Spread of carbapenemase-producing Morganella spp from 2013 to 2021: a comparative genomic study.

BACKGROUND: Morganella spp are opportunistic pathogens involved in various infections. Intrinsic resistance to multiple antibiotics (including colistin) combined with the emergence of carbapenemase producers reduces the number of active antimicrobials. The aim of this study was to characterise genetic features related to the spread of carbapenem-resistant Morganella spp. METHODS: This comparative genomic study included extensively drug-resistant Morganella spp isolates collected between Jan 1, 2013, and March 1, 2021, by the French National Reference Center (NRC; n=68) and European antimicrobial resistance reference centres in seven European countries (n=104), as well as one isolate from Canada, two reference strains from the Pasteur Institute collection (Paris, France), and two colistin-susceptible isolates from Bicêtre Hospital (Kremlin-Bicêtre, France). The isolates were characterised by whole-genome sequencing, antimicrobial susceptibility testing, and biochemical tests. Complete genomes from GenBank (n=103) were also included for genomic analysis, including phylogeny and determination of core genomes and resistomes. Genetic distance between different species or subspecies was performed using average nucleotide identity (ANI). Intrinsic resistance mechanisms to polymyxins were investigated by combining genetic analysis with mass spectrometry on lipid A. FINDINGS: Distance analysis by ANI of 275 isolates identified three groups: Morganella psychrotolerans, Morganella morganii subspecies sibonii, and M morganii subspecies morganii, and a core genome maximum likelihood phylogenetic tree showed that the M morganii isolates can be separated into four subpopulations. On the basis of these findings and of phenotypic divergences between isolates, we propose a modified taxonomy for the Morganella genus including four species, Morganella psychrotolerans, Morganella sibonii, Morganella morganii, and a new species represented by a unique environmental isolate. We propose that M morganii include two subspecies: M morganii subspecies morganii (the most prevalent) and M morganii subspecies intermedius. This modified taxonomy was supported by a difference in intrinsic resistance to tetracycline and conservation of metabolic pathways such as trehalose assimilation, both only present in M sibonii. Carbapenemase producers were mostly identified among five high-risk clones of M morganii subspecies morganii. The most prevalent carbapenemase corresponded to NDM-1, followed by KPC-2, and OXA-48. A cefepime-zidebactam combination was the most potent antimicrobial against the 172 extensively drug-resistant Morganella spp isolates in our collection from different European countries, which includes metallo-ß-lactamase producers. Lipid A analysis showed that the intrinsic resistance to colistin was associated with the presence of L-ARA4N on lipid A. INTERPRETATION: This global characterisation of, to our knowledge, the widest collection of extensively drug-resistant Morganella spp highlights the need to clarify the taxonomy and decipher intrinsic resistance mechanisms, and paves the way for further genomic comparisons. FUNDING: None.

Black blow fly (Diptera: Calliphoridae) bacterial symbionts inform oviposition site selection by stable flies (Diptera: Muscidae).

Larval habitats of blood-feeding stable flies, Stomoxys calcitrans (L.) (Diptera: Muscidae), overlap with foraging sites of black blow flies, Phormia regina (Meigen) (Diptera: Calliphoridae). We tested the hypothesis that bacteria in blow fly excreta inform oviposition decisions by female stable flies. In laboratory 2-choice bioassays, we offered gravid female stable flies fabric-covered agar plates as oviposition sites that were kept sterile or inoculated with either a blend of 7 bacterial strains isolated from blow fly excreta (7-isolate-blend) or individual bacterial isolates from that blend. The 7-isolate-blend deterred oviposition by female stable flies, as did either of 2 strains of Morganella morganii subsp. sibonii. Conversely, Exiguobacterium sp. and Serratia marcescens each prompted oviposition by flies. The flies' oviposition decisions appear to be guided by bacteria-derived semiochemicals as the bacteria could not be physically accessed. Oviposition deterrence caused by semiochemicals of the 7-isolate-blend may help stable flies avoid competition with blow flies. The semiochemicals of bioactive bacterial strains could be developed as trap lures to attract and capture flies and deter their oviposition in select larval habitats.

Pathometagenomics reveals susceptibility to intestinal infection by Morganella to be mediated by the blood group-related B4galnt2 gene in wild mice.

