scholarly journals A Campylobacter integrative and conjugative element with a CRISPR-Cas9 system targeting competing plasmids: a history of plasmid warfare?

2021 ◽  
Vol 7 (11) ◽  
Author(s):  
Arnoud H. M. van Vliet ◽  
Oliver J. Charity ◽  
Mark Reuter
Keyword(s):  
Type Species ◽  
Campylobacter Coli ◽  
Bacterial Populations ◽  
Content Type ◽  
Microbial Genomes ◽  
Link Type ◽  
Cas9 Protein ◽  
History Of ◽  
Integrative Conjugative Elements ◽  
Integrative And Conjugative Element

Microbial genomes are highly adaptable, with mobile genetic elements (MGEs) such as integrative conjugative elements (ICEs) mediating the dissemination of new genetic information throughout bacterial populations. This is countered by defence mechanisms such as CRISPR-Cas systems, which limit invading MGEs by sequence-specific targeting. Here we report the distribution of the pVir, pTet and PCC42 plasmids and a new 70–129 kb ICE (CampyICE1) in the foodborne bacterial pathogens Campylobacter jejuni and Campylobacter coli . CampyICE1 contains a degenerated Type II-C CRISPR system consisting of a sole Cas9 protein, which is distinct from the previously described Cas9 proteins from C. jejuni and C. coli . CampyICE1 is conserved in structure and gene order, containing blocks of genes predicted to be involved in recombination, regulation and conjugation. CampyICE1 was detected in 134/5829 (2.3 %) C . jejuni genomes and 92/1347 (6.8 %) C . coli genomes. Similar ICEs were detected in a number of non-jejuni/coli Campylobacter species, although these lacked a CRISPR-Cas system. CampyICE1 carries three separate short CRISPR spacer arrays containing a combination of 108 unique spacers and 16 spacer-variant families. A total of 69 spacers and 10 spacer-variant families (63.7 %) were predicted to target Campylobacter plasmids. The presence of a functional CampyICE1 Cas9 protein and matching anti-plasmid spacers was associated with the absence of the pVir, pTet and pCC42 plasmids (188/214 genomes, 87.9 %), suggesting that the CampyICE1-encoded CRISPR-Cas has contributed to the exclusion of competing plasmids. In conclusion, the characteristics of the CRISPR-Cas9 system on CampyICE1 suggests a history of plasmid warfare in Campylobacter .

scholarly journals A Campylobacter integrative and conjugative element with a CRISPR-Cas9 system targeting competing plasmids: a history of plasmid warfare?

2021 ◽  
Author(s):  
Arnoud H. M. van Vliet ◽  
Oliver Charity ◽  
Mark Reuter
Keyword(s):  
Microbial Pathogens ◽  
Campylobacter Coli ◽  
Type Ii ◽  
Defence Mechanism ◽  
Bacterial Populations ◽  
Microbial Genomes ◽  
Cas9 Protein ◽  
History Of ◽  
Integrative Conjugative Elements ◽  
Integrative And Conjugative Element

Microbial genomes are highly adaptable, with mobile genetic elements (MGEs) such as integrative conjugative elements (ICE) mediating the dissemination of new genetic information throughout bacterial populations. This is countered by defence mechanism such as CRISPR-Cas systems, which limit invading MGEs by sequence-specific targeting. Here we report the distribution the pVir, pTet and PCC42 plasmids and a new 70-129 kb ICE (CampyICE1) in the foodborne microbial pathogens Campylobacter jejuni and Campylobacter coli. CampyICE1 contains a degenerated Type II-C CRISPR system consisting of a sole Cas9 protein, which is distinct from the previously described Cas9 proteins from C. jejuni and C. coli. CampyICE1 is conserved in structure and gene order, containing modules of genes predicted to be involved in recombination, regulation, and conjugation. CampyICE1 was detected in 134/5,829 (2.3%) C. jejuni genomes and 92/1,347 (6.8%) C. coli genomes. Similar ICE were detected in a number of non-jejuni/coli Campylobacter species, although these lacked a CRISPR-Cas system. CampyICE1 carries 3 separate short CRISPR spacer arrays containing a combination of 108 unique spacers and 16 spacer variant families, of which 70 spacers were predicted to target the Campylobacter plasmids pVir, pTet, and pCC42. A further nine spacers were predicted to target other Campylobacter plasmids (63.7%). The presence of a functional CampyICE1 Cas9 protein and matching anti-plasmid spacers was associated with the absence of these plasmids (188/214 genomes, 87.9%), implicating that the CampyICE1-encoded CRISPR-Cas has contributed to the exclusion of competing plasmids. In conclusion, the characteristics of the CRISPR-Cas9 system on CampyICE1 suggests a history of plasmid warfare in Campylobacter.

