Filamentous fungal characterizations by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

Matrix-assisted laser desorption/ionization time-of-flight intact cell mass spectrometry (MALDI-TOF ICMS) is coming of age for the identification and characterization of fungi. The procedure has been used extensively with bacteria. UV-absorbing matrices function as energy mediators that transfer the absorbed photoenergy from an irradiation source to the surrounding sample molecules, resulting in minimum fragmentation. A surprisingly high number of fungal groups have been studied: (i) the terverticillate penicillia, (ii) aflatoxigenic, black and other aspergilli, (iii) Fusarium, (iv) Trichoderma, (iv) wood rotting fungi (e.g. Serpula lacrymans) and (v) dermatophytes. The technique has been suggested for optimizing quality control of fungal Chinese medicines (e.g. Cordyceps). MALDI-TOF ICMS offers advantages over PCR. The method is now used in taxonomic assessments (e.g. Trichoderma) as distinct from only strain characterization. Low and high molecular mass natural products (e.g. peptaibols) can be analysed. The procedure is rapid and requires minimal pretreatment. However, issues of reproducibility need to be addressed further in terms of strains of species tested and between run variability. More studies into the capabilities of MALDI-TOF ICMS to identify fungi are required

Interactions between Penicillium brevicompactum/Penicillium expansum and Acinetobacter calcoaceticus isolated from drinking water in biofilm development and control

Available online 13 October 2022Bacteria and filamentous fungi (ff) are commonly encountered in biofilms developed in drinking water (DW) distribution systems (DWDS). Despite their intimate ecological relationships, researchers tend to study bacteria and ff separately. This work assesses the impact of bacteria-ff association in biofilm formation and tolerance to chlorination. One strain of Acinetobacter calcoaceticus isolated from DW was used as a model bacterium. Penicillium brevicompactum and P. expansum isolated from DW were the ff selected. Single species and inter-kingdom adhesion and biofilm formation occurred under two shear stress () conditions (0.05 and 1.6 Pa). The sessile structures were further characterized in terms of biomass production, respiratory activity and structure. The results showed that 1.6 Pa of shear stress and A. calcoaceticus-ff association favoured biofilm production. Inter-kingdom biofilms produced more biomass than A. calcoaceticus single species and reduced A. calcoaceticus susceptibility to disinfection, particularly to high sodium hypochlorite (SHC) concentrations. In addition, P. brevicompactum formed single species biofilms highly resistant to removal and inactivation by SHC. The presence of P. brevicompactum or P. expansum in inter-kingdom biofilms significantly decreased SHC removal and inactivation effects in comparison to the bacterial biofilms alone, proposing that using bacteria to form biofilms representative of DWDS can provide inaccurate conclusions, particularly in terms of biofilm production and susceptibility to disinfection.This work was financially supported by LA/P/0045/2020 (ALiCE), UIDB/00511/2020 and UIDP/00511/2020 (LEPABE) - funded by national funds through the FCT/MCTES (PIDDAC); Projects Germirrad - POCI-01-0247-FEDER-072237; PRESAGE - Aquatic/0007/2020, funded by FEDER funds through COMPETE2020 – Programa Operacional Competitividade e Internacionalizaçao ˜ (POCI) and by national funds (PIDDAC) through FCT/MCTES; Project “HealthyWaters – Identification, Elimination, Social Awareness and Education of Water Chemical and Biological Micropollutants with Health and Environmental Implications”, with reference NORTE-01-0145-FEDER-000069, supported by Norte Portugal Regional Operational Programme (NORTE 2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF). This study was further supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UIDB/04469/2020 unit and by LABBELS – Associate Laboratory in Biotechnology, Bioengineering and Microelectromechnaical Systems, LA/P/0029/2020.info:eu-repo/semantics/publishedVersio

The effects of sodium hypochlorite on the control of inter-kingdom biofilm formation by drinking water-isolated microorganisms

