Multiple antibiotic resistance patterns of Escherichia coli isolates from swine farms.

Antibiotic resistance of Escherichia coli from sows and pigs was determined to compare patterns between pigs of various ages and degrees of antibiotic use. Resistance patterns differed between farm types and pigs of differing ages, indicating that pig age and degree of antibiotic use affect resistance of fecal E. coli.     

Relative resistance to chlorine of poliovirus and coxsackievirus isolates from environmental sources and drinking water

Several poliovirus and coxsackievirus isolates from environmental sources were compared with laboratory strains to determine their rate of inactivation by chlorine. All viruses were tested for up to 1,000 min in the presence of an initial free residual chlorine level of ca. 0.4 mg/liter. Coxsackievirus B5 (CB-5) isolates were found to be more resistant to chlorine than coxsackievirus B4 (CB-4), followed by poliovirus 1, 2, and 3 in order of decreasing resistance to chlorine. Environmental isolates of CB-5 were more resistant than the laboratory strain tested, and for two strains 12 and 22% of the input virus was still infectious after 100 min in the presence of free residual chlorine. Although CB-4 isolates were less resistant to chlorine than CB-5 isolates, after 1,000 min of contact 0.01% of the input virus was still infectious. Except for CB-5 isolates, isolates from environmental sources did not appear to be more resistant to chlorine than laboratory strains. Viruses isolated at different phases during the preparation of drinking water were not more resistant to chlorine and must thus have been protected by other mechanisms.     

Association of metal tolerance with multiple antibiotic resistance of bacteria isolated from drinking water

Bacterial isolates from the drinking water system of an Oregon coastal community were examined to assess the association of metal tolerance with multiple antibiotic resistance. Positive correlations between tolerance to high levels of Cu2+, Pb2+, and Zn2+ and multiple antibiotic resistance were noted among bacteria from distribution waters but not among bacteria from raw waters. Tolerances to higher levels of Al3+ and Sn2+ were demonstrated more often by raw water isolates which were not typically multiple antibiotic resistant. A similar incidence of tolerance to Cd2+ was demonstrated by isolates of both water types and was not associated with multiple antibiotic resistance. These results suggest that simultaneous selection phenomena occurred in distribution water for bacteria which exhibited unique patterns of tolerance to Cu2+, Pb2+, and Zn2+ and antibiotic resistance.     

Relative resistance to chlorine of poliovirus and coxsackievirus isolates from environmental sources and drinking water.

Several poliovirus and coxsackievirus isolates from environmental sources were compared with laboratory strains to determine their rate of inactivation by chlorine. All viruses were tested for up to 1,000 min in the presence of an initial free residual chlorine level of ca. 0.4 mg/liter. Coxsackievirus B5 (CB-5) isolates were found to be more resistant to chlorine than coxsackievirus B4 (CB-4), followed by poliovirus 1, 2, and 3 in order of decreasing resistance to chlorine. Environmental isolates of CB-5 were more resistant than the laboratory strain tested, and for two strains 12 and 22% of the input virus was still infectious after 100 min in the presence of free residual chlorine. Although CB-4 isolates were less resistant to chlorine than CB-5 isolates, after 1,000 min of contact 0.01% of the input virus was still infectious. Except for CB-5 isolates, isolates from environmental sources did not appear to be more resistant to chlorine than laboratory strains. Viruses isolated at different phases during the preparation of drinking water were not more resistant to chlorine and must thus have been protected by other mechanisms.     

Metal tolerance and antibiotic resistance patterns of a bacterial population isolated from sea water

The total aerobic heterotrophic and metal-resistant bacterial communities were studied in marine water. The resistance patterns, expressed as MICs, for 81 bacterial isolates to eight heavy metals were surveyed by using the agar dilution method. A great proportion of the isolates were sensitive to cadmium (99%), mercury (91%), zinc (84%) and cobalt (83%). On the other hand, 94%, 40%, 35% and 22% were resistant to lead, nickel, arsenate and copper, respectively. The majority of the tested strains (95.06%) were multiple metal-resistant, with pentametal resistance as the major pattern (25.9%). The response of the isolates to 11 tested antibiotics was tested and ranged from complete resistance to total sensitivity and multiple antibiotic resistance was exhibited by 70.38% of the total isolated population. The highest incidence of metal-antibiotic double resistance existed between lead and all antibiotics (100%), copper and penicillin (95%) and nickel and ampicillin (83.3%).     

Association of metal tolerance with multiple antibiotic resistance of bacteria isolated from drinking water.

