six percent of Gram-negative rods except P. aeruginosa isolated from urine of patients having urinary tract infections were sensitive to amoxycillin-clavulanic acid. In addition, P. aeruginosa showed fairly low resistance to norfloxacin which is given orally to treat cystitis caused by this organism.     

Effect of age and housing location on antibiotic resistance of fecal coliforms from pigs in a non-antibiotic-exposed herd.

The relationship of age and housing location to single antibiotic resistance, multiple antibiotic resistance, and resistance patterns of fecal coliforms obtained during a 20-month period from pigs in a herd that was not exposed to antibiotics for 126 months was determined. Bacteria resistant to single and multiple antibiotics were isolated more frequently (P less than 0.01) from pigs under 7 months of age. A greater proportion of isolates from pigs over 6 months of age was sensitive to the 13 antimicrobial agents tested (P less than 0.01), while a smaller proportion showed resistance to single (P less than 0.05) and multiple (P less than 0.01) antibiotics. More than 80% of the resistant isolates were resistant to tetracycline, streptomycin, or sulfisoxazole. Resistance was greater (P less than 0.01) for pigs in the finishing unit than for those on pasture. Resistance to ampicillin, carbenicillin, and tetracycline was greater (P less than 0.05) for pigs in the finishing unit than for those in the farrowing house. More isolates from pigs on pasture were sensitive to all antimicrobial agents tested (P less than 0.01). A greater proportion of isolates from pigs in the finishing unit showed resistance to a single antibiotic (P less than 0.01). The data from this study suggest that exposure to antibiotics is not the only factor that influences the prevalence of bacteria that are resistant to single and multiple antibiotics in the feces of domestic animals and that considerable research is needed to define the factors influencing antibiotic resistance in fecal bacteria.     

Effect of age and housing location on antibiotic resistance of fecal coliforms from pigs in a non-antibiotic-exposed herd

The relationship of age and housing location to single antibiotic resistance, multiple antibiotic resistance, and resistance patterns of fecal coliforms obtained during a 20-month period from pigs in a herd that was not exposed to antibiotics for 126 months was determined. Bacteria resistant to single and multiple antibiotics were isolated more frequently (P less than 0.01) from pigs under 7 months of age. A greater proportion of isolates from pigs over 6 months of age was sensitive to the 13 antimicrobial agents tested (P less than 0.01), while a smaller proportion showed resistance to single (P less than 0.05) and multiple (P less than 0.01) antibiotics. More than 80% of the resistant isolates were resistant to tetracycline, streptomycin, or sulfisoxazole. Resistance was greater (P less than 0.01) for pigs in the finishing unit than for those on pasture. Resistance to ampicillin, carbenicillin, and tetracycline was greater (P less than 0.05) for pigs in the finishing unit than for those in the farrowing house. More isolates from pigs on pasture were sensitive to all antimicrobial agents tested (P less than 0.01). A greater proportion of isolates from pigs in the finishing unit showed resistance to a single antibiotic (P less than 0.01). The data from this study suggest that exposure to antibiotics is not the only factor that influences the prevalence of bacteria that are resistant to single and multiple antibiotics in the feces of domestic animals and that considerable research is needed to define the factors influencing antibiotic resistance in fecal bacteria.     

圈养林麝铜绿假单胞菌的分离鉴定及耐药性和病原特性分析

【背景】铜绿假单胞菌感染所致的化脓性疾病是困扰林麝驯养的重要因素,是一类较难防治的细菌性疾病,目前尚无疫苗可用来预防该病。【目的】研究林麝源铜绿假单胞菌的感染现状和分子流行病学规律。【方法】对2014年10月至2015年10月四川宝兴和陕西镇坪两个林麝养殖中心发病林麝中铜绿假单胞菌进行分离鉴定,并对其耐药情况进行分析,利用脉冲场凝胶电泳(PFGE)分型技术研究分离菌的PFGE指纹图谱,探究其流行病学趋势,并对部分菌株的致病性进行分析。【结果】分离得到60株铜绿假单胞菌,其中34株来自镇坪,26株来自宝兴。耐药结果表明:60株林麝源铜绿假单胞菌对17种抗菌药物呈现不同程度耐药性,不同地区和不同样本源间分离的铜绿假单胞菌对17种抗菌药物的耐药性总体趋于一致,多重耐药情况严重,以5耐、6耐为主。分型结果显示:60株铜绿假单胞菌PFGE图谱的相似性为49.1%-100%。经聚类分析得到A-O共15种基因簇,其中优势基因簇为C、E、G、J。致病性结果表明,流行菌株的致病力强弱依次为:动物源菌株环境源菌株,且主要流行菌株(基因簇E、F、J)的致病力大于其余菌群。【结论】铜绿假单胞菌在两地区具有水平传播的途径,本研究可为跨地区林麝化脓性炎症的防控提供理论依据。     

