RESUMEN
The cell wall chitosan was extracted from fungi belonging to different taxonomic classes, namely, Benjaminiella poitrasii (Zygomycetes, dimorphic), Hanseniaspora guilliermondii, Issatchenkia orientalis, Pichia membranifaciens, and Saccharomyces cerevisiae (Ascomycetes, yeasts), and Agaricus bisporus and Pleurotus sajor-caju (Basidiomycetes). The maximum yield of chitosan was 60.89 ± 2.30 mg/g of dry mycelial biomass of B. poitrasii. The degree of deacetylation (DDA) of chitosan extracted from different fungi, as observed with 1H NMR, was in the range of 70-93%. B. poitrasii chitosan exhibited the highest DDA (92.78%). The characteristic absorption bands were observed at 3450, 1650, 1420, 1320, and 1035 cm-1 by FTIR. Compared to chitosan from marine sources (molecular weight, MW, 585 kDa), fungal chitosans showed lower MW (6.21-46.33 kDa). Further, to improve the efficacy of B. poitrasii chitosan (Bp), nanoparticles (Np) were synthesized using the ionic gelation method and characterized by dynamic light scattering (DLS). For yeast and hyphal chitosan nanoparticles (BpYCNp and BpHCNp), the average particle size was <200 nm with polydispersity index of 0.341 ± 0.03 and 0.388 ± 0.002, respectively, and the zeta potential values were 21.64 ± 0.34 and 24.48 ± 1.58 mV, respectively. The B. poitrasii chitosans and their nanoparticles were further evaluated for antifungal activity against human pathogenic Candida albicans ATCC 10231, Candida glabrata NCYC 388, Candida tropicalis ATCC 750, Cryptococcus neoformans ATCC 34664, and Aspergillus niger ATCC 10578. BpHCNps showed lower MIC90 values (0.025-0.4 mg/mL) than the chitosan polymer against the tested human pathogens. The study suggested that nanoformulation of fungal chitosan, which has low molecular weight and high % DDA, is desirable for antifungal applications against human pathogens. Moreover, chitosans as well as their nanoparticles were found to be hemocompatible and are therefore safe for healthcare applications.
Asunto(s)
Quitosano , Mucorales , Nanopartículas , Antifúngicos/farmacología , Quitosano/farmacología , Hongos , Humanos , Mucorales/químicaRESUMEN
Blakeslea trispora is known for its potential to produce an excess of carotenoids in mixed cultures of strains of opposite sex. The biosynthesis of ß-carotene in B. trispora is activated not only by sex hormone trisporic acid but also by light, especially blue light. In fungi, the most intensively investigated blue-light reception proteins are WC-1 and WC-2, and the two proteins form a transcription factor complex which is called WCC by their PAS domains. Notably, multiple genes similar to wc-1 and wc-2 have been identified and characterized in Phycomyces, Mucor, and Rhizopus. Here we report that there are four members of wc-2-like gene family in B. trispora genome: Btwc-2a, Btwc-2b, Btwc-2c, and Btwc-2d. When the mycelia were exposed to blue light, their transcription levels are regulated differentially. Except for BtWC-2b, which only has a PAS domain, the other three proteins contain both a PAS domain and a ZnF domain. BtWC-2a interacts with either BtWC-1a or BtWC-1c to form different photoreceptor complexes in yeast two-hybrid assays, which is the unique situation not yet described in other fungi. In addition, the protein-protein docking analysis by the predicted 3D structures showed that the two complexes are structurally different. These results suggested that WC proteins of B. trispora are still involved in light regulation by forming WCC and the regulation mechanism of the photobiology appears to be more complex.
Asunto(s)
Mucorales/química , Mucorales/fisiología , Factores de Transcripción/química , Factores de Transcripción/fisiología , Secuencia de Aminoácidos , Proteínas Fúngicas/química , Proteínas Fúngicas/fisiología , Regulación Fúngica de la Expresión Génica , Genoma Fúngico , Luz , Simulación del Acoplamiento Molecular , Filogenia , Conformación Proteica , Dominios Proteicos , Mapeo de Interacción de Proteínas/métodos , ARN de HongosRESUMEN
The pathogenesis of life-threatening infections caused by emerging fungal pathogens remains largely unexplored. A new study provides unprecedented evidence for the pivotal role of a new ricin-like protein toxin, named mucoricin, in causing organ necrosis and mortality in Mucorales infections.
