Detection of Escherichia coli in a cattle manure composting process by selective cultivation and colony polymerase chain reaction

Livestock manure is suitable for use as a composting material. However, various intestinal microbes, such as Escherichia coli, are significant components of such manures. Thus, it is desirable that the level of intestinal microbes, and particularly opportunistic pathogens, in compost is inspected and counted regularly. The sensitivity and specificity of detection of E. coli in compost have been improved by selective cultivation followed by colony polymerase chain reaction (PCR) using the ECO primer. Indeed, the sensitivity of this method is higher than that of DNA extraction from compost and PCR. In this study, changes in numbers of E. coli present in a field-scale composting process over time was assessed using selective cultivation and colony PCR. Numbers of ECO-positive colonies after 24 h decreased, with a concomitant rise in compost temperature. ECO-positive colonies were not detected from 33 to 48 h. However, ECO-positive colony numbers increased beginning on day 4 and continuing until day 42. Thus, it seems likely that the high temperatures reached during the composting process did not affect E. coli numbers in the final compost. Additionally, selective cultivation followed by colony PCR using specific primers is an appropriate method of determining levels of cultivable pathogens in composted materials.     

Antibiotic resistance and genetic diversity of Escherichia coli isolates from traditional and integrated aquaculture in South China

This study investigated antibiotic resistance profiles including antibiotic resistance frequencies, resistance genes and resistance patterns in Escherichia coli strains isolated from traditional and integrated aquaculture systems in South China by using antibiotic susceptibility testing and real time polymerase chain reaction (PCR) technique. The E. coli isolates were found to be resistant to at least one antibiotic among 12 antibiotics. Higher resistance frequencies to ampicillin, sulfamethoxazole, trimethoprime, streptomycin and tetracycline were found compared to the rest antibiotics. Among the 10 tetracycline resistance genes detected in the resistant isolates, the most prevalent tetracycline resistance genes were tetA, tetW and tetB with the frequency of 69.7%, 63.5% and 21.9%, respectively. Three sulfonamide resistance genes were detected in these resistant isolates, with their detection frequencies in the following order: sul2 (55.3%) > sul3 (28.2%) > sul1 (6.2%). Four resistance genes mainly encoding extended-spectrum β-lactamases (ESBLs) were detected in these resistant isolates, with the detection frequencies of blaTEM (28.4%) > blaOXA (9.7%) > blaCTX (9.3%) > blaCARB (5.2%) > blaSHV (0.0%). It was found that the integrated aquaculture system exhibited generally higher prevalence of antibiotic resistance than the traditional aquaculture system. An integrated aquaculture system could facilitate development of bacterial resistance and spread of the antibiotic resistance genes, and consequently become an important reservoir of resistance genes.     

Genotoxicity of methyl parathion in short‐term bacterial test systems

Abstract

Genotoxicity of the insecticide methyl parathion was investigated in Salmonella typhimurium and Escherichia coli bacterial test systems for the detection of back mutations and DNA‐damage. Methyl parathion was mutagenic to S. typhimurium strain TA100 after activation with rat liver microsomal and cytosolic enzymes. In DNA repair tests, methyl parathion was effective in inducing damage to the S. typhimurium strain TA1538 which lack excision repair compared to the strain TA1978 which is proficient in excision repair mechanisms. Normal laboratory light conditions had no effect on the mutagenicity tests, however, exposure of methyl parathion in the petri dish containing the tester strain TA100 and rat liver microsomal and cytosolic enzymes reduced the mutagenic activity and increased the toxic effects of methyl parathion.     

利用基因工程菌BL21处理有机磷混合农药废水的研究

研究了悬浮状态和固定化状态的基因工程菌BL21对有机磷混合农药废水的降解特性.工程菌能快速、高效地降解有机磷混合农药,其最适底物是对硫磷,而马拉硫磷不能被工程菌降解.不同农药降解速率的差别造成了不同有机磷农药的降解过程需要用不同的动力学模型来描述.比较固定化状态和悬浮状态的工程菌的降解效果可知,固定化工程菌的降解活性较后者明显降低,其比降解速率大约仅为后者的20%.考察固定化工程菌长期运行的效果,发现其降解活性保存良好,工程菌稳定性大大提高,未出现固定化细胞溶涨、破碎现象.固定化后,工程菌的比降解速率虽然比悬浮工程菌降低了,但固定化工程菌更适用于长期运行的废水处理系统.     

Quantitative risk analysis for potentially resistant E. coli in surface waters caused by antibiotic use in agricultural systems

Antibiotics are frequently used in agricultural systems to promote livestock health and to control bacterial contaminants. Given the upsurge of the resistant fecal indicator bacteria (FIB) in the surface waters, a novel statistical method namely, microbial risk assessment (MRA) was performed, to evaluate the probability of infection by resistant FIB on populations exposed to recreational waters. Diarrheagenic Escherichia coli, except E. coli O157:H7, were selected for their prevalence in aquatic ecosystem. A comparative study between a typical E. coli pathway and a case scenario aggravated by antibiotic use has been performed via Crystal Ball® software in an effort to analyze a set of available inputs provided by the US institutions including E. coli concentrations in US Great Lakes through using random sampling and probability distributions. Results from forecasting a possible worst-case scenario dose-response, accounted for an approximate 50% chance for 20% of the exposed human populations to be infected by recreational water in the U.S. However, in a typical scenario, there is a 50% chance of infection for only 1% of the exposed human populations. The uncertain variable, E. coli concentration accounted for approximately 92.1% in a typical scenario as the major contributing factor of the dose-response model. Resistant FIB in recreational waters that are exacerbated by a low dose of antibiotic pollutants would increase the adverse health effects in exposed human populations by 10 fold.     

Antimicrobial susceptibility of Escherichia coli isolated from shrimp (Litopenaeus vannamei) and pond environment in northeastern Brazil

This study aimed to test the susceptibility of Escherichia coli strains isolated from the water, bottom sediments and individuals cultivated in shrimp farm ponds, to antibiotics belonging to different families, namely B-Lactams: Imipenem (IPM; 10 μ g), Ampicillin (AMP; 10 μ g), Cephalothin (CEP; 30 μ g), Cefoxitin (FOX; 30 μ g), Ceftriaxone (CRO; 30 μ g); Tetracycline: Tetracycline (TCY; 30 μ g); Aminoglycosides: Gentamicin (GEN; 10 μ g), Amikacin (AMK; 30 μ g); Chloramphenicol: Chloramphenicol (CHO; 30 μ g); Fluoroquinolones: Ciprofloxacin (CIP; 5 μ g); Nitrofurans: Nitrofurantoin (NIT; 300 μ g); Sulfonamides: Trimethoprim-Sulfamethoxazole (SXT; 30 μ g); Quilononas: Nalidixic Acid (NAL; 30 μ g). In the laboratory, the method of dissemination (Test Kirby-Bauer) was performed in order to fulfill the antibiogram tests. The results showed high indices of resistance to Imipenem, Cephalothin and Ampicillin. Chloramphenicol, Nitrofurantoin, Cefoxitin, Ceftiaxone and Ciprofloxacin have displayed the highest index of sensitive strains. The antibiotic resistance index (ARI) and the multiple resistance index (MAR) varied within the ranges of 0.068–0.077 and 0.15–0.39, respectively. More than 90.5% of strains of Escherichia coli showed a variety of resistance profiles to the tested antibiotics. The high indices of resistance may be a consequence of indiscriminate use of antibiotics, but also the transfer of resistance through mobile genetic elements found in shrimp farms.     

Regulatory Oversight of Genetically Engineered Microorganisms: Has Regulation Inhibited Innovation?

/ Using detailed interviews with company representatives and researchers in the field, this paper examines the factors that might account for the slow pace of development of genetically engineered microorganisms (GEMs) intended for environmental release. We specifically analyzed the role of the regulatory system in shaping innovation. We identified at least two cases where industry decided to discontinue the development of a genetically engineered microbial product because of concerns over regulatory oversight. However, most often industry decisions to continue or halt development of GEMs were based on an evaluation of the particular product's efficacy and potential for profitability. Thus the inability of GEMs to perform up to expectations in the field, rather than the regulatory constraints, appears to be the factor responsible for the slow pace of development. KEY WORDS: Genetically engineered microorganisms; Biotechnology; Regulation of biotechnology; Innovation; Environmental release     

Comparative study of fluorogenic and chromogenic media for specific detection of environmental isolates of thermotolerant Escherichia coli

In a field study 78 water samples were analysed employingFluorocult Brilla Broth (BB) and its performance was comparedwith standard MPN procedure. Out of 78 water samples analysed 56(71.7%) samples yielded positive reactions in BB whereas, 50(64.1%) samples were positive by standard fecal coliform test.A comparative study of fluorogenic and chromogenic mediacontaining substrate -D glucuronide for specificdetection of environmental isolates of 313 thermotolerant E.coli has been undertaken. Five fluorogenic media wereused: Fluorocult MacConkey agar (MCA), Fluorocult ECD agar(ECD), Fluorocult VRB agar (VRB), Fluorocult E. coli0157:H7 agar (ECH7) and Fluorocult Brilla Broth (BB) andChromogenic Chromocult agar (CCA). BB and CCA were found to behighly specific and sensitive media to detect E. coli asall E. coli yielded positive reaction on them. On ECH7 andECD agar 67.5 and 64.9 of E. coli isolates gave positivereaction, respectively. Low sensitivity was observed in case ofMCA and VRB agar in detecting E. coli. The performance ofBB appears to be better when compared with standard MPNprocedure employing MacConkey broth/Brilliant green bile brothin detecting E. coli in drinking water.     

孔板与文丘里组合空化杀灭水中病原微生物

利用水力学实验室自主研发的多孔板与文丘里管组合式水力空化装置来杀灭原水中的病原微生物.以水体中大肠杆菌和菌落总数为指示菌,用平板计数法测得不同工况下的大肠杆菌和菌落总数的杀灭率,研究了空化数、孔口排布、孔口数量、运行时间、原水配比浓度、喉部长短对病原微生物杀灭率的影响.试验结果表明：提高孔口流速和喉部流速、延长运行时间、降低空化数、增多孔口数量、改进孔口排布、选取合适配比浓度、延长喉部长度可以提高大肠杆菌和菌落总数的杀灭率.水力空化是一种无消毒副产物、安全、高效的饮用水消毒新技术,具有潜在的应用前景.     

基因工程技术在降解农药中的应用

综述基因工程技术:基因重组、原生质体融合、利用外源基因、降解酶和固相反应器等在降解农药中的研究进展,提出几种高效基因工程降解菌的构建策略和构建手段。