Infectious disease is widely considered to be a major driver of evolution. A preponderance of signatures of balancing selection at blood group-related genes is thought to be driven by inherent trade-offs in susceptibility to disease. B4galnt2 is subject to long-term balancing selection in house mice, where two divergent allele classes direct alternative tissue-specific expression of a glycosyltransferase in the intestine versus blood vessels. The blood vessel allele class leads to prolonged bleeding times similar to von Willebrand disease in humans, yet has been maintained for millions of years. Based on in vivo functional studies in inbred lab strains, it is hypothesized that the cost of prolonged bleeding times may be offset by an evolutionary trade-off involving susceptibility to a yet unknown pathogen(s). To identify candidate pathogens for which resistance could be mediated by B4galnt2 genotype, we here employed a novel "pathometagenomic" approach in a wild mouse population, which combines bacterial 16S rRNA gene-based community profiling with histopathology of gut tissue. Through subsequent isolation, genome sequencing and controlled experiments in lab mice, we show that the presence of the blood vessel allele is associated with resistance to a newly identified subspecies of Morganella morganii, a clinically important opportunistic pathogen. Given the increasing importance of zoonotic events, the approach outlined here may find useful application in the detection of emerging diseases in wild animal populations.

Morganella sp. infection in scent gland of corn snake (Pantherophis guttatus) - case report / Infecção por Morganella sp. em glândula de cheiro de corn snake (Pantherophis guttatus) - relato de caso

The corn snake (Pantherophis guttatus) is a non-venomous snake from the Colubridae family. The pair of scent glands is indispensable in the physiology of reproduction and defense of the species. This structure is located caudal to the cloaca. It is responsible for releasing a thick material based on lipids containing pheromones that perform functions associated with animal behavior and survival. This work aims to report infection by Morganella sp. in a scent gland in a female corn snake, evidencing the diagnosis and treatment. An increase in volume was seen in the final third, proximal to the cloaca, firm consistency, immobile, and approximately 4 cm long and painful to touch. Samples of the material were collected for bacterial culture and antibiogram examination, being positive for bacteria of the genus Morganella. Based on the result of the antibiogram, it was possible to determine the most appropriate therapeutic protocol, with the referral to perform the surgical procedure to remove the tissue compromised by the infection, remaining stable until the removal of the stitches 40 days after the procedure, with total surgical wound healing.(AU)

A corn snake (Pantherophis guttatus) é uma serpente não peçonhenta, da família Colubridae. Na fisiologia da reprodução e defesa da espécie, o par de glândulas de cheiro é indispensável. Essa estrutura está localizada caudal à cloaca e é responsável pela liberação de um material espesso à base de lipídeos, contendo feromônios que exercem funções associadas ao comportamento animal e à sua sobrevivência. O objetivo desse trabalho é relatar uma infecção por Morganella sp. em glândula de cheiro em uma fêmea de corn snake, evidenciando o diagnóstico e tratamento. Observou-se um aumento de volume no terço final, proximal a cloaca, de consistência firme, imóvel e com aproximadamente 4 cm de comprimento e sensibilidade dolorosa ao toque. Foram coletadas amostras do material para cultura bacteriana e exame de antibiograma, sendo positivo para bactérias do gênero Morganella. Com base no resultado do antibiograma, foi possível determinar o protocolo terapêutico mais adequado, como o encaminhamento para realização do procedimento cirúrgico para remoção do tecido comprometido pela infecção, se mantendo estável até a remoção dos pontos, 40 dias após o procedimento, com total cicatrização da ferida cirúrgica.(AU)

A Genomic and Bioinformatics View of the Classification and Evolution of Morganella Species and Their Chromosomal Accessory Genetic Elements Harboring Antimicrobial Resistance Genes.

