Search Results (150)

Monitoring of antimicrobial resistance (AMR) in the environment is needed, but standardized methods are lacking. This study evaluated a cost-efficient, kit-based approach for detecting antibiotic-resistant Escherichia coli (AR-E. coli) and predicting multidrug-resistant E. coli (MDR-E. coli) in concentrated animal feeding operations (CAFO)-impacted surface waters in Tulare, California, and Clayton, Michigan. Eighteen Tulare and six Clayton samples were analyzed using modified IDEXX Colilert-18 kits with selective antibiotics. In Round 1 of the study, 702 isolates were tested against 12 antibiotics using disk diffusion to identify ideal additions. No and low resistance to AMP was seen at the unimpacted and less-impacted sites, respectively, while up to 34% of isolates at impacted sites were resistant to AMP. The percentage of isolates resistant to erythromycin was 16% or lower at less-impacted sites, but it ranged up to 64% at impacted sites. In Round 2, 1002 isolates (1704 total) were characterized to compare modified IDEXX results with culture-based results. Resistance to ampicillin (AMP-E. coli-col) detected by IDEXX strongly correlated with MDR prevalence in Clayton (R² = 0.71, p = 0.00197), while extended-spectrum beta-lactamase-producing E. coli (ESBL-E. coli-col) correlated with MDR prevalence in Tulare (R² = 0.70, p < 0.0001). This observation appears to be site-specific. Findings suggest both IDEXX- and culture-based methods can serve as AMR screening tools at CAFO-impacted sites. This work supports community-based monitoring and informs standardized approaches for tracking AMR in inland waters. Full article

Background/Objectives: Antimicrobial resistance (AMR) threatens global public health. This systematic review and meta-analysis, as part of the “ENVIRE” project (interventions to control the dynamics of antimicrobial resistance from chickens through the environment), assesses the prevalence of phenotypic and genotypic resistance, including extended-spectrum beta-lactamases (ESBLs), AmpC beta-lactamases, carbapenemases, colistin, and fluoroquinolone resistance, in broiler chickens and their environment. Methods: The analysis covers the years 2002–2022, focusing on Escherichia (E.) coli, Klebsiella spp., Enterobacter spp., and Citrobacter spp. in fecal, meat, environmental, and other-than-feces samples from observational studies published in PubMed and Web of Science. Quality assessment was performed using the Alberta Heritage Foundation criteria. Results: Data from 170 studies, conducted in Europe, North Africa, and North America, were included. The most frequently studied resistance was to beta-lactam, with focus on ESBL-producing and AmpC beta-lactamase isolates. The pooled prevalence of ESBL-resistant E. coli observed in meat samples at 41% and in fecal samples at 38% demonstrated significant heterogeneity between the studies. The negative binomial regression analysis of prevalence data revealed significantly higher ESBL-producing E. coli rates in European meat samples compared to North African samples. Conclusions: This systematic review revealed substantial variation in prevalence and emphasizes the need for standardized surveillance systems and robust study designs. Full article

Urinary tract infections (UTIs) are a global health concern, with over 150 million cases annually, primarily caused by Escherichia coli. Due to anatomical differences, females, especially children and postmenopausal women, are four times more susceptible. Crucially, E. coli has developed widespread antimicrobial resistance (AMR), including resistance to broad-spectrum agents and the emergence of Extended-Spectrum Beta-Lactamase (ESBL)-producing strains. This retrospective study analyzed hospital records from 95 female patients with positive urine cultures at Siblin Governmental Hospital in 2024. Patients were stratified into three age categories: children (≤18 years), adults (18–64 years) and elderly patients (>64 years). Statistical analysis using SPSS focused on descriptive resistance patterns and differences across age groups. Overall, cephalothin (85.7%) and cefaclor (78.49%) exhibited the highest resistance rates. Conversely, tigecycline (97.22%) and ertapenem (91.67%) showed the highest susceptibility. Resistance patterns varied significantly by age. For instance, elderly patients showed high resistance to agents like Augmentin (52.5%) and cefixime (66.1%), while the pediatric group (≤18 years) displayed exceptionally high resistance to cefixime (90.0%). E. coli isolates show high resistance to conventionally used antibiotics, complicating UTI treatment. These findings highlight the need for continuous local surveillance, particularly focusing on third-generation cephalosporins and beta-lactamase production. Ultimately, age is a critical factor that must be considered when determining empirical antibiotic therapy for UTIs. Full article

