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Summary: Wild birds are common reservoirs of Salmonella enterica. Wild birds carrying resistant S. enterica may pose a risk to public health as they can spread the resistant bacteria across large spatial scales within a short time. Here, we whole‐genome sequenced 375 S. enterica strains from wild birds collected in 41 U.S. states during 1978–2019 to examine bacterial resistance to antibiotics and heavy metals. We found that Typhimurium was the dominant S. enterica serovar, accounting for 68.3% (256/375) of the bird isolates. Furthermore, the proportions of the isolates identified as multi‐antimicrobial resistant (multi‐AMR: resistant to at least three antimicrobial classes) or multi‐heavy metal resistant (multi‐HMR: resistant to at least three heavy metals) were both 1.87% (7/375). Interestingly, all the multi‐resistant S. enterica (n = 12) were isolated from water birds or raptors; none of them was isolated from songbirds. Plasmid profiling demonstrated that 75% (9/12) of the multi‐resistant strains carried resistance plasmids. Our study indicates that wild birds do not serve as important reservoirs of multi‐resistant S. enterica strains. Nonetheless, continuous surveillance for bacterial resistance in wild birds is necessary because the multi‐resistant isolates identified in this study also showed close genetic relatedness with those from humans and domestic animals. [ABSTRACT FROM AUTHOR]

Antimicrobial food packaging involves packaging the foods with antimicrobials to protect them from harmful microorganisms. In general, antimicrobials can be integrated with packaging materials via direct incorporation of antimicrobial agents into polymers or application of antimicrobial coating onto polymer surfaces. The former option is generally achieved through thermal film‐making technology such as compression molding or film extrusion, which is primarily suitable for heat‐stable antimicrobials. As a nonthermal technology, surface coating is more promising compared to molding or extrusion for manufacturing food packaging containing heat‐sensitive antimicrobials. In addition, it also has advantages over direct incorporation to preserve the packaging materials' bulk properties (e.g., mechanical and physical properties) and minimize the amount of antimicrobials to reach sufficient efficacy. Herein, antimicrobial food packaging films achieved through surface coating is explored and discussed. The two components (i.e., film substrate and antimicrobials) consisting of the antimicrobial‐coated films are reviewed as plastic/biopolymer films; and synthetic/naturally occurring antimicrobials. Furthermore, special emphasis is given to different coating technologies to deposit antimicrobials onto film substrate. Laboratory coating techniques (e.g., knife coating, bar coating, and spray coating) commonly applied in academic research are introduced briefly, and scalable coating methods (i.e., electrospinning/spraying, gravure roll coating, flexography coating) that have the potential to bring laboratory‐developed antimicrobial‐coated films to an industrial level are explained in detail. The migration profile, advantages/drawbacks of antimicrobial‐coated films for food applications, and quantitative analyses of the reviewed antimicrobial‐coated films from different aspects are also covered in this review. A conclusion is made with a discussion of the challenges that remain in bringing the production of antimicrobial‐coated films to an industrial level. [ABSTRACT FROM AUTHOR]

Non‐typhoidal Salmonella (NTS) are a significant source of foodborne illness worldwide, with disease symptoms most often presenting as self‐limiting gastroenteritis; however, occasionally the infection spreads and becomes invasive, frequently requiring anti‐microbial treatment. The cattle‐adapted Dublin serovar of NTS has commonly been associated with invasive illness and anti‐microbial resistance (AMR). Here, the enhanced resolution conferred by whole‐genome sequencing was utilized to elucidate and compare the resistome and genetic relatedness of 14 multidrug‐resistant (MDR) and one pan‐susceptible S. Dublin, isolated primarily in Pennsylvania, from fresh retail meat (one isolate) and humans (14 isolates). Twelve different genetic AMR determinants, including both acquired and chromosomal, were identified. Furthermore, comparative plasmid analysis indicated that AMR was primarily conferred by a putative IncA/C2 plasmid. A single pan‐susceptible S. Dublin isolate, collected from the same timeframe and geographical region as the MDR isolates, did not carry an IncA/C2 replicon sequence within its genome. Moreover, the pan‐susceptible isolate was genetically distinct from its MDR counterparts, as it was separated by ≥267 single nucleotide polymorphisms (SNPs), whereas there was a ≤38 SNP distance between the MDR isolates. Collectively, this data set advances our understanding of the genetic basis of the highly drug‐resistant nature of S. Dublin, a serovar with significant public health implications. [ABSTRACT FROM AUTHOR]

