UK Isolates Research Articles

Gene × environment tests discriminate the new EU2 evolutionary lineage of Phytophthora ramorum and indicate that it is adaptively different

SummaryA new evolutionary lineage of the destructive introduced tree pathogen Phytophthora ramorum, EU2 lineage, was recently discovered attacking larch and other hosts in Northern Ireland and south west Scotland, UK. Sixteen ‘medium × agar concentration × incubation temperature’ stress environments were tested to find a rapid and repeatable method to discriminate the known EU2 lineage from the EU1, NA1 and NA2 lineages in culture, in particular from the EU1 already prevalent across the UK; and to investigate whether EU2 might be adaptively different. At 25°C on both Carrot agar and V8 juice agar, the mean radial growth rates of all four lineages were significantly different, with NA2 > EU2 > EU1 > NA1. At this temperature, EU2 colonies were not only phenotypically distinct from EU1 and all other lineages but on average grew three times as fast as EU1. This indicates that EU2 is adaptively different from EU1. Twelve days growth in the environment ‘V8A/2% agar/25°C gave excellent discrimination of all four lineages in three repeat experiments, including clear discrimination of EU2 from EU1. Each lineage exhibited a distinctive colony pattern. The utility of this test environment was examined further by screening fresh UK isolates of unknown lineage from new larch outbreak sites alongside standard isolates. The lineage assignments predicted were corroborated by gene sequencing and RFLP profiling. These results also revealed that the EU2 lineage was present at several new larch sites in south west Scotland, whereas isolates from geographically adjacent areas such as the Isle of Mull, north west Scotland, the Isle of Man and north west England were all of EU1 lineage.

Genome analyses of the wheat yellow (stripe) rust pathogen Puccinia striiformis f. sp. triticireveal polymorphic and haustorial expressed secreted proteins as candidate effectors

BackgroundWheat yellow (stripe) rust caused by Puccinia striiformis f. sp. tritici (PST) is one of the most devastating diseases of wheat worldwide. To design effective breeding strategies that maximize the potential for durable disease resistance it is important to understand the molecular basis of PST pathogenicity. In particular, the characterisation of the structure, function and evolutionary dynamics of secreted effector proteins that are detected by host immune receptors can help guide and prioritize breeding efforts. However, to date, our knowledge of the effector repertoire of cereal rust pathogens is limited.ResultsWe re-sequenced genomes of four PST isolates from the US and UK to identify effector candidates and relate them to their distinct virulence profiles. First, we assessed SNP frequencies between all isolates, with heterokaryotic SNPs being over tenfold more frequent (5.29 ± 2.23 SNPs/kb) than homokaryotic SNPs (0.41 ± 0.28 SNPs/kb). Next, we implemented a bioinformatics pipeline to integrate genomics, transcriptomics, and effector-focused annotations to identify and classify effector candidates in PST. RNAseq analysis highlighted transcripts encoding secreted proteins that were significantly enriched in haustoria compared to infected tissue. The expression of 22 candidate effector genes was characterised using qRT-PCR, revealing distinct temporal expression patterns during infection in wheat. Lastly, we identified proteins that displayed non-synonymous substitutions specifically between the two UK isolates PST-87/7 and PST-08/21, which differ in virulence to two wheat varieties. By focusing on polymorphic variants enriched in haustoria, we identified five polymorphic effector candidates between PST-87/7 and PST-08/21 among 2,999 secreted proteins. These allelic variants are now a priority for functional validation as virulence/avirulence effectors in the corresponding wheat varieties.ConclusionsIntegration of genomics, transcriptomics, and effector-directed annotation of PST isolates has enabled us to move beyond the single isolate-directed catalogues of effector proteins and develop a framework for mining effector proteins in closely related isolates and relate these back to their defined virulence profiles. This should ultimately lead to more comprehensive understanding of the PST pathogenesis system, an important first step towards developing more effective surveillance and management strategies for one of the most devastating pathogens of wheat.

