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The need for alternatives to antibiotic therapy due to the emergence of multidrug resistant bacteria (MDR), such as the nosocomial pathogen Acinetobacter baumannii, has led to the recovery of phage therapy. In addition, phages can be combined in cocktails to increase the host range. In this study, the evolutionary mechanism of adaptation was utilized in order to develop a phage adapted to A. baumannii, named phage Ab105-2phiCI404ad, from a mutant lytic phage, Ab105-2phiCI, previously developed by our group. The whole genome sequence of phage Ab105-2phiCI404ad was determined, showing that four genomic rearrangements events occurred in the tail morphogenesis module affecting the ORFs encoding the host receptor binding sites. As a consequence of the genomic rearrangements, 10 ORFs were lost and four new ORFs were obtained, all encoding tail proteins; two inverted regions were also derived from these events. The adaptation process increased the host range of the adapted phage by almost three folds. In addition, a depolymerase-expressing phenotype, indicated by formation of a halo, which was not observed in the ancestral phage, was obtained in 81\% of the infected strains. A phage cocktail was formed by combining this phage with the A. baumannii phage vB\_AbaP\_B3, known to express a depolymerase. Both the individual phages and the phage cocktail showed strong antimicrobial activity against 5 clinical strains and 1 reference strain of A. baumannii tested. However, in all cases resistance to the bacterial strains was also observed. The antibiofilm activity of the individual phages and the cocktail was assayed. The phage cocktail displayed strong antibiofilm activity., This study was funded by grants PI16/01163 and PI19/00878 awarded to M. Tomás within the State Plan for R+D+I 2013-2016 (National Plan for Scientific Research, Technological Development and Innovation 2008-2011) and co-financed by the ISCIII 471 Deputy General Directorate for Evaluation and Promotion of Research - European Regional Development Fund "A way of Making Europe" and Instituto de Salud Carlos III FEDER, Spanish Network for the Research in Infectious Diseases (REIPI, RD16/0016/0001, RD16/0016/0006 and RD16/CIII/0004/0002) and by the Study Group on Mechanisms of Action and Resistance to Antimicrobials, GEMARA (SEIMC, http://www.seimc.org/). M. Tomás was financially supported by the Miguel Servet Research Programme (SERGAS and ISCIII). I. Bleriot was financially supported by pFIS program (ISCIII, FI20/00302). O. Pacios and M. López were financially supported by 4grants IN606A-2020/035 and IN606B-2018/008, respectively (GAIN, Xunta de Galicia)., info:eu-repo/semantics/publishedVersion

Ventilator-associated pneumonia (VAP) due to Acinetobacter spp. is one of the most common infections in the intensive care unit. Hence, we performed this prospective-observational multicenter study, and described the course and outcome of the disease. This study was performed in 24 centers between January 06, 2014, and December 02, 2016. The patients were evaluated at time of pneumonia diagnosis, when culture results were available, and at 72 h, at the 7th day, and finally at the 28th day of follow-up. Patients with coexistent infections were excluded and only those with a first VAP episode were enrolled. Logistic regression analysis was performed. A total of 177 patients were included; empiric antimicrobial therapy was appropriate (when the patient received at least one antibiotic that the infecting strain was ultimately shown to be susceptible) in only 69 (39%) patients. During the 28-day period, antibiotics were modified for side effects in 27 (15.2%) patients and renal dose adjustment was made in 38 (21.5%). Ultimately, 89 (50.3%) patients died. Predictors of mortality were creatinine level (OR, 1.84 (95% CI 1.279–2.657); p = 0.001), fever (OR, 0.663 (95% CI 0.454–0.967); p = 0.033), malignancy (OR, 7.095 (95% CI 2.142–23.500); p = 0.001), congestive heart failure (OR, 2.341 (95% CI 1.046–5.239); p = 0.038), appropriate empiric antimicrobial treatment (OR, 0.445 (95% CI 0.216–0.914); p = 0.027), and surgery in the last month (OR, 0.137 (95% CI 0.037–0.499); p = 0.003). Appropriate empiric antimicrobial treatment in VAP due to Acinetobacter spp. was associated with survival while renal injury and comorbid conditions increased mortality. Hence, early diagnosis and appropriate antibiotic therapy remain crucial to improve outcomes. © 2019, Springer-Verlag GmbH Germany, part of Springer Nature.

