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1.
Anal Bioanal Chem ; 412(8): 1741-1755, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-32043203

RESUMO

Previously, we demonstrated capture and concentration of Salmonella enterica subspecies enterica ser. Typhimurium using magnetic ionic liquids (MILs), followed by rapid isothermal detection of captured cells via recombinase polymerase amplification (RPA). Here, we report work intended to explore the broader potential of MILs as novel pre-analytical capture reagents in food safety and related applications. Specifically, we evaluated the capacity of the ([P66614+][Ni(hfacac)3-]) ("Ni(II)") MIL to bind a wider range of human pathogens using a panel of Salmonella and Escherichia coli O157:H7 isolates, including a "deep rough" strain of S. Minnesota. We extended this exploration further to include other members of the family Enterobacteriaceae of food safety and clinical or agricultural significance. Both the Ni(II) MIL and the ([P66614+][Dy(hfacac)4-]) ("Dy(III)") MIL were evaluated for their effects on cell viability and structure-function relationships behind observed antimicrobial activities of the Dy(III) MIL were determined. Next, we used flow imaging microscopy (FIM) of Ni(II) MIL dispersions made in model liquid media to examine the impact of increasing ionic complexity on MIL droplet properties as a first step towards understanding the impact of suspension medium properties on MIL dispersion behavior. Finally, we used FIM to examine interactions between the Ni(II) MIL and Serratia marcescens, providing insights into how the MIL may act to capture and concentrate Gram-negative bacteria in aqueous samples, including food suspensions. Together, our results provide further characterization of bacteria-MIL interactions and support the broader utility of the Ni(II) MIL as a cell-friendly capture reagent for sample preparation prior to cultural or molecular analyses. Graphical abstract.


Assuntos
Enterobacteriaceae/metabolismo , Líquidos Iônicos/metabolismo , Magnetismo , Enterobacteriaceae/classificação , Enterobacteriaceae/isolamento & purificação , Indicadores e Reagentes/química , Especificidade da Espécie , Água
2.
J Med Microbiol ; 69(2): 228-232, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31922949

RESUMO

Introduction. Rapid and reliable detection of carbapenemase-producing Enterobacterales (CPE) from surveillance cultures is critical in supporting a good infection control programme. We implemented a new algorithm for CPE detection incorporating the NG Test CARBA 5 in January 2019.Aim. Our goals were to compare turnaround time (TAT), costs and staff requirements between the old and new algorithm, and to evaluate the performance of the CARBA 5 test directly on colonies grown on CARBA Smart agar.Methodology. We analysed and compared the TAT of CPE surveillance cultures processed using the old and new CPE screening algorithm. The total actual reagent costs and staff requirements for the new CPE algorithm were compared with the estimated costs and staff requirements of the old CPE algorithm.Results. Of 197 isolates included in the evaluation of the new algorithm, 64 were positive for carbapenemases by both CARBA 5 and Xpert Carba-R assay. Of the 133 that were negative, two were found to harbour NDM and IMI genotypes. Significant improvements in TAT were achieved with 88.7 % of cultures with CPE, reported on the same day as growth was observed on CARBA Smart agar compared to none in the old algorithm. The new algorithm incurred lower costs and, based on our workload, the new algorithm is estimated to save 28.9 man-hours annually.Conclusion. CARBA 5 performs well on colonies growing on CARBA Smart agar and significant improvements in TAT can be achieved without incurring additional costs or staff requirements.


Assuntos
Proteínas de Bactérias/metabolismo , Técnicas de Laboratório Clínico/métodos , Contagem de Colônia Microbiana/métodos , Infecções por Enterobacteriaceae/microbiologia , Enterobacteriaceae/crescimento & desenvolvimento , Enterobacteriaceae/isolamento & purificação , Ensaios Enzimáticos/métodos , Algoritmos , Proteínas de Bactérias/genética , Técnicas de Laboratório Clínico/economia , Contagem de Colônia Microbiana/economia , Enterobacteriaceae/enzimologia , Enterobacteriaceae/genética , Infecções por Enterobacteriaceae/diagnóstico , Ensaios Enzimáticos/economia , Humanos , Sensibilidade e Especificidade , beta-Lactamases/genética , beta-Lactamases/metabolismo
3.
J Med Microbiol ; 69(2): 233-238, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31971501

RESUMO

Introduction. Raoultella spp. representatives are Gram-negative rod-shaped bacteria of the Enterobacteriaceae family. These bacteria are commonly found in the natural environment.Aim. The aim of the study was to indicate the reliable method for Raoultella spp. strains identification, evaluate the susceptibility of Raoultella spp. strains to selected antimicrobials and to detect their resistance mechanisms to beta-lactams.Methodology. Susceptibility of the strains to chosen antimicrobials was determined using the automatic method. The presence of particular antimicrobial resistant mechanism and genes encoding ESBLs and MBLs was determined respectively with double-disc synergy test and commercially available kit - eazyplex SuperBug CRE test (Amplex Diagnostics) and standard PCR. For the selected strains, DNA sequencing was performed.Results. Amongst 105 of the examined Raoultella spp. strains, majority were sensitive to: imipenem (99.0 %), meropenem (98.1 %), gentamicin (93.3 %) and ciprofloxacin (92.4 %). Of the tested Raoultella strains, thirteen (12.4 %) produced ESBLs and one strain simultaneously ESBLs and MBLs. The DNA sequencing results were as follows: for all the reference strains the correct species identification was achieved, for the analysed strains two were identified as R. planticola and one as R. ornithinolytica.Conclusion. Although Raoultella spp. strains remain sensitive to antibiotics, there is a constant need to monitor the sensitivity of these bacteria to selected antimicrobials. Isolation of a multi-drug resistant R. ornithinolytica strain indicates that even the less frequently isolated species of Enterobacteriaceae family should be precisely identified because they might be of clinical importance and the particular strain can also produce enzymes that pose the greatest threat today.