In this study, draft-genome sequencing was conducted for 60 Chinese Morganella isolates, and furthermore, 12 of them were fully sequenced. Then, a total of 166 global sequenced Morganella isolates, including the above 60, were collected to perform average nucleotide identity-based genomic classification and core single nucleotide polymorphism-based phylogenomic analysis. A genome sequence-based species classification scheme for Morganella was established, and accordingly, the two conventional Morganella species were redefined as two complexes and further divided into four and two genospecies, respectively. At least 88 acquired antimicrobial resistance genes (ARGs) were disseminated in these 166 isolates and were prevalent mostly in the isolates from hospital settings. IS26/IS15DI, IS10 and IS1R, and Tn3-, Tn21-, and Tn7-subfamily unit transposons were frequently presented in these 166 isolates. Furthermore, a detailed sequence comparison was applied to 18 Morganella chromosomal accessory genetic elements (AGEs) from the fully sequenced 12 isolates, together with 5 prototype AGEs from GenBank. These 23 AGEs were divided into eight different groups belonging to composite/unit transposons, transposable prophages, integrative and mobilizable elements, and integrative and conjugative elements, and they harbored at least 52 ARGs involved in resistance to 15 categories of antimicrobials. Eleven of these 23 AGEs acquired large accessory modules, which exhibited complex mosaic structures and contained many antimicrobial resistance loci and associated ARGs. Integration of ARG-containing AGEs into Morganella chromosomes would contribute to the accumulation and dissemination of ARGs in Morganella and enhance the adaption and survival of Morganella under complex and diverse antimicrobial selection pressures. IMPORTANCE This study presents a comprehensive genomic epidemiology analysis on global sequenced Morganella isolates. First, a genome sequence-based species classification scheme for Morganella is established with a higher resolution and accuracy than those of the conventional scheme. Second, the prevalence of accessory genetic elements (AGEs) and associated antimicrobial resistance genes (ARGs) among Morganella isolates is disclosed based on genome sequences. Finally, a detailed sequence comparison of eight groups of 23 AGEs (including 19 Morganella chromosomal AGEs) reveals that Morganella chromosomes have evolved to acquire diverse AGEs harboring different profiles of ARGs and that some of these AGEs harbor large accessory modules that exhibit complex mosaic structures and contain a large number of ARGs. Data presented here provide a deeper understanding of the classification and evolution of Morganella species and also those of ARG-containing AGEs in Morganella at the genomic scale.

Impact of antibiotic susceptibility reporting on broad spectrum antibiotic use in serratia and morganella bacteremia.

To minimize broad-spectrum antibiotic use, our microbiology laboratory changed antibiotic susceptibility reporting for AmpC-beta-lactamase producing Serratia marcescens and Morganella morganii in blood cultures to include results of narrow spectrum 3rd generation cephalosporins. We assessed the impact of this change on broad-spectrum antibiotic use and clinical outcomes. All adult patients with Serratia marcescens or Morganella morganii in blood culture 2 years pre- and post-change of susceptibility reporting were retrospectively reviewed. Exclusion: more than one pathogen isolated in their blood culture, did not receive antibiotics or died within 48 hours of positive blood culture. Outcomes: Rates of broad-spectrum antibiotic use, in-hospital mortality, clinical response and microbiologic success. There were 30 patients pre-change and 46 patients post-change of reporting. Cefepime use (broad-spectrum) decreased from 46.7% to 6.5% (p < 0.001) and 3rd generation cephalosporin (narrow-spectrum) use increased (3.3% vs 34.8%, p = 0.0013) in the post-change cohort. This demonstrates the potential role of selective susceptibility reporting in antimicrobial stewardship.

Combined effects of host genetics and diet on human gut microbiota and incident disease in a single population cohort.

Human genetic variation affects the gut microbiota through a complex combination of environmental and host factors. Here we characterize genetic variations associated with microbial abundances in a single large-scale population-based cohort of 5,959 genotyped individuals with matched gut microbial metagenomes, and dietary and health records (prevalent and follow-up). We identified 567 independent SNP-taxon associations. Variants at the LCT locus associated with Bifidobacterium and other taxa, but they differed according to dairy intake. Furthermore, levels of Faecalicatena lactaris associated with ABO, and suggested preferential utilization of secreted blood antigens as energy source in the gut. Enterococcus faecalis levels associated with variants in the MED13L locus, which has been linked to colorectal cancer. Mendelian randomization analysis indicated a potential causal effect of Morganella on major depressive disorder, consistent with observational incident disease analysis. Overall, we identify and characterize the intricate nature of host-microbiota interactions and their association with disease.

An implication of biotransformation in detoxification of mercury contamination by Morganella sp. strain IITISM23.