BackgroundThe enzymatic degradation of quorums sensing (QS) molecules (called quorum quenching, QQ) has been considered as a promising anti-virulence therapy to treat biofilm-related infections and combat antibiotic resistance. The recently-discovered QQ enzyme MomL has been reported to efficiently degrade differentN-acyl homoserine lactones (AHLs) of various Gram-negative pathogens. Here we investigated the effect of MomL on biofilms formed by two important nosocomial pathogens,Pseudomonas aeruginosaandAcinetobacter baumannii.MethodsMomL was expressed inE.coliBL21 and purified. The activity of MomL on AHLs with hydroxyl substituent was tested. Biofilms ofP. aeruginosaPAO1 andAcinetobacterstrains were formed in 96-well microtiter plates. Biofilm formation was evaluated by crystal violet staining, plating and fluorescence microscopy. The effect of MomL on biofilm susceptibility to antibiotics was also tested. We further evaluated MomL in dual-species biofilms formed byP. aeruginosaandA. baumannii, and in biofilms formed in a wound model. The effect of MomL on virulence ofA. baumanniiwas also tested in theCaenorhabditis elegansmodel.ResultsMomL reduced biofilm formation and increased biofilm susceptibility to different antibiotics in biofilms ofP. aeruginosaPAO1 andA. baumanniiLMG 10531 formed in microtiter platesin vitro. However, no significant differences were detected in the dual-species biofilm and in wound model biofilms. In addition, MomL did not affect virulence ofA. baumanniiin theC. elegansmodel. Finally, the effect of MomL on biofilm ofAcinetobacterstrains seems to be strain-dependent.DiscussionOur results indicate that although MomL showed a promising anti-biofilm effect againstP. aeruginosaandA. baumaniibiofilms formed in microtiter plates, the effect on biofilm formation under conditions more likely to mimic the real-life situation was much less pronounced or even absent. Our data indicate that in order to obtain a better picture of potential applicability of QQ enzymes for the treatment of biofilm-related infections, more elaborate model systems need to be used.

The structure of the surface polysaccharide from a hypervirulent for mice Acinetobacter baumannii strain LAC-4 was studied. The polysaccharide was built of trisaccharide repeating units containing α-l-fucosamine, α-d-glucosamine, and α-8-epi-legionaminic acid. The structure interpretation was based mostly on NMR data. Polysaccharide was obtained using a procedure of LPS O-chain preparation, although whether it is an LPS O-chain or capsular polysaccharide remained unclear. © 2014 Published by Elsevier Ltd. All rights reserved.

Acinetobacter spp. are found in 53% of air colonization samples from the hospital environment. In this work, we sequenced all the genome of airborne Acinetobacter sp. strain 5-2Ac02. We found important features at the genomic level in regards to the rhizome. By phylogenetic analysis, A. towneri was the species most closely related to Acinetobacter sp. 5-2Ac02.

Acinetobacter ursingii is an aerobic, gram-negative, opportunistic microorganism which is rarely isolated among Acinetobacter species. We present two immunocompetent infants who developed bacteremia due to A.ursingii. The first patient is a two -month- old boy who had been hospitalized in pediatric surgery unit for suspected tracheo-esophageal fistula because of recurrent aspiration pneumonia unresponsive to antibiotic therapy. The second patient is a fourteen -month- old boy with prolonged vomiting and diarrhea. A. ursingii was isolated from their blood cultures. They were successfully treated with ampicillin-sulbactam. Although A.ursingii has recently been isolated from a clinical specimen; reports of infection with A.ursingii in children are rare. A.ursingii should be kept in mind as an opportunistic microorganism in children.Pan African Medical Journal 2016; 23

