Search Results (333)

Kindergarten environments combine high microbial exposure with increased immunological vulnerability, yet antibiotic resistance genes (ARGs) in kindergarten dust remain poorly characterized. This systematic review synthesized evidence on the occurrence and potential health relevance of ARGs in kindergarten dust. Following PRISMA 2020 guidelines, PubMed, Scopus, and Web of Science were searched. Four studies from China, Hong Kong, and Norway (2018–2024) met the inclusion criteria. ARGs were detected in all kindergarten dust samples, indicating that dust is a consistent reservoir of antibiotic resistance determinants. A consensus resistome (classes detected in ≥2 studies) encompassed sulfonamide, macrolide–lincosamide–streptogramin B (MLSB), tetracycline, beta-lactam, aminoglycoside, and multidrug resistance genes; beta-lactam resistance genes were the only class reported in all four studies. Clinically important ARGs associated with last-resort antibiotics, including mecA, vanA, bla_NDM, and mcr-5, were reported in three studies. Class 1 integron-integrase genes (intI1) frequently co-occurred with ARGs, suggesting potential horizontal gene transfer. Limited evidence indicated higher ARG abundance in urban and winter samples. One study reported antibiotic-resistant bacteria carrying resistance markers concordant with those in kindergarten dust in the urine of children attending the same facilities; however, this cross-sectional, single-site evidence is consistent with, but not sufficient to establish, a dust-to-child exposure pathway. The available evidence supports the plausibility that kindergarten dust may contribute to children’s exposure to ARGs and ARG-carrying bacteria, but current studies do not establish causal transmission from dust to child colonization or infection. Standardized monitoring and longitudinal studies are needed to assess health risks and guide mitigation strategies in early childhood educational settings. Full article

Sulfonamide antibiotics threaten ecosystems, yet their phytotoxic mechanisms remain elusive. Here, we demonstrate that sulfonamides inhibit root growth by disrupting the ROS-ethylene signaling axis. Sulfadiazine (SD) strongly inhibits root elongation, an effect strictly dependent on ACS1, as the acs1-1 mutant is completely insensitive. Mechanistically, SD triggers an RBOH-dependent ROS burst that transcriptionally induces ACS1. Since ACS1 forms active heterodimers with ACS2/ACS6, acs2-1 and acs6-1 mutants exhibit partial insensitivity. Furthermore, SD partially depends on the MPK3/6 cascade to post-translationally stabilize ACS2/ACS6, with mpk3 and mpk6 mutants showing partial resistance. Collectively, sulfonamides employ a dual-pronged mechanism: ROS transcriptionally activate ACS1, while MPK3/6 post-translationally stabilizes ACS2/ACS6, driving aberrant ethylene precursor ACC biosynthesis and root growth inhibition. Full article

Multidrug-resistant bacterial enteric pathogens such as Salmonella and Shigella have the potential to cause significant mortality and represent a major issue facing the global health community. This is particularly concerning in low-and middle-income countries where access to clean water and antimicrobials is limited. We aimed to determine the frequency, antimicrobial resistance, and presence of resistance genes among Salmonella, Shigella, Vibrioand Campylobacter recovered from patients presenting with diarrhea in an informal settlement in Nairobi, Kenya, from 2018 to 2020. Conventional bacteriologic methods were used for bacterial culture and isolation. BD Phoenix M50 technology was used for the identification and speciation of bacteria recovered from stool samples and for determining minimum inhibitory concentrations to both clinically relevant antimicrobials and agents of epidemiologic significance. PCR testing was subsequently performed to identify resistance genes. The key pathogenic bacteria recovered were: Shigella flexneri (58.3%), Salmonella enterica serovar Typhi (8.3%), Shigella dysenteriae (6.7%), Shigella boydii (6.7%), and Shigella sonnei (6.7%); no Campylobacter or Vibrio were isolated. Overall resistance among Shigella spp. and Salmonella spp. was determined to be highest to ampicillin (83%), followed by tetracycline (50%), cotrimoxazole (47%), chloramphenicol (17%), ceftriaxone (7%), gentamycin (5%), meropenem (2%), ciprofloxacin (0%) and amoxicillin-clavulanic acid (2.0%). Resistance genes detected included β-lactamases (bla_TEM and bla_SHV), aminoglycoside 3-N acetyltransferase acc(3), sulfonamide resistance genes (sul1, sul2), and tetracycline resistance genes tet(A) and tet(B). Results emphasize the critical need for active and continuous surveillance of pathogenic enteric bacteria to improve patient management and inform the development of empiric therapy guidelines. Full article

