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Salmonella enterica subsp. enterica serovar Enteritidis ( S. Enteritidis) is one of the leading causes of foodborne infections associated with broilers and laying hens. Portugal has had the lowest notification rates of salmonellosis in recent years, due to the vaccinations of layer and breeder flocks and strict compliance with biosecurity measures. However, data about the genetic diversity of S. Enteritidis in Portugal are scarce. In this study, 102 S. Enteritidis isolates selected from human ( n = 63) and non-human sources ( n = 39) were characterized by serotyping, antimicrobial susceptibility, and whole genome sequencing. The S. Enteritidis population was mainly resistant to fluoroquinolones, and a sole isolate showed resistance to extended-spectrum cephalosporins. ST11 was the most frequent sequence type, and three novel STs from human isolates (ST9236, ST4457, and ST9995) were assigned. Several Salmonella pathogenic islands (SPI) and Putative SPI were present in the genomes, namely SPI-1, 2, 3, 4, 5, 9, 10, 12, 13, and 14, C63PI, CS54_island, and 170 virulence genes were identified. The phylogenetic analysis revealed that strains from Portugal are genetically heterogeneous regarding sample type, collection date, and genetic content. This study increases the available data, essential to a better characterization of strains in a global context.

The purpose of this study was (i) to compare two groups (players with more vs. less match play time) regarding body composition, vertical and horizontal jumping performance, and aerobic capacity; and (ii) to test the relationships between physical fitness and play time. This study followed a case-control design in which the outcome was playtime, and the causal attribute was physical fitness. Sixty-six youth male soccer players from under-16 (n = 21), under-17 (n = 19), under-18 (n = 12), and under-19 (n = 14) age groups were monitored for match play time during five months of observation. Inclusion criteria consisted of (1) no absence of more than a week due to injury or other conditions during the five months of observation and (2) physical assessments having been done simultaneously with those of the other players (at the beginning of the season). The exclusion criteria were (1) not participating in one week or more of training sessions, and (2) not participating in the physical fitness assessments. At the beginning of the season, players were assessed for anthropometry (height, body mass, skinfolds), countermovement jump, triple hop bilateral and unilateral jump, and aerobic capacity using the Yo-Yo Intermittent Recovery Test, level 2 (YYIRT). The group that played more time had significantly greater YYIRT results (+28.2%; p = 0.009; Cohen’s d = 0.664). No other significant differences were found between those who played more and fewer minutes. Moderate and significant linear positive correlations were found between YYIRT and play time in the under-19 group (r = 0.423; p = 0.031) and overall (r = 0.401; p < 0.001). In the case of the under-17 group, moderate and significant linear positive correlations were found between TSA and play time (r = 0.473; p = 0.041). This suggests that aerobic and anaerobic capacity is related to play time while jumping performance and fat mass seem not to play an essential role in play time., Competing Interests: The authors declare no conflict of interest.

Livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) ST398 is mainly found in Europe and North America, colonizing the nasal cavity of pigs. This study characterized the MRSA isolates recovered from pig nasal swabs ( n = 171) by evaluating the antimicrobial susceptibility profile by broth microdilution and characterizing the genetic lineages by spa -typing. Three linezolid-resistant isolates were subjected to Whole-Genome Sequencing (WGS). All strains harbored the mecA gene and were resistant to tetracycline and susceptible to vancomycin. A high frequency of multidrug resistance (97.6%) was evidenced, with 55 different multidrug resistance profiles identified. The MRSA strains were found to belong to 17 spa -types, three being novel. The linezolid-resistant strains appeared to belong to the ST398 type, spa -type t011, and SCC mec &#95;type&#95;Vc and to harbor the cfr , fex A, bla Z, mec A, tet M, and tet K genes. The cfr gene was predicted to be carried in the plasmid, flanked by IS Sau9 and the transposon TnpR. MRSA from Portuguese fattening pigs present a high diversity of genetic lineages. The presence of cfr -positive LA-MRSA may represent a risk of transmission to humans, mainly to those in contact with livestock.

The intensive and extensive broiler production systems imply different veterinary interventions, including the use of antimicrobials. This study aimed to compare the antimicrobial susceptibility profiles of Escherichia coli isolated from both systems, characterize resistance mechanisms to β-lactams and polymyxins, and identify genetic elements such as integrons. E. coli isolates recovered from broiler cecal samples were assayed for antimicrobial susceptibility through the broth microdilution technique. The molecular characterization of acquired resistance mechanisms to β-lactams and colistin and the detection of integrons was performed by a multiplex PCR. For most antibiotics tested, the prevalence of reduced susceptibility is higher in commensal and extended-spectrum β-lactamases (ESBL)/AmpC producers from broilers raised in the intensive system, compared with those raised under extensive conditions. SHV-12 was the most common ESBL enzyme found in both production systems. Other ESBL variants such as CTX-M-1, CTX-M-55, CTX-M-14, CTX-M-32, CTX-M-9, TEM-52, and plasmid-encoded AmpC enzyme CMY-2 were also present. MCR-1 was identified in a colistin-resistant isolate from broilers raised under the intensive system. This study highlights the differences in E. coli antibiotic susceptibility from both production types and emphasizes that a great deal of work remains to decrease consumption and antimicrobial resistance levels.

Enterococci are part of the commensal gut microbiota of mammals, with Enterococcus faecalis and Enterococcus faecium being the most clinically relevant species. This study assesses the prevalence and diversity of enterococcal species in cattle ( n = 201) and pig ( n = 249) cecal samples collected in 2017. Antimicrobial susceptibility profiles of E. faecium ( n = 48) and E. faecalis ( n = 84) were assessed by agar and microdilution methods. Resistance genes were screened through PCR and nine strains were analyzed by Whole Genome Sequencing. A wide range of enterococci species was found colonizing the intestines of pigs and cattle. Overall, the prevalence of resistance to critically important antibiotics was low (except for erythromycin), and no glycopeptide-resistant isolates were identified. Two daptomycin-resistant E. faecalis ST58 and ST93 were found. Linezolid-resistant strains of E. faecalis ( n = 3) and E. faecium ( n = 1) were detected. Moreover, oxazolidinone resistance determinants optrA ( n = 8) and poxtA ( n = 2) were found in E. faecalis (ST16, ST58, ST207, ST474, ST1178) and E. faecium (ST22, ST2138). Multiple variants of optrA were found in different genetic contexts, either in the chromosome or plasmids. We highlight the importance of animals as reservoirs of resistance genes to critically important antibiotics.

The present study aimed to characterize the extended-spectrum β-lactamases and plasmid-mediated AmpC β-lactamases (ESBL/PMAβ) among Escherichia coli producers isolated from beef, pork, and poultry meat collected at retail, in Portugal. A total of 638 meat samples were collected and inoculated on selective medium for the search of E. coli resistant to 3rd generation cephalosporins. Isolates were characterized by antimicrobial susceptibility testing, molecular assays targeting ESBL/AmpC, plasmid-mediated quinolone resistance (PMQR), and plasmid-mediated colistin resistance (PMCR) encoding genes. The highest frequency of E. coli non-wild type to 3rd generation cephalosporins and fluoroquinolones was observed in broiler meat (30.3% and 93.3%, respectively). Overall, a diversity of acquired resistance mechanisms, were detected: bla ESBL [ bla CTX-M-1 ( n = 19), bla CTX-M-15 ( n = 4), bla CTX-M-32 ( n = 12), bla CTX-M-55 ( n = 8), bla CTX-M-65 ( n = 4), bla CTX-M-27 ( n = 2), bla CTX-M-9 ( n = 1), bla CTX-M-14 ( n = 11), bla SHV-12 ( n = 27), bla TEM-52 ( n = 1)], bla PMAβ [ bla CMY-2 ( n = 8)], PMQR [ qnrB ( n = 27), qnrS ( n = 21) and aac(6')-Ib-type ( n = 4)] and PMCR [ mcr-1 ( n = 8)]. Our study highlights that consumers may be exposed through the food chain to multidrug-resistant E. coli carrying diverse plasmid-mediated antimicrobial resistance genes, posing a great hazard to food safety and a public health risk.

Persistent infections, usually associated with biofilm-producing bacteria, are challenging for both medical and scientific communities. The potential interest in drug repurposing for biofilm control is growing due to both disinvestment in antibiotic R&D and reduced efficacy of the available panel of antibiotics. In the present study, the antibacterial and antibiofilm activities of four non-steroidal anti-inflammatory drugs (NSAIDs), piroxicam (PXC), diclofenac sodium (DCF), acetylsalicylic acid (ASA) and naproxen sodium (NPX) were evaluated against Escherichia coli and Staphylococcus aureus . The minimum inhibitory/bactericidal concentrations (MICs and MBCs) and the dose-response curves from exposure to the selected NSAIDs were determined. MICs were found for PXC (800 μg/mL) and ASA (1750 μg/mL) against E. coli , and for DCF (2000 μg/mL) and ASA (2000 μg/mL) against S. aureus . No MBCs were found (>2000 μg/mL). The potential of NSAIDs to eradicate preformed biofilms was characterized in terms of biofilm mass, metabolic activity and cell culturability. Additionally, the NSAIDs were tested in combination with kanamycin (KAN) and tetracycline (TET). ASA, DCF and PXC promoted significant reductions in metabolic activity and culturability. However, only PXC promoted biofilm mass removal. Additive interactions were obtained for most of the combinations between NSAIDs and KAN or TET. In general, NSAIDs appear to be a promising strategy to control biofilms as they demonstrated to be more effective than conventional antibiotics.