The contamination of soil by heavy metals such as Hg is growing immensely nowadays. The drawbacks of physicochemical methods in the decontamination of polluted soils resulted in the search for an eco-friendly and cost-effective means in this regard. In this study, a potential Hg-resistant bacterial (IITISM23) strain was investigated for their removal potential of Hg, isolated from Hg-contaminated soil. IITISM23 strain was identified as Morganella sp. (MT062474.1) as it showed 99% similarity to genus Morganella of Gammaproteobacteria based on 16S rRNA gene sequencing. The toxicity experiment confirmed that the strain showed high resistance toward Hg. In low nutrient medium, EC50 (effective concentration) values were 6.8 ppm and minimum effective concentration (MIC) was 7.3 ppm, and in a nutrient-rich medium, EC50 value was 32.29 ppm and MIC value was 34.92 ppm, respectively. In in vitro conditions, IITISM23 showed the removal efficiency (81%) of Hg (II) by the volatilization method in Luria-Bertani (LB) broth. The changes in surface morphology of bacteria upon the supplementation of Hg (II) in broth media were determined by SEM-EDX studies, while the changes in functional groups were studied by FT-IR spectroscopy. The mercury reductase activity was determined by a crude extract of the bacterial strain. The optimal pH and temperature for maximum enzyme activity were 8 and 30oC, with Km of 3.5 µmol/l and Vmax of 0.88 µmol/min, respectively. Also, strain IITISM23 showed resistance toward various antibiotics and other heavy metals like cadmium, lead, arsenic, and zinc. Hence, the application of microbes can be an effective measure in the decontamination of Hg from polluted soils.

Importance of Reviewing Antibiotic Courses by 48 Hours: Risk Factors for Third-Generation Cephalosporin Resistance Among AmpC Harboring Organisms in Urine and Respiratory Cultures.

BACKGROUND: Citrobacter, Enterobacter, Morganella, and Serratia (AmpC organisms) species can exhibit third-generation cephalosporin (TGC) resistance after TGC exposure. We aimed to assess if institutional TGC utilization correlated with institutional AmpC organism susceptibility and if prior TGC exposure ≤48 hours were associated with TGC resistance in the first culture of a future infection episode caused by an AmpC organism. METHODS: A 5-year retrospective cohort study was performed, including AmpC organisms isolated from pediatric urinary and respiratory tract cultures at an institution with TGC courses reviewed by the antimicrobial stewardship program at 48 hours. Correlations were assessed by Pearson's correlation. Multivariable logistic regression identified factors independently associated with TGC resistance in a subcohort of infection episodes. RESULTS: Among 654 cultures, AmpC organism TGC susceptibility increased from 74% in 2013 to 89.3% in 2017, and this correlated with a 26.1% decrease in TGC utilization (R = -0.906; P = 0.034). Among 275 AmpC organism infections, 21.1% were resistant. Resistance occurred in 13.6%, 17.4%, and 56.5% of infections with no exposure, ≤48 hours, and >48 hours of TGC exposure in the past 30 days, respectively. TGC exposure ≤48 hours was not associated with resistance (odds ratio [OR], 1.26; 95% confidence interval [CI], 0.32-4.94; P = 0.74), whereas, TGC exposure >48 hours was (OR, 8.7; 95% CI, 3.67-20.6; P < 0.001). Infections in 2017 were less likely to be resistant (OR, 0.25; 95% CI, 0.08-0.8; P = 0.019). CONCLUSIONS: Decreased TGC utilization, likely related to antimicrobial stewardship, correlated with increased AmpC organism susceptibility. Limiting TGC exposure to ≤48 hours when possible may reduce AmpC organism resistance in future infections.

Classificação dos graus de lesões de aerossaculite em perus associadas com enterobactérias / [Classification of the levels of the airsacculitis injuries in turkeys associated with Enterobacteriaceae]

Foram padronizados os graus de lesões dos sacos aéreos em perus com aerossaculite, associadas com a presença de isolados de enterobactérias nesses órgãos. Um total de 110 amostras de sacos aéreos de perus machos com aerossaculite foi coletado para o estudo. Durante o processo de abate, as amostras foram coletadas por meio de swabs e submetidas a três métodos de armazenamento (imediato, congelado ou pré-incubado após congelamento) para posterior comparação das suas eficiências de isolamento. Os gêneros da família Enterobacteriaceae foram identificados pelas séries bioquímicas EPM, MILi e citrato de Simmons. O crescimento bacteriano ocorreu em 43,64% das amostras. Neste estudo, quatro padrões de lesões de aerossaculite foram identificados de acordo com as características patológicas dos sacos aéreos. Os principais gêneros de enterobactérias identificadas foram: Escherichia coli, Citrobacter, Proteus, Edwardsiella, Morganella, Kluyvera, Salmonella e Klebsiella. Foi observado que os graus padronizados como 3 e 4 apresentaram maior variedade de gêneros bacterianos. O armazenamento imediato apresentou maior porcentagem de positividade, 41,82%, no entanto o pré-incubado após congelamento se apresentou mais eficaz em relação à quantidade de colônias.(AU)

