Effects of Single and Combined Ciprofloxacin and Lead Treatments on Zebrafish Behavior, Oxidative Stress, and Elements Content.

Even though the toxic effects of antibiotics and heavy metals have been extensively studied in the last decades, their combined adverse impact on aquatic organisms is poorly understood. Therefore, the objective of this study was to assess the acute effects of a ciprofloxacin (Cipro) and lead (Pb) mixture on the 3D swimming behavior, acetylcholinesterase (AChE) activity, lipid peroxidation level (MDA-malondialdehyde), activity of some oxidative stress markers (SOD-superoxide dismutase and GPx-glutathione peroxidase), and the essential elements content (Cu-copper, Zn-zinc, Fe-iron, Ca-calcium, Mg-magnesium, Na-sodium and K-potassium) in the body of zebrafish (Danio rerio). For this purpose, zebrafish were exposed to environmentally relevant concentrations of Cipro, Pb, and a mixture for 96 h. The results revealed that acute exposure to Pb alone and in mixture with Cipro impaired zebrafish exploratory behavior by decreasing swimming activity and elevating freezing duration. Moreover, significant deficiencies of Ca, K, Mg, and Na contents, as well as an excess of Zn level, were observed in fish tissues after exposure to the binary mixture. Likewise, the combined treatment with Pb and Cipro inhibited the activity of AChE and increased the GPx activity and MDA level. The mixture produced more damage in all studied endpoints, while Cipro had no significant effect. The findings highlight that the simultaneous presence of antibiotics and heavy metals in the environment can pose a threat to the health of living organisms.

Microplastics exacerbate co-occurrence and horizontal transfer of antibiotic resistance genes.

Microplastic pollution is a rising environmental issue worldwide. Microplastics can provide a niche for the microbiome, especially for antibiotic-resistant bacteria, which could increase the transmission of antibiotic resistance genes (ARGs). However, the interactions between microplastics and ARGs are still indistinct in environmental settings. Microplastics were found to be significantly correlated with ARGs (p < 0.001), based on the analysis of samples taken from a chicken farm and its surrounding farmlands. Analysis of chicken feces revealed the highest abundance of microplastics (14.9 items/g) and ARGs (6.24 ×108 copies/g), suggesting that chicken farms could be the hotspot for the co-spread of microplastics and ARGs. Conjugative transfer experiments were performed to investigate the effects of microplastic exposure for different concentrations and sizes on the horizontal gene transfer (HGT) of ARGs between bacteria. Results showed that the microplastics significantly enhanced the bacterial conjugative transfer frequency by 1.4-1.7 folds indicating that microplastics could aggravate ARG dissemination in the environment. Potential mechanisms related to the up-regulation of rpoS, ompA, ompC, ompF, trbBp, traF, trfAp, traJ, and down-regulation of korA, korB, and trbA were induced by microplastics. These findings highlighted the co-occurrence of microplastics and ARGs in the agricultural environment and the exacerbation of ARGs' prevalence via rising the HGT derived from microplastics.

A new insight into the potential drivers of antibiotic resistance gene enrichment in the collembolan gut association with antibiotic and non-antibiotic agents.

Effects of non-antibiotic pharmaceuticals on antibiotic resistance genes (ARGs) in soil ecosystem are still unclear. In this study, we explored the microbial community and ARGs variations in the gut of the model soil collembolan Folsomia candida following soil antiepileptic drug carbamazepine (CBZ) contamination, while comparing with antibiotic erythromycin (ETM) exposure. Results showed that, CBZ and ETM all significantly influenced ARGs diversity and composition in the soil and collembolan gut, increasing the relative abundance of ARGs. However, unlike ETM, which influences ARGs via bacterial communities, exposure to CBZ may have primarily facilitated enrichment of ARGs in gut through mobile genetic elements (MGEs). Although soil CBZ contamination did not pose an effect on the gut fungal community of collembolans, it increased the relative abundance of animal fungal pathogens contained therein. Soil ETM and CBZ exposure both significantly increased the relative abundance of Gammaproteobacteria in the collembolan gut, which may be used to indicate soil contamination. Together, our results provide a fresh perspective for the potential drivers of non-antibiotic drugs on ARG changes based on the actual soil environment, revealing the potential ecological risk of CBZ on soil ecosystems involving ARGs dissemination and pathogens enrichment.

