RESUMO
El objetivo del presente estudio fue evaluar la producción de blastosporas y conidios de diferentes aislados nativos de México del hongo entomopatógeno Isaria fumosorosea y de una cepa de colección mediante diferentes técnicas de propagación. En la producción de blastosporas se utilizaron 2 medios de cultivo líquidos (sumergidos), uno a base de casaminoácidos y el otro a base de peptona de colágeno como fuentes de nitrógeno, con glucosa como fuente de carbono en ambos. Para la producción de conidios, los hongos se cultivaron en agar papa dextrosa, a partir de esos cultivos se prepararon suspensiones de 1 x 10(6) conidios/ml para inocular matraces con caldo dextrosa Sabouraud, para iniciar así la fase líquida del cultivo bifásico, denominado también precultivo. Posteriormente con el precultivo y las suspensiones de conidios se inocularon bolsas con granos de arroz, que se incubaron durante 14 días para el cultivo bifásico y para la fermentación sólida, respectivamente. El aislado HIB-23 fue el que logró la más elevada concentración de blastosporas obtenida en el cultivo sumergido: 4,90 x 10(8) blastosporas/ml en el medio casaminoácidos; y en el medio con peptona de colágeno se obtuvieron 2,15 x 10(8) blastosporas/ml. La máxima producción de conidios en fermentación sólida la logró la cepa Pfr-612 (1,58 x 10(9) conidios/g), mientras que la máxima en cultivo bifásico correspondió al aislado HIB-30 (9,00 x 10(6) conidios/g). La fermentación sólida resultó ser el método más efectivo, con un promedio de 1,09 x 10(9) conidios/g, mientras que el cultivo bifásico fue el menos efectivo, con un promedio de 2,76 x 10(6) conidios/g. Para la producción de blastosporas en los medios sumergidos no se obtuvo diferencia significativa alguna.
The aim of this study was to evaluate the production of blastospores and conidia of different native isolates and a strain of Isaria fumosorosea using different propagation techniques. Two liquid culture media of casamino acids and peptone as nitrogen sources and glucose as carbon source for both media cultures were respectively used in the production of blastospores, while for the production of conidia, the fungi were grown in potato dextrose agar; from these cultures, solutions of conidia to a concentration of 1 x 10(6) per milliliter were prepared to inoculate flasks with Sabouraud dextrose broth for the liquid phase of the biphasic culture, also known as preculture. Subsequently, rice grain bags were inoculated with the preculture and the conidia solutions, which were incubated for 14 days for solid fermentation and biphasic culture, respectively. The HIB-23 isolate recorded a concentration of 4.90 x 10(8) blastospores/ml in the casamino acid medium, while a concentration of 2.15 x 10(8) blastospores/ml was obtained in the peptone collagen medium. For the Pfr-612 strain, the conidia production in solid-state fermentation was 1.58 x 10(9) conidia/g, and for HIB-30 in the biphasic culture of 9.00 x 10(6) conidia/g. Solid-state fermentation proved to be the most effective method with an average of 1.09 x 10(9) conidia/g, whereas the biphasic culture was the least effective method with 2.76 x 10(6) conidia/g; no significant difference was reported for the submerged production media.
Assuntos
Esporos Fúngicos , Hypocreales , Meios de Cultura , Fermentação , MéxicoRESUMO
The aim of this study was to evaluate the production of blastospores and conidia of different native isolates and a strain of Isaria fumosorosea using different propagation techniques. Two liquid culture media of casamino acids and peptone as nitrogen sources and glucose as carbon source for both media cultures were respectively used in the production of blastospores, while for the production of conidia, the fungi were grown in potato dextrose agar; from these cultures, solutions of conidia to a concentration of 1×106 per milliliter were prepared to inoculate flasks with Sabouraud dextrose broth for the liquid phase of the biphasic culture, also known as preculture. Subsequently, rice grain bags were inoculated with the preculture and the conidia solutions, which were incubated for 14 days for solid fermentation and biphasic culture, respectively. The HIB-23 isolate recorded a concentration of 4.90×108 blastospores/ml in the casamino acid medium, while a concentration of 2.15×108 blastospores/ml was obtained in the peptone collagen medium. For the Pfr-612 strain, the conidia production in solid-state fermentation was 1.58×109 conidia/g, and for HIB-30 in the biphasic culture of 9.00×106 conidia/g. Solid-state fermentation proved to be the most effective method with an average of 1.09×109 conidia/g, whereas the biphasic culture was the least effective method with 2.76×106 conidia/g; no significant difference was reported for the submerged production media.