The degrees of air sac lesions in turkeys with airsacculitis were standardized, associated with the presence of Enterobacteriaceae isolated from these organs. A total of 110 samples of air sacs from male turkeys with airsacculitis were collected and analyzed. During the slaughtering process, the sample collection was done using swabs and submitted to three storage methods (immediate, frozen, or pre incubated after freezing) for further comparison of their isolated efficiency. The bacterial genera of the family Enterobacteriaceae were identified biochemical series EPM, MILi and Simmons citrate. Bacterial growth occurred in 43.64% of samples. In this study, four patterns of aerossaculitis lesions were identified according to the pathological characteristics of air sacs. The frequencies of the Enterobacteriaceae isolated identified in the samples were: Escherichia coli, Citrobacter, Proteus, Edwardsiella, Morganell, Kluyvera, Salmonella and Klebsiella. Otherwise, it was observed that the levels already standardized as level three and four showed higher variety of genus. The immediate storage showed higher percentage of positivity at 41.82%, however, the pre incubated after freezing showed more efficiency in relation to the quantity of colonies.(AU)

Antimicrobial susceptibility and distribution of extended-spectrum ß-lactamases, AmpC ß-lactamases and carbapenemases among Proteus, Providencia and Morganella isolated from global hospitalised patients with intra-abdominal and urinary tract infections: Results of the Study for Monitoring Antimicrobial Resistance Trends (SMART), 2008-2011.

OBJECTIVES: The increasing trend of ß-lactam resistance among Enterobacteriaceae is a worldwide problem. This study investigated isolates of the tribe Proteeae (Proteus, Providencia and Morganella) causing intra-abdominal and urinary tract infections from the worldwide Study for Monitoring Antimicrobial Resistance Trends (SMART) collected from 2008-2011. METHODS: Antimicrobial susceptibility testing was performed on isolates with an ertapenem minimum inhibitory concentration >0.5mg/L or those phenotypically producing extended-spectrum ß-lactamases (ESBLs). ESBLs, AmpC ß-lactamases and carbapenemases were detected by multiplex PCR. RESULTS: A total of 142 isolates, including Proteus mirabilis (n=121), Proteus vulgaris (n=3), Providencia stuartii (n=5), Providencia rettgeri (n=6) and Morganella morganii (n=7), were analysed. Proteus mirabilis was generally susceptible to ertapenem (∼90%) compared with imipenem (≤25%). The most common ESBLs were CTX-M types (n=64), followed by TEM (n=27) and SHV (n=7). CTX-M-1, CTX-M-2 and CTX-M-15 were the dominant CTX-M-type ESBLs in P. mirabilis isolates. CMY (n=14), which included CMY-2 (n=6), was the most common AmpC ß-lactamase, followed by DHA (n=6) and FOX (n=4). NDM (n=7), which included NDM-1 (n=4), was the most common carbapenemase, followed by KPC (n=2). Isolates from hospital-associated infections had more complicated ß-lactamase combinations than isolates from community-acquired infections. CONCLUSION: The global emergence and spread of ß-lactamase-producing Proteeae isolates are major issues in tackling antimicrobial resistance. Continuous monitoring of antimicrobial resistance trends and developing further resistance surveillance are necessary.

Histamine Production Behaviors of a Psychrotolerant Histamine-Producer, Morganella psychrotolerans, in Various Environmental Conditions.

Histamine food poisoning is a major safety concern related to seafood consumption worldwide. Morganella psychrotolerans is a novel psychrotolerant histamine-producer. In this study, the histamine production behaviors of M. psychrotolerans and two other major histamine-producers, mesophilic Morganella morganii and psychrotrophic Photobacterium phosphoreum, were compared in seafood products, and histamine accumulation by M. psychrotolerans was characterized at various pH and temperature levels in culture broth. The growth of M. psychrotolerans and P. phosphoreum increased similarly at 4 °C in canned tuna, but M. psychrotolerans produced much higher levels of histamine than P. phosphoreum. Histamine accumulation by M. psychrotolerans was induced at lower environmental pH condition at 4 and 20 °C. The optimal temperature and pH for producing histamine by crude histidine decarboxylase of M. psychrotolerans were 30 °C and pH 7, respectively. The activity of the crude HDC extracted from M. psychrotolerans cells at 10 °C retained 45% of the activity at 30 °C. Histidine decarboxylase gene expression of M. psychrotolerans was induced by low pH conditions. These results suggest that M. psychrotolerans are also a very important histamine-producer leading to histamine poisoning associated with seafood below the refrigeration temperature.

