Sediment samples from three coastal sites - two beach resorts (Beach 1 and Beach 2 sites) and an area lying between an oil refinery and a river estuary (Estuarine site) - were analyzed for antibiotic- and heavy metal (HM)-resistant enterococci.A total of 123 enterococci, 36 E. faecium, 34 E. casseliflavus, 33 E. hirae, 5 E. faecalis, 3 E. durans, 3 E. gallinarum, and 9 Enterococcus spp, were recovered. Strains resistant to erythromycin, tetracycline and quinupristin/dalfopristin (Q/D) were recovered from all sites, whereas multidrug-resistant isolates were recovered only from “Beach 2” (14%) and “Estuarine” (3.7%). As regards HM resistance, the strains showed a high frequency (68%) of cadmium and/or copper resistance and uniform susceptibility to mercury. The prevalence of cadmium-resistant strains was significantly higher among erythromycin-resistant than among erythromycin-susceptible strains. A significant association between cadmium or copper resistance and Q/D resistance was also observed at “Estuarine” site. The levels of the two HMs in sediment from all sites were fairly low, ranging from 0.070 to 0.126 μg/g, for cadmium and from 1.00 to 7.64 μg/g for copper. Mercury was always undetectable. These findings are consistent with reports that low HM concentrations may contribute to co-selection of antibiotic-resistant bacterial strains, including enterococci.

Environmental observations of antibiotics and other pharmaceuticals have received attention as indicators of an urbanizing global water cycle. When connections between environment and development of antibiotic resistance (ABR) are considered, it is increasingly important to understand the life cycle of antibiotics. Here we examined the global occurrence of erythromycin (ERY) in: 1. wastewater effluent, inland waters, drinking water, groundwater, and estuarine and coastal systems; 2. sewage sludge, biosolids and sediments; and 3. tissues of aquatic organisms. We then performed probabilistic environmental hazard assessments to identify probabilities of exceeding the predicted no-effect concentration (PNEC) of 1.0 μg L−1 for promoting ABR, based on previous modeling of minimum inhibitory concentrations and minimal selective concentrations of ERY, and measured levels from different geographic regions. Marked differences were observed among geographic regions and matrices. For example, more information was available for water matrices (312 publications) than solids (97 publications). ERY has primarily been studied in Asia, North America and Europe with the majority of studies performed in China, USA, Spain and the United Kingdom. In surface waters 72.4% of the Asian studies have been performed in China, while 85.4% of the observations from North America were from the USA; Spain represented 41.9% of the European surface water studies. Remarkably, results from PEHAs indicated that the likelihood of exceeding the ERY PNEC for ABR in effluents was markedly high in Asia (33.3%) followed by Europe (20%) and North America (17.8%). Unfortunately, ERY occurrence data is comparatively limited in coastal and marine systems across large geographic regions including Southwest Asia, Eastern Europe, Africa, and Central and South America. Future studies are needed to understand risks of ERY and other antibiotics to human health and the environment, particularly in developing regions where waste management systems and treatment infrastructure are being implemented slower than access to and consumption of pharmaceuticals is occurring.

Excessive use of antibiotics in breeding industry leads to accumulation of antibiotic residuals and antibiotic resistance genes (ARGs) in environment from improperly treated livestock excrements. Four commonly used veterinary antibiotics including chlortetracycline (CTC), sulfamerazine (SMZ), enrofloxacin (ENR) and erythromycin (ERY) were monitored in the swine manure composting. Co-resistance and cross-resistance effects among relative ARGs, correlations between ARGs and bacterial community under multiple antibiotics residual during the composting were investigated in this research. With CTC addition up to 20 mg/kg, more than 99% of CTC removal was achieved after composting, and most of the other antibiotics can be thoroughly removed as well. The variations in ARGs during the composting were strongly correlated to the compositions of the microbial community, Bacteroides and Sporosarcina were main ARGs carriers in the thermophlic phase. Clostridium Ⅺ, Clostridium sensu stricto, and Pseudoxanthomonas, might spread ARGs in cooling and maturing stage. Most of the tested ARGs in swine manure can be effectively reduced through composting, thus makes the compost products safe for soil fertilization.

