A High-Throughput Short Sequence Typing Scheme for Serratia marcescens Pure Culture and Environmental DNA

Molecular typing methods are used to characterize the relatedness between bacterial isolates involved in infections. These approaches rely mostly on discrete loci or whole genome sequences (WGS) analyses of pure cultures. On the other hand, their application to environmental DNA profiling to evaluate epidemiological relatedness amongst patients and environments has received less attention. We developed a specific, high-throughput short sequence typing (HiSST) method for the opportunistic human pathogen Serratia marcescens . Genes displaying the highest polymorphism were retrieved from the core genome of 60 S. marcescens strains. Bioinformatics analyses showed that use of only three loci (within bssA , gabR and dhaM ) distinguished strains with a high level of efficiency. This HiSST scheme was applied to an epidemiological survey of S. marcescens in a neonatal intensive care unit (NICU). In a first case study, a strain responsible for an outbreak in the NICU was found in a sink drain of this unit, by using HiSST scheme and confirmed by WGS. The HiSST scheme was also applied to environmental DNA extracted from sink-environment samples. Diversity of S. marcescens was modest, with 11, 6 and 4 different sequence types (ST) of gabR , bssA and dhaM loci amongst 19 sink drains, respectively. Epidemiological relationships amongst sinks were inferred on the basis of pairwise comparisons of ST profiles. Further research aimed at relating ST distribution patterns to environmental features encompassing sink location, utilization and microbial diversity is needed to improve the surveillance and management of opportunistic pathogens. Importance Serratia marcescens is an important opportunistic human pathogen, multidrug resistant and often involved in outbreaks of nosocomial infections in neonatal intensive care unit. Here, we propose a quick and user-friendly method to select the best typing scheme for nosocomial outbreaks in relating environmental and clinical sources. This method, named high-throughput short sequence typing (HiSST), allows to distinguish strains and to explore the diversity profile of non-culturable S. marcescens . The application of HiSST profile analysis for environmental DNA offers new possibilities to track opportunistic pathogens, identify their origin and relate their distribution pattern with environmental features encompassing sink location, utilization and microbial diversity. Adaptation of the method to other opportunistic pathogens is expected to improve knowledge regarding their ecology, which of significant interest for epidemiological risk assessment and elaborate outbreak mitigation strategies.

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Development and Validation of a High-throughput Short Sequence Typing Scheme for Serratia marcescens Pure Culture and Environmental DNA

10.1101/2021.07.01.450819 ◽

2021 ◽

Author(s):

Thibault Bourdin ◽

Alizée Monnier ◽

Marie-Ève Benoit ◽

Emilie Bédard ◽

Michèle Prévost ◽

...

Keyword(s):

Serratia Marcescens ◽

High Throughput ◽

Neonatal Intensive Care ◽

Environmental Dna ◽

Distribution Patterns ◽

Epidemiological Survey ◽

Short Sequence ◽

Bioinformatics Analyses ◽

First Case ◽

Pure Cultures

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Severe Acute Infection Due to Serratia marcescens Causing Respiratory Distress in An Immunocompetent Adult

Romanian Journal of Internal Medicine ◽

10.1515/rjim-2016-0013 ◽

2016 ◽

Vol 54 (2) ◽

pp. 134-136 ◽

Cited By ~ 2

Author(s):

Pablo Ruiz-Sada ◽

Mikel Escalante ◽

Eva Lizarralde

Keyword(s):

Risk Factors ◽

Respiratory Distress ◽

Serratia Marcescens ◽

Distress Syndrome ◽

Acute Infection ◽

Human Pathogen ◽

Immunocompetent Adult ◽

Uncommon Presentation ◽

Opportunistic Human Pathogen

AbstractThe role ofSerratia marcescenschanged from a harmless saprophytic microorganism to an important opportunistic human pathogen. It often causes nosocomial device-associated outbreaks and rarely serious invasive community acquired infections. We present a case of a community-acquiredSerratia marcescensbacteremia leading to Respiratory Distress Syndrome in a previously healthy 51-year-old man without identifiable risk factors. Full recovery was achieved with solely medical treatment and observation in ICU during three days. To our knowledge it is an extremely uncommon presentation and just few cases have been previously reported in the literature.