Conservation of Rhodococcus equi (Magnusson 1923) Goodfellow and Alderson 1977 and rejection of Corynebacterium hoagii (Morse 1912) Eberson 1918

2014 ◽  
Vol 64 (Pt_1) ◽  
pp. 311-312 ◽  
Author(s):  
George M. Garrity
Keyword(s):  
Type Species ◽  
Rhodococcus Equi ◽  
International Code ◽  
Content Type ◽  
Zoonotic Pathogen ◽  
Link Type ◽  
Code Of Nomenclature ◽  
History Of ◽  
International Code Of Nomenclature

A recent review of the nomenclatural history of Rhodococcus equi and its heterotypic synonyms reveals a situation in which the strict application of the Rules of the International Code of Nomenclature of Prokaryotes have resulted in the renaming of this known zoonotic pathogen, which may be reasonably viewed as a perilous name. This situation can be remedied only by the Judicial Commission rendering an opinion to conserve the name Rhodococcus equi and to reject its earlier heterotypic synonym, Corynebacterium hoagii .

scholarly journals The application of the CRISPR–Cas9 system in Pseudomonas syringae pv. actinidiae

2020 ◽  
Vol 69 (3) ◽  
pp. 478-486
Author(s):  
Joycelyn Ho ◽  
Min Zhao ◽  
Samuel Wojcik ◽  
George Taiaroa ◽  
Margi Butler ◽  
...  
Keyword(s):  
Pseudomonas Syringae ◽  
Type Species ◽  
Bacterial Pathogen ◽  
Specific Gene ◽  
Content Type ◽  
E Coli ◽  
Link Type ◽  
System A ◽  
Integrative Conjugative Element ◽  
Cas9 Protein

Introduction. Pseudomonas syringae pv. actinidiae (Psa) has emerged as a major bacterial pathogen of kiwifruit cultivation throughout the world. Aim. We aim to introduce a CRISPR–Cas9 system, a commonly used genome editing tool, into Psa. The protocols may also be useful in other Pseudomonas species. Methodology. Using standard molecular biology techniques, we modified plasmid pCas9, which carries the CRISPR–Cas9 sequences from Streptococcus pyogenes, for use in Psa. The final plasmid, pJH1, was produced in a series of steps and is maintained with selection in both Escherichia coli and Psa. Results. We have constructed plasmids carrying a CRISPR–Cas9 system based on that of S. pyogenes , which can be maintained, under selection, in Psa. We have shown that the gene targeting capacity of the CRISPR–Cas9 system is active and that the Cas9 protein is able to cleave the targeted sites. The Cas9 was directed to several different sites in the P. syringae genome. Using Cas9 we have generated Psa transformants that no longer carry the native plasmid present in Psa, and other transformants that lack the integrative, conjugative element, Pac_ICE1. Targeting of a specific gene, a chromosomal non-ribosomal peptide synthetase, led to gene knockouts with the transformants having deletions encompassing the target site. Conclusion. We have constructed shuttle plasmids carrying a CRISPR–Cas9 system that are maintained in both E. coli and P. syringae pv. actinidiae. We have used this gene editing system to eliminate features of the accessory genome (plasmids or ICEs) from Psa and to target a single chromosomal gene.

scholarly journals Insights into the acquisition of the pks island and production of colibactin in the Escherichia coli population

2021 ◽  
Vol 7 (5) ◽  
Author(s):  
Frédéric Auvray ◽  
Alexandre Perrat ◽  
Yoko Arimizu ◽  
Camille V. Chagneau ◽  
Nadège Bossuet-Greif ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Type Species ◽  
Whole Genome Sequence ◽  
Active Metabolites ◽  
Content Type ◽  
E Coli ◽  
Link Type ◽  
Sequence Types ◽  
Trna Locus ◽  
Integrative And Conjugative Element

The pks island codes for the enzymes necessary for synthesis of the genotoxin colibactin, which contributes to the virulence of Escherichia coli strains and is suspected of promoting colorectal cancer. From a collection of 785 human and bovine E. coli isolates, we identified 109 strains carrying a highly conserved pks island, mostly from phylogroup B2, but also from phylogroups A, B1 and D. Different scenarios of pks acquisition were deduced from whole genome sequence and phylogenetic analysis. In the main scenario, pks was introduced and stabilized into certain sequence types (STs) of the B2 phylogroup, such as ST73 and ST95, at the asnW tRNA locus located in the vicinity of the yersiniabactin-encoding High Pathogenicity Island (HPI). In a few B2 strains, pks inserted at the asnU or asnV tRNA loci close to the HPI and occasionally was located next to the remnant of an integrative and conjugative element. In a last scenario specific to B1/A strains, pks was acquired, independently of the HPI, at a non-tRNA locus. All the pks-positive strains except 18 produced colibactin. Sixteen strains contained mutations in clbB or clbD, or a fusion of clbJ and clbK and were no longer genotoxic but most of them still produced low amounts of potentially active metabolites associated with the pks island. One strain was fully metabolically inactive without pks alteration, but colibactin production was restored by overexpressing the ClbR regulator. In conclusion, the pks island is not restricted to human pathogenic B2 strains and is more widely distributed in the E. coli population, while preserving its functionality.