Background/Objectives: Microtubule-targeting agents represent a pillar of cancer chemotherapy; however, their clinical utility is constrained by significant toxicity, pharmacokinetic instability, and susceptibility to multidrug resistance transporters. This study aimed to explore the impact of replacing substituted phenyl rings with a naphthalene moiety in sulfonamide-based colchicine-site ligands, with the goal of identifying new antiproliferative candidates with improved profiles. Methods: We designed, synthesized, and evaluated a library of 35 naphthalene sulfonamides bearing varied aryl groups and sulfonamide nitrogen substituents. We assessed the antiproliferative activity against multiple cancer cell lines. Mechanistic studies, including fluorescence microscopy, cell cycle analysis, and cell death assays, were performed to evaluate the effect of these compounds on microtubule polymerization dynamics and cell fate. Molecular docking and in silico pharmacokinetic profiling were carried out to support the proposed binding mode at the colchicine site and to assess drug-likeness. Results: Exclusively, compounds bearing a trimethoxyphenyl group showed antiproliferative activity in the submicromolar range, thus identifying it as a structural requirement. The most potent compound (2) reached double-digit nanomolar IC₅₀ values (67–104 nM) across multiple cancer cell lines. Microscopy confirmed intracellular disruption of microtubule polymerization. Unlike colchicine, these compounds did not induce canonical mitotic arrest but instead triggered apoptotic cell death. In silico analyses supported binding at the colchicine site and revealed favorable predicted pharmacokinetic properties. Conclusions: The naphthalene sulfonamides described herein demonstrate potent antiproliferative activity through a distinct mechanism compared to colchicine, and their favorable in silico profiles position them as promising candidates for further development as antitumor agents. Full article

The widespread use of antimicrobials in poultry production exerts selective pressure for the emergence of resistant bacteria, yet molecular data on resistance determinants in Escherichia coli from commercial farms in Central Asia remain scarce. This study aimed to characterize antimicrobial resistance genes and mobile genetic elements in E. coli isolates from a layer poultry farm in the West Kazakhstan region. A total of 100 samples (cloacal swabs, fresh feces, internal organs, and wild pigeons) were collected. E. coli was isolated using selective media and confirmed by qPCR targeting the uidA gene. Forty isolates were confirmed, of which ten (25%) carried at least one resistance gene as determined by a panel of 57 primer sets. All ten isolates carrying antimicrobial resistance-associated genes (100%) harbored determinants associated with multidrug resistance. The most frequently detected genes were blaTEM (80%), aadA17 (90%), and intI1-a-marko (80%). A high prevalence of qacEΔ1 (80%) was also observed. At least one class 1 integron-associated gene was detected in all resistant isolates and those carrying both integron variants harbored the highest numbers of resistance genes (14–19). Resistance gene profiles in pigeon isolates mirrored those from poultry, suggesting wild birds as potential vectors. The correlation between detected resistance genes and farm antimicrobial use (enrofloxacin, trimethoprim/sulfonamide combinations) indicates strong selective pressure. These findings suggest that poultry farms may serve as environments supporting the persistence and dissemination of antimicrobial resistance-associated genes in E. coli and underscore the need for enhanced biosecurity and antimicrobial stewardship within a One Health framework. Full article

A silent pandemic is underway that is causing more than a million deaths each year worldwide: antimicrobial resistance. According to World Health Organization forecasts, this phenomenon could become the leading cause of death by 2050. It represents one of the most pressing global health threats, compromising the effectiveness of life-saving therapies and making infections increasingly difficult to treat. Although antimicrobial resistance is often considered a recent phenomenon, it was first recognized in the early stages of antimicrobial use, with reports predating Alexander Fleming’s discovery of penicillin in 1928. In this context, the present review examines selected representative cases of the early recognition of antimicrobial resistance through the analysis of the initial evidence of microbial adaptation associated with the use of pre-antibiotic chemotherapeutic agents such as Salvarsan, as well as the first reports of reduced clinical efficacy of sulfonamides and penicillin. These observations have contributed to the current understanding of resistance as a dynamic process driven by microbial adaptive mechanisms and amplified by the selective pressure exerted by antimicrobial use. Full article