Understanding the role of SilE in the production of metal nanoparticles by Morganella psychrotolerans using MicroScale Thermophoresis.

Metal nanoparticle synthesis has been observed in several species of bacteria but the underlying mechanisms of synthesis are not well understood. Morganella psychrotolerans is a Gram-negative psychrophilic bacterium that is able to tolerate relatively high concentrations of Cu and Ag ions, and it is through the associated resistance pathways that this species is able to convert metal ions to nanoparticles. The purpose of this study was to investigate the mechanism of nanoparticle synthesis, looking at the interaction of the metal binding protein SilE with metal ions using MicroScale Thermophoresis (MST). MST assays give a rapid and accurate determination of binding affinities, allowing for the testing of SilE with a range of environmentally significant metal ions. The binding affinities (Kd) of Ag+ and Cu2+ were measured as 0.17 mM and 0.13 mM respectively, consistent with the observations of strong binding reported in the literature, whereas the binding to Al3+ and Co2+ was measured as Kd values of 4.19 mM and 1.35 mM respectively.

High prevalence of multiple drug resistant enteric bacteria: Evidence from a teaching hospital in Southwest Nigeria.

The development and evolution of antimicrobial resistance (AMR) in pathogens has been reported to be one of the major issues confronting the global health community. The aim of this study was to examine the period prevalence of antibiotic resistance, as well as the trends and patterns in sensitivity profile of enteric bacteria isolated from urine samples of patients with UTIs in a teaching Hospital in south west Nigeria. Urine samples were collected from 77 patients with UTIs from February 2017 to October 2018. Standard laboratory methods were used for urine sample culture and bacterial identification. The Kirby-Bauer disk diffusion method was used to evaluate antimicrobial sensitivity. Predominant enteric bacteria isolates were Escherichia coli (24, 39.3%), Salmonella species (12, 19.7%), Klebsiella species (4, 6.6%), Providencia species (6, 9.8%), Proteus species (8, 13.1%), Serratia species (2, 3.3%), Yersinia species (1, 1.6%) and Morganella species (4, 6.6%). A large proportion (90.2%) of isolates obtained were multi-drug resistant. High resistance in amoxycillin-clavulanate (98%), cefuroxime (92%), erythromycin (90%) and ceftazidime (84%) were recorded. These results emphasize the importance of continuous screening and surveillance programmes for detection of AMR in enteric bacteria of public health importance.

Proteae: a reservoir of class 2 integrons?

OBJECTIVES: Our aim was to confirm with a large panel of clinical isolates that class 2 integrons are highly prevalent in Proteae and to analyse their genetic characteristics. METHODS: Proteae (Proteus spp., Morganella spp. and Providencia spp.) isolates were collected from clinical samples during 2013 at Limoges University Hospital, France. The presence of class 1, 2 and 3 integrons was investigated by quantitative PCR. The presence of a stop codon in the intI2 gene was determined by Sanger sequencing. The gene cassette arrays of class 2 integrons were determined by PCR-RFLP and Sanger sequencing or next-generation sequencing when needed. RESULTS: Of the 327 Proteae collected, 103 (31.5%) harboured a class 2 integron and 45 (13.8%) a class 1 integron. No class 3 integrons were detected. One functional IntI2 integrase was detected in a Morganella morganii isolate. Six different gene cassette arrays were detected. Four had already been described in the literature: dfrA1-sat2-aadA1 (72 isolates), dfrA1-catB2-sat2-aadA1 (17), sat2-aadA1 (6) and lnu(F), dfrA1, aadA1 (1). We identified two new gene cassette arrays: (i) a new variant of the dfrA1 gene cassette (one isolate; the one with the functional IntI2); and (ii) the array dfrA1-gcu115-sat2 harbouring the new gcu115 gene cassette with two ORFs encoding proteins of unknown functions (five isolates). CONCLUSIONS: We showed a high frequency of class 2 integrons, as well as a diversity of gene cassette arrays, among Proteae. This work highlights that the Proteae tribe plays an important role as a reservoir of class 2 integrons.