Background: The increasing attention on extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli strains isolated from poultry flocks stems from concerns about their virulence potential and zoonotic risk. Of particular significance is the identification of extraintestinal pathogenic E. coli (ExPEC) pathotypes in poultry, as these strains pose not only animal health concerns but also serious threats to food safety and public health. Mapping the genetic background of pathogenicity and antimicrobial resistance is essential for risk assessment and the development of effective control strategies. Methods: A total of 87 E. coli isolates were isolated from tracheal and cloacal swab samples collected from healthy chickens between 2022 and 2023. Whole-genome sequencing was performed using Illumina and MGI next-generation sequencing platforms. Bioinformatic analyses were conducted to identify virulence-associated genes and pathotype markers using multiple reference databases, including VirulenceFinder. The frequency of virulence genes was summarized both in tabular form and visualized through graphical representations. Results: A substantial proportion of the isolates harbored virulence genes linked to various ExPEC pathotypes, particularly uropathogenic E. coli (UPEC), avian pathogenic E. coli (APEC), and neonatal meningitis-causing E. coli (NMEC). The most frequently detected colonization factors included members of the fim, pap, ecp, and fae gene families. Among fitness-related genes, iron acquisition systems—ent, chu, iro, iuc, fep, and ybt—were especially prevalent. Classic UPEC-associated genes such as pap and fimH, along with the APEC-related iutA and vat, were found at high frequencies. Four isolates exhibited a virulence gene profile characteristic of the NMEC pathotype (ibeA, kpsD/M/T, fimH). In contrast, hallmark genes of enteric pathotypes were absent from all isolates. Conclusions: The predominance of extraintestinal virulence factors in the examined poultry-derived E. coli strains underscores their zoonotic potential. The complete absence of enteric pathotype markers indicates that the studied poultry populations primarily harbor ExPEC-like strains. These findings highlight the critical need for ongoing genomic surveillance and targeted preventive strategies within poultry production systems. Full article

Extended-spectrum beta-lactamase (ESBL)-producing E. coli from broiler farms can reach watersheds used for recreational swimming. We assessed short-term swimmer exposure by extending a modular quantitative microbial risk assessment (QMRA) to the recreational water pathway linking land manure application to in-stream fate and transport with dilution and decay. We modeled single-event exposure doses and estimated loss of disability-adjusted life years (DALYs). We ran sensitivity analyses on several parameters and compared outputs to published recreational water assessments that include ESBL E. coli. Assuming a worst-case scenario, single-event doses were lower for adults (2.95 CFU; UI 0.14–6.11) and higher for children (8.78 CFU; UI 0.56–17.20) on day 1 after land application, then dropped below 0.01 CFU by day 200, with DALY losses from 10⁻⁷ to 10⁻¹⁰. Uncertainty was dominated by fate and transport. Stronger particle binding, faster in-stream decay, and larger effective volumes lowered exposure, while higher shedding, greater flow, and larger wash-off raised it. Estimates fell at the low end of prior studies. Swimmer exposure appears to be extremely low and short-lived. The modular QMRA links farm contamination to bathing-site risk and supports risk-based monitoring (after spreading or storms) and short-term forecasts that focus advisories on short, higher-risk windows after litter application. Full article

