Commercially available garden products as important sources of antibiotic resistance genes—a survey

Environmental Science and Pollution Research - The dissemination of antibiotic resistance genes (ARGs) in the environment contributes to the global rise in antibiotic resistant infections....     

Aerobic inhibition assessment for anaerobic treatment effluent of antibiotic production wastewater

Biological treatment of antibiotic production effluents is an economical approach; however, there are still difficulties to overcome because of the recalcitrant characteristics of these compounds to biodegradation. This study aims to reveal that anaerobic treatment technology can be an option as pretreatment before the activated sludge system treatment to treat antibiotic production effluents. The ISO 8192 method was chosen to test the inhibitory effect of raw and treated antibiotic production effluents in this work. Inhibition tests, which were applied according to ISO 8192, highlighted that the anaerobic treatment effluent is less inhibitory than antibiotic production effluent for activated sludge system. Early EC₅₀ concentrations (30-min values) of raw and treated wastewaters were lower than 180-min values. Also, triple effects (sulfamethoxazole–erythromycin–tetracycline) of antibiotics are more toxic than dual effects (sulfamethoxazole–tetracycline). In light of the experimental results obtained and their evaluation, it can be concluded that anaerobic digestion can be applied as a biological pretreatment method for pharmaceutical industry wastewater including antibiotic mixtures prior to aerobic treatment.     

Relationship between antibiotic residues and occurrence of resistant bacteria in Nile tilapia (Oreochromisniloticus) cultured in cage-farm

The aim of this study was to investigate the relationship between antibiotic residues found in the muscle of cage-farm-raised Nile tilapia (Oreochromisniloticus), the occurrence of resistant bacteria, and the sanitary practices adopted by farmers in Ilha Solteira reservoir, Brazil. Nine fish (three small fish, 40–200 g; three medium-sized fish, 200–500 g; and three large fish, 500–800 g) were collected from four cage farms every three months from April 2013 to January 2014. Ten antibiotic residues were determined using liquid chromatography-mass spectrometry, and bacteria were isolated and tested for antibiotic resistance to determine the multiple antibiotic resistance (MAR) index. Only three antibiotics (oxytetracycline, tetracycline, and florfenicol) were detected in the muscle of Nile tilapia, and their residues were the highest in small fish; however, the MAR index was higher in large fish. In addition, a direct positive relationship between the MAR index and the concentration of antibiotic residues in Nile tilapia was found. Overall, the adoption of prophylactic management practices improved the sanitary status of cage farms, reducing bacterial infections and hampering the development of antibiotic-resistant bacteria.     

Treatment of pharmaceutical wastewater containing antibiotics by O3 and O3/H2O2 processes

Ozonation of three different synthetic pharmaceutical formulation wastewater containing two human antibiotics and a veterinary antibiotic has been studied to enhance the their biodegradability. The effects of pH and initial chemical oxygen demand (COD) value as well as addition of hydrogen peroxide on ozonation process were investigated. Total organic carbon (TOC), COD, biochemical oxygen demand (BOD), and aromatic content (UV254) were the parameters followed to evaluate the performance of ozonation process. Comparison of the biodegradability of selected wastewaters containing different antibiotics confirmed that the variation of biodegradability was associated with the target compound. While BOD5/COD ratio of veterinary antibiotic formulation wastewater was increased from 0.077 to 0.38 with an applied ozone dosage of 2.96 g/l, this ratio for human antibiotic I and human antibiotic II was increased from 0 to 0.1 and 0.27 respectively. Moreover the results of this investigation showed that the ozonation process is capable of achieving high levels of COD and aromaticity removals at about their natural pH values.     

基于红霉素效价当量的不同抗生素生产废水残留效价的测定

为了比较不同废水中残留抗生素及其相关物质的抑菌活性,利用建立的废水残留效价测定方法优选出抑菌活性强、效价线性范围宽的参照抗生素进行抗生素生产废水效价当量测定。以金黄色葡萄球菌（革兰氏阳性菌）作为标准菌株,通过不同抗生素的效价标准曲线从20种抗生素中优选出线性范围宽、抑菌能力强和便宜易得的红霉素作为标准参照抗生素。对土霉素和螺旋霉素2种生产废水的残留效价进行测定,发现用红霉素效价当量与土霉素/螺旋霉素效价当量表征呈显著正相关（p-1,螺旋霉素生产废水1.66 mg·L-1,磺胺甲恶唑生产废水0.56 mg·L-1,6-氨基青霉烷酸生产废水0 mg·L-1）,与几种抗生素本身的抑菌能力的排序一致。     