Made available in DSpace on 2019-09-12T16:53:35Z (GMT). No. of bitstreams: 0 Previous issue date: 2015 Merieux Research Grants - Institut Merieux We evaluated the epidemiology of Acinetobacter spp. recovered from patients diagnosed with bloodstream infections in 9 tertiary hospitals located in all Brazilian geographic regions between April and August 2014. Although OXA-23-producing Acinetobacter baumannii clones were disseminated in most hospitals, it was observed for the first time the spread of OXA-72 among clonally related A. baumannii isolated from distinct hospitals. Interestingly, Acinetobacter pittii was the most frequent species found in a Northern region hospital. Contrasting with the multisusceptible profile displayed by A. pittii isolates, the tetracyclines and polymyxins were the only antimicrobials active against all A. baumannii isolates. (C) 2015 Elsevier Inc. All rights reserved. [Vasconcelos, Ana Tereza R.] Lab Nacl Comp Cient LNCC MCTI, Petropolis, RJ, Brazil [Barth, Afonso L.; Zavascki, Alexandre P.] Univ Fed Rio Grande do Sul, Hosp Clin Porto Alegre, Lab Pesquisa Resistencia Bacteriana LABRESIS, Porto Alegre, RS, Brazil [Gales, Ana C.; Furtado, Guilherme H. C.; da Silva, Juliana O.; Correa, Luci; Cayo, Rodrigo; Martins, Willames M. B. S.] Univ Fed Sao Paulo UNIFESP, Disciplina Infectol, Dept Med, Sao Paulo, SP, Brazil [Levin, Anna S.; Rossi, Flavia; Silva, Mariama T.; Oliveira, Maura S.; Dalben, Mirian F.; Santos, Sania A.] Univ Sao Paulo, Inst Med Trop USP LIM 54, Dept Patol LIM 03, Hosp Clin,Fac Med, Sao Paulo, SP, Brazil [Lucarevschi, Bianca R.; Moreira, Marina] Universidade de Taubaté (Unitau) , Dept Med [Cabral, Blenda G.; Brasiliense, Danielle M.; Carneiro, Irna Carla R. S.; Lima, Karla V. B.; da Conceicao, Marilia L.] Fundacao Santa Casa Misericordia Para UFPA, Belem, Para, Brazil [Cabral, Blenda G.; Brasiliense, Danielle M.; Carneiro, Irna Carla R. S.; Lima, Karla V. B.; da Conceicao, Marilia L.] Inst Evandro Chagas SVS MS, Belem, Para, Brazil [Ribeiro, Julival; Guzman, Ricardo D.] Hosp Base, Brasilia, DF, Brazil [Correa, Luci; Martino, Marines D. V.] Hosp Israelita Albert Einstein HIAE, Sao Paulo, SP, Brazil [Correa, Luci; Martino, Marines D. V.] Fac Ciencias Med Santa Casa Sao Paulo, Sao Paulo, SP, Brazil [Britto, Maria H.; de Freitas, Manse R.; Morais, Rosangela] Univ Fed Rio Grande Norte UFRN, Ctr Patol Clin, Natal, RN, Brazil

We previously showed that the opportunistic nosocomial pathogen Acinetobacter baumannii is able to sense and respond to light via BlsA, a BLUF (Blue-Light-sensing Using FAD)-domain photoreceptor protein. Here, we extend our previous studies showing that light regulation is not restricted to A. baumannii, but rather widespread within the genus Acinetobacter. First, we found that blue light modulates motility and biofilm formation in many species of the genus, including members of the Acinetobacter calcoaceticus-A. baumannii complex. In many of these species blue light acts as a key factor guiding the decision between motility or sessility at 24uC, whereas in A. baumannii, light inhibits both motility and biofilm formation. We also show that light regulation of motility occurred not only at 24uC but also at 37uC in non-A. baumannii species, contrasting the situation of A. baumannii which only shows photoregulation at 24uC. Second, we show that Acinetobacter baylyi (strain ADP1) BLUF-photoreceptors can functionally replace in vivo the A. baumannii 17978 BlsA protein and that the pathways leading to biofilm formation are inversely regulated at 24uC between these two microorganisms. Finally, we found the presence of predicted genes coding BLUF-containing proteins in all Acinetobacter sequenced genomes, even though the copy number is variable among them. Phylogenetic analysis suggests a common origin for all BLUF domains present in members of this genus, and could distinguish well-differentiated clusters that group together BLUF homologs from different species, a situation particularly clear for members of the ACB complex. Despite a role played by these BLUF domain-containing proteins in the photoregulation observed in the members of the genus Acinetobacter is a likely scenario given our findings in A. baumannii and A. baylyi, further research will contribute to confirm this possibility. Fil: Golic, Adrián Ezequiel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Instituto de Biología Molecular y Celular de Rosario; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Centro de Estudios Fotosintéticos y Bioquímicos (i); Argentina Fil: Vaneechoutte, Mario. Ghent University. Ghent University Hospital. Laboratory for Bacteriology Research; Bélgica Fil: Nemec, Alexandr. National Institute of Public Health. Laboratory of Bacterial Genetics; República Checa Fil: Viale, Alejandro Miguel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Instituto de Biología Molecular y Celular de Rosario; Argentina Fil: Actis, Luis A.. Miami University. Department of Microbiology; Estados Unidos Fil: Mussi, María Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Centro de Estudios Fotosintéticos y Bioquímicos (i); Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Instituto de Biología Molecular y Celular de Rosario; Argentina