The emergence of antimicrobial resistance (AMR) across the broiler production chain holds significant economic, animal, and public health implications. This study investigated phenotypic resistance to 13 antimicrobials and the presence of 35 antimicrobial resistance genes (ARGs) in Escherichia coli (n = 291) isolated across three broiler production chains (broiler breeder farms, hatcheries, commercial broiler farms, and retail meat shops). An extremely high phenotypic resistance (>70%) to doxycycline, ciprofloxacin, and cefpodoxime, and very high resistance (50–70%) to ampicillin, cefotaxime, gentamicin, and ceftazidime was observed. In addition, 97% of isolates were multidrug-resistant (resistant to ≥1 drug in ≥3 antimicrobial classes), 42% were extended-spectrum beta-lactamase (ESBL) producers, 65% were resistant to third-generation cephalosporins (3GCR), and 21% were resistant to colistin. The Poisson regression model revealed no significant difference in AMR among broiler production stages, except for colistin. Among 35 ARGs tested, 24 (67%) were detected at least once. The most prevalent were tetA, bla_TEM, qnrB, qnrS, and aac(6′)-Ib-cr, while qnrD, sul2, bla_OXA, and bla_CTX-M were detected at lower levels (1–7%). All five tested mcr genes (mcr-1 to mcr-5) were identified in commercial farms and retail shops. No bla_NDM, tetB, tetC, tetD, tetM, qnrC, aac(3)-IIa (aacC2), aph(3)-IIa (aphA2), or aac(6′)-Ib genes were found. A strong correlation was observed between AMR phenotypes and ARGs. High AMR among E. coli in broiler production poses significant One Health risks, with widespread MDR, ESBL production, and resistance to critically important antimicrobials. Prudent antimicrobial use, enhanced surveillance and education, farm biosecurity, and One Health strategies are crucial in mitigating these threats. Full article

Background/Objectives: Urinary tract infections (UTIs) caused by extended-spectrum beta-lactamase (ESBL) Enterobacterales have become more frequent. Therefore, strategies for assessing the risks posed by ESBL-producing infections have been developed, creating the need for local validation. The aim of this study was to validate the scoring system designed by Tumbarello et al. to identify ESBL producers in patients with a UTI that require hospital care in a region with a high prevalence of ESBL Escherichia coli. Methods: A retrospective cohort study was conducted in a third-level hospital in Bogotá (Colombia) between 2013 and 2020.The study included 817 patients, who were hospitalized due to pyelonephritis and treated with cefuroxime (the first-line antibiotic according to local guidelines), with an ESBL frequency of 9.68%. Diagnostic performances were estimated for a modified version of Tumbarello’s score (omitting admission from another healthcare facility) evaluating the area under the curve (AUC) for ESBL presence with respect to resistance to second- and third-generation cephalosporins. Results: With an index cut-off of ≥6, the score showed a sensitivity of 18% and a specificity of 83%. The AUC for this cut-off was 0.47. This threshold index could not efficiently predict either third- (AUC = 0.49) or second-generation cephalosporin resistance (AUC = 0.51). Conclusions: In Colombia, a region with a high prevalence of ESBL E. coli producers, as the use of the Tumbarello index would result in excessive utilization of wide-spectrum antibiotics, it is not recommended in this specific scenario for UTIs. Further studies are required in order to develop accurate tools to assess the risk of ESBL producers in high-prevalence settings. Full article

Background/Objectives: Extended-Spectrum Beta-Lactamase-producing Escherichia coli (ESBL-E. coli) are widely circulating in livestock of low- and middle-income countries. However, the drivers of their circulation remain largely unknown. Small-scale farms in Peru exhibit an unusually high prevalence of fecal carriage of ESBL-E. coli in their livestock. The objective of this study was to compare the prevalence of ESBL-E. coli fecal carriage in dairy cows, pigs, and poultry in the Lima and Ica regions of Peru and to identify the drivers associated with the observed prevalence at the farm level. Methods: We compared the prevalence of fecal carriage of ESBL-E. coli isolated from dairy cattle (N = 244 animals; 25 farms), pigs (N = 261; 25), and laying hens (N = 255; 10). We also administrated questionnaires to 59 farmers regarding their socioeconomic status, husbandry practices, animal diseases, and antibiotic use. Results: All but one of the 60 farms sampled had at least one animal carrying ESBL-E. coli. A statistically higher prevalence of ESBL-E. coli was estimated in dairy cows (75%) and pigs (61%) from Lima compared to laying hens from Ica (34%). Our statistical analyses (Poisson generalized linear models) using two variable selection approaches revealed that the prevalence of ESBL-E. coli was lower in farms raising laying hens, when farmers oversaw both animal husbandry and healthcare, and in farms with a lower number of gastrointestinal outbreaks in the last semester. Socio-economic features of farmers and self-reporting antibiotic use varied across farms (i.e., highest antibiotic use over the last semester was reported among pig farmers (96%), followed by laying hen (70%) and dairy cattle farmers (50%)), but these factors were not associated with the prevalence of ESBL-E. coli. Conclusions: Despite a relatively low number of farms sampled, our findings suggest that the widespread circulation of ESBL-E. coli among livestock in Peru could be mainly associated with unknown species-specific drivers, independently of the socioeconomic status of farmers and antibiotic use. Therefore, our study calls for future research to identify the specific drivers promoting the high prevalence of ESBL-E. coli among cattle and pigs in Peru. Full article

