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Streptococcus suis (S. suis) is a major pathogen for pigs, causing large economic losses to the swine industry. Moreover, this bacterium has a zoonotic potential, being capable of infecting humans in close contact with pigs or, less frequently, through contact with pork products. Given its importance in both veterinary and public health, S. suis remains a key topic of research. This study explores the genetic characteristics of 154 S. suis isolates obtained from clinical samples collected from pigs between 2018 and 2022. Whole genome sequencing (WGS) allowed a comprehensive analysis of the S. suis population in Spain, including detection of serotype, sequence type (ST), antimicrobial resistance genes, and virulence-associated genes. This approach also explored the vertical transmission of this pathogen through vertically integrated pyramids, as evidenced by associations between grandmother and mother sow farms, and phylogenetic groups, serotypes, and STs. Our analysis revealed that serotype 9 was the most prevalent in our strain collection, predominantly associated with ST123. Notably, the three most significant virulence genes, encoding the extracellular protein factor (EPF), the muramidase-release protein (MRP), and suilysin (SLY), were not consistently present in all clinical isolates. Regarding antimicrobial resistance, no phenotypic resistance was observed to ceftiofur or florfenicol, while observing low resistance to ampicillin (0.6 %) and enrofloxacin (2.6 %), intermediate resistance to penicillin (22.1 %), and high percentage of non-wild-type isolates to trimethoprim-sulfamethoxazole (57.1 %), and doxycycline (96.1 %). The most prevalent antimicrobial resistance genes (ARGs) were tet(O) (85.1 %) and erm(B) (86.4 %), conferring resistance to tetracyclines and macrolides, respectively, although macrolides were not included in the phenotypic testing. Overall, this study provides key epidemiological insights into this significant systemic pathogen within the Spanish swine population. The findings underscore the importance of understanding sample origins, such as grandmother and mother sow farms, to develop an effective antimicrobial stewardship program for managing S. suis-associated diseases., Competing Interests: Declaration of Competing Interest Authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2025 Elsevier B.V. All rights reserved.)

Actinobacillus pleuropneumoniae (APP) causes significant economic losses to the swine industry. Antibiotic treatment can be challenging due to its clinical urgency and the turnover of antimicrobial susceptibility results from the diagnostic laboratory. The aim of this study was to evaluate the vertical transmission of APP within integrated systems as a criterion for optimising antimicrobial treatment in the field, using whole genome sequencing (WGS). Additionally, the genetic variability of Spanish APP isolates has been assessed to decipher antimicrobial resistance (AMR) determinants, toxin presence, serotype, and phenotype/genotype concordance of AMR. A total of 169 isolates from clinical cases of porcine pleuropneumonia with known antimicrobial susceptibility profiles were sequenced. Additionally, 48 NCBI assemblies were included to perform a phylogenetic analysis. Phylogenetic analysis revealed high association between phylogenetic clusters, serotypes, and presence of toxins that are associated within vertically integrated systems by its epidemiological link. Concordance between presence of AMR determinants (genotype) vs in-vitro antimicrobial susceptibility pattern (phenotype) was acceptable for amoxicillin, florfenicol, oxytetracycline, and enrofloxacin using epidemiological cut-off values (ECOFFs), but low concordance was observed for doxycycline and trimethoprim-sulfamethoxazole (T/S). On the other hand, using CLSI clinical breakpoints (CBPs), concordance was acceptable for florfenicol and enrofloxacin and not evaluated for doxycycline, oxytetracycline, trimethoprim-sulfamethoxazole (T/S), and amoxicillin because no CBP are available for them. Finally, WGS has demonstrated the clonality between isolates that shared a common origin (grandmother's farm) and resistance phenotype, suggesting vertical transmission of this pathogen and supporting the use of the epidemiological approach as a good criterion to optimise the antimicrobial use., Competing Interests: Declaration of Competing Interest Authors declare no conflict of interest, (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

A longitudinal study was conducted to investigate the presence of multidrug antimicrobial resistance (multi-AR) in Salmonella enterica in pigs reared under conventional preventative medicine programmes in Spain and the possible association of multi-AR with ceftiofur or tulathromycin treatment during the pre-weaning period. Groups of 7-day-old piglets were treated by intramuscular injection with ceftiofur on four farms (n=40 piglets per farm) and with tulathromycin on another four farms (n=40 piglets per farm). A control group of untreated piglets (n=30 per farm) was present on each farm. Faecal swabs were collected for S. enterica culture prior to treatment, at 2, 7 and 180days post-treatment, and at slaughter. Minimal inhibitory concentrations of 14 antimicrobial agents, pulsed-field gel electrophoresis and detection of resistance genes representing five families of antimicrobial agents were performed. Plasmids carrying cephalosporin resistant (CR) genes were characterised. Sixty-six S. enterica isolates were recovered from five of eight farms. Forty-seven isolates were multi-AR and four contained bla CTX-M genes harboured in conjugative plasmids of the IncI1 family; three of these isolates were recovered before treatment with ceftiofur. The most frequent AR genes detected were tet(A) (51/66, 77%), sul1 (17/66, 26%); tet(B) (15/66, 23%) and qnrB (10/66, 15%). A direct relation between the use of ceftiofur in these conditions and the occurrence of CR S. enterica was not established. However, multi-AR was common, especially for ampicillin, streptomycin, sulphonamides and tetracycline. These antibiotics are used frequently in veterinary medicine in Spain and, therefore, should be used sparingly to minimise the spread of multi-AR., (Copyright © 2018 The Authors. Published by Elsevier Ltd.. All rights reserved.)

