Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 158
Filtrar
Mais filtros

Intervalo de ano de publicação
1.
Nat Chem Biol ; 2024 Jun 19.
Artigo em Inglês | MEDLINE | ID: mdl-38898166

RESUMO

Bacterial infection involves a complex interaction between the pathogen and host where the outcome of infection is not solely determined by pathogen eradication. To identify small molecules that promote host survival by altering the host-pathogen dynamic, we conducted an in vivo chemical screen using zebrafish embryos and found that treatment with 3-hydroxykynurenine (3-HK) protects from lethal bacterial infection. 3-HK, a metabolite produced through host tryptophan metabolism, has no direct antibacterial activity but enhances host survival by restricting bacterial expansion in macrophages through a systemic mechanism that targets kainate-sensitive glutamate receptors. These findings reveal a new pathway by which tryptophan metabolism and kainate-sensitive glutamate receptors function and interact to modulate immunity, with important implications for the coordination between the immune and nervous systems in pathological conditions.

2.
J Immunol ; 213(5): 619-627, 2024 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-39037267

RESUMO

Sepsis is a complex condition of inflammatory and immune dysregulation, triggered by severe infection. In survivors, chronic inflammation and immune dysregulation linger, facilitating the emergence of infections. CD8 dysfunction contributes to immunosuppression in sepsis survivors. We devised an animal model that enabled us to identify and analyze CD8-intrinsic defects induced by sepsis. We adoptively transferred CD45.1 CD8 OT-I T cells into CD45.2 congenic mice and subjected them to cecal ligature and puncture, to induce abdominal sepsis. One month later, we isolated the transferred CD8 cells. Surface marker expression confirmed they had not been activated through the TCR. CD8 OT-I T cells isolated from septic (or sham-operated) mice were transferred to second recipients, which were challenged with OVA-expressing Listeria monocytogenes. We compared effector capacities between OT-I cells exposed to sepsis and control cells. Naive mice that received OT-I cells exposed to sepsis had higher bacterial burden and a shorter survival when challenged with OVA-expressing L. monocytogenes. OT-I cells isolated from septic mice produced less IFN-γ but had conserved activation, expansion potential, and cytotoxic function. We observed lower transcript levels of IFN-γ and of the long noncoding RNA Ifng-as1, a local regulator of the epigenetic landscape, in cells exposed to sepsis. Accordingly, local abundance of a histone modification characteristic of active promoter regions was reduced in sepsis-exposed CD8 T cells. Our results identify a mechanism through which inflammation in the context of sepsis affects CD8 T cell function intrinsically.


Assuntos
Linfócitos T CD8-Positivos , Cromatina , Interferon gama , Listeria monocytogenes , Sepse , Animais , Sepse/imunologia , Linfócitos T CD8-Positivos/imunologia , Camundongos , Interferon gama/imunologia , Cromatina/imunologia , Cromatina/metabolismo , Listeria monocytogenes/imunologia , Camundongos Endogâmicos C57BL , Transferência Adotiva , Modelos Animais de Doenças , Listeriose/imunologia , Ativação Linfocitária/imunologia
3.
Int J Mol Sci ; 23(9)2022 Apr 25.
Artigo em Inglês | MEDLINE | ID: mdl-35563122

RESUMO

The marine bacterium Photobacterium damselae subsp. piscicida (Pdp) causes photobacteriosis in fish and important financial losses in aquaculture, but knowledge of its virulence factors is still scarce. We here demonstrate that an unstable plasmid (pPHDPT3) that encodes a type III secretion system (T3SS) is highly prevalent in Pdp strains from different geographical origins and fish host species. We found that pPHDPT3 undergoes curing upon in vitro cultivation, and this instability constitutes a generalized feature of pPHDPT3-like plasmids in Pdp strains. pPHDPT3 markers were detected in tissues of naturally-infected moribund fish and in the Pdp colonies grown directly from the fish tissues but were undetectable in a fraction of the colonies produced upon the first passage of the primeval colonies on agar plates. Notably, cured strains exhibited a marked reduction in virulence for fish, demonstrating that pPHDPT3 is a major virulence factor of Pdp. The attempts to stabilize pPHDPT3 by insertion of antibiotic resistance markers by allelic exchange caused an even greater reduction in virulence. We hypothesize that the existence of a high pressure to shed pPHDPT3 plasmid in vitro caused the selection of clones with off-target mutations and gene rearrangements during the process of genetic modification. Collectively, these results show that pPHDPT3 constitutes a novel, hitherto unreported virulence factor of Pdp that shows a high instability in vitro and warn that the picture of Pdp virulence genes has been historically underestimated, since the loss of the T3SS and other plasmid-borne genes may have occurred systematically in laboratories for decades.


