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1.
An analytical procedure, based on the concept that exposure of bacteria to antibiotics will result in the selection of a resistant population, was developed. Two strains of enteric bacteria, Escherichia coli CS-1 and Enterobacter cloacae B520, which are sensitive to a wide variety of antibiotics, were used as the test organisms. E. coli CS-1 were exposed to 1.00 micrograms antibiotic or antimicrobial/mL; E. cloacae B520 were exposed to 0.01, 0.10, 0.50, 1.00, and 5.00 micrograms/mL. Both organisms developed increased resistance to other antibiotics after exposure to chlortetracycline and oxytetracycline, as measured by the minimum inhibitory concentration (MIC). E. cloacae B520 showed increased resistance to ampicillin, oxytetracycline, and chloramphenicol after exposure to levels as low as 0.10 microgram/mL. Exposure to streptomycin, sulfamethazine, tylosin, bacitracin, flavomycin, virginiamycin, and monensin at levels of 1.00 microgram/mL did not increase the MIC. Exposure to 5.00 micrograms streptomycin, sulfamethazine, tylosin, and monensin/mL increased the MIC of E. cloacae to one of the antibiotic markers. These increased MICs exceeded the 95% confidence limits of the MIC values of the unexposed organisms.  相似文献   

2.
为研究牛粪厌氧发酵沼渣中多重耐药细菌的环境污染风险，选用畜禽养殖中常用的5种抗生素(红霉素、氨苄青霉素、金霉素、链霉素、环丙沙星)对牛粪沼渣中可培养抗生素抗性细菌进行筛选。结果表明：多重耐药细菌的比例高达76.5%,其中，抗5种和4种抗生素的细菌分别有11,21株。所有多重耐药细菌均对氨苄青霉素具有抗性，抗红霉素、金霉素和链霉素的细菌分别占总多重耐药细菌数的92.0%,89.3%,61.3%。通过细菌16S rRNA测序鉴定，32株具有4种以上抗性的细菌分别属于福氏志贺氏菌、摩根氏菌和假中间苍白杆菌，均为重要的临床致病菌。使用全基因组测序对7株典型多重耐药细菌携带的抗性基因进行分析，共检测出28种抗生素抗性基因，对应9种抗生素抗性类型。通过分析抗性基因及插入序列所在位点信息发现，多重耐药细菌普遍携带含有抗性基因和插入序列共存的质粒，表明抗性基因具有高度的可移动性和较强的传播风险。综上所述，牛粪沼渣中含有大量的多重耐药细菌，可视作抗生素抗性基因的储存库，应重视沼渣农业资源化利用过程中的环境风险监测与评估。  相似文献   

3.
Results are compared for the microbiological analysis of antibiotics in feeds using the AOAC plate diffusion assay and the simplified 2-plate assay. Five antibiotics, bacitracin, chlortetracycline, oxytetracycline, penicillin, and streptomycin, were used to supplement feed extracts at levels of 100 and 25 micrograms antibiotic/g feed (bacitracin at 100 micrograms/g only). For bacitracin at the one level and for penicillin at both levels, the 2-plate design yielded significantly more accurate results than those of the AOAC assay. The same was true for the 25 micrograms/g level of oxytetracycline and the 100 micrograms/g level of streptomycin. For streptomycin at the 25 micrograms/g supplementation, the AOAC assay results showed better accuracy. There was no significant difference in results between the 2 designs for oxytetracycline at 100 micrograms/g and chlortetracycline at either level. The accuracy and precision of the results for the 2-plate design are equivalent to or better than those obtained using the AOAC design; in addition, the 2-plate assay is less labor-intensive, is more cost-effective, and is able to determine reasonable conditions of similarity.  相似文献   