Bacterial isolates from the drinking water system of an Oregon coastal community were examined to assess the association of metal tolerance with multiple antibiotic resistance. Positive correlations between tolerance to high levels of Cu2+, Pb2+, and Zn2+ and multiple antibiotic resistance were noted among bacteria from distribution waters but not among bacteria from raw waters. Tolerances to higher levels of Al3+ and Sn2+ were demonstrated more often by raw water isolates which were not typically multiple antibiotic resistant. A similar incidence of tolerance to Cd2+ was demonstrated by isolates of both water types and was not associated with multiple antibiotic resistance. These results suggest that simultaneous selection phenomena occurred in distribution water for bacteria which exhibited unique patterns of tolerance to Cu2+, Pb2+, and Zn2+ and antibiotic resistance.     

产ESBLs大肠埃希菌和肺炎克雷伯菌呼吸道分离株的基因分型及耐药性分析

目的了解深圳市人民医院大肠埃希菌和肺炎克雷伯菌呼吸道分离株超广谱β-内酰胺酶(ESBLs)的基因型特点及耐药性。方法采用临床实验室标准化协会(CLSI)推荐的表型确证试验筛选出该院呼吸道分离株产ESBLs大肠埃希菌和肺炎克雷伯菌共78株。应用PCR及DNA测序法分析产酶株的TEM、SHV及CTX-M3种β-内酰胺酶基因,用琼脂稀释法测定细菌最低抑菌浓度(MIC)。结果 37株产ESBLs大肠埃希菌中,28株(75.7%)检出CTX-M-14基因,4株(10.8%)检出CTX-M-9基因,其他型较少见。41株肺炎克雷伯菌中,25株(61.0%)检出SHV-12基因,4株(9.8%)检出SHV-11基因,其他SHV型较少。20株(48.8%)检出CTX-M-14基因,5株(12.2%)检出CTX-M-3基因,其他型较少。产ESBL菌株均对亚胺培南敏感,对氨苄西林/舒巴坦的耐药率最高(90%),对其他抗生素有不同程度耐药。结论深圳市人民医院呼吸道分离的产ESBLs大肠埃希菌以CTX-M-14型为主,产酶肺炎克雷伯菌以SHV-12和CTX-M-14型为最常见。     

Analysis of antibiotic susceptibility patterns of Helicobacter pylori isolates from Malaysia

Background: The prevalence of antibiotic resistance varies in geographic areas. The information on the antibiotic susceptibility patterns of Helicobacter pylori (H. pylori) in our local setting is therefore relevant as a guide for the treatment options. Objective: This study was conducted to determine the primary resistance rates among H. pylori isolated from Malaysian patients. Materials and methods: Biopsy samples were obtained from the stomach antrum and corpus of 777 patients from September 2004 until 2007. H. pylori isolated from these patients were then subjected to minimum inhibitory concentration (MICs) determination using E‐test method, against metronidazole, clarithromycin, levofloxacin, ciprofloxacin, amoxicillin, and tetracycline. Results: From 777 patients, 119 were positive for H. pylori where a total of 187 strains were isolated. The resistance rates were noted to be 37.4% (metronidazole), 2.1% (clarithromycin), 1% (levofloxacin and ciprofloxacin), and 0% (amoxicillin and tetracycline). Different resistance profiles were observed among isolates from the antrum and corpus of 13 patients. Resistance to one type of antibiotic was observed in 36.4% of the strains where mono‐resistance to metronidazole was the most common. Resistance to ≥2 antibiotics was noted in 3.3% of isolates. High metronidazole MICs of ≥256 μg/mL were observed among the resistant strains. Conclusions: The resistance rates of the antibiotics used in primary treatment of H. pylori infections in Malaysia are low, and multi‐antibiotic‐resistant strains are uncommon. Infections with mixed populations of metronidazole‐sensitive and ‐resistant strains were also observed. However, the high metronidazole MIC values seen among the metronidazole‐resistant strains are a cause for concern.     

Virulence and antibiotic susceptibility of Aeromonas spp. isolated from drinking water

Aeromonas isolates from tap water, mineral water, and artesian well water were investigated for their ability to produce different potential virulence factors or markers such as hemolysins, cytotoxins, phospholipase, DNase, hydrophobicity and their ability to adhere to epithelial cells and to abiotic surfaces. The susceptibility to antibiotics of Aeromonas isolates was also examined. Majority of the isolates displayed hemolytic activity against sheep erythrocytes, while only 7 of the 23 Aeromonas strains displayed DNase activity and 4 of the 23 Aeromonas strains tested were regarded as positive for phospholipase production. Most of the isolates showed cytotoxic activities in culture filtrate dilutions at titer of 1/8 or lower. No general relation between the strain isolated and the ability to interact with epithelial cells could be established. Using the bacterial adherence to hydrocarbons method, most of the strains were classified as highly hydrophilic. All five Aeromonas jandaei strains isolates, 9 of the 12 Aeromonas sp strains and four of the five Aeromonas hydrophila were multidrug resistant. The most active antimicrobial was ciprofloxacin (susceptible in 100% of the isolates), and the least active antibiotic was ampicillin (resistance in 92% of the isolates). The majority of the isolates tested were not killed by chlorine at 1.2 mg/l. Whether the high tolerance to chlorine of Aeromonas isolates can be linked to greater virulence is not know.     

