Microevolution of monophasic variant of epidemic Salmonella Typhimurium (S. 4,[5],12:i:-)

Salmonella Typhimurium is a major cause of food-borne gastroenteritis. In 2005 a monophasic variant of S. Typhimurium (S. 4,[5],12;i:-) sequence type ST34 and ASSuT antimicrobial resistance profile (AMR) emerged as a worldwide epidemic. Pigs are the main reservoir of this pathogen, pork and pork products represent the predominant vehicles of S. 4,[5],12;i:-. In this study, whole-genome sequencing was used to investigate the population structure and the persistence of S. Typhimurium and S. 4,[5],12;i:- ST34 on Irish pig farms. Additionally, the ability of the isolates to produce biofilms was assessed as this represents an important strategy for environmental survival and persistence. Farm-specific clones persisting between production cycles were identified. No unequivocal association between biofilm formation and persistence was observed; however, epidemic clones formed more biofilms than non-epidemic genotypes suggesting the advantage conferred by this trait. During persistence on farm, microevolution enhancing biofilm formation, but also AMR and heavy metal resistance was observed following the acquisition of IncHI2 plasmids. The phage-encoded virulence factor SopE was sporadically described in S. Typhimurium but was associated with epidemic types. sopE was acquired by S. 4,[5],12;i:- ST34 following lysogenic conversion by the novel prophage mTmV. The findings that the acquisition of sopE occurred multiple, independent times in S. 4,[5],12;i:- ST34 and was concomitant with the emergence of this pathogen in pigs suggested that the acquisition of sopE was positively selected and may have contributed to the success of S. 4,[5],12:i:- ST34. mTmV was virtually absent in S. Typhimurium serovar types despite its ability to infect a variety of S. Typhimurium genotypes, but disseminated to diverse S. enterica serovars, potentially contributing to the emergence of novel pathovariants. Variants of mTmV and novel bacteriophages carrying sopE, which were identified in association with sopE copy number variation, likely promote the dissemination of sopE

The incC sequence is required for R27 plasmid stability

IncHI plasmids account for multiple antimicrobial resistance in Salmonella and other enterobacterial genera. These plasmids are generally very stable in their bacterial hosts. R27 is the archetype of IncHI1 plasmids. A high percentage of the R27-encoded open reading frames (ORFs) (66.7%) do not show similarity to any known ORFs. We performed a deletion analysis of all non-essential R27 DNA sequences to search for hitherto non-identified plasmid functions that might be required for plasmid stability. We report the identification of a short DNA sequence (incC) that is essential for R27 stability. That region contains several repeats (incC repeats), belongs to one of the three-plasmid replicons (R27 FIA-like) and is targeted by the R27 E protein. Deletion of the incC sequence drastically reduces R27 stability both in Escherichia coli and in Salmonella, the effect being more pronounced in this latter species. Interfering with incC-E protein interaction must lead to a reduced IncHI1 plasmid stability, and may represent a new approach to combat antimicrobial resistance

Microevolution of antimicrobial resistance and biofilm formation of Salmonella Typhimurium during persistence on pig farms

Salmonella Typhimurium and its monophasic variant S. 4,[5],12:i:- are the dominant serotypes associated with pigs in many countries. We investigated their population structure on nine farms using whole genome sequencing, and their genotypic and phenotypic variation. The population structure revealed the presence of phylogenetically distinct clades consisting of closely related clones of S. Typhimurium or S. 4,[5],12:i:- on each pig farm, that persisted between production cycles. All the S. 4,[5],12:i:- strains carried the Salmonella genomic island-4 (SGI-4), which confers resistance to heavy metals, and half of the strains contained the mTmV prophage, harbouring the sopE virulence gene. Most clonal groups were highly drug resistant due to the presence of multiple antimicrobial resistance (AMR) genes, and two clades exhibited evidence of recent on-farm plasmid-mediated acquisition of additional AMR genes, including an IncHI2 plasmid. Biofilm formation was highly variable but had a strong phylogenetic signature. Strains capable of forming biofilm with the greatest biomass were from the S. 4,[5],12:i:- and S. Typhimurium DT104 clades, the two dominant pandemic clones found over the last 25 years. On-farm microevolution resulted in enhanced biofilm formation in subsequent production cycle

Whole-genome epidemiology links phage-mediated acquisition of a virulence gene to the clonal expansion of a pandemic Salmonella enterica serovar Typhimurium clone

Epidemic and pandemic clones of bacterial pathogens with distinct characteristics continually emerge, replacing those previously dominant through mechanisms that remain poorly characterized. Here, whole-genome-sequencing-powered epidemiology linked horizontal transfer of a virulence gene, sopE, to the emergence and clonal expansion of a new epidemic Salmonella enterica serovar Typhimurium (S. Typhimurium) clone. The sopE gene is sporadically distributed within the genus Salmonella and rare in S. enterica Typhimurium lineages, but was acquired multiple times during clonal expansion of the currently dominant pandemic monophasic S. Typhimurium sequence type (ST) 34 clone. Ancestral state reconstruction and time-scaled phylogenetic analysis indicated that sopE was not present in the common ancestor of the epidemic clade, but later acquisition resulted in increased clonal expansion of sopE-containing clones that was temporally associated with emergence of the epidemic, consistent with increased fitness. The sopE gene was mainly associated with a temperate bacteriophage mTmV, but recombination with other bacteriophage and apparent horizontal gene transfer of the sopE gene cassette resulted in distribution among at least four mobile genetic elements within the monophasic S. enterica Typhimurium ST34 epidemic clade. The mTmV prophage lysogenic transfer to other S. enterica serovars in vitro was limited, but included the common pig-associated S. enterica Derby (S. Derby). This may explain mTmV in S. Derby co-circulating on farms with monophasic S. Typhimurium ST34, highlighting the potential for further transfer of the sopE virulence gene in nature. We conclude that whole-genome epidemiology pinpoints potential drivers of evolutionary and epidemiological dynamics during pathogen emergence, and identifies targets for subsequent research in epidemiology and bacterial pathogenesis

Which Patients with Chronic Periprosthetic Joint Infection Are Less Suitable to Successful Two Stage Exchange Arthroplasty Surgery? A Retrospective Clinical Trial

Which Patients with Chronic Periprosthetic Joint Infection Are Less Suitable to Successful Two Stage Exchange Arthroplasty Surgery? A Retrospective Clinical Tria

Increased phage resistance through lysogenic conversion accompanying emergence of monophasic Salmonella Typhimurium ST34 pandemic strain

Salmonella enterica serovar Typhimurium (S. Typhimurium) comprises a group of closely related human and animal pathogens that account for a large proportion of all Salmonella infections globally. The epidemiological record of S. Typhimurium in Europe is characterized by successive waves of dominant clones, each prevailing for approximately 10–15 years before replacement. Succession of epidemic clones may represent a moving target for interventions aimed at controlling the spread and impact of this pathogen on human and animal health. Here, we investigate the relationship of phage sensitivity and population structure of S. Typhimurium using data from the Anderson phage typing scheme. We observed greater resistance to phage predation of epidemic clones circulating in livestock over the past decades compared to variants with a restricted host range implicating increased resistance to phage in the emergence of epidemic clones of particular importance to human health. Emergence of monophasic S. Typhimurium ST34, the most recent dominant multidrug-resistant clone, was accompanied by increased resistance to phage predation during clonal expansion, in part by the acquisition of the mTmII prophage that may have contributed to the fitness of the strains that replaced ancestors lacking this prophage

Increased phage resistance through lysogenic conversion accompanying emergence of monophasic Salmonella Typhimurium ST34 pandemic strain

Salmonella enterica serovar Typhimurium ( S. Typhimurium) comprises a group of closely related human and animal pathogens that account for a large proportion of all Salmonella infections globally. The epidemiological record of S. Typhimurium in Europe is characterized by successive waves of dominant clones, each prevailing for approximately 10-15 years before replacement. Succession of epidemic clones may represent a moving target for interventions aimed at controlling the spread and impact of this pathogen on human and animal health. Here, we investigate the relationship of phage sensitivity and population structure of S. Typhimurium using data from the Anderson phage typing scheme. We observed greater resistance to phage predation of epidemic clones circulating in livestock over the past decades compared to variants with a restricted host range implicating increased resistance to phage in the emergence of epidemic clones of particular importance to human health. Emergence of monophasic S. Typhimurium ST34, the most recent dominant multidrug-resistant clone, was accompanied by increased resistance to phage predation during clonal expansion, in part by the acquisition of the mTmII prophage that may have contributed to the fitness of the strains that replaced ancestors lacking this prophage

Effect of packaging and storage conditions on some quality traits of bovine meat

Packaging is considered one of the most interesting technological aspects of food production and is a constantly evolving subject in food production. The type of packaging is important for the quality and safety of the product and for the visual appearance of the product to be immediately evaluated by consumers. The purpose of this study was to investigate the effect of four different types of modified atmosphere packaging (ATM) and vacuum packaging (VP) currently used by a company in central Italy, on the main qualitative characteristics of beef. For these two traditional and two new solutions with reduced environmental impact and compostable were evaluated. For each type of packaging, two different products were analyzed: steaks and hamburgers. The samples, immediately after production, were transported to the laboratory in refrigerated containers. Several parameters (color, pH, water holding capacity, drip loss, and microbiological characteristics) were evaluated at time 0 and after 7 (T7), 14 (T14) and 21 days (T21) of storage in the dark and at refrigeration temperature (+4°C ± 2°C). The results showed that the two types of packaging have very similar effects on the water-retaining capacity of the steaks. More noticeable differences were recorded by the colorimetric analysis: for both steaks and hamburgers, the products packaged in the traditional packaging appeared brighter and redder than those packaged in the new alternatives. The microbiological analysis of the steaks showed higher values in the “new” packaging. The formation of abundant ropy slime was observed in one of the samples in the “new” modified atmosphere package at T21. The results of this study showed that the technological characteristics (in particular, the color) and the microbiological characteristics of the steaks and hamburgers were better in “old” packaging, with a better appearance and a longer shelf life. The results obtained show how the research for eco-sustainable products for packaging must be addressed, taking into account the effect of the materials on the qualitative and hygienic- sanitary characteristics of the meat.https://www.pagepressjournals.org/index.php/ijfsam2023Paraclinical Science