Assuntos
Doenças dos Peixes , Infecções por Bactérias Gram-Negativas , Animais , Doenças dos Peixes/microbiologia , Peixes/genética , Infecções por Bactérias Gram-Negativas/microbiologia , Photobacterium/genética , Plasmídeos/genética , Sistemas de Secreção Tipo III/genética , Virulência/genética , Fatores de Virulência/genética
4.
Rev Med Chil ; 150(1): 100-106, 2022 Jan.
Artigo em Espanhol | MEDLINE | ID: mdl-35856971

RESUMO

Dr. Vicente Izquierdo San Fuentes was the first professor of Histology at the Faculty of Medicine of the University of Chile. In that Chair, cell theory strongly radiated to new generations of health students. However, the conditions for the creation of the discipline of General or Cell Biology were not yet ripe. Almost three decades later, Dr. Juan Noé Crevani was hired in Italy to lead Medical Zoology in 1912. From the heterogeneous discipline of Medical Zoology, Dr. Noé managed to create in 1926 the new chairs of General Biology, Embryology-Comparative Anatomy and Parasitology. His vision of biology as an essentially dynamic and experimental science, contributed to modernize and encourage the development of different areas of biology in Chile. Retaining their full independence, these chairs met in 1931, in a new organization called the Juan Noé Institute of Biology, which lasted until the university reform of 1968. Afterwards, the departments of Biology and Genetics, Parasitology, Human Anatomy and Histology were created. In 1998, a new reorganization of the Faculty of Medicine of the University of Chile began, creating the so-called Institute of Biomedical Sciences (ICBM) that houses several disciplinary programs that replaced the old departments.


Assuntos
Docentes , Medicina , Academias e Institutos , Chile , História do Século XX , Humanos , Universidades/história
5.
Environ Microbiol ; 23(9): 4859-4880, 2021 09.
Artigo em Inglês | MEDLINE | ID: mdl-34423883

RESUMO

The marine bacterium Photobacterium damselae subsp. damselae (Pdd) causes disease in marine animals and humans. Previous studies demonstrated that mutation of the two-component system RstAB strongly impacts virulence of this pathogen, but the RstAB regulon has not been thoroughly elucidated. We here compared the transcriptomes of Pdd RM-71 and ΔrstA and ΔrstB derivatives using RNA-seq. In accordance with previous studies, RstAB positively regulated cytotoxins Dly, PhlyP and PhlyC. This analysis also demonstrated a positive regulation of outer membrane proteins, resistance against antimicrobials and potential virulence factors by this system. Remarkably, RstAB positively regulated two hitherto uncharacterised gene clusters involved in the synthesis of a polysaccharide capsule. Presence of a capsular layer in wild-type cells was confirmed by transmission electron microscopy, whereas rstA and rstB mutants were non-capsulated. Mutants for capsule synthesis genes, wza and wzc exhibited acapsular phenotypes, were impaired in resistance against the bactericidal action of fish serum and mucus, and were strongly impaired in virulence for fish, indicating a major role of capsule in virulence. Collectively, this study demonstrates that RstAB is a major positive regulator of key virulence factors including a polysaccharide capsule essential for full virulence in a pathogenic Photobacterium.


Assuntos
Doenças dos Peixes , Photobacterium , Animais , Humanos , Photobacterium/genética , Polissacarídeos , Virulência/genética
6.
Microb Ecol ; 80(3): 507-518, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32385615

RESUMO

The marine bacterium Photobacterium damselae subsp. damselae is a pathogen that causes disease in diverse marine animals, and is also a serious opportunistic human pathogen that can cause fatal infections. Strains of this pathogen isolated from diseased European sea bass in aquaculture facilities in the Turkish coast of the Black Sea were found to exhibit reduced sensitivity to multiple antimicrobials. Selected representative strains were subjected to complete genome sequencing and plasmid characterization. It was found that multidrug resistant (MDR) isolates harboured large conjugative plasmids sharing part of their sequence backbone with pAQU-group plasmids, hitherto reported exclusively in China and Japan. Four new pAQU-group versions of plasmids were identified in the present study, containing distinct combinations of the resistance determinants tetB, floR, sul2, qnrVC, dfrA and strAB. Conjugative transfer of pPHDD2-OG2, a representative plasmid of 170,998 bp, occurred at high frequencies (2.2 × 10-2 transconjugants per donor cell), to E. coli and to pathogenic P. damselae subsp. damselae and subsp. piscicida strains. Upon transfer, pPHDD2-OG2 conferred reduced susceptibility to a number of antimicrobials to the recipient strains. Comparative genomics analysis of host strains suggested that these MDR plasmids of the pAQU-group were acquired by different genetic lineages of Pdd. This study provides evidence that P. damselae subsp. damselae isolated from diseased fish constitute a reservoir for conjugative MDR pAQU-group plasmids in the Mediterranean basin, and have the potential to spread to diverse bacterial species.