Background/Objectives: Klebsiella (K.) pneumoniae is a significant pathogen in both humans and animals. However, data on its occurrence in animals in Germany remain limited. This study aimed to investigate the antimicrobial resistance (AMR) phenotypes, AMR genes, and virulence traits of animal-derived K. pneumoniae isolates from Germany. Methods: A total of 59 K. pneumoniae isolates obtained in 2023 from dogs, cats, horses, cattle, and chickens across 11 German federal states were analysed. Phenotypic antimicrobial susceptibility testing (AST) was performed, and whole-genome sequencing (WGS) was used for genomic characterisation, including detection of AMR genes, virulence-associated genes, sequence types (STs), and plasmid replicons. Results: Most isolates (78%) were susceptible to all tested antibiotics, while three isolates were classified as multidrug-resistant (MDR). Resistance was most frequently observed for piperacillin (n = 8) and trimethoprim/sulfamethoxazole (n = 4). Carbapenem resistance was detected in two isolates (one from a dog and one from a cat), and phenotypic colistin resistance in one dog isolate. WGS identified 96 AMR genes across isolates, with 20–42 AMR determinants per isolate, conferring resistance to β-lactams, aminoglycosides, fluoroquinolones, sulfonamides, tetracyclines, trimethoprim, and fosfomycin. Ten extended-spectrum β-lactamase (ESBL)-producing isolates carried genes including bla_CTX-M-15, bla_SHV-2, bla_SHV-27, bla_SHV-42, bla_SHV-106, and bla_TEM-158. Although fosA was detected in all isolates, only three exhibited phenotypic resistance to fosfomycin. A total of 52 STs were identified, including high-risk clones. One hypervirulent isolate (ST60) carrying hypervirulence-associated genes rmpA and iroB was detected. Plasmid replicons were present in 70% of isolates, while plasmid-associated AMR genes were identified in nine isolates. Conclusions: This study demonstrates the genomic diversity of K. pneumoniae identified in companion animals and highlights the presence of AMR and virulence determinants relevant to a One Health context. Full article

As a devastating disease worldwide, rice bacterial leaf blight—caused by Xanthomonas oryzae pv. oryzae (Xoo)—leads to substantial reductions in grain yield. The increasing resistance to conventional bactericides necessitates the development of novel and sustainable control agents. This study evaluated 58 novel alkyl sulfonamide compounds against Xoo. In the turbidimetric assay at 100 mg/L, several compounds showed potent antibacterial activity. Among them, SYAUP-116 and SYAUP-212 exhibited in vitro inhibition comparable to that of streptomycin sulfate at the same concentration. Furthermore, in EC₅₀ determination assays, both compounds yielded lower EC₅₀ values than zinc thiazole. Among the 58 compounds tested, SYAUP-491 exhibited an in vitro EC₅₀ of 6.96 mg/L and achieved 74.1% in vivo therapeutic efficacy at 200 mg/L, representing the most promising lead for further characterization. Molecular docking, based on prior proteomic data, indicates potential stable binding to ribosomal proteins (50S L33/L34 and 30S S5), with the strongest interaction observed for L33 (binding free energy: −5.73 kcal/mol). This suggests a putative mechanism involving ribosome targeting and protein synthesis inhibition, which may be facilitated by hydrophobic interactions and halogen bonds derived from its trifluoromethyl and sulfonamide groups. SYAUP-491 demonstrates significant potential as a novel bactericide for rice bacterial leaf blight, warranting further research on structure-activity optimization, target validation, and field performance. Full article