Association of the novel aminoglycoside resistance determinant RmtF with NDM carbapenemase in Enterobacteriaceae isolated in India and the UK

16S rRNA methyltransferases are an emerging mechanism conferring high-level resistance to clinically relevant aminoglycosides and have been associated with important mechanisms such as NDM-1. We sought genes encoding these enzymes in isolates highly resistant (MIC >200 mg/L) to gentamicin and amikacin from an Indian hospital and we additionally screened for the novel RmtF enzyme in 132 UK isolates containing NDM. All highly aminoglycoside-resistant isolates were screened for armA and rmtA-E by PCR, with cloning experiments performed for isolates negative for these genes. Matrix-assisted laser desorption/ionization (MALDI) mass spectrometry was used to determine the methylation target of the novel RmtF methyltransferase. RmtF-bearing strains were characterized further, including susceptibility testing, PFGE, electroporation, PCR-based replicon typing and multilocus sequence typing of rmtF-bearing plasmids. High-level aminoglycoside resistance was detected in 140/1000 (14%) consecutive isolates of Enterobacteriaceae from India. ArmA, RmtB and RmtC were identified among 46%, 20% and 27% of these isolates, respectively. The novel rmtF gene was detected in 34 aminoglycoside-resistant isolates (overall prevalence 3.4%), most (59%) of which also possessed a bla(NDM) gene; rmtF was detected in 6 NDM producers from the UK. It was found on different plasmid backbones. Four and two isolates showed resistance to tigecycline and colistin, respectively. RmtF was often found in association with NDM in members of the Enterobacteriaceae and on diverse plasmids. It is of clinical concern that the RmtF- and NDM-positive strains identified here show additional resistance to tigecycline and colistin, current drugs of last resort for the treatment of serious bacterial infections.

First Report of Potato spindle tuber viroid Naturally Infecting Greenhouse Tomatoes in North Carolina.

In spring 2012, a severe disease was observed on a limited number of tomato plants (Solanum lycopersicum L.) in a research greenhouse facility in western North Carolina. The first symptoms noted were downward curling of the terminal leaves accompanied by a rough puckered darker green texture. This was followed in time by greater distortion of the leaves with pale green on leaf margins. Older leaves with symptoms developed necrosis, with necrotic spots and streaks appearing on a few fruits. On some of these affected fruits, stems, peduncles, pedicels, and sepals also showed symptoms. Infected plants were badly stunted, and fruits in the upper parts of plants displaying severe symptoms remained very small. In just a few months, the disease spread to other tomato plants inside the greenhouse. A survey in May 2012 showed a disease incidence of 18% (156 symptomatic plants out of a total of 864) in this greenhouse. Initial screenings for possible viruses using ELISA (Agdia, Elkhart, IN), as well as a reverse transcription (RT)-PCR panel of 15 common tomato viruses in our laboratory were negative. Because of the symptoms and negative results for viruses, a viroid infection was suspected. Total plant RNA was prepared using TRIzol reagent (Invitrogen, Carlsbad, CA) from leaf tissues of eight diseased plants and one seed sample. Using real-time RT-PCR developed against Potato spindle tuber viroid (PSTVd) and some related pospiviroids (1), positive signals were observed with a mean Ct = 13.24 for leaf tissues and Ct = 19.91 for the seed sample. To obtain a full viroid genome, RT-PCR using two different sets of primers, one specific for PSTVd (PSTVd-F and PSTVd-R) (2), and a universal primer set for pospiviroids (MTTVd-F and MTTVd-R) (3) was performed. RT-PCR generated amplicons with expected size of ~360 bp from all eight leaf and one seed samples, but not from a healthy control. PCR products were cloned using the TOPO TA cloning kit (Invitrogen, Carlsbad, CA). A total of 22 full genomic sequences were obtained. A multi-sequence alignment generated a consensus sequence of 360 nt, designated as NC12-01 (GenBank Accession No. JX280944). BLASTn search in the NCBI database revealed the highest sequence identity of 96.9% to Australian (AY962324) and UK (AJ583449) isolates of PSTVd and 95.9% identity to the tomato isolate of PSTVd-CA1 (HM753555). Similar disease symptoms were observed on two 'Rutgers' tomato plants 2 weeks post mechanical inoculation and the presence of PSTVd was confirmed by real-time RT-PCR (1). A mock-inoculated plant did not show any symptoms. In the U.S., natural infection of PSTVd on tomato was first identified in California in 2010 (3). To our knowledge, this is the first report of a natural occurrence of PSTVd on tomato in the eastern U.S. The diseased plants were contained, properly disposed of, and eradicated in this location. The broader geographic distribution of PSTVd on tomato in the U.S., and the potential latent infection in potato and a number of ornamentals (4), emphasizes the need for better plant and seed health tests for viroids on these plants.

