Extensive genome analysis of Coxiella burnetii reveals limited evolution within genomic groups

This is the final version. Available on open access from BMC via the DOI in this recordAvailability of data and materials: Whole genome sequences were deposited in NCBI under BioProjects PRJNA430350 and PRJNA506366, as well as in the Sequence Read Archive as studies SRP130048 and SRP170036. Individual GenBank accession numbers for the WGS data are as follows: Q532 = PPFQ00000000.1 ; Q540 = PPFP00000000.1 ; Q545 = PPFO00000000.1 ; Q556 = PPFN00000000.1 ; Q559 = PPFM00000000.1 ; Cb_D1 = RQJU00000000.1; Cb_D2 = RQJT00000000.1 ; Cb_D8 = RQJS00000000.1 ; and Cb_D10 = RQJR00000000.1 .The authors declare that all other data supporting the findings of this study are available within the article and its supplementary information files.Background: Coxiella burnetii is a zoonotic pathogen that resides in wild and domesticated animals across the globe and causes a febrile illness, Q fever, in humans. An improved understanding of the genetic diversity of C. burnetii is essential for the development of diagnostics, vaccines and therapeutics, but genotyping data is lacking from many parts of the world. Sporadic outbreaks of Q fever have occurred in the United Kingdom, but the local genetic make-up of C. burnetii has not been studied in detail. Results: Here, we report whole genome data for nine C. burnetii sequences obtained in the UK. All four genomes of C. burnetii from cattle, as well as one sheep sample, belonged to Multi-spacer sequence type (MST) 20, whereas the goat samples were MST33 (three genomes) and MST32 (one genome), two genotypes that have not been described to be present in the UK to date. We established the phylogenetic relationship between the UK genomes and 67 publically available genomes based on single nucleotide polymorphisms (SNPs) in the core genome, which confirmed tight clustering of strains within genomic groups, but also indicated that sub-groups exist within those groups. Variation is mainly achieved through SNPs, many of which are non-synonymous, thereby confirming that evolution of C. burnetii is based on modification of existing genes. Finally, we discovered genomic-group specific genome content, which supports a model of clonal expansion of previously established genotypes, with large scale dissemination of some of these genotypes across continents being observed. Conclusions: The genetic make-up of C. burnetii in the UK is similar to the one in neighboring European countries. As a species, C. burnetii has been considered a clonal pathogen with low genetic diversity at the nucleotide level. Here, we present evidence for significant variation at the protein level between isolates of different genomic groups, which mainly affects secreted and membrane-associated proteins. Our results thereby increase our understanding of the global genetic diversity of C. burnetii and provide new insights into the evolution of this emerging zoonotic pathogen.Defence Science and Technology Laboratory (DSTL

Zoonoses under our noses.

This is the final version of the article. Available from Elsevier via the DOI in this record.One Health is an effective approach for the management of zoonotic disease in humans, animals and environments. Examples of the management of bacterial zoonoses in Europe and across the globe demonstrate that One Health approaches of international surveillance, information-sharing and appropriate intervention methods are required to successfully prevent and control disease outbreaks in both endemic and non-endemic regions. Additionally, a One Health approach enables effective preparation and response to bioterrorism threats.A.R.C. is supported by a BBSRC iCASE Studentship in partnership with the University of Exeter and the Defence Science and Technology Laboratory (Dstl) (Grand no.BB/M016404/1). S.R. is supported by the BBSRC grant number BB/N001591/1

Correlating genotyping data of coxiella burnetii with genomic groups

This is the final version. Available on open access from MDPI via the DOI in this recordData Availability Statement: All data are available in the Supplementary data file.Coxiella burnetii is a zoonotic pathogen that resides in wild and domesticated animals across the globe and causes a febrile illness, Q fever, in humans. Several distinct genetic lineages or genomic groups have been shown to exist, with evidence for different virulence potential of these lineages. Multispacer Sequence Typing (MST) and Multiple‐Locus Variable number tandem repeat Analysis (MLVA) are being used to genotype strains. However, it is unclear how these typing schemes correlate with each other or with the classification into different genomic groups. Here, we created extensive databases for published MLVA and MST genotypes of C. burnetii and analysed the associated metadata, revealing associations between animal host and human disease type. We established a new classification scheme that assigns both MST and MLVA genotypes to a genomic group and which revealed additional sub‐lineages in two genomic groups. Finally, we report a novel, rapid genomotyping method for assigning an isolate into a genomic group based on the Cox51 spacer sequence. We conclude that by pooling and streamlining existing datasets, associations between genotype and clinical outcome or host source were identified, which in combination with our novel genomotyping method, should enable an estimation of the disease potential of new C. burnetii isolates.Defence Science and Technology Laboratories (DSTL

