Minimal inhibitory concentrations of antimicrobial agents against Actinobacillus pleuropneumoniae.

Forty-five isolates of Actinobacillus pleuropneumoniae were tested for susceptibility to 12 antimicrobial agents using a microdilution method for the minimal inhibitory concentration determinations. These results confirmed the high prevalence of A. pleuropneumoniae strains resistant to antibiotics as reported earlier using the disc diffusion method (Kirby-Bauer method). While 36% of the isolates were resistant to the penicillins, 47% were resistant to chloramphenicol and 68% were resistant to tetracycline. Minimal inhibitory concentrations for the resistant isolates were approximately 32 times higher than those for the susceptible isolates to the above antibacterial agents. The isolates were in general weakly susceptible or resistant to spectinomycin, lincomycin, tiamulin and spiramycin whereas most of them were susceptible to gentamicin, trimethoprim and erythromycin. The susceptibility pattern was similar throughout the 1980 to 1984 period. The 14 serotype 5 isolates were more resistant to tetracycline but less resistant to chloramphenicol and the penicillins than the 28 serotype 1 isolates.     

Minimum inhibitory concentrations of 20 antimicrobial agents against Staphylococcus aureus isolated from bovine intramammary infections in Japan

Minimum inhibitory concentrations (MICs) of 20 antimicrobial agents were determined against 51 isolates of Staphylococcus aureus from bovine intramammary infections. Fourteen (27.4%) isolates were resistant to benzylpenicillin, but none of the isolates was resistant to cloxacillin, nafcillin, or cephems. Among aminoglycosides, gentamicin was the most active, with an MIC50 of 0.2 microg/ml, followed by kanamycin, with an MIC50 of 0.78 microg/ml. Five isolates (9.8%) were resistant to dihydrostreptomycin, three isolates (5.9%) to kanamycin and two isolates (3.9%) to gentamicin. Resistance to erythromycin was observed in two isolates (3.9%). Tylosin was less active than erythromycin, with MIC50s of 1.56 microg/ml versus 0.39 microg/ml, but none of the isolates was resistant to this antibiotic. Oxytetracycline MICs were situated in the range of 0.39-1.56 microg/ml for 48 susceptible isolates. Although 19 (37.3%) isolates were resistant to one or more antimicrobial agents, a single resistance pattern was most frequent: benzylpenicillin (12 isolates), dihydrostreptomycin (two isolates) and kanamycin (one isolate). There were no isolates resistant to antimicrobial agents such as methicillin, lincomycin, clindamycin, chloramphenicol, florfenicol and virginiamycin, which have not been approved for use in cattle husbandry in Japan.     

Antimicrobial resistance of bacteria isolated from dairy cow milk samples submitted for bacterial culture: 8,905 samples (1994-2001)

OBJECTIVE: To determine whether antimicrobial resistance patterns of major mastitis pathogens isolated from milk samples from dairy cows have changed over time. DESIGN: Retrospective study. SAMPLE POPULATION: 8905 bacterial isolates obtained from milk samples submitted to the Wisconsin Veterinary Diagnostic Laboratory between January 1994 and June 2001. PROCEDURE: Antimicrobial susceptibility was determined by means of the Kirby-Bauer disk diffusion method. Logistic regression was used to determine whether percentages of isolates resistant to various antimicrobials changed over time. RESULTS: For the gram-positive mastitis pathogens, percentages of isolates resistant to various flactam antimicrobials did not increase over the course of the study. Percentage of Staphylococcus aureus isolates resistant to penicillin decreased from 49 to 30%; percentage of Streptococcus isolates resistant to penicillin decreased from 6 to 1%. Percentage of isolates resistant to erythromycin increased for S aureus, Escherichia coli, Enterobacter spp, Enterococcus spp, and Pasteurella spp. Percentage of isolates resistant to lincomycin increased for S aureus and Staphylococcus spp. Percentage of coagulase-negative Staphylococcus isolates resistant to pirlimycin increased from 6 to 19%. For several pathogens, percentages of isolates resistant to sulfisoxazole and to trimethoprim-sulfamethoxazole decreased. No pathogens had a significant increase in the percentage of isolates resistant to novobiocin-penicillin. CONCLUSIONS AND CLINICAL RELEVANCE: Results did not indicate a trend toward increased antimicrobial resistance among mastitis pathogens isolated from milk samples from dairy cows between 1994 and 2001. Reduced resistance to flactam antimicrobials was identified for several gram-positive mastitis pathogens.     