Assuntos
Farmacorresistência Bacteriana Múltipla/genética , Photobacterium/genética , Plasmídeos/genética , Aquicultura , Mar Negro
7.
J Bacteriol ; 200(15): e00002-18, 2018 02 12.
Artigo em Inglês | MEDLINE | ID: mdl-29440249

RESUMO

Photobacterium damselae subsp. damselae causes vibriosis in a variety of marine animals, including fish species of importance in aquaculture. It also may cause wound infections in humans that can progress into a fatal outcome. Two major virulence factors are encoded within the large conjugative plasmid pPHDD1: the phospholipase-D damselysin (Dly) and the pore-forming toxin Phobalysin P (PhlyP). The two toxins exert hemolytic and cytolytic activity in a synergistic manner. Albeit PhlyP has close homologues in many Vibrio species, it has unique features that differentiate it from related toxins. Dly phospholipase constitutes a singular trait of P. damselae subsp. damselae among the Vibrionaceae, although related toxins are found in members of the Aeromonadaceae Fish farm outbreaks can also be caused by plasmidless strains. Such observation led to the characterization of two ubiquitous, chromosome-encoded toxins with lesser cytolytic activity: the pore forming-toxin Phobalysin C (PhlyC) and the phospholipase-hemolysin PlpV. Special attention deserves the high genetic diversity of this pathogen, with a number of strain-specific features including the cell envelope polysaccharide synthesis clusters. Fish outbreaks are likely caused by multiclonal populations which contain both plasmidless and pPHDD1-harbouring isolates, and not by well-adapted clonal complexes. Still, among such a genetic heterogeneity, it is feasible to identify conserved weak points in the biology of this bacterium: the two-component regulatory system RstAB (CarSR) was found to be necessary for maximal production of virulence factors and its inactivation severely impaired virulence.

8.
Emerg Infect Dis ; 24(11): 2061-2062, 2018 11.
Artigo em Inglês | MEDLINE | ID: mdl-30334701

RESUMO

In 2015, we detected clinical cases of babesiosis caused by Babesia microti in Yucatán State, Mexico. Cases occurred in 4 children from a small town who became ill during the same month. Diagnosis was confirmed using conventional PCR followed by sequencing of the DNA fragment obtained.


Assuntos
Babesia microti/isolamento & purificação , Babesiose/diagnóstico , Babesiose/parasitologia , Adolescente , Babesia microti/genética , Criança , DNA de Protozoário/química , DNA de Protozoário/genética , Feminino , Humanos , Masculino , México , Reação em Cadeia da Polimerase
9.
Behav Med ; 44(2): 131-140, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-28281936

RESUMO

The association of post-traumatic stress disorder (PTSD) symptom clusters with combat and other operational experiences among United Kingdom Armed Forces (UK AF) personnel who deployed to Afghanistan in 2009 were examined. Previous studies suggest that the risk of developing PTSD rises as combat exposure levels increase. To date, no UK research has investigated how specific classes of combat and operational experiences relate to PTSD symptom clusters. The current study was a secondary analysis of data derived from a two-arm cluster, randomized-controlled trial of a postdeployment operational stress-reduction intervention in deployed UK AF personnel. 2510 UK AF personnel provided combat exposure data and completed the PTSD checklist (civilian version) immediately post-deployment while 1635 of the original cohort completed further followed-up measures four to six months later. A 14-item combat experience scale was explored using principle component analysis, which yielded three main categories of experience: (1) violent combat, (2) proximity to wounding or death and (3) encountering explosive devices. The association of combat experience classes to PTSD 5-factor "dysphoric arousal" model (re-experiencing, avoidance, numbing, dysphoric-arousal and anxious-arousal symptoms) was assessed. Greater exposure to violent combat was predictive of re-experiencing and numbing symptoms, while proximity to wounding or death experiences were predictive of re-experiencing and anxious-arousal symptoms. Explosive device exposure was predictive of anxious-arousal symptoms. The present study suggests that categories of combat experience differentially impact on PTSD symptom clusters and may have relevance for clinicians treating military personnel following deployment.