Insights into a Multidrug Resistant Escherichia coli Pathogen of the Globally Disseminated ST131 Lineage: Genome Analysis and Virulence Mechanisms

Escherichia coli strains causing urinary tract infection (UTI) are increasingly recognized as belonging to specific clones. E. coli clone O25b:H4-ST131 has recently emerged globally as a leading multi-drug resistant pathogen causing urinary tract and bloodstream infections in hospitals and the community. While most molecular studies to date examine the mechanisms conferring multi-drug resistance in E. coli ST131, relatively little is known about their virulence potential. Here we examined E. coli ST131 clinical isolates from two geographically diverse collections, one representing the major pathogenic lineages causing UTI across the United Kingdom and a second representing UTI isolates from patients presenting at two large hospitals in Australia. We determined a draft genome sequence for one representative isolate, E. coli EC958, which produced CTX-M-15 extended-spectrum β-lactamase, CMY-23 type AmpC cephalosporinase and was resistant to ciprofloxacin. Comparative genome analysis indicated that EC958 encodes virulence genes commonly associated with uropathogenic E. coli (UPEC). The genome sequence of EC958 revealed a transposon insertion in the fimB gene encoding the activator of type 1 fimbriae, an important UPEC bladder colonization factor. We identified the same fimB transposon insertion in 59% of the ST131 UK isolates, as well as 71% of ST131 isolates from Australia, suggesting this mutation is common among E. coli ST131 strains. Insertional inactivation of fimB resulted in a phenotype resembling a slower off-to-on switching for type 1 fimbriae. Type 1 fimbriae expression could still be induced in fimB-null isolates; this correlated strongly with adherence to and invasion of human bladder cells and bladder colonisation in a mouse UTI model. We conclude that E. coli ST131 is a geographically widespread, antibiotic resistant clone that has the capacity to produce numerous virulence factors associated with UTI.

Genetics of resistance to downy mildew in Brassica oleracea and breeding towards durable disease control for UK vegetable production

Downy mildew resistance was previously identified from screening a Brassica oleracea collection against two standard UK isolates of Hyaloperonospora parasitica. Sources of resistance were chosen from this material and developed further in this study by generating doubled haploid (DH) and inbred lines. Seedlings from the new lines were tested for resistance to a larger collection of H. parasitica isolates collected in 2001–2002 and 2007–2008 from the main broccoli and cauliflower production regions of the UK. Three lines (derived from borecole or summer cabbage) were broadly resistant to the pathogen isolates. Three of the remaining lines exhibited strong isolate‐specific resistance; several examples of weak or basal level of resistance to some isolates were observed. A new H. parasitica variant collected in 2008 was virulent in the broadly resistant lines, but was avirulent in a line with narrow specificity of resistance. The F2 and BC1 seedlings derived from outcrossing each of the three broadly resistant lines to susceptible broccoli and cauliflower lines segregated in a manner indicating that the resistance was controlled by a single dominant gene. No susceptibility was observed amongst F2 seedlings derived from intercrossing the three resistant lines, indicating that they all share the same or closely linked broad‐spectrum resistance gene(s). DH lines were produced from F1 plants, and resistant plants were further backcrossed to produce broccoli and cauliflower‐like lines that could be useful pre‐breeding material. A combination of resistance from lines with broad and narrow specificity is recommended for controlling downy mildew in UK brassica production.