Antimicrobial resistance of Escherichia coli O26 and O111 isolates from cattle and their characteristics

The present study was to investigate antimicrobial resistance profiles of Escherichia coli O26 and O111 from cattle and to characterize the virulence genes of the resistant isolates. This paper reports the high prevalence of antimicrobial resistant E. coli O26 and O111 from cattle. Among 37 E. coli O26 and 25 E. coli O111 isolates from the fecal specimens obtained from cattle, 26 (70%) and 15 (60%) were resistant to at least one antibiotic, respectively. Forty (98%) of the 41 resistant isolates were resistant to two or more antibiotics. Among the 22 antibiotics tested in this study, ampicillin was the most common antibiotic that the isolates were resistant to, followed by tetracycline and streptomycin. None of the isolates were resistant to fluoroquinolones, such as ciprofloxacin, ofloxacin and norfloxacin, and to ceftriaxone, amikacin and imipenem. Eighteen different resistant types among the 41 isolates were observed by the cluster analysis. The most frequent antibiotic-resistance type was ampicillin-tetracycline-streptomycin-cephalothin-sulfisoxazole-ticarcillin-kanamycin-minocycline-piperacillin-chloramphenicol, which accounted for 9 (22%) of the resistant isolates. The observation of frequent and multiple resistances to antibiotics highlights the need for their careful use if their benefits are to be preserved. PCR analysis of the EHEC virulence markers showed that 25 of the resistant E. coli O26 and O111 isolates tested positive for stx2 or both stx1 and stx2. This suggests that the majority of these isolates can cause serious diseases in humans and may complicate the future therapeutic options under development.     

Trends in the resistance to antimicrobial agents of Streptococcus suis isolates from Denmark and Sweden

This study was conducted to determine the MIC values of historical and contemporary Streptoccocus suis (serotypes 2 and 7) from Denmark and S. suis (serotype 2) from Sweden. A total of 52 isolates originating from 1967 through 1981 and 156 isolates from 1992 through 1997 in Denmark and 13 isolates from Sweden were examined for their MICs against 20 different antimicrobial agents. Most antimicrobials were active against most isolates. A frequent occurrence of resistance to sulphamethoxazole was observed, with most resistance among historic isolates of serotype 7 and least resistance among isolates from Sweden. A large number of the isolates was resistant to macrolides. However, all historic serotype 2 isolates from Denmark were susceptible, whereas 20.4% of the contemporary isolates were resistant. Among serotype 7 isolates 23.3% of the historic isolates were resistant to macrolides, whereas resistance was found in 44.8% of the contemporary isolates. All isolates from Sweden were susceptible to macrolides. Time-associated frequency of resistance to tetracycline was also found. Only a single historic isolate of serotype 2 was resistant to tetracycline, whereas 43.9% of the contemporary serotype 2 isolates and 15.5% of the contemporary serotype 7 isolates were resistant. Only one (7.7%) of the isolates from Sweden was resistant. The differences in resistance between historic and contemporary isolates from Denmark were statistically significant. This study demonstrated a significant serotype-associated difference in the susceptibility to macrolides and tetracycline and demonstrated that an increase in resistance among S. suis isolates has taken place during the last 15 years to the two most commonly used antimicrobial agents (tylosin and tetracycline) in pig production in Denmark.     

Phenotypic and Genotypic Anti‐Microbial Resistance Profiles of Campylobacters from Untreated Feedlot Cattle and Their Environment