The emergence and dissemination of extended-spectrum beta-lactamase (ESBL)-producing bacteria pose a major public health threat, necessitating a One Health approach to addressing this threat. Thus, the diversity, ESBL production, and potential public health implications of Gram-negative bacteria recovered from man-made lakes and surrounding lettuce in Yamoussoukro, Côte d’Ivoire were assessed in this study. Also, the lakes’ physicochemical parameters were assessed and correlated with bacteria community using Pearson correlation. A total of 68 Gram-negative bacterial isolates were recovered from the samples and identified via 16S rDNA gene sequencing. Phylogenetic analysis suggested multiple genus-/species-level variations within the isolates. Escherichia coli was the most prevalent in lake water (39.5%), while Acinetobacter was the dominant genus in lettuce (30%). E. coli isolates showed high resistance to ampicillin (90.9%), cefepime (72.7%), cefotaxime (68.2%), and aztreonam (63.6%). Moreover, ESBL production was confirmed in E. coli isolates (22.05%), predominantly mediated by the bla_CTX-M gene. Multidrug-resistant phenotypes were widespread, yielding similar multiple antibiotic resistance index (MARI) values in water (0.27–0.63) and lettuce (0.27–0.81). These data indicate high environmental contamination, which unfortunately is not being taken into account by lettuce producers according to an interview. Statistical analyses showed a significant relationship between bacterial diversity and lakes’ physicochemical parameters, including dissolved oxygen, pH, and turbidity. The basic education level of farmers, the prevalence of ESBL-producing E. coli, and the high prevalence of MDR Gram-negative bacteria in both environmental and crop sources in Yamoussoukro underscore the need for both integrated surveillance and management strategies to mitigate potential microbial public health risks within a One Health framework. Full article

Extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae pose a significant public health risk. As zoos grow in popularity, exotic animals come into closer contact with humans, making them potential reservoirs of ESBLs. However, data on ESBL presence in Mexican zoos remains limited. For this reason, this study aimed to isolate and assess the antimicrobial susceptibility of Enterobacteriaceae that colonize avian species in a petting zoo and to identify any ESBL-producing isolates. Cloacal swabs were collected from 34 healthy birds at Miguel Angel de Quevedo Zoo, Veracruz, Mexico. Samples were analyzed microbiologically and molecularly to detect ESBL-encoding genes. A total of seventeen E. coli and one K. pneumoniae strains were isolated from cloacal swabs of bird species, and multidrug resistance (MDR) was found. The most frequently detected genes were bla_CTX-M-1 (16/18) and bla_TEM-1 (12/18). The detection of multidrug-resistant (MDR) strains carrying bla_CTX-M-1, bla_TEM-1, and bla_SHV genes highlights the potential role of birds as reservoirs and disseminators of antimicrobial resistance (AMR) in urban environments. To the best of our knowledge, this is the first study conducted in Mexico. In conclusion, MDR ESBL-producing bacteria were found in the fecal microbiota of bird species at a petting zoo in Mexico. The limitations of this study emphasize the need for a One Health approach to analyze the genome-wide isolates and epidemiology of antimicrobial-resistant bacteria in captive zoo animals in Mexico. This would support targeted surveillance efforts and help reduce the emergence and spread of resistant bacteria among zoo animals and visitors. Full article