Common Host Responses in Murine Aerosol Models of Infection Caused by Highly Virulent Gram-Negative Bacteria from the Genera Burkholderia, Francisella and Yersinia

This is the final version. Available on open access from MDPI via the DOI in this recordContent includes material subject to © Crown copyright (2019), Dstl. This material is licensed under the terms of the Open Government Licence except where otherwise stated. To view this licence, visit: http://www.nationalarchives.gov.uk/doc/open-government-licence/version/3 or write to the Information Policy Team, The National Archives, Kew, London TW9 4DU, or email: [email protected] virulent bacterial pathogens cause acute infections which are exceptionally difficult to treat with conventional antibiotic therapies alone. Understanding the chain of events that are triggered during an infection of a host has the potential to lead to new therapeutic strategies. For the first time, the transcriptomic responses within the lungs of Balb/C mice have been compared during an acute infection with the intracellular pathogens Burkholderia pseudomallei, Francisella tularensis and Yersinia pestis. Temporal changes were determined using RNAseq and a bioinformatics pipeline; expression of protein was also studied from the same sample. Collectively it was found that early transcriptomic responses within the infected host were associated with the (a) slowing down of critical cellular functions, (b) production of circulatory system components, (c) lung tissue integrity, and (d) intracellular regulatory processes. One common molecule was identified, Errfi1 (ErbB receptor feedback inhibitor 1); upregulated in response to all three pathogens and a potential novel marker of acute infection. Based upon the pro-inflammatory responses observed, we sought to synchronise each infection and report that 24 h p.i. of B. pseudomallei infection closely aligned with 48 h p.i. of infection with F. tularensis and Y. pestis. Post-transcriptional modulation of RANTES expression occurred across all pathogens, suggesting that these infections directly or indirectly modulate cell trafficking through chemokine expression/detection. Collectively, this unbiased NGS approach has provided an in-depth characterisation of the host transcriptome following infection with these highly virulent pathogens ultimately aiding in the development of host-directed therapies as adjuncts or alternatives to antibiotic treatment

Zoonoses under our noses.

One Health is an effective approach for the management of zoonotic disease in humans, animals and environments. Examples of the management of bacterial zoonoses in Europe and across the globe demonstrate that One Health approaches of international surveillance, information-sharing and appropriate intervention methods are required to successfully prevent and control disease outbreaks in both endemic and non-endemic regions. Additionally, a One Health approach enables effective preparation and response to bioterrorism threats

Passive strain distribution in the interosseous ligament of the forearm: Implications for injury reconstruction

Purpose: For severe forearm injuries such as an Essex-Lopresti fracture-dislocation, functional reconstruction necessitates repair of the interosseous ligament (IOL) to restore normal load sharing between the radius and ulna. Locating or tensioning such a reconstruction improperly can lead to abnormal load sharing and/or restriction of forearm rotation. The normal IOL strains should indicate the proper location of reconstruction grafts and the proper forearm rotation for tensioning the grafts. The objective of this study was to quantify the passive strain distribution of the IOL of the forearm with passive rotation of the forearm throughout the range of motion. Methods: The 3-dimensional motions of the radius with respect to the ulna were measured throughout forearm rotation in 10 cadaveric forearms by using an instrumented spatial linkage. From the bone motions and ligament insertion site geometry from dissection and computed tomographic scanning, insertion site motions were determined and used to calculate changes in ligament fiber lengths. Results: The measured strain distribution in the IOL was nonuniform and varied with forearm rotation. The overall magnitude of IOL strain was found to be greatest in supination and smallest in pronation. In supination the strains varied across fibers with strains being greatest in the distal fibers and lowest in the proximal fibers. Strains in neutral rotation were uniform across fibers. Although fibers were generally slack in pronation proximal fibers were less slack than distal fibers. Conclusions: The results of this study indicate that fiber strains in the IOL vary from proximal to distal and depend on forearm rotation. Our data suggest that to prevent restriction of forearm rotation all grafts should be tensioned in supination, where measured strains were generally highest. Our data also suggest that a 2-bundle IOL reconstruction may be necessary for proper load transfer between the radius and ulna in both supination and pronation. Copyright (C) 2004 by the American Society for Surgery of the Hand