Microbial profiles and immune responses in seahorse gut and brood pouch under chronic exposure to environmental antibiotics.

Ocean antibiotics pose substantial risks to the adaptation and lifespan of marine organisms. Seahorses are unique owing to the occurrence of brood pouches, male pregnancy, and loss of gut-associated lymphatic tissues and spleen, which lead to increased sensitivity to environmental changes. This study evaluated the changes in microbial diversity and immune responses within the gut and brood pouch in the lined seahorse Hippocampus erectus under chronic exposure to environmental levels of triclosan (TCS) and sulfamethoxazole (SMX), which are common antibiotics in coastal regions. The results showed that microbial abundance and diversity within the gut and brood pouch of seahorses were significantly changed following antibiotics treatment, with the expression of core genes involved in immunity, metabolism, and circadian rhythm processes evidently regulated. Notably, the abundance of potential pathogens in brood pouches was considerably increased upon treatment with SMX. Transcriptome analysis revealed that the expression of toll-like receptors, c-type lectins, and inflammatory cytokine genes in brood pouches was significantly upregulated. Notably, some essential genes related to male pregnancy significantly varied after antibiotic treatment, implying potential effects on seahorse reproduction. This study provides insights into the physiological adaptation of marine animals to environmental changes resulting from human activity.

Study effect and mechanism of levofloxacin on the neurotoxicity of Rana nigromaculata tadpoles exposed to imidacloprid based on the microbe-gut-brain axis.

Aquatic organisms may be simultaneously exposed to antibiotics and pesticides. After levofloxacin (LVFX), imidacloprid (IMI) exposure and co-exposure at environmental levels, we found LVFX and IMI had antagonistic effect on the neurotoxicity of tadpoles. IMI-induced neurotoxicity on tadpoles can be explained by oxidative stress and hormone levels in some degree. By regulating ornithine, l-asparagine, putrescine and tryptamine in the intestine, LVFX affected glutathione metabolism, arginine and proline metabolism, alanine, aspartate and glutamate metabolism, tyrosine metabolism and aminoacyl tRNA biosynthesis, so then eased the neurotoxicity caused by IMI. More interestingly, Fusobacteriota and Cetobacterium might play an important role on easing the neurotoxicity caused by IMI. In addition, LVFX might have a laxation effect on the increased relative abundance of Bacteroidota caused by IMI. In conclusion, IMI not only affected oxidative stress and hormone levels in the brain, but also affected the synthesis of neurotransmitters in the intestine by regulating intestinal microbiota. In LVFX and IMI co-exposed groups, LVFX alleviated the neurotoxicity caused by IMI through regulating the intestinal microbiota, showing as an antagonistic effect. Our results provided a new perspective for aquatic ecological risk assessment under co-exposure of antibiotics and pesticides.

Synthesis, applications, toxicity and toxicity mechanisms of silver nanoparticles: A review.

Silver nanoparticles (AgNPs) have become one of the most popular objects of study for the past few decades. The ability to design AgNPs through different synthetic methods according to the application area and desired features is their advantage in many applications. Green synthesis of silver nanoparticles has become one of the most potential synthesis methods. Because of their strong antibacterial activity, AgNPs have been used in a wide range of applications, such as food packaging and medical products and devices. With the increasing application of AgNPs, it is becoming necessary for a better understanding of the toxicity of AgNPs and their potential mechanism of toxicity. In the review, we first describe the synthetic methods of AgNPs. The application of AgNPs in the field is then briefly described. The toxicity of AgNPs and their potential toxicity mechanisms are discussed.

Chitosan Sponge/Cu-WO3-x Composite for Photodynamic Therapy of Wound Infection.

One of the potential treatments for bacterial wound infections is photodynamic therapy. WO3-x semiconductor materials can generate reactive oxygen species when exposed to light, which can inactivate bacteria. In this work, we improved their photocatalytic performance by doping WO3-x with Cu. The wound dressing was prepared by loading Cu-WO3-x into a highly biocompatible chitosan sponge. The composite sponge dressing showed significant inactivation of Escherichia coli and Staphylococcus aureus, and in vitro toxicity assays on L929 cells demonstrated the biosafety of the dressing. Through in vivo wound healing trials, composite sponge dressings have been shown to accelerate wound healing, and this composite chitosan sponge can be possibly used for photodynamic therapy of bacterial wound infections.