Background: Urinary tract infections (UTIs) represent a significant public health concern, particularly in children with structural abnormalities such as vesicoureteric reflux. Prolonged antibiotic exposure in these patients often contributes to the emergence of multidrug-resistant organisms and restricts therapeutic options. Probiotics have emerged as a potential adjuvant strategy to reduce infection recurrence. Case Presentation: A female infant born at term (38.6 weeks), with a prenatal diagnosis of bilateral hydronephrosis, experienced recurrent UTIs from the neonatal period despite both prophylactic and therapeutic antibiotic regimens. Serial urine cultures revealed infections caused by extended-spectrum beta-lactamase (ESBL)-producing E. coli and K. pneumoniae. Methods: The isolated strains were evaluated in vitro against Lactiplantibacillus sp. LH01, a probiotic strain derived from human milk. Following confirmation of its antimicrobial activity, an individualised intervention was initiated: daily oral administration of 1 mL of the probiotic (10⁹ CFU/mL) for one month, under medical supervision and without concurrent antibiotic therapy. Results: The probiotic demonstrated 89% inhibition efficiency against multidrug-resistant strains, accompanied by a notable reduction in UTIs frequency. Follow-up cultures showed reduced pathogen growth and a loss of the ESBL phenotype, facilitating clinical management and allowing previously contraindicated surgical interventions. Conclusions: Lactiplantibacillus sp. LH01 proved a safe and effective adjuvant in managing recurrent, resistant UTIs in a paediatric patient, highlighting the promise of probiotic therapies. Full article

Among synanthropic species, European hedgehogs are widely distributed throughout Europe. In recent decades, these animals have increasingly adapted to anthropogenic environments, where they find abundant shelter and food resources, along with fewer natural predators. As with other wildlife, it is likely that their coexistence in cities is also affecting their microbiota, promoting the development of antimicrobial resistance (AMR). This study aimed to assess the occurrence and patterns of AMR in commensal enteric Escherichia coli isolated from hedgehogs (n = 53) living in anthropogenic environments upon admission to a wildlife rescue center in Turin (Italy). The effects of hospitalization on the prevalence and trends of AMR were also assessed. Our results confirm that hedgehogs can harbor resistant E. coli upon admission, in particular against cefazolin (41.5%), ampicillin (37.7%), and enrofloxacin (22.6%). In addition, hospitalization promoted an increase in minimum inhibitory concentration (MIC) values of all antibiotics except imipenem, which led to a significant increase in E. coli that was resistant towards doxycycline, enrofloxacin, and trimethoprim-sulfamethoxazole. Admitted hedgehogs were also carriers of extended-spectrum beta-lactamase-producing E. coli (5.7%), whose presence increased during hospitalization (to 20.8%). These results highlight the role of hospitalizations longer than five days in the acquisition of AMR and suggest that European hedgehogs can become potential carriers of resistant E. coli following hospitalization. Full article

Antibiotics are frequently used in the poultry industry, which has led to the emergence of bacterial strains that are resistant to antimicrobial treatments. The main objectives of this research were to conduct a multimodal risk assessment, to determine the extent of contamination of chicken meat with Escherichia coli, assess the prevalence of strains resistant to extended-spectrum cephalosporins (ESC), and characterise the genes associated with resistance and virulence. A standardised procedure involving enrichment in buffered peptone water and isolation of E. coli on MacConkey agar was carried out on 100 chicken carcasses. Subsequently, the sensitivity of the strains was tested against 21 antibiotic discs. Additionally, ESBL production was detected using a double synergy test. Specific PCRs were employed to identify resistance to critical antibiotics in human medicine (such as cephalosporins, carbapenems, fluoroquinolones, and colistin), as well as the presence of virulence genes. The contamination rate of chicken meat with E. coli was 82%. The prevalence of ESC-resistant isolates was 91.2%. Furthermore, 76.5% of the isolates exhibited ESBL production, with the different beta-lactamase genes (bla_CTXM, bla_TEM, and bla_SHV). The mcr-1 gene, associated with colistin resistance, was detected in four strains (5.9%). Some isolates also carried resistance genes such as sul1, sul2, sul3, tetA, tetB, qnrB, and qnrS. In addition, several virulence genes were detected. In our study, we were able to link the expression of AMR to the iron metabolic regulatory elements using a multimodal machine learning approach; this mechanism could be targeted to mitigate the bacteria virulence and resistance. The high prevalence of ESBL-producing and multi-resistant E. coli strains in poultry presents significant human health risks, with the focus on antibiotic-resistant uropathogenic strains since poultry meat could be an important source of uropathogenic strains, underscoring the danger of hard-to-treat urinary tract infections, stressing the need for controlled antibiotic use and thorough monitoring. Full article