Assuntos
Militares/psicologia , Transtornos de Estresse Pós-Traumáticos/diagnóstico , Guerra/psicologia , Adolescente , Adulto , Campanha Afegã de 2001- , Feminino , Humanos , Masculino , Modelos Psicológicos , Análise de Componente Principal , Ensaios Clínicos Controlados Aleatórios como Assunto/estatística & dados numéricos , Fatores de Risco , Transtornos de Estresse Pós-Traumáticos/psicologia , Síndrome , Reino Unido , Adulto Jovem
10.
Appl Environ Microbiol ; 83(11)2017 06 01.
Artigo em Inglês | MEDLINE | ID: mdl-28341681

RESUMO

Photobacterium damselae subsp. damselae is a pathogen of marine animals, including fish of importance in aquaculture. The virulence plasmid pPHDD1, characteristic of highly hemolytic isolates, encodes the hemolysins damselysin (Dly) and phobalysin (PhlyP). Strains lacking pPHDD1 constitute the vast majority of the isolates from fish outbreaks, but genetic studies to identify virulence factors in plasmidless strains are scarce. Here, we show that the chromosome I-encoded hemolysin PhlyC plays roles in virulence and cell toxicity in pPHDD1-negative isolates of this pathogen. By combining the analyses of whole genomes and of gene deletion mutants, we identified two hitherto uncharacterized chromosomal loci encoding a phospholipase (PlpV) and a collagenase (ColP). PlpV was ubiquitous in the subspecies and exerted hemolytic activity against fish erythrocytes, which was enhanced in the presence of lecithin. ColP was restricted to a fraction of the isolates and was responsible for the collagen-degrading activity in this subspecies. Consistent with the presence of signal peptides in PlpV and ColP sequences, mutants for the type II secretion system (T2SS) genes epsL and pilD exhibited impairments in phospholipase and collagenase activities. Sea bass virulence experiments and cell culture assays demonstrated major contributions of PhlyC and PlpV to virulence and toxicity.IMPORTANCE This study constitutes genetic and genomic analyses of plasmidless strains of an emerging pathogen in marine aquaculture, Photobacterium damselae subsp. damselae To date, studies on the genetic basis of virulence were restricted to the pPHDD1 plasmid-encoded toxins Dly and PhlyP. However, the vast majority of the recent isolates of this pathogen from fish farm outbreaks lack this plasmid. Here we demonstrate that the plasmidless strains produce two hitherto uncharacterized ubiquitous toxins encoded in chromosome I, namely, the hemolysin PhlyC and the phospholipase PlpV. We report the main roles of these two toxins in fish virulence and in cell toxicity. Our results constitute the basis for a better understanding of the virulence of a widespread marine pathogen.


Assuntos
Cromossomos Bacterianos/genética , Colagenases/metabolismo , Doenças dos Peixes/microbiologia , Infecções por Bactérias Gram-Negativas/veterinária , Fosfolipases/metabolismo , Photobacterium/enzimologia , Photobacterium/patogenicidade , Animais , Bass/microbiologia , Cromossomos Bacterianos/metabolismo , Colagenases/genética , Infecções por Bactérias Gram-Negativas/microbiologia , Proteínas Hemolisinas/genética , Proteínas Hemolisinas/metabolismo , Fosfolipases/genética , Photobacterium/genética , Photobacterium/metabolismo , Plasmídeos/genética , Plasmídeos/metabolismo , Virulência
11.
Behav Med ; 43(4): 307-322, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-27100966

RESUMO

There is significant variation in the way individuals react and respond to extreme stress and adversity. While some individuals develop psychiatric conditions such as posttraumatic stress disorder or major depressive disorder, others recover from stressful experiences without displaying significant symptoms of psychological ill-health, demonstrating stress-resilience. To understand why some individuals exhibit characteristics of a resilient profile, the interplay between neurochemical, genetic, and epigenetic processes over time needs to be explained. In this review, we examine the hormones, neuropeptides, neurotransmitters, and neural circuits associated with resilience and vulnerability to stress-related disorders. We debate how this increasing body of knowledge could also be useful in the creation of a stress-resilient profile. Additionally, identification of the underlying neurobiological components related to resilience may offer a contribution to improved approaches toward the prevention and treatment of stress-related disorders.