Assuntos
Hypocreales , Esporos Fúngicos , Meios de Cultura , Fermentação , MéxicoRESUMO
Contamination caused by heavy metals in wastewater has a high potential of risk because they easily penetrate in to the trofic chain accumulating as organometallic compounds. In this work, the expression of mice metallothionein in E. coli (pMt-Thio) was examined as a strategy to enhance metal biosorption efficiency of bacterial biosorbents for Pb(II) and Cd(II) ions. The results showed that pMt-Thio led to significant increase in overall biosorption capacity, especially for biosorption of Pb. Isotherms and kinetic of biosorption were evaluated in this designed system. The influence of metal concentration in solution is discussed in terms of Langmuir and Freundlich isotherm and constants. The Langmuir model was found to correlate better with the experiment data. The biomass showed maximum capacities according to Langmuir adsorption model of 28.14 mgPb/gpMt-Thio and 24.27 mgCd/gpMt-Thio. The study proved that pMt-Thio is a suitable material for the removal of the heavy metal ions studied from aqueous solutions, achieving removal efficiencies higher than 90% for Pb(II) and higher than 40% for Cd(II), and could be considered as a potential material for treating effluent polluted with Cd(II) and Pb(II) ions.
Assuntos
Cádmio/metabolismo , Escherichia coli/genética , Escherichia coli/metabolismo , Chumbo/metabolismo , Metalotioneína/genética , Metalotioneína/metabolismo , Animais , Cádmio/química , Engenharia Genética/métodos , Chumbo/química , Camundongos , Eliminação de Resíduos Líquidos , Poluentes Químicos da Água/química , Poluentes Químicos da Água/metabolismo , Purificação da Água/métodosRESUMO
BACKGROUND: To understand the molecular basis of virulence variability in Entamoeba histolytica, this study presents results about differential gene expression induced by E. histolytica trophozoites in liver of hamsters in order to produce experimental amebic liver abscess (ALA) and consequently reactivate its virulence. METHODS: Amebic cultures were studied before (BALA) and after (AALA) inoculation in hamster peritoneal cavity. Markers of pathogenicity such as the rate of erythrophagocytosis, hemolytic activity, and cytotoxic effects on MDCK cell monolayers were evaluated in order to correlate these phenotypic characteristics to differential gene expression between virulent and non-virulent strains. Genotypic variability was determined by genetic polymorphism using the random-amplified polymorphic DNA (RAPD) technique, which defines the parasite genomic plasticity. mRNA differential display was used in order to identify variable transcripts levels. RESULTS: The rate of erythrophagocytosis and hemolytic activity were notably increased in AALA in comparison with BALA E. histolytica cultures, as well as the cytotoxic effect on MDCK cells. An increment in the transcription level of several mRNA was shown. CONCLUSIONS: The RAPD technique allowed us to confirm differences in number and size of polymorphic markers bands between virulent and non-virulent stages, suggesting genomic adaptability in E. histolytica. Eight different genes (membrane-bound acid phosphatase, cysteine proteinase, two different ribosomal proteins, heat shock transcription factor, ribosomal RNA, aldehyde dehydrogenase-1 and patatin-like phospholipase) were sequenced and may be associated with a biological function related to the virulence of E. histolytica. Together these findings show genomic variability between virulent and non-virulent cultures of E. histolytica.
