Computed tomography (CT) and magnetic resonance imaging (MRI) examinations of deceased individuals are increasingly being utilized in the field of forensic pathology. However, there are differences ...in the interpretation of post-mortem and clinical imaging. Radiologists with only occasional experience in post-mortem imaging are at risk of misinterpreting the findings if they rely solely on clinical experience. Radiological specialists working in a co-operative environment with pathologists are pivotal in the understanding of post-mortem CT and MRI, and its appropriate integration into the autopsy. This has spawned a novel subspecialty called post-mortem radiology or necro-radiology (radiology of the deceased). In the future it is likely that whole-body CT will be incorporated into the routine forensic autopsy due its ability to accurately detect and localise abnormalities commonly seen in forensic practice, such as haematoma, abnormal gas collections, fractures, and metallic foreign bodies. In the next 5–10 years most forensic institutes will seek regular access to such CT facilities or install machines into their own mortuaries. MRI is technically more problematic in the deceased but the improved tissue contrast over CT means that it is also very useful for investigation of pathology in the cranial, thoracic, and abdominal cavities, as well as the detection of haematoma in soft tissue. In order for radiologists to be an integral part of this important development in forensic investigation, radiological organizations must recognize the subspecialty of post-mortem radiology and provide a forum for radiologists to advance scientific knowledge in the field.
Background
The global accumulation of Escherichia coli with CTX-M extended-spectrum β-lactamases partly reflects the dissemination of clonal lineages, notably ST131 and ST405. More recently, E. coli ...have emerged that produce NDM carbapenemase. We sought to determine the clonal diversity of E. coli with this enzyme from English hospitals, and to compare them with isolates from Pakistan and India.
Methods
The 18 NDM-positive E. coli were from hospitals in England (n = 10), Pakistan (n = 7) and India (n = 1). Isolates were compared by phylogenetic grouping, multilocus sequence typing and PFGE of XbaI-digested DNA. Isolates were screened by PCR for acquired AmpC genes, bla
CTX-M, and the 16S rRNA methylase genes armA and rmtC.
Results
Most of the isolates belonged to phylogenetic groups B1 (n = 9) or D (n = 7); two were group A and none was group B2. Nine isolates from England and Pakistan belonged to the B1 lineage ST101, with seven of these clustering at >77% similarity by PFGE. Other lineages included ST405 (n = 3, group D), ST648 (n = 3, group D), the ST23 complex (one each of ST90 and ST410, both group A) and ST156 (n = 1, group D). Sixteen of 18 isolates had a group 1 CTX-M gene, 13 had a CIT-type acquired AmpC, and 16 had either or both of armA and rmtC.
Conclusions
The E. coli isolates producing NDM-1 carbapenemase belonged to six sequence types and included diverse clonal lineages. Nevertheless, isolates of B1-ST101 accounted for half the collection, and included isolates from both England and Pakistan. None of the isolates belonged to ST131 or to phylogroup B2.
We determined the prevalence and types of extended-spectrum β-lactamase (ESBL)-producing and carbapenem-resistant Escherichia coli in raw retail beef, chicken, pork, fruit and vegetables in five UK ...regions in 2013–14. Raw meat (n=397), and fruit and vegetable samples (n=400) were purchased from retail stores in London, East Anglia, North West England, Scotland and Wales. Samples were tested for the presence of ESBL-producing E. coli by plating enriched samples on CHROMagar CTX and CHROMagar ESBL, for AmpC-type E. coli by plating on “CHROMagar FOX” (CHROMagar ECC+16mg/L cefoxitin), and for carbapenem-resistant E. coli by plating on CHROMagar KPC. Additionally, pre-enrichment counts were performed on the above agars, and on CHROMagar ECC. Isolates of interest were characterised by MALDI-ToF to confirm identification, by PCR for blaCIT,blaCTX-M,blaOXA, blaSHV and blaTEM genes; ESBL or blaCIT genes were sequenced. Only 1.9% and 2.5% of beef and pork samples, respectively were positive for ESBL-producing E. coli after enrichment compared with 65.4% of chicken samples. 85.6% positive samples from chicken meat carried blaCTX-M-1; blaCTX-M-15 was not detected. None of the fruits or vegetables yielded ESBL-producing E. coli and none of the meat, fruit or vegetable samples yielded carbapenem-resistant E. coli. Retail chicken was more frequently a source of ESBL-producing E. coli than were beef, pork, fruit or vegetables. None of the foodstuffs yielded E. coli with CTX-M-15 ESBL, which dominates in human clinical isolates in the UK, and none yielded carbapenem-resistant E. coli.
•Retail raw meat, fruit and vegetables were tested for antibiotic-resistant E. coli.•Samples were purchased in the UK in 2013 to 2014.•About 2% of beef and pork and 65% of chicken were positive for ESBL-producing E. coli.•None of the fruit and vegetable samples was positive for ESBL-producing E. coli.•Carbapenem-resistant E. coli were not isolated from any of the samples tested.