Erythromycin is an antibiotic employed in the treatment of infections caused by Gram positive microorganisms and the increasing use has made it a contaminant of emerging concern in aqueous ecosystems. Cerium oxide nanoparticles (CeO₂ NPs), which are known to have catalytic and antioxidant properties, have also become contaminants of emerging concern. Due to the high reactivity of CeO₂ NPs, they can interact with erythromycin magnifying their effects or on the other hand, considering the redox potential of CeO₂ NPs, it can alleviate the toxicity of erythromycin. The present study was carried out to assess the toxicity of both single compounds as well as mixed on Chlamydomonas reinhardtii and Phaeodactylum tricornutum (freshwater and marine microalgae respectively) employed as target species in ecotoxicological tests. Mechanisms of oxidative damage and those harmful to the photosynthetic apparatus were studied in order to know the toxic mechanisms of erythromycin and the joint effects with CeO₂ NPs. Results showed that erythromycin inhibited the microalgae population growth and effective quantum yield of PSII (E.Q.Y.) in both microalgae. However, the freshwater microalgae Chlamydomonas reinhardtii was more sensitive than the marine diatom Phaeodactylum tricornutum. Responses related to the photosynthetic apparatus such as E.Q.Y. was affected by the exposure to erythromycin of both microalgae, as chloroplasts are target organelle for this antibiotic.Mixed experiments (CeO₂ NPs + erythromycin) showed the protective role of CeO₂ NPs in both microalgae preventing erythromycin toxicity in toxicological responses such as the growth of the microalgae population and E.Q.Y.

A mesocosm case study was conducted to gain understanding and practical knowledge on biofilm emerging contaminants biodegradation capacity under stressor and multiple stressor conditions. Two real life scenarios: I) biodegradation in a pristine intermittent stream experiencing acute pollution and II) biodegradation in a chronically polluted intermittent stream, were examined via a multifactorial experiment using an artificial stream facility. Stream biofilms were exposed to different water flow conditions i.e. permanent and intermittent water flow. Venlafaxine, a readily biodegradable pharmaceutical was used as a measure of biodegradation capacity while pollution was simulated by a mixture of four emerging contaminants (erythromycin, sulfisoxazole, diclofenac and imidacloprid in addition to venlafaxine) in environmentally relevant concentrations. Biodegradation kinetics monitored via LC-MS/MS was established, statistically evaluated, and used to link biodegradation with stress events. The results suggest that the effects of intermittent flow do not hinder and may even stimulate pristine biofilm biodegradation capacity. Chronic pollution completely reduced biodegradation in permanent water flow experimental treatments while no change in intermittent streams was observed. A combined effect of water flow conditions and emerging contaminants exposure on biodegradation was found. The decrease in biodegradation due to exposure to emerging contaminants is significantly greater in streams with permanent water flow suggesting that the short and medium term biodegradation capacity in intermittent systems may be preserved or even greater than in perennial streams.

Heavy metal contamination of source water frequently occurred in developing countries as a result of accidents. To address the problems, most of the previous studies have focused on engineering countermeasures. In this study, we investigated the effects of heavy metals, particularly copper, on the development of antibiotic resistance by establishing a copper shock loading test. Results revealed that co-selection occurred rapidly within 6 h. Copper, at the levels of 10 and 100 mg/L, significantly increased bacterial resistance to the antibiotics tested, including rifampin, erythromycin, kanamycin, and a few others. A total of 117 antimicrobial-resistance genes were detected from 12 types of genes, and the relative abundance of most genes (particularly mobile genetic elements intⅠand transposons) was markedly enriched by at least one fold. Furthermore, the copper shock loading altered the bacterial community. Numerous heavy metal and antibiotic resistant strains were screened out and enriched. These strains are expected to enhance the overall level of resistance. More noticeably, the majority of the co-selected antibiotic resistance could sustain for at least 20 h in the absence of copper and antimicrobial drugs. Resistance to vancomycin, erythromycin and lincomycin even could remain for 7 days. The prominent selection pressure by the copper shock loading implies that a real accident most likely poses similar impacts on the water environment. An accidental release of heavy metals would not only cause harm to the ecological environment, but also contribute to the development of bacterial antibiotic resistance. Broader concerns should be raised about the biological risks caused by sudden releases of pollutants by accidents.

The ubiquity of pharmaceuticals in the aquatic environment and the accumulation in organisms of lower trophic levels have been documented. The immunotoxicity of these xenobiotics has however been little investigated. This study assessed the effects of pharmaceuticals on the immune responses of harbor seal lymphocytes. Peripheral blood mononuclear cells isolated from harbor seal pups were exposed to varying concentrations of 17α-ethinyl estradiol (250–50,000μg/L), naproxen (500–100,000μg/L), carbamazepine (500–100,000μg/L), erythromycin (750–150,000μg/L) and binary mixtures thereof in vitro. All individual compounds and mixtures inhibited lymphocyte proliferation. Mixture effects were non-additive and predictive values overestimated the inhibition of proliferation. Male pups were more sensitive to erythromycin exposure. Comparison with the sensitivity of the 11B7501 cell line showed a higher sensitivity of pups to individual compounds and the inverse trend for mixtures. Based on our results, we hypothesize that pharmaceuticals may have the potential to interrupt immune functions in harbor seals.