379株血培养病原菌的临床分布及耐药性分析

目的了解血培养病原菌种类、临床分布及耐药情况。方法对2008年6月至2009年6月的379株血培养病原菌及其药敏情况用WHONET5.4软件进行统计分析。结果 379株病原菌中,分离率从高到低依次是凝固酶阴性葡萄球菌(133/35.1%)、大肠埃希菌(44/11.6%)、肺炎克雷伯菌(32/8.4%)、鲍曼不动杆菌(30/7.9%)、铜绿假单胞菌(25/6.6%)、金黄色葡萄球菌(18/4.7%)、肠球菌(13/3.4%)和其他(75/22.2%)。这些菌株主要分布在ICU和普外科。药敏结果分析显示:革兰阳性球菌中,未发现利奈唑胺和万古霉素耐药株。金黄色葡萄球菌和凝固酶阴性葡萄球菌对苯唑西林的耐药率分别为61.1%和89.5%;屎肠球菌对多数抗生素耐药率超过80%,粪肠球菌对青霉素类抗生素敏感性较高。革兰阴性杆菌中,大肠埃希菌和肺炎克雷伯菌中产ESBLs菌株比例分别为36.4%和31.3%,且发现2株耐亚胺培南的肺炎克雷伯菌;鲍曼不动杆菌除对卡那霉素保持敏感外,对其他抗生素的耐药率为50%~100%;铜绿假单胞菌对头孢曲松的耐药率为100%,对其他β-内酰胺类抗生素、氨基糖苷类和喹诺酮类抗生素的耐药率相对较低。结论临床上应规范血培养标本留取方法以减少污染,加强细菌耐药监测、严格抗生素使用,以延缓细菌耐药情况的发生和发展。     

Characterization of Sphingomonas isolates from Finnish and Swedish drinking water distribution systems

Sphingomonas species were commonly isolated from biofilms in drinking water distribution systems in Finland (three water meters) and Sweden (five water taps in different buildings). The Sphingomonas isolates (n = 38) were characterized by chemotaxonomic, physiological and phylogenetic methods. Fifteen isolates were designated to species Sphingomonas aromaticivorans, seven isolates to S. subterranea, two isolates to S. xenophaga and one isolate to S. stygia. Thirteen isolates represented one or more new species of Sphingomonas. Thirty-three isolates out of 38 grew at 5 degrees C on trypticase soy broth agar (TSBA) and may therefore proliferate in the Nordic drinking water pipeline where the temperature typically ranges from 2 to 12 degrees C. Thirty-three isolates out of 38 grew at 37 degrees C on TSBA and 15 isolates also grew on blood agar at 37 degrees C. Considering the potentially pathogenic features of sphingomonas, their presence in drinking water distribution systems may not be desirable.     

Chlorine resistance of poliovirus isolants recovered from drinking water

Poliovirus 1 isolants were recovered from finished drinking water produced by a modern, well-operated water treatment plant. These waters contained free chlorine residuals in excess of 1 mg/liter. The chlorine inactivation of purified high-titer preparations of two such isolants was compared with the inactivation behavior of two stock strains of poliovirus 1, LSc and Mahoney. The surviving fraction of virus derived from the two natural isolants was shown to be orders of magnitude greater than that of the standard strains. These results raise the question whether indirect drinking water standards based on free chlorine residuals are adequate public health measures, or whether direct standards based on virus determinations might be necessary.     