Biofilms in drinking water distribution systems (DWDS) are responsible for several undesirable effects in water. One of the main drawbacks is their potential to protect pathogens from stress conditions. Microbial interactions in biofilms can benefit the survival of co existing microorganisms, including the increased resistance to antimicrobials. Chlorine disinfection is the main widespread strategy used in DWDS for microbial control. Even if new and alternative strategies are being developed, it is conceivable that the future strategies still persist with chlorine due to economic and safety aspects. Therefore, the understanding on the efficacy of chlorine against biofilms is of utmost importance in order to improve the current strategies. The purpose of this work was to assess the effects of sodium hypochlorite (SHC) on the control of single and dual species biofilm formation by selected filamentous fungi (Penicillium expansum and Penicillium brevicompactum) and bacterium (Acinetobacter calcoaceticus) isolated from DWDS. Biofilms were developed during 48 h in 96 wells microtiter plates under two hydrodynamic conditions (25 and 150 rpm). The effects of SHC at several concentrations (0.1, 0.5, 1, 10 and 100 mg/L) was tested. The biofilm control was evaluated using crystal violet (removal) and resazurin (inactivation) dyes. The results shown that, P.brevicompactum biofilms were extremely resistant to disinfection when compared with single species biofilms of P.expansum and dual species biofilms of P.brevicompactum A.calcoaceticus. The association of A.calcoaceticus with both fungi seems beneficial, since the dual species biofilms were more resistant to disinfection. The inactivation and removal occurred for high SHC concentrations. However, total biofilm control was not achieved

First Report of Colletotrichum gloeosporioides Causing Anthracnose of Tejocote (Crataegus gracilior) Fruits in Mexico

Crataegus, is a genus classified in family Rosaceae and includes several tree species commonly called Tejocote that are widely cultivated for their pome fruits in Mexico. During fall of 2014, 2015, and 2016, severe symptoms of anthracnose were observed on approx. 60% of tejocote (Crataegus gracilior) fruits in an orchard located in Tulancingo, Oaxaca, Mexico. Affected fruits showed sunken, prominent, dark brown to black necrotic lesions, and were exuding salmon spore masses. To isolate the fungus, small pieces from tissue adjacent to the lesions of 10 symptomatic fruits were excised and surface disinfested by immersion in a 1% sodium hypochlorite solution for 2 min, rinsed three times in sterile distilled water, placed in Petri plates containing potato dextrose agar (PDA), and incubated at 25ºC for 5 to 7 days in darkness. Mycelial plugs were excised from the edge of the actively growing fungal colony and aseptically transferred to fresh PDA medium and incubated at 25°C for 6 days. Five monoconidial cultures were obtained by transferring germinated spores to Petri plates with fresh PDA. One isolate was selected as representative for morphological and molecular identification. Colonies of pure cultures exhibited greyish-white aerial mycelium and abundant salmon-pink conidial masses. Conidia (n= 100) were subcylindrical, hyaline, straight, one-celled, with rounded ends, measuring 13.6 to 17.7 × 4.4 to 5.9 μm. Conidial appressoria were ovoid and brown to dark brown. Based on morphological characteristics, the fungus was identified within the Colletotrichum gloeosporioides species complex (Weir et al. 2012). The isolate was designated UACH-177 and deposited in the Culture Collection of Phytopathogenic Fungi at the Chapingo Autonomous University. For molecular identification, the ITS region (White et al. 1990), and fragments of (Apn2) (Rojas et al. 2010), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and β-tubulin 2 (TUB2) genes (Weir et al. 2012) were amplified by PCR, and sequenced. The sequences were deposited in GenBank (Accessions numbers ITS:MG821312; Apn2:MG821310; GAPDH:MG821311; and TUB2:MG821313). A phylogenetic analysis using Bayesian inference and including published ITS, Apn2, GAPDH, and TUB2 data for C. gloeosporioides and other Colletotrichum species was performed. The phylogenetic analysis showed the sequences were grouped into the clade of C. gloeosporioides. To confirm the pathogenicity of the fungus, 20 tejocote fruits were surface disinfested by immersion in a 1% sodium hypochlorite solution for 1 min, washed three times with sterile distilled water and dried on sterilized filter paper. Inoculations were performed by deposition of 10 μl of a conidial suspension (106 spores ml-1) on the fruit surface. Ten fruit were mock inoculated with distilled water as a control. All fruits were kept in a moist chamber at 25°C for 10 days. Pathogenicity test was repeated twice. Disease symptoms were observed on all inoculated fruit after 7 days, whereas control fruit did not develop symptoms. Fungal colonies were re-isolated from all symptomatic fruits and were found to be morphologically identical to the original isolate inoculated on tejocote fruits, thus fulfilling Koch´s postulates. In Mexico, García-Alvarez (1976) reported Colletotrichum sp. on fruits of Crataegus mexicana, however, that report was not supported by morphological characterization nor pathogenicity tests. To our knowledge, this is the first report of C. gloeosporioides causing anthracnose of Crataegus gracilior in Mexico and worldwide.Fil: Nieto López, Edgar Humberto. University of Nebraska; Estados UnidosFil: Everhart, Sydney. University of Nebraska; Estados UnidosFil: Ayala Escobar, Victoria. Colegio de Postgraduados; MéxicoFil: Camacho Tapia, Moises. Universidad Autónoma Chapingo; MéxicoFil: Bernardi Lima, Nelson. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigaciones Agropecuarias. Instituto de Patología Vegetal; ArgentinaFil: Nieto Angel, Raúl. Universidad Autónoma Chapingo; MéxicoFil: Tovar Pedraza, Juan Manuel. Universidad Autónoma Chapingo; Méxic