Background/objectives: Infections caused by multidrug-resistant (MDR) bacteria are becoming increasingly difficult to treat with recommended antimicrobials. Considering the critical and growing challenge of antimicrobial resistance (AMR), this study aims to evaluate the antimicrobial susceptibility patterns of Escherichia coli clinical isolates from dogs in Grenada. This research project consists of two distinct studies: a retrospective analysis of AMR in canine E. coli isolates collected between 2010 and 2020, and a cross-sectional study characterizing the genotypic AMR profiles of E. coli isolates obtained between April and June 2023. Methods: A retrospective analysis of antibacterial sensitivity test (ABST) reports from canine clinical samples submitted to the Small Animal Clinic at St. George’s University (SGU), St. George’s, Grenada, between 2010 and 2020 revealed a notable prevalence of AMR among canine E. coli isolates. To further investigate the underlying mechanisms of this resistance, the study analyzed canine E. coli isolates that exhibited phenotypic resistance in ABST assays. These isolates were subsequently screened for AMR-associated genes using polymerase chain reaction (PCR) and next-generation sequencing (NGS). Results: The retrospective study identified 153 canine clinical isolates positive for E. coli. The antimicrobial drugs, imipenem, cefotaxime and ciprofloxacin were found to be highly effective against these isolates. However, a gradual increase in AMR was observed for amoxicillin–clavulanic acid (34.88%), ampicillin–sulbactam (17.31%), cephalexin (43.08%), cefpodoxime (22.31%), cephalothin (68.42%), and doxycycline (37.04%). In the prospective study, PCR analysis of resistant canine E. coli isolates detected the tetA (577 bp) and blaTEM (686 bp) genes. These AMR determinants were further confirmed through analysis of NGS reads and assembled contigs. Additionally, NGS-based predictions identified genes associated with resistance to aminoglycosides and potentiated sulfonamides. Conclusions: This study demonstrates that E. coli from dogs in Grenada exhibits resistance to tetracycline and several β-lactam antimicrobials. These findings underscore the need for rational antimicrobial stewardship and continuous AMR surveillance in small animal practice within the region. Full article

Antimicrobial resistance (AMR) in Escherichia coli (E. coli) from food-producing animals constitutes a substantial public health concern. This study characterized antimicrobial resistance profiles, phylogenetic diversity, virulence-gene distribution, and plasmid-borne extended-spectrum β-lactamase (ESBL) determinants of E. coli isolates recovered from water buffaloes with subclinical mastitis. Among the 54 ESBL-producing E. coli isolates, all were resistant to ampicillin and cefotaxime. High resistance rates were also observed for cephalothin (75.9%), trimethoprim–sulfamethoxazole (74.0%), ceftiofur (70.4%), florfenicol (68.5%), and cefazolin (63.0%). Lower resistance was recorded for colistin sulfate (40.7%), enrofloxacin (33.3%), and gentamicin (25.9%). Phylogenetic analysis of ESBL producers identified phylogroup B1 (42.6%) as predominant, followed by groups A (29.6%) and D (25.9%). Multilocus sequence typing (MLST) revealed that ST50 (20.4%) was the most common sequence type, and serogroup O150 was dominant (70.4%). Virulence genes, such as iss (81.5%), astA (59.3%), and espP (38.9%), were frequently detected among ESBL isolates. ESBL genes were predominantly bla_CTX-M-1 (27.8%) in all isolates, while the narrow-spectrum β-lactamase genes bla_TEM-1 (55.6%) and bla_OXA-10 (14.8%) were also commonly co-detected. Bioinformatic analysis predicted that all ESBL genes were associated with plasmid-derived contigs, with the predicted plasmid size ranging from approximately 32 to 187 kb and belonging to IncFIB, IncFIA, IncI1, IncFIA + I1, and IncFII replicon types. Conjugation frequencies ranged from 4.8 × 10⁻⁷ to 4.1 × 10⁻², and plasmids were predicted to carry additional resistance genes mediating resistance to chloramphenicol (floR), sulfonamides (sul1, sul3), tetracyclines (tet(A) and tet(B)), and trimethoprim (dfrA1, dfrA12). The co-carriage of ESBL genes with additional antimicrobial resistance and virulence determinants suggests the potential role of water buffaloes as reservoirs of clinically relevant resistance traits that may disseminate through horizontal gene transfer. Full article