Asunto(s)
Mucorales/química , Mucorales/patogenicidad , Micotoxinas , Animales , Humanos , Ratones , Mucormicosis/microbiología , PonzoñasAsunto(s)
Aspergillus fumigatus/genética , Galactosa/análogos & derivados , Infecciones Fúngicas Invasoras/diagnóstico , Infecciones Fúngicas Invasoras/prevención & control , Mananos/genética , Mucorales/genética , Triazoles/administración & dosificación , Aspergillus fumigatus/química , Galactosa/genética , Humanos , Infecciones Fúngicas Invasoras/microbiología , Tamizaje Masivo/métodos , Tamizaje Masivo/normas , Mucorales/química , Reacción en Cadena en Tiempo Real de la Polimerasa , Estudios RetrospectivosRESUMEN
Fungi of the order Mucorales cause mucormycosis, a lethal infection with an incompletely understood pathogenesis. We demonstrate that Mucorales fungi produce a toxin, which plays a central role in virulence. Polyclonal antibodies against this toxin inhibit its ability to damage human cells in vitro and prevent hypovolemic shock, organ necrosis and death in mice with mucormycosis. Inhibition of the toxin in Rhizopus delemar through RNA interference compromises the ability of the fungus to damage host cells and attenuates virulence in mice. This 17 kDa toxin has structural and functional features of the plant toxin ricin, including the ability to inhibit protein synthesis through its N-glycosylase activity, the existence of a motif that mediates vascular leak and a lectin sequence. Antibodies against the toxin inhibit R. delemar- or toxin-mediated vascular permeability in vitro and cross react with ricin. A monoclonal anti-ricin B chain antibody binds to the toxin and also inhibits its ability to cause vascular permeability. Therefore, we propose the name 'mucoricin' for this toxin. Not only is mucoricin important in the pathogenesis of mucormycosis but our data suggest that a ricin-like toxin is produced by organisms beyond the plant and bacterial kingdoms. Importantly, mucoricin should be a promising therapeutic target.
Asunto(s)
Mucorales/patogenicidad , Mucormicosis/patología , Micotoxinas/metabolismo , Ricina/metabolismo , Animales , Antitoxinas/inmunología , Antitoxinas/farmacología , Antitoxinas/uso terapéutico , Apoptosis , Permeabilidad Capilar , Células Cultivadas , Reacciones Cruzadas , Humanos , Hifa/química , Hifa/patogenicidad , Lectinas/metabolismo , Ratones , Mucorales/química , Mucorales/clasificación , Mucorales/genética , Mucormicosis/microbiología , Mucormicosis/prevención & control , Micotoxinas/química , Micotoxinas/genética , Micotoxinas/inmunología , Necrosis , Interferencia de ARN , Rhizopus/química , Rhizopus/genética , Rhizopus/patogenicidad , Proteínas Inactivadoras de Ribosomas/metabolismo , Ricina/química , Ricina/inmunología , Virulencia/efectos de los fármacos , Virulencia/genéticaRESUMEN
BACKGROUND: It has been reported that the genes coding for NADP-dependent glutamate dehydrogenases (NADP-GDHs) showed a cause-effect relationship with Yeast-Hypha (YH) reversible transition in a zygomycete Benjaminiella poitrasii. As YH transition is significant in human pathogenic fungi for their survival and proliferation in the host, the NADP-GDHs can be explored as antifungal drug targets. METHODS: The yeast-form specific BpNADPGDH I and hyphal-form specific BpNADPGDH II of B. poitrasii were purified by heterologous expression in E. coli BL-21 cells and characterized. The structural analogs of L-glutamate, dimethyl esters of isophthalic acid (DMIP) and its derivatives were designed, synthesized and screened for inhibition of NADP-GDH activity as well as YH transition in B. poitrasii, and also in human pathogenic Candida albicans strains. RESULTS: The BpNADPGDH I and BpNADPGDH II were found to be homo-hexameric proteins with native molecular mass of 282 kDa and 298 kDa, respectively and subunit molecular weights of 47 kDa and 49 kDa, respectively. Besides the distinct kinetic properties, BpNADPGDH I and BpNADPGDH II were found to be regulated by cAMP-dependent- and Calmodulin (CaM) dependent- protein kinases, respectively. The DMIP compounds showed a more pronounced effect on H-form specific BpNADPGDH II and inhibited YH transition as well as growth in B. poitrasii and C. albicans strains. CONCLUSION: The present study will be useful to design and develop antifungal drugs against dimorphic human pathogens using glutamate dehydrogenase as a target. SIGNIFICANCE: Glutamate dehydrogenases can be explored as a target against human pathogenic fungi.