β-内酰胺类抗生素的环境行为与制药行业源头控制技术研究进展

β-内酰胺类抗生素是目前生产量和使用量最大的抗生素类型之一,在不同的环境基质中都能检出其残留、相关抗药菌和抗性基因,具有潜在的环境风险。中国是世界上最大的β-内酰胺类抗生素原料药生产国,每年产生大量的制药废水和菌渣。如何有效去除制药废水中残留抗生素及效价,并实现制药菌渣资源化利用是行业发展的技术瓶颈。在查阅文献的基础上,梳理了β-内酰胺类抗生素的环境污染来源和污染特征方面的研究进展,阐述了抗生素在环境中的迁移、吸附、水解等环境行为,并从环境抗性的产生和传播角度分析了β-内酰胺类抗生素的环境影响;重点梳理了废水和菌渣中β-内酰胺类抗生素的控制技术方面的研究进展,在此基础上,从降解产物与抗性关系、抗生素和效价的控制目标、废水抗生素和抗性基因控制多级屏障技术体系和制药菌渣无害化及资源化等方面提出展望,以期为环境中β-内酰胺类抗生素的环境管理和风险控制提供参考。     

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.     

The effect of residential and agricultural runoff on the microbiology of a Hawaiian ahupua'a.

The objective of this project was to study the relationship between environmental runoff and the incidence of antibiotic-resistant microorganisms (ARMO) in freshwater streams. Five water systems along the windward coast of the island of O'ahu were evaluated. Samples were collected from sites upstream of residential or agricultural areas, throughout these areas, and at sites of entrance into oceans or bays. It was hypothesized that the incidence of ARMO would increase as the stream received runoff from residential and agricultural areas. The percentage of ARMO did not increase as the streams passed through residential or agricultural areas. Surprisingly, pristine sites, well upstream from residential or agricultural areas, contained bacteria resistant to at least one antibiotic. Areas most affected by runoff did not show a significant increase in the incidence of antibiotic-resistant organisms, suggesting that the incidence of antibiotic resistance is not simply a function of contamination with agricultural or residential runoff. The correlation of antibiotic resistance with heavy metal resistance was evaluated, because others (Fasim et al., 1999; Lazar et al., 2002; Nies, 1999) have shown that antibiotic and heavy metal resistance are each carried on extrachromosomal plasmids. The vast majority of ARMO were also resistant to concentrations of heavy metals reported in the sediments of indicator streams (Waihee, system III), suggesting that an antibiotic-resistant bacterium has a high probability of having dual resistance to a heavy metal. A 3.2-kb plasmid (pSTAMP) was isolated from a bacterium with dual antibiotic and heavy metal resistance. Further analysis of the plasmid is currently in progress.     

Antibiotic resistance,virulence factors and biofilm formation ability in Escherichia coli strains isolated from chicken meat and wildlife in the Czech Republic

Attachment of pathogenic bacteria to food contact surfaces and the subsequent biofilm formation represent a serious threat for the food industry, since these bacteria are more resistant to antimicrobials or possess more virulence factors. The main aim of this study was to investigate the correlation between antibiotic resistance against 13 antibiotics, distribution of 10 virulence factors and biofilm formation in 105 Escherichia coli strains according to their origin. The high prevalence of antibiotic resistance that we have found in wildlife isolates could be acquired by horizontal transfer of resistance genes from human or domestic or farm animals. Consequently, these commensal bacteria might serve as indicator of antimicrobial usage for human and veterinary purposes in the Czech Republic. Further, 46 out of 66 resistant isolates (70%) were able to form biofilm and we found out statistically significant correlation between prevalence of antibiotic resistance and biofilm formation ability. The highest prevalence of antibiotic resistance was observed in weak biofilm producers. Biofilm formation was not statistically associated with any virulence determinant. However, we confirmed the correlation between prevalence of virulence factors and host origin. Chicken isolates possessed more virulence factors (66%), than isolates from wildlife (37%). We can conclude that the potential spread of antibiotic resistance pattern via the food chain is of high concern for public health. Even more, alarming is that E. coli isolates remain pathogenic potential with ability to form biofilm and these bacteria may persist during food processing and consequently lead to greater risks of food contamination.     

Sulfamethoxazole sorption by sediment fractions in comparison to pyrene and bisphenol A

The environmental behavior of antibiotics has attracted great research attention. However, their sorption mechanisms in soils/sediments are still unknown. Comparison of the sorption properties between the widely-studied hydrophobic organic contaminants (HOCs) and antibiotics may provide valuable insight to antibiotic sorption mechanisms. Thus, in this study batch experiments for pyrene (PYR), bisphenol A (BPA), and sulfamethoxazole (SMX) sorption were conducted on a sediment sample and its separated fractions. Our results showed the high sorption of PYR on black carbon and organic matter. Although high sorption of SMX was observed for both separated organic fractions (humic acids) and inorganic mineral particles, the original sediment particles showed relatively low sorption. Competitive sorption between SMX and dissolved humic acid on mineral particles was observed in this study. This competitive interaction is a unique process for antibiotic sorption in soils/sediments compared with apolar HOCs and may be one of the important factors controlling the antibiotic sorption.     