4.
以pSET152和pHL212为出发质粒，通过供体大肠杆菌ET12567（pUZ8002）和S17-1属间接合转移肉桂地链霉菌（Streptomyces cinnamonensis），首次构建并优化了肉桂地链霉菌的接合转移系统。利用PCR介导的基因置换技术快速构建了肉桂地链霉菌nsdA基因 （negative regulator of Streptomyces differentiation）中断载体，通过接合转移导入肉桂地链霉菌工业菌株BIB2005，筛选得到一株遗传稳定表型为AmR、KmS的接合子BIB309。PCR分析结果显示该接合子即为nsdA中断突变株。与出发工业菌株相比，在摇瓶水平上nsdA中断突变株BIB309莫能菌素产能提高了2.7倍。  相似文献   

5.
为明确不同抗生素及其处理方式对绿色型豌豆蚜生物学特性的影响,将盐酸金霉素(chlortetracycline HCl)、氯霉素(chloraomycetin)、盐酸土霉素(oxytetracycline HCl)、青霉素G钾盐(penicillin-G K salt)和硫酸链霉素(streptomycin sulfate)5种抗生素分别按同时喷洒蚕豆植株和蚜虫虫体、只喷洒蚜虫虫体和只喷洒蚕豆植株3种方式处理,了解不同处理下豌豆蚜的发育历期、平均体重、体质量差、相对日均体质量增长率和平均产蚜量等生物学参数变化特征。结果表明:3种处理方式对绿色型豌豆蚜的生物学参数影响大小顺序为:同时喷洒植株和虫体处理喷洒植株处理喷洒虫体处理。5种抗生素对绿色型豌豆蚜的生物学参数影响大小顺序为:盐酸土霉素盐酸金霉素硫酸链霉素氯霉素青霉素G钾盐。经过喷洒蚜虫虫体和植株表面联合作用处理,土霉素对绿色型豌豆蚜的生长发育影响最大,若虫期延长2.25 d,整个世代周期延长3.70 d,体重减轻52.37%,体质量差减小55.84%,相对日均体质量增长率减小53.85%,产蚜量下降79.07%;金霉素处理表现为延长发育历期,青霉素、氯霉素和链霉素为缩短发育历期;经5种抗生素处理后的绿色型豌豆蚜体重均减轻,产蚜量下降。土霉素处理对绿色型豌豆蚜的若虫期、世代历期和相对日均体质量增长率的影响与其他4种抗生素差异显著(P0.05),土霉素和金霉素对平均体重和体重差的影响与其他3种抗生素差异显著(P0.05),但相互间差异不显著(P0.05)。经过喷洒植株表面的间接作用处理,土霉素对绿色型豌豆蚜的生长发育影响最大,若虫期龄期延长1.63 d,世代历期延长3.38 d,体重减轻50.28%,体质量差减小51.49%,相对日均体质量增长率减小41.67%,产蚜量下降75.45%;金霉素的影响作用次之,表现为延长发育历期;青霉素、氯霉素和链霉素为缩短发育历期。5种抗生素处理后绿色型豌豆蚜均体重减轻,产蚜量下降。土霉素处理对绿色型豌豆蚜的若虫期、平均体重和产蚜量的影响与其他4种抗生素差异显著(P0.05),土霉素和金霉素对世代历期和相对日均体质量增长率影响差异显著(P0.05),但相互间差异不显著(P0.05)。经过直接喷洒蚜虫虫体间接作用方式处理,土霉素对绿色型豌豆蚜的生长发育影响最大,若虫期延长0.34 d,体重减轻24.32%,相对日均体质量增长率减小26.32%,产蚜量下降44.23%,其他4种抗生素对绿色型豌豆蚜的生物学参数影响较小。土霉素处理对相对日均体质量增长率的影响与其他4种抗生素差异显著(P0.05),土霉素和金霉素对若虫期和产蚜量的影响与其他3种抗生素差异显著(P0.05),但相互间差异不显著(P0.05)。由不同抗生素的不同处理方式进行多重比较得知,对绿色型豌豆蚜生长发育影响较大的3种处理组合为:土霉素同时喷洒植株和虫体处理组合土霉素喷洒植株处理组合金霉素同时喷洒植株和虫体处理组合,3种作用处理间对绿色型豌豆蚜生物学特性影响差异不显著(P0.05),土霉素同时喷洒植株和虫体处理组合对生物学特性的影响与除土霉素喷洒植株处理组合和金霉素同时喷洒植株和虫体组合之外的其他组合处理差异显著(P0.05)。  相似文献   

