Isolation, identification, and biocontrol of antagonistic bacterium against Botrytis cinerea after tomato harvest

ABSTRACT Tomato is one of the most important vegetables in the world. Decay after harvest is a major issue in the development of tomato industry. Currently, the most effective method for controlling decay after harvest is storage of tomato at low temperature combined with usage of chemical bactericide; however, long-term usage of chemical bactericide not only causes pathogen resistance but also is harmful for human health and environment. Biocontrol method for the management of disease after tomato harvest has great practical significance. In this study, antagonistic bacterium B-6-1 strain was isolated from the surface of tomato and identified as Enterobacter cowanii based on morphological characteristics and physiological and biochemical features combined with sequence analysis of 16SrDNA and ropB gene and construction of dendrogram. Effects of different concentrations of antagonistic bacterium E. cowanii suspension on antifungal activity after tomato harvest were analyzed by mycelium growth rate method. Results revealed that antifungal activity was also enhanced with increasing concentrations of antagonistic bacterium; inhibitory rates of 1 × 105 colony-forming units (cfu)/mL antagonistic bacterial solution on Fusarium verticillioides, Alternaria tenuissima, and Botrytis cinerea were 46.31%, 67.48%, and 75.67%, respectively. By using in vivo inoculation method, it was further confirmed that antagonistic bacterium could effectively inhibit the occurrence of B. cinerae after tomato harvest, biocontrol effect of 1 × 109 cfu/mL zymotic fluid reached up to 95.24%, and antagonistic bacterium E. cowanii has biocontrol potential against B. cinerea after harvest of fruits and vegetables.

Identificación bacteriana en superficies de resina acrílica / Bacterial identification on acrylic resin surfaces

Introducción: el material usado para fabricar aparatos protésicos parciales o totales es el polimetilmetacrilato, el cual forma una superficie sólida que se encuentra en íntimo contacto con la mucosa bucal del paciente, esta superfi cie puede presentar defectos (poros, grietas e irregularidades), que se producen al momento de su elaboracióny varían según la técnica de procesado, actuando como reservorios que contribuyen a la adherencia y proliferación de microorganismos,dentro de los cuales, el más frecuentemente aislado en pacientesportadores de prótesis es Candida albicans. Objetivo: Identifi car lasbacterias presentes en la superfi cie de una resina acrílica para base dedentadura (ProBase Hot®, Ivoclar, Vivadent). Material y métodos: Se seleccionaron 10 pacientes de ambos sexos, entre 25 y 30 años de edad que acudían a la clínica de prótesis. Las impresiones de alginato de los pacientes se utilizaron para crear modelos de yeso, que luegoconfeccionaron paladares de acrílico termocurado que los pacientes llevaban por un periodo de 24 horas. Una muestra del acrílico se tomóposteriormente para fi nes de identifi cación bacteriológica. El análisisestadístico consistió en estadística descriptiva con la distribución defrecuencia y porcentajes, realizando tablas de contingencia y respuestamúltiple (Programa IBM SPSS STATISTICS 21.0). Resultados: Labacteria identifi cada mayor número de veces fue Klebsiella pneumoniae,mientras que de las aisladas en menor frecuencia correspondió tanto a Escherichia coli como a Enterobacter cloacae, Pseudomonas aeruginosa en tres oportunidades, seguido de Enterococcus faecalis, Streptococcus alfa hemolítico y Streptococcus hyicus solo un par de veces. Conclusiones: La resina acrílica usada en este estudio diopositivo a diferentes especies bacterianas y las más frecuentementeaisladas pertenecen a la familia de las enterobacterias.

Introduction: the material most commonly used to make full orpartial prosthetic dental appliances is polymethyl methacrylate, whichprovides a solid surface that is in close contact with the patient’s oralmucosa. During the production of the dental prosthetic, defects suchas holes, cracks, and other irregularities can appear on this surface(depending on the method used), which act as reservoirs that stimulatethe proliferation of microorganisms and make it easier for these toadhere to the surface. The most frequently isolated microorganism indenture wearers is Candida albicans. Objective: To identify the bacteriapresent on the acrylic resin surface of dentures bases (ProBase Hot®,Ivoclar, Vivadent). Material and methods: 10 subjects of both sexesaged between 25 and 30 years were selected from among patientsattending a prosthodontics clinic. Alginate impressions of the patientswere used to create plaster molds, which were then used to constructheat-cured acrylic resin palatal plates that the patients wore for aperiod of 24 hours. A sample of the acrylic was subsequently takenfor bacteriological identifi cation purposes. Statistical analysis wasperformed based on a descriptive analysis of frequency distributionand percentages, and crossover and multiple response tables created(IBM SPSS STATISTICS 21.0 software). Results: The most frequentlyidentifi ed bacterium in this study was Klebsiella pneumoniae, whilethe least frequently isolated were Escherichia coli and Enterobactercloacae (once each), Pseudomonas aeruginosa (in three cases),and Enterococcus faecalis, Streptococcus alpha hemolytic andStreptococcus hyicus (two each). Conclusions: The acrylic resin usedin this study tested positive for various bacterial species, the mostfrequently isolated of these belonging to the family Enterobacteriaceae.

Molecular identification of multidrug resistant Enterobacter hormaechei in Venezuela / Identificación molecular de Enterobacter hormaechei multidrogoresistente en Venezuela

Besides the importance of Enterobacter cloacae species complex as a nosocomial pathogen, little is known about the frequency of each species/genotype. Here, we describe a strain of E. hormaechei subsp. hormaechei isolated from a bronchial secretion of a patient, in the Intensive Care Unit at the General Hospital of Cumaná, Venezuela, who died due to complications of his infection. The molecular identification was done by sequencing the 16S rRNA gene and comparing it to sequences from the GenBank. This strain showed resistance to multiple families of antibiotics (MDR), and the genes blaKPC and blaVIM were detected by PCR. This is the first time E. hormaechei has been identified in Venezuela.

A pesar de la importancia de las especies del complejo Enterobacter cloacae como patógeno nosocomial, poco se conoce sobre la frecuencia de cada especie/genotipo. Aquí se describe una cepa de E. hormaechei subsp. hormaechei aislada de una secreción bronquial de un paciente internado en la Unidad de Cuidados Intensivos del Hospital General de Cumaná, Venezuela, quien murió producto de complicaciones de su infección. La identificación molecular fue hecha por secuenciación del gen ARNr 16S y porcomparación con las secuencias del GenBank. Esta cepa mostró resistencia a múltiples familias de antibióticos (MDR) y se detectaron los genes blaKPCyblaVIMpor PCR. Este es el primer reporte de E. hormaechei en Venezuela.

Isolation and characterization of arsenic resistant bacteria from wastewater

The present study proposed the isolation of arsenic resistant bacteria from wastewater. Only three bacterial isolates (MNZ1, MNZ4 and MNZ6) were able to grow in high concentrations of arsenic. The minimum inhibitory concentrations of arsenic against MNZ1, MNZ4 and MNZ6 were 300 mg/L, 300 mg/L and 370 mg/L respectively. The isolated strains showed maximum growth at 37 ºC and at 7.0 pH in control but in arsenite stress Luria Bertani broth the bacterial growth is lower than control. All strains were arsenite oxidizing. All strains were biochemically characterized and ribotyping (16S rRNA) was done for the purpose of identification which confirmed that MNZ1 was homologous to Enterobacter sp. while MNZ4 and MNZ6 showed their maximum homology with Klebsiella pneumoniae. The protein profiling of these strains showed in arsenic stressed and non stressed conditions, so no bands of induced proteins appeared in stressed conditions. The bacterial isolates can be exploited for bioremediation of arsenic containing wastes, since they seem to have the potential to oxidize the arsenite (more toxic) into arsenate (less toxic) form.

Cloning and characterization of newly isolated lipase from Enterobacter sp. Bn12

A mesophilic Enterobacter sp. Bn12 producing an alkaline thermostable lipase was isolated from soil in Tehran, Iran. The lipase gene (ELBn12) was identified from a genomic library. Sequence analysis of the DNA fragment revealed an open reading frame of 879 bp encoding a lipase with a molecular mass of 31.3 kDa. The deduced amino acid sequence showed 96% identity with a lipase of Enterobacter sp. Ag1 and the identity of their DNA sequences was 88.9%. ELBn12 belongs to the lipase subfamily I.1 and its catalytic triad consists of Ser82, Asp237 and His259. The lipase was expressed in Escherichia coli (BL21) pLysS and partially purified by anion exchange chromatography. The maximum activity of ELBn12 was obtained at temperature of 60 °C and pH 8.0 towards tricaprylin (C8) and its specific activity was around 2900 U/mg. ELBn12 was stable within a broad pH range from 6.0 to 11.0. The enzyme showed high stability in both polar and nonpolar organic solvents at 50% (v/v). The lipase activity was enhanced in the presence of 10 mM of Ca2+, Mg2+ and K+, while heavy metals (Fe3+ and Zn2+) had strong inhibitory effect. ELBn12 showed high activity in the presence of 1% (w/v) nonionic surfactants, however ionic surfactants inhibited the lipolytic activity. ELBn12 characteristics show that it has a potential to be used in various industrial processes.

Identificação de Enterobacter spp. através de técnicas convencionais e do sistema semi-automatizado Microscan (Autoscan-4) / Identification of Enterobacter spp. by conventional techniques and automated system Microscan (Autoscan-4)

Cento e noventa e cinco (195) cepas de enterobacter spp., isoladas de diversos materiais clínicos - urina, fezes, ponta de cateter, hemocultura, aspirado traqueal, fluido vaginal, ferida - foram submetidas à identificação através de provas bioquímico-fisiológicas clássicas e também, através de painéis NegCombo 20 do sistema semi-automatizado Micro-Scan-autoScan-4 (Dade Behring Inc., West Sacramento, CA, USA). As amostras eram provenientes de pacientes atendidos no Laboratório de Análises Clínicas da Faculdade de Farmácia e Bioquímica da UNOESTE, Presidente Prudente, SP e de pacientes internados no Hospital Universitário Domingos Leonardo Cerávolo, pretencente à mesma Instituição. Do total de cepas testadas, 191 (97,9%) apresentaram identificação considerada concordante entre os dois métodos utilizados; 4 cepas (2,1%) apresentaram identificação discordante, ou seja, o gênero foi diferente em cada método. Através deste estudo concluiu-se que ambos os métodos utilizados para a identificação de Enterobacter spp. apresentam vantagens e desvantagnes relacionadas ao custo, facilidade de execução, rapidez, confiabilidade entre outras características. Além disso, nossos resultados mostram que a identificação de Enterobacter spp. pode ser realizada por métodos convencionais, sem prejuízo da confiabilidade

Resistencia bacteriana del enterobacter a los antibióticos / Resistance of the enterobacter to antibiotics

Se evaluaron 113 cepas de Enterobacter spp. desde enero de 1995 hasta marzo de 1996, de seis laboratorios bacteriológicos, tres públicos y tres privados, obteniéndose 61 cepas, (53,98 por ciento) de Enterobacter aerogenes, 38 cepas (33,63 por ciento) de Enterobacter cloacae y 14 cepas (12,39 por ciento) de Enterobacter agglomerans, como resultados de cultivos de diversas fuentes, tales como secreción de heridas (34 por ciento), hemocultivos (26 por ciento), urocultivos (17 por ciento) y otros (23 por ciento). Ver Gráfico y Tabla 1. Se estudió el porcentaje de resistencia de distintas especies de Enterobacter, con discos de antibióticos, observandose una alta resistencia de las diferentes especies con ampicilina y cefalosporinas de primera generación. Se aprecia resistencia del Enterobacter cloacae al imipenem en un 3 por ciento de las cepas obtenidas, datos que concuerdan con los obtenidos a nivel nacional. Con respecto a los aminoglucósidos, llama la atención el alto porcentaje de resistencia obtenida a las diferentes especies de Enterobacter, mientras que con el ceftazidime, la resistencia del Enterobacter cloacae y del Enterobacter aerogenes cae significativamente, a diferencia de los datos obtenidos a nivel nacional, Las otras cefalosporinas (cefotaxina), cefoperazona, ceftriaxona) muestran datos variables de resistencia acordes a los datos nacionales a excepción de la cefoperazona, que muestra resistencias mayores. Se observó mayor eficacia con el uso de quinolonas, que representan el menor porcentaje de resistencias de las diferentes especies de Enterobacter, oscilando 5 y 9 por ciento