Insights into the effects of Zn exposure on the fate of tylosin resistance genes and dynamics of microbial community during co-composting with tylosin fermentation dregs and swine manure

Environmental Science and Pollution Research - Though heavy metals are widely reported to induce antibiotic resistance propagation, how antibiotic resistance changes in response to heavy metal...     

污泥臭氧氧化处理过程中活菌抗药基因丰度的消减

臭氧氧化技术是一种广泛应用的污泥减量技术,然而臭氧处理能否对污泥中的抗生素抗药基因进行有效消减还不清楚。采用单叠氮溴化丙锭（propidium monoazide,PMA）预处理结合定量PCR（qPCR）方法对污泥臭氧减量过程中不同臭氧消耗量下活性污泥活菌中的四环素、氨基糖苷和大环内酯3大类共21种抗药基因的变化进行了研究。结果表明：臭氧氧化可以有效消减活性污泥活菌中21种抗药基因的绝对丰度（每毫升污泥的抗药基因拷贝数）,在臭氧消耗量0.31 g·g-1（TSS）情况下抗药基因总量降低了75.44%;尽管绝对丰度下降,污泥臭氧氧化处理过程中大量抗药基因的相对丰度（抗药基因拷贝数与细菌16S rRNA基因拷贝数的比例）逐渐增加,表明臭氧处理后污泥中耐药菌占总活菌比例可能增加,具有一定的抗性传播风险;Ⅰ型整合子是抗药基因水平转移的重要遗传元件,其绝对丰度同样随着臭氧消耗量增加而降低,而相对丰度逐渐上升。     

The effect of wastewater treatment on antibiotic resistance in Escherichia coli and Enterococcus sp.

The effects of wastewater treatment on the proportion of Escherichia coli and Enterococcus sp. resistant to specific antibiotics were investigated at two facilities in Davis County, Utah, one of which received hospital waste. Samples were taken from the influent, effluent before disinfection, and secondary anaerobic sludge digester effluent. There was very little difference in antibiotic resistance among E. coli in the inflow waters of the plants but the plant receiving hospital waste had a significantly higher proportion of antibiotic resistant Enterococcus. The effect of wastewater treatment on antibiotic resistance was more pronounced on enterococci than E. coli. Although some increases in antibiotic resistance were observed, the general trend seemed to be a decrease in resistance, especially in the proportion of multidrug resistant Enterococcus sp.     

Tetracycline resistance in semi-arid agricultural soils under long-term swine effluent application

Annually, millions pounds of antibiotics are released unmetabolized into environment along with animal wastes. Accumulation of antibiotics in soils could potentially induce the persistence of antibiotic resistant bacteria. Antibiotics such as tetracyclines and tetracycline-resistant bacteria have been previously detected in fields fertilized with animal manure. However, little is known about the accumulation of tetracyclines and the development of tetracycline resistance in semi-arid soils. Here we demonstrate that continuous land application with swine effluent, containing trace amounts of chlortetracycline, does not necessarily induce tetracycline resistance in soil bacteria. Based on the testing of more than 3,000 bacteria isolated from the amended soils, we found no significant increase in the occurrence and level of chlortetracycline resistant bacteria in soils after 15 years of continuous swine effluent fertilization. To account for a possible transfer of tetracycline-resistant bacteria originated from the swine effluent to soils, we analyzed two commonly found tetracycline resistant genes, tet(O) and tet(M), in the swine effluent and fertilized soils. Both genes were present in the swine effluent, however, they were not detectable in soils applied with swine effluent. Our data demonstrate that agronomic application of manure from antibiotic treated swine effluent does not necessarily result in the development of antibiotic bacterial resistance in soils. Apparently, concentrations of chlortetracycline present in manure are not significant enough to induce the development of antibiotic bacterial resistance.     

Environmental pollution by antibiotics and by antibiotic resistance determinants

Antibiotics are among the most successful drugs used for human therapy. However, since they can challenge microbial populations, they must be considered as important pollutants as well. Besides being used for human therapy, antibiotics are extensively used for animal farming and for agricultural purposes. Residues from human environments and from farms may contain antibiotics and antibiotic resistance genes that can contaminate natural environments. The clearest consequence of antibiotic release in natural environments is the selection of resistant bacteria. The same resistance genes found at clinical settings are currently disseminated among pristine ecosystems without any record of antibiotic contamination. Nevertheless, the effect of antibiotics on the biosphere is wider than this and can impact the structure and activity of environmental microbiota. Along the article, we review the impact that pollution by antibiotics or by antibiotic resistance genes may have for both human health and for the evolution of environmental microbial populations.     