6.
The presence of antibiotic resistant bacteria was found among standard plate count (SPC) populations of chlorinated drinking water from two districts in Cairo. SPC values obtained were low, ranging between 20 and 3 800 mL?1. Most strains appeared to be ampicillin resistant (89.7%). Those were followed by sulfaguanidine (78.1%) and streptomycin (56.9%). The majority of the tested strains were resistant to two or more antibiotics (multiple antibiotic resistant, MAR) which represent 62.4 to 98 % of the total isolates of the mean obtained in a month. Identification of 363 MAR strains revealed that gram-positive rods were dominant in chlorinated drinking water. Gram-negative fermentative-rods, gram-positive cocci and gramnegative nonfermentative rods represent the second, third and fourth group of the identified MAR phenotypes. The impact of the antibiotic resistant bacteria in drinking water as a health hazard is discussed.  相似文献   

7.
We examined water quality indicators (pH, temperature, turbidity, total phosphorus, and fecal coliform density) and bacterial antibiotic resistance (prevalence, conjugative transfer, and genetic linkage of resistance elements) at locations impacted by confined animal feeding operations (CAFOs) and compared them to nearby reference sites. Sites located upstream and downstream of two wastewater treatment facilities were also compared. Sites near CAFO farms had poor water quality (elevated total phosphorus and turbidity), while water quality remained relatively good downstream of wastewater treatment plants. High proportions of antibiotic-resistant bacteria were observed at all study sites, and frequent conjugative transfer of resistance was observed in laboratory assays. Out of a total of 830 environmental bacterial isolates, 77.1% were resistant to only ampicillin, while 21.2% were resistant to combinations of antibiotics including ampicillin (A), kanamycin (K), chlorotetracycline (C), oxytetracycline (O), and streptomycin (S). Multi-drug-resistant bacteria were significantly more common at sites impacted by CAFO farms. In conjugation assays, 83.3% of the environmental isolates transferred one or more antibiotic resistance genes to a laboratory strain of Salmonella typhimurium. A subset of multi-drug-resistant (A, C, and O) isolates was screened for specific tetracycline resistance genes and class I and II integrons. None of the screened isolates (n?=?22) were positive for integrons, while 13 isolates contained resistance genes for tet (B) and tet (C). Our results indicate that CAFO farms not only impair traditional measures of water quality but may also increase the prevalence of multi-drug-resistant bacteria in natural waters.  相似文献   

8.
A liquid chromatographic (LC) method is described for determination of ampicillin residues in fish tissues. The drug is extracted from tissues with methanol, and the extract is evaporated to dryness. This residue is cleaned up by Florisil cartridge chromatography. LC analysis is carried out on a Nucleosil C18 column, and ampicillin is quantitated by ultraviolet detection at 222 nm. Recoveries of ampicillin added to tissues at levels of 0.2 and 0.1 ppm were 73.2 and 61.5%, respectively. The detection limit was 3 ng for ampicillin standard, and 0.03 ppm in tissues.  相似文献   

9.
The Bacillus stearothermophilus disk assay for penicillin in milk (AOAC official method) was adapted for the determination of ampicillin in fish muscle. The method was evaluated in 2 species of cultured fish: channel catfish and striped bass. Recoveries of ampicillin ranged from 99 to 104% when muscle specimens from both species were spiked at concentrations of 0.025-1.00 micrograms/g. The lower limit of determination (LOD) was 0.025 micrograms/g. The assay was applied to monitor the elimination of ampicillin from the muscle of striped bass after intravascular administration (dosage of 10 mg/kg body weight). The mean concentrations in the muscle declined from 1.160 micrograms/g at 2 h to 0.063 micrograms/g at 18 h. The half-life of ampicillin in the muscle was 3.6 h. Ampicillin concentrations were below LOD at 24 h. No inhibitory activity was observed in the muscle of control fish.  相似文献   

10.
 A large collection of cowpea rhizobia strains was obtained from soil samples collected from either a semi-arid or a tropical rain forest area located at about the same latitude in the north-eastern region of Brazil and evaluated for their intrinsic antibiotic resistance to eight commercial antibiotics. The aim of this study was to correlate antibiotic resistance of native rhizobia strains to edaphic-climatic factors as a way to establish suitable inoculants for specific areas. A large diversity regarding intrinsic antibiotic resistance was found, and 17 clusters were identified as varying from sensitive to gradually resistant up to 500 μg·ml–1 of the antibiotics tested. Clustering analysis did not show any pattern related to the geographic region where isolates have been obtained. On the other hand, an increase in the antibiotic-resistant rhizobia population was associated with an increase in soil P and Al contents. lsolates which were sensitive to spectinomycin, ampicillin, streptomycin, chloramphenicol and tetracycline were present at higher rates in soils devoid of Al. Rhizobia bacteria producing mucus type I (fluid and capable of spreading over the solid media) were found preferentially in soil with Al concentrations up to 36 mg·kg–1, diminishing quickly at higher levels. Received: 2 May 1997  相似文献   

11.
A recently developed confirmatory LC-MS method has been applied to the quantification of five major beta-lactam antibiotics in suspect raw bovine milk samples that gave a positive response with receptor-based (BetaStar) and rapid microbial inhibitory screen tests (Delvotest SP). In total, 18 presumptive positive raw milk samples were reanalyzed; 16 samples showed traces of antibiotic residues that could be identified and quantified by the LC-MS method, ranging from the limits of confirmation up to 38 microg/kg. Of the positive samples, only five (approximately 30%) were found to be violative of EU maximum residue limits. The most frequently detected antibiotic residues were cloxacillin and penicillin G, the former often in combination with amoxicillin or ampicillin. This study compares the results obtained by the three methods on identical samples and addresses how these relate to certain criteria such as sensitivity and selectivity. Furthermore, the limitations of the LC-MS method and the potential impact of the presence of frequently more than one residue in the same milk sample on the response of the rapid test methods are discussed.  相似文献   

12.
O-Methylation, commonly found in synthesis of secondary metabolites of plants and micro-organisms, appears to transfer a methyl group to the hydroxyl group of the recipient which increases the hydrophobicity of the recipient. O-Methyltransferase (OMT), , was isolated and characterized from Streptomyces avermitilis MA-4680. Its amino acid sequence showed 68% similarity with antibiotic C-1027 OMT and 53% similarity with the carminomycin 4-OMT. was expressed in E. coli as a His-tag fusion protein and showed that the methyl was transferred onto the 7-hydroxyl group of the isoflavones, daidzein and genistein, and the flavones, kaempferol and quercetin, as well as the flavanone naringenin. NMR and liquid chromatography-mass spectrometry were used to confirm the location of the methyl group on the recipient compound of naringenin, which was biotransformed into sakuranetin by E. coli transformant expressing (E. coli Sa-2). Therefore, E. coli Sa-2 would be used for the synthesis of the antifungal flavonoid, sakuranetin, through biotransformation.  相似文献   

13.
This study assesses the impact of antibiotics used in clinical and veterinary practices, on the incidence of antibiotic- heavy metal resistant enterics in fresh water and sediment from agricultural and harbor sample sites. A total of 848 bacterial strains of the familyEnterobacteriaceae was isolated from agricultural and lake harbor samples and identified to genus level. These were examined for antibiotic-heavy metal resistance. A select smaller number of these isolates were also examined for the presence of plasmids and ability to transfer antibiotic resistance via conjugation or transformation. More than 85 % of the 848 isolates from all four sites were resistant to Ph, Zn, and Co while 5.6% to 16% were resistant to Te and 2.4% to 5.7% to Hg. The ranking of metal resistance according to frequency was the same as the frequency of metal occurrence in sediment samples. Of the total isolates tested, 87% were resistant to six or more antibiotics and 74% were also simultaneously resistant to Co, Zn, and Ph. Testing the resistance of the water isolates to antibiotics used solely in animal husbandry-veterinary medicine indicated that 55.6% of the agricultural isolates possessed resistance to these antibiotics while only 31.9% of the isolates from harbor water showed resistance to the same antibiotics. Of 41 ampicillin resistant isolates examined, 16 (39%)were capable of transferring antibiotic-heavy resistance markers via conjugation. From this same group, plasmid DNA preparations were made. Of these latter preparations, 67% transformed recipientE. coli cells while 58% possessed discernible, often multiple plasmids when examined by gel electrophoresis.  相似文献   

14.
Isolates of Bradyrhizobium japonicum. native to soils of South Carolina, were obtained from the nodules of eight soybean cultivars grown in field soils with no history of inoculation with antibiotic-resistant bradyrhizobia. A 3-year study screened the native bradyrhizobial populations occupying 6880 nodules for intrinsic resistance to five antibiotic combinations. The sequence of resistance to the antibiotics by the wild-type bradyrhizobia was streptomycin> streptomycin + neomycin> streptomycin + rifampicin> kanamycin + RIFAMPICIN = kanamycin + nalidixic acid. Hence, intrinsic resistance to several antibiotics was the norm for B. japonicum isolated from South Carolina soils.  相似文献   

15.
兽药抗生素的残留状况与环境行为   总被引：1，自引：0，他引：1  
随着畜牧养殖业的迅速发展,抗生素作为人工投入的生产要素得到广泛应用,在带来增产效应的同时,也造成了抗生素的残留,不仅直接影响动物产品的安全与卫生,而且极大的威胁着环境质量安全、人类的身体健康以及生存环境。本文介绍了兽药抗生素的应用及残留状况,概述了兽药抗生素在环境中的迁移、降解及其对动物、植物、微生物的影响等环境行为,并结合国内外研究现状提出了近期兽药抗生素残留研究的重点内容。  相似文献   

16.
Two hypotheses that antibiotic-resistant nodule isolates from uninoculated soybeans grown in a strongly acidic soil were naturally occurring rhizobia which had acquired resistance to spectinomycin and streptomycin or were contaminants from adjacent, inoculated treatments, were tested in laboratory experiments. Soybean nodule isolates (166) as well as 48 cowpea and 89 Rhizobium japonicum strains were used in tests of resistance to six concentrations (0–500 μg ml?1) of kanamycin, spectinomycin and streptomycin, tolerance of stresses of pH 4.6, with or without 50 μm Al, and serological cross-reactivity.More strains from the strongly acidic soil were resistant to the antibiotics than from slightly acidic soils, but resistance to antibiotics did not necessarily entail resistance to pH 4.6 or to 50 μm Al. Twenty-three nodule isolates which were resistant to spectinomycin or streptomycin cross-reacted with antisera of the inoculum strains, indicating that they were contaminants. None of 59 antibiotic-sensitive nodule isolates from uninoculated plants and none of 31 from inoculated plants cross-reacted with the antisera. All 53 antibiotic-resistant isolates from nodules of plants in inoculated plots cross-reacted with test antisera, indicating stability of the antibiotic markers.Cowpea rhizobia were generally more resistant to the antibiotics and more tolerant of pH 4.6 and 50 μm Al than were R. japonicum. Among strains of R. japonicum, slow growers were more resistant to antibiotics than moderately fast growers.  相似文献   

17.
Tylosin, an antibiotic developed specifically for agricultural use, and erythromycin are the main macrolide antibiotics used in animal production. Two-dimensional thin layer chromatography has been used for detection of tylosin in poultry meat, eggs, and milk and for erythromycin in poultry meat. Detection limits reported are, for tylosin, 0.1 ppm in poultry meat, 0.05 ppm in egg, and 0.01 ppm in milk, and for erythromycin, 0.25 ppm in poultry meat. Liquid chromatography (LC) has also been used for determination of tylosin in milk, blood, and tissues of animals. Samples (milk, blood serum, or tissue homogenates in water or pH 2.2 buffer) were deproteinized with acetonitrile, tylosin was partitioned into methylene chloride, and the extracts were concentrated and dissolved in acetonitrile. Chromatography was done on a reverse phase end-capped C18 column using 0.002-0.005 M ammonium dihydrogen phosphate-acetonitrile-methanol (10 + 60 + 30-5 + 80 + 15). Solvent composition was varied with the type of sample analyzed. The method will detect 0.1 ppm tylosin in tissues and less in milk and blood serum. The LC method was more sensitive than microbiological assays for detection of tylosin in tissues of treated swine; recoveries of tylosin by the LC method were frequently several-fold higher.  相似文献   

18.
 Genetic interactions among bacteria are mediated by one of the three distinct gene-exchange mechanisms: conjugation, transformation or transduction. Conjugative gene exchange relies on mobile elements, such as plasmids, which transfer between donor and recipient cells. In natural transformation, competent cells take up DNA and incorporate it into their genome. Gene transfer via transduction is mediated by bacteriophages which accidentally package donor DNA in their phage head and transfer it to recipient cells. Driven mainly by biosafety research and research into the rapid dissemination of antibiotic resistance, the evaluation of gene flux among bacteria in their natural habitats has become a focus of scientific interest in recent years. Accordingly, gene transfer has been assessed in laboratory-based studies employing model ecosystems, as well as in field experiments. Conjugative gene exchange has been shown to occur under a wide range of environmental conditions. Factors identified as conducive for conjugation include the presence of nutrients provided by the rhizosphere of plants. Studies addressing gene transfer via transformation have demonstrated that naturally transformable bacteria develop competence and take up DNA under in situ conditions. Moreover, DNA has been shown to persist to some extent in the environment, and thus be available for uptake by naturally competent cells. Gene exchange via transduction has been demonstrated under conditions of nutrient depletion and low densities of host cells. Whereas gene transfer is readily observed in the laboratory, more importantly, field studies have provided direct evidence that all three gene transfer mechanisms also occur in nature. DNA transfer frequencies observed in the environment in some cases differed considerably from those obtained under laboratory conditions. Transfers of low frequency observed in laboratory-based experiments have been readily detected in the environment in the presence of selective forces. Received: 8 July 1998  相似文献   

19.
Survival of Escherichia coli donor, recipient and transconjugant cells was studied in sterile soil incubated under a variety of conditions. In soil not amended with additional nutrients, the absence of transconjugants indicated that R-plasmid transfer had not occurred. In soil supplemented with a low level of nutrient broth, both donor and recipient cells survived over a 48 hr period. In addition, transconjugant cells were detected in soil as early as two hr after the soil was inoculated with donor and recipient strains.  相似文献   

20.
The use of the selective inhibition (SI) method for measuring fungal:bacterial ratios may be limited due to biocide selectivity and the overlap of antibiotic activity. This study evaluated novel pairs of antibiotics for their specificity in soils of different origins and their potential reduction in inhibition of non-target organisms. Four soils selected for this study were from a semi-arid shrub-steppe, a loblolly pine forest and two grassland sites (restored and farmed prairie plots). Three bactericides were tested: oxytetracycline hydrochloride, streptomycin sulphate, and bronopol. Three fungicides were tested: captan, ketoconazole, and nystatin. The inhibitor additivity ratio and fungal:bacterial ratios were calculated from control and treated soils where inhibition was measured as CO2 respiration reduction with biocides. We were able to minimize non-target inhibition by the antibiotics to <5% and thus calculate reliable fungal:bacterial ratios using captan to inhibit fungi in all four soils, and bronopol to inhibit bacteria in three of the four soils. The most successful bactericide in the restored prairie was oxytetracycline-HCl. Our results demonstrate that application of novel antibiotics is not uniformly successful in soils of different origin and that the SI technique requires more than just optimization of antibiotic concentration; it also requires optimization of antibiotic selection.  相似文献   

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