Susceptibility to chemotherapeutics and the occurrence of metallo-beta-lactamases in hospital Pseudomonas aeruginosa strains

The aim of the study was evaluation of susceptibility of Pseudomonas aeruginosa strains isolated from patients hospitalized in different wards of Rydygier's Hospital in Krakow in 2005. Bacteria were identified on the basis of typical morphology confirmed by Gram-staining microscopy and by biochemical tests--ID 32 GN strips using ATB system (bioMerieux, France). The susceptibility of all isolates to a panel of antimicrobial agents was performed using disk diffusion method. The highest in vitro activity against clinical strains demonstrated ceftazidime (88.6% of susceptible strains) while the lowest in vitro activity against clinical strains demonstrated imipenem (50.4% of susceptible strains). It was also observed that 40.2%(53) of strains were resistant to meropenem and imipenem. Carbapenem resistant P aeruginosa strains were tested for MBL production. We performed disk synergy test for MBL detection with EDTA, 2-MPA and ceftazidime, imipenem. The presence of a distorted inhibition zone was interpreted as a positive result for MBL production. Positive results of disk synergy tests were confirmed by Etest MBL strips. Metalo-beta-lactamases were detected in 13 isolates resistant to carbapenems.     

铜绿假单胞菌医院感染分布及耐药性分析

目的探讨铜绿假单胞菌临床分离株的感染分布情况及其抗生素的耐药情况,指导临床合理用药。方法对我院2006年1月至2008年6月住院患者分离的铜绿假单胞菌进行抗生素敏感性测定,采用API系统及VITEK2系统进行细菌鉴定,用K-B法进行药敏试验及结果观察。结果216例院内感染铜绿假单胞菌病例中,从痰标本中分离的菌株最多,阳性率为45．4％;铜绿假单胞菌对头孢噻肟、复方新诺明、头孢哌酮的耐药率较高,分别为88％、86．1％和85．2％,对美洛培南的敏感性最高达到94％。结论铜绿假单胞菌是医院病原菌感染的主要致病菌之一,加强细菌和药敏监测,选择敏感性强的药物,避免广谱抗菌药物的长期应用,是延缓耐药菌株增加的有效办法。     

Multiniche screening reveals the clinically relevant metallo-beta-lactamase VIM-2 in Pseudomonas aeruginosa far from the hospital setting: an ongoing dispersion process?

A screening study of the presence of metallo-beta-lactamases (IMP and VIM types and SPM-1) in isolates from different nonhospital sources was conducted, and it revealed the presence of bla(VIM-2), associated with the In58 class 1 integron, in two unrelated Pseudomonas aeruginosa strains from aquatic habitats. The results suggest that the hospital setting was the possible origin of these bla(VIM-2)-carrying strains.     

Susceptibility of Pseudomonas aeruginosa to Gentamicin Sulfate In Vitro: Lack of Correlation Between Disc Diffusion and Broth Dilution Sensitivity Data

Seventy-eight of 420 clinical isolates of Pseudomonas aeruginosa yielded zones of inhibition of less than 12 mm in diameter around 10-mug discs of gentamicin sulfate when tested by the standardized Bauer-Kirby disc diffusion method. Of 153 strains chosen from these isolates, one strain (0.65%) required 25 mug of gentamicin per ml for inhibition; the remainder (99.35%) were inhibited by 6 mug/ml or less of the antibiotic. It is recommended that those isolates of P. aeruginosa that yield zones of inhibition less than 12 mm in diameter be disc susceptibility-tested once more; those isolates that give zones of inhibition of less than 12 mm upon repeated examination should then be subjected to the broth dilution test before they are designated as sensitive or resistant to gentamicin.     

Antibiotic resistance of gram-negative enteric bacteria from pigs in three herds with different histories of antibiotic exposure

The antibiotic resistance patterns of gram-negative fecal bacteria from pigs in three herds with different histories of antibiotic exposure were examined. In general, smaller proportions of antibiotic-resistant or multiply resistant fecal isolates (P less than 0.05) were obtained from pigs in a herd not exposed to antimicrobial agents for 154 months than from pigs in a herd continuously exposed to antimicrobial agents at subtherapeutic doses or from pigs in a herd exposed only to therapeutic doses of antimicrobial agents. The proportions of antibiotic-resistant and multiply resistant strains were greater among isolates from pigs in the therapeutic herd than in the non-antibiotic-exposed herd (P less than 0.05). The proportion of antibiotic-resistant isolates in the non-lactose-fermenting population was greater than that in the lactose-fermenting population, regardless of herd. The results suggest that any form of antimicrobial exposure will increase the prevalence of antimicrobial resistance and multiple resistance of fecal bacteria.