Response of submerged macrophytes and biofilms to coexisting azithromycin and tetracycline: Antibiotic resistance genes removal, toxicity assessment and microbial properties.

Antibiotics, such as azithromycin (AZ), tetracycline (TC), and their related antibiotic resistance genes (ARGs), create serious ecological risks to aquatic organisms. This study examined the response mechanisms of submerged macrophytes and periphytic biofilms to a mixture of AZ and TC pollution and determined the antibiotic removal efficiencies and fate of ARGs. The results showed that the plant-biofilm system had a significant capacity for removing both single and combined antibiotics with removal efficiencies of 93.06% ∼99.80% for AZ and 73.35% ∼97.74% for TC. Higher ARG (tetA, tetC, tetW, ermF, ermX, and ermB) abundances were observed in the biofilm, and subsequent exposure to the antibiotic mixture increased the abundances of these genes. Both single and combined antibiotics triggered antioxidant stress, but antagonistic effects were induced only with mixed AZ and TC exposure. Furthermore, the antibiotics changed the structural characteristics of extracellular polysaccharides and induced alterations in the structure of the biofilm microbial community. Increased N-acylated-l-homoserine lactone confirmed alternations in microbial quorum-sensing. The results extend the understanding of the fate of antibiotics and ARGs when aquatic plants and biofilms are exposed to antibiotic mixtures, as well as the organism's response mechanisms.

Mechanisms for the increase in lipid production in cyanobacteria during the degradation of antibiotics.

This study evaluated the responses of cell density, photosynthesis activity, dry cell weight, lipid productivity, proteome and metabolome in two non-toxic cyanobacterial species (Synechococcus sp. and Chroococcus sp.) exposed to two frequently detected antibiotics (sulfamethoxazole and ofloxacin) at test concentrations of 0.2-20.0 µg L-1 in a 4-day culture period. Upregulated antioxidant enzymes and oxidoreductases contributed to antibiotic biodegradation in Synechococcus sp.; whereas, upregulated carotenoid protein contributed to antibiotic biodegradation in Chroococcus sp. The 4-day removal efficiencies of sulfamethoxazole and ofloxacin by cyanobacteria were 35.98-66.23% and 33.01-61.92%, respectively. In cyanobacteria, each antibiotic induced hormetic responses, such as increase in cell density, dry cell weight, and photosynthetic activity; upregulation of photosynthesis-related proteins; and elevation of lipid expression by up to 2.05-fold. Under antibiotic stress, the two cyanobacterial species preferred to store energy in the form of lipids rather than ATP, with fructose-bisphosphate aldolase playing an essential role in lipid synthesis. The downregulation of lipid transporters also facilitated lipid accumulation in Synechococcus sp. In general, the two non-toxic cyanobacterial species achieved a good combination of lipid deposition and antibiotic treatment performance, especially in Chroococcus sp. exposed to sulfamethoxazole.

Comparative metagenomics reveals poultry and swine farming are hotspots for multidrug and tetracycline resistance.

Antibiotic misuse in livestock is a major threat to human health, as bacteria are quickly developing resistance to them. We performed a comparative analysis of 25 faecal metagenomes from swine, poultry, cattle, and humans to investigate their resistance profiles. Our analysis revealed that all genes conferring resistance to antibiotic classes assessed except tetracyclines were more prevalent in poultry manure than in the remaining species. We detected clinically relevant antibiotic resistance genes, such as mcr-1 which confers resistance to polymyxins. Among them, extended-spectrum ß-lactamase blaCTX-M genes were particularly abundant in all species. Poultry manure was identified as a hotspot for multidrug resistance, which may compromise medical treatment options. Urgent actions in the livestock industry are imperative to hamper the emergence and spread of antibiotic resistance.

The role and mechanism of quorum sensing on environmental antimicrobial resistance.

As more environmental contaminants emerging, antibiotics and antibiotic resistance genes (ARGs) have caused a substantial increase of antimicrobial resistance (AMR) in environment. Quorum sensing (QS) is a bacterial cell-to-cell communication process that regulates many traits and gene expression, including ARGs and the related genes that contribute to AMR development. Herein, we summarize the role, physiology, and genetic mechanisms of bacterial QS in AMR development in the environment. First, the effect of QS on AMR is introduced. Next, the role of QS in bacterial physiological behaviors that promote AMR development, including membrane permeability, tactic movement, biofilm formation, persister formation, and small colony variants (SCVs), is systematically analyzed. Furthermore, the regulation of QS on the expression of ARGs, generation of reactive oxygen species (ROS), which affects ARGs formation, and horizontal gene transfer (HGT), which accelerates the transmission of ARGs, are discussed to reveal the molecular mechanism for AMR development. This review provides a reference for a better understanding of AMR evolution and novel insights into AMR prevention.