Background: Urinary tract infections (UTIs) caused by the multidrug resistance (MDR) phenotype termed extended-spectrum beta lactamase (ESBL)-producing E. coli is a significant and growing global health concern. In response to the rising prevalence, the novel Beta Lactam-Beta Lactamase inhibitor (BL/BLI) combinations have been introduced in recent years. While these agents have shown efficacy, their clinical utility is constrained by high cost, limited availability, and emerging resistance mechanisms. The rational of this study was to test the in vitro activity of a cost-effective alternative to currently available BL–BLI combinations against ESBL-producing E. coli isolated from urinary tract infections (UTIs). Objective: This study investigates the in vitro antimicrobial activity of cefaclor (CFC), both as monotherapy and in combination with the β-lactamase inhibitors clavulanic acid (CA) and sulbactam (SUL), against 52 ESBL-producing E. coli isolates derived from urine cultures of patients diagnosed with UTIs. Methods: The susceptibility ranges were measured by disk diffusion and minimal inhibitory concentration (MIC) methods. In addition, the Time kill assay and disk approximation method were performed to measure the synergistic and bactericidal activity of the approached combination. Results: The MIC50 and MIC90 for CFC were improved from more than 128 µg/mL to 8/4 µg/mL when CFC was combined with either CA or SUL. The triple combination format of CFC/CA/SUL showed MIC50 and MIC90 values at 8/4/4 µg/mL and 64/32/32 µg/mL, respectively. The recovered susceptibility percentages were 54%, 54%, and 58% for CFC/CA, CFC/SUL, and CFC/CA/SUL combinations, respectively. Disk approximation and time–kill assay results revealed synergy and bactericidal effects when CFC combined with CA or SUL for isolates that showed susceptibility restorations of CFC when coupled with CA or SUL by the disk diffusion and MIC method. Conclusions: This study proposes a cost-effective combination that could mitigate resistance development and offer a sparing option to last resort treatment choices including carbapenems. However, testing efficacy in a clinical setting is crucial. Full article

Acute diarrhoeal disease caused by antibiotic-resistant diarrhoeagenic bacteria is a significant global public health issue, particularly in low- and middle-income countries. This study provides the first molecular characterisation of antimicrobial resistance profiles, including the detection of CTX-M-15 and CTX-M-55 extended-spectrum beta-lactamases (ESBLs), among diarrhoeagenic Enterobacterales in Bioko Island, Equatorial Guinea, offering novel epidemiological insights into an understudied region. This study investigated the antibiotic resistance profiles of pathogenic bacteria isolated from diarrhoeal samples on Bioko Island. A total of 153 clinical isolates were collected between 1 February and 30 May 2014, and antimicrobial susceptibility testing was performed at Loeri Comba Polyclinic (Malabo) using the Kirby–Bauer method. The molecular characterisation of β-lactamase-associated genes was performed on different isolates of diarrhoeagenic pathotypes—144 Escherichia coli, 7 Salmonella enterica, and 2 Shigella flexneri—at the National Centre for Microbiology (Majadahonda, Spain). High resistance rates were detected against ampicillin (98%), tetracycline (93.5%), sulfonamides (94.8%), sulfamethoxazole–trimethoprim (88.2%), and cefotaxime (78.8%), while moderate rates of resistance were noted for ciprofloxacin (26.7%), and all isolates remained susceptible to imipenem. Of the isolates, 107 (69.9%) produced either single or multiple β-lactamases. Among these, 73 (68.2%) harbored classical β-lactamases, specifically TEM and OXA-1 types, representing 47.7% of the total sample. Additionally, 34 (31.8%) of the isolates were identified as producers of extended-spectrum β-lactamases (ESBLs), specifically CTX-M enzymes. Sequencing identified CTX-M-15 and CTX-M-55 variants. The predominant ESBL-producing bacteria were enteroaggregative Escherichia coli (56.2%), followed by enteropathogenic and enterotoxigenic E. coli. These findings confirm the circulation of multidrug-resistant diarrhoeagenic Enterobacterales in Equatorial Guinea, raising concerns about limited treatment options due to widespread resistance to multiple antibiotic classes, including third-generation cephalosporins and quinolones. The most important conclusion drawn from this study is that a high percentage of diarrhoeagenic bacteria have an antibiotic resistance and multi-resistance profile, especially to beta-lactams and other groups of antibiotics such as tetracyclines and sulphonamides. There is also a moderate prevalence of isolates carrying ESBLs on Bioko Island, Equatorial Guinea, which could indicate the inappropriate use of antimicrobials. Full article