Evolutionary pathways of the pandemic influenza A (H1N1) 2009 in the UK.

The emergence of the influenza (H1N1) 2009 virus provided a unique opportunity to study the evolution of a pandemic virus following its introduction into the human population. Virological and clinical surveillance in the UK were comprehensive during the first and second waves of the pandemic in 2009, with extensive laboratory confirmation of infection allowing a detailed sampling of representative circulating viruses. We sequenced the complete coding region of the haemagglutinin (HA) segment of 685 H1N1 pandemic viruses selected without bias during two waves of pandemic in the UK (April-December 2009). Phylogenetic analysis showed that although temporal accumulation of amino acid changes was observed in the HA sequences, the overall diversity was less than that typically seen for seasonal influenza A H1N1 or H3N2. There was co-circulation of multiple variants as characterised by signature amino acid changes in the HA. A specific substitution (S203T) became predominant both in UK and global isolates. No antigenic drift occurred during 2009 as viruses with greater than four-fold reduction in their haemagglutination inhibition (HI) titre (“low reactors”) were detected in a low proportion (3%) and occurred sporadically. Although some limited antigenic divergence in viruses with four-fold reduction in HI titre might be related to the presence of 203T, additional studies are needed to test this hypothesis.

Array comparative hybridisation reveals a high degree of similarity between UK and European clinical isolates of hypervirulent Clostridium difficile

BackgroundClostridium difficile is a Gram-positive, anaerobic, spore-forming bacterium that is responsible for C. difficile associated disease in humans and is currently the most common cause of nosocomial diarrhoea in the western world. This current status has been linked to the emergence of a highly virulent PCR-ribotype 027 strain. The aim of this work was to identify regions of sequence divergence that may be used as genetic markers of hypervirulent PCR-ribotype 027 strains and markers of the sequenced strain, CD630 by array comparative hybridisation.ResultsIn this study, we examined 94 clinical strains of the most common PCR-ribotypes isolated in mainland Europe and the UK by array comparative genomic hybridisation. Our array was comprehensive with 40,097 oligonucleotides covering the C. difficile 630 genome and revealed a core genome for all the strains of 32%. The array also covered genes unique to two PCR-ribotype 027 strains, relative to C. difficile 630 which were represented by 681 probes. All of these genes were also found in the commonly occuring PCR-ribotypes 001 and 106, and the emerging hypervirulent PCR-ribotype 078 strains, indicating that these are markers for all highly virulent strains.ConclusionsWe have fulfilled the aims of this study by identifying markers for CD630 and markers associated with hypervirulence, albeit genes that are not just indicative of PCR-ribotype 027 strains. We have also extended this study and have defined a more stringent core gene set compared to those previously published due to the comprehensive array coverage. Further to this we have defined a list of genes absent from non-toxinogenic strains and defined the nature of the specific toxin deletion in the strain CD37.

23 In vitro comparison of adhesion of Candida albicans to buccal epithelial cells using Japanese and UK isolates

23 In vitro comparison of adhesion of Candida albicans to buccal epithelial cells using Japanese and UK isolates

Absence of DNA sequence diversity of the intergenic spacer 1 region inMalassezia nanaisolates from cats

Malassezia nana is a recently-described lipophilic yeast that has been isolated from the ear canals and skin of cats in Japan and Europe and from Brazilian cattle with or without otitis externa. Previous reports have demonstrated that significant intra-species variability exists in the DNA sequence of the intergenic spacer 1 region (IGS1), particularly amongst M. globosa, M. restricta and M. pachydermatis, and that certain IGS genotypes are associated with various epidemiological factors, including host disease status. In the present study, we demonstrated that the IGS1 sequences of 12 UK isolates of M. nana from cats and of six isolates from Spain (5 cat, 1 dog) were identical to each other and to CBS 9557, the M. nana type culture originally obtained from a Japanese cat with otitis externa. Further studies are needed to determine whether other genotypes of M. nana can be identified and associated with geographical regions and the species and disease status of mammalian hosts.

