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1.
Int J Food Microbiol ; 336: 108902, 2021 Jan 02.
Artigo em Inglês | MEDLINE | ID: mdl-33091757

RESUMO

Routine evaluation of the slaughter process is performed by the enumeration of the aerobic colony count, Enterobacteriaceae and Salmonella spp. on the carcass through destructive or non-destructive methods. With non-destructive methods, bacteria are counted from a minimum area of 100 cm2 in different sampling sites on the pork carcasses, and the results of these investigated areas are pooled to one value for the complete carcass evaluation (a total of 400 cm2). However, the composition of the bacterial community present on the different sampling areas remains unknown. The aim of the study was to characterize the microbial population present on four areas (ham, back, jowl and belly) of eight pork carcasses belonging to two different slaughterhouses through culture-dependent (Matrix-assisted laser desorption/ionization time-of-flight Mass Spectrometry MALDI-TOF MS, combined with 16S rRNA gene sequencing) and complementary culture-independent (16S rRNA amplicon sequencing) methods. The presence of Salmonella spp. and Y. enterocolitica was additionally assessed. Using MALDI-TOF MS, Staphylococcus, Pseudomonas, and Escherichia coli were found to dominate the bacterial cultures isolated from the 8 carcasses. Based on the 16S rRNA amplicon sequencing analyses however, no specific genus clearly dominated the bacterial community composition. By using this culture-independent method, the most abundant genera in microbial populations of the ham, back, jowl and belly were found to be similar, but important differences between the two slaughterhouses were observed. Thus, present data suggests that the indigenous bacterial population of individual animals is overruled by the microbial population of the slaughterhouse in which the carcass is handled. Also, our data suggests that sampling of only one carcass area by official authorities may be appropriate for the evaluation of the hygienic status of the carcasses and therefore of the slaughter process.


Assuntos
Matadouros , Bactérias/genética , Microbiologia de Alimentos , Carne/microbiologia , Animais , Bactérias/classificação , Bactérias/isolamento & purificação , Enterobacteriaceae/isolamento & purificação , Escherichia coli/genética , Pseudomonas/genética , RNA Ribossômico 16S/genética , Salmonella/genética , Staphylococcus/genética , Suínos
2.
PLoS One ; 15(10): e0230037, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33104733

RESUMO

BACKGROUND: Bacteremia is a leading cause of mortality in developing countries, however, etiologic evaluation is infrequent and empiric antibiotic use not evidence-based. Here, we evaluated the patterns of ESBL resistance in children enrolled into a surveillance study for community acquired bacteremic syndromes across health facilities in Central and Northwestern Nigeria. METHOD: Blood culture was performed for children aged less than 5 years suspected of having sepsis from Sept 2008-Dec 2016. Blood was incubated using the BACTEC00AE system and Enterobacteriacea identified to the species level using Analytical Profile Index (API20E®). Antibiotic susceptibility profile was determined by the disc diffusion method. Real time PCR was used to characterize genes responsible for ESBL production. RESULT: Of 21,000 children screened from Sept 2008-Dec 2016, 2,625(12.5%) were culture-positive. A total of 413 Enterobacteriaceae available for analysis were screened for ESBL. ESBL production was detected in 160 Enterobacteriaceae, high resistance rates were observed among ESBL-positive isolates for Ceftriaxone (92.3%), Aztreonam (96.8%), Cefpodoxime (96.3%), Cefotaxime (98.8%) and Trimethoprim/sulfamethoxazole (90%), while 87.5%, 90.7%, and 91.9% of the isolates were susceptible to Imipenem, Amikacin and Meropenem respectively. Frequently detected resistance genes were blaTEM-83.8% (134/160), and, blaCTX-M 83.1% (133/160) followed by blaSHVgenes 66.3% (106/160). Co-existence of blaCTX-M, blaTEM and blaSHV was seen in 94/160 (58.8%), blaCTX-M and blaTEM in 118/160 (73.8%), blaTEM and blaSHV in 97/160 (60.6%) and blaCTX-M and blaSHV in 100/160 (62.5%) of isolates tested. CONCLUSION: Our results indicate a high prevalence of bacteremia from ESBL Enterobacteriaceae in this population of children. These are resistant to commonly used antibiotics and careful choice of antibiotic treatment options is critical. Further studies to evaluate transmission dynamics of resistance genes could help in the reduction of ESBL resistance in these settings.


Assuntos
Antibacterianos/farmacologia , Bacteriemia/epidemiologia , Infecções por Enterobacteriaceae/epidemiologia , Enterobacteriaceae/classificação , Resistência beta-Lactâmica , Bacteriemia/microbiologia , Pré-Escolar , Testes de Sensibilidade a Antimicrobianos por Disco-Difusão , Enterobacteriaceae/efeitos dos fármacos , Enterobacteriaceae/isolamento & purificação , Infecções por Enterobacteriaceae/microbiologia , Medicina Baseada em Evidências , Feminino , Humanos , Lactente , Espécies Introduzidas , Masculino , Nigéria/epidemiologia , Vigilância da População , Prevalência
3.
PLoS One ; 15(10): e0238939, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33017441

RESUMO

Long-term use of colistin for preventing Gram-negative bacterial infections in food animals was prohibited in Thailand in 2017, but it is permitted for short-term treatment. This study aimed to investigate association between the use of colistin for short-term treatment of infection and the emergence of colistin-resistant Enterobacteriaceae in swine. The current study was conducted at 2 selected swine farms in Thailand. Neither farm has used colistin to prevent infection for longer than 1 year. Rectal swabs were collected from the same 66 pigs at birth, and on days 7, 14, 21, 28, and 60. Colistin was used to treat sick pigs for up to 3 days. Additional rectal swabs were collected during colistin treatment. Rectal swabs were analyzed for colistin-resistant Enterobacteriaceae and the mcr-1 gene. Results revealed that colistin-resistant Enterobacteriaceae were absent at birth. Some pigs at both farms had diarrhea and received colistin treatment during days 2-27. Colistin-resistant Enterobacteriaceae were detected in 13.3-50.0% of sick and healthy pigs. No sick pigs were observed during days 28-60, and colistin was not used during that period. Colistin-resistant Enterobacteriaceae were detected in 2.8-10.0% of healthy pigs on day 28, and in 0-3.4% of healthy pigs on day 60. The mcr-1 gene was detected in 57.6% of colistin-resistant Enterobacteriaceae isolates. Short-term treatment with colistin was found to be associated with the emergence of colistin-resistant Enterobacteriaceae in swine. Colistin-resistant Enterobacteriaceae rapidly emerged after colistin use, and rapidly decreased or disappeared after its discontinuation.


Assuntos
Antibacterianos/uso terapêutico , Colistina/uso terapêutico , Infecções por Bactérias Gram-Negativas/veterinária , Sus scrofa/microbiologia , Doenças dos Suínos/tratamento farmacológico , Doenças dos Suínos/microbiologia , Animais , Animais Recém-Nascidos , Farmacorresistência Bacteriana/genética , Enterobacteriaceae/efeitos dos fármacos , Enterobacteriaceae/genética , Enterobacteriaceae/isolamento & purificação , Fazendas , Feminino , Genes Bacterianos , Infecções por Bactérias Gram-Negativas/tratamento farmacológico , Infecções por Bactérias Gram-Negativas/microbiologia , Masculino , Gravidez , Suínos , Tailândia
4.
PLoS One ; 15(9): e0236442, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32925914

RESUMO

The goal of this study was to determine the frequency of resistance to extended-spectrum cephalosporins (ESCs) in Escherichia coli and other Enterobacterales from turkeys in Canada and characterize the associated resistance determinants. Pooled fecal samples were collected in 77 turkey farms across British Columbia, Québec, and Ontario. Isolates were obtained with and without selective enrichment cultures and compared to isolates from diagnostic submissions of suspected colibacillosis cases in Ontario. Isolates were identified using MALDI-TOF and susceptibility to ESCs was assessed by disk diffusion. The presence of blaCMY, blaCTX-M, blaTEM, and blaSHV was tested by PCR. Transformation experiments were used to characterize blaCMY plasmids. Genome sequencing with short and long reads was performed on a representative sample of blaCTX-M-positive isolates to assess isolates relatedness and characterize blaCTX-M plasmids. For the positive enrichment cultures (67% of total samples), 93% (587/610) were identified as E. coli, with only a few other Enterobacterales species identified. The frequency of ESC resistance was low in E. coli isolates from diagnostic submission (4%) and fecal samples without selective enrichment (5%). Of the ESC-resistant Enterobacterales isolates from selective enrichments, 71%, 18%, 14%, and 8% were positive for blaCMY, blaTEM, blaCTX-M, and blaSHV, respectively. IncI1 followed by IncK were the main incompatibility groups identified for blaCMY plasmids. The blaCTX-M-1 gene was found repeatedly on IncI1 plasmids of the pMLST type 3, while blaCTX-M-15, blaCTX-M-55, and blaCTX-M-65 were associated with a variety of IncF plasmids. Clonal spread of strains carrying blaCTX-M genes between turkey farms was observed, as well as the presence of an epidemic blaCTX-M-1 plasmid in unrelated E. coli strains. In conclusion, Enterobacterales resistant to ESCs were still widespread at low concentration in turkey feces two years after the cessation of ceftiofur use. Although blaCMY-2 is the main ESC resistance determinant in E. coli from Canadian turkeys, blaCTX-M genes also occur which are often carried by multidrug resistance plasmids. Both clonal spread and horizontal gene transfer are involved in parallel in the spread of blaCTX-M genes in Enterobacterales from Canadian turkeys.


Assuntos
Antibacterianos/farmacologia , Cefalosporinas/farmacologia , Infecções por Enterobacteriaceae/veterinária , Enterobacteriaceae/efeitos dos fármacos , Doenças das Aves Domésticas/tratamento farmacológico , Perus/microbiologia , Animais , Antibacterianos/uso terapêutico , Cefalosporinas/uso terapêutico , Farmacorresistência Bacteriana , Enterobacteriaceae/genética , Enterobacteriaceae/isolamento & purificação , Infecções por Enterobacteriaceae/tratamento farmacológico , Escherichia coli/efeitos dos fármacos , Escherichia coli/genética , Escherichia coli/isolamento & purificação , Doenças das Aves Domésticas/microbiologia
5.
BMC Infect Dis ; 20(1): 676, 2020 Sep 16.
Artigo em Inglês | MEDLINE | ID: mdl-32938420

RESUMO

BACKGROUND: Raoultella planticola(R.planticola) is a very rare opportunistic pathogen and sometimes even associated with fatal infection in pediatric cases. Recently,the emergence of carbapenem resistance strains are constantly being reported and a growing source of concern for pediatricians. CASE PRESENTATION: We reported 4 cases of neonatal septicemia caused by Raoultella planticola. Their gestational age was 211 to 269 days, and their birth weight was 1490 to 3000 g.The R. planticola infections were detected on the 9th to 27th day after hospitalization and occured between May and June. They clinically manifested as poor mental response, recurrent cyanosis, apnea, decreased heart rate and blood oxygen, recurrent jaundice, fever or nonelevation of body temperature. The C-reactive protein and procalcitonin were elevated at significantly in the initial phase of the infection,and they had leukocytosis or leukopenia. Prior to R.planticola infection,all of them recevied at least one broad-spectrum antibiotic for 7-27d.All the R.planticola strains detected were only sensitive to amikacin, but resistant to other groups of drugs: cephalosporins (such as cefazolin, ceftetan,etc) and penicillins (such as ampicillin-sulbactam,piperacillin,etc),and even developed resistance to carbapenem. All the infants were clinically cured and discharged with overall good prognosis. CONCLUSION: Neonatal septicemia caused by Raoultella planticola mostly occured in hot and humid summer, which lack specific clinical manifestations. Pediatricians should keep in mind that R. planticola can be a potential source of neonatal sepsis and even has the potential to acquire carbapenem-resistance. Preventing outbreaks of epidemics requires early detection, timely diagnosis and treatment, and active isolation.


Assuntos
Infecções por Enterobacteriaceae/diagnóstico , Enterobacteriaceae/isolamento & purificação , Sepse Neonatal/diagnóstico , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Peso ao Nascer , Cefalosporinas/farmacologia , Cefalosporinas/uso terapêutico , Enterobacteriaceae/efeitos dos fármacos , Infecções por Enterobacteriaceae/tratamento farmacológico , Feminino , Idade Gestacional , Humanos , Lactente , Recém-Nascido , Masculino , Testes de Sensibilidade Microbiana , Sepse Neonatal/tratamento farmacológico , Piperacilina/farmacologia , Piperacilina/uso terapêutico
6.
Arch Virol ; 165(11): 2685-2687, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-32797340

RESUMO

We present here the results of the analysis of the complete genome sequence of a lytic bacteriophage, vB_ButM_GuL6, which is the first virus isolated from Buttiauxella. Electron microscopy revealed that vB_ButM_GuL6 belongs to the family Myoviridae, order Caudovirales. The genome of vB_ButM_GuL6 is a linear, circularly permuted 178,039-bp dsDNA molecule with a GC content of 43.4%. It has been predicted to contain 282 protein-coding genes and two tRNA genes, tRNA-Met and tRNA-Gly. Using bioinformatics approaches, 99 (36%) of the vB_ButM_GuL6 genes were assigned a putative function. Genome-wide comparisons and phylogenetic analysis indicated that vB_ButM_GuL6 represents a new species of the subfamily Tevenvirinae and is most closely related to Escherichia virus RB43. These phages, together with Cronobacter phages Miller, CfP1, and IME-CF2, likely form a new genus within the subfamily Tevenvirinae.


Assuntos
Enterobacteriaceae/virologia , Genoma Viral , Myoviridae/classificação , Filogenia , Crataegus/microbiologia , DNA Viral/genética , Enterobacteriaceae/isolamento & purificação , Frutas/microbiologia , Lituânia , Microscopia Eletrônica , Myoviridae/isolamento & purificação , Fases de Leitura Aberta , Análise de Sequência de DNA , Ensaio de Placa Viral , Proteínas Virais/análise , Proteínas Virais/química , Sequenciamento Completo do Genoma
7.
Int J Syst Evol Microbiol ; 70(9): 4951-4959, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32744988

RESUMO

A bacterial strain, BIT-B35T, was isolated from the gut of plastic-eating larvae of the Coleoptera insect Zophobas atratus. Its taxonomic position was determined by using a polyphasic approach. Cells were white-pigmented, Gram-stain-negative, motile short rods with terminal flagella. The 16S rRNA gene sequence (1411 bp) of strain BIT-B35T showed highest similarity (98.1%) to Escherichia fergusonii ATCC 35469T and Citrobacter koseri LMG 5519T. The results of phylogenetic analyses, based on the 16S rRNA gene, concatenated sequences of seven housekeeping genes (atpD, gyrB, infB, rpoB, pyrG, fusA and leuS) and genome sequences, placed strain BIT-B35T in a separate lineage among the family of Enterobacteriaceae. The major fatty acids were C16 : 0, C17 : 0 cyclo and C19 : 0 cyclo ω8c. The genomic DNA G+C content of strain BIT-B35T was 57.1 mol%. The chemotaxonomic data plus results of physiological and biochemical tests also distinguished strain BIT-B35T from members of other genera within the family Enterobacteriaceae. Therefore, strain BIT-B35T is considered to represent a novel species of a novel genus within the family Enterobacteriaceae, for which the name Intestinirhabdus alba gen. nov., sp. nov. is proposed. The type strain is BIT-B35T (=CGMCC 1.17042T=KCTC 72448T).


Assuntos
Besouros/microbiologia , Enterobacteriaceae/classificação , Microbioma Gastrointestinal , Filogenia , Animais , Técnicas de Tipagem Bacteriana , Composição de Bases , China , DNA Bacteriano/genética , Enterobacteriaceae/isolamento & purificação , Ácidos Graxos/química , Genes Bacterianos , Larva/microbiologia , Plásticos , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
8.
BMC Infect Dis ; 20(1): 452, 2020 Jun 29.
Artigo em Inglês | MEDLINE | ID: mdl-32600270

RESUMO

BACKGROUND: Bacterial infections are the most frequent complications in patients with malignancy, and the epidemiology of nosocomial infections among cancer patients has changed over time. This study aimed to evaluate the characteristics, antibiotic resistance patterns, and prognosis of nosocomial infections due to multidrug-resistant (MDR) bacteria in cancer patients. METHODS: This retrospective observational study analyzed cancer patients with nosocomial infections caused by MDR from August 2013 to May 2019. The extracted clinical data were recorded in a standardized form and compared based on the survival status of the patients after infection and during hospitalization. The data were analyzed using independent samples t-test, Chi-square test, and binary logistic regression. P-values < 0.05 were considered significant. RESULTS: One thousand eight patients developed nosocomial infections during hospitalization, with MDR strains detected in 257 patients. Urinary tract infection (38.1%), respiratory tract infection (26.8%), and bloodstream infection (BSI) (12.5%) were the most common infection types. Extended-spectrum ß-lactamase producing Enterobacteriaceae (ESBL-PE) (72.8%) members were the most frequently isolated MDR strains, followed by Acinetobacter baumannii (11.7%), and Stenotrophomonas maltophilia (6.2%). The results of multivariate regression analysis revealed that smoking history, intrapleural/abdominal infusion history within 30 days, the presence of an indwelling urinary catheter, length of hospitalization, and hemoglobin were independent factors for in-hospital mortality in the study population. The isolated MDR bacteria exhibited high rates of sensitivity to amikacin, meropenem, and imipenem. CONCLUSIONS: The burden of nosocomial infections due to MDR bacteria is considerably high in oncological patients, with ESBL-PE being the most predominant causative pathogen. Our findings suggest that amikacin and carbapenems actively against more than 89.7% of MDR isolates. The precise management of MDR bacterial infections in cancer patients may improve the prognosis of these individuals.


Assuntos
Antibacterianos/uso terapêutico , Infecção Hospitalar/tratamento farmacológico , Infecção Hospitalar/epidemiologia , Farmacorresistência Bacteriana Múltipla/efeitos dos fármacos , Infecções por Enterobacteriaceae/tratamento farmacológico , Infecções por Enterobacteriaceae/epidemiologia , Enterobacteriaceae/efeitos dos fármacos , Neoplasias/microbiologia , Idoso , Antibacterianos/farmacologia , China/epidemiologia , Infecção Hospitalar/microbiologia , Infecção Hospitalar/mortalidade , Enterobacteriaceae/isolamento & purificação , Infecções por Enterobacteriaceae/microbiologia , Infecções por Enterobacteriaceae/mortalidade , Feminino , Mortalidade Hospitalar , Humanos , Masculino , Pessoa de Meia-Idade , Neoplasias/mortalidade , Prevalência , Prognóstico , Estudos Retrospectivos , Fatores de Risco , Infecções Urinárias/tratamento farmacológico , Infecções Urinárias/microbiologia
9.
Int J Syst Evol Microbiol ; 70(8): 4508-4514, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32628105

RESUMO

The genus Dickeya is an important group of plant pathogens that currently comprises 10 recognized species. Although most Dickeya isolates originated from infected cultivated plants, they are also isolated from water. The genomic sequence of the Australian strain NCPPB 569T clearly established its separation from the previously characterized Dickeya species. The average nucleotide identity and digital DNA-DNA hybridization values obtained by comparing strain NCPPB 569T with strains of characterized Dickeya species were lower than 87 and 32 %, respectively, supporting the delineation of a new species. The name Dickeya poaceiphila sp. nov. is proposed for this taxon with the type strain NCPPB 569T (=CFBP 8731T). Two other strains isolated in Australia, CFBP 1537 and CFBP 2040, also belong to this species. Phenotypic and genomic comparisons enabled the identification of traits distinguishing D. poaceiphila isolates from strains of other Dickeya species.


Assuntos
Enterobacteriaceae/classificação , Filogenia , Saccharum/microbiologia , Austrália , Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano/genética , Enterobacteriaceae/isolamento & purificação , Hibridização de Ácido Nucleico , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
10.
Proc Natl Acad Sci U S A ; 117(32): 19347-19358, 2020 08 11.
Artigo em Inglês | MEDLINE | ID: mdl-32723830

RESUMO

Bacterial intracellular symbiosis (endosymbiosis) is widespread in nature and impacts many biological processes. In holometabolous symbiotic insects, metamorphosis entails a complete and abrupt internal reorganization that creates a constraint for endosymbiont transmission from larvae to adults. To assess how endosymbiosis copes-and potentially evolves-throughout this major host-tissue reorganization, we used the association between the cereal weevil Sitophilus oryzae and the bacterium Sodalis pierantonius as a model system. S. pierantonius are contained inside specialized host cells, the bacteriocytes, that group into an organ, the bacteriome. Cereal weevils require metabolic inputs from their endosymbiont, particularly during adult cuticle synthesis, when endosymbiont load increases dramatically. By combining dual RNA-sequencing analyses and cell imaging, we show that the larval bacteriome dissociates at the onset of metamorphosis and releases bacteriocytes that undergo endosymbiosis-dependent transcriptomic changes affecting cell motility, cell adhesion, and cytoskeleton organization. Remarkably, bacteriocytes turn into spindle cells and migrate along the midgut epithelium, thereby conveying endosymbionts to midgut sites where future mesenteric caeca will develop. Concomitantly, endosymbiont genes encoding a type III secretion system and a flagellum apparatus are transiently up-regulated while endosymbionts infect putative stem cells and enter their nuclei. Infected cells then turn into new differentiated bacteriocytes and form multiple new bacteriomes in adults. These findings show that endosymbiosis reorganization in a holometabolous insect relies on a synchronized host-symbiont molecular and cellular "choreography" and illustrates an adaptive feature that promotes bacteriome multiplication to match increased metabolic requirements in emerging adults.


Assuntos
Enterobacteriaceae/fisiologia , Simbiose , Gorgulhos/crescimento & desenvolvimento , Gorgulhos/microbiologia , Animais , Fenômenos Fisiológicos Bacterianos , Evolução Biológica , Sistema Digestório/microbiologia , Endófitos/genética , Endófitos/isolamento & purificação , Endófitos/fisiologia , Enterobacteriaceae/genética , Enterobacteriaceae/isolamento & purificação , Feminino , Larva/crescimento & desenvolvimento , Larva/microbiologia , Larva/fisiologia , Masculino , Metamorfose Biológica , Gorgulhos/fisiologia
11.
BMJ Case Rep ; 13(7)2020 Jul 20.
Artigo em Inglês | MEDLINE | ID: mdl-32690571

RESUMO

A 45-year-old man presented with acute sinusitis. He was treated with a 10-day course of trimethoprim/sulfamethoxazole, and a subsequent 14-day course of amoxicillin-clavulanate with no improvement in symptoms. Culture of purulent nasal secretions revealed the rare enterobacter Cedecea lapagei The patient had complete resolution of his symptoms after a 14-day course of gentamicin/dexamethasone nasal rinses. Emerging pathogens have been a timeless concern for physicians, as witnessed by the current SARS-CoV-2 outbreak. C. lapagei has been reported to cause human infection only a dozen times since its discovery, all in severely compromised patients. This is the first documented case of sinusitis reported with C. lapagei and may portend a rising prevalence of disease burden in the general population. This case demonstrates the necessity of obtaining cultures when standard antibiotics result in treatment failure.


Assuntos
Dexametasona/administração & dosagem , Infecções por Enterobacteriaceae/tratamento farmacológico , Gentamicinas/administração & dosagem , Sinusite/tratamento farmacológico , Doença Aguda , Combinação Amoxicilina e Clavulanato de Potássio/uso terapêutico , Antibacterianos/uso terapêutico , Betacoronavirus , Infecções por Coronavirus/diagnóstico , Infecções por Coronavirus/epidemiologia , Dexametasona/uso terapêutico , Quimioterapia Combinada/métodos , Enterobacteriaceae/isolamento & purificação , Infecções por Enterobacteriaceae/complicações , Gentamicinas/uso terapêutico , Humanos , Masculino , Pessoa de Meia-Idade , Pandemias , Pneumonia Viral/diagnóstico , Pneumonia Viral/epidemiologia , Sinusite/microbiologia , Resultado do Tratamento , Combinação Trimetoprima e Sulfametoxazol/uso terapêutico
12.
Appl Environ Microbiol ; 86(16)2020 08 03.
Artigo em Inglês | MEDLINE | ID: mdl-32503911

RESUMO

Traditional fermentations have been widely studied from the microbiological point of view, but little is known from the functional perspective. In this work, nitrogen fixation by free-living nitrogen-fixing bacteria was conclusively demonstrated in pozol, a traditional Mayan beverage prepared with nixtamalized and fermented maize dough. Three aspects of nitrogen fixation were investigated to ensure that fixation actually happens in the dough: (i) the detection of acetylene reduction activity directly in the substrate, (ii) the presence of potential diazotrophs, and (iii) an in situ increase in acetylene reduction by inoculation with one of the microorganisms isolated from the dough. Three genera were identified by sequencing the 16S rRNA and nifH genes as Kosakonia, Klebsiella, and Enterobacter, and their ability to fix nitrogen was confirmed.IMPORTANCE Nitrogen-fixing bacteria are found in different niches, as symbionts in plants, in the intestinal microbiome of several insects, and as free-living microorganisms. Their use in agriculture for plant growth promotion via biological nitrogen fixation has been extensively reported. This work demonstrates the ecological and functional importance that these bacteria can have in food fermentations, reevaluating the presence of these genera as an element that enriches the nutritional value of the dough.


Assuntos
Acetileno/metabolismo , Bactérias/metabolismo , Enterobacteriaceae/metabolismo , Alimentos e Bebidas Fermentados/microbiologia , Fixação de Nitrogênio , Enterobacter/isolamento & purificação , Enterobacter/metabolismo , Enterobacteriaceae/isolamento & purificação , Klebsiella/isolamento & purificação , Klebsiella/metabolismo , México , Oxirredução , Oxirredutases/análise , RNA Bacteriano/análise , RNA Ribossômico 16S/análise
13.
PLoS One ; 15(6): e0232326, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32497054

RESUMO

BACKGROUND: Bacteria that have acquired antimicrobial resistance, in particular ESBL-producing Enterobacteriaceae, are an important healthcare concern. Therefore, transmission routes and risk factors are of interest, especially for the carriage of ESBL-producing E. coli. Since there is an enhanced risk for pig slaughterhouse employees to carry ESBL-producing Enterobacteriaceae, associated with animal contact as potential risk factor, the present study investigated the occurrence of ESBL-producing Enterobacteriaceae in poultry slaughterhouse employees. Due to the higher level of resistant Enterobacteriaceae in primary poultry production than in pig production, a higher risk of intestinal colonization of poultry slaughterhouse employees was expected. RESULTS: ESBL-producing Enterobacteriaceae were detected in 5.1% (5 of 99) of the fecal samples of slaughterhouse workers. The species of these isolates was confirmed as E. coli. PCR assays revealed the presence of the genes blaCTX-M-15 (n = 2) and blaSHV-12 (n = 3) in these isolates, partly in combination with the ß-lactamase gene blaTEM-135. Participants were divided into two groups according to their occupational exposure and results indicated an increased probability of colonization with ESBL-producing Enterobacteriaceae for the group of 'higher exposure' (OR 3.7, exact 95% CI 0.6-23.5; p = 0.4). For intestinal colonization with ESBL-producing Enterobacteriaceae, a prevalence of 10% (3/30) was observed in the group of 'higher exposure' versus 2.9% (2/69) in the group of 'lower exposure'. Employees in working steps such as 'hanging' poultry in the process of slaughter and 'evisceration' seemed to have a higher risk for intestinal colonization with ESBL-producing Enterobacteriaceae compared to the group of 'lower exposure'. CONCLUSION: This study is the first of its kind to collect data on the occupational exposure of slaughterhouse workers to ESBL-producing Enterobacteriaceae in Europe. The results suggested that colonization with ESBL-producing Enterobacteriaceae is associated with occupational exposure in poultry slaughterhouses. However, the presence of ESBL-producing E. coli isolates in only 5.1% (5/99) of the tested employees in poultry slaughterhouses suggests a lower transmission risk than in pig slaughterhouses.


Assuntos
Matadouros/estatística & dados numéricos , Enterobacteriaceae/isolamento & purificação , Enterobacteriaceae/metabolismo , Intestinos/microbiologia , Exposição Ocupacional/estatística & dados numéricos , Aves Domésticas/microbiologia , beta-Lactamases/biossíntese , Adulto , Animais , Feminino , Alemanha , Humanos , Masculino , Pessoa de Meia-Idade , Exposição Ocupacional/efeitos adversos , Projetos Piloto
14.
Int J Syst Evol Microbiol ; 70(7): 4171-4178, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32552985

RESUMO

A novel Gram-stain-negative strain, designated ZYY5T, was isolated from rice roots. Results of 16S rRNA gene analysis indicated that strain ZYY5T was a member of the genus Dickeya, with a highest similarity to Dickeya zeae DSM 18068T (98.5%). The major fatty acids were summed feature 3 (C16:1 ω7c and/or C16:1 ω6c), C16:0 and summed feature 8 (C18:1 ω7c and/or C18:1 ω6c). Multi-locus sequence analysis using five concatenated genes (16S rRNA, atpD, infB, recA and gyrB) and phylogenomic analysis based on 2940 core gene sequences showed that strain ZYY5T formed a robust cluster with strains EC1, ZJU1202, DZ2Q, NCPPB 3531 and CSL RW192, while separated from the other strains of D. zeae. The orthologous average nucleotide identity (ANI) and digital DNA-DNAhybridization (dDDH) values among these six strains ranged from 96.8-99.9% and 73.7-99.8%, which supported that they were belonged to the same species. However, strain ZYY5T shared 58.4 of dDDH and 94.5% of ANI values with type strain D. zeae DSM 18068T, which were lower than the proposed species boundary cut-off for dDDH and ANI. The genomic analysis revealed that strain ZYY5T contained virulence-associated genes, which is same as the phylogenetic-related strains of the genus Dickeya. Based on the results of the polyphasic approaches, we propose that strain ZYY5T represents a novel species in the genus Dickeya, for which the name Dickeya oryzae sp. nov. (=JCM 33020 T=ACCC 61554 T) is proposed. Strains EC1, ZJU1202, DZ2Q, NCPPB 3531 and CSL RW192 should also be classified in the same genomospecies of D. oryzae same as ZYY5T.


Assuntos
Enterobacteriaceae/classificação , Oryza/microbiologia , Filogenia , Raízes de Plantas/microbiologia , Técnicas de Tipagem Bacteriana , Composição de Bases , China , DNA Bacteriano/genética , Enterobacteriaceae/isolamento & purificação , Ácidos Graxos/química , Genes Bacterianos , Hibridização de Ácido Nucleico , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
15.
J Appl Microbiol ; 129(5): 1238-1247, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-32430970

RESUMO

AIMS: To analyse and compare the effect of selection power for antimicrobial resistance (AMR) in coliforms of two kinds of ß-lactams-aminopenicillins; ampicillin (Amp) and cephalosporins; cephalexin (Cpn) and ceftiofur (Cef)-and tetracycline (Tet) using an approach based on a swine faecal microcosmos. METHODS AND RESULTS: Sixteen faecal samples from 32 pigs (mixed two by two) were treated with Amp, Cpn, Cef and Tet for 6 h (T6h) at concentrations expected to reach the animals gut when using in vivo standard doses. Controls (no drug added) were also tested. Next, samples were 1 : 100 diluted and left under the same conditions (no antimicrobial added) for further 20 h (T20h). The proportion of resistant coliform bacteria (R coliforms) to each antimicrobial was analysed just before starting the treatment (T0), at T6h and at T20h. Coselection was also studied by replica plating. Treatment for 6 h yielded significant increase in proportion of R coliforms, regardless of the drug and lack of selection pressure showed different effects at T20h depending on the antimicrobial used. Selective pressure was associated with the type of the ß-lactam with Amp selecting for significantly higher numbers of R coliforms than cephalosporins. CONCLUSIONS: AMR development was observed following short treatment, and for Amp and Tet treatment, resistance persisted 20 h beyond the interruption of treatment. An association between kind of ß-lactam and power of selection was found. SIGNIFICANCE AND IMPACT OF THE STUDY: AMR represents a threat to human health globally and antimicrobial treatment of livestock has a direct impact on this problem. Through our approach based on a swine faecal microcosmos, we demonstrated the effect on AMR development of several drugs commonly used in livestock. Cephalosporins, representing last-line antimicrobials in human medicine, exerted lower selective pressure than Amp under the conditions used and yielded higher proportion of multidrug-R strains.


Assuntos
Antibacterianos/farmacologia , Farmacorresistência Bacteriana/efeitos dos fármacos , Enterobacteriaceae/efeitos dos fármacos , Fezes/microbiologia , Ampicilina/farmacologia , Animais , Cefalosporinas/farmacologia , Enterobacteriaceae/isolamento & purificação , Suínos , Tetraciclina/farmacologia
16.
Sci Rep ; 10(1): 8113, 2020 05 15.
Artigo em Inglês | MEDLINE | ID: mdl-32415232

RESUMO

The emergence and spread of mobilized colistin resistance (mcr) genes have triggered extensive concerns worldwide. Here, we characterized the global distribution of mcr-9, a newly-identified variant of mcr, by assembling the data set of mcr-9-positive isolates from GenBank database and the literature available. Genetic features of all the mcr-9-harboring plasmids were determined by bioinformatic analysis. We showed that mcr-9 is globally distributed in 21 countries across six continents, with a wide dissemination among various species of Enterobacteriaceae strains from human, animal, food and environment. IncHI2-ST1 plasmids were found to be the predominant replicon type carrying mcr-9. Comparative genomics highlighted that IncHI2-type plasmids may also serve as a critical reservoir of mcr-9, from which different types of circulating plasmids acquired the mcr-9. Results revealed that the rcnR-rcnA-pcoE-pcoS-IS903-mcr-9-wbuC structure was consistent in most mcr-9 cassettes, suggesting a relatively unitary model involved in the mobilization of mcr-9. It is most likely that the spread of mcr-9 was mainly attributed to the conjugation and recombination events of mcr-9-carrying plasmids. In summary, our results provide a comprehensive picture of the distribution and genetic environment of mcr-9, and demonstrate the central roles played by IncHI2 plasmids in the worldwide dissemination of mcr-9.


Assuntos
Antibacterianos/farmacologia , Proteínas de Bactérias/genética , Colistina/farmacologia , Farmacorresistência Bacteriana/genética , Infecções por Enterobacteriaceae/genética , Enterobacteriaceae/genética , Plasmídeos/genética , Animais , Enzimas de Restrição do DNA , Enterobacteriaceae/classificação , Enterobacteriaceae/efeitos dos fármacos , Enterobacteriaceae/isolamento & purificação , Infecções por Enterobacteriaceae/tratamento farmacológico , Infecções por Enterobacteriaceae/microbiologia , Proteínas de Escherichia coli , Cavalos , Humanos
17.
Appl Environ Microbiol ; 86(15)2020 07 20.
Artigo em Inglês | MEDLINE | ID: mdl-32444472

RESUMO

Over a 2-year period, drag swabs of orchard soil surface and air, soil, and almond leaf samples were collected in an almond orchard adjacent to (35 m from the first row of trees) and downwind from a poultry operation and in two almond orchards (controls) that were surrounded by other orchards. Samples were evaluated for aerobic plate count, generic Escherichia coli, other coliforms, the presence of Salmonella, bacterial community structure (analyzed through sequencing of the 16S rRNA gene), and amounts of dry solids (dust) on leaf surfaces on trees 0, 60, and 120 m into each orchard. E. coli was isolated from 41 of 206 (20%) and 1 of 207 (0.48%) air samples in the almond-poultry and control orchards, respectively. Salmonella was not isolated from any of the 529 samples evaluated. On average, the amount of dry solids on leaves collected from trees closest to the poultry operation was more than 2-fold greater than from trees 120 m into the orchard or from any of the trees in the control orchards. Members of the family Staphylococcaceae-often associated with poultry-were, on average, significantly (P < 0.001) more abundant in the phyllosphere of trees closest to the poultry operation (10% of relative abundance) than in trees 120 m into the orchard (1.7% relative abundance) or from any of the trees in control orchards (0.41% relative abundance). Poultry-associated microorganisms from a commercial operation transferred a short distance into an adjacent downwind almond orchard.IMPORTANCE The movement of microorganisms, including foodborne pathogens, from animal operations into adjacent plant crop-growing environments is not well characterized. This study provides evidence that dust and bioaerosols moved from a commercial poultry operation a short distance downwind into an almond orchard and altered the microbiome recovered from the leaves. These data provide growers with information they can use to assess food safety risks on their property.


Assuntos
Microbiologia do Ar , Poeira/análise , Microbiota , Folhas de Planta/microbiologia , Microbiologia do Solo , Vento , Criação de Animais Domésticos , Animais , California , Enterobacteriaceae/isolamento & purificação , Escherichia coli/isolamento & purificação , Aves Domésticas , Prunus dulcis/microbiologia , RNA Bacteriano/análise , RNA Ribossômico 16S/análise , Salmonella/isolamento & purificação , Árvores
18.
Int J Infect Dis ; 97: 11-18, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32473388

RESUMO

OBJECTIVES: To study the molecular epidemiology of clinical metallo-ß-lactamase (MBL)-producing Enterobacteriaceae isolates in China and to evaluate the antimicrobial susceptibility of MBL-Enterobacteriaceae isolates to aztreonam-avibactam. METHODS: Bacterial speciation was determined using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. PCR was used to screen for common carbapenemase genes. Antimicrobial susceptibility testing of common clinical antibiotics and aztreonam-avibactam was performed using the standard broth microdilution method. RESULTS: A total of 161 MBL-Enterobacteriaceae isolates were included, with Klebsiella pneumoniae (n = 73, 45.4%) and Escherichia coli (n = 53, 32.9%) being the most common species. Among the 161 isolates, blaNDM (n = 151), blaIMP (n = 13), and blaVIM (n = 2) were detected, including five strains (3.1%) co-harboring two MBLs. MBL-Enterobacteriaceae isolates frequently contained two (n = 55, 34.2%) or more (n = 89, 55.3%) additional serine ß-lactamase genes (blaKPC, blaCTX-M, blaTEM, or blaSHV). Antimicrobial susceptibility testing showed that 81.4% of isolates (n = 131) were resistant to aztreonam. The rates of resistance to cefazolin, ceftazidime, ceftriaxone, cefotaxime, ampicillin-sulbactam, amoxicillin-clavulanic acid, and piperacillin-tazobactam were all over 90%. The addition of avibactam (4 µg/ml) significantly reduced the minimum inhibitory concentrations (MICs) of the aztreonam-resistant isolates by more than 8-fold (range ≤0.125 to 4 µg/ml), with a MIC50/MIC90 of ≤0.125/1 µg/ml among the 131 isolates. Overall, 96.9% (n = 156) of the total isolates were inhibited at an aztreonam-avibactam concentration of ≤1 µg/ml. Univariate and multivariate logistic regression analysis found that in patients with MBL-Enterobacteriaceae infections, the presence of pre-existing lung disease (adjusted odds ratio 8.267, 95% confidence interval 1.925-28.297; p = 0.004) was associated with a hazard effect on worse disease outcomes. CONCLUSIONS: The combined use of aztreonam-avibactam is highly potent against MBL-Enterobacteriaceae and may serve as a new candidate for the treatment of infections caused by MBL-Enterobacteriaceae in China.


Assuntos
Antibacterianos/farmacologia , Compostos Azabicíclicos/farmacologia , Aztreonam/farmacologia , Enterobacteriaceae/efeitos dos fármacos , Adulto , Idoso , Idoso de 80 Anos ou mais , Proteínas de Bactérias/genética , China , Farmacorresistência Bacteriana , Enterobacteriaceae/enzimologia , Enterobacteriaceae/genética , Enterobacteriaceae/isolamento & purificação , Escherichia coli/isolamento & purificação , Feminino , Humanos , Klebsiella pneumoniae/isolamento & purificação , Masculino , Pessoa de Meia-Idade , beta-Lactamases/biossíntese , beta-Lactamases/genética
19.
Enferm. glob ; 19(58): 257-266, abr. 2020. tab, graf
Artigo em Espanhol | IBECS | ID: ibc-195557

RESUMO

OBJETIVO: Evaluar los factores de riesgo para infecciones relacionadas con la asistencia sanitaria causadas por Enterobacteriaceae productoras de carbapenemas. MÉTODO: Este es un estudio retrospectivo de casos y controles que consistió en una muestra de 82 pacientes infectados y 164 controles, totalizando 246 pacientes. La recopilación de datos se realizó entre enero y mayo de 2017 mediante la búsqueda en el Sistema Automatizado de Control de Infecciones Hospitalarias y en los registros electrónicos de pacientes. RESULTADOS: Pacientes previamente colonizados con microorganismos gramnegativos (OR: 10.7, 95% CI: 2-60, p = 0.007), con cáncer (OR: 20.8, 95% CI: 4-120, p < 0.001), utilizando una catéter de doble luz (OR: 30.5, 95% CI: 2-382, p = 0.008), con lesión por presión (OR: 136.2, 95% CI: 11- 1623, p < 0.001) y permanecer en la Unidad de Cuidados Intensivos (OR: 1.4, 95% CI: 1.2-1.6, p <0.001) fueron más propensos a desarrollar infecciones causadas por Enterobacteriaceae productoras de carbapenemas que el grupo control. El área bajo la curva ROC mostró un buen rendimiento general (0,99; IC 95%: 0,992-0,998) del modelo de regresión logística final. CONCLUSIÓN: La colonización previa, el cáncer, el uso de catéteres de doble luz, la lesión por presión y la estadía en la UCI fueron factores de riesgo muy importantes para la adquisición de infecciones en el entorno hospitalario


OBJECTIVE: To evaluate the risk factors for healthcare-associated infections caused by Klebsiella pneumoniae carbapenemase producing Enterobacteriaceae. METHOD: This is a retrospective case-control study that consisted of a sample of 82 infected patients and 164 controls, totaling 246 patients. Data collection was performed between January and May 2017 through search in the Automated Hospital Infection Control System and in the electronic patient records. RESULTS: Patients previously colonized with gram-negative microorganisms (OR: 10.7, 95% CI: 2-60, p = 0.007), with cancer (OR: 20.8, 95% CI: 4-120, p < 0.001), using a double lumen catheter (OR: 30.5, 95% CI: 2-382, p = 0.008), with pressure injury (OR: 136.2, 95% CI: 11- 1623, p < 0.001) and Intensive Care Unit stay (OR: 1.4, 95% CI: 1.2-1.6, p <0.001) had a greater chance of developing Healthcare-associated Infections caused by KPC-producing Enterobacteriaceae than the control group. The area under the ROC curve showed a good overall performance (0.99, 95% CI: 0.992-0.998) of the final logistic regression model. CONCLUSION: Previous colonization, cancer, double lumen catheter use, pressure injury and ICU stay were very important risk factors for the acquisition of infections in the hospital environment


OBJETIVO: Avaliar os fatores de risco para infecções relacionadas à assistência à saúde causadas por Enterobactérias produtoras de Klebsiella pneumoniae carbapenemase. MÉTODO: Estudo de caso-controle, retrospectivo que foi composto por uma amostra de 82 pacientes infectados e 164 controles, totalizando 246 pacientes. A coleta de dados foi realizada entre janeiro e maio de 2017, por meio de busca no Sistema Automatizado de Controle de Infecção Hospitalar e nos prontuários eletrônicos dos pacientes. RESULTADOS: Pacientes previamente colonizados com microrganismos gram-negativos (OR: 10,7, IC 95%: 2-60, p = 0,007), com câncer (OR: 20,8, IC 95%: 4-120, p < 0,001), utilizando cateter de duplo lúmen (OR: 30,5, IC 95%: 2-382, p = 0,008), com lesão por pressão (OR: 136,2, IC 95%: 11-1623, p < 0,001) e internação na Unidade de Terapia Intensiva (OR: 1,4, IC 95%: 1,2-1,6, p < 0,001) tiveram maior chance de desenvolver infecções relacionadas à assistência à saúde causadas por Enterobactérias produtoras de Klebsiella pneumoniae carbapenemase quando comparadas ao grupo controle. A área sob a curva ROC apresentou um bom desempenho geral do modelo final de regressão logística (0,99, IC95%: 0,992-0,998). CONCLUSÃO: Colonização prévia, câncer, uso de cateter de duplo lúmen, lesão por pressão e permanência na UTI foram fatores de risco muito importantes para a aquisição de infecções no ambiente hospitalar


Assuntos
Humanos , Masculino , Feminino , Adulto , Pessoa de Meia-Idade , Idoso , Idoso de 80 Anos ou mais , Infecção Hospitalar/epidemiologia , Infecções por Klebsiella/epidemiologia , Klebsiella pneumoniae/isolamento & purificação , Infecções por Enterobacteriaceae/epidemiologia , Enterobacteriaceae/isolamento & purificação , Fatores de Risco , Farmacorresistência Bacteriana , Infecções Relacionadas a Cateter/epidemiologia , Estudos de Casos e Controles , Estudos Retrospectivos , Lesão por Pressão/epidemiologia
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