Anti‐microbial resistance is an emerging public health issue. Farmed animals may act as reservoirs and potential sources of anti‐microbial resistant Campylobacters. The aim of this study was to investigate the anti‐microbial resistance profile of cattle and environmental Campylobacter isolates from normal untreated feedlot cattle, the role of the gyrA Thr‐86‐Ile mutation in ciprofloxacin‐resistant Campylobacter jejuni isolates and the involvement of the tripartite CmeABC efflux system for multi‐resistant C. jejuni isolates. The phenotypic anti‐microbial resistance testing was carried out on 500 Campylobacter isolates (445 cattle isolates and 55 environmental isolates). In general, there was a higher level of anti‐microbial resistance for the environmental isolates compared with the animal isolates, 45% of the animal isolates were resistant to one or more of the seven anti‐microbials compared with 84% of the environmental isolates. The combined cattle and environmental Campylobacters had 34 (6.8%) isolates resistant to three or more of the seven anti‐microbials tested on all isolates and 11 (2.2%) isolates were resistant to the seven anti‐microbials. There was a substantial level of ciprofloxacin‐resistant Campylobacters in both animal (8.5%) and environmental (21.8%) isolates. The gyrA Thr‐86‐Ile mutation was only present in five of 22 ciprofloxacin‐resistant C. jejuni isolates investigated. No multi‐drug‐resistant associated mutation was detected in the CmeB or the CmeR regions investigated. In conclusion, our study observed a substantial level of Campylobacter anti‐microbial resistance, highlighting the need for an active anti‐microbial surveillance program for food animals in Ireland and the importance of the chosen sampling point can have on the findings of such a program.     

奶牛乳房炎凝固酶阴性葡萄球菌耐药性分析

本研究对江苏部分奶牛场致隐性乳房炎病原进行流行病学调查,采样并进行细菌分离培养、革兰氏染色镜检、生化鉴定,共分离到35株凝固酶阴性葡萄球菌（CNS）。用常规药敏纸片法对CNS分离菌株的抗生素耐药情况进行分析,结果显示分离培养到的35株CNS对临床常用抗生素万古霉素、左氟沙星、克拉霉素、苯唑西林、诺氟沙星、氯霉素、环丙沙星、头孢噻肟、磷霉素和呋喃妥因等高度敏感,高敏率在60%～80%之间;而对青霉素G、复方新诺明和多黏菌素B有一定的耐药性,耐药率为70%～100%。分离菌株表现了较强的多重耐药性,耐20种药物的细菌有3株,耐15种药物的细菌有1株,耐14种药物的细菌有1株,62.9%的CNS分离菌株能耐5种以上抗生素,表明导致奶牛隐性乳房炎CNS菌株耐药情况不容小视。     

Antimicrobial resistance in generic fecal Escherichia coli from 29 beef farms in Ontario

The occurrence of antimicrobial resistance in generic Escherichia coli can serve as an indicator of the pool of resistance genes potentially available for transfer to pathogenic organisms. This study was conducted on 29 volunteer beef farms in Ontario to describe the prevalence and patterns of antimicrobial resistance in E. coli, and to describe changes in the prevalence of resistance during the feedlot stage of production. From the pooled fecal samples on 28 of the 29 farms, 31% of isolates from feedlots (n = 993) and 12% of isolates from cow-calf farms (n = 807) were resistant to one or more of 16 antimicrobials tested. No isolates were resistant to ceftriaxone, ciprofloxacin, gentamicin, or nalidixic acid, and < 1% of the pooled isolates were resistant to ceftiofur. Two percent of both feedlot and cow-calf isolates were resistant to > or = 5 antimicrobials. Cow-calf farms were at significantly lower risk than feedlots for having E. coli isolates that were resistant to streptomycin, sulfamethoxazole, and tetracycline. On average, the prevalence of sulfamethoxazole resistant E. coli isolates was significantly higher in calves than in cows. No resistance was observed to ceftriaxone or ciprofloxacin among isolates (n = 1265) obtained from individually sampled feedlot animals on 2 farms. Less than 1% of these isolates were resistant to gentamicin, nalidixic acid, and ceftiofur. From the individually sampled feedlot animals, resistance to streptomycin (on 1 farm), sulfamethoxazole, and tetracycline increased significantly from arrival to mid-point during the feeding period, and these levels persisted until market-readiness.     