Background Prokaryotic plasmids have a dual importance in the microbial world: first they have a great impact on the metabolic functions of the host cell, providing additional traits that can be accumulated in the cell without altering the gene content of the bacterial chromosome. Additionally and/or alternatively, from a genome perspective, plasmids can provide a basis for genomic rearrangements via homologous recombination and so they can facilitate the loss or acquisition of genes during these events, which eventually may lead to horizontal gene transfer (HGT). Given their importance for conferring adaptive traits to the host organisms, the interest in plasmid sequencing is growing and now many complete plasmid sequences are available online. Results By using the newly developed Blast2Network bioinformatic tool, a comparative analysis was performed on the plasmid and chromosome sequence data available for bacteria belonging to the genus Acinetobacter, an ubiquitous and clinically important group of γ-proteobacteria. Data obtained showed that, although most of the plasmids lack mobilization and transfer functions, they have probably a long history of rearrangements with other plasmids and with chromosomes. Indeed, traces of transfers between different species can be disclosed. Conclusions We show that, by combining plasmid and chromosome similarity, identity based, network analysis, an evolutionary scenario can be described even for highly mobile genetic elements that lack extensively shared genes. In particular we found that transposases and selective pressure for mercury resistance seem to have played a pivotal role in plasmid evolution in Acinetobacter genomes sequenced so far.

The taxonomic status of 24 haemolytic, non-glucose acidifying Acinetobacter strains that did not belong to any previously described species was investigated by means of a polyphasic approach. Using AFLP fingerprinting, amplified rDNA restriction analysis and phenotypic characterization, the strains were classified into two phenetically coherent groups (comprising 15 and 9 strains) that were distinct from each other and from all known Acinetobacter species. Confirmation that these groups formed two separate lineages within the genus Acinetobacter was obtained from comparative analysis of partial sequences of the gene encoding the beta-subunit of RNA polymerase in all strains and also from 16S rRNA gene sequence analysis of representative strains. Previously published DNA-DNA reassociation data for some of the strains used also supported the species rank for both groups, for which the names Acinetobacter beijerinckii sp. nov. and Acinetobacter gyllenbergii sp. nov. are proposed. The strains of A. beijerinckii sp. nov. originated from human and animal specimens and from various environmental sources, whereas those of A. gyllenbergii sp. nov. were isolated exclusively from human clinical specimens. The phenotypic characteristics most useful for the differentiation of these species from other Acinetobacter species that comprise haemolytic strains were the inability of A. beijerinckii sp. nov. to grow on L-arginine and the ability of A. gyllenbergii sp. nov. to grow on azelate. The type strain of A. beijerinckii sp. nov. is NIPH 838(T) (=LUH 4759(T)= CCUG 51249(T)= CCM 7266(T) =58a(T)) and the type strain of A. gyllenbergii sp. nov. is NIPH 2150(T) (=RUH 422(T)= CCUG 51248(T)=CCM 7267(T)=1271(T)).

The gram-negative bacterium Acinetobacter baumannii strain ATCC17961 has been used by several laboratories in mouse models of respiratory A. baumannii infection, and a study of the role of its lipopolysaccharide in the pathogenicity is of interest. The structure of the O-deacylated polysaccharide O-chain component of its LPS has been determined by 2D NMR spectroscopy and mass spectrometry methods, and by the structural identification of oligosaccharides obtained by sequential application of the Smith degradation of the O-antigen. The O-chain was determined to be a polymer of a branched pentasaccharide repeating unit composed of 2,3-diacetamido-2,3-dideoxy-D-glucuronic acid, 2-acetamido-2-deoxy-D-glucose, 2-acetamido-2-deoxy-D-galactose, D-glucose, and D-galactose, and has the following structure: [carbohydrate sequence see in text].

Recently, an extended-spectrum beta -lactamase (PER-I) was found to be disseminated among Acinetobacter spp, and Pseudomonas aeruginosa isolates in Turkey. A population-based cohort study was conducted to elucidate predictive mortality factors in patients with nosocomial infections caused by Acinetobacter spp. and P. aeruginosa, with particular reference to PER-1-type extended-spectrum beta -lactamase (ESBL) production. The study group comprised 16 and 21 non-survivors and 82 and 126 survivors in cohorts infected with Acinetobacter and E. aeruginosa, respectively. In the Acinetobacter-infected cohort, nosocomial pneumonia, hypotension and infection with a PER-positive isolate were independent predictors of mortality. In the P. aeruginosa-infected cohort, impaired consciousness, a PER-positive isolate, male sex and (with a negative relative risk) urinary tract infection were independent predictors of death. This study demonstrated the relationship of PER-1-type ESBL-producing Acinetobacter spp. and P. aeruginosa with poor clinical outcome.