Assuntos
Antibacterianos/farmacologia , Farmacorresistência Bacteriana , Infecções por Enterobacteriaceae/microbiologia , Enterobacteriaceae/efeitos dos fármacos , Enterobacteriaceae/isolamento & purificação , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Enterobacteriaceae/enzimologia , Enterobacteriaceae/genética , Humanos , Imipenem/farmacologia , Meropeném/farmacologia , Testes de Sensibilidade Microbiana , beta-Lactamases/genética , beta-Lactamases/metabolismo , beta-Lactamas/farmacologia
4.
J Med Microbiol ; 69(2): 207-217, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31976856

RESUMO

Introduction. Infections attributable to carbapenem-resistant Gram-negative bacilli are increasing globally. New antimicrobial agents are urgently needed to treat patients with these infections.Aim. To describe susceptibility to the novel carbapenem-ß-lactamase inhibitor combination imipenem-relebactam and comparators of clinical isolates of non-Proteeae Enterobacteriaceae (NPE) and Pseudomonas aeruginosa from intraabdominal infections (IAIs) and urinary tract infections (UTIs).Methods. Broth microdilution MICs were determined for isolates collected in 22 European countries in 2015-2017 and interpreted using EUCAST breakpoints; imipenem-relebactam MICs were interpreted using imipenem breakpoints.Results. For NPE, 98.4 % of isolates from IAIs (n=10,465) and 98.5 % of UTI isolates (n=7,446) were susceptible to imipenem-relebactam, as were 42.4 % of imipenem-nonsusceptible (n=474), 98.6 % of Klebsiella pneumoniae carbapenemase (KPC)-positive (n=138), and 93.9 % of multidrug-resistant (MDR) isolates (n=4,424) from IAIs and UTIs combined. Molecular analysis demonstrated that two-thirds of imipenem-nonsusceptible isolates rendered susceptible by relebactam carried KPCs; 96 % (261/271) of imipenem-nonsusceptible isolates of NPE that remained nonsusceptible in the presence of relebactam carried metallo-ß-lactamase (MBL)-type and/or OXA-48-like carbapenemases. Among P. aeruginosa, 94.4 % of IAI (n=1,245) and 93.0 % of UTI isolates (n=714) were susceptible to imipenem-relebactam, as were 74.4 % of imipenem-nonsusceptible (n=469) and 79.8 % of MDR isolates (n=595) from IAIs and UTIs combined. Among the 120 isolates of P. aeruginosa that remained nonsusceptible to imipenem upon addition of relebactam, 72 % carried MBLs. The distribution of NPE and P. aeruginosa carrying carbapenemases varied substantially across Europe, as did resistance to imipenem and imipenem-relebactam.Conclusions. Continued surveillance of antimicrobial resistance and resistance mechanisms, including the study of imipenem-relebactam as it approaches regulatory approval, appears warranted.


Assuntos
Antibacterianos/farmacologia , Compostos Azabicíclicos/farmacologia , Farmacorresistência Bacteriana , Enterobacteriaceae/efeitos dos fármacos , Imipenem/farmacologia , Infecções Intra-Abdominais/microbiologia , Pseudomonas aeruginosa/efeitos dos fármacos , Infecções Urinárias/microbiologia , Enterobacteriaceae/isolamento & purificação , Enterobacteriaceae/fisiologia , Infecções por Enterobacteriaceae/epidemiologia , Infecções por Enterobacteriaceae/microbiologia , Monitoramento Epidemiológico , Europa (Continente)/epidemiologia , Humanos , Infecções Intra-Abdominais/epidemiologia , Testes de Sensibilidade Microbiana , Infecções por Pseudomonas/epidemiologia , Infecções por Pseudomonas/microbiologia , Pseudomonas aeruginosa/isolamento & purificação , Pseudomonas aeruginosa/fisiologia , Infecções Urinárias/epidemiologia
5.
BMC Infect Dis ; 20(1): 75, 2020 Jan 23.
Artigo em Inglês | MEDLINE | ID: mdl-31973704

RESUMO

BACKGROUND: Healthcare-Associated Infections (HAIs) represent one of the leading issues to patient safety as well as a significant economic burden. Similarly, Antimicrobial Use (AMU) and Resistance (AMR) represent a growing threat to global public health and the sustainability of healthcare services. METHODS: A Point Prevalence Survey (PPS) following the 2016 ECDC protocol for HAI prevalence and AMU was conducted at Ferrara University Hospital (FUH). Data were collected by a team of trained independent surveyors in 2016 and 2018. Risk factors independently associated with HAI were assessed by a multivariate logistic regression model. RESULTS: Of the 1102 patients surveyed, 115 (10.4%) had an active HAI and 487 (44.2%) were on at least 1 systemic antimicrobial agent. Factors independently associated with increased HAI risk were a "Rapidly Fatal" McCabe score (expected fatal outcome within 1 year), presence of medical devices (PVC, CVC, indwelling urinary catheter or mechanically assisted ventilation) and a length of hospital stay of at least 1 week. The most frequent types of HAI were pneumonia, bloodstream infections, and urinary tract infections. Antimicrobial resistance to third-generation cephalosporins was observed in about 60% of Enterobacteriaceae. CONCLUSIONS: The survey reports a high prevalence of HAI and AMU in FUH. Repeated PPSs are useful to control HAIs and AMU in large acute-care hospitals, highlighting the main problematic factors and allowing planning for improvement actions.