Hyperpigmentation of hard palate induced by chloroquine therapy

Abstract The antimalarials are one of the most commonly prescribed drugs for conditions such as lupus erythematosus and rheumatoid arthritis, and the side effects, though infrequent, are well known. The antimalarial agent chloroquine diphosphate usually causes pigmentary changes in the oral mucosa characterized by a bluish-grey to black discolorations mainly in the hard palate. Considering only the hard palate hyperpigmentation caused by chloroquine, to the best of our knowledge, only 13 cases have been reported in the English language literature. We described an additional case of palate hyperpigmentation related to the chronic use of chloroquine diphosphate in a 60-year-old Mexican woman. Although the diagnosis is usually made based on medication history and clinical presentation, a biopsy specimen may be helpful to confirm the diagnosis. Clinicians must be aware of these drugs and their adverse effects in order to make the correct diagnosis and decide on the optimal treatment for the condition. Key words: Oral cavity, hard palate, hyperpigmentation, chloroquine, antimalarial

Influence of boundary conditions on oxygen distribution in an organ-on-a-chip platform

Organ-on-a-chip (OoC) platforms have revolutionized the drug development process by offering an effective alternative to animal models. These advanced microfluidic platforms mimic the organ functions at a microscale, and they can be produced at a large scale and at a lower price. Despite the variety of OoC models developed up to now, the combination of numerical simulations with experimental procedures has been of paramount importance in the development of more realistic and effective OoC devices. In addition, a better understanding of the physical phenomena happening in OoC can be obtained. In the present work, fluid flow numerical simulations were carried out in an OoC aiming to evaluate the influence of imposing different inlet velocities on the oxygen distribution along the device. This is of great importance to understand if the oxygen that reaches the cells is adequate for their culture. The results showed that for the geometry tested, with four organ models in parallel, by increasing the inlet velocity, the dissolved oxygen where cells are cultured also increases. This proves the importance of using numerical simulations for improving the performance of the experimental tests and optimizing the flow conditions.&nbsp

New insights on the roles of bacteria and protozoa in activated-sludge processes

Wastewater treatment is one of the most important biotechnological processes in the world. Nevertheless, the highly complex microbial communities in wastewater treatment plants (WWTP) are barely understood, remaining, most of the time, as a “black box”. Even though many microbiological methods are available to study the activated-sludge communities, many recent questions on the role of a significant number of microorganisms are kept unanswered. The current model and design concepts of activated-sludge consider bacteria as the sole active biomass. The activities of all other microbial community members (protozoa, metazoa, phages, etc.) are hidden in a simple decay process responsible for the reduction of active biomass and there is an assumed difficulty in establishing the way of how the interactions between the bacterial and the eukaryotic populations can affect the performance of the treatment system. PROTOFILWW project (PTDC/AMB/68393/2006) aimed at the study of 37 WWTP during 2 years, allowing for the identification of the little metazoa, protozoa and filamentous bacteria in 296 samples and enabling the correlation of these communities with physical-chemical and operational parameters. Furthermore, several studies on related subjects were and are being carried on to enlighten the roles of the different organisms in the depurating process, on how they interact with each other and on methods to identify certain filamentous and floc-forming bacteria with relevant functions in these artificial ecosystems. Simultaneously, a web-based platform was conceived to ensure the dissemination of knowledge and the communication of results between the project team and the WWTP technicians and managers

Uso de biocarbón en la estimulación de la microbiota del suelo y el manejo del carbón del maní (Thecaphora frezii)