Morganella as a cause of ophthalmia neonatorum.

A 2 day-old-girl with a complicated birth history, including prolonged rupture of membranes and Caesarian section delivery, presented with a purulent bacterial conjunctivitis of the left eye despite standard prophylaxis. Bacterial cultures yielded 1+ Morganella morganii, a facultative anaerobe and a rare cause of ocular pathology. The patient was treated with topical tobramycin four times daily for 7 days, resulting in resolution of her conjunctivitis.

Room temperature bioproduction, isolation and anti-microbial properties of stable elemental copper nanoparticles.

In nanoparticle production there are a number of important considerations that must be made. Producing nanoparticles of uniform size and shape is vital, but no less important is ensuring the production process is as efficient as possible in time, cost and energy. Traditional chemical and physical methods of nanoparticle production often involve high temperatures and pressures, as well as the use of toxic substrates; in contrast the bioproduction of nanoparticles is greener and requires a smaller input of energy resources. Here we outline a method for the straightforward bioproduction of stable, uniform elemental (zero-valent) copper nanoparticles at room temperature, and demonstrate how their size and shape can be modified by subsequent pH manipulation. We also highlight a potential application for these biogenic copper nanoparticles by demonstrating their potential to inhibit bacterial growth.

Characterization and genomic analyses of two newly isolated Morganella phages define distant members among Tevenvirinae and Autographivirinae subfamilies.

Morganella morganii is a common but frequent neglected environmental opportunistic pathogen which can cause deadly nosocomial infections. The increased number of multidrug-resistant M. morganii isolates motivates the search for alternative and effective antibacterials. We have isolated two novel obligatorily lytic M. morganii bacteriophages (vB_MmoM_MP1, vB_MmoP_MP2) and characterized them with respect to specificity, morphology, genome organization and phylogenetic relationships. MP1's dsDNA genome consists of 163,095 bp and encodes 271 proteins, exhibiting low DNA (<40%) and protein (<70%) homology to other members of the Tevenvirinae. Its unique property is a >10 kb chromosomal inversion that encompass the baseplate assembly and head outer capsid synthesis genes when compared to other T-even bacteriophages. MP2 has a dsDNA molecule with 39,394 bp and encodes 55 proteins, presenting significant genomic (70%) and proteomic identity (86%) but only to Morganella bacteriophage MmP1. MP1 and MP2 are then novel members of Tevenvirinae and Autographivirinae, respectively, but differ significantly from other tailed bacteriophages of these subfamilies to warrant proposing new genera. Both bacteriophages together could propagate in 23 of 27 M. morganii clinical isolates of different origin and antibiotic resistance profiles, making them suitable for further studies on a development of bacteriophage cocktail for potential therapeutic applications.

Development of a real-time PCR method coupled with a selective pre-enrichment step for quantification of Morganella morganii and Morganella psychrotolerans in fish products.

Histamine fish poisoning is common and due to toxic concentrations of histamine often produced by Gram-negative bacteria in fin-fish products with a high content of the free amino acid histidine. The genus Morganella includes two species previously reported to cause incidents of histamine fish poisoning. Morganella morganii and Morganella psychrotolerans are both strong producer of histamine. However, little is known about the occurrence and critical stages for fish contamination with these bacteria. To elucidate contamination routes of Morganella, specific real-time quantitative PCR (RTi qPCR) methods for quantification of M. morganii and M. psychrotolerans have been developed. Selective primers amplified a 110 bp region of the vasD gene for M. psychrotolerans and a 171 bp region of the galactokinase gene for M. morganii. These primer-sets showed high specificity as demonstrated by using purified DNA from 23 other histamine producing bacteria and 26 isolates with no or limited histamine production. The efficiency of the qPCR reactions on artificially contaminated fish samples were 100.8% and 96.3% respectively. The limit of quantification (LOQ) without enrichment was 4 log CFU/g. A quantitative enrichment step with a selective medium was included and improved the sensitivity of the methods to a LOQ of below 50 CFU/g in seafood. RTi qPCR methods with or without enrichment were evaluated for enumeration of Morganella species in naturally contaminated fresh fish and lightly preserved seafood from Denmark. These new methods will contribute to a better understanding of the occurrence and histamine production by Morganella species in fish products, information that is essential to reduce the unacceptably high frequency of histamine fish poisoning.