Assuntos
Anti-Infecciosos/uso terapêutico , Bacteriemia/epidemiologia , Infecção Hospitalar/epidemiologia , Enterobacteriaceae/isolamento & purificação , Pneumonia/epidemiologia , Infecções Urinárias/epidemiologia , Adolescente , Adulto , Idoso , Bacteriemia/tratamento farmacológico , Infecção Hospitalar/tratamento farmacológico , Feminino , Hospitais Universitários , Humanos , Itália/epidemiologia , Masculino , Pessoa de Meia-Idade , Pneumonia/tratamento farmacológico , Prevalência , Fatores de Risco , Inquéritos e Questionários , Cateteres Urinários/microbiologia , Infecções Urinárias/tratamento farmacológico , Adulto Jovem
6.
Syst Appl Microbiol ; 43(2): 126052, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31932140

RESUMO

Acute oak decline (AOD) affects native UK oak species causing rapid decline and mortality in as little as five years. A major symptom of AOD is black weeping stem lesions associated with bacterial phytopathogens, Brenneria goodwinii and Gibbsiella quercinecans. However, there is limited knowledge on the ecological and environmental reservoirs of these phytopathogens. Rainwater and soils are common reservoirs of plant pathogens in a forest environment; therefore, the aim of this study was to investigate the survival of B. goodwinii and G. quercinecans in vitro when inoculated into rainwater and forest soil using a combination of agar-based colony counts and gyrB gene-targeted quantitative PCR (qPCR). Brenneria goodwinii lost viability on inoculation into soil and rainwater, but was detectable at low abundance in soil for 28 days using qPCR, suggesting a limited ability to persist outside of the host, potentially in a viable but non-culturable (VBNC) state. Conversely, Gibbsiella quercinecans, was re-isolated from rainwater for the entire duration of the experiment (84 days) and was re-isolated from forest soil after 28 days, with qPCR analysis corroborating these trends. These data demonstrate that B. goodwinii is unable to survive in forest soils and rainwater, suggesting that it may be an endosymbiont of oak trees, whereas G. quercinecans remains viable in soil and rainwater biomes, suggesting a broad ecological distribution. These data advance understanding of the potential epidemiology of AOD-associated bacteria and their ecological reservoirs, thus increasing the overall knowledge of the pathology of AOD, which assists the development of future management strategies.


Assuntos
Enterobacteriaceae/fisiologia , Florestas , Gammaproteobacteria/fisiologia , Doenças das Plantas/microbiologia , Quercus/microbiologia , Chuva/microbiologia , Microbiologia do Solo , Enterobacteriaceae/crescimento & desenvolvimento , Enterobacteriaceae/isolamento & purificação , Gammaproteobacteria/crescimento & desenvolvimento , Gammaproteobacteria/isolamento & purificação , Viabilidade Microbiana , Especificidade da Espécie
7.
Recent Pat Biotechnol ; 14(1): 63-77, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31577211

RESUMO

AIMS: Our exploration work has uncovered the different anti-toxin/metal tolerance and patterns against the heavy metal resistant coliform microscopic organisms from the aquatic waste of the hospital. It might give new routes for the treatment of irresistible ailments particularly by coliform and critical for hazard evaluation as well as hazard management associated with the effluents of the hospital. BACKGROUND: The higher use of pharmaceuticals, Radionuclides, and other antimicrobial solvents are the major source of metals in hospital wastewater. The hospital aquatic environment has a high content of both organic and inorganic matter with living organisms. Bacteria can resist an antimicrobial agent by producing extracellular enzymes that eliminate antibiotics and metal toxicity. In this study, we covered the existing patent literature in this area. New patents in the areas of topically applied antibiotics and agents that can potentiate the achievement of existing antibiotics may extend their helpful lifetime. METHODS: Samples were collected from three different Departments of King George Medical University, Lucknow during the month of December to May (2015-16). Isolation and metal tolerance of coliform isolates were done on metal amended plates. The antibiotic sensitivity test was done by disc diffusion method. The plasmid DNA of bacterial isolates was done by the alkaline lysis method. The conjugation study was also performed in wastewater as well as a nutrient medium. RESULTS: Maximum isolates demonstrated their MICs at 400, 800 and 1600 µg/ml against all the metals, respectively. The high level of resistance was observed against Methicillin (88.32%, 80.60%) followed by penicillin (75%, 76%), Cephradin (59.52%, 28.84%) and least to Gentamycine (1.92%, 5.76) in E. coli and Enterobacter, respectively. Of 70%, 78% E. coli and Enterobacter isolates produce beta-lactamase activity. Six amino acid residues namely, Glu104, Tyr105, Asn132, Asn170, Ala237, and Gly238 of the beta-lactamase were found in the common interaction with the selected drugs. Plasmid DNA size ranged between 48-58.8 kb. The conjugation experiments showed a higher transfer frequency (5.5×10-1 and 3.6×10-1) rate among antibiotics and metals tested. CONCLUSION: The finding of this study presents a potential health problem as the predominant coliform species have increasingly been associated with outbreaks of hospital infections. It is recommended that hospital waste must be properly treated before its release into the environment.


Assuntos
Antibacterianos/farmacologia , Resistência Microbiana a Medicamentos , Enterobacteriaceae , Hospitais , Metais Pesados/farmacologia , Águas Residuárias/microbiologia , Enterobacteriaceae/efeitos dos fármacos , Enterobacteriaceae/enzimologia , Enterobacteriaceae/isolamento & purificação , Índia , Patentes como Assunto , beta-Lactamases
8.
Ann Otol Rhinol Laryngol ; 129(1): 5-11, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31390881

RESUMO

OBJECTIVES: Intranasal septal splints are often used in nasal septal surgeries. Routine use of postoperative antibiotics is an accepted practice, although data regarding its efficacy in preventing postsurgical complications are limited. This study aimed to examine bacterial colonization on septal splints following prophylactic antibiotic therapy and the association with postoperative infections. METHODS: Fifty-five patients underwent septoplasty by a single surgeon between March 2015 and April 2016. All had intranasal septal splints and were given antibiotic prophylaxis for 7 days until removal of splints. Nasal cultures were taken before surgery, and septal splints were examined for bacterial colonization following their removal. RESULTS: Thirty-six patients (65%) had positive nasal culture prior to surgery. The most common isolates were Staphylococcus aureus (30%) and Enterobacteriaceae species (66%). All these patients had postoperative bacterial colonization on septal splints. In 15 patients with negative preoperative cultures, bacteria were isolated postoperatively. An increased resistance profile was documented postoperatively in 9 patients (16%), including two with multidrug resistance. In two of these patients preoperative wild-type strains acquired antibiotic resistance postoperatively. No adverse drug reactions to antibiotics were reported. CONCLUSIONS: Increased bacterial growth and emergence of resistant strains were observed on intranasal septal splints despite prophylactic antibiotic treatment. Nonetheless, this did not translate into clinical infection. Thus, considering antibiotics overuse and increasing bacterial resistance, further research is needed to determine the role of antibiotic prophylaxis in the setting of intranasal splints.