Assuntos
Adaptação Psicológica/fisiologia , Encéfalo/metabolismo , Resiliência Psicológica , Transtornos de Estresse Pós-Traumáticos/psicologia , Estresse Psicológico/psicologia , Humanos , Transtornos de Estresse Pós-Traumáticos/metabolismo , Estresse Psicológico/metabolismo , Transmissão Sináptica/fisiologia
12.
Rev Med Chil ; 145(9): 1179-1183, 2017 Sep.
Artigo em Espanhol | MEDLINE | ID: mdl-29424406

RESUMO

Dr. Cora Mayers Glehy, was the director of the Nursing School, head of the Department of Health Education of the General Health Office, a founding member of the Pediatric Society and Dr. Alfredo Demaría Andreani, was a prominent student leader, first sanitarian graduated in the University of John Hopkins, director of Health, professor of the Chair of Hygiene and Preventive Medicine. Both were brilliant academics of the Faculty of Medicine of the University of Chile in the early 1930's and great achievements were expected from them by the medical and university community. These hopes and longings were destroyed when they tragically ended their lives on the morning of a Monday, January 12, 1931.


Assuntos
Docentes de Medicina/história , História da Medicina , Faculdades de Medicina/história , Chile , História do Século XX
13.
Appl Environ Microbiol ; 82(13): 3736-3745, 2016 07 01.
Artigo em Inglês | MEDLINE | ID: mdl-27084008

RESUMO

UNLABELLED: Photobacterium damselae subsp. damselae is considered to be an emerging pathogen of marine fish of importance in aquaculture, with a notable increase in its geographical distribution during the last several years. In this study, we carried out for the first time to our knowledge a genetic and pathobiological characterization of 14 strains isolated from sea bass (Dicentrarchus labrax) reared in the Southeastern Black Sea, where high mortalities were observed at two aquaculture farms during the summer and autumn of 2011. Heterogeneity was evidenced among strains in phenotypical traits, such as sucrose fermentation, motility, and hemolysis. Although 11 of 14 isolates were hemolytic, we found that all of the isolates lacked the pPHDD1 virulence plasmid that encodes the phospholipase-D damselysin (Dly) and the pore-forming toxin PhlyP, two hemolysins previously reported to constitute major virulence factors for turbot. Subsequent PCR and sequencing analyses demonstrated that the 11 hemolytic isolates harbored a complete hlyAch gene, a chromosome I-borne gene that encodes HlyAch hemolysin, whereas the three nonhemolytic isolates contained hlyAch pseudogenes caused by insertion sequence elements. Virulence challenges with two representative strains revealed that, albeit less virulent than the pPHDD1-harboring strain RM-71, the plasmidless hlyAch-positive and hlyAch-negative Black Sea isolates were pathogenic for sea bass. A phylogenetic analysis based on the toxR gene sequence uncovered a greater diversity in the isolates, indicating that the presence of this pathogen in the Black Sea was not caused by the introduction and spread of a single virulent clone but by the proliferation of different clones. IMPORTANCE: The geographical distribution of marine bacterial pathogens is undergoing a worldwide increase. In particular, bacteria of the group vibrios are increasingly being isolated as the causative agents of disease in novel species of cultivated fish in areas where they had not been previously reported. Here we characterize for the first time to our knowledge a collection of isolates of the fish and human pathogen Photobacterium damselae subsp. damselae from diseased sea bass reared in the Black Sea. We uncovered great genetic diversity in the Black Sea isolates of this pathogen, suggesting a multiclonal origin. We also demonstrate for the first time that these isolates bear pathogenic potential for sea bass cultures by virulence challenges.


Assuntos
Doenças dos Peixes/epidemiologia , Doenças dos Peixes/microbiologia , Variação Genética , Genótipo , Infecções por Bactérias Gram-Negativas/veterinária , Photobacterium/classificação , Photobacterium/genética , Animais , Aquicultura , Técnicas de Tipagem Bacteriana , Bass , Mar Negro , Genes Bacterianos , Infecções por Bactérias Gram-Negativas/epidemiologia , Infecções por Bactérias Gram-Negativas/microbiologia , Photobacterium/isolamento & purificação , Photobacterium/fisiologia , Reação em Cadeia da Polimerase , Análise de Sequência de DNA , Fatores de Virulência/genética
14.
Microb Ecol ; 72(4): 851-860, 2016 11.
Artigo em Inglês | MEDLINE | ID: mdl-26552396