Dada la importancia del maní (Arachis hypogaea) para la economía de la provincia de Córdoba, surge el interés por dotar de valor agregado a los residuos que genera este sector productivo. Actualmente, se han desarrollado unidades de generación de energía eléctrica que utilizan la biomasa de cáscara de maní. A pesar de la eficiencia del proceso, las cenizas (C) que son el subproducto que se genera, se acumulan en grandes volúmenes que no tiene destino concreto. Por otra parte, una alternativa es la producción de biocarbón (BC), material producido por la pirólisis de biomasa con la finalidad de ser utilizado como enmienda de suelos. Su aplicación ha registrado aumento de la fertilidad e intercambio catiónico en suelos pobres en materia orgánica, retención de agua y nutrientes, y mejora del pH en suelos ácidos. A su vez, ha demostrado ser promisorio en la supresión de enfermedades por diversos mecanismos, inducción de resistencia sistémica, aumento de la abundancia y/o actividad de microorganismos benéficos, efecto fungitóxico por sus componentes químicos y/o por su estructura porosa que provee un hábitat que propicia el incremento de la microbiota nativa. Dado que el cultivo de maní se vio afectado en la última década por el carbón del maní (Thecaphora frezii), siendo la enfermedad del rizoplano de mayor prevalencia e intensidad, una opción para prevenirla es actuar sobre el suelo, con la finalidad de inactivar al patógeno. En este sentido, la aplicación de C o BC obtenidos a través del tratamiento de cáscaras de maní, podrían ser estrategias promisorias en la reducción de la incidencia de carbón del maní potenciando las aplicaciones de estos subproductos. El objetivo fue evaluar el efecto de C y BC en la actividad microbiana edáfica, parámetros fisiológicos y de respuesta al carbón del maníInstituto de Patología VegetalFil: Serri, Dannae Lilia. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patología Vegetal; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Unidad de Fitopatología y Modelización Agrícola (UFYMA); Argentina.Fil: Guzzo, María Carla. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Fisiología y Recursos Genéticos Vegetales; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Unidad de Estudios Agropecuarios; ArgentinaFil: Bernanrdi Lima, Nelson. Consejo Nacional de Investigaciones Científicas y Técnicas. Unidad de Fitopatología y Modelización Agrícola (UFYMA); ArgentinaFil: Milesi Delaye, Luis Antonio. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Pergamino; ArgentinaFil: Andriulo, Adrian Enrique. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Pergamino. Pergamino, Buenos Aires; ArgentinaFil: Fermanelli, C. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Tecnológica Nacional. Facultad Regional Córdoba. Centro de Investigación y Tecnología Química; ArgentinaFil: Saux, C. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Tecnológica Nacional. Facultad Regional Córdoba. Centro de Investigación y Tecnología Química; ArgentinaFil: Dominchin, M.F. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Córdoba. Instituto Multidisciplinario de Biología Vegetal; ArgentinaFil: Meriles, J.M. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Córdoba. Instituto Multidisciplinario de Biología Vegetal; ArgentinaFil: Vargas Gil, Silvina. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patología Vegetal; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Unidad de Fitopatología y Modelización Agrícola (UFYMA); Argentin

An overview of the recent developments on fructooligosaccharide production and applications

Over the past years, many researchers have suggested that deficiencies in the diet can lead to disease states and that some diseases can be avoided through an adequate intake of relevant dietary components. Recently, a great interest in dietary modulation of the human gut has been registered. Prebiotics, such as fructooligosaccharides (FOS), play a key role in the improvement of gut microbiota balance and in individual health. FOS are generally used as components of functional foods, are generally regarded as safe (generally recognized as safe status—from the Food and Drug Administration, USA), and worth about 150€ per kilogram. Due to their nutrition- and health-relevant properties, such as moderate sweetness, low carcinogenicity, low calorimetric value, and low glycemic index, FOS have been increasingly used by the food industry. Conventionally, FOS are produced through a two-stage process that requires an enzyme production and purification step in order to proceed with the chemical reaction itself. Several studies have been conducted on the production of FOS, aiming its optimization toward the development of more efficient production processes and their potential as food ingredients. The improvement of FOS yield and productivity can be achieved by the use of different fermentative methods and different microbial sources of FOS producing enzymes and the optimization of nutritional and culture parameter; therefore, this review focuses on the latest progresses in FOS research such as its production, functional properties, and market data.Agencia de Inovacao (AdI)-Project BIOLIFE reference PRIME 03/347. Ana Dominguez acknowledges Fundacao para a Ciencia e a Tecnologia, Portugal, for her PhD grant reference SFRH/BD/23083/2005