Assuntos
Antibacterianos/uso terapêutico , Antibioticoprofilaxia , Cavidade Nasal/microbiologia , Septo Nasal/cirurgia , Procedimentos Cirúrgicos Nasais/métodos , Contenções/microbiologia , Infecção da Ferida Cirúrgica/prevenção & controle , Adolescente , Adulto , Idoso , Combinação Amoxicilina e Clavulanato de Potássio/uso terapêutico , Cefalexina/uso terapêutico , Técnicas de Cultura , Farmacorresistência Bacteriana , Farmacorresistência Bacteriana Múltipla , Enterobacteriaceae/isolamento & purificação , Infecções por Enterobacteriaceae/epidemiologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Infecções Estafilocócicas/epidemiologia , Staphylococcus aureus/isolamento & purificação , Infecção da Ferida Cirúrgica/epidemiologia , Adulto Jovem
9.
Lett Appl Microbiol ; 70(1): 42-47, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31642085

RESUMO

The increasing frequency of class A KPC enzymes, class B metallo-ß-lactamases (MBLs) and class D OXA-48 enzymes in Enterobacteriaceae makes their early identification urgent. A simple commercial MASTDISCS combi Carba plus disc system (MAST-Carba plus) was designed for the identification of MBLs, KPC and OXA-48 carbapenemase genes in Enterobacteriaceae. To validate the MAST-Carba plus, a total of 77 isolates of carbapenemase-producing Enterobacteriaceae (CPE) and 84 isolates of noncarbapenemase-producing Enterobacteriaceae (non-CPE) were selected for differentiation of the genes of Enterobacteriaceae by MAST-Carba plus. Meanwhile, the carbapenemase genes such as blaKPC , blaIMP , blaVIM , blaNDM-1 and blaOXA-48 were detected by PCR (polymerase chain reaction). Thus, when considered on the basis of PCR results, the sensitivity of MAST-Carba plus detection of KPC strains is 82·3%, the specificity is 100·0%, the positive predictive value is 100·0% and the negative predictive value is 92·4%. For MBLs strains, the sensitivity is 100·0%, the specificity is 97·1%, the positive predictive value is 84·6% and the negative predictive value is 100·0%. For OXA-48 strains, the sensitivity is 100·0%, the specificity is 99·4%, the positive predictive value is 80·0% and the negative predictive value is 100·0%. Our findings suggest that MAST-Carba plus is a rapid and promising method for identifying the MBLs, KPC and OXA-48 carbapenemase genes in Enterobacteriaceae, which could be exploited in basic microbiology laboratory to prevent the transmission of CPE. SIGNIFICANCE AND IMPACT OF THE STUDY: Not only detection of carbapenemases but also identification of their genes accurately and rapidly in Enterobacteriaceae is still a major challenge for clinical laboratories in order to prevent the transmission of carbapenemase-producing Enterobacteriaceae (CPE). Therefore, this study aimed to evaluate the performance of a new rapid method (MASTDISCS combi Carba plus) for the identification of metallo-ß-lactamases (MBLs), KPC and OXA-48 carbapenemase genes in Enterobacteriaceae clinical isolates.


Assuntos
Proteínas de Bactérias/análise , Infecções por Enterobacteriaceae/microbiologia , Enterobacteriaceae/enzimologia , Enterobacteriaceae/isolamento & purificação , Ensaios Enzimáticos/métodos , beta-Lactamases/análise , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Enterobacteriaceae/classificação , Enterobacteriaceae/genética , Humanos , Testes de Sensibilidade Microbiana , Reação em Cadeia da Polimerase , Sensibilidade e Especificidade , beta-Lactamases/genética , beta-Lactamases/metabolismo
10.
J Ethnopharmacol ; 247: 112299, 2020 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-31606537

RESUMO

ETHNOPHARMACOLOGICAL RELEVANCE: Hua-Feng-Dan (HFD) is a traditional Chinese medicine used for neurological disorders. HFD contains cinnabar (HgS) and realgar (As4S4). The ethnopharmacological basis of cinnabar and realgar in HFD is not known. AIM OF THE STUDY: To address the role of cinnabar and realgar in HFD-produced neuroprotection against neurodegenerative diseases and disturbance of gut microbiota. MATERIALS AND METHODS: Lipopolysaccharide (LPS) plus rotenone (ROT)-elicited rat dopaminergic (DA) neuronal damage loss was performed as a Parkinson's disease animal model. Rats were given a single injection of LPS. Four months later, rats were challenged with the threshold dose of ROT. The clinical dose of HFD was administered via feed, starting from ROT administration for 46 days. Behavioral dysfunction was detected by rotarod and Y-maze tests. DA neuron loss and microglial activation were assessed via immunohistochemical staining and western bolt analysis. The colon content was collected to extract bacterial DNA followed by real-time PCR analysis with 16S rRNA primers. RESULTS: LPS plus ROT induced neurotoxicity, as evidenced by DA neuron loss in substantia nigra, impaired behavioral functions and increased microglial activation. HFD-original (containing 10% cinnabar and 10% realgar) rescued loss of DA neurons, improved behavioral dysfunction and attenuated microglial activation. Compared with HFD-original, HFD-reduced (3% cinnabar and 3% realgar) was also effective, but to be a less extent, while HFD-removed (without cinnabar and realgar) was ineffective. In analysis of gut microbiome, the increased Verrucomicrobiaceae and Lactobacteriaceae, and the decreased Enterobacteeriaceae by LPS plus ROT were ameliorated by HFD-original, and to be the less extent by HFD-reduced. CONCLUSION: Cinnabar and realgar are active ingredients in HFD to exert beneficial effects in a neurodegenerative model and gut microbiota.