禽源致病性沙门氏菌耐药表型与耐药基因的分析

为了解江苏苏中地区禽源沙门氏菌耐药现象及耐药基因的存在情况,采用K-B法对22株沙门氏菌分离株进行12种抗菌药物敏感性试验,并通过PCR方法检测了12种耐药基因的存在情况。结果显示,所有菌株对甲氧胺嘧啶、阿米卡星、庆大霉素、萘啶酮酸、诺氟沙星的耐药率均在72.73%～100%之间;存在着5种耐药表型,每个耐药表型均在4重或4重以上,耐药最多菌株可耐受10种药物;扩增出9种耐药基因,与GenBank中的参考序列同源性均在98%以上;耐药表型与耐药基因的存在基本一致,但无绝对相关性。本研究结果表明苏中地区沙门氏菌耐药现象非常严重,且耐药基因广泛存在。     

Microbial Safety of Chickens Raised Without Antibiotics

The purpose of the current study was to verify whether there was any validity to the claim that consumers could reduce their exposure to antibiotic-resistant bacteria by purchasing poultry products that were produced without antibiotics. Rinse samples were taken from whole carcasses from chickens grown in small flocks and marketed as antibiotic-free. Salmonella bacteria were isolated from 18.7% of all the carcasses sampled. Campylobacter bacteria were found on 96.0% of the carcasses tested. More than two-thirds (69.4%) of the Salmonella isolates were resistant to at least 1 antibiotic. The antibiotic to which the Salmonella isolates were most commonly resistant was trimethoprim-sulfa, with almost two-thirds (62.9%) of the isolates being resistant to it. Almost three-fourths (73.4%) of the Campylobacter isolates were resistant to at least 1 antibiotic. The most common antibiotic to which Campylobacter isolates were resistant was tetracycline, with almost three-fourths (72.7%) of all Campylobacter isolates being resistant to it.     

Antimicrobial susceptibility and presence of resistance genes in staphylococci from poultry

The species distribution, susceptibility to 19 antimicrobial agents and presence of selected genes encoding resistance to macrolides, streptogramins and tetracyclines were examined among 118 staphylococcal isolates from infections of poultry in Denmark. Isolates were identified using a combination of conventional biochemical testing and 16S rDNA sequencing. The most common species were Staphylococcus aureus (83), Staphylococcus hyicus (11), Staphylococcus xylosus (9) and Staphylococcus cohnii (6). The isolates were susceptible to most antimicrobials tested. A high frequency of S. aureus (30%) was resistant to ciprofloxacin. Only six (7%) S. aureus isolates and one Staphylococcus saprophyticus were penicillin resistant. Resistance to sulphamethoxazole was observed among 16 (19%) of S. aureus isolates and two coagulase negative staphylococci (CNS). Twenty (24%) of the S. aureus isolates were resistant to erythromycin and 19 of these isolates contained the ermA gene, whereas the remaining isolate contained the ermC gene. Eleven (48%) of the novobiocin resistant CNS were resistant to erythromycin and all these isolates contained the ermA gene. Two isolates identified as S. xylosus, were found to be resistant to streptogramins and both contained the vatB- and the vgaB-genes. Thirty-nine (47%) of the S. aureus isolates, three of nine S. hyicus and eight of the 23 novobiocin resistant CNS were tetracycline resistant and all contained the tet(K) gene. A single S. aureus isolate also contained the tet(M) gene. The present study showed a frequent occurrence of resistance to fluoroquinolones, tetracycline and macrolides among staphylococci isolated from broilers in Denmark, whereas the occurrence of resistance to other antimicrobial agents remains low. Similar genes, encoding resistance to erythromycin, tetracycline and streptogramins to those previously observed, were detected.     

我国牛源金黄色葡萄球菌耐药现状及药敏检测方法探讨

为调查我国牛源金黄色葡萄球菌的临床耐药现状,对2009年以来新疆、浙江、山东、内蒙古和上海五省区不同地方分离的120株金黄色葡萄球菌用纸片扩散法进行临床药敏试验。结果显示,新疆分离株对红霉素、克林霉素、青霉素、复方新诺明、多西环素、四环素耐药,22%的菌株对头孢西丁耐药,43%的对氯霉素耐药,64%的对环丙沙星耐药,58%的对庆大霉素耐药;其它4地区分离株耐药情况更为严重,除头孢西丁以外,对其它9种抗生素均耐药。对菌株的多重耐药情况分析发现,新疆分离株中对5种以上抗生素耐药的占86.1%,对10种抗生素完全耐药的占17.4%;而内地分离株的数据为100%和38.2%。此外,在试验中鉴定出33株耐甲氧西林金黄色葡萄球菌,新疆19株,占新疆菌株的22.1%,内地四地区14株,占内地分离株的41.2%。结合2005~2009年间发表的相关耐药数据的分析结果,研究显示,我国牛源金黄色葡萄球菌呈现多重耐药,且出现了耐甲氧西林金黄色葡萄球菌,并在不同地区分离株中占有一定的比例;研究还发现,新疆分离株与浙江、山东、内蒙古和上海四地分离株的耐药情况有较大差异。此外,在整理药敏试验资料时,发现在金黄色葡萄球菌药敏试验中,药物的选择没有遵循选药规则,试验操作不规范和试验结果的报告不规范等问题,对此提出一些探讨性建议,供试验人员参考。     