scholarly journals Bacterial gastroenteritis caused by the putative zoonotic pathogen Campylobacter lanienae: First reported case in Germany

2021 ◽  
Author(s):  
Juliane Fornefett ◽  
Anne Busch ◽  
Sandra Döpping ◽  
Helmut Hotzel ◽  
Dagmar Rimek
Keyword(s):  
Type Species ◽  
Campylobacter Coli ◽  
Pathogenic Potential ◽  
Content Type ◽  
Link Type ◽  
First Case ◽  
Strain Collection ◽  
Causative Agents ◽  
Bacterial Gastroenteritis ◽  
Abattoir Workers

Foodborne campylobacteriosis is the most common cause of human bacterial enteritis in Germany. Campylobacter jejuni and Campylobacter coli are the main causative agents for enteric disease, but a number of other species are involved, including rare ones. These rare Campylobacter spp. are emerging zoonotic pathogens in humans due to increasing international movement of supplies, livestock and people. Campylobacter lanienae was first isolated from healthy abattoir workers in Switzerland and at first its pathogenic potential for humans was considered to be low. Recently, the first case of Campylobacter lanienae -associated human enteritis was reported in Canada. Here, we describe a case of mild Campylobacter lanienae -associated enteritis with subsequent asymptomatic excretion in a butcher. The isolate is available at the TLV strain collection (no. TP00333/18). This first reported case of human Campylobacter lanienae campylobacteriosis in Germany demonstrates the agent’s likely zoonotic pathogenicity.

scholarly journals Wound infection caused by Neisseria zoodegmatis, a zoonotic pathogen: a case report

2021 ◽  
Author(s):  
John Merlino ◽  
Timothy Gray ◽  
Rohan Beresford ◽  
Sai Rupa Baskar ◽  
Thomas Gottlieb ◽  
...  
Keyword(s):  
Case Report ◽  
Type Species ◽  
Culture Media ◽  
Clinical History ◽  
Content Type ◽  
Zoonotic Pathogen ◽  
Link Type ◽  
Flight Mass Spectrometry ◽  
Clinical Laboratories ◽  
History Of

The isolation of Neisseria zoodegmatis from a 63-year-old female presenting to the emergency department following a cat bite injury to her right hand is described in this report. N. zoodegmatis , also known as Centers for Disease Control (CDC) group EF-4b, is considered to be a zoonotic pathogen, and is usually associated with dog or cat bites. Despite the potential of this organism to cause serious soft tissue infections, it can be overlooked in routine clinical laboratories due to its slow growth characteristics and when the history of animal bite is not provided to the laboratory. This case highlights the importance of appropriate clinical history provision to the microbiology laboratory to help provide important information about potential pathogens and allow microbiologists to optimize culture and identification methods. The introduction of tools such as matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) into clinical laboratories allows identification and the interpretation of results to be performed within a few minutes of isolation on proper culture media, as opposed to traditional methods, whose slowness may be problematic, as shown in this case report.

scholarly journals Intra-abdominal infection with Campylobacter curvus: case report and review of the literature

2021 ◽  
Vol 3 (4) ◽  
Author(s):  
Patrick C. K. Tam ◽  
Lex E. X. Leong ◽  
Maria Theodossi ◽  
David L. Gordon
Keyword(s):  
Type Species ◽  
Source Control ◽  
Alcoholic Pancreatitis ◽  
Oral Manifestations ◽  
Content Type ◽  
Oral Infections ◽  
Link Type ◽  
History Of ◽  
Minimal Data ◽  
Intra Abdominal Infection

Background. Campylobacter curvus is a Gram-negative bacteria associated with periodontal disease in humans. Cases of extra-oral manifestations of infection are rare with only six reported cases of extra-oral infection including this report that have been identified in the current literature. Molecular methods are generally used to identify C. curvus while optimal antibiotic choice and duration to treat extra-oral infections for this pathogen is unknown. Case presentation. A 63-year-old male with a background history of alcoholic pancreatitis presented with fever and malaise who was found to have radiological intra-abdominal collections. Drainage of these collections identified C. curvus via matrix-assisted laser desorption/ionisation time of flight (MALDI-TOF) mass spectrometry with high probability and identification further confirmed by whole-genome sequencing. Antibiotic susceptibility testing to erythromycin and ciprofloxacin of C. curvus was performed using E-test diffusion methods along with investigation for the presence of resistance genes. The patient was treated with intravenous piperacillin-tazobactam followed by ciprofloxacin for 4 weeks total with good clinical recovery. Conclusions. Extra-oral manifestations with the pathogen C. curvus are rare with few cases described in the literature. There is minimal data on susceptibility patterns, optimal antibiotic treatment and duration. Treatment of extraintestinal C. curvus infections in humans should encompass both adequate source control and antibiotic therapy.