Asunto(s)
Antifúngicos/farmacología , Inhibidores Enzimáticos/farmacología , Glutamato Deshidrogenasa (NADP+)/antagonistas & inhibidores , Glutamato Deshidrogenasa (NADP+)/metabolismo , Mucorales/enzimología , Animales , Antifúngicos/química , Diseño de Fármacos , Inhibidores Enzimáticos/química , Proteínas Fúngicas/antagonistas & inhibidores , Proteínas Fúngicas/química , Proteínas Fúngicas/metabolismo , Glutamato Deshidrogenasa (NADP+)/aislamiento & purificación , Humanos , Mucorales/química , Mucorales/efectos de los fármacos , Mucorales/metabolismo , Mucormicosis/tratamiento farmacológico , Mucormicosis/microbiología , Fosforilación/efectos de los fármacos , OvinosRESUMEN
Fungal chitosan (FCt) from Amylomyces rouxii, with 88.7% deacetylation degree and 112.4 kDa molecular weight, was utilized for nanoparticles (NPs) formation via ionic gelation. FCt-NPs were employed as carriers for curcumin (CUR) to augment its availability and anticancer bioactivity. The synthesis of CUR/FCt-NPs composite was succeeded as evidenced from their FTIR spectra. The scanning micrographs of synthesized CUR/FCt-NPs indicated their spherical shapes and well-distribution; they had average diameters of 115 ± 21 nm and positive zeta potentials of +33.8 mV. The NPs loading capacity for CUR was 21.6% and the encapsulation efficiency reached 83.8%. The CUR was vastly released in the beginning 5 h then gradually released up to 90 h, with higher release in pH 5.2 than in pH 7.0. The treatment of cancer cells, HCT-116 and A-549, with CUR/FCt NPs lead to time-dependent decrement of cells' viability; the dead cells were 67.6% from HCT-116 and 73.8% from A-546 after 96 h of exposure. Fluorescent imaging indicated that most cancer cells entered the apoptosis phase after treatment with 150 µM of CUR/FCt-NPs for 72 h. The efficiency of FCt-NPs was proved as carriers for loading CUR and augmenting its anticancer activity toward human cancer cells, using these natural and biosafe agents.
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Antineoplásicos/farmacología , Apoptosis/efectos de los fármacos , Quitosano/química , Curcumina/farmacología , Portadores de Fármacos/química , Mucorales/química , Células A549 , Células HCT116 , Humanos , Nanopartículas/química , Neoplasias/tratamiento farmacológicoRESUMEN
Fungal chitosan (ACT) extraction from Amylomyces rouxii, its transforming into nano-form, loading with fluconazole (Flu) and evaluation of synthesized nanoconjugates against drug-resistant (DR) Candida spp., were investigated. The produced ACT was characterized with 112.4â¯kDa molecular weight and 88.7% deacetylation degree. Synthesis of chitosan nanoparticles (NACT), and loading them with Flu were succeeded, using ionic gelation protocol, to generate stable Flu/NACT nanoconjugate' particles with mean size of 82â¯nm and zeta potential of +3.36â¯mV. The NACT entrapment efficiency was 78.7% and the drug loading capacity was 60.2%. Flu slowly released from NACT during the first 5â¯h, then release dramatically increased to the maximum (94.8%) after 12â¯h. The infra-red spectrum of Flu/NACT nanoconjugates confirmed the strong cross-linkage between their molecules. The antimycotic activity of NACT and Flu/NACT was proved against DR strains of C. albicans (2 strains), C. parapsilosis and C. glabrata, using qualitative and quantitative assays; Flu/NACT exhibited significant powerful activity, which was confirmed via observations with scanning microscopy. Finished cotton textiles with Flu/NACT had augmented potentiality for inhibiting challenged DR Candida spp., using in vitro assay. Accordingly, the synthesis and application of Flu/NACT nanoconjugates was astoundingly recommended for controlling DR Candida spp.
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Candida/crecimiento & desarrollo , Quitosano , Farmacorresistencia Fúngica/efectos de los fármacos , Fluconazol , Polisacáridos Fúngicos , Mucorales/química , Quitosano/química , Quitosano/farmacología , Preparaciones de Acción Retardada/química , Preparaciones de Acción Retardada/farmacología , Fluconazol/química , Fluconazol/farmacología , Polisacáridos Fúngicos/química , Polisacáridos Fúngicos/farmacologíaRESUMEN
A novel bio-nanocomposite was prepared by the combination of fungal biomass and Fe3O4 magnetic nanoparticles. The result of XRD and EDAX analysis indicated that Fe3O4 Actinomucor sp. bio-nanoadsorbent was prepared. Our experiments showed that the adsorption kinetics and isotherm of this material comply with the pseudo-second-order and the Langmuir models, respectively. The maximum adsorption capacity (qmax) of this novel bio-nanoadsorbent was obtained as 29.49 mg/g. The thermodynamic analysis revealed that the adsorption of Cd2+ is spontaneous and exothermic. The optimum temperature, initial concentration, contact time and pH for adsorption system of cadmium were about 45 °C, 400 mg/L, 120 min and 7, respectively. Pretreatment of adsorbent by NaOH and SDS significantly increased cadmium adsorption capacity. SEM images showed that Fe3O4 nanoparticles were immobilized successfully on the fungus cell surface. Contribution of the carboxyl, hydroxyl, amine and Fe-O functional groups of the bio-nanoadsorbent in the binding to cadmium ions was revealed by FTIR analysis. Results from regeneration studies indicated reusability of the adsorbent up to 91%. According to experimental results, it could be claimed that bio-nanocomposite of Fe3O4-Actinomucor sp. is a novel efficient adsorbent for removal of metal ions from aqueous solutions, and hence it has potential to be used in the environmental pollution cleanup programs.
Asunto(s)
Cadmio/química , Nanopartículas de Magnetita/química , Mucorales/química , Contaminantes Químicos del Agua/química , Purificación del Agua , Adsorción , Agua/químicaRESUMEN
Mucorales are fungi with increasing importance in the clinics. Infections take a rapidly progressive course resulting in high mortality rates. The ergosterol biosynthesis pathway and sterol composition are of interest, since they are targeted by currently applied antifungal drugs. Nevertheless, Mucorales often exhibit resistance to these drugs, resulting in therapeutic failure. Here, sterol patterns of six clinically relevant Mucorales (Lichtheimia corymbifera, Lichtheimia ramosa, Mucor circinelloides, Rhizomucor pusillus, Rhizopus arrhizus, and Rhizopus microsporus) were analysed in a targeted metabolomics fashion after derivatization by gas chromatography-mass spectrometry. Additionally, the effect of posaconazole (POS) treatment on the sterol pattern of R. arrhizus was evaluated. Overall, fifteen different sterols were detected with species dependent variations in the total and relative sterol amount. Sterol analysis from R. arrhizus hyphae confronted with sublethal concentrations of posaconazole revealed the accumulation of 14-methylergosta-8,24-diene-3,6-diol, which is a toxic sterol that was previously only detected in yeasts. Sterol content and composition were further compared to the well-characterized pathogenic mold Aspergillus fumigatus. This work contributes to a better understanding of the ergosterol biosynthesis pathway of Mucorales, which is essential to improve antifungal efficacy, the identification of targets for novel drug design, and to investigate the combinatorial effects of drugs targeting this pathway.
Asunto(s)
Aspergillus fumigatus/química , Farmacorresistencia Fúngica , Mucorales/química , Esteroles/química , Antifúngicos/farmacología , Aspergillus fumigatus/efectos de los fármacos , Ergosterol/análogos & derivados , Ergosterol/aislamiento & purificación , Cromatografía de Gases y Espectrometría de Masas , Metabolómica , Estructura Molecular , Mucorales/clasificación , Mucorales/efectos de los fármacos , Especificidad de la Especie , Triazoles/farmacologíaRESUMEN
Rhino-orbito-cerebral mucormycosis from dental origin is an acute infection caused by opportunistic fungi belonging to the order of Mucorales, which affects mainly diabetic and immunocompromised patients.We report the case of a 63-year old diabetic man who performed a dental extraction on himself by his own means and subsequently developed a rhino-orbito-cerebral mucormycosis with cutaneous and palatal affection. The species isolated in the mycological culture was Rhizopus sp.
Asunto(s)
Diabetes Mellitus/microbiología , Huésped Inmunocomprometido , Mucorales/química , Mucormicosis/microbiología , Diabetes Mellitus/inmunología , Humanos , Masculino , Mucormicosis/diagnósticoRESUMEN
Resumen La mucormicosis es una infección aguda causada por hongos oportunistas pertenecientes al orden de los mucorales, que afecta principalmente a pacientes diabéticos e inmunosuprimidos. Se reporta el caso de un hombre diabético de 63 años de edad, que se extrajo una pieza dental por sus propios medios y, posteriormente, desarrolló una mucormicosis rino-órbito-cerebral con afección cutánea y palatina. La especie aislada mediante cultivos micológicos fue Rhizopus sp.
Abstract Rhino-orbito-cerebral mucormycosis from dental origin is an acute infection caused by opportunistic fungi belonging to the order of Mucorales, which affects mainly diabetic and immunocompromised patients. We report the case of a 63-year old diabetic man who performed a dental extraction on himself by his own means and subsequently developed a rhino-orbito-cerebral mucormycosis with cutaneous and palatal affection. The species isolated in the mycological culture was Rhizopus sp.
Asunto(s)
Humanos , Masculino , Huésped Inmunocomprometido , Diabetes Mellitus/microbiología , Mucorales/química , Mucormicosis/microbiología , Diabetes Mellitus/inmunología , Mucormicosis/diagnósticoRESUMEN
Most fungi are known to synthesize siderophores under iron limitation. However, arbuscular mycorrhizal fungi (AM fungi) have so far not been reported to produce siderophores, although their metabolism is iron-dependent. In an approach to isolate siderophores from AM fungi, we have grown plants of Tagetes patula nana in the presence of spores from AM fungi of the genus Glomus (G. etunicatum, G. mossae & unidentified Glomus sp.) symbiotically under iron limitation and sterile conditions. A siderophore was isolated from infected roots after 2-3 weeks of growth in pots containing low-iron sand with Hoagland solution. HPLC analysis of the root cell lysate revealed a peak at a retention time of 6.7 min which showed iron-binding properties in a chrome azurol S test. The compound was isolated by preparative HPLC and the structure was determined by high resolution electrospray FTICR-MS and GC/MS analysis of the hydrolysis products. From an observed absolute mass to charge ratio (m/z) of 401.11925 [M+H]+ with a relative mass error of ∆ = 0.47 ppm an elemental composition of C16H21N2O10 [M+H]+ was derived, suggesting a molecular weight of 400 Da for glomuferrin. Corresponnding ion masses of m/z 423.10 and m/z 439.06 were asigned to the Na-adduct and K-adduct respectively. A mass of 455.03836 confirmed an Fe- complex with an elemental composition of C16H19N2O10Fe (∆ = 0.15 ppm). GC/MS analysis of the HCl lysate (6 N HCL, 12 h) revealed 1,4 butanediamine. Thus the proposed structure of the isolated siderophore from Glomus species consisted of 1,4 butanediamine amidically linked to two dehydrated citrate residues, similar to the previously identified bis-amidorhizoferrin. Thus, the isolated siderophore (glomuferrin) is a member of the rhizoferrin family previously isolated from fungi of the Mucorales (Zygomycetes).
Asunto(s)
Compuestos Férricos/aislamiento & purificación , Hierro/química , Mucorales/química , Micorrizas/química , Putrescina/aislamiento & purificación , Sideróforos/aislamiento & purificación , Cromatografía Líquida de Alta Presión , Compuestos Férricos/química , Peso Molecular , Mucorales/crecimiento & desarrollo , Mucorales/metabolismo , Micorrizas/crecimiento & desarrollo , Micorrizas/metabolismo , Putrescina/química , Sideróforos/química , Esporas Fúngicas/crecimiento & desarrollo , Esporas Fúngicas/metabolismo , Simbiosis , Tagetes/microbiologíaRESUMEN
Apocarotenoids are metabolites originated by degradation of carotenes through the loss of carbon atoms placed at the side chain of their structure as consequence of oxydative reactions. We present here the first review of apocarotenoids in the fungi mucorales Phycomyces blakesleeanus, Blakeslea trispora and Mucor mucedo. This review is divided into two parts: the first one presents their structures and sources, whereas the second part is dedicated to their chemical synthesis.
Asunto(s)
Carotenoides/química , Carotenoides/síntesis química , Mucorales/química , Estructura MolecularRESUMEN
Lycopene is an abundant natural carotenoid pigment with several biological functions (well-known for its antioxidant properties) which is under intensive investigation in recent years. Lycopene chemistry, its natural distribution, bioavailability, biological significance, and toxicological effects are briefly outlined in the first part of this review. The second, major part, deals with various modern downstream processing techniques, which are assessed in order to identify promising approaches for the recovery of lycopene and of similar lipophilic compounds. Natural lycopene is synthesized in plants and by microorganisms, with main representatives of these two categories (for industrial production) tomato and its by-products and the fungus Blakeslea trispora, respectively. Currently, there is a great deal of effort to develop efficient downstream processing for large scale production of natural-origin lycopene, with trends strongly indicating the necessity for "green" and mild extraction conditions. In this review, emphasis is placed on final product safety and ecofriendly processing, which are expected to totally dominate in the field of natural-origin lycopene extraction and purification.
Asunto(s)
Anticarcinógenos/química , Anticarcinógenos/aislamiento & purificación , Antioxidantes/química , Antioxidantes/aislamiento & purificación , Carotenoides/química , Carotenoides/aislamiento & purificación , Animales , Anticarcinógenos/metabolismo , Anticarcinógenos/toxicidad , Antioxidantes/metabolismo , Antioxidantes/toxicidad , Bacterias/química , Disponibilidad Biológica , Carotenoides/metabolismo , Carotenoides/toxicidad , Citrullus/química , Enzimas/metabolismo , Frutas/química , Hongos/química , Semivida , Humanos , Absorción Intestinal , Licopeno , Solanum lycopersicum/química , Microondas , Mucorales/química , Presión , Solventes/química , Solventes/clasificación , Tensoactivos/metabolismo , Ondas Ultrasónicas , Verduras/químicaRESUMEN
Asiatic acid (1) is a natural triterpenoid isolated from Centella asiatica. This paper reports the microbial transformation of asiatic acid by an endophytic fungus Umbelopsis isabellina to obtain derivatives potentially useful for further studies. Incubation of asiatic acid with U. isabellina afforded two derivatives 2α,3ß,7ß, 23-tetrahydroxyurs-12-ene-28-oic acid (2) and 2α,3ß,7ß,23-tetrahydroxyurs-11-ene-28,13-lactone (3). The structures of these compounds were elucidated by spectral data. Compound 3 has formed an unusual lactone. These two products are new compounds. The possible transformation passway was also discussed.
Asunto(s)
Centella/química , Lactonas/aislamiento & purificación , Triterpenos Pentacíclicos , Triterpenos/aislamiento & purificación , Biotransformación , Hidroxilación , Lactonas/química , Estructura Molecular , Mucorales/química , Triterpenos Pentacíclicos/química , Triterpenos Pentacíclicos/aislamiento & purificación , Triterpenos Pentacíclicos/metabolismo , Triterpenos/químicaRESUMEN
A new aromatic ester, pilobolusate (1), four new depsidones, pilobolusones A-D (2-5), five known depsidones, (6-10), and ergosterol were isolated from the fungus Pilobolus heterosporus. Their structures were established on the basis of spectroscopic data. Compounds 2 and 4-9 showed cytotoxicity against three cancer cell lines (KB, MCF-7, and NCI-H187) with IC50 values in the range of 9.94-97.42 µM. In addition, compounds 2, 5, 9, and 10 exhibited antimalarial activity against Plasmodium falciparum with IC50 values ranging from 3.67-23.56 µM.
Asunto(s)
Antimaláricos/química , Depsidos/química , Lactonas/química , Mucorales/química , Animales , Antimaláricos/aislamiento & purificación , Antimaláricos/farmacología , Línea Celular Tumoral , Chlorocebus aethiops , Depsidos/aislamiento & purificación , Depsidos/farmacología , Lactonas/aislamiento & purificación , Lactonas/farmacología , Células MCF-7 , Resonancia Magnética Nuclear Biomolecular , Plasmodium falciparum , Células VeroRESUMEN
Glyphosate (N-phosphonomethyl glycine) is registered as a herbicide for many food and non-food crops, as well as non-crop areas where total vegetation control is desired. Glyphosate influences the soil mycobiota; however, the possible effect of glyphosate residues in animal feed (soybean, corn, etc.) on animal mycobiota is almost unknown. Accordingly, the present study was initiated to investigate the mycological characteristics of dairy cows in relationship to glyphosate concentrations in urine. A total of 258 dairy cows on 14 dairy farms in Germany were examined. Glyphosate was detected in urine using ELISA. The fungal profile was analyzed in rumen fluid samples using conventional microbiological culture techniques and differentiated by MALDI-TOF mass spectrometry. LPS-binding protein (LBP) and antibodies (IgG1, IgG2, IgA, and IgM) against fungi were determined in blood using ELISA. Different populations of Lichtheimia corymbifera, Lichtheimia ramosa, Mucor, and Rhizopus were detected. L. corymbifera and L. ramosa were significantly more abundant in animals containing high glyphosate (>40 ng/ml) concentrations in urine. There were no significant changes in IgG1 and IgG2 antibodies toward isolated fungi that were related to glyphosate concentration in urine; however, IgA antibodies against L. corymbifera and L. ramosa were significantly lower in the higher glyphosate groups. Moreover, a negative correlation between IgM antibodies against L. corymbifera, L. ramosa, and Rhizopus relative to glyphosate concentration in urine was observed. LBP also was significantly decreased in animals with higher concentrations of glyphosate in their urine. In conclusion, glyphosate appears to modulate the fungal community. The reduction of IgM antibodies and LBP indicates an influence on the innate immune system of animals.
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Antifúngicos/farmacología , Biota/efectos de los fármacos , Glicina/análogos & derivados , Mucorales/efectos de los fármacos , Mucorales/crecimiento & desarrollo , Rumen/microbiología , Animales , Anticuerpos Antifúngicos/sangre , Antifúngicos/análisis , Proteínas Portadoras/sangre , Bovinos , Ensayo de Inmunoadsorción Enzimática , Alemania , Glicina/análisis , Glicina/farmacología , Mucorales/química , Mucorales/aislamiento & purificación , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , Orina/química , GlifosatoRESUMEN
Rapid and efficient extraction of bioactive compounds from biomass for sample analysis presents one of the major challenges in high-productive strain screening and bioprocess development. In this work, a high-throughput method based on a new setup was developed for the extraction of ß-carotene from Blakeslea trispora. After the cell was completely disrupted by high-throughput machinery grinding, ß-carotene was completely extracted by chloroform and dimethylsulfoxide (1:1, v/v). According to the experimental results, the newly developed high-throughput extraction (HTPE) method could simultaneously process 96 samples within 30 Min, and only 5-mL solvent was used for each sample. The sufficient extraction of ß-carotene by the HTPE method, which was confirmed by the conventional reference extraction method, further demonstrated that the newly developed HTPE method was more efficient and economical than the methods developed previously for the extraction of ß-carotene.
Asunto(s)
Fraccionamiento Químico/métodos , Ensayos Analíticos de Alto Rendimiento , Mucorales/química , beta Caroteno/aislamiento & purificación , Factores de TiempoRESUMEN
Blakeslea trispora powder that contains 1.9 % lycopene was tested for its anti-aging effect on adult mice. 48 adult mice were administered with the powder at 0, 267, 534, 1,068 mg lycopene kg(-1) body daily for 30 days. The body weight, hematology, clinical chemical and antioxidant activities in major organs of adult mice were measured. The powder had no effect on the body weight, hematology, clinical chemical parameters of adult mice but improved the antioxidant activities in major organs of adult mice. Increased activities of superoxide dismutase, catalase and glutathione peroxidase and a decreased amount of malondialdehyde in liver, brain, kidney and skin of adult mice when a high-dose of the B. trispora powder was administered, suggests that it has the ability to enhance the antioxidation system and improve the anti-aging abilities of adult mice.