RESUMO

The characterization of antibiotic-resistant vibrios isolated from shellfish aquaculture is necessary to elucidate the potential transfer of resistance and to establish effective strategies against vibriosis. With this aim, we analyzed a collection of bacterial isolates obtained from 15 failed hatchery larval cultures that, for the most part, had been treated experimentally with chloramphenicol to prevent vibriosis. Isolates were obtained during a 2-year study from experimental cultures of five different clam species. Among a total of 121 Vibrio isolates studied, 28 were found to be chloramphenicol resistant, suggesting that the shellfish hatchery had been using a sublethal concentration of the antibiotic. Interestingly, chloramphenicol-resistant vibrios showed also resistance to tetracycline and amoxicillin (group A; n = 19) or to streptomycin (group B; n = 9). Chloramphenicol-resistant vibrios were subjected to a PCR amplification and DNA sequencing of the chloramphenicol acetyltransferase genes (cat), and the same approach was followed to study the tetracycline resistance markers (tet). 16S ribosomal RNA (rRNA) gene sequencing revealed that chloramphenicol-resistant vibrios pertained mostly to the Splendidus clade. Conjugation assays demonstrated that various R-plasmids which harbored the cat II/tet(D) genes and cat III gene in groups A and B respectively, were transferred to E. coli and bivalve pathogenic vibrios. Most interestingly, transconjugants exhibited the antibiotic resistance patterns of the donors, despite having been selected only on the basis of chloramphenicol resistance. This is the first report carried out in a bivalve hatchery elucidating the persistence of resistant vibrios, the mechanisms of antibiotic resistance, and the transfer of different R-plasmids.


Assuntos
Antibacterianos/farmacologia , Bivalves/microbiologia , Resistência ao Cloranfenicol/genética , Farmacorresistência Bacteriana Múltipla/genética , Escherichia coli/genética , Pesqueiros , Frutos do Mar/microbiologia , Vibrio/genética , Amoxicilina/farmacologia , Animais , Sequência de Bases , Cloranfenicol/farmacologia , Cloranfenicol O-Acetiltransferase/genética , DNA Bacteriano/genética , Escherichia coli/efeitos dos fármacos , Testes de Sensibilidade Microbiana , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Estreptomicina/farmacologia , Tetraciclina/farmacologia , Vibrio/efeitos dos fármacos , Vibrio/isolamento & purificação
15.
Antonie Van Leeuwenhoek ; 109(8): 1141-52, 2016 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-27230650

RESUMO

Integrating conjugative elements (ICEs) of the SXT/R391 family have been identified in fish-isolated bacterial strains collected from marine aquaculture environments of the northwestern Iberian Peninsula. Here we analysed the variable regions of two ICEs, one preliminarily characterised in a previous study (ICEVscSpa3) and one newly identified (ICEPspSpa1). Bacterial strains harboring these ICEs were phylogenetically assigned to Vibrio scophthalmi and Pseudoalteromonas sp., thus constituting the first evidence of SXT/R391-like ICEs in the genus Pseudoalteromonas to date. Variable DNA regions, which confer element-specific properties to ICEs of this family, were characterised. Interestingly, the two ICEs contained 29 genes not found in variable DNA insertions of previously described ICEs. Most notably, variable gene content for ICEVscSpa3 showed similarity to genes potentially involved in housekeeping functions of replication, nucleotide metabolism and transcription. For these genes, closest homologues were found clustered in the genome of Pseudomonas psychrotolerans L19, suggesting a transfer as a block to ICEVscSpa3. Genes encoding antibiotic resistance, restriction modification systems and toxin/antitoxin systems were absent from hotspots of ICEVscSpa3. In contrast, the variable gene content of ICEPspSpa1 included genes involved in restriction/modification functions in two different hotspots and genes related to ICE maintenance. The present study unveils a relatively large number of novel genes in SXT/R391-ICEs, and demonstrates the major role of ICE elements as contributors to horizontal gene transfer.