Background/Objectives: Acinetobacter baumannii is a critical opportunistic pathogen causing severe healthcare-associated infections, particularly in intensive care units. The global dissemination of carbapenem-resistant A. baumannii (CRAB) and its environmental persistence necessitate continuous genomic surveillance to monitor high-risk clones. Methods: We conducted whole-genome sequencing (WGS), core genome multi-locus sequence typing (cgMLST), and phylogenomic analyses on 26 CRAB isolates collected at the National Institute for Infectious Diseases (INMI) “Lazzaro Spallanzani” IRCCS (September 2023–September 2024). Antimicrobial resistance determinants, virulence-related genes, and capsular (KL) and lipooligosaccharide outer core (OCL) loci were characterized by interrogation of comprehensive bioinformatic pipelines. Results: All CRAB isolates displayed an extensively drug-resistant (XDR) phenotype, with a shared resistance pattern to carbapenems, aminoglycosides, fluoroquinolones, fosfomycin, and sulfonamides, while being susceptible only to colistin and cefiderocol. The carbapenemase gene bla_OXA-23 was detected in all CRAB isolates, together with clone-specific bla_OXA-51-like variants. For all isolates, the resistome profile fully matched the observed resistance phenotype. All isolates belonged to the International Clonal Lineage II (ICL II), Pasteur Sequence Type (ST) 2, and Oxford ST369, ST208, and ST455. Integration of cgMLST data with phylogenomic analyses and genome-based classification of KL and OCL loci revealed five distinct clusters, each one including nearly identical isolates, indicating both intra-hospital dissemination and possible inter-hospital transmission. Virulome profiling revealed heterogeneous repertoires of virulence-associated genes, resulting in cluster-specific patterns, while patristic analysis identified phylogenetic clusters linking the study isolates to other Italian and other European lineages. Conclusions: This study underscores the complex genomic landscape of CRAB in our setting, driven by the circulation of different ICL II clonal types, and reinforces the urgency of integrated genomic surveillance and robust antimicrobial stewardship to mitigate the spread of high-risk XDR A. baumannii clones. Full article

Background/Objectives: The Saker Falcon (Falco cherrug) is an endangered raptor species of ecological and conservation relevance. Despite its status, data regarding its microbiota and the prevalence of antimicrobial resistance (AMR) remain scarce, especially in Eastern Europe. This single-facility study aims to investigate the phenotypic and genotypic AMR profiles of Gram-negative bacteria isolated from captive Saker Falcons in Western Romania. Methods: Freshly voided fecal droppings were collected non-invasively from 40 clinically healthy Saker Falcons. Bacterial identification was performed using selective media and the VITEK^® 2 system. Antimicrobial susceptibility testing (AST) was conducted on a representative subset of 12 isolates. Selected resistance-associated genes were screened by conventional PCR. Results: Escherichia coli was the most prevalent 60% (n = 24/40), followed by Hafnia alvei 10% (n = 4/40) and Pseudomonas spp. 10% (n = 4/40). AST revealed phenotypic resistance among Enterobacteriaceae primarily to ampicillin 20% (n = 2/10), tetracycline 20% (n = 2/10), fluoroquinolones and sulfonamides 10% (n = 1/10), while susceptibility to imipenem 90% (n = 9/10) and gentamicin 90% (n = 9/10) remained high. The targeted resistance-associated genes were detected in selected phenotypically resistant isolates. PCR screening detected blaZ and ampC in 62.5% (n = 5/8) of tested isolates, blaOXA-61 in 37.5% (n = 3/8), blaOXA-51 in 25% (n = 2/8), tetK in 37.5% (n = 3/8), and gyrA in 12.5% (n = 1/8). The isolate used as the negative control, pansusceptible in AST, was confirmed negative for all targeted genes. Conclusions: This single-facility study provides baseline data on AMR traits in Gram-negative bacteria associated with Saker Falcons in Western Romania. Given the limited scale and isolate-based design of the study, the findings should be interpreted cautiously, but they support further investigation of wildlife-associated AMR within a One Health context. Full article

Background/Objectives: Multidrug-resistant (MDR) Escherichia coli has increasingly been recognized as a pathogen capable of causing severe systemic infections in various animal species. However, reports describing respiratory and septicemic infections caused by MDR E. coli in guinea pigs remain scarce. The objective of this report was to describe the clinical, pathological, and microbiological findings associated with a fatal infection in a domestic guinea pig. Case Study: A 10-month-old female guinea pig (Cavia porcellus), kept as a companion animal in a household environment, presented with acute respiratory distress, lethargy, and anorexia, progressing rapidly to death within approximately 36 h of onset. Post-mortem examination revealed severe pulmonary congestion, diffuse inflammatory lesions in the trachea, and generalized vascular congestion in multiple organs. Bacteriological cultures obtained from lung and bone marrow samples yielded pure growth of Escherichia coli. Identification was confirmed using MALDI-TOF mass spectrometry. Antimicrobial susceptibility testing demonstrated resistance to several antibiotic classes, including β-lactams, fluoroquinolones, tetracyclines, sulfonamides, and phenicols, while susceptibility was retained only to aminoglycosides. Molecular analysis revealed the presence of virulence genes involved in adhesion and iron acquisition, supporting the pathogenic potential of the isolate. Conclusions: This report highlights the ability of MDR E. coli to cause severe respiratory and systemic infections in guinea pigs. The findings underline the importance of early diagnosis, appropriate antimicrobial stewardship, and improved husbandry conditions in preventing such infections. From a One Health perspective, the circulation of resistant strains in companion animals may represent a potential risk for both environmental and human health. Full article