Antimicrobial potential of the ionophore monensin on freshwater biofilm bacteria

Microorganisms play key roles in stream ecosystems, but comparatively little is known about the resilience of freshwater bacterial communities and their susceptibility to the chemical by-products of agricultural land use. Antibiotics used in the agricultural sector are of particular concern and have been detected in waterways associated with agricultural land. Despite widespread agricultural intensification globally and the sector's high antibiotic use, the effects of agricultural antibiotic by-products on stream microbial communities have yet to be characterised. We investigated the impacts of the antibiotic monensin on microbial biofilm communities in a simulated contamination event using streamside-replicated channels. A 24-h pulse experiment in flow channels precolonised by stream biofilm microbial communities contrasted the effects of monensin concentrations ranging from realistic to extreme toxicity levels (1–550 ug L^?1). Biofilm community composition was characterised immediately before and after the pulse for several weeks using automated ribosomal intergenic spacer analysis. Despite applying acutely toxic levels of monensin, only limited effects to biofilm community composition were detected immediately after antibiotic application, and these disappeared within 4 days. Rather, temporal factors drove biofilm differences, highlighting the overriding importance of wider, catchment-level, physiochemical hydrological influences on structuring freshwater biofilm communities, as opposed to localised and sporadic agricultural surface runoff contamination events containing antibiotics.     

林可霉素制药废水的臭氧氧化处理

考察了臭氧氧化对林可霉素的效价削减效果。在初始抗生素浓度为100 mg·L⁻¹时,林可霉素效价削减50%所需消耗的臭氧量为0.118 mg·mg⁻¹抗生素,降解过程符合一级降解动力学特征。进一步采用林可霉素实际废水考察了污水化学需氧量(COD)和pH对抗生素臭氧氧化处理的影响,发现废水的COD每增加100 mg·L⁻¹,则单位抗生素实现50%削减需要增加的臭氧量约为1.64 mg。碱性条件下,臭氧可催化分解生成羟基自由基等活性基团而加速林可霉素的降解。同时,臭氧氧化后林可霉素生产废水的厌氧可生化性提高了98.51%。研究结果可以为林可霉素生产废水的处理技术选择提供参考。     

Antibiotics pollution in Jiulong River estuary: source, distribution and bacterial resistance

To gain insight into the antibiotic pollution in the Jiulong River estuary and the pollutant sources, we analyzed the concentration of 22 widely-used antibiotics in water samples collected from the river and estuary, 17 and 18 sampling sites, respectively. Contamination with sulfonamides, quinolones and chloramphenicols was frequently detected and the distribution pattern of antibiotics suggested that most of the pollutants are from the Jiulong River, especially from the downstream watersheds. To reveal the ecological effects, we isolated 35 bacterial strains from the estuary and analyzed their antibiotic resistance to the eight most frequently detected antibiotics. The bacteria were subsequently classified into seven different genera by 16SrDNA sequencing. Up to 97.1% of the bacteria showed resistance and 70.6% of strains showed multi-resistance to these antibiotics, especially to sulfonamides. This study demonstrated a pattern of antibiotic contamination in the Jiulong River and its estuary and illustrated high bacterial antibiotic resistance which was significantly correlated with the average antibiotics concentrations and detected frequencies in the estuary.     

Fully-automated SPE coupled to UHPLC-MS/MS method for multiresidue analysis of 26 trace antibiotics in environmental waters: SPE optimization and method validation

Environmental Science and Pollution Research - Achieving simultaneous determination of antibiotic multiresidues in environmental waters by solid phase extraction (SPE) coupled with...     

纳米金属对污水处理系统中磺胺耐药基因和耐药菌的影响及其机理

细菌耐药性和纳米材料污染为新兴环境问题。细菌耐药性遗传载体——耐药基因 (ARGs) 广泛存在于污水处理系统中,且已被证明与金属纳米材料密切相关。采用实时荧光定量PCR技术,探究了纳米零价铁 (nZVI) 和纳米氧化锌 (nZnO) 对污水处理系统中磺胺类耐药菌 (ARB) 和ARGs的分布特征的影响。结果表明,50 mg·L⁻¹ nZVI和nZnO暴露均有利于削减磺胺ARB浓度数。与对照组相比,nZVI和nZnO暴露后污泥中sul1丰度增加了0.25%~16.21%,而sul2出现明显削减 (1.51%~15.47%) 。此外,50 mg·L⁻¹ nZVI会导致游离态胞外ARGs大幅削减。nZVI和nZnO暴露通过富集污水处理系统中的intI1、改变细胞膜通透性和调节细菌转录能力促进sul1的增殖。本研究结果阐明了典型纳米金属对污水处理系统中ARB和不同形态ARGs消长的影响及其机制,可为制定有效调控和全面削减污水处理系统中耐药污染策略提供参考。