Assuntos
Arsenicais/farmacologia , Medicamentos de Ervas Chinesas/farmacologia , Microbioma Gastrointestinal/efeitos dos fármacos , Compostos de Mercúrio/farmacologia , Fármacos Neuroprotetores/farmacologia , Síndromes Neurotóxicas/tratamento farmacológico , Sulfetos/farmacologia , Animais , Arsenicais/química , Arsenicais/uso terapêutico , DNA Bacteriano/isolamento & purificação , Modelos Animais de Doenças , Neurônios Dopaminérgicos/efeitos dos fármacos , Neurônios Dopaminérgicos/patologia , Medicamentos de Ervas Chinesas/química , Medicamentos de Ervas Chinesas/uso terapêutico , Enterobacteriaceae/efeitos dos fármacos , Enterobacteriaceae/genética , Enterobacteriaceae/isolamento & purificação , Etnofarmacologia , Microbioma Gastrointestinal/genética , Microbioma Gastrointestinal/imunologia , Humanos , Mediadores da Inflamação/metabolismo , Lactobacillaceae/efeitos dos fármacos , Lactobacillaceae/genética , Lactobacillaceae/isolamento & purificação , Lipopolissacarídeos/toxicidade , Masculino , Compostos de Mercúrio/química , Compostos de Mercúrio/uso terapêutico , Microglia/efeitos dos fármacos , Microglia/imunologia , Microglia/patologia , Degeneração Neural , Fármacos Neuroprotetores/química , Fármacos Neuroprotetores/uso terapêutico , Síndromes Neurotóxicas/etiologia , Síndromes Neurotóxicas/imunologia , Síndromes Neurotóxicas/patologia , RNA Ribossômico 16S/genética , Ratos , Rotenona/toxicidade , Sulfetos/química , Sulfetos/uso terapêutico , Verrucomicrobia/efeitos dos fármacos , Verrucomicrobia/genética , Verrucomicrobia/isolamento & purificação
11.
Meat Sci ; 160: 107967, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31655244

RESUMO

This study investigates the microbiological conditions before maturation of wild boar meat (Sus scrofa) processed in approved game handling establishments in Italy. Fillets and legquarters of 37 carcasses were tested to assess Aerobic Colony Count (ACC), Enterobacteriaceae Count (EC) and Salmonella presence. Salmonella was never found and mean values of ACC and EC were 4.67 ±â€¯1.78 SD and 2.60 ±â€¯1.58 SD log CFU/cm2, respectively. Both ACC and EC increased with time between evisceration and skinning, were significantly higher in fillets and when meat was processed by untrained operators. ACC also increased with boars' weight and when carcasses were cleaned with running potable water. Based on limits set by EU Regulation No 1441/2007 for pork meat, most legquarters resulted satisfactory or acceptable (59% for ACC and 70% for EC), while most fillets were unsatisfactory (76% ACC, 78% EC). Results show that the wild game meat supply chain can be a safe process when handling practices reported in European and National regulations are met.


Assuntos
Manipulação de Alimentos/métodos , /microbiologia , Animais , Bactérias Aeróbias/isolamento & purificação , Peso Corporal , Contagem de Colônia Microbiana/veterinária , Enterobacteriaceae/isolamento & purificação , Microbiologia de Alimentos , Itália , Sus scrofa
12.
World J Microbiol Biotechnol ; 35(11): 172, 2019 Oct 31.
Artigo em Inglês | MEDLINE | ID: mdl-31673798

RESUMO

The present study was designed with the objective of improving growth and nodulation of soybean [Glycine max (L.) Merill] with co-inoculation of native Bradyrhizobium sp. (LSBR-3) (KF906140) and non-rhizobial nodule endophytic diazotroph Leclercia adecarboxylata (LSE-1) (KX925974) with multifunctional plant growth promoting (PGP) traits in cereal based cropping system (Rice-Wheat). A total of 40 endophytic bacteria from cultivated and wild sp. of soybean were screened for multifarious PGP traits and pathogenicity test. Based on PGP traits, antagonistic activities and bio-safety test; L. adecarboxylata (LSE-1) was identified with 16 S rRNA gene sequencing along with the presence of nifH (nitrogen fixation) and ipdc (IAA production) genes. Dual inoculant LSE-1 and LSBR-3 increased indole acetic acid (IAA), P & Zn-solubilization, 1-aminocyclopropane-1-carboxylate deaminase (ACCD) activity, siderophore, biofilm formation and exo-polysaccharides in contrast to single inoculation treatment. Further, assessment of dual inoculant LSBR-3 + LSE-1 improved growth parameters, nodulation, soil enzymes activities, nutrient accumulation and yield as compared to single as well as un-inoculated control treatment under field conditions. Single inoculant LSBR-3 improved yield by 8.84% over control. Further, enhancement of 4.15% grain yield was noticed with LSBR-3 + LSE-1 over LSBR-3 alone treatment. Application of LSBR-3 + LSE-1 gave superior B:C ratio (1.29) and additional income approximately 116 USD ha-1 in contrast to control treatment. The present results thus, is the first report of novel endophytic diazotroph L. adecarboxylata (LSE-1) as PGPR from Indian conditions particularly in Punjab region for exploiting as potential PGPR along with Bradyrhizobium sp. (LSBR-3) in soybean.