Is NDM-1 actually being imported to UK from India?

A recent article in Lancet Infectious Diseases suggested that Enterobacteriaceae containing New Delhi metallo-βlactamase (NDM-1) gene were being imported into UK from India. Since the study findings had widespread public health implications, it was necessary to scrutinise the adequacy of the evidence. The article was critically appraised on epidemiological, biological, and molecular evidence, the ethical principle of research, potential conflict of interest, and the justifiability of the recommendations. The study design was inappropriate to establish a causal chain between hospitalization in India and importation of NDM-1 in UK. Out of the total 29 NDM-1 positive UK patients, the NDM-1 gene was present in equal proportion among those who were hospitalized in India (44.3%) and those who were not (51.7%). Statistically significant strain relatedness between Indian and UK isolates could not be proved through dendrogram. There was a potential conflict of interest that was not disclosed. We found that the study findings did not support the authors' conclusion that India was a source for NDM-1 positive UK patients. Misplaced conclusions had the potential to cause unfounded scare and panic.

Frequency of factor H-binding protein modular groups and susceptibility to cross-reactive bactericidal activity in invasive meningococcal isolates

Meningococcal factor H-binding protein (fHbp) is a promising vaccine candidate that elicits serum bactericidal antibodies in humans. Based on sequence variability of the entire protein, fHbp has been divided into three variant groups or two sub-families. We recently reported that the fHbp architecture was modular, consisting of five variable segments, each encoded by genes from one of two lineages. Based on combinations of segments from different lineages, all 70 known fHbp sequence variants could be classified into one of six modular groups. In this study, we analyzed sequences of 172 new fHbp variants that were available from public databases. All but three variants could be classified into one of the six previously described modular groups. Among systematically collected invasive group B isolates from the U.S. and Europe, modular group I overall was most common (60%) but group IV (natural chimeras) accounted for 23% of UK isolates and <1% of U.S. isolates ( P < 0.0001). Mouse antisera to recombinant fHbp from each of the modular groups showed modular group-specific bactericidal activity against strains with low fHbp expression but had broader activity against strains with higher fHbp expression. Thus both modular group and relative expression of fHbp affected strain susceptibility to anti-fHbp bactericidal activity. The results confirmed the modular architecture of fHbp and underscored its importance for the design of broadly protective group B vaccines in different regions.

The rare oak polypore Piptoporus quercinus: Population structure, spore germination and growth

Piptoporus quercinus, the rare oak polypore, is one of only four non-lichenised fungi listed on Schedule 8 of the Wildlife and Countryside Act 1981, in the UK. It is thereby accorded the highest level of legal protection in the UK. It fruits exclusively on oak (Quercus spp.), primarily veteran trees in old growth wood pasture and parkland in the midlands and southern England. Beyond information on fruit body occurrence, little is known of the ecology of this species. Germination of chlamydospores and basidiospores was investigated under a range of conditions (temperature, pH, different media). The mating system was ascertained by pairing single spore derived isolates, and somatic compatibility investigated by pairing fruit body isolates. Germination of basidiospores was consistently below 1 % with most success occurring on 2 % malt extract agar adjusted to pH 4 at 20 °C. Chlamydospores had a much higher average percentage of germination (47 % ± 22), and germinated even after prolonged exposure to extreme conditions (desiccation or exposure to −20 °C or 40 °C for 48 h, 7 d or 14 d). The mating system was unifactorial (bipolar), although only four mating alleles were discovered in the six populations sampled, two alleles being unique to one fruit body. There were no signs of somatic incompatibility between the majority of isolates, exceptions being an isolate from Wentwood, South Wales, and an isolate from Germany collected in 1975, both of which displayed apparent signs of somatic incompatibility in all experimental pairings. The lack of somatic incompatibility between the majority of UK isolates, together with the low number of mating alleles, may point to inbreeding within the UK population. Apparently compatible isolates were not, however, clonal, as shown by variation in their mycelial extension rates on agar at various temperatures.