Assuntos
Conjugação Genética , Peixes/microbiologia , Pseudoalteromonas/genética , Vibrio/genética , Animais , Aquicultura , Proteínas de Bactérias/genética , Sequência de Bases , Replicação do DNA , Elementos de DNA Transponíveis , DNA Bacteriano/genética , Transferência Genética Horizontal , Genes Bacterianos , Genoma Bacteriano , Filogenia , Análise de Sequência de DNA
16.
Infect Immun ; 83(4): 1246-56, 2015 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-25583529

RESUMO

Photobacterium damselae subsp. damselae is a marine bacterium that causes septicemia in marine animals and in humans. Previously, we had determined a major role of pPHDD1 plasmid-encoded Dly (damselysin) and HlyA (HlyApl) and the chromosome-encoded HlyA (HlyAch) hemolysins in virulence. However, the mechanisms by which these toxins are secreted remain unknown. In this study, we found that a mini-Tn10 transposon mutant in a plasmidless strain showing an impaired hemolytic phenotype contained an insertion in epsL, a component of a type II secretion system (T2SS). Reconstruction of the mutant by allelic exchange confirmed the specific involvement of epsL in HlyAch secretion. In addition, mutation of epsL in a pPHDD1-harboring strain caused an almost complete abolition of hemolytic activity against sheep erythrocytes, indicating that epsL plays a major role in secretion of the plasmid-encoded HlyApl and Dly. This was further demonstrated by analysis of different combinations of hemolysin gene mutants and by strain-strain complementation assays. We also found that mutation of the putative prepilin peptidase gene pilD severely affected hemolysis, which dropped at levels inferior to those of epsL mutants. Promoter expression analyses suggested that impairment of hemolysin secretion in epsL and pilD mutants might constitute a signal that affects hemolysin and T2SS gene expression at the transcriptional level. In addition, single epsL and pilD mutations caused a drastic decrease in virulence for mice, demonstrating a major role of T2SS and pilD in P. damselae subsp. damselae virulence.


Assuntos
Sistemas de Secreção Bacterianos , Proteínas Hemolisinas/metabolismo , Photobacterium/patogenicidade , Fatores de Virulência/metabolismo , Animais , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Sequência de Bases , Endopeptidases/genética , Endopeptidases/metabolismo , Eritrócitos/patologia , Proteínas Hemolisinas/genética , Hemólise , Camundongos , Camundongos Endogâmicos BALB C , Mutação , Photobacterium/genética , Photobacterium/metabolismo , Plasmídeos/genética , Plasmídeos/metabolismo , Análise de Sequência de DNA , Transcrição Gênica , Transposases/genética , Fatores de Virulência/genética
17.
Infect Immun ; 83(11): 4335-48, 2015 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-26303391

RESUMO

Photobacterium damselae subsp. damselae, an important pathogen of marine animals, may also cause septicemia or hyperaggressive necrotizing fasciitis in humans. We previously showed that hemolysin genes are critical for virulence of this organism in mice and fish. In the present study, we characterized the hlyA gene product, a putative small ß-pore-forming toxin, and termed it phobalysin P (PhlyP), for "photobacterial lysin encoded on a plasmid." PhlyP formed stable oligomers and small membrane pores, causing efflux of K(+), with no significant leakage of lactate dehydrogenase but entry of vital dyes. The latter feature distinguished PhlyP from the related Vibrio cholerae cytolysin. Attack by PhlyP provoked a loss of cellular ATP, attenuated translation, and caused profound morphological changes in epithelial cells. In coculture experiments with epithelial cells, Photobacterium damselae subsp. damselae led to rapid hemolysin-dependent membrane permeabilization. Unexpectedly, hemolysins also promoted the association of P. damselae subsp. damselae with epithelial cells. The collective observations of this study suggest that membrane-damaging toxins commonly enhance bacterial adherence.


Assuntos
Toxinas Bacterianas/metabolismo , Proteínas Hemolisinas/metabolismo , Photobacterium/metabolismo , Sequência de Aminoácidos , Animais , Aderência Bacteriana , Toxinas Bacterianas/química , Toxinas Bacterianas/genética , Toxinas Bacterianas/toxicidade , Células Epiteliais/microbiologia , Eritrócitos/citologia , Eritrócitos/efeitos dos fármacos , Proteínas Hemolisinas/química , Proteínas Hemolisinas/genética , Proteínas Hemolisinas/toxicidade , Hemólise , Humanos , Dados de Sequência Molecular , Photobacterium/química , Photobacterium/genética , Coelhos , Alinhamento de Sequência
18.
J Antimicrob Chemother ; 70(3): 914-21, 2015 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-25480492