Background: Multidrug-resistant (MDR) Escherichia coli in intensive pig production represents a persistent animal health and One Health concern. Here, we integrated quantitative phenotypic susceptibility data with whole-genome sequencing (WGS) to characterize the resistome and its inferred genomic context (chromosomal vs. plasmid-predicted contigs and mobile genetic element (MGE)-proximal regions) in swine-associated MDR E. coli from Hungary. Methods: A total of 203 E. coli isolates from large-scale pig farms were tested by broth microdilution. Based on resistance-oriented screening from an extended-spectrum β-lactamase (ESBL)-screen-positive pool, 116 isolates were subjected to whole-genome sequencing (WGS) as a resistance-enriched subset. Resistance determinants were annotated using the Comprehensive Antibiotic Resistance Database (CARD). Results: Resistance-oriented screening indicated frequent β-lactamase activity and ESBL screening positivity (110/203 and 127/203 isolates, respectively), consistent with strong antimicrobial selection pressure in the source population. Across the full phenotypic panel, 78/203 isolates (38.4%) met the MDR definition (non-susceptible to ≥3 antimicrobial classes), with marked between-farm variation (p < 0.001) but no age-group effect (p = 0.75). Non-β-lactam minimum inhibitory concentration (MIC) distributions showed pronounced, site-dependent high-MIC “tails”, most notably for tetracyclines, trimethoprim–sulfamethoxazole, fluoroquinolones, and colistin. In the WGS cohort (n = 116), we detected 82 distinct resistance determinants (5433 total occurrences), featuring a conserved chromosomal backbone enriched for intrinsic multidrug resistance components and lipid A modification pathways, alongside common plasmid- and MGE-associated acquired ARG modules involving tetracycline (tetA/tetB), sulfonamide/trimethoprim (sul/dfrA), aminoglycoside-modifying enzymes, and phenicol determinants (floR/cat). High-priority mobile determinants were rare but present, including mcr-1 (3/116; plasmid-associated) and plasmid-mediated quinolone resistance qnrB5 (2/116). Conclusions: Importantly, mobility/context inferences are restricted to this ESBL-screen-enriched WGS subset. Swine-associated E. coli from Hungarian large-scale farms harbors complex resistance architectures shaped by co-selection of mobile ARG modules on top of a pervasive chromosomal resistance backbone. Mobility-aware surveillance and stewardship are warranted to mitigate dissemination risks at the animal–environment–human interface. Full article

This study evaluated the efficiency and ecotoxicological impact of the Fenton oxidation process with different metal-based catalysts (FeSO₄, CuSO₄, MnSO₄) in removing pharmaceuticals and organic contaminants from real hospital wastewater. All catalytic systems achieved high oxidation, with COD reduction reaching 81–89% after 4 h. Two complementary approaches were applied: targeted LC-MS/MS quantification of a model mixture of antibiotics and pharmaceuticals, and untargeted GC-MS/MS screening method for assessing the overall organic contaminant profile. Toxicity was assessed using Microtox^®. Targeted analysis showed complete or near-complete degradation of β-lactams, tetracyclines and most sulfonamides, with slightly lower removal for sulfamethoxazole in FeSO₄ system (96%). Fluoroquinolones and selected pharmaceuticals, such as caffeine and propranolol were more resistant, particularly with CuSO₄ and MnSO₄ catalysts. The untargeted GC-MS/MS screening revealed the highest overall reduction in chromatographic peak areas for FeSO₄ (70%), followed by MnSO₄ (39%) and CuSO₄ (36%). GC-MS/MS profiling confirmed that the Fe-catalyzed process was the most effective in reducing the total chromatographic peak area (70%). However, ecotoxicological assays revealed a significant increase in toxicity post-treatment, with growth inhibition of Allivibrio fischeri reaching 98%. This suggests that high oxidation does not directly correlate with biological safety, likely due to the presence of unconsumed reagents or the formation of transformation products with higher acute toxicity. These findings emphasize the necessity of integrating bioassays into treatment evaluation protocols to assess the true environmental risk of treated effluents. Full article