Assuntos
Bradyrhizobium/isolamento & purificação , Bradyrhizobium/fisiologia , Enterobacteriaceae/isolamento & purificação , Enterobacteriaceae/fisiologia , Desenvolvimento Vegetal , Raízes de Plantas/microbiologia , Soja/crescimento & desenvolvimento , Soja/microbiologia , Proteínas de Bactérias/genética , Biofilmes/crescimento & desenvolvimento , Bradyrhizobium/classificação , Bradyrhizobium/genética , Endófitos , Enterobacteriaceae/classificação , Enterobacteriaceae/genética , Fertilizantes , Ácidos Indolacéticos/metabolismo , Fixação de Nitrogênio/genética , Nutrientes , Oryza/microbiologia , Oxirredutases/genética , Filogenia , RNA Ribossômico 16S/genética , Nódulos Radiculares de Plantas/microbiologia , Sideróforos , Soja/química , Triticum/microbiologia
13.
Klin Lab Diagn ; 64(10): 613-619, 2019.
Artigo em Russo | MEDLINE | ID: mdl-31742955

RESUMO

Primarily in the study of bacteremia, coagulase-negative staphylococci and representatives of the order Enterobacterales were found. To establish the etiological role of coagulase-negative staphylococcus in making a diagnosis of sepsis in each particular case, it is necessary to consider the condition and age of the child, as well as indicators of markers of systemic inflammation. In 1/3 cases of coagulase-negative staphylococcus indicate colonization of the catheter. Staphylococcus aureus in bacteremia and sepsis - 6.5%, Haemophilus influenzae - 0.6%, Esherichia coli - 7.8%, Streptococcus agalactiae - 2%. For the diagnosis of sepsis, it is necessary to conduct repeated (at least two times) blood culture studies using high-quality nutrient media containing all the necessary growth factors, followed by a mandatory determination of the susceptibility of the isolated strains of microorganisms to antimicrobial agents. Bacteremia as a whole in children of a multidisciplinary hospital amounted to 5.8%. In premature babies, bacteremia was detected in 4.4% of cases, of which sepsis was confirmed in 41,2%.


Assuntos
Bacteriemia/diagnóstico , Sepse/diagnóstico , Criança , Enterobacteriaceae/isolamento & purificação , Hospitais , Humanos , Lactente , Testes de Sensibilidade Microbiana , Infecções Estafilocócicas/diagnóstico , Staphylococcus/isolamento & purificação
14.
Klin Lab Diagn ; 64(10): 627-631, 2019.
Artigo em Russo | MEDLINE | ID: mdl-31742957

RESUMO

Relevance of microbiological monitoring in chronic osteomyelitis of the tibia developed during treatment of fractures with a plate is associated with a noticeable increase of various kinds of the microflora. A microbiological study was conducted of pathological material taken from wounds, fistulas and from the focus of inflammation in 49 patients with chronic tibial osteomyelitis, developed during treatment of fractures with a plate. The patients underwent sequestrectomy of the tibia and subsequent application of bilocal consecutive compression-distraction osteosynthesis or monolocal compression osteosynthesis. Microbiological study of smears taken before the reconstructive treatment from fistulas and wounds of patients showed that in monoculture there were 30 strains, the remaining 30 - as a part of 14 two - and three-component associations. The frequency of S. aureus strains was 55.3%, followed by coagulase-negative staphylococcus - 13.6% and representatives of the family Enterobacteriacae - 10.2%. There were methicillin-resistant strains of S. aureus in 11.8%, strains of coagulase-negative staphylococcus (MRCoNS) - 6.8%. Recurrence of the disease was observed in 7 patients. The microflora of the discharge from the fistula was represented by monocultures of S. aureus and associations of bacteria in which one of the components was methicillin-resistant strains of S. aureus. We observed differences in the contents of the microflora before reconstructive treatment of patients and in recurrence of infection. In case of recurrence of infection, the qualitative contents of the microflora changed: in 2 patients in the association of microorganisms and in 3 - in monocultures, S. aureus strains acquired resistance to ß-lactam antibiotics, new associations appeared, which were not present in primary cultures before reconstructive treatment. The study showed that the spectrum of micro-organisms in chronic osteomyelitis of the tibia, developed after osteosynthesis with a plate, varied and is subject to change and that dictates the need for microbiological monitoring to detect the etiological structure of pathogens, monitoring of antibiotic resistance of the isolated strains and rational approach to treatment of patients.


Assuntos
Fístula/microbiologia , Osteomielite/microbiologia , Ferida Cirúrgica/microbiologia , Antibacterianos , Enterobacteriaceae/isolamento & purificação , Humanos , Osteomielite/cirurgia , Staphylococcus aureus/isolamento & purificação , Tíbia/patologia
15.
BMC Infect Dis ; 19(1): 979, 2019 Nov 21.
Artigo em Inglês | MEDLINE | ID: mdl-31752702

RESUMO

BACKGROUND: Fluoroquinolones are commonly recommended as treatment for urinary tract infections (UTIs). The development of resistance to these agents, particularly in gram-negative microorganisms complicates treatment of infections caused by these organisms. This study aimed to investigate antimicrobial resistance of different Enterobacteriaceae species isolated from hospital- acquired and community-acquired UTIs against fluoroquinolones and correlate its levels with the existing genetic mechanisms of resistance. METHODS: A total of 440 Enterobacteriaceae isolates recovered from UTIs were tested for antimicrobial susceptibility. Plasmid-mediated quinolone resistance (PMQR) genes and mutations in the quinolone resistance-determining regions (QRDRs) of gyrA and parC genes were examined in quinolone-resistant strains. RESULTS: About (32.5%) of isolates were resistant to quinolones and (20.5%) were resistant to fluoroquinolones. All isolates with high and intermediate resistance phenotypes harbored one or more PMQR genes. QnrB was the most frequent gene (62.9%) of resistant isolates. Co-carriage of 2 PMQR genes was detected in isolates (46.9%) with high resistance to ciprofloxacin (CIP) (MICs > 128 µg/mL), while co-carriage of 3 PMQR genes was detected in (6.3%) of resistant isolates (MICs > 512 µg/mL). Carriage of one gene only was detected in intermediate resistance isolates (MICs of CIP = 1.5-2 µg/mL). Neither qnrA nor qnrC genes were detected. The mutation at code 83 of gyrA was the most frequent followed by Ser80-Ile in parC gene, while Asp-87 Asn mutation of gyrA gene was the least, where it was detected only in high resistant E. coli isolates (MIC ≥128 µg/mL). A double mutation in gyrA (Lys154Arg and Ser171Ala) was observed in high FQs resistant isolates (MIC of CIP < 128 µg/mL). CONCLUSION: FQs resistance is caused by interact between PMQR genes and mutations in both gyrA and parC genes while a mutation in one gene only can explain quinolone resistance. Accumulation of PMQR genes and QRDR mutations confers high resistance to FQs.