In this study, we investigated the frequency of isolates included in the susceptible-dose dependent (SDD) category, according to the Clinical and Laboratory Standards Institute guidelines, carrying bla TEM , bla SHV and bla CTX genes among 92 Klebsiella pneumoniae and 80 Enterobacter cloacae clinical isolates. The presence of one or more extended-spectrum β-lactamases (ESBLs) genes was observed in 64% K. pneumoniae and 69% E. cloacae isolates. Nineteen isolates were included in SDD interpretive category criteria, of which 15 carried ESBL genes (seven K. pneumoniae and eight E. cloacae). Considering the high proportion of ESBL gene-containing isolates included in the SDD category (79%), we recommend that physicians exercise caution in the use of cefepime for treatment of infections caused by these isolates, reducing possible therapeutic failure, particularly in cases of ESBL-producing bacterial strains.

Wildlife is a natural reservoir of Salmonella and Campylobacter, the most important human foodborne pathogens worldwide. Free-living birds have the potential to transport, over large distances, such zoonotic bacteria that may harbor antimicrobial resistance traits. On the northeastern Iberian coast, we assessed the role of Yellow-legged Gulls ( Larus michahellis ) as reservoirs of antimicrobial resistance in Salmonella and thermophilic Campylobacter isolates recovered from gulls at three colonies, with varying degrees of dependence on refuse dumps as food sources. Of the 39 Salmonella isolates we tested, 17 were multiresistant (resistance to three antimicrobial families), with eight being Salmonella enterica serovar Typhimurium. Other clinically relevant Salmonella serovars showing multiresistance included Hadar, Bredeney, and Virchow. Relevant Campylobacter antimicrobial resistances were detected among three Campylobacter jejuni isolates, of which all three showed resistance to nalidixic acid, two were resistant to ciprofloxacin, one was resistant to enrofloxacin, and one was resistant to tetracycline. Our results highlight the importance of free-living gulls with opportunistic feeding habits in the dissemination of enteric pathogens resistant to multiple antimicrobial agents of public health concern.

This study assessed the dynamics of cephalosporin resistant (CR) E. coli populations during the life cycle of pigs treated early in life with ceftiofur or tulathromycin. The study was conducted at eight conventional pig farms; four for each treatment with ceftiofur or tulathromycin. At each farm, 70 7-day-old piglets were divided into two groups: a control group (n = 30) and a treatment group (n = 40). Faecal samples were collected on day 0 and on days 2, 7 and 180 post-treatment. Sows were also sampled on day 0. CR E. coli were selected on MacConkey agar with ceftriaxone. On five farms, 7-day-old piglets excreted CR E. coli before treatment associated with the presence of CR E. coli in sows. The occurrence of CR E. coli positive animals decreased with increasing piglet age. The remaining three farms tested negative for CR E. coli during the study period. Results demonstrated great variability in the frequency of CR E. coli positive animals between farms, independent of treatment. Treatment with ceftiofur resulted in a transitory increase in the counts of CR E. coli after 48 h. However, other risk factors including the presence of CR E. coli in sows and animal age were more important than antimicrobial treatment. Accordingly, intervention strategies targeting sows would likely have a beneficial effect in reducing the occurrence of antimicrobial resistance in primary pig production., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Flies may act as potential vectors for the spread of resistant bacteria to different environments. This study was intended to evaluate the presence of Escherichia coli strains resistant to cephalosporins in flies captured in the areas surrounding five broiler farms. Phenotypic and molecular characterization of the resistant population was performed by different methods: MIC determination, pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (MLST), and phylotyping. The presence of extended-spectrum beta-lactamase (ESBL) genes, their plasmid location, and the mobile genetic elements involved in their mobilization were studied. Additionally, the presence of 35 genes associated with virulence was evaluated. Out of 682 flies captured, 42 yielded ESBL-producing E. coli. Of these isolates, 23 contained bla(CTX-M-1), 18 contained bla(CTX-M-14), and 1 contained bla(CTX-M-9). ESBL genes were associated mainly with the presence of the IncI1 and IncFIB replicons. Additionally, all the strains were multiresistant, and five of them also harbored qnrS. Identical PFGE profiles were found for E. coli isolates obtained from flies at different sampling times, indicating a persistence of the same clones in the farm environment over months. According to their virulence genes, 81% of the isolates were considered avian-pathogenic E. coli (APEC) and 29% were considered extraintestinal pathogenic E. coli (ExPEC). The entrance of flies into broiler houses constitutes a considerable risk for colonization of broilers with multidrug-resistant E. coli. ESBLs in flies reflect the contamination status of the farm environment. Additionally, this study demonstrates the potential contribution of flies to the dissemination of virulence and resistance genes into different ecological niches., (Copyright © 2015, American Society for Microbiology. All Rights Reserved.)

The aim of this study was to evaluate if the treatments with ceftiofur and amoxicillin are risk factors for the emergence of cephalosporin resistant (CR) E. coli in a pig farm during the rearing period. One hundred 7-day-old piglets were divided into two groups, a control (n = 50) group and a group parenterally treated with ceftiofur (n = 50). During the fattening period, both groups were subdivided in two. A second treatment with amoxicillin was administered in feed to two of the four groups, as follows: group 1 (untreated, n = 20), group 2 (treated with amoxicillin, n = 26), group 3 (treated with ceftiofur, n = 20), and group 4 (treated with ceftiofur and amoxicillin, n = 26). During treatment with ceftiofur, fecal samples were collected before treatment (day 0) and at days 2, 7, 14, 21, and 42 posttreatment, whereas with amoxicillin, the sampling was extended 73 days posttreatment. CR E. coli bacteria were selected on MacConkey agar with ceftriaxone (1 mg/liter). Pulsed-field gel electrophoresis (PFGE), MICs of 14 antimicrobials, the presence of cephalosporin resistance genes, and replicon typing of plasmids were analyzed. Both treatments generated an increase in the prevalence of CR E. coli, which was statistically significant in the treated groups. Resistance diminished after treatment. A total of 47 CR E. coli isolates were recovered during the study period; of these, 15 contained blaCTX-M-1, 10 contained blaCTX-M-14, 4 contained blaCTX-M-9, 2 contained blaCTX-M-15, and 5 contained blaSHV-12. The treatment with ceftiofur and amoxicillin was associated with the emergence of CR E. coli during the course of the treatment. However, by the time of finishing, CR E. coli bacteria were not recovered from the animals., (Copyright © 2015, American Society for Microbiology. All Rights Reserved.)

In a study of university students, the percentage nasal carriage of Staphylococcus aureus was 40.8% (102/250). Of the isolates, MIC(50) of methicillin was 0.5 µg/mL and MIC(90) was 1 µg/mL. Six (5.8%) isolates were methicillin-resistant and carried the mecA gene. These results suggest that community-associated methicillin-resistant S. aureus may be spreading in Brazil.

Objectives: To investigate the prevalence of Streptococcus pneumoniae (pneumococci) in the nasopharynx of healthy children enrolled in public day-care centers of the municipality of Umuarama, state of Paraná, Brazil. The susceptibility of the pneumococcal strains to antimicrobial agents was also studied., Methods: Nasopharyngeal specimens from 212 children were collected from April to October 2008. After the specimens were seeded in blood agar and incubated at 37 degrees C for 24-48 hours, the colonies suspected of belonging to S. pneumoniae were identified using alpha-hemolysis, optochin sensitivity, and bile solubility test. Penicillin susceptibility was investigated using the disk diffusion and dilution tests. Susceptibility to the other antimicrobial agents indicated for the treatment of pneumococcal infections was investigated using the disk diffusion test., Results: The prevalence of nasopharyngeal pneumococci was 43.4% (92/212), with higher rates in children between 2 and 5 years old (p = 0.0005). There was no significant difference between sexes. Intermediate and full resistance to penicillin were found in 34.8 (32/92) and 22.8% (21/92) isolates, respectively. Sixty-seven strains (72.8%) were resistant to sulfamethoxazole-trimethoprim, eight (8.7%) were resistant to erythromycin, and six (6.5%) to tetracycline. One strain was resistant to clindamycin (1.1%) and another was resistant to chloramphenicol (1.1%). All strains were sensitive to levofloxacin, ofloxacin, rifampicin, telithromycin, linezolid, and vancomycin. Nine strains were considered multiresistant because they were resistant to three or more classes of antimicrobial agents., Conclusions: The present study detected a high prevalence of healthy children colonized with penicillin-resistant S. pneumoniae strains who may be important reservoirs of this pathogen in the community.