A case report of Salmonella enterica serovar Corvallis from environmental isolates from Cambodia and clinical isolates in the UK

Salmonella enterica subspecies enterica serovar Corvallis (S. Corvallis) has been identified as a human pathogen and as a food contaminant. Diarrhoeal disease is a common diagnosis in tourists visiting Southeast Asia, often with unknown aetiology. However, numerous public health institutes have identified Salmonella as a common causative agent when consuming contaminated food and water. Genomic data from environmental isolates from a Cambodian informal market were uploaded to the National Center for Biotechnology Information (NCBI) platform, allowing the novel sequences to be compared to global whole-genome sequence archives. The comparison revealed that two human clinical isolates from England and four of the environmental isolates were closely related, with an average single nucleotide polymorphism (SNP) difference of 1 (0–3 SNPs). A maximum-likelihood tree based on core SNPs was generated comparing the 4 isolates recovered from a Cambodian informal market with 239 isolates of S. Corvallis received from routine surveillance of human salmonellosis in England and confirmed the close relationship. In addition, the environmental isolates clustered into a broader phylogenetic group within the S. Corvallis population containing 68 additional human isolates, of which 42 were from patients who reported recent international travel, almost exclusively to Southeast Asia. The environmental isolates of S. Corvallis isolated from an informal market in Cambodia are concerning for public health due to their genetic similarity to isolates (e.g. clinical isolates from the UK) with known human virulence and pathogenicity. This study emphasizes the benefits of global and public data sharing of pathogen genomes.

Molecular Genetic Analysis with Microsatellite-like Loci Reveals Specific Dairy-Associated and Environmental Populations of the Yeast Geotrichum candidum

Geotrichum candidum is an environmental yeast, also found as part of the cheese surface microbiota, where it is important in the ripening of many traditional cheeses, such as Camembert. We have previously developed a Multi Locus Sequence Typing (MLST) scheme, which differentiated five clades, of which one contained only environmental isolates, two were composed almost entirely of dairy isolates, and two others contained a mixture of dairy, environmental, and miscellaneous food isolates. In order to provide a simple method to uniquely type G. candidum strains, and in addition to permit investigation of the population structure at a fine level, we describe here a molecular analysis using a set of twelve highly discriminating microsatellite-like markers. The present study consolidates the previously suggested division between dairy and environmental strains, and in addition distinguishes a specifically European group of environmental strains. This analysis permitted the discrimination of 72 genotypes from the collection of 80 isolates, while retaining the underlying meaningful phylogenetic relation between groups of strains.

Phenotypical Comparison of Pseudomonas Aeruginosa Isolated from Human and Veterinary Samples; Impact of Host Adaptation on Infection Pathogenesis

Purpose: Determinants of virulence in Pseudomonas aeruginosa vary strongly depending on its habitat. In this study, we analyzed these alterations depending on the host organism in isolates cultured from canine ears and compared it to clinical extended-spectrum antibiotic-resistant Pseudomonas aeruginosa isolates (XDR), clinical antibiotic-sensitive (non-XDR) from humans and environmental isolates (EI) analyzed during our first study in 2017. Methods: A total of 22 veterinary isolates cultured from canine ears (VET) were examined for spontaneous biofilm formation, stress response in biofilm formation induced by meropenem, in vitro fitness, susceptibility to human serum and polymorphonuclear leukocytes and the genetically determined virulence factors toxA, exoS, exoT, exoU, exoY, nan1, cif, lasA and lasB.Results: We observed significantly elevated spontaneous biofilm formation and serum susceptibility in VET isolates compared to EI and non-XDR isolates as well as significantly decreased in vitro fitness compared to XDR isolates. The VET isolates resembled most the XDR subgroup of isolates previously cultured from blood. Within the environmental isolates, we observed an increase of spontaneous biofilm formation and exoU presence in isolates cultured from community water samples over hospital water samples to pool samples.Conclusions: Considering the distinct differences in some features of the examined VET isolates, a higher degree of phenotypical adaption can be assumed. Increased biofilm formation seems to be a common and characteristic event in isolates adapted to a specific habitat. Therefore amplification of potentially more virulent Pseudomonas aeruginosa strains in domestic animals may lead to elevated zoonotic risk for example for pet owners.

