Salmonella Heidelberg is frequently found in poultry and poultry products and is associated with antimicrobial resistance strains and infections and mortality in humans. Whole-genome sequencing is used to monitor and understand epidemiological factors related to antimicrobial resistance. This study aimed to characterize the phenotypic resistance and sequence the whole genome of Salmonella Heidelberg strains isolated from poultry products in Brazil. Fourteen Salmonella Heidelberg strains isolated from whole broiler chicken carcasses and portions in Brazil between 2013 and 2019 were used in this study. Genus confirmation was performed by polymerase chain reaction. The disk diffusion test was conducted to assess the phenotypical antimicrobial susceptibility of the strains. Whole-genome sequencing was carried out to investigate the presence of antimicrobial resistance genes, plasmids, multilocus sequence typing, and virulence-associated genes. A high frequency of phenotypic resistance to cephalosporins, tetracyclines, and sulfonamides was detected. All strains had mutations in gyrA and parC and contained the genes tet(A), fosA7, and sul. The presence of genes originating from Yersinia pathogenicity islands was also detected. This study identified a high frequency of antimicrobial resistance in Salmonella Heidelberg strains from broilers slaughtered in different regions of Brazil, all belonging to the same sequence type (ST15) and associated with multiple resistance and virulence genes. The presence of the Yersinia high-pathogenicity island was detected, indicating potential virulence. These findings highlight the importance of continuously monitoring antimicrobial resistance to control and prevent foodborne infections and maintain the efficacy of treatments for human salmonellosis.

1. This study evaluated the minimal inhibitory concentration (MIC) of ciprofloxacin and the presence of plasmid-mediated quinolone resistance (PMQR) mechanisms in 97 nontyphoidal Salmonella spp. isolated from broilers and carcasses from three different regions in Brazil. The presence of mutations in quinolone resistance determination regions (QRDRS) was investigated in the ciprofloxacin-resistant strain by DNA sequencing. 2. Most of the Salmonella spp. (85.6%) had intermediate resistance to ciprofloxacin and only one isolate was resistant. MIC breakpoints ranged from ≤0.03 to 1 µg/ml and 67.01% of the strains had a MIC of 0.25 µg/ml (n=65). Thirteen strains (13.40%) were susceptible to ciprofloxacin with MIC ≤0.06 µg/ml. The qnrB gene was detected in eight isolates with intermediate resistance and in two susceptible strains. The other PMQR genes, qnrA, qnrC, qnrD, qnrS, qnrVC, aac(6′)-Ib, qepA, oqxAB and mutations in QRDR were not detected in any strain. 3. There was a high frequency of ciprofloxacin intermediate resistant Salmonella from broiler and broiler carcasses from Brazil. The presence of these strains in poultry and derived products poses a risk to public health.

Salmonella spp. is one of the major bacterial causes of foodborne gastroenteritis in humans. The aim of this study was to investigate antimicrobial susceptibility to cephalosporins and quinolones, and to identify the genetic mechanisms related to this resistance in strains of Salmonella spp. Seventy chicken carcass samples were collected from slaughterhouses in the state of Rio de Janeiro, Brazil. The phenotypic profile was detected by the disk-diffusion method and the search for genes encoding betalactamases, and resistance to quinolones was evaluated by PCR. The search for mutations in gyrA and parC was carried out by sequencing these genes. Eleven strains of Salmonella spp. of different serotypes were isolated. All the strains were resistant to at least one of the antimicrobials tested, and 63.64% (7/11) showed resistance to three or more antimicrobials. In the phenotypic test for ESBL production, 36.36% (4/11) of the strains were considered positive. PCR detected the resistance genes bla _CMY-2, qnrB, bla _CTX-M, and bla _TEM. Among the isolates, 45.45% (5/11) simultaneously presented the bla _CTX-M, bla _TEM, qnrB genes and a mutation (Thr-57→Ser) in parC. Point mutations in the parC gene were detected in all the analyzed samples. Genes such as bla _SHV, qnrA, qnrC, qnrD, qnrS, aac(6’)-Ib, qepA, and oqxAB were not detected. The study identified Salmonella spp. resistant to cephalosporins and quinolones, with resistance genes and mutations in parC, highlighting concerns about the adoption of biosecurity measures, responsible use of antimicrobials, and surveillance of resistant strains in the poultry chain.

RESUMO O objetivo deste trabalho foi estudar a prevalência de MG e MS e a filogenia das cepas circulantes, comparando-as com outras já descritas em poedeiras comerciais no Brasil. Foram coletados 140 suabes traqueais de poedeiras comerciais com sinais respiratórios em seis granjas da região centro-oeste de São Paulo. As amostras foram avaliadas por PCR, com posterior sequenciamento e análise filogenética das cepas identificadas. Das 140 amostras, 16,4% foram positivas para MG e 68,6% para MS. Houve diferença significativa nas frequências de MG e MS por granja, segundo o teste G de independência (P<0,05). Todas as cepas identificadas de MG e MS de granjas distintas apresentaram similaridade tanto pela lipoproteína para MG quanto pela região 16s rRNA para MS. Neste estudo, foi possível observar altas prevalências dos agentes estudados, sendo a de MS maior que a de MG. Foi detectada infecção mista por MG e MS em 11,4% das amostras e sabe-se que esses micoplasmas podem agir de forma sinérgica, agravando o quadro respiratório. As cepas circulantes identificadas, pela análise das regiões gênicas da lipoproteína para MG e 16S rRNA para MS, são similares em todas as granjas estudadas.

Broiler chickens and derived products are a key source of Shiga toxin-producing Escherichia coli (STEC) in humans. This pathotype is responsible for causing severe episodes of diarrhea, which can progress to systemic complications. A rapid and accurate diagnosis of the disease, and early treatment of the infection with antimicrobials, can prevent it worsening. However, multidrug-resistant strains have potentially negative implications for treatment success. In this context, the aim of the present study was to isolate and identify multidrug-resistant STEC strains from broiler chickens and carcasses. Of 171 E. coli strains, isolated by conventional microbiological techniques and submitted to Polymerase Chain Reaction (PCR), for detection of stx1 and stx2 genes, 21.05% (36/171) were STEC pathotype, and most of them (66.67% - 24/36) carried both stx1 and eae genes. The multidrug resistance pattern was observed in 75% (27/36) of STEC strains. The presence of STEC in broiler chickens and carcasses reinforces that these sources may act as reservoirs for this pathotype. Multidrug-resistant bacteria contaminating animal products represent a public health issue because of the possibility of spread of multidrug-resistant determinants in the food chain and a higher risk of failure in human treatment when antimicrobials are needed.