Science Page: Salmonella monophasic harboring plasmid mediated resistance genes to enrofloxacin and ceftiofur is expanding in swine in the Midwest

This is our Friday rubric: every week a new Science Page from the Bob Morrison’s Swine Health Monitoring Project. The previous editions of the science page are available on our website.

This week, we are sharing a report by Dr. Elnekave and the STEMMA lab regarding a multiresistant clad of Salmonella isolated in the Midwest.

Key Points:

  • A genetically distinct clade of Salmonella 4,[5],12:i:- (also referred to as S. monophasic), harboring multiple antimicrobial resistance genes (including to ampicillin,streptomycin, sulfonamides, and tetracyclines) became the predominant S. monophasic type in swine in the U.S. during 2014-2016.
  • Phenotypic resistance to enrofloxacin (fluoroquinolone) and ceftiofur (3rd generation cephalosporin) was present in a proportion these isolates, and whole genome sequencing revealed the presence of the plasmid-mediated genes.
  • These plasmid-mediated resistance genes could potentially transfer horizontally to other microorganisms and augment the problem of antimicrobial resistance to these critically important antibiotics.

S. monophasic emerged globally in the recent years and pig products have been identified as a source in some foodborne outbreaks. The prevalence of S. monophasic, and phenotypic resistance (minimum inhibitory concentration (MIC) above the cut-off value for this bacteria) to enrofloxacin increased in swine clinical samples in the Midwest during 2006 and 2016.

During this period, injectable enrofloxacin was approved by the Food and Drug Administration (FDA) for treatment of swine respiratory disease and colibacillosis in piglets (in 2008 and 2014, respectively); therefore, the objective of the study was to characterize the S. monophasic in swine in the U.S Midwest.

Salmonella genotypic profile
Maximum likelihood tree of S. monophasic collected in the U.S. and Europe during 1991-2016. Tip colors indicate of the period of sample collection: 1991-2009 (red), 2010-2013 (green), 2014-2016 (turquoise) and not available (n.a.; purple). The location of samples collection is indicated by the background color: Europe (red), U.S. (blue) and not available (green). ASSUT= presence of resistance genes against ampicillin, streptomycin, sulfonamides, and tetracyclines. qnr genes – conferring resistance to quinolones.

We used whole genome sequencing to compare S. monophasic isolates collected from livestock in the Midwest with isolates collected from different sources in the U.S. and Europe. We then determined the antimicrobial resistance genotypes and presence of other virulence factors that could help to explain the emergence of this variant.

Salmonella monophasic formed two main genetic clades regardless of source and geographical origin (Figure 1). Most (84%) isolates recovered in the U.S. during 2014-2016, including 50 isolates (out of 51) originating mainly from swine in the Midwest, were part of an emerging clade genotypically resistant to ampicillin, streptomycin, sulphonamides and tetracyclines. In the Midwest samples, phenotypic resistance to enrofloxacin (11 out of 50; 22%) and ceftiofur (9 out of 50; 18%) was found in conjunction with plasmid-mediated resistance genes. This is of particular concern because fluoroquinolones and 3rd generation cephalosporins are often used to treat invasive Salmonella infections in people. Furthermore, because the genes were plasmid borne there is greater likelihood for horizontal transfer of these genes to other bacterial strains.

Vaccination against Lawsonia intracellularis decreases shedding of Salmonella enterica serovar Typhimurium in co-infected pigs changes the host gut microbiome

This is our Friday rubric: every week a new Science Page from the Bob Morrison’s Swine Health Monitoring Project. The previous editions of the science page are available on our website.

This week,  we are sharing the summary of a publication by  Dr. Fernando Leite who recently received his PhD from the University of Minnesota. The full scientific article regarding the effect of the vaccination against Lawsonia intracellularis on the shedding of Salmonella typhimurium and the host microbiome is available on open access in Nature.

Materials and Methods

A total of five treatment groups were used:

  1. challenged with S. Typhimurium alone,
  2. challenged with both S. Typhimurium and L. intracellularis,
  3. challenged with S. Typhimurium and vaccinated against L. intracellularis,
  4. challenged with both S. Typhimurium and L. intracellularis and vaccinated against L. intracellularis
  5. a non-infected control.

Results

The greatest difference in shedding level between groups was found at 7 days post-infection. At this time point, the co-challenged animals from the vaccinated group shed statistically less S. Typhimurium per gram of feces than the animals from the non-vaccinated, co-challenged group. The co-challenged vaccinated group also shed significantly less S. Typhimurium than the singly infected S. Typhimurium group.
L. intracellularis vaccination did not have a significant impact on S. Typhimurium shedding when animals were singly infected with S. Typhimurium.

Leite Ileitis vaccination salmonelle co infection

 

At 7 days post-infection, different treatment groups had significant differences in their microbiome community structure. The co-infected vaccinated group clustered apart from all other treatment groups.

Conclusion

These results indicate that vaccination against L. intracellularis impacts the microbiome and reduces shedding of S. Typhimurium in co-infected animals.

 

A new multidrug resistant Salmonella enterica serotype found in Midwestern swine

Text reproduced from the Center for Infectious Disease Research and Policy (CIDRAP)

A new study by Dr. Julio Alvarez‘s team from the STEMMA laboratory, published  in Clinical infectious Diseases suggests that a Salmonella strain circulating in pigs in the US Midwest is part of an emerging clade from Europe that is resistant to multiple antibiotics and may pose a public health risk.

The strain, Salmonella 4,[5],12:i:-, causes many foodborne disease outbreaks mostly tied to pigs and pork products and is expanding in the United States, according to the report by researchers from Minnesota and the United Kingdom.

The team used whole-genome sequencing to assess the relatedness of 659 S 4,[5],12:i:- isolates and 325 S Typhimurium isolates from various sources and locations in the United States and Europe. They also searched for resistance genes and other virulence factors and, for 50 livestock isolates and 22 human isolates, determined the antimicrobial resistance phenotypes.

The researchers found that the S 4,[5],12:i:- isolates fell into two main clades, regardless of their host or place of origin. Eighty-four percent of the US isolates recovered from 2014 through 2016, including nearly all those from pigs in the Midwest, were part of an emerging clade. This clade carried multiple genetic markers for antimicrobial resistance, including resistance to ampicillin, streptomycin, sulphonamides, and tetracyclines.

In addition, phenotypic (actual) resistance to enrofloxacin and ceftiofur was found in 11 of the 50 tested livestock isolates and 9 of the 22 human isolates. This was accompanied by plasmid-mediated resistance genes.

The authors conclude that S 4,[5],12:i:- strains circulating in Midwestern swine herds “are likely part of an emerging multidrug resistant clade first reported in Europe, and can carry plasmid-mediated resistance genes that may be transmitted horizontally to other bacteria and thus could represent a public-health concern.”

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salmonella enterica multidru resistant Alvarez 2017

Abstract

Background
Salmonella 4,[5],12:i:-, a worldwide emerging pathogen that causes many foodborne outbreaks mostly attributed to pig and pig products, is expanding in the U.S

Methods
Whole genome sequencing was applied to conduct multiple comparisons of 659 S. 4,[5],12:i:- and 325 S. Typhimurium from different sources and locations (i.e. U.S. and Europe) to assess their genetic heterogeneity, with a focus on strains recovered from swine in the U.S. Midwest. In addition, presence of resistance genes and other virulence factors was detected and the antimicrobial resistance phenotype of 50 and 22 isolates of livestock and human origin, respectively, was determined.

Results
The S. 4,5,12:i:- strains formed two main clades regardless of their source and geographical origin. Most (84%) of the U.S. isolates recovered in 2014–2016, including those (50/51) recovered from swine in the U.S. Midwest, were part of an emerging clade. In this clade, multiple genotypic resistance determinants were predominant, including resistance against ampicillin, streptomycin, sulphonamides and tetracyclines (ASSuT). Phenotypic resistance to enrofloxacin (11/50) and ceftiofur (9/50) was found in conjunction with the presence of plasmid-mediated resistance genes (qnrB19/qnrB2/qnrS1 and blaCMY-2/blaSHV-12, respectively). Also, higher similarity was found between S. 4,[5],12:i:- from the emerging clade and S. Typhimurium from Europe than with S. Typhimurium from the U.S.

Conclusions
Salmonella 4,[5],12:i:- currently circulating in swine in the U.S. Midwest are likely part of an emerging multidrug resistant clade first reported in Europe, and can carry plasmid-mediated resistance genes that may be transmitted horizontally to other bacteria and thus could represent a public-health concern.

Science Page: Salmonella antimicrobial resistance and emergence of a new serotype S.4,[5],12:i:-

This is our Friday rubric: every week a new Science Page from the Bob Morrison’s Swine Health Monitoring Project. The previous editions of the science page are available on our website.

Monitoring antimicrobial resistance is a research topic of utmost importance in the swine industry. Dr. Julio Alvarez at the University of Minnesota is leading some of this effort and this week, his team is presenting the latest results regarding Salmonella antimicrobial resistance in the strains isolated by the University of Minnesota – Veterinary Diagnostic Laboratory between the years 2006 and 2015 and the emergence of a new serotype S.4,[5],12:i:-

Key Points

  • Swine is the reservoir most commonly associated with the S.4,[5],12:i: serotype.
  • The prevalence of S. agona and S. 4,[5],12:i:- in isolates of swine origin recovered from clinical samples received at the Minnesota Veterinary Diagnostic Laboratory (MVDL) in 2006-2015 has increased.
  • In these serotypes an increased proportion of isolates were resistant to ceftiofur and enrofloxacin, compared with other serotypes.
  • The increase in the frequency of isolation of the S.4,[5],12:i:- serotype in humans may be paralleled by a similar increase in swine clinical samples received in the MVDL.

The information synthesized in the figure below is the evolution, over the years, of the percentages of Salmonella isolated at the UMN – VDL, belonging to each of other the following serotypes: typhimurium, agona, derby, typhymurium var5, and 4,5,12:i:-. The increase in the proportion of S.4,5,12:i:- can be seen starting back in 2011-2012.

Salmonella antibiotic resistance

Click here to read the full report about Salmonella serotypes isolated at the UMN – VDL

Beneficial but variable effect of vaccination to control Salmonella in pigs

In this meta-analysis conducted by the Center for Veterinary Health Surveillance (Madrid, Spain) in collaboration with Drs. Conrado, Perez, and Alvarez from the University of Minnesota, the efficacy of vaccination to control Salmonella infection in pigs was evaluated. The meta-analysis reviewed a total of 44 studies focusing on Salmonella typhimurium or Salmonella choleraesuis. Included protocols were using either inactivated (killed) or live-attenuated vaccines.

Results showed that both vaccine types had a similar efficacy and that the most successful control strategies among the ones reviewed were using killed vaccines to control Salmonella choleraesuis.

Alvarez swine salmonella vaccine 2017

Abstract: Consumption or handling of improperly processed or cooked pork is considered one of the top sources for foodborne salmonellosis, a common cause of intestinal disease worldwide. Asymptomatic carrier pigs may contaminate pork at slaughtering; therefore, pre-harvest reduction of Salmonella load can contribute to reduce public health risk. Multiple studies have evaluated the impact of vaccination on controlling Salmonella in swine farms, but results are highly variable due to the heterogeneity in vaccines and vaccination protocols. Here, we report the results of an inclusive systematic review and a meta-analysis of the peer-reviewed scientific literature to provide updated knowledge on the potential effectiveness of Salmonella vaccination. A total of 126 articles describing the use of Salmonella vaccines in swine were identified, of which 44 fulfilled the inclusion criteria. Most of the studies (36/44) used live vaccines, and S. Typhimurium and S. Choleraesuis were the predominant serotypes evaluated. Vaccine efficacy was most often measured through bacteriological isolation, and pooled estimates of vaccine efficacy were obtained as the difference in the percentage of positive animals when available. Attenuated and inactivated vaccines had similar efficacy [Risk Difference = − 26.8% (− 33.8, − 19.71) and − 29.5% (− 44.4, − 14.5), respectively]. No serotype effect was observed on the efficacy recorded for attenuated vaccines; however, a higher efficacy of inactivated vaccines against S. Choleraesuis was observed, though in a reduced sample.

Results from the meta-analysis here demonstrate the impact that vaccination may have on the control of Salmonella in swine farms and could help in the design of programs to minimize the risk of transmission of certain serotypes through the food chain.

Link to the full article