Conscious surgery: influence of the environment on patient anxiety

Aims: i) To investigate anxiety arising from the experience of the clinical environment during surgery under local/ regional anaesthesia and, ii) to uncover the specific aspects patients find anxiety provoking and possibly dissuade them from opting for such anaesthesia. Background: Operating theatres have historical been designed for safe, efficient surgery on the unconscious patient and not primarily designed for the care of the ‘awake’ patient. However, with the rise in day surgery, the quantity of surgery performed under local/ regional anaesthesia is increasing. Method: As part of a larger study investigating anxiety within modern elective day surgery, adult patients undergoing surgery and local/ regional anaesthesia (n=214) were provided with a questionnaire on the day of surgery for return by mail 24 - 48 hours following surgery. Findings: The experience of being awake, possibly feeling surgeon, seeing body cut open or surgery being more painful were anxiety provoking aspects. Utilising factor analysis ‘intra-operative apprehension’, ‘anaesthetic information provision and ‘health control’ were identified as central features. Moreover, when employing multiple regression, apprehension associated with the intra-operative experience and anaesthetic information provision were significantly associated with an increase in the overall level of anxiety. Conclusions: Although the surrounding clinical environment has previously been a cause of apprehension, the sensations associated with the physical act of surgery on the conscious self appear also to have a considerable influence. Focusing care upon managing patient intra-operative experience and providing anaesthetic information in advance may help limit anxiety and expel the apparent misapprehensions associated with conscious surgery

The fate of Böhler's angle in conservatively-treated displaced intra-articular calcaneal fractures

Purpose: Although the predictive value of Böhler's angle on outcome remains subject of debate, the initial angle at the time of trauma still guides treatment. Changes in Böhler's angle during follow-up are frequently reported following surgical treatment of displaced intra-articular calcaneal fractures (DIACF). The aim of the present study was to determine the changes in Böhler's angle as a measure of secondary fracture displacement following conservative management of DIACF. Methods: Thirty-eight patients with a total of 44 displaced intra-articular calcaneal fractures treated conservatively with a minimum of two lateral radiographs during follow-up were analysed. Böhler's angle at different follow-up times was measured by three observers. The change in angle was compared with the angle at trauma, and influence of trauma mechanism and common calcaneal fracture classifications were determined. Results: The results showed a significant decline over time of the Böhler's angle in conservatively-treated patients of more than 11° on average at a mean follow-up of 29.2 weeks. This decrease was not related to gender, the initial angle, or the Essex-Lopresti or Sanders classification. A statistically significantly higher decrease was detected in high energetic trauma compared with low energetic trauma. Conclusion: The conservative treatment of displaced intra-articular calcaneal fractures is still a viable option, yet a significant secondary displacement in time should be taken into account, as reflected in a decrease of Böhler's angle of 11° up to one year following trauma

Current concepts in the treatment of intra-articular calcaneal fractures: results of a nationwide survey

The treatment of intra-articular calcaneal fractures is controversial and randomised clinical trials are scarce. Moreover, the socio-economic cost remains unclear. The aim of this study was to estimate the incidence, treatment preferences and socio-economic cost of this complex fracture in the Netherlands. This data may aid in planning future clinical trials and support education. The method of study was of a cross-sectional survey design. A written survey was sent to one representative of both the traumatology and the orthopaedic staff in each hospital in the Netherlands. Data on incidence, treatment modalities, complications and follow-up strategies were recorded. The socio-economic cost was calculated. The average response rate was 70%. Fracture classifications, mostly by Sanders and Essex-Lopresti, were applied by 29%. Annually, 920 intra-articular calcaneal fractures (0.4% incidence rate) were treated, mainly with ORIF (46%), conservative (39%) and percutaneous (10%) treatment. The average non-weight-bearing mobilisation was 9 weeks (SD 2 weeks). An outcome score, mainly AOFAS, was documented by 7%. A secondary arthrodesis was performed in 21% of patients. The socio-economic cost was estimated to be €21.5–30.7 million. Dutch intra-articular calcaneal fracture incidence is at least 0.4% of all fractures presenting to hospitals. Better insight into treatment modalities currently employed and costs in the Netherlands was obtained