Assuntos
Antibacterianos/farmacologia , Proteínas de Bactérias/genética , Infecção Hospitalar/microbiologia , Farmacorresistência Bacteriana , Infecções por Enterobacteriaceae/microbiologia , Enterobacteriaceae/genética , Quinolonas/farmacologia , Infecções Urinárias/microbiologia , Adulto , Proteínas de Bactérias/metabolismo , Ciprofloxacino/farmacologia , Enterobacteriaceae/classificação , Enterobacteriaceae/efeitos dos fármacos , Enterobacteriaceae/isolamento & purificação , Feminino , Fluoroquinolonas/farmacologia , Humanos , Masculino , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Mutação , Plasmídeos/genética , Plasmídeos/metabolismo , Adulto Jovem
16.
J Med Microbiol ; 68(12): 1707-1715, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31661049

RESUMO

Purpose. Carbapenemase-producing Enterobacteriaceae (CPE) have become a global concern and a serious threat to human health due to their resistance to multiple antibiotics. In this study, we identified and characterized CPE for the first time in Malawi, southeastern Africa.Methodology. We investigated the possible presence of carbapenemases among a collection of 200 ceftriaxone-nonsusceptible Gram-negative clinical isolates obtained from five Malawian hospitals between January 2016 and December 2017, using both phenotypic and genotypic tests. Molecular typing of CPE was done by PFGE, multilocus sequence typing (ST) or phylogenetic grouping. Resistant plasmids were characterized by S1 PFGE, Southern blotting and conjugation assays.Results. Out of 200 isolates, we detected 16 (8 %) CPE of which all originated from one referral hospital, Kamuzu Central Hospital, in the Central part of Malawi. Of 16 isolates, seven Klebsiella pneumoniae ST340/CC258 carried bla KPC-2, two Escherichia coli ST636 (phylogroup B2) carried bla NDM-5, six E. coli ST617 (phylogroup A) and one Klebsiella variicola carried bla OXA-48. All carbapenemases were plasmid-encoded, but only bla NDM-5-carrying plasmids could be conjugated. Most isolates co-harboured other ß-lactamases and consequently exhibited a wider spectrum of resistance to commonly used antibiotics. We observed indistinguishable genetic profiles between strain types, despite originating from different wards, suggesting acquisition during admission and intra-hospital spread.Conclusion. This report strongly suggests a probable existence of highly resistant various types of CPE organisms in Malawi including KPC-2-producing K. pneumoniae ST340/CC258, a known high-risk epidemic lineage.


Assuntos
Proteínas de Bactérias/biossíntese , Enterobacteriaceae/isolamento & purificação , beta-Lactamases/biossíntese , Farmacorresistência Bacteriana Múltipla , Enterobacteriaceae/efeitos dos fármacos , Enterobacteriaceae/enzimologia , Enterobacteriaceae/genética , Genoma Bacteriano , Humanos , Testes de Sensibilidade Microbiana
17.
BMC Infect Dis ; 19(1): 841, 2019 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-31615450

RESUMO

BACKGROUND: The purposes of this study were to determine the incidence of central and peripheral venous catheter-related bacteraemias, the relationship between the suspected and final confirmed bacteraemia origins, and the differences in microbiological, epidemiological, clinical, and analytical characteristics between the groups, including evolution to death. METHODS: This was a 7-year descriptive retrospective populational study of all bloodstream infections, comparing central (CB) and peripheral (PB) venous catheter-related bacteraemias in patients older than 15 years. RESULTS: In all, 285 catheter-related bacteraemia patients, 220 with CBs (77.19%) and 65 with PBs (22.81%), were analysed among 1866 cases with bloodstream infections. The cumulative incidence per 1000 patients-day of hospital stay was 0.36 for CB and 0.106 for PB. In terms of the suspected origin, there was less accuracy in diagnosing catheter-related bloodstream infections (68. 2%) than those of other origins (78. 4%), p <  0.001. The accuracy was greater for PB (75%) than for CB (66. 2%), Coagulase-negative staphylococci were the most frequent microorganisms in both groups but occurred 1.57 times more frequently in CB (64.1%/40.6%) (p = 0.004), while Staphylococcus aureus (23. 4%/9.5%) (p = 0.02) and Enterobacteriae species (15.6%/6. 3%) (p = 0.003) were 2.5 times more frequent in PB. The CB patients stayed at the hospital for an average of 7.44 days longer than did the PB patients; more CB patients had active neoplasia (70. 4%/32.8%), more had surgery in the previous week (29. 2%/8. 3%), and fewer received adequate empirical treatment (53.9%/ 62.5%). Catheter was not removed in 8. 2% of CB and 3.7% of PB. On the other hand, the CB and PB patients had similar Pitt scores at blood extraction (median 0.89 versus 0.84 points, respectively; p = 0.8) and similar survival rates at hospital discharge (91.1% versus 90. 2%; p = 0.81). CONCLUSIONS: Central catheters were more frequent sources of bacteraemias than were peripheral catheters. There were important differences in the microbiological aetiology as well. PB patients received correct empirical antibiotic treatment more frequently and had a higher initial rate of correct determination of the suspected source of bacteraemia. Differences in the microbiological aetiology and empirical antibiotic treatment received, and probably catheter removal and time to catheter removal could explain why CB and PB patients had similar survival rates .