Enteroaggregative Escherichia coli (EAEC) is defined by aggregative adherence (AA) to HEp-2 cells, where bacteria display adherence to cell surfaces and also to the intervening substratum in a stacked-brick configuration. We previously showed that an AraC homologue designated AggR is required for the expression of plasmid-encoded genes that mediate AA of EAEC strain 042. In this study, we hypothesized that AggR also controls the expression of other virulence determinants in EAEC 042. Using proteomic and microarray analysis, we identified for the first time that AggR activates the expression of chromosomal genes, including 25 contiguous genes ( aaiA–Y), which are localized to a 117 kb pathogenicity island (PAI) inserted at pheU. Many of these genes have homologues in other Gram-negative bacteria and were recently proposed to constitute a type VI secretion system (T6SS). AaiC was identified as a secreted protein that has no apparent homologues within GenBank. EAEC strains carrying in-frame deletions of aaiB, aaiG, aaiO or aaiP still synthesized AaiC; however, AaiC secretion was abolished. Cloning of aai genes into E. coli HB101 suggested that aaiA–P are sufficient for AaiC secretion. A second T6SS was identified within the pheU PAI that secretes a protein unrelated by sequence identity to AaiC. Distribution studies indicated that aaiA and aaiC are commonly found in EAEC isolates worldwide, particularly in strains defined as typical EAEC. These data support the hypothesis that AggR is a global regulator of EAEC virulence determinants, and builds on the hypothesis that T6SS is an importance mediator of pathogenesis. [ABSTRACT FROM AUTHOR]

Enteroaggregative Escherichia coli (EAEC) is increasingly recognized as a diarrhoeal pathogen in developing and industrialized countries. Most EAEC virulence factors thus far described are encoded on virulence plasmid pAA, yet recent completion of the EAEC genome has suggested the presence of additional factors encoded on chromosomal islands. Previous reports have recognized the presence of a type III secretion system (T3SS), designated ETT2, at the glyU locus of prototype EAEC strain 042, along with possible T3SS effectors at the selC locus. The selC locus was also noted to harbour homologues of Salmonella enterica regulator HilA and of invasin from Yersinia spp., yet previous publications suggested that these loci may be silent. Here, we show that the genes of the selC locus are present inconsistently among a collection of well-characterized EAEC strains. Notably, however, there was perfect correlation between the presence of hilA-homologue eilA and predicted Yersinia invasin homologue gene eaeX. We hypothesized that if expressed, the putative gene product EilA would contribute to EAEC virulence in part by activation of the T3SS and its effectors. An eilA mutant was constructed in EAEC strain 042, and complementation was achieved by cloning the eilA gene under control of an arabinose-dependent promoter. In this system, we observed expression of at least seven genes to be affected by expression of eilA, either directly or indirectly: selC locus genes eipB, eipC, eipD, eicA and eaeX (renamed here air), as well as glyU ETT2 genes eivF and eivA. Notably, the eilA mutant was shown to be less adherent to epithelial cells in culture and to form less abundant biofilms than the isogenic parent. These effects were recapitulated in the air mutant, suggesting that the predicted outer membrane protein product of the air gene is involved as an accessory adhesin and aggregin of EAEC, coexpressed with the T3SS. Our data suggest that the T3SS of EAEC and presumed effectors located on different chromosomal islands may be coordinately activated by EilA, which also activates the genetically linked high molecular weight bacterial surface protein Air. Contributions of this new putative virulence-related regulon in EAEC may include adherence, aggregation, and as yet uncharacterized roles for the T3SS. [ABSTRACT FROM AUTHOR]