Antimicrobial resistance of selected Salmonella isolates from food animals and food in Alberta

Salmonella isolates (n = 209) obtained from food animals and foods in Alberta during 1996 through 1999 were tested for sensitivity to 17 antimicrobials. Of the 3553 antimicrobial susceptibility tests on Salmonella isolates, 11.8% were positive for resistance. These isolates were commonly resistant to tetracycline (35.4%), streptomycin (32.5%), sulfamethoxazole (28.7%), ticarcillin (27.3%), and ampicillin (26.8%). Resistance to at least 1 antimicrobial was observed in 112 isolates (53.6%). Salmonella Typhimurium, S. Typhimurium var. Copenhagen, and S. Heidelberg were the most common serovars among isolates resistant to individual antimicrobials and multiple antimicrobials. The most common profile of multiple-antimicrobial resistance was that which included resistance to ampicillin, chloramphenicol, streptomycin, sulfamethoxazole, tetracycline, and ticarcillin. The proportions of isolates that were resistant to antimicrobials were greater among bovine isolates of Salmonella than among poultry isolates, and this difference was greater among isolates from veterinary diagnostic sources than among those from monitoring sources.     

Antimicrobial drug resistance and the occurrence of plasmids in Haemophilus paragallinarum

A broth microdilution method was used to examine the sensitivity of 75 Haemophilus paragallinarum isolates to six antimicrobial drugs (ampicillin, erythromycin, neomycin, penicillin, streptomycin, and tetracycline). A total of 55 of the isolates (73%) were sensitive to all six drugs. The remaining 20 isolates were resistant to streptomycin, with one of these isolates also being resistant to tetracycline and another also resistant to neomycin. No isolate showing antimicrobial drug resistance belonged to agglutinin serovar C, despite this being the single largest serovar (27 out of 75) in the study. No plasmids were detected among the 75 isolates, despite the use of five different plasmid-screening techniques.     

Associations between antimicrobial use and the prevalence of antimicrobial resistance in fecal Escherichia coli from feedlot cattle in western Canada

A randomized, controlled, blinded clinical trial was performed at a research feedlot in western Canada. Auction-market-derived steers (n = 288) were randomly assigned to 1 of 3 treatments: 1) no antimicrobials on arrival; 2) oxytetracycline in the starter ration for 14 d; and 3) long-acting oxytetracycline subcutaneously on day 0. Minimal inhibitory concentrations of 7 antimicrobials were determined for 3 generic fecal E. coli isolates per animal on arrival and throughout the feeding period. There was a low prevalence of antimicrobial resistance in generic E. coli isolates from calves on arrival. There were increased proportions of cattle with resistant E. coli isolates early in the feeding period among calves in groups 2 and 3. Individual animal treatments were not associated with increased proportions of cattle with resistant E. coli isolates preslaughter. There was no difference in the proportion of animals with E. coli isolates resistant to tetracycline between the treatment groups preslaughter. However, there were significantly more animals with tetracycline resistant isolates of E. coli preslaughter than at arrival.     

Antibiotic-resistant Escherichia coli isolated from chilled meat at retail outlets

Antibiotic resistance was investigated in Escherichia Coli isolated from beef, veal, lamb and pork at retail level. A total of 100 samples from each meat species was examined. About 16% of the 400 samples were contaminated with resistant E.coli. Significantly more E.coli isolates from pork were drug-resistant than isolates from other meats (P<0.01). About 7% of the combined beef, veal, and lamb E.coli isolates were resistant to two or more antibiotics compared to about 40.0% of the pork isolates (P<0.01) Transfer of resistance was observed for 39.2% of multiple resistant isolates. The results presented form a base for future monitoring of the presence of antibiotic-resistant coliforms on meat suitable for human consumption.     