Clinical and Anatomopathological Evaluation of BALB/c Murine Models Infected with Isolates of Seven Pathogenic Sporothrix Species

Background: Sporotrichosis is a subcutaneous mycosis with worldwide distribution and caused by seven pathogenic species of Sporothrix genus: S. schenckii sensu stricto, S. brasiliensis, S. globosa and S. luriei (clinical clade), and the species S. mexicana, S. pallida and S. chilensis (environmental clade). Isolates of the same species of Sporothrix may have different pathogenicities; however, few isolates of this fungus have been studied. Thus, the aim of this work was to analyze the clinical and anatomopathological changes in immunocompetent and immunosuppressed BALB/c mice infected with clinical and environmental isolates of seven different species of Sporothrix, from both clades. One human clinical isolate of S. schenckii sensu stricto, S. brasiliensis, S. globosa, S. luriei, S. mexicana and S. chilensis species and one environmental isolate of S. pallida were inoculated subcutaneously in immunocompetent mice and the same isolates of S. brasiliensis and S.schenckii sensu stricto were inoculated in immunossupressed mice. Clinical manifestations as external lesions, apathy, and alopecia were observed. At 21, 35, and 49 days after fungal inoculation, four mice from each group were weighed, euthanized and necropsied for evaluation of splenic index, recovery of fungal cells, macroscopic and histopathological analysis of livers, lungs, kidneys, and hearts. The survival assessment was observed for 50 days following inoculation. Our results demonstrated that, clinical S. schenckii isolate, followed by clinical S. mexicana, and environmental S. pallida isolates, the last two, species grouped in the environmental clade, were capable of inducing greater anatomopathological changes in mice, which was reflected in the severity of the clinical signs of these animals. Thus, we reinforce the hypothesis that the pathogenicity of Sporothrix is not only related to the species of this fungus, but also shows variation between different isolates of the same species.

Comparative Genomics of Clinical and Environmental Isolates of Vibrio spp. of Colombia: Implications of Traits Associated with Virulence and Resistance

There is widespread concern about the increase in cases of human and animal infections caused by pathogenic Vibrio species due to the emergence of epidemic lineages. In Colombia, active surveillance by the National Institute of Health (INS) has confirmed the presence of Vibrio; however, in routine surveillance, these isolates are not genomically characterized. This study focused on the pangenome analysis of six Vibrio species: V. parahaemolyticus, V. vulnificus, V. alginolyticus, V. fluvialis, V. diabolicus and V. furnissii to determine the genetic architectures of potentially virulent and antimicrobial resistance traits. Isolates from environmental and clinical samples were genome sequenced, assembled and annotated. The most important species in public health were further characterized by multilocus sequence typing and phylogenomics. For V. parahaemolyticus, we found the virulent ST3 and ST120 genotypes. For V. vulnificus, we identified isolates belonging to lineages 1 and 2. Virulence gene homologues between species were found even in non-pathogenic species such as V. diabolicus. Annotations related to the mobilome, integrative mobile and conjugative elements and resistance genes were obtained from environmental and clinical isolates. This study contributes genomic information to the intensified surveillance program implemented by the INS to establish potential sources of vibriosis in Colombia.

Populating NCBI template for submissions using BioNumerics v7.6 v2

PURPOSE: to define the standard operating procedure for collecting isolate metadata using BioNumerics for submission of food/environmental isolates to NCBI. SCOPE: to provide a standardized procedure to collect isolate metadata using BioNumerics for submission of food/environmental isolates to NCBI. RESPONSIBILITIES- SOP Responsible Officials: Ruth Timme, Maria Balkey The GenomeTrakr Network Management will be responsible to monitor GenomeTrakr submissions processed through Bionumerics and ensure that all GT labs are familiar with the mandatory metadata fields required for submission of GenomeTrakr sequencing records to NCBI. V3: Dropdown menus from controlled vocabulary added to the ncbi_update submission sheet

Reoccurring Bovine Anthrax in Germany on the Same Pasture after 12 Years

The zoonotic disease anthrax caused by the endospore-forming bacterium Bacillus anthracis is very rare in Germany. In the state of Bavaria, the last case occurred in July of 2009 resulting in four dead cows. In August of 2021, the disease reemerged after heavy rains, killing one gestating cow. Notably, both outbreaks affected the same pasture, suggesting a close epidemiological connection. B. anthracis could be grown from blood culture and the presence of both virulence plasmids (pXO1 and pXO2) were confirmed by PCR. Also, recently developed diagnostic tools enabled rapid detection of B. anthracis cells and nucleic acids directly in clinical samples. The complete genome of the strain isolated from blood, designated BF-5, was DNA-sequenced and phylogenetically grouped within the B.Br.CNEVA clade that is typical for European B. anthracis strains. The genome was almost identical to BF-1, the isolate of 2009, separated only by three single nucleotide polymorphisms on the chromosome, one on plasmid pXO2 and three indel-regions. Further, B. anthracis DNA was detected by PCR from soil-samples taken from spots, where the cow had fallen onto the pasture. New tools based on phage receptor binding proteins enabled the microscopic detection and isolation of B. anthracis directly from soil-samples. These environmental isolates were genotyped and found to be SNP-identical to BF-1. Therefore, it seems that the BF-5 genotype is currently the prevalent one at the affected premises. The contaminated area was subsequently disinfected with formaldehyde.