Rivers may receive pharmaceuticals, personal care products, and environment estrogens, which are emerging concerns, from various sources. Understanding the fate of these emerging contaminants (ECs) from the sources to their receiving river is important for assessing their ecosystem risk. Here, the occurrence, seasonal variation, spatial distribution, and ecological risk of 22 ECs in water and sediments from the Jilin Songhua River, as well as in the effluents from the riverside Jilin wastewater treatment plant (WWTP) were investigated. Results indicated that estriol with the highest median concentration of 21.5 ng L⁻¹ in the river water and with the highest median concentration of 481.5 ng g⁻¹ in the sediments, and methylparaben with the highest concentration of 29.6 ± 2.9 ng L⁻¹ in the WWTP effluents were the predominant contaminants. The total concentration of ECs in the river water in the dry season was about 1.5 times higher than that in the wet season. The concentrations of these ECs close to the contaminated tributary and the WWTP were relatively high. Risk assessment showed that the maximum risk quotient value of estrone was 1.07 in the river water and estriol was 2.10 in the effluents. In addition, erythromycin posed generally medium risk in the river water and WWTP effluents. It should be paid attention to the prior control of the three contaminants in the river region.

In this study, we investigate the effect of application rate and timing of liquid swine slurry on leaching of antibiotic-resistant bacteria (ARB) and their antibiotic resistance genes (ARGs) through soil columns. Swine slurry was added to laboratory soil columns at rates of 5000 or 30,000 gal ac⁻¹. For both application rates, rainfall was applied at either 1, 7, or 21 days after slurry application. Column effluent and the top centimeter of soil in the columns were sampled post-rainfall for cultivable bacteria and quantitative PCR was used to quantify tetracycline, methicillin, β-lactam, and erythromycin resistance genetic determinants. We also conducted similar experiments using swine lagoon slurry spiked with antibiotic-resistant E. coli and Salmonella. We found that the concentration of ARB and ARG recovered in the column effluent following application of the swine lagoon slurry generally decreased with increasing lag time between slurry application and simulated rainfall, though most of these decreases were not statistically significant. Moreover, no statistically significant differences in CFU or GU concentration in the column leachate were observed between the low and high slurry application rates. In the experiments using swine slurry spiked with E. coli and Salmonella, concentrations of both microorganisms eluted from fine sand columns were affected by both slurry application rate and lag time; recovery of ARGs, however, was mostly unaffected, but some differences were observed. In columns packed with loamy sand, no recovery was detected in the column effluent for either organism and recovery of ARG was unaffected by manure application rate or rainfall lag time.

Marine fish species were analyzed for culturable and total metagenomic microbial diversity, antibiotic resistance (AR) pattern, and horizontal gene transfer in culturable microorganisms. We observed a high AR microbial load of 3 to 4 log CFU g⁻¹. Many fish pathogens like Providencia, Staphylococcus, Klebsiella pneumoniae, Enterobacter, Vagococcus, and Aeromonas veronii were isolated. Photobacterium and Vibrio were two major fish and human pathogens which were identified in the fish metagenome. Other pathogens that were identified were Shewanella, Acinetobacter, Psychrobacter, and Flavobacterium. Most of these pathogens were resistant to multiple antibiotics such as erythromycin, kanamycin, neomycin, streptomycin, penicillin, cefotaxime, bacitracin, rifampicin, trimethoprim, ciprofloxacin, and doxycycline with a high multiple antibiotic resistance index of 0.54–0.77. The fish microflora showed high prevalence of AR genes like bla TEM, Class I integron, tetA, aph(3′)-IIIa, ermB, aadA, and sul1. Nineteen of 26 AR isolates harbored Class I integrons showing high co-resistance to trimethoprim, kanamycin, doxycycline, and cefotaxime. Mobile R-plasmids from 6 of the 12 AR pathogens were transferred to recipient E. coli after conjugation. The transconjugants harbored the same R-plasmid carrying bla CTX₋M, dfr1, tetA, bla TEM, and cat genes. This study confirms that fish is a potential carrier of AR pathogens which can enter the human gut via food chain. To the best of our knowledge, this is the first study in the Indian subcontinent reporting a direct evidence of spread of AR pathogens to humans from specific marine fish consumption.