The sixth global cholera pandemic lasted from 1899 to 1923. However, despite widespread fear of the disease and of its negative effects on troop morale, very few soldiers in the British Expeditionary Forces contracted cholera between 1914 and 1918. Here, we have revived and sequenced the genome of NCTC 30, a 102-year-old Vibrio cholerae isolate, which we believe is the oldest publicly available live V. cholerae strain in existence. NCTC 30 was isolated in 1916 from a British soldier convalescent in Egypt. We found that this strain does not encode cholera toxin, thought to be necessary to cause cholera, and is not part of V. cholerae lineages responsible for the pandemic disease. We also show that NCTC 30, which predates the introduction of penicillin-based antibiotics, harbours a functional β-lactamase antibiotic resistance gene. Our data corroborate and provide molecular explanations for previous phenotypic studies of NCTC 30 and provide a new high-quality genome sequence for historical, non-pandemic V. cholerae .
We take advantage of an historic collection of 133 Staphylococcus aureus strains accessioned between 1924 and 2016, whose genomes have been long-read sequenced as part of a major National Collection of Type Cultures (NCTC) initiative, to conduct a gene family-wide computational analysis of enterotoxin genes. We identify two novel Staphylococcal enterotoxin (pseudo)genes (sel29p and sel30), the former of which has not been observed in any contemporary strain to date. We provide further information on five additional enterotoxin genes or gene variants that have either recently entered the literature or for which the nomenclature or description is currently unclear (selz, sel26, sel27, sel28 and ses-2p). An examination of over 11,000 RefSeq genomes in search of wider support for these seven (pseudo)genes led to the identification of an additional three novel enterotoxin gene family members (sel31, sel32 and sel33) plus two new variants (seh-2p and ses-3p). We cast light on the genomic distribution of the enterotoxin genes, further defining their arrangement in gene clusters. Finally, we show that co-occurrence of enterotoxin genes is prevalent, with individual NCTC strains possessing as many as eighteen enterotoxin genes and pseudogenes, and that Clonal Complex membership rather than time of isolation is the key factor in determining enterotoxin load. Importance: Staphylococcus aureus strains pose a significant health risk to both human and animal populations. Key amongst this species’ virulence factors are the Staphylococcal enterotoxin gene family. Certain enterotoxin forms can induce a potentially life-threatening immune response, while others are implicated in less fatal though often severe conditions such as food poisoning. Genetic characterisation of Staphylococcal enterotoxin gene family members has steadily accumulated over recent decades, with over 20 genes now established in the literature. Despite the current wealth of knowledge on this important gene family, questions remain about the presence of additional enterotoxin genes and the genomic composition of family members. This study further expands knowledge of the Staphylococcal enterotoxins while shedding light on their evolution over the last century.
Kocuria rosea is a Gram-positive coccus found in the environment and within normal human skin microbiota, and more recently, it has been potentially implicated as an opportunistic pathogen. Here, we describe the genome sequences of five strains of K. rosea (NCTC2676, NCTC7514, NCTC7512, NCTC7528, and NCTC7511).
Variovorax durovernum nov. sp.was isolated from an infected, prosthetic endovascular graft explanted from a shepherd. Variovorax durovernum nov.sp. was isolated from blood agar, chocolate and MacConkey agar incubated at 37°C in an aerobic environment after 48h of incubation. Variovorax durovernum nov sp. is an aerobic, Gram-negative, motile, curved rod. Whole genome sequence (WGS) and targeted 16S sequencing was performed to retrieve nucleotide sequences representing the 16S rRNA gene of the isolate. The 16S gene were compared with all available Variovorax species.The 16S rRNA gene of Variovorax isolates differed by a minimum of 6 SNPs (99.6% identity) for V. paradoxus and 7 SNPs for V. boronicumulans (99.5%). The phylogeny of 16S gene placed Variovorax durovernum nov.sp. in a monophyletic cluster with closest relative being V. boronicumulans. The fatty acid profile consisted of 10:0 3OH (9.90%), Sum In Feature 3 (16:1 ω7c/16:1 ω6c - 16.36%), 16:0 (24.23%), 17:0 cyclo (13,35%), and Sum in Feature 8 (18:1 ω7c, 18:1 ω6c).In silico DNA-DNA hybridisation between the isolate and other Variovorax species was estimated at 32.1% for V. boronicumulans and 31.9% for V. paradoxus. Based on these results, Variovorax durovernum nov. sp. was designated as a novel species within the Variovorax genus, identified as the first case of human pathology caused by any Variovorax sp.
The National Collection of Type Cultures (NCTC) is a bacterial culture collection which is taxonomically and biologically diverse. NCTC holds approximately 6000 different strains from over 900 different species; among them strains originally isolated in the 19th century, strains for use as controls as stipulated EUCAST and ISO guidelines, type strains on which the description of bacterial species are based and other strains from a variety of backgrounds. The remit of NCTC is to provide authentic bacterial cultures of medical and veterinary interest to the scientific community, to support and enhance the reproducibility of scientific research and to improve global public health. To fulfil this remit, remain scientifically relevant and to preserve the legacy of contemporary medical bacteriology for future scientists, NCTC accessions strains of clinical significance: such as recently circulating and outbreak strains, diagnostic escape mutants and strains with novel antimicrobial resistance profiles. In 2018, 166 bacterial strains were accessioned into the NCTC and made available to the scientific community. These include NCTC 14052: a reference strain for emergent hyper-virulent K. pneumoniae, 4 type strains of newly described bacterial species, 82 strains accessioned from the Murray Collection of pre-antibiotic era Enterobacteriaceae and 8 strains with antimicrobial resistance mechanisms previously unrepresented in the collection, including NCTC 14208: a N. gonorrhoeae isolated from an instance of combined ceftriaxone and azithromycin treatment failure. Through literature review we have highlighted their value to the scientific community, both in their own right and in the context of bacterial strains already held by the NCTC.
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