Objectives The aim of this study was to investigate relatedness and molecular mechanism(s) of ertapenem resistance in clinical isolates of Klebsiella spp. (n = 28) and Enterobacter spp. (n = 27) ...referred from multiple hospitals to the UK national reference laboratory. Methods Investigations included genotyping by PFGE, resistance gene analysis by PCR and antimicrobial susceptibility testing with and without inhibitors of efflux and β-lactamase activity. Outer membrane proteins (OMPs) were profiled by SDS–PAGE; porin genes were sequenced and their expression was examined by RT–PCR. The contribution of porin deficiency to resistance was investigated by restoring functional porin genes on plasmids. Results PFGE showed significant clonal diversity among ertapenem-resistant isolates, with only small clusters identified. SHV- and CTX-M-type extended-spectrum β-lactamases were identified in the Klebsiella spp. isolates, whereas AmpC overexpression or KPC carbapenemase was detected in the Enterobacter cloacae isolates. SDS–PAGE showed that Klebsiella pneumoniae and Enterobacter aerogenes with high-level ertapenem resistance (MICs ≥ 16 mg/L) consistently lacked both of the two major non-specific porins, whereas variable patterns of OmpC and OmpF were seen in E. cloacae with lower-level ertapenem resistance. Various point mutations or insertion sequences were identified as disrupting the porin-coding sequences, as well as mutations in the promoter region. Functional restoration of OmpK35 or OmpK36 in Klebsiella and OmpC or OmpF in Enterobacter spp. isolates significantly decreased the MICs of all carbapenems, but particularly of ertapenem. We found no evidence of efflux contributing to resistance. Conclusions Ertapenem resistance was exclusively due to combinations of β-lactamases with impermeability caused by loss of OMPs. Efflux was not implicated and there was no national spread of resistant clones.
The accurate performance of antimicrobial susceptibility testing of bacteria from animal sources and the correct presentation of the results is a complex matter. A review of the published literature ...revealed a number of recurring errors with regard to methodology, quality control, appropriate interpretive criteria, and calculation of MIC50 and MIC90 values. Although more subjective, there is also no consensus regarding the definition of multiresistance. This Editorial is intended to provide guidance to authors on how to avoid these frequently detected shortcomings.
Abstract CT scanning of the deceased is an established technique performed on all individuals admitted to VIFM over the last 5 years. It is used primarily to assist pathologists in determining cause ...and manner of death but is also invaluable for identification of unknown deceased individuals where traditional methods are not possible. Based on this experience, CT scanning was incorporated into phase 2 of the Institute's DVI process for the 2009 Victorian bushfires. All deceased individuals and fragmented remains admitted to the mortuary were CT scanned in their body bags using established protocols. Images were reviewed by 2 teams of 2 radiologists experienced in forensic imaging and the findings transcribed onto a data sheet constructed specifically for the DVI exercise. The contents of 255 body bags were examined in the 28 days following the fires. 164 missing persons were included in the DVI process with 163 deceased individuals eventually identified. CT contributed to this identification in 161 persons. In 2 cases, radiologists were unable to recognize commingled remains. CT was utilized in the initial triage of each bag's contents. If radiological evaluation determined that bodies were incomplete then this information was provided to search teams who revisited the scenes of death. CT was helpful in differentiation of human from non-human remains in 8 bags, recognition of human/animal commingling in 10 bags and human commingling in 6 bags. In 61% of cases gender was able to be determined on CT using a novel technique of genitalia detection and in all but 2 cases this was correct. Age range was able to be determined on CT in 94% with an accuracy of 76%. Specific identification features detected on CT included the presence of disease (14 disease entities in 13 cases), medical devices (26 devices in 19 cases) and 274 everyday metallic items associated with the remains of 135 individuals. CT scanning provided useful information prior to autopsy by flagging likely findings including the presence of non-human remains, at the time of autopsy by assisting in the localization of identifying features in heavily disfigured bodies, and after autopsy by retrospective review of images for clarification of issues that arose at the time of pathologist case review. In view of the success of CT scanning in this mass disaster, DVI administrators should explore the incorporation of CT services into their disaster plans.
The development of antibiotic resistance by bacteria is an evolutionary inevitability, a convincing demonstration of their ability to adapt to adverse environmental conditions. Since the emergence of ...penicillinase-producing Staphylococcus aureus in the 1940s, staphylococci, enterococci and streptococci have proved themselves adept at developing or acquiring mechanisms that confer resistance to all clinically available antibacterial classes. The increasing problems of methicillin-resistant S. aureus and coagulase-negative staphylococci (MRSA and MRCoNS), glycopeptide-resistant enterococci and penicillin-resistant pneumococci in the 1980s, and recognition of glycopeptide-intermediate S. aureus in the 1990s and, most recently, of fully vancomycin-resistant isolates of S. aureus have emphasised our need for new anti-Gram-positive agents. Antibiotic resistance is one of the major public health concerns for the beginning of the 21st century. The pharmaceutical industry has responded with the development of oxazolidinones, lipopeptides, injectable streptogramins, ketolides, glycylcyclines, second-generation glycopeptides and novel fluoroquinolones. However, clinical use of these novel agents will cause new selective pressures and will continue to drive the development of resistance. This review describes the various antibiotic resistance mechanisms identified in isolates of staphylococci, enterococci and streptococci, including mechanisms of resistance to recently introduced anti-Gram-positive agents.
the emergence of carbapenemases in Enterobacteriaceae is driving a search for therapeutic alternatives. We tested ACHN-490, a sisomicin derivative that evades all plasmid-mediated ...aminoglycoside-modifying enzymes, against 82 carbapenem-resistant Enterobacteriaceae isolates. Comparators included internationally and locally available aminoglycosides. Methods The isolates variously had KPC (n = 12), SME-1 (n = 1), IMP (n = 13), VIM (n = 5), NDM (n = 17) or OXA-48 (n = 19) carbapenemases, or had combinations of impermeability with AmpC (n = 5) or extended-spectrum β-lactamases (n = 10). They included 53 Klebsiella spp., 19 Enterobacter spp., 6 Escherichia coli and 4 others; most were multiresistant. Genes were identified by PCR and sequencing; MICs were measured by CLSI agar dilution.
ACHN-490 was active at ≤ 2 mg/L against all 65 isolates with carbapenem resistance mechanisms other than NDM enzyme, mostly with MICs of 0.12-0.5 mg/L; isepamicin was active against 63/65 at ≤ 8 mg/L. In contrast, 35% were resistant to gentamicin at 4 mg/L, 61% to tobramycin at 4 mg/L and 20% to amikacin at 16 mg/L. However, 16 of the 17 isolates with NDM-1 enzyme were resistant to ACHN-490, with MICs ≥ 64 mg/L, and these were cross-resistant to all other human-use aminoglycosides tested. Their behaviour was associated with ArmA and RmtC 16S rRNA methylases. Apramycin (a veterinary aminoglycoside) retained its full activity, with MICs of 4-8 mg/L versus strains with armA or rmtC, though resistance was seen in one Klebsiella pneumoniae with AAC(3)-IV (MIC ≥ 256 mg/L).
ACHN-490 has potent activity versus carbapenem-resistant isolates, except those also harbouring 16S rRNA methylases; isepamicin is also widely active, though less potent than ACHN-490. Evasion of 16S rRNA methylases by apramycin is noteworthy and may provide a starting point for future aminoglycoside development.
As an emerging technology, Rapid DNA has demonstrated its utility for law enforcement in the provision of DNA profiling data at the point of arrest, often not requiring analyst review of the profiles ...generated. Recently, efforts have centred on the evaluation of Rapid DNA (without analyst review) and modified Rapid DNA (requiring review by a trained analyst) for application to crime scene samples. In a broader forensic context, however, another application for Rapid DNA is its use to process post-mortem samples to assist with the identification of deceased persons; and while gaps in our knowledge remain as to how Rapid DNA instruments perform with these sample types (often compromised with regards to their yield and quality of DNA), they have been successfully deployed in the field to assist in the identification of disaster victims (as exemplified during the 2018 Californian wildfire). This review aims to provide the current research landscape for the forensic application of Rapid DNA as an emerging technology from a Disaster Victim Identification perspective.
•Rapid DNA offers the possibility to take DNA profiling out of the laboratory.•Commercial instruments are reliable for processing buccal samples.•Rapid DNA could assist in DVI by reducing profiling time and simplifying logistics.•Benefit in DVI will depend on operational and jurisdictional circumstances.
Enterobacteriaceae producing carbapenemases, such as KPC or metallo-β-lactamases (MBLs), have emerged on several continents. Phenotypic tests are urgently needed for their rapid and accurate ...detection. A novel carbapenemase detection test, comprising a meropenem disk, and meropenem disks supplemented with 730 μg of EDTA, 1000 μg of dipicolinic acid (DPA), 600 μg of aminophenylboronic acid (APBA), or 750 μg of cloxacillin, was evaluated against Klebsiella pneumoniae isolates with KPC (n = 34), VIM (n = 21), IMP (n = 4) or OXA-48 (n = 9) carbapenemases, and carbapenem-resistant Enterobacteriaceae with porin loss in combination with an extended-spectrum β-lactamase (ESBL) (n = 9) or AmpC hyperproduction (n = 5). Commercially available diagnostics tablets from Rosco containing meropenem and the same inhibitors as described above (except EDTA) were also evaluated. An increased meropenem inhibition zone was sought in the presence of each added β-lactamase inhibitor. APBA had excellent sensitivity for detecting K. pneumoniae with KPC enzymes. Isolates with combined AmpC hyperproduction and porin loss were also positive in the APBA test but, unlike KPC producers, showed cloxacillin synergy. Both DPA and EDTA had excellent sensitivity for detection of MBL-producing K. pneumoniae. However, EDTA showed poor specificity, with positive results noted for 1/9 ESBL-producing isolates, for 4/34 KPC-producing isolates, and for 4/9 OXA-48-producing isolates, whereas all of these were negative when DPA was used. The in-house test distinguished accurately between several different mechanisms mediating reduced susceptibility to carbapenems in Enterobacteriaceae. The commercial combination tablets from Rosco performed similarly to the in-house test, with the exception of one false-positive MBL result and one false-positive KPC result among the OXA-48 producers.
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