Susceptibility of selected freshwater fish species to a UK Lactococcus garvieae isolate

Gram-positive cocci recovered from diseased rainbow trout from a farm in England were characterized by different methods, including pulsed field gel electrophoresis, as virulent Lactococcus garvieae serogroup 2 (pulsotype A1). Groups of rainbow trout were kept at a range of temperatures and injected intraperitoneally (i.p.) with one of the UK isolates, L. garvieae 00021. The 18 degrees C and 16 degrees C groups showed 67% and 28% mortality, respectively, by day 27 post-injection. Fish kept at 14 degrees C or lower were less susceptible (< or =3% mortality). Raising the temperature of all groups to 18 degrees C at day 27 post-injection did not result in recurrence of the disease, even though viable bacteria were recovered from all groups 42 days later. Grayling were highly susceptible, with 65% mortalities when challenged with 200 colony forming unit fish(-1) by i.p. injection and 37% mortalities when exposed to effluent water from tanks containing affected rainbow trout. Other fish species tested, Atlantic salmon, brown trout and seven cyprinid species, were less susceptible. Viable L. garvieae was isolated from the internal organs of all species tested at the end of the trials, suggesting that they may pose a threat as possible carriers to susceptible farmed and wild fish.

Molecular Analysis of Korean Isolate of Barley mild mosaic virus (Iks Isolate)

The complete nucleotide sequences of both RNA of an isolated Barley mild mosaic virus (BaMMV) from Iksan, Korea, have been determined. RNA1 was 7273 nucleotides long and encodes for a polyprotein of 2261 amino acids, which contains the eight putative functional proteins. RNA2 was 3520 nucleotides long and encodes for a polyprotein of 894 amino acids, which contains two functional proteins. Results of multiple sequence alignment showed 92.9% similarity with Na1 isolate, followed by Sil, UK(F), Asl1, Remis M and UK(M) isolates, respectively. Comparison of the BaMMV-Iks polyproteins with the corresponding proteins of BaMMV-Na1 isolates showed 95% amino acid sequence identity. The phylogenetic analysis revealed that Iks isolate was closely related to Na1 strain and have a common origin.

The evaluation of novel chromogenic substrates for the detection of lipolytic activity in clinical isolates ofStaphylococcus aureusand MRSA from two European study groups

Eight novel chromogenic substrates were evaluated for their efficacy in detecting lipase activity in clinical isolates of Staphylococcus aureus from the United Kingdom and Malta. All isolates metabolized the chromogenic lipase substrates 5-(4-hydroxy-3,5-dimethoxyphenylmethylene)-2-thioxothia-zolidin-4-one-3-ethanoic acid (SRA)-propionate, SRA-butyrate, SRA-octanoate and 2-[2-(4-hydroxy-3,5-dimethoxyphenyl)-vinyl]-3-methy-benzothiazolium salt (SB(Z)TM)-acetate. Over 90% of the isolates metabolized the lipase substrates SRA-decanoate and SRA-laurate. However, only 0.6% of UK isolates and 2% of Maltese isolates metabolized the lipase substrate SRA-myristate; none of the isolates tested metabolized SB(Z)TM-butyrate. Traditional Tween 80 assays showed that over 73% of the UK methicillin-resistant Staphylococcus aureus (MRSA) isolates and 83% of the UK methicillin-sensitive Staphylococcus aureus (MSSA) isolates demonstrated lipolytic activity. In contrast, Maltese isolates showed lipase activity in 94% and 88% of the MRSA and MSSA strains, respectively. Lipases in MRSA and MSSA demonstrated substrate specificity whose activity appeared dependent upon hydrocarbon chain length of the chromogen. These novel chromogens can be used for lipase enzyme detection and have application for full characterization of numerous S. aureus lipases.