scholarly journals Pseudomonas aeruginosa is capable of natural transformation in biofilms

Microbiology ◽  
2020 ◽  
Vol 166 (10) ◽  
pp. 995-1003 ◽  
Author(s):  
Laura M. Nolan ◽  
Lynne Turnbull ◽  
Marilyn Katrib ◽  
Sarah R. Osvath ◽  
Davide Losa ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Type Species ◽  
Natural Transformation ◽  
Bacterial Species ◽  
Extracellular Dna ◽  
Virulence Determinants ◽  
Bacterial Populations ◽  
Exogenous Dna ◽  
Content Type ◽  
Link Type

Natural transformation is a mechanism that enables competent bacteria to acquire naked, exogenous DNA from the environment. It is a key process that facilitates the dissemination of antibiotic resistance and virulence determinants throughout bacterial populations. Pseudomonas aeruginosa is an opportunistic Gram-negative pathogen that produces large quantities of extracellular DNA (eDNA) that is required for biofilm formation. P. aeruginosa has a remarkable level of genome plasticity and diversity that suggests a high degree of horizontal gene transfer and recombination but is thought to be incapable of natural transformation. Here we show that P. aeruginosa possesses homologues of all proteins known to be involved in natural transformation in other bacterial species. We found that P. aeruginosa in biofilms is competent for natural transformation of both genomic and plasmid DNA. Furthermore, we demonstrate that type-IV pili (T4P) facilitate but are not absolutely essential for natural transformation in P. aeruginosa .

scholarly journals Campylobacter novaezeelandiae sp. nov., isolated from birds and water in New Zealand

2020 ◽  
Vol 70 (6) ◽  
pp. 3775-3784 ◽  
Author(s):  
Samuel Bloomfield ◽  
David Wilkinson ◽  
Lynn Rogers ◽  
Patrick Biggs ◽  
Nigel French ◽  
...  
Keyword(s):  
New Zealand ◽  
Type Species ◽  
Genetic Relationships ◽  
Rrna Gene ◽  
Campylobacter Coli ◽  
Biochemical Tests ◽  
Whole Genome Analysis ◽  
Content Type ◽  
Link Type ◽  
Maldi Tof

Six isolates of Campylobacter with similar non-standard colonial morphologies were identified during studies isolating Campylobacter from bird faeces and rivers in New Zealand. Genomic (16S rRNA gene sequencing and whole genome analysis) and phenotypic (MALDI-TOF analysis and conventional biochemical tests) showed that the isolates form a monophyletic clade with genetic relationships to Campylobacter coli / Campylobacter jejuni and Campylobacter peloridis /Campylobacter amoricus. They may be distinguished from other Campylobacter by their MALDI-TOF spectral pattern, their florid α-haemolysis, their ability to grow anaerobically at 37 °C, and on 2 % NaCl nutrient agar, and their lack of hippuricase. This study shows that these isolates represent a novel species within the genus Campylobacter for which the name Campylobacter novaezeelandiae sp. nov. is proposed. The presence of C. novaezeelandiae in water may be a confounder for freshwater microbial risk assessment as they may not be pathogenic for humans. The type strain is B423bT (=NZRM 4741T=ATCC TSD-167T).

scholarly journals Infection with Clostridioides difficile ribotype 046 in a paediatric liver transplant patient

2021 ◽  
Vol 3 (10) ◽  
Author(s):  
Hildenia B. R. Nogueira ◽  
Cecília L. Costa ◽  
Conceição S. Martins ◽  
Maria Luana G. S. Morais ◽  
Carlos Quesada-Gómez ◽  
...  
Keyword(s):  
Molecular Characterization ◽  
Type Species ◽  
Transplant Patient ◽  
Antibiotic Use ◽  
Female Child ◽  
Content Type ◽  
Link Type ◽  
Clostridioides Difficile ◽  
History Of

Clostridioides difficile causes nosocomial diarrhoea associated with antibiotic use and immunodeficiency. Although the number of paediatric C. difficile infections (CDIs) has increased worldwide, there are few studies on the molecular characterization of strains causing CDIs among children. We report the clinical features and strain molecular characterization of a CDI in a female child with a history of liver transplantation at 7 months of age. This is the first report of the 046 ribotype causing paediatric diarrhoea.

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