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Serratia marcescens Infections

Infection Control ◽

10.1017/s0195941700064201 ◽

1986 ◽

Vol 7 (5) ◽

pp. 273-280 ◽

Cited By ~ 45

Author(s):

Jacques F. Acar

Keyword(s):

Serratia Marcescens ◽

Clinical Laboratory ◽

Clinical Specimen ◽

Human Pathogen ◽

The Family ◽

Opportunistic Human Pathogen

The recognition of serratia as an opportunistic human pathogen can be dated from 1959, when the microorganism entered the family of Enterobacteriaceae, with features recognizable in the clinical laboratory and related to the Klebsiella/Enterobacter group. Since then, physicians have been challenged to establish the significance of isolation of serratia from a clinical specimen.

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The Protective Role of 1,8-Dihydroxynaphthalene–Melanin on Conidia of the Opportunistic Human Pathogen Aspergillus fumigatus Revisited: No Role in Protection against Hydrogen Peroxide and Superoxides

mSphere ◽

10.1128/msphere.00874-21 ◽

2022 ◽

Author(s):

E. M. Keizer ◽

I. D. Valdes ◽

B. L. McCann ◽

E. M. Bignell ◽

H. A. B. Wösten ◽

...

Keyword(s):

Hydrogen Peroxide ◽

Aspergillus Fumigatus ◽

Protective Role ◽

Oxygen Species ◽

Human Pathogen ◽

Opportunistic Pathogens ◽

Green Pigment ◽

Content Type ◽

Opportunistic Human Pathogen

Opportunistic pathogens like Aspergillus fumigatus have strategies to protect themselves against reactive oxygen species like hydrogen peroxides and superoxides that are produced by immune cells. DHN-melanin is the green pigment on conidia of Aspergillus fumigatus and more than 2 decades ago was reported to protect conidia against hydrogen peroxide.

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Complete Genome Sequence of Serratia marcescens Podophage Parlo

Microbiology Resource Announcements ◽

10.1128/mra.00569-19 ◽

2019 ◽

Vol 8 (27) ◽

Cited By ~ 3

Author(s):

Ryan Bockoven ◽

Jamie Gutierrez ◽

Heather Newkirk ◽

Mei Liu ◽

Jesse Cahill ◽

...

Keyword(s):

Serratia Marcescens ◽

Genome Sequence ◽

Complete Genome Sequence ◽

Resistance Mechanisms ◽

Full Genome Sequence ◽

Multiple Resistance ◽

Full Genome ◽

Human Pathogen ◽

Content Type ◽

Opportunistic Human Pathogen

ABSTRACT Serratia marcescens is an opportunistic human pathogen with multiple resistance mechanisms that infects hospitalized patients. Here, we report the full genome sequence of S. marcescens podophage Parlo. Parlo is most similar to Erwinia phage PEp14 and encodes a 3,764-residue protein assumed to be a homolog of DarB, an antirestriction protein.

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Complete Genome Sequence of Serratia marcescens Siphophage Serbin

Microbiology Resource Announcements ◽

10.1128/mra.00422-19 ◽

2019 ◽

Vol 8 (19) ◽

Cited By ~ 3

Author(s):

Eric A. Williams ◽

Helena Hopson ◽

Andrea Rodriguez ◽

Rohit Kongari ◽

Rachele Bonasera ◽

...

Keyword(s):

Urinary Tract ◽

Serratia Marcescens ◽

Genome Sequence ◽

Complete Genome Sequence ◽

Complete Genome ◽

Human Pathogen ◽

Content Type ◽

Opportunistic Human Pathogen ◽

Tract Infections ◽

Hospital Acquired

Serratia marcescens is an opportunistic human pathogen that is known to cause hospital-acquired respiratory and urinary tract infections. Here, we announce the complete genome sequence and the features of S. marcescens phage Serbin.

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Development of a New High-Throughput Multilocus Sequence Typing Method to Monitor Causative Agents of Nosocomial Infections

Infection Control and Hospital Epidemiology ◽

10.1017/ice.2020.726 ◽

2020 ◽

Vol 41 (S1) ◽

pp. s187-s187

Author(s):

Thibault Bourdin ◽

Emilie Bedard ◽

Marie-Ève Benoit ◽

Michèle Prévost ◽

Etienne Robert ◽

...