RESUMO

OBJECTIVES: To determine the association between ertapenem and resistance of Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae and Acinetobacter baumannii-calcoaceticus complex to different antimicrobials while adjusting for relevant hospital factors. METHODS: This was a retrospective time-series study conducted at a tertiary care centre from September 2002 to August 2008. The specific impact of ertapenem on the resistance of these Gram-negative bacilli (GNB) was assessed by multiple linear regression analysis, adjusting for the average length of stay, rate of hospital-acquired infections and use of 10 other antimicrobials, including type 2 carbapenems. RESULTS: Unadjusted analyses revealed significant increases over the duration of the study in the number of GNB resistant to meropenem/imipenem among 1000 isolates each of E. coli (0.46 ±â€Š0.22, P < 0.05), P. aeruginosa (6.26 ±â€Š2.26, P < 0.05), K. pneumoniae (8.06 ±â€Š1.50, P < 0.0005) and A. baumannii-calcoaceticus complex (25.39 ±â€Š6.81, P < 0.0005). Increased resistance to cefepime (and other extended-spectrum cephalosporins) was observed in E. coli (9.55 ±â€Š1.45, P < 0.0005) and K. pneumoniae (15.21 ±â€Š2.42, P < 0.0005). A. baumannii-calcoaceticus complex showed increased resistance to all antimicrobials except amikacin. After controlling for confounders, ertapenem was not significantly associated (P > 0.05) with changes in resistance for any pathogen/antimicrobial combination. CONCLUSIONS: After controlling for confounders, ertapenem was not associated with changes in resistance in a group of sentinel GNB, although significant variations in resistance to different antimicrobials were observed in the unadjusted analyses. These results emphasize the importance of implementation of local resistance surveillance platforms and stewardship programmes to combat the global emergence and spread of antimicrobial resistance.


Assuntos
Antibacterianos/uso terapêutico , Farmacorresistência Bacteriana , Bactérias Gram-Negativas/efeitos dos fármacos , Infecções por Bactérias Gram-Negativas/microbiologia , beta-Lactamas/uso terapêutico , Ertapenem , Infecções por Bactérias Gram-Negativas/tratamento farmacológico , Humanos , Testes de Sensibilidade Microbiana , Estudos Retrospectivos , Vigilância de Evento Sentinela , Centros de Atenção Terciária
19.
Appl Environ Microbiol ; 81(17): 5867-79, 2015 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-26092457

RESUMO

The fish pathogen Photobacterium damselae subsp. piscicida produces the siderophore piscibactin. A gene cluster that resembles the Yersinia high-pathogenicity island (HPI) encodes piscibactin biosynthesis. Here, we report that this HPI-like cluster is part of a hitherto-uncharacterized 68-kb plasmid dubbed pPHDP70. This plasmid lacks homologs of genes that mediate conjugation, but we found that it could be transferred at low frequencies from P. damselae subsp. piscicida to a mollusk pathogenic Vibrio alginolyticus strain and to other Gram-negative bacteria, likely dependent on the conjugative functions of the coresident plasmid pPHDP60. Following its conjugative transfer, pPHDP70 restored the capacity of a vibrioferrin mutant of V. alginolyticus to grow under low-iron conditions, and piscibactin became detectable in its supernatant. Thus, pPHDP70 appears to harbor all the genes required for piscibactin biosynthesis and transport. P. damselae subsp. piscicida strains cured of pPHDP70 no longer produced piscibactin, had impaired growth under iron-limited conditions, and exhibited markedly decreased virulence in fish. Collectively, our findings highlight the importance of pPHDP70, with its capacity for piscibactin-mediated iron acquisition, in the virulence of P. damselae subsp. piscicida. Horizontal transmission of this plasmid-borne piscibactin synthesis gene cluster in the marine environment may facilitate the emergence of new pathogens.


Assuntos
Doenças dos Peixes/microbiologia , Transferência Genética Horizontal , Ilhas Genômicas , Infecções por Bactérias Gram-Negativas/veterinária , Photobacterium/genética , Photobacterium/metabolismo , Plasmídeos/genética , Sideróforos/biossíntese , Animais , Infecções por Bactérias Gram-Negativas/microbiologia , Ferro/metabolismo , Dados de Sequência Molecular , Photobacterium/patogenicidade , Plasmídeos/metabolismo , Virulência
20.
Rev Chilena Infectol ; 32(4): 447-52, 2015 Aug.
Artigo em Espanhol | MEDLINE | ID: mdl-26436791

RESUMO

The first Chair of Microbiology in Chile was created in the School of Medicine of the Cañadilla at the University of Chile in 1892. Dr. Alejandro del Río Soto Aguilar was its first Professor. For almost three decades it was the only educational center for microbiologists in Chile. Among them were the first Professors of the new School of Medicine of the Catholic University of Chile and of the University of Concepción.


Assuntos
Microbiologia/história , Escolas para Profissionais de Saúde/história , Chile , História do Século XIX , História do Século XX , Humanos , Microbiologia/educação
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA