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1.
J Biol Chem ; 299(9): 105119, 2023 09.
Artículo en Inglés | MEDLINE | ID: mdl-37527778

RESUMEN

Serratia marcescens is an opportunistic human pathogen involved in antibiotic-resistant hospital acquired infections. Upon contact with the host epithelial cell and prior to internalization, Serratia induces an early autophagic response that is entirely dependent on the ShlA toxin. Once Serratia invades the eukaryotic cell and multiples inside an intracellular vacuole, ShlA expression also promotes an exocytic event that allows bacterial egress from the host cell without compromising its integrity. Several toxins, including ShlA, were shown to induce ATP efflux from eukaryotic cells. Here, we demonstrate that ShlA triggered a nonlytic release of ATP from Chinese hamster ovary (CHO) cells. Enzymatic removal of accumulated extracellular ATP (eATP) or pharmacological blockage of the eATP-P2Y2 purinergic receptor inhibited the ShlA-promoted autophagic response in CHO cells. Despite the intrinsic ecto-ATPase activity of CHO cells, the effective concentration and kinetic profile of eATP was consistent with the established affinity of the P2Y2 receptor and the known kinetics of autophagy induction. Moreover, eATP removal or P2Y2 receptor inhibition also suppressed the ShlA-induced exocytic expulsion of the bacteria from the host cell. Blocking α5ß1 integrin highly inhibited ShlA-dependent autophagy, a result consistent with α5ß1 transactivation by the P2Y2 receptor. In sum, eATP operates as the key signaling molecule that allows the eukaryotic cell to detect the challenge imposed by the contact with the ShlA toxin. Stimulation of P2Y2-dependent pathways evokes the activation of a defensive response to counteract cell damage and promotes the nonlytic clearance of the pathogen from the infected cell.


Asunto(s)
Autofagia , Interacciones Huésped-Patógeno , Integrina alfa5beta1 , Receptores Purinérgicos P2Y2 , Serratia , Toxinas Biológicas , Animales , Cricetinae , Adenosina Trifosfato/metabolismo , Autofagia/efectos de los fármacos , Células CHO , Cricetulus , Exocitosis/efectos de los fármacos , Interacciones Huésped-Patógeno/efectos de los fármacos , Integrina alfa5beta1/antagonistas & inhibidores , Integrina alfa5beta1/metabolismo , Receptores Purinérgicos P2Y2/metabolismo , Serratia/química , Serratia/efectos de los fármacos , Serratia/fisiología , Toxinas Biológicas/farmacología , Humanos
2.
Acta Biochim Pol ; 68(3): 437-448, 2021 Aug 24.
Artículo en Inglés | MEDLINE | ID: mdl-34428264

RESUMEN

Despite the fact that cephalosporins are rarely used in medical or veterinary treatment, the presence of Enterobacterales strains resistant to this group of anti-bacterial drugs (ESBL) is an important issue that requires attention. Between 2019 and 2021, 14 retention reservoirs, 12 streams, 3 rivers and 1 lake situated in the Tricity area (in northern Poland) were sampled for the presence of ESBL strains. Out of 40 water samples, characteristic growth (E. coli and the KESC group) on Chromagar ESBL plates was observed for 33 samples. The average number of ESBL E. coli was 42±132 CFU/100 ml, while the KESC group was 73±147 CFU/100 ml. Out of 33 positive samples, 57 ESBL Enterobacterales strains were isolated, of which the most abundant species were E. coli (13 isolates) and S. fonticola (23 isolates). The E. coli ESBL isolates not only showed resistance to third generation cephalosporins but also to antibiotics from other groups, such as fluoroquinolones, aminoglycosides and sulfonamides. The S. fonticola ESBL isolates were also found to be mainly resistant to the third generation cephalosporins, with the exception of 5 imipenem and 2 ertapenem-resistant strains. These strains presented highly diverse fingerprinting profiles, as well as significant differences in phenotypic traits helpful for survival in the environment, such as biofilm formation and motility. Moreover, biofilm formation and the swimming ability were species and temperature dependent. We confirmed the presence of highly diverse ESBL strains with multiple drug resistance patterns in the Tricity water reservoirs. This could possibly pose a threat to human health and create a suitable ground for acquiring antibiotics resistance in the natural environment.


Asunto(s)
Antibacterianos/farmacología , Farmacorresistencia Bacteriana Múltiple/efectos de los fármacos , Escherichia coli/efectos de los fármacos , Serratia/efectos de los fármacos , Microbiología del Agua , Biopelículas , Cefalosporinas/farmacología , Microbiología Ambiental , Humanos , Pruebas de Sensibilidad Microbiana , Polonia , Ríos/microbiología , Abastecimiento de Agua/métodos , beta-Lactamasas/metabolismo
3.
Insect Biochem Mol Biol ; 136: 103620, 2021 09.
Artículo en Inglés | MEDLINE | ID: mdl-34216781

RESUMEN

Fleas (Order Siphonaptera) transmit numerous bacterial pathogens that cause severe human diseases (e.g., cat scratch disease, flea-borne spotted fever, murine typhus, plague). Because initial entry of these infectious agents occurs while blood feeding, the immune response in the flea gut is considered to be the first line of defense against invading microbes. However, relatively few studies have identified the flea immune molecules that effectively resist or limit infection in the gut. In other hematophagous insects, an immediate immune response to imbibed pathogens is the generation of reactive oxygen species (ROS). In this study, we utilized cat fleas (Ctenocephalides felis) to investigate whether oral infection with a well-known insect bacterial pathogen (Serratia marcescens) induces ROS synthesis in the flea gut, and whether production of ROS provides a defense mechanism against microbial colonization. Specifically, we treated fleas with an antioxidant to limit the number of free radicals in the digestive tract prior to infection, and then measured the following: S. marcescens infection loads, hydrogen peroxide (ROS) levels, and mRNA abundance of ROS signaling pathway genes. Overall, our data shows that ROS levels increase in response to infection in the flea gut, and that this increase helps to strengthen the flea immune response through the microbicidal activity of ROS.


Asunto(s)
Infecciones Bacterianas/inmunología , Ctenocephalides , Especies Reactivas de Oxígeno/inmunología , Animales , Antioxidantes/farmacología , Ctenocephalides/inmunología , Ctenocephalides/metabolismo , Ctenocephalides/microbiología , Insectos Vectores/inmunología , Insectos Vectores/metabolismo , Insectos Vectores/microbiología , Intestinos/microbiología , Serratia/efectos de los fármacos , Serratia/inmunología
4.
Lett Appl Microbiol ; 72(6): 783-790, 2021 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-33735446

RESUMEN

This study was conducted to isolate and identify extended-spectrum beta-lactamase (ESBL)-producing Enterobacterales in conventional and organic chicken meats, which were sold in Turkey. A total of 200 raw chicken meat sample (100 conventional and 100 organic) were used as material. Classic culture technique based on chromogenic method was used for the isolation of bacteria, and the identification was performed with VITEK MS. Phenotypic ESBL production was detected by combined disc diffusion method. Gene regions responsible for ESBL production were determined by PCR. MIC values of isolates were detected by VITEK 2. Phenotypic ESBL-producing Enterobacterales were detected in 46% of conventional chicken meats and in 22% of organic chicken meats. Of the 115 isolates obtained, 97 (84%) were Escherichia coli, 12 (10%) were Klebsiella pneumoniae, four (3·48%) were Serratia fonticola, one (0·87%) was Rahnella aquatilis, and one (0·87%) was Serratia liquefaciens. PCR analysis revealed that 109 of 115 isolates (94·78%) contained at least one of the blaCTX-M , blaTEM , and blaSHV genes. Of the 115 ESBL-producing isolates, 103 (89·57%) were found resistant to at least one antibiotic except for the ß-lactam group. The contamination level of ESBL-producing Enterobacterales was higher in conventional chicken meats (P < 0·001).


Asunto(s)
Escherichia coli/genética , Klebsiella pneumoniae/genética , Carne/microbiología , Serratia/genética , beta-Lactamasas/genética , Animales , Antibacterianos/farmacología , Pollos/microbiología , Escherichia coli/efectos de los fármacos , Infecciones por Escherichia coli/microbiología , Microbiología de Alimentos , Klebsiella pneumoniae/efectos de los fármacos , Reacción en Cadena de la Polimerasa , Serratia/efectos de los fármacos , Turquía
5.
Pediatr Infect Dis J ; 40(5): 440-445, 2021 05 01.
Artículo en Inglés | MEDLINE | ID: mdl-33264210

RESUMEN

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.


Asunto(s)
Antibacterianos/uso terapéutico , Programas de Optimización del Uso de los Antimicrobianos/métodos , Proteínas Bacterianas/efectos de los fármacos , Resistencia a las Cefalosporinas , Cefalosporinas/uso terapéutico , Infecciones por Enterobacteriaceae/tratamiento farmacológico , beta-Lactamasas/efectos de los fármacos , Adolescente , Niño , Preescolar , Citrobacter/efectos de los fármacos , Estudios de Cohortes , Enterobacter/efectos de los fármacos , Femenino , Humanos , Lactante , Masculino , Morganella/efectos de los fármacos , Estudios Retrospectivos , Factores de Riesgo , Serratia/efectos de los fármacos
6.
Microb Drug Resist ; 27(5): 585-589, 2021 May.
Artículo en Inglés | MEDLINE | ID: mdl-32991273

RESUMEN

Serratia fonticola is a human pathogen widely found in the environment, with birds being reported as possible natural hosts. During an epidemiological and genomic surveillance study conducted to monitor the occurrence of extended-spectrum ß-lactamase (ESBL)-producing Enterobacterales in South American wild birds, we identified an ESBL-positive S. fonticola in a fecal sample collected from a Hudsonian Whimbrel, during its non-breeding range on the Pacific Coast of Chile. Whole genome sequencing analysis and "in silico" modeling revealed a novel variant of the class A ESBLs FONA family, designated FONA-7, which shows 96.28% amino acid identity with FONA-6; with amino acid substitutions occurring in the signal peptide sequence (Thr22→Ser), and in the mature protein (Ser39→Asn and Thr227→Ile). This finding denotes that migratory birds can be potential vectors for the transboundary spread of ESBL-producing bacteria, creating a further theoretical risk for the origin of novel plasmid-encoded ß-lactamases.


Asunto(s)
Antibacterianos/farmacología , Proteínas Bacterianas/genética , Enfermedades de las Aves/microbiología , Serratia/efectos de los fármacos , beta-Lactamasas/genética , Animales , Aves/microbiología , Chile/epidemiología , Vectores de Enfermedades , Heces , Polimorfismo Genético , Serratia/enzimología , Serratia/genética , Secuenciación Completa del Genoma
7.
Sci Rep ; 10(1): 20348, 2020 11 23.
Artículo en Inglés | MEDLINE | ID: mdl-33230153

RESUMEN

The use of microorganisms that allows the recovery of critical high-tech elements such as gallium (Ga) and indium (In) has been considered an excellent eco-strategy. In this perspective, it is relevant to understand the strategies of Ga and In resistant strains to cope with these critical metals. This study aimed to explore the effect of these metals on two Ga/In resistant strains and to scrutinize the biological processes behind the oxidative stress in response to exposure to these critical metals. Two strains of Serratia fonticola, A3242 and B2A1Ga1, with high resistance to Ga and In, were submitted to metal stress and their protein profiles showed an overexpressed Superoxide Dismutase (SOD) in presence of In. Results of inhibitor-protein native gel incubations identified the overexpressed enzyme as a Fe-SOD. Both strains exhibited a huge increase of oxidative stress when exposed to indium, visible by an extreme high amount of reactive oxygen species (ROS) production. The toxicity induced by indium triggered biological mechanisms of stress control namely, the decrease in reduced glutathione/total glutathione levels and an increase in the SOD activity. The effect of gallium in cells was not so boisterous, visible only by the decrease of reduced glutathione levels. Analysis of the cellular metabolic viability revealed that each strain was affected differently by the critical metals, which could be related to the distinct metal uptakes. Strain A3242 accumulated more Ga and In in comparison to strain B2A1Ga1, and showed lower metabolic activity. Understanding the biological response of the two metal resistant strains of S. fonticola to stress induced by Ga and In will tackle the current gap of information related with bacteria-critical metals interactions.


Asunto(s)
Contaminantes Ambientales/farmacología , Galio/farmacología , Indio/farmacología , Serratia/efectos de los fármacos , Superóxido Dismutasa/metabolismo , Proteínas Bacterianas/química , Proteínas Bacterianas/metabolismo , Electrónica/instrumentación , Contaminantes Ambientales/aislamiento & purificación , Contaminantes Ambientales/metabolismo , Galio/aislamiento & purificación , Galio/metabolismo , Humanos , Indio/aislamiento & purificación , Indio/metabolismo , Pruebas de Sensibilidad Microbiana , Estrés Oxidativo/efectos de los fármacos , Especies Reactivas de Oxígeno/agonistas , Especies Reactivas de Oxígeno/metabolismo , Serratia/crecimiento & desarrollo , Serratia/metabolismo , Superóxido Dismutasa/química
8.
Res Vet Sci ; 132: 481-484, 2020 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-32799172

RESUMEN

The transmission of antimicrobial resistance genes from enteric bacteria from the animal reservoir to indigenous bacteria in meat is a serious concern, as it can contribute to human exposure to antimicrobial resistance genes. The aim of this study was to investigate plasmid-mediated horizontal transfer of antimicrobial resistance genes from Escherichia coli to indigenous environmental bacteria in minced pork stored at 10 and 37 °C. E. coli MG1555 containing a gfp-tagged plasmid carrying tetracycline, kanamycin and streptomycin resistance genes was used as the donor with the indigenous bacteria in minced pork acting as potential recipients. The results demonstrated that enteric members of the pork meat microbiota were able to receive gfp-plasmids from the E. coli donor strain at both 10 and 37 °C. The majority of transconjugants were identified as Serratia spp. through sequencing of their 16S rRNA genes. This indicates that environmental Serratia spp. and other Enterobacteriaceae may play a role as carrier of antimicrobial resistance genes through the meat production chain to the consumer.


Asunto(s)
Antibacterianos/farmacología , Escherichia coli/efectos de los fármacos , Escherichia coli/genética , Carne/microbiología , Serratia/efectos de los fármacos , Serratia/genética , Animales , Farmacorresistencia Bacteriana/genética , Enterobacteriaceae/efectos de los fármacos , Microbiología de Alimentos , Transferencia de Gen Horizontal , Pruebas de Sensibilidad Microbiana/veterinaria , Plásmidos , ARN Ribosómico 16S/genética , Porcinos
9.
FEBS Lett ; 594(19): 3095-3107, 2020 10.
Artículo en Inglés | MEDLINE | ID: mdl-32748449

RESUMEN

Protealysin is a thermolysin-like protease of Serratia proteamaculans capable of specifically cleaving actin, which correlates with the invasive activity of these bacteria. Here, we show that inactivation of the protealysin gene does not inhibit invasion but, in contrast, leads to a twofold increase in the S. proteamaculans invasive activity. By mass spectrometry, we identified the outer membrane protein OmpX as a substrate of protealysin. Recombinant E. coli carrying the OmpX gene truncated by 40 N-terminal residues or both the OmpX and protealysin genes, in contrast to the full-length OmpX, do not increase adhesion of these bacteria, indicating that the 40 N-terminal residues of OmpX are indispensable for S. proteamaculans invasion. Our results show that both protealysin and its substrates can stimulate Serratia invasion.


Asunto(s)
Proteínas de la Membrana Bacteriana Externa/metabolismo , Serratia/metabolismo , Serratia/patogenicidad , 2,2'-Dipiridil/farmacología , Células 3T3 , Animales , Adhesión Bacteriana/efectos de los fármacos , Escherichia coli/metabolismo , Galactosa/farmacología , Glucosa/farmacología , Células HeLa , Humanos , Deficiencias de Hierro , Ratones , Proteínas Recombinantes/farmacología , Serratia/efectos de los fármacos , Especificidad por Sustrato/efectos de los fármacos , Termolisina/metabolismo , Factores de Virulencia/metabolismo
10.
J Biotechnol ; 317: 16-26, 2020 Jun 20.
Artículo en Inglés | MEDLINE | ID: mdl-32348830

RESUMEN

Bacterial pigments are potential substitute of chemical photosensitizer for dye-sensitized solar cell (DSSC) due to its non-toxic property and cost-effective production from microbial fermentation. Serratia nematodiphila YO1 was isolated from waterfall in Malaysia and identified using 16S ribosomal RNA. Characterization of the red pigment produced by the bacteria has confirmed the pigment as prodigiosin. Prodigiosin was produced from the fermentation of the bacteria in the presence of different oil substrates. Palm oil exhibited the best performance of cell growth and equivalent prodigiosin yield compared to olive oil and peanut oil. Prodigiosin produced with palm oil supplementation was 93 mg/l compared to 7.8 mg/l produced without supplementation, which recorded 11.9 times improvement. Specific growth rate of the cells improved 1.4 times when palm oil was supplemented in the medium. The prodigiosin pigment produced showed comparable performance as a DSSC sensitizer by displaying an open circuit voltage of 336.1 mV and a maximum short circuit current of 0.098 mV/cm2. This study stands a novelty in proving that the production of prodigiosin is favorable in the presence of palm oil substrate with high saturated fat content, which has not been studied before. This is also among the first bacterial prodigiosin tested as photosensitizer for DSSC application.


Asunto(s)
Fuentes de Energía Bioeléctrica , Reactores Biológicos/microbiología , Aceite de Palma/farmacología , Prodigiosina , Serratia , Medios de Cultivo/química , Medios de Cultivo/farmacología , Concentración de Iones de Hidrógeno , Procesos Fotoquímicos , Prodigiosina/análisis , Prodigiosina/metabolismo , Serratia/efectos de los fármacos , Serratia/metabolismo
11.
J Infect Public Health ; 13(4): 651-656, 2020 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-31522967

RESUMEN

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.


Asunto(s)
Microbioma Gastrointestinal/efectos de los fármacos , Tracto Gastrointestinal/microbiología , Hospitales de Enseñanza/estadística & datos numéricos , Adolescente , Adulto , Anciano , Antibacterianos/farmacología , Niño , Infección Hospitalaria/tratamiento farmacológico , Infección Hospitalaria/microbiología , Farmacorresistencia Bacteriana Múltiple , Escherichia coli/efectos de los fármacos , Femenino , Humanos , Klebsiella/efectos de los fármacos , Masculino , Pruebas de Sensibilidad Microbiana , Persona de Mediana Edad , Morganella/efectos de los fármacos , Nigeria/epidemiología , Prevalencia , Proteus/efectos de los fármacos , Providencia/efectos de los fármacos , Salmonella/efectos de los fármacos , Serratia/efectos de los fármacos , Infecciones Urinarias/tratamiento farmacológico , Infecciones Urinarias/microbiología , Yersinia/efectos de los fármacos , Adulto Joven
12.
Nat Prod Res ; 34(23): 3394-3398, 2020 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-30689407

RESUMEN

Essential oil (EO) from Eriocephalus africanus L. leaves was evaluated against the growth of some phytopathogenic bacteria including Agrobacerium tumifaciens, Dickeya solani, Erwinia amylovora, Pseudomonas cichorii and Serratia pulmithica using the disc diffusion method and minimum inhibitory concentration (MIC) evaluation. Ten compounds in the EO with dominance of Artemisia ketone (2,5,5-trimethyl-2,6-heptadien-4-one) (77.92%) and ledol (19.92%) were revealed. The antibacterial activity indicated efficacy of essential oil against majority of strains isolated. The most effective action was recorded against D. solani, by 7.5 and 10 µL of oil, with 18.33 mm and 100 µg/mL as zone inhibition and MIC, respectively, whereas the lowest activity was exhibited against P. cichorii (diameter inhibition = 6.66 mm at 10 µL of oil, MIC = 100 µg/mL). The strain S. pulmithica appears to be resistant to the oil when the activity is measured by 10 µL of oil but its growth inhibition was reported with a MIC of 100 µg/mL.


Asunto(s)
Antibacterianos/farmacología , Asteraceae/química , Bacterias/efectos de los fármacos , Aceites Volátiles/farmacología , Bacterias/patogenicidad , Productos Agrícolas/microbiología , Evaluación Preclínica de Medicamentos , Farmacorresistencia Bacteriana/efectos de los fármacos , Pruebas de Sensibilidad Microbiana , Monoterpenos/análisis , Aceites Volátiles/química , Hojas de la Planta/química , Pseudomonas/efectos de los fármacos , Serratia/efectos de los fármacos , Sesquiterpenos/análisis
13.
Ecotoxicol Environ Saf ; 185: 109707, 2019 Dec 15.
Artículo en Inglés | MEDLINE | ID: mdl-31561078

RESUMEN

Effective biodegradation of hydrophobic pollutants, such as 1-chloronaphthalene, is strictly associated with the adaptation of environmental bacteria to their assimilation. This study explores the relation between the modifications of cell properties of bacteria belonging to Pseudomonas and Serratia genera resulting from a 12-month exposure to 1-chloronaphthalene, and their biodegradation efficiency. In the presented study, both bacterial strains exhibited higher (70%) degradation of this compound after exposure compared to unexposed (55%) systems. This adaptation can be associated with increased ratio of polysaccharides in the outer layers of bacterial cells, which was confirmed using infrared spectroscopy analysis. Additionally, the analysis of Raman spectra indicated conformational changes of extracellular carbohydrates from α- to ß-anomeric structure. Moreover, the changes in the cell surface hydrophobicity and cell membrane permeability differed between the strains and the Pseudomonas strain exhibited more significant modifications of these parameters. The results suggest that adaptation strategies of both tested strains are different and involve diverse reconstructions of the cell wall and membranes. The results provide a novel and deep insight into the interactions between environmental bacterial strains and chloroaromatic compounds, which opens new perspectives for applying spectrometric methods in investigation of cell adaptation strategies as a result of long-term contact with toxic pollutants.


Asunto(s)
Aclimatación/efectos de los fármacos , Contaminantes Ambientales/análisis , Naftalenos/análisis , Pseudomonas/efectos de los fármacos , Serratia/efectos de los fármacos , Biodegradación Ambiental , Contaminantes Ambientales/toxicidad , Interacciones Hidrofóbicas e Hidrofílicas , Naftalenos/toxicidad , Polisacáridos Bacterianos/metabolismo , Pseudomonas/metabolismo , Serratia/metabolismo
14.
Biochemistry (Mosc) ; 84(6): 663-671, 2019 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-31238866

RESUMEN

Bacterial internalization into eukaryotic cells is ensured by a sophisticated interplay of bacterial and host cell factors. Being a part of cell environment, opportunistic intracellular bacteria have developed various mechanisms providing their interaction with cell surface receptors (E-cadherin, integrins, epidermal growth factor receptor), activation of components of eukaryotic signaling pathways, and facilitation of bacterial uptake, survival, and intracellular replication. Our previous studies on the mechanisms underlying penetration of the opportunistic bacteria Serratia grimesii into cultured eukaryotic cells have shown that pretreatment of the cells with N-acetylcysteine (NAC) promotes S. grimesii invasion, and this effect correlates with the upregulation of E-cadherin expression. Since NAC has been shown to regulate expression of both Src kinase and ROCK, the aim of this work was to reveal the role of these kinases in S. grimesii invasion. We demonstrated that Y-27632, a specific inhibitor of ROCK, significantly promoted invasion of cultured eukaryotic cells by S. grimesii. On the other hand, invasion of the same cells by S. grimesii was inhibited with the Src kinase inhibitor Src-I1 and siRNA directed against RhoA. The effects of the inhibitors correlated with the corresponding changes in the E-cadherin gene expression, upregulation by the ROCK inhibition and downregulation by the Src kinase inhibition. These results prove the participation of ROCK and Src protein kinases in the invasion of eukaryotic cells by the opportunistic pathogen S. grimesii, as well as suggest that other signaling pathways might be involved in S. grimesii uptake, that are promoted by the ROCK inhibition with Y-27632.


Asunto(s)
Inhibidores de Proteínas Quinasas/farmacología , Serratia/efectos de los fármacos , Quinasas Asociadas a rho/antagonistas & inhibidores , Familia-src Quinasas/antagonistas & inhibidores , Acetilcisteína/farmacología , Línea Celular Tumoral , Humanos
15.
Ecotoxicol Environ Saf ; 174: 197-207, 2019 Jun 15.
Artículo en Inglés | MEDLINE | ID: mdl-30826546

RESUMEN

Cadmium (Cd) is a toxic heavy metal and an abiotic stressor to plants; however, inoculation of endophytic bacteria can raise resistance in plants against Cd, as well as improve plant growth. In the present study, two endophytic bacterial strains were isolated from Solanum nigrum, identified as Serratia sp. IU01 and Enterobacter sp. IU02 by 16S DNA sequencing. Both IU01 and IU02 were tolerant up to 9.0 mM of Cd in culture broth and successive increase in Cd concentration from 0 mM to 9.0 mM, led to an increase in the SOD enzyme activity of the isolates. Both strains were capable of indole-3-acetic acid (IAA) synthesis and phosphate solubilization, detected through gas spectrometry-mass chromatography (GC-MS) and Pikovskaya agar medium respectively. Brassica juncea plants stressed with 0-25 mg/kg Cd showed retardation in all growth attributes, however, inoculation of strain IU01 and IU02 significantly promoted the plant growth attributes as compared to control. Moreover, antioxidant enzymes and metabolites against reactive oxygen species (ROS) including polyphenol oxidase (PPO), peroxidase (POD), catalase (CAT), superoxide dismutase (SOD), alcohol dehydrogenase (ADH), reduced glutathione (GSH), malondialdehyde (MDA), flavonoid and polyphenolic contents were also significantly relieved by inoculation of IU01 and IU02 in plant exposed to different concentration of Cd stress as compared to control plants. Phytohormone production, phosphate solubilization, and/or antioxidative support of IU01 and IU02 might be responsible for growth promotion and Cd resistance in the plant.


Asunto(s)
Antioxidantes/metabolismo , Cadmio/toxicidad , Endófitos/aislamiento & purificación , Enterobacter/aislamiento & purificación , Serratia/aislamiento & purificación , Contaminantes del Suelo/toxicidad , Solanum nigrum/microbiología , Biodegradación Ambiental , Enterobacter/efectos de los fármacos , Planta de la Mostaza/efectos de los fármacos , Planta de la Mostaza/microbiología , Especies Reactivas de Oxígeno , Serratia/efectos de los fármacos
16.
Ecotoxicol Environ Saf ; 173: 373-380, 2019 May 30.
Artículo en Inglés | MEDLINE | ID: mdl-30784801

RESUMEN

Glyphosate-based herbicides (GBHs) are the most widespread commonly used broad-spectrum herbicides that contaminate soils and waters, are toxic to bacteria, plants and animals, and have been classified as 'probably carcinogenic to humans' by the International Agency for Research on Cancer in 2015. Particular soil bacteria and fungi can degrade GBHs, hence, search for new GBH-degrading strains or microbial consortia, effective under specific growth conditions and local environment, seems to be a promising solution for bio-remediation of glyphosate-contaminated environment. Consequently, there is a need for rapid and informative methods to evaluate the GBH-induced changes of the metabolic pathways in cells, that may serve as indicators of GBH-degrading potential. Three new GBH-degrading bacterial strains, Pseudomonas sp., Actinobacteria and Serratia sp. were isolated from sludge of municipal waste water treatment plant (Daugavgriva, Riga, Latvia), agricultural soil and plant tissue, respectively. This study examined the response of these isolates to elevated concentrations of glyphosate (GLP) (100 and 500 mg/L) in GBH Klinik® 360 SL. The GBH-induced shift of metabolic activity in cells of Pseudomonas sp. was shown by tests on EcoPlates™. Fourier transform infrared (FTIR) spectroscopy analyses were used to evaluate the metabolomic response of bacteria to elevated concentrations of GBH in the growth environment. The spectra of Pseudomonas sp. and Serratia sp., incubated with and without GBH, were similar, thus indicating their GBH-resistance. The absorption at 1736 cm-1, assigned to ester carbonyl stretch vibrations, was detected in spectra of all three bacteria. The highest ester content was detected in Actinobacteria grown in medium with 1.0% molasses and 100 or 500 mg/L GLP in GBH Klinik®. An increase of cellular amounts of esters, either those of phospholipids or poly-ß-hydroxybutyrates, indicates degradation of GLP. Therefore, monitoring the ester carbonyl stretch vibration band in FTIR spectra of bacterial biomass may speed up the search GBH-degrading strains. Microbiological tests and cell metabolic response studies by FTIR spectroscopy showed that the three new isolates of Pseudomonas sp., Actinobacteria and Serratia sp. were resistant to elevated concentrations of GBH Klinik® in growth environment and exhibited the potential for GBH degradation.


Asunto(s)
Actinobacteria/efectos de los fármacos , Glicina/análogos & derivados , Herbicidas/toxicidad , Pseudomonas/efectos de los fármacos , Serratia/efectos de los fármacos , Actinobacteria/metabolismo , Glicina/toxicidad , Pseudomonas/metabolismo , Serratia/metabolismo , Glifosato
17.
Nucleic Acids Res ; 46(21): 11229-11238, 2018 11 30.
Artículo en Inglés | MEDLINE | ID: mdl-30500953

RESUMEN

The majority of clinically used antibiotics originate from bacteria. As the need for new antibiotics grows, large-scale genome sequencing and mining approaches are being used to identify novel antibiotics. However, this task is hampered by the fact that many antibiotic biosynthetic clusters are not expressed under laboratory conditions. One strategy to overcome this limitation is the identification of signals that activate the expression of silent biosynthetic pathways. Here, we report the use of high-throughput screening to identify signals that control the biosynthesis of the acetyl-CoA carboxylase inhibitor antibiotic andrimid in the broad-range antibiotic-producing rhizobacterium Serratia plymuthica A153. We reveal that the pathway-specific transcriptional activator AdmX recognizes the auxin indole-3-acetic acid (IAA). IAA binding causes conformational changes in AdmX that result in the inhibition of the expression of the andrimid cluster and the suppression of antibiotic production. We also show that IAA synthesis by pathogenic and beneficial plant-associated bacteria inhibits andrimid production in A153. Because IAA is a signalling molecule that is present across all domains of life, this study highlights the importance of intra- and inter-kingdom signalling in the regulation of antibiotic synthesis. Our discovery unravels, for the first time, an IAA-dependent molecular mechanism for the regulation of antibiotic synthesis.


Asunto(s)
Antibacterianos/biosíntesis , Proteínas Bacterianas/genética , Regulación Bacteriana de la Expresión Génica , Ácidos Indolacéticos/farmacología , Serratia/efectos de los fármacos , Factores de Transcripción/genética , Acetil-CoA Carboxilasa/antagonistas & inhibidores , Arabidopsis/crecimiento & desarrollo , Arabidopsis/microbiología , Proteínas Bacterianas/antagonistas & inhibidores , Proteínas Bacterianas/metabolismo , Clonación Molecular , Inhibidores Enzimáticos/metabolismo , Escherichia coli/genética , Escherichia coli/metabolismo , Expresión Génica , Ensayos Analíticos de Alto Rendimiento , Ácidos Indolacéticos/metabolismo , Raíces de Plantas/crecimiento & desarrollo , Raíces de Plantas/microbiología , Plásmidos/química , Plásmidos/metabolismo , Polienos/metabolismo , Unión Proteica , Pirroles/metabolismo , Pythium/efectos de los fármacos , Pythium/crecimiento & desarrollo , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Serratia/genética , Serratia/metabolismo , Transducción de Señal , Factores de Transcripción/antagonistas & inhibidores , Factores de Transcripción/metabolismo , Activación Transcripcional
18.
Sci Rep ; 8(1): 15823, 2018 10 25.
Artículo en Inglés | MEDLINE | ID: mdl-30361680

RESUMEN

Swarming bacteria are an example of a complex, active biological system, where high cell density and super-diffusive cell mobility confer survival advantages to the group as a whole. Previous studies on the dynamics of the swarm have been limited to easily observable regions at the advancing edge of the swarm where cells are restricted to a plane. In this study, using defocused epifluorescence video imaging, we have tracked the motion of fluorescently labeled individuals within the interior of a densely packed three-dimensional (3D) region of a swarm. Our analysis reveals a novel 3D architecture, where bacteria are constrained by inter-particle interactions, sandwiched between two distinct boundary conditions. We find that secreted biosurfactants keep bacteria away from the swarm-air upper boundary, and added antibiotics at the lower swarm-surface boundary lead to their migration away from this boundary. Formation of the antibiotic-avoidance zone is dependent on a functional chemotaxis signaling system, in the absence of which the swarm loses its high tolerance to the antibiotics.


Asunto(s)
Antibacterianos/farmacología , Quimiotaxis , Imagenología Tridimensional , Serratia/citología , Agar , Quimiotaxis/efectos de los fármacos , Vidrio , Mutación/genética , Serratia/efectos de los fármacos , Tensoactivos/farmacología
19.
Lett Appl Microbiol ; 67(5): 465-475, 2018 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-30028030

RESUMEN

Synthesis of gold nanoparticles (AuNPs) has become a needed domain of applied science. Biological method for synthesis of AuNPs by Trichoderma hamatum SU136 aqueous mycelial extract was achieved. The culture filtrate of the fungus was exposed to three different concentrations of gold chloride. The culture filtrate of the fungus was exposed to three different concentrations of gold chloride (0·25, 0·5 and 1·0 mmol l-1 ). In all cases, the gold ions (Au3+ ) were reduced to Au0 , leading to the formation of stable AuNPs. The AuNPs were identified by UV-visible spectrometry, TEM and FT-IR. The presence of a surface plasmon band around 530 nm indicates AuNPs synthesis. Trichoderma hamatum SU136 synthesized 5-30 nm sized; spherical, pentagonal and hexagonal morphologies of AuNPs by TEM. The existence and binding of proteins with nanoparticles was approved by FT-IR study. Parameters optimization showed the smallest size of AuNPs was obtained with (0·5 mmol l-1 gold chloride, pH 7 at 38°C). Interestingly, AuNPs exhibited antimicrobial activity against four pathogenic bacterial strains in the presence of the standard antibiotic, streptomycin. SIGNIFICANCE AND IMPACT OF THE STUDY: Mycosynthesis of AuNPs by Trichoderma hamatum would provide some useful data for oriented biosynthesis of AuNPs. In addition, the applications of mycosynthesized AuNPs were studied against some pathogenic bacteria. Therefore, the gained results detect that these antimicrobial nanoparticles could be explored as hopeful candidates for a variety of biomedical and pharmaceutical applications. This study should provide a further prudence for the fungal-mediated synthesis of AuNPs.


Asunto(s)
Antibacterianos/farmacología , Bacillus subtilis/efectos de los fármacos , Compuestos de Oro/farmacología , Nanopartículas del Metal/química , Pseudomonas aeruginosa/efectos de los fármacos , Serratia/efectos de los fármacos , Staphylococcus aureus/efectos de los fármacos , Trichoderma/metabolismo , Antibacterianos/química , Reactores Biológicos/microbiología , Sinergismo Farmacológico , Oro/química , Compuestos de Oro/química , Espectroscopía Infrarroja por Transformada de Fourier , Estreptomicina/farmacología , Resonancia por Plasmón de Superficie
20.
Metallomics ; 10(8): 1078-1088, 2018 08 15.
Artículo en Inglés | MEDLINE | ID: mdl-29999065

RESUMEN

It has become increasingly apparent that the environmental microorganisms residing in uranium (U) enriched sites offer the possibility of understanding the biological mechanisms catalyzing the processes important for uranium bioremediation. Here, we present the results of uranium biomineralization over a wide pH range by a metal tolerant Serratia sp. strain OT II 7 isolated from the subsurface soil of a U ore deposit at Domiasiat in India. The Serratia cells actively expressed acid and alkaline phosphatase enzymes which hydrolyzed differential amounts of phosphate from an organophosphate substrate in the presence of uranium between pH 5 to 9. These cells precipitated ∼91% uranium from aqueous solutions supplemented with 1 mM uranyl nitrate at pH 5 within 120 h. More rapid precipitation was observed at pH 7 and 9 wherein the cells removed ∼93-94% of uranium from solutions containing 1 mM uranyl carbonate within 24 h. The aqueous uranyl speciation prevalent under the studied pH conditions influenced the localization of crystalline uranyl phosphate precipitates, which in turn, impacted the cell viability to a great extent. Furthermore, the cells tolerated up to ∼1.6 kGy 60Co gamma radiation and their uranium precipitation abilities at pH 5, 7 and 9 were uncompromised even after exposure to a high dose of ionizing radiation. Overall, this study establishes the ecological adaptation of a natural strain like Serratia in a uranium enriched environment and corroborates its contribution towards uranium immobilization in contaminated subsurfaces through the formation of stable uranyl phosphate minerals over a wide pH range.


Asunto(s)
Ácidos/farmacología , Álcalis/farmacología , Biodegradación Ambiental , Biomineralización , Contaminantes Radiactivos/farmacología , Serratia/metabolismo , Uranio/metabolismo , Precipitación Química , Concentración de Iones de Hidrógeno , Fosfatos/metabolismo , Serratia/efectos de los fármacos , Serratia/efectos de la radiación
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