Immobilization, oxidative stress and antioxidant response of Daphnia magna to Amoxicillin and Ciprofloxacin.

Amoxicillin (AMX) and ciprofloxacin (CPX) are broad-spectrum antibiotics with wide application in agriculture and human and veterinary medicine. The drugs end up in the environment where their impact on zooplankton remains scantily understood. This study investigated the immobilization, risk assessment (RQ), antioxidant response, and biochemical changes of Daphnia magna post-exposure to AMX and CPX. Sixty-percent immobilization of Daphnia occurred at 200 µg L-1 AMX and CPX, while EC50 values were 2391.6 µg L-1 and 273.4 µg L-1, respectively. RQs were 113.3 and 11,481.5, while Toxic units were 41.6 and 364.9 for AMX and CPX, respectively. Both antibiotics caused a significant rise in intracellular hydrogen peroxide 48 h post-exposure, indicating oxidative stress. Lipid peroxidation and antioxidant enzyme activity were considerably altered during the research. Thus, environmentally relevant concentrations of AMX and CPX pose an adverse risk that could change the population dynamics of Daphnia magna.

Optimizing the multimedia fate model for characterizing environmental risks of florfenicol in seasonally ice-covered reservoirs.

Seasonally ice-covered reservoirs have both freeze-thaw and artificial regulation characteristics which could cause the accumulation of antibiotics. Florfenicol, one of the most widely used veterinary antibiotics, with an environmental persistence due to its fluorinated substituents has been detected in the suburban drinking water source reservoirs. In this study, a four-level fugacity model that is appropriate for ice-water-sediment systems was developed to predict the fate of florfenicol and assess its ecological risk in seasonally ice-covered reservoirs. The effects of freeze-thaw and artificial regulation processes on the volume variation of ice and water were considered by the model. The simulation accuracies in ice and water in the model were improved by 3.9% and 17.7%, respectively, compared with the traditional model. The results of mass transfer analysis showed that the inflow of florfenicol in tributaries and the volume variation of ice and water were the major factors influencing the concentration variation of florfenicol in the seasonally ice-covered reservoir. Additionally, ecological risk analysis showed that the values of risk quotients ranged from 0.019 to 0.038 which was consistently at a low ecological risk level. Our findings provide a modeling tool for predicting the fate of antibiotics with persistence and assessing their ecological risks in seasonally freeze-thaw reservoirs in cold regions.

Cationic Gelatin Cross-Linked with Transglutaminase and Its Electrospinning in Aqueous Solution.

Gelatin (GE) is a renewable biopolymer with abundant active groups that are beneficial for manufacturing functional biomaterials via GE modification. An antibacterial fibrous GE film was prepared by electrospinning the modified GE in an aqueous solution. The original GE was modified by reacting it with N,N-dimethyl epoxypropyl octadecyl ammonium chloride (QAS), and then it was cross-linked with transglutaminase (TGase). FTIR analysis illustrated that QAS was grafted onto GE through the epoxy ring-opening reaction, and the modification did not influence the main GE skeleton structure. The investigation of the solution properties showed that the grafted cationic QAS group was the main factor that decreased the surface tension of the solution, increased the electrical conductivity of the solution, and endowed GE with antibacterial activity. TGase cross-linking clearly influenced the rheological properties such that the flow pattern of the spinning solution varied from Newton-type to shear thinning, and the aqueous solution of GE-QAS-TGs transformed from liquid-like to solid-like and even induced gelatinization with increasing TGase content. A satisfactory fibrous morphology of 200-500 nm diameter was obtained using a homemade instrument under the optimized electrospinning conditions of a temperature of 35 °C, a distance between electrodes of 12 cm, and a voltage of 15 kV. The study of film properties showed that the antibacterial activity of the fibrous GE film depended only on the grafted quaternary ammonium, whereas the thermostability, wettability, and permeability were greatly influenced by both the TGase cross-linking and film-forming methods. Cytotoxicity was tested using the CCK-8 and live/dead kit staining methods in vitro, which showed that the modified GE had good biocompatibility.