Comparison of community-associated meticillin-resistant Staphylococcus aureus from teaching hospitals in London and the USA, 2004–2006: where is USA300 in the UK?

Community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) is increasingly common in the USA, but rare in the UK. We compared CA-MRSA from the UK and USA to examine differences in the molecular epidemiology. We investigated patients presenting with MRSA in the first 72 h of hospital admission or in out-patient settings in a UK and a US hospital from January 2004 to March 2006. Fluoroquinolone susceptibility was used as a screening marker to select presumptive CA-MRSA. One hundred and eighteen and 49 such strains were identified, representing a prevalence of 0.1 and 0.2 isolates per 1,000 patient days in the UK and US respectively. Panton-Valentine leukocidin (PVL)-positive ST8-IVa (USA300)-type strains predominated among 43 surviving US isolates, whereas PVL-negative ST1-IVa predominated among 71 surviving UK isolates. There are striking differences between the molecular epidemiology of CA-MRSA in UK and US hospitals, which may have implications for control.

Absence of DNA sequence diversity of the intergenic spacer 1 region in Malassezia nana isolates from cats

Malassezia nana is a recently-described lipophilic yeast that has been isolated from the ear canals and skin of cats in Japan and Europe and from Brazilian cattle with or without otitis externa. Previous reports have demonstrated that significant intra-species variability exists in the DNA sequence of the intergenic spacer 1 region (IGS1), particularly amongst M. globosa, M. restricta and M. pachydermatis, and that certain IGS genotypes are associated with various epidemiological factors, including host disease status. In the present study, we demonstrated that the IGS1 sequences of 12 UK isolates of M. nana from cats and of six isolates from Spain (5 cat, 1 dog) were identical to each other and to CBS 9557, the M. nana type culture originally obtained from a Japanese cat with otitis externa. Further studies are needed to determine whether other genotypes of M. nana can be identified and associated with geographical regions and the species and disease status of mammalian hosts.

Ability of antibodies to two new caliciviral vaccine strains to neutralise feline calicivirus isolates from the uk

This study examined a panel of 110 uk field isolates of feline calicivirus (fcv) for susceptibility to cross-neutralisation by a panel of eight antisera raised in cats infected with fcv...

Arrival of Klebsiella pneumoniae producing KPC carbapenemase in the United Kingdom

KPC-type carbapenemases are increasingly prevalent in parts of the USA and Israel and are an emerging concern in South America, Europe and China. We investigated the UK's first two KPC-producing Klebsiella pneumoniae isolates. The isolates were referred to the UK's national reference laboratory for confirmation of carbapenem resistance. Susceptibilities were determined by agar dilution, and bla(KPC) and Tn4401-like elements were sought by PCR and sequencing. Isolates were compared by PFGE of XbaI- and SpeI-digested genomic DNA. The isolates were from patients in different UK hospitals, with no epidemiological connection. Both were resistant to carbapenems (MICs > 16 mg/L), with imipenem MICs unchanged by EDTA, and also to all other beta-lactams (including inhibitor combinations), tobramycin, amikacin and ciprofloxacin. They were susceptible to gentamicin (MICs </= 1 mg/L) and colistin (MICs </= 0.5 mg/L), with intermediate susceptibility to tigecycline (MICs 1-2 mg/L). The isolates belonged to the same PFGE-defined strain, highly related to a disseminated KPC-producing strain characterized previously in Tel Aviv, Israel. Like this Israeli strain, the UK isolates produced KPC-3 carbapenemase, with the bla(KPC-3) gene located within a Tn4401-like element. The first KPC-3-producing K. pneumoniae isolates detected in the UK were highly genetically related to a KPC-3-producing Israeli K. pneumoniae strain. This relatedness was consistent with the history of one UK patient, who had been hospitalized previously in Israel. However, this strain may be circulating more widely since the second UK patient had no identifiable links with Israel or other overseas countries.