Keyword(s):

High Throughput ◽

Nosocomial Infections ◽

In Silico ◽

Multilocus Sequence Typing ◽

Phylogenetic Trees ◽

Culture Media ◽

Environmental Dna ◽

Opportunistic Pathogen ◽

Opportunistic Pathogens ◽

Bioinformatics Analyses

Background: Nosocomial infections cause 4%–56% mortality in newborns. Several epidemiological studies have shown that transmission of opportunistic pathogens from the sink to the patient, including Pseudomonas aeruginosa, Stenotrophomonas maltophilia, and Serratia marcescens are associated with nosocomial infections in neonatal intensive care units (NICUs). In this project, we aimed to develop fast, accurate, and high-throughput multilocus sequence typing assays (HiMLST-Illumina) to detect opportunistic pathogens to assess their distribution in the sink environment of NICUs and their transfer to patients. Methods: Genome sequences of P. aeruginosa (n = 45), S. maltophilia (n = 23) and S. marcescens (n = 34) strains were retrieved from public genome databases to build their pangenomes, using the open-source PGAdb-builder server. The core genome was identified for each opportunistic pathogen and was searched for genes displaying the highest polymorphism. The minimal number of loci to include in a HiMLST-Illumina assay was determined by comparing topology of phylogenetic trees of concatenated loci based on genome similarity, computed as the average nucleotide identity (ANI) score. The primers used for HiMLST-Illumina schemes were designed in silico on a conserved domain and were tested on reference strains of each species. Results: Bioinformatics analyses showed that 3–4 loci (<300 base pairs per locus) distinguished strains with the same performances than ANI scores. The assays were tested using opportunistic pathogen isolates and environmental DNA originating from NICU sinks. The HiMLST-Illumina analysis of environmental DNA revealed the presence of at least 1 of the 3 studied opportunistic pathogens in 50% of sampled drains (n = 20). In a previous sampling, P. aeruginosa was isolated on selective culture media before and 48 hours after disinfection of a sink drain with chlorine. S. marcescens was also isolated from another sink 2 weeks after disinfection. Identification of the isolates was confirmed by HiMLST-Illumina analyses and will be typed to compare with clinical isolates. Conclusions: Initial in silico tests predict a high discriminating power of the HiMLST-Illumina method, suggesting that it would be possible to quickly identify strains of interest in a large number of samples. The power of this method is also in the possibility for molecular typing without a need for cultivation. Preliminary results suggest that sinks are readily colonized by opportunistic pathogens. This HiMLST-Illumina scheme will be applied in a 2-year intensive survey of NICUs in 3 hospitals in Montreal to evaluate the performance of new sink designs in limiting bioaerosol production and transmission of opportunistic pathogens to patients.Funding: NoneDisclosures: None

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The A component (SmhA) of a tripartite pore-forming toxin from Serratia marcescens: expression, purification and crystallographic analysis

Acta Crystallographica Section F Structural Biology Communications ◽

10.1107/s2053230x20013862 ◽

2020 ◽

Vol 76 (12) ◽

pp. 577-582

Author(s):

Alicia M. Churchill-Angus ◽

Svetlana E. Sedelnikova ◽

Thomas H. B. Schofield ◽

Patrick J. Baker

Keyword(s):

Serratia Marcescens ◽

Electron Density ◽

Crystallographic Analysis ◽

Gram Negative Bacteria ◽

Human Pathogen ◽

Gram Negative ◽

Anomalous Data ◽

Density Map ◽

Opportunistic Human Pathogen ◽

Electron Density Map

Tripartite α-pore-forming toxins are constructed of three proteins (A, B and C) and are found in many bacterial pathogens. While structures of the B and C components from Gram-negative bacteria have been described, the structure of the A component of a Gram-negative α-pore-forming toxin has so far proved elusive. SmhA, the A component from the opportunistic human pathogen Serratia marcescens, has been cloned, overexpressed and purified. Crystals were grown of selenomethionine-derivatized protein and anomalous data were collected. Phases were calculated and an initial electron-density map was produced.

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