Assuntos
Bacteriemia/diagnóstico , Infecções Relacionadas a Cateter/diagnóstico , Cateterismo Venoso Central/métodos , Cateterismo Periférico/métodos , Bacteriemia/microbiologia , Bacteriemia/mortalidade , Infecções Relacionadas a Cateter/microbiologia , Cateteres de Demora/microbiologia , Enterobacteriaceae/isolamento & purificação , Humanos , Tempo de Internação , Estudos Retrospectivos , Staphylococcus aureus/isolamento & purificação , Taxa de Sobrevida
18.
Lett Appl Microbiol ; 69(6): 399-402, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31618795

RESUMO

The sugar glider (Petaurus breviceps) is a small, arboreal, nocturnal, gliding mammalian possum belonging to the marsupial infraclass. Exotic marsupials, including sugar gliders, are becoming popular companion pets and, consequently, the risk of potential infections that can be transmitted to humans should be investigated. Data on the role of the sugar glider as a possible carrier of pathogenic and zoonotic bacteria are scarce and fragmentary. Therefore, this study is aimed at evaluating the prevalence of potentially zoonotic bacteria (Salmonella spp., Escherichia coli, Campylobacter spp., Pseudomonas spp., Klebsiella spp., Listeria monocytogenes and Yersinia enterocolitica) in 64 sugar gliders kept as pets in Italy. The highest prevalence of infection pertained to members of the family Enterobacteriaceae, in particular Citrobacter spp. (50%), Enterobacter spp. (28·1%) and Klebsiella pneumoniae (15·6%); Pseudomonas aeruginosa was isolated from 10 out of 64 samples (15·6%). All strains of Klebsiella pneumoniae exhibited some level of resistance to multiple antimicrobials (ampicillin, amoxicillin-clavulanic acid and doxycycline). SIGNIFICANCE AND IMPACT OF THE STUDY: The results of this study show that sugar gliders may act as carriers of potentially pathogenic agents for humans and other animal species, therefore caution should be exercised in the handling and contact with these animals.


Assuntos
Farmacorresistência Bacteriana Múltipla/fisiologia , Enterobacteriaceae/efeitos dos fármacos , Enterobacteriaceae/isolamento & purificação , Marsupiais/microbiologia , Animais de Estimação/microbiologia , Animais , Enterobacteriaceae/classificação , Itália
19.
BMC Vet Res ; 15(1): 370, 2019 Oct 25.
Artigo em Inglês | MEDLINE | ID: mdl-31653209

RESUMO

BACKGROUND: Caretta caretta is the most abundant sea turtle species in the Mediterranean, and studies on this species have vastly expanded during recent years, including those investigating gut bacterial and parasitic communities. Members of these communities have been reported with variable prevalence and pathogenicity, mainly depending on their host and environment (e.g. lifespan, distribution, habitat, diet, health status and stressors). Indeed, many species commonly inhabiting the sea turtle gastrointestinal tract exhibit an opportunistic behaviour. This study aimed to provide baseline data on enterobacterial and parasitic composition, through bacteriological culture-based methods and the FLOTAC parasitological technique, in cloacal and faecal samples of 30 live Caretta caretta, examined upon their arrival at the Marine Turtle Research Centre (Portici, Italy). RESULTS: Enterobacteriaceae were isolated in 18/23 cloacal samples (78.3%), with Citrobacter and Morganella as the most common genera, followed by Proteus, Enterobacter, Providencia, and Hafnia. Parasitic elements were detected in 11/30 faecal samples (36.7%), with Enodiotrema, Rhytidodes, and Eimeria as most common genera, followed by Pachypsolus and Cymatocarpus. Additionally, Angiodyctium is reported for the first time in this host. The majority (47.8%) of sea turtles hosted exclusively Enterobacteriaceae, whereas 30.4% hosted both parasites and Enterobacteriaceae; the remaining 21.8% hosted neither of the agents. CONCLUSIONS: Bacteria and parasites evaluated in the present study are common in Mediterranean loggerhead sea turtles, with slight differences between the western and eastern basin. Although naturally present in the gastrointestinal system of free-living sea turtles, their relationship with these hosts might range from mutualism to parasitism. Indeed, members of the gut community might express their pathogenic potential in immune-compromised animals, such as those in rehabilitation facilities. Therefore, it is advisable to include in the standard work-up of rescued sea turtles a screening procedure for such opportunistic agents, in order to better evaluate the animal's health status and achieve timely intervention with appropriate treatment, thus improving rehabilitation. Furthermore, data collected from free-living sea turtles represent a starting point for investigating wild populations. However, further studies are needed to clarify the differences between sea turtle's normal gut microbiome and pathobiome.


Assuntos
Eimeria/isolamento & purificação , Enterobacteriaceae/isolamento & purificação , Trematódeos/isolamento & purificação , Tartarugas/microbiologia , Tartarugas/parasitologia , Animais , Cloaca/microbiologia , Fezes/parasitologia , Microbioma Gastrointestinal , Itália
20.
Mar Pollut Bull ; 149: 110548, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31550576

RESUMO

The present study is aimed to monitoring the ecological indicator and pathogenic microorganism diversity in pilgrimage places beach sand on the Bay of Bengal coast. The samples were collected from three locations and four different sites, and were analyzed by following standard methods. The results clearly indicates, ritual activities were highly contaminated in the beach sand qualities, and exceeded with the standard permissible limit of WHO, USEPA, EU, CPCB beach sand recreational and other contacts activities including pH (11%), TBC (100%), TCB (97%), FCB (88%), TEB (75%), E. coli (75%), disease-causing possible level of Klebsiella (84%), Shigella (75%), Salmonella (63%) and Vibrio (56%). The statistical tools were applied to find the strong evidence. The current study pointed out the major effects on the diffusion of potentially pathogenic microorganisms along the shoreline provided useful information for the setup of measures for public health protection in the Bay of Bengal coast.


Assuntos
Praias , Baías/microbiologia , Comportamento Ritualístico , /métodos , Enterobacteriaceae/isolamento & purificação , Escherichia coli/isolamento & purificação , Humanos , Índia , Klebsiella/isolamento & purificação , Salmonella/isolamento & purificação , Shigella/isolamento & purificação , Vibrio/isolamento & purificação , Microbiologia da Água
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