长沙地区临床分离金黄色葡萄球菌的耐药监测

目的了解长沙地区临床分离金黄色葡萄球菌（以下简称金葡菌）对常用抗菌药物的耐药现状,探讨金黄色葡萄球菌对甲氧西林的耐药水平。方法收集长沙地区11家医院2009年11月至2010年11月临床分离的非重复金葡菌279株,应用Vitek-2全自动微生物分析系统进行鉴定,K-B法检测金葡菌对24种药物的敏感性,产色头孢菌素试验检测β-内酰胺酶以及D试验检测诱导型克林霉素耐药。应用头孢西丁和苯唑西林纸片扩散法筛查耐甲氧西林的金葡菌（MRSA）,琼脂稀释法检测头孢西丁和苯唑西林的最低抑菌浓度（MIC）。结果在被检测的24种药物中,敏感率〉50%的药物为9种,未发现对万古霉素、替考拉宁和利奈唑胺耐药菌株;耐药率〉50%的抗菌药物有11种,其中以青霉素和氨苄西林的耐药率最高（均为97.1%）。MRSA的分离率达54.5%,且对常用的16种抗菌药物的耐药率均显著高于甲氧西林敏感金黄色葡萄球菌（MSSA）。279株金葡菌中,β-内酰胺酶阳性250株（89.6%）;红霉素耐药而克林霉素敏感或中介的30株中,D试验阳性22株（73.3%）。苯唑西林（OXA）和头孢西丁（FOX）MIC范围分别为0.125～〉256μg/mL和2～〉256μg/mL,苯唑西林的MIC50和MIC90分别为128μg/mL和256μg/mL,头孢西丁的MIC50和MIC90分别为64μg/mL和256μg/mL。结论长沙地区临床分离金葡菌对常用抗菌药物呈多重耐药;MRSA不仅分离率高,而且对甲氧西林呈高水平耐药。     

Diversity of species and antibiotic resistance in enterococci isolated from seafood in Tunisia

The purpose of this study was to analyse the antibiotic resistance and virulence of enterococci recovered from seafood and to characterise the associated genes. Forty-four enterococcal isolates [Enterococcus faecalis (21), E. faecium (11), E. casseliflavus (5), E. durans (3), E. hirae (2), E. gallinarum (1) and E. mundtii (1)] were recovered from 70 samples of seafood collected during March–May 2015 in Tunisia. Isolates were tested for antibiotic resistance to 12 antibiotics by the disc diffusion method. Rates of resistance in the range 25–45.5% were observed for pristinamycin, ciprofloxacin, streptomycin, tetracycline and erythromycin, and in the range 6.8–9.1% for kanamycin, gentamicin and chloramphenicol. However, all strains showed susceptibility to β-lactams and glycopeptides. Multi-resistance to at least three different classes of antibiotics was detected in 14 strains (31.8%). Among 12 tetracycline-resistant enterococci, tet(M) was detected in 11 isolates and tet(L) in seven isolates. The erm(B) gene was identified in 91% of erythromycin-resistant isolates. All chloramphenicol-resistant isolates carried the cat gene, and all kanamycin-resistant isolates harboured the aph(3)-IIIa gene. The aac(6′)-aph(2″) and ant(6)-Ia genes were detected in high-level gentamicin- and streptomycin-resistant isolates, respectively. The virulence genes gelE (29.5%), esp (9.1%), cylA and cylB (9.1%) were found in enterococci. This is the first study in Tunisia to underscore the importance of seafood as a reservoir of enterococci carrying resistance and virulence genes.     

Incidence of antibiotic resistance and virulence determinants among Enterococcus italicus isolates from dairy products

The main objective of this work was to investigate the biosafety of Enterococcus italicus, a recently described enterococcal species widely diffused in dairy products. For this purpose, the antibiotic susceptibility and the incidence of virulence factors among 30 E. italicus isolates originating mainly from different Italian cheeses were tested. Although not all 30 isolates showed unique genotypes, PCR fingerprinting evidenced a notable genotypic diversity among the E. italicus collection under study. All isolates were susceptible to vancomycin, gentamicin, erythromycin, ampicillin, chloramphenicol and bacitracin. Five isolates corresponding to three unique genotypes exhibited phenotypic resistance to tetracycline with MICs ranging from 64-256mug/ml. By PCR-based detection, the genetic basis of the Tet(R) phenotype in these strains was linked to the tet(S) gene whereas detection of tet(L) and tet(M) genes and the integrase element int of the Tn916/Tn1545 family of transposons were negative. Likewise, none of the strains appeared to contain any of the tested virulence genes gelE, asaI, cpd, agg, cylA, cylB, cylM, ace and hyl(Efm). The results of this study warrant further research into the environmental dissemination of Tet(R)E. italicus and into the potential transferability of its tet(S) genes.     

Temporal dynamics and impact of manure storage on antibiotic resistance patterns and population structure of Escherichia coli isolates from a commercial swine farm

Many confined-livestock farms store their wastes for several months prior to use as a fertilizer. Storing manure for extended periods could significantly bias the composition of enteric bacterial populations subsequently released into the environment. Here, we compared populations of Escherichia coli isolated from fresh feces and from the manure-holding tank (stored manure) of a commercial swine farm, each sampled monthly for 6 months. The 4,668 confirmed E. coli isolates were evaluated for resistance to amikacin, ampicillin, cephalothin, chloramphenicol, kanamycin, nalidixic acid, streptomycin, sulfamethoxazole, tetracycline, trimethoprim, and trimethoprim plus sulfamethoxazole. A subset of 1,687 isolates was fingerprinted by repetitive extragenic palindromic PCR (rep-PCR) with the BOXA1R primer to evaluate the diversity and the population structure of the collection. The population in the stored manure was generally more diverse than that in the fresh feces. Half of the genotypes detected in the stored manure were never detected in the fresh fecal material, and only 16% were detected only in the fresh feces. But the majority of the isolates (84%) were assigned to the 34% of genotypes shared between the two environments. The structure of the E. coli population showed important monthly variations both in the extent and distribution of the diversity of the observed genotypes. The frequency of detection of resistance to specific antibiotics was not significantly different between the two collections and varied importantly between monthly samples. Resistance to multiple antibiotics was much more temporally dynamic in the fresh feces than in the stored manure. There was no relationship between the distribution of rep-PCR fingerprints and the distribution of antibiotic resistance profiles, suggesting that specific antibiotic resistance determinants were dynamically distributed within the population.     

Antibiotics and antibiotic resistance in water environments

Antibiotic-resistant organisms enter into water environments from human and animal sources. These bacteria are able to spread their genes into water-indigenous microbes, which also contain resistance genes. On the contrary, many antibiotics from industrial origin circulate in water environments, potentially altering microbial ecosystems. Risk assessment protocols for antibiotics and resistant bacteria in water, based on better systems for antibiotics detection and antibiotic-resistance microbial source tracking, are starting to be discussed. Methods to reduce resistant bacterial load in wastewaters, and the amount of antimicrobial agents, in most cases originated in hospitals and farms, include optimization of disinfection procedures and management of wastewater and manure. A policy for preventing mixing human-originated and animal-originated bacteria with environmental organisms seems advisable.     

Diversity and significance of mold species in Norwegian drinking water

In order to determine the occurrence, distribution, and significance of mold species in groundwater- and surface water-derived drinking water in Norway, molds isolated from 273 water samples were identified. Samples of raw water, treated water, and water from private homes and hospital installations were analyzed by incubation of 100-ml membrane-filtered samples on dichloran-18% glycerol agar. The total count (number of CFU per 100 ml) of fungal species and the species diversity within each sample were determined. The identification of mold species was based on morphological and molecular methods. In total, 94 mold species belonging to 30 genera were identified. The mycobiota was dominated by species of Penicillium, Trichoderma, and Aspergillus, with some of them occurring throughout the drinking water system. Several of the same species as isolated from water may have the potential to cause allergic reactions or disease in humans. Other species are common contaminants of food and beverages, and some may cause unwanted changes in the taste or smell of water. The present results indicate that the mycobiota of water should be considered when the microbiological safety and quality of drinking water are assessed. In fact, molds in drinking water should possibly be included in the Norwegian water supply and drinking water regulations.     

Antibiotic resistance in bacterial isolates from laboratory animal colonies naive to antibiotic treatment

Antiobiogrammes were made of a number of isolates of Staphylococcus aureus, Escherichia coli and Pasteurella pneumotropica derived from rodent, rabbit or minipig colonies never treated with antibiotics. For S. aureus no differences between rats and mice were found in the percentage of resistant isolates. Gentamicin and erythromycin were found to be the most efficient, while the highest percentages of resistance were found to be against penicillins and sulphonamides. In general, the results from antibiogrammes on E. coli were rather uniform, with only slight differences between isolates from different species, except that more vancomycin and tetracycline-resistant minipig isolates were found. In almost all isolates of E. coli, resistance was shown against penicillin, fucidin, macrolides, lincosamides and tiamulin. For a number of antibiotics, mouse isolates of P. pneumotropica were more frequently found to be sensitive than rat isolates. The resistance patterns of E. coli from the minipigs were quite similar to resistance patterns found in farm pigs, but apart from this, the resistance patterns of the bacterial species tested did not resemble human or farm animal patterns in any of the animal species, and, therefore, these studies do not support the theory that S. aureus and E. coli in laboratory animal colonies derive from the normal flora of the human caretakers. The fact that rodent species of E. coli, in contrast to human and farm animal species, are sensitive to ampicillin, tetracyclines, and the combination of sulphonamides and trimethoprim, might be due to the fact that these antibiotics are not used in rodent populations.