Profile variation of bla genes among non-lactose fermenting Gram-negative bacilli between clinical and environmental isolates of Dr. Soetomo Hospital, Surabaya, Indonesia

Endraputra PN, Kuntaman K, Wasito EB, Shirakawa T, Raharjo D, Setyarini W. 2021. Profile variation of bla genes among non-lactose fermenting Gram negative bacilli between clinical and environmental isolates of Dr. Soetomo Hospital, Surabaya, Indonesia. Biodiversitas 22: 5047-5054. Carbapenem-resistant non-fermenter Gram-negative bacilli are notorious opportunistic pathogens in hospitalized patients and hospital environments. This study explored the carbapenemase gene among non-fermenter Gram-negative bacilli from hospital wastewater and clinical isolates in Dr. Soetomo Hospital, Surabaya, Indonesia. All samples were screened on MacConkey agar with meropenem 2 µg/ml and gene detected by Multiplex PCR. All samples were screened on MacConkey agar with meropenem 2 µg/ml and gene detected by Multiplex PCR. A total of 121 isolates consisted of 76 clinical (41 carbapenem-resistant Acinetobacter baumannii and 35 carbapenem-resistant Pseudomonas aeruginosa), 45 environmental isolates (6 carbapenem-resistant Pseudomonas aeruginosa and 32 carbapenem-resistant Pseudomonas spp.), and 7 screening samples (all CRPAs). Clinical isolates carbapenemase genes were identified, blaOXA-23-like 21 (28%), blaOXA-24-like 30 (39%), blaNDM-1 1 (1%), and blaIMP-1 6 (8%) while environmental isolates were blaOXA-23-like 5 (13%), blaOXA-24-like 4 (11%), blaOXA-48-like 2 (5%), blaNDM-1 13 (34%), blaVIM 12 (32%), and blaIMP-1 4 (11%). Rectal swab screening specimens presented blaOXA-23-like 3 (43%), blaOXA-24-like 3 (43%), and blaNDM-1 1 (14%). The carbapenemase gene pattern was different between clinical and environmental isolates. The blaOXA-23-like and blaOXA-24-like were most prevalent among in both clinical and wastewater, while blaVIM was mostly in wastewater. The presence of carbapenem-resistant non-fermenter Gram-negative bacilli carrying carbapenemase genes in hospital effluents indicated that the community river was seeded with an antimicrobial resistance gene.

Phylogenetic and antimicrobial drug resistance analysis of Vibrio cholerae O1 isolates from Ghana

We investigated the evolution, phylogeny and antimicrobial resistance of Vibrio cholerae O1 isolates (VCO1) from Ghana. Outbreak and environmental sources of VCO1 were characterized, whole-genome sequenced and compared to globally available seventh pandemic (7P) strains of V. cholerae at SNP resolution. Final analyses included 636 isolates. Novel Ghanaian isolates clustered into three distinct clades (clades 1, 2 and 3) in wave 3 of the 7P lineage. The closest relatives of our novel Ghanaian isolates were from Benin, Cameroon, Togo, Niger and Nigeria. All novel Ghanaian isolates were multi-drug resistant. Environmental isolates clustered into clade 2, despite being isolated years later, showing the possibility of persistence and re-emergence of older clades. A lag phase of several years from estimated introduction to reported cases suggests pathogen persistence in the absence of reported cholera cases. These results highlight the importance of deeper surveillance for understanding transmission routes between bordering countries and planning tailored vaccination campaigns in an effort to eradicate cholera.

Genetic Environment Surrounding bla OXA-55-like in Clinical Isolates of Shewanella algae Clade and Enhanced Expression of bla OXA-55-like in a Carbapenem-Resistant Isolate

Shewanella spp., which are known to carry chromosomally located bla OXA genes, have mainly been isolated from marine environments; however, they can also cause infections in humans. In this study, we compared the molecular characteristics of clinical isolates of Shewanella spp. with those originating from environmental sources. All 10 clinical isolates were genetically identified as members of the Shewanella algae clade ( S. algae , S. chilikensis , and S. carassii ); however, all but one of the 13 environmental isolates were identified as Shewanella species members outside the S. algae clade.