In vitro antimicrobial sensitivity of isolates of Yersinia pseudotuberculosis from deer

The in vitro sensitivity of 34 isolates of Yersinia pseudotuberculosis from red deer (Cervus elaphus) was determined by disc diffusion tests using 15 antimicrobial drugs. All isolates were sensitive to ampicillin, chloramphenicol, gentamycin, kanamycin, neomycin, “septrin” (combination of trimethoprim and sulphamethoxazole) streptomycin and tetracycline. Twenty isolates were sensitive to polymyxin B and 14 were resistant. All isolates were resistant to bacitracin, erythromycin, novobiocin and moderately resistant to furazolidone. The responses using sulphafurazole or trimethoprim were difficult to interpret accurately but most isolates were classified as sensitive.     

Antimicrobial susceptibility of Escherichia coli isolated from feces of wild cranes migrating to Kagoshima, Japan

Susceptibility to 13 antimicrobial agents was examined for 138 Escherichia coli isolates obtained from 192 fecal samples of wild cranes that migrated for wintering to the Izumi plain, Kagoshima prefecture in Japan. The numbers of isolates that were resistant to the antimicrobials used in this study are as follows: oxytetracycline (OTC), 22 isolates; minocycline, 7 isolates; ampicillin (ABPC), 4 isolates; nalidixic acid, 4 isolates; enrofloxacin, 2 isolates; kanamycin, one isolate. Multidrug resistant isolates exhibiting 2-4 drug resistances were obtained. All of the OTC-resistant isolates carried either the tet (A) or tet(B) gene. The bla(TEM) gene was found in all of the ABPC-resistant isolates.     

Relation of plasmids to virulence and other properties of salmonellae from avian sources

A collection of 185 isolates of 34 serovars of Salmonella from avian sources was examined for plasmids, drug resistance, biochemical properties, serum resistance, and virulence. No serovars other than S. enteritidis, S. typhimurium, and S. heidelberg showed evidence of serovar-associated plasmids. All S. enteritidis isolates carried a single plasmid of 36 Mdal and were resistant to guinea pig serum; one strain that was tested was virulent. Of 27 isolates of S. typhimurium, 11 possessed a 60-Mdal plasmid and 17 harbored a 2.3-Mdal plasmid. Among isolates of S. heidelberg, 21 of 24 carried a 2.2-Mdal plasmid. The only biochemical property that varied was fermentation of inositol, which tended to be related to serovar. Of 172 isolates, 54 were resistant to at least one drug. Multiple drug resistance was usually associated with R plasmids, and transmissible plasmids that encoded resistance to chloramphenicol and gentamicin were demonstrated. Of 117 isolates tested, 43 were resistant to guinea pig serum. Resistance appeared to be a characteristic of isolates rather than serovar and could not be related to plasmids. Twenty-five isolates highly resistant to guinea pig serum were all susceptible to the bactericidal action of chicken serum. In tests for virulence using intraperitoneally (i.p.) and orally inoculated Balb/c mice and day-old chicks, only i.p.-inoculated chicks proved useful in demonstrating large differences among isolates: LD50's ranged from 10(0) to 10(8).     

新疆某猪场不同生长期的猪源大肠杆菌对抗生素耐药性调查

为了调查新疆某规模化养猪场不同生长期的猪源大肠杆菌对临床常用抗生素的耐药情况,从该猪场分别采集育肥猪、妊娠猪、保育猪和哺乳猪的粪样。试验采用微量肉汤稀释法,对从粪样中分离出的大肠杆菌进行最小抑菌浓度测定。结果显示,育肥猪、妊娠猪、保育猪和哺乳猪源大肠杆菌分离率分别为85.2%、83.0%、81.0%和83.0%。不同生长期的猪源大肠杆菌均对氨苄西林和阿莫西林/克拉维酸2种抗生素高度耐药,对头孢噻呋都较为敏感。哺乳猪和保育猪源大肠杆菌耐药情况严重,对多种抗生素高度耐药,以5耐为主;妊娠猪源大肠杆菌轻度耐药,以0耐为主;育肥猪源大肠杆菌以3耐为主。不同生长期的猪源大肠杆菌对临床常用抗菌药物的耐药程度不同。