Predation risk affects the ecotoxicity evaluation of antibiotics: Population growth and antioxidase activity in the ciliate Paramecium jenningsi.

Although predation risk exists under natural conditions, its role is usually ignored when evaluating the ecotoxicity of environmental contaminants, and the interaction between predation risk and antibiotic ecotoxicity is not yet clear. To investigate the nonconsumptive effects (NCEs) of predation on the ecotoxicity evaluation of antibiotics, the median lethal concentration (LC50), relative population growth rate (RGR), and activities of three antioxidases were measured in the ciliate Paramecium jenningsi exposed to graded concentrations of the antibiotics nitrofurazone (NFZ) or erythromycin (ERY) in the presence or absence of a predator, i.e., the ciliate Didinium nasutum. The results showed that (1) NCEs significantly reduced the LC50 of NFZ but had no effect on that of ERY; (2) predation pressure alone had no significant effect on the inhibitory rate of the P. jenningsi population, but the interaction with NFZ was synergistic, while that with CRY was additive; (3) the concentrationresponse (i.e., mortality) model for each antibiotic exposure with and without predation pressure differed significantly in the parameter slope; (4) RGRs were significantly reduced by antibiotic exposure or NCEs; only in NFZ-exposed groups did the RGRs decrease linearly with increasing exposure concentration; and (5) the activities of all three antioxidases significantly increased due to NCEs or following exposure to antibiotics. In brief, NCEs were detected in P. jenningsi, and these had additive or synergistic effects on antibiotic ecotoxicity, but their magnitude depended on the properties and exposure concentrations of the antibiotics. Our findings suggest that it is necessary to consider the roles of NCEs in the ecotoxicity evaluation of environmental contaminants.

Photocatalysis Combined with Microalgae to Promote the Degradation and Detoxification of Tetracycline Hydrochloride.

The continuous discharge of antibiotics into the environment poses a serious threat to the ecological environment and human health. In this study, photocatalysis and microalgae were combined to study the removal of tetracycline hydrochloride (TCH) and its photodegradation intermediates in water. The results showed that after photocatalytic treatment, the removal rate of TCH reached 80%, but the mineralization rate was only 17.7%. While Chlorella sp. alone had poor tolerance to high concentrations of TCH, the combined treatment of photocatalysis and microalgae completely removed TCH and increased the mineralization efficiency to 35.0%. Increased cell density was observed, indicating that TCH and the intermediates produced in the photocatalysis process could be utilized by algae for growth. Meanwhile, TCH degradation pathways were proposed based on Liquid Chromatograph Mass Spectrometer analysis, and the toxicity of intermediates detected was predicted using ECOSAR software, which showed that the type and quantity of highly toxic intermediates decreased significantly after subsequent algal treatment. The results demonstrate that photocatalysis and microalgae combined treatment is an efficient and eco-friendly method for the removal of antibiotics in water.

The different toxicological effects and removal efficiencies of norfloxacin and sulfadiazine in culturing Arthrospira (Spirulina) platensis.

Norfloxacin (NFX) and sulfadiazine (SDZ) are two widely used antibiotics belonging to fluoroquinolone and sulfonamide groups, respectively, and have become the commonly detected micropollutants in aquatic environments. However, only few works have been conducted to investigate the highly probable inhibition of these antibiotic pollutants to Arthrospira platensis, which is an important species of cyanobacteria that is one of primary producers in aquatic ecosystems and should be remarkably sensitive to environmental pollutants due to its prokaryotic characteristics. Hence, the toxicological effects and removal efficiencies of NFX and SDZ in culturing A. platensis were studied by analyzing the biomass growth, photosynthetic pigments, primary biocomponents, and antibiotics concentration. The corresponding variations of these characteristics showed the higher sensitivity of A. platensis to NFX than to SDZ, indicating the specifically targeted effect of NFX on A. platensis, which could be confirmed in silico by the higher binding affinity of NFX with the critical enzyme. The obtained results illustrated the roles of NFX and SDZ on the growth of A. platensis, thus providing the great support in employing A. platensis to reduce hazards from contaminated water and recover biomass resources.

Antibiotics modulate frequency and early generation of epileptic seizures in zebrafish.

Antibiotics have been used for decades to treat various bacterial infections. Apart from bactericidal activities, their potential side effects have not been much studied or evaluated. Neurotoxicity is a major concern in the case of ß-lactam and fluoroquinolone families, which can result in convulsions or seizures. Here, we proposed a hypothesis to check whether antibiotic treatment can conclusively enhance anxiety-like behaviours and how seizure behavioural profile gets modulated in pentylenetetrazole (PTZ)-treated zebrafish. Zebrafish were treated with selected antibiotics such as 25 mg/L Penicillin G (PG) and Ciprofloxacin (CPFX), for 7 days and thereafter exposed to PTZ (7.5 mM) for 20 min. The data indicate that PG and CPFX-treated groups exhibited anxiety-like or stressed behavioural phenotypes in the novel tank test (6 min), and also, they were found to promote hyperactivity. Early onset of PTZ-induced seizure-like behavioural scores, the heightened intensity of seizure and reduced latency in different scores were found in PG and CPFX-administered groups. This study substantiates that PG and CPFX as potential seizure modulators in zebrafish. The zebrafish is a well-established and still expanding model organism in many fields. Here, we again reinforce zebrafish as a prominent model to investigate seizure-like neuro-behavioural entities and confirm that chronic antibiotic use has negative consequences that can exacerbate the circumstances of vertebrate species exhibiting seizure-related reactions.

Evaluating the impact of heavy metals on antimicrobial resistance in the primary food production environment: A scoping review.

Heavy metals are naturally occurring environmental compounds, which can influence antimicrobial resistance (AMR) dissemination. However, there is limited information on how heavy metals may act as a selective pressure on AMR in the primary food production environment. This review aims to examine the literature on this topic in order to identify knowledge gaps. A total of 73 studies, which met pre-established criteria, were included. These investigations were undertaken between 2008 and 2021, with a significant increase in the last three years. The majority of studies included were undertaken in China. Soil, water and manure were the most common samples analysed, and the sampling locations varied from areas with a natural presence of heavy metals, areas intentionally amended with heavy metals or manure, to areas close to industrial activity or mines. Fifty-four per cent of the investigations focused on the analysis of four or more heavy metals, and copper and zinc were the metals most frequently analysed (n = 59, n = 49, respectively). The findings of this review highlight a link between heavy metals and AMR in the primary food production environment. Heavy metals impacted the abundance and dissemination of mobile genetic elements (MGEs) and antimicrobial resistance genes (ARGs), with MGEs also observed as playing a key role in the spread of ARGs and metal resistance genes (MRGs). Harmonization of methodologies used in future studies would increase the opportunity for comparison between studies. Further research is also required to broaden the availability of data at a global level.

Assessment of toxic mechanisms and mode of action to three different levels of species for 14 antibiotics based on interspecies correlation, excess toxicity, and QSAR.

Antibiotics have received much attention owing to their ecotoxicity toward nontarget aquatic creatures. However, the mode of action (MOA) of toxicity against nontarget organisms is unclear in some aquatic organisms. In this study, the comparison of toxicities through interspecies correlations, excess toxicity calculated from toxicity ratio, and quantitative structure-activity relationship (QSAR) was carried out to investigate the MOAs for 14 antibiotics among Daphnia magna, Vibrio fischeri, and Pseudokirchneriella subcapitata. The results showed that interspecies toxicity correlations were very poor between any two of the three species for the 14 antibiotics. The toxicity ratio revealed that most antibiotics exhibited excess toxicity to algae and Daphnia magna but not to V. fischeri, demonstrating that some antibiotics share the same MOA, but some antibiotics share different MOAs among the three different levels of species. P. subcapitata was the most sensitive species, and V. fischeri was the least sensitive species. This is because of the differences in the biouptake and interactions of antibiotics with the target receptors between the three different trophic levels of the species. Molecular docking simulations suggested that the toxicity of antibiotics depends highly on their interactions with target receptors through hydrogen bonds, electrostatic or polar interactions, π bond interactions, and van der Waals forces. QSAR models demonstrated that hydrogen bonding and electrophilicity/nucleophilicity play key roles in the interaction of antibiotics with different receptors in the three species. The toxic mechanisms of antibiotics are attributed to the interactions between electrophilic antibiotics and biological nucleophiles, and hydrogen-bond interactions. These results are valuable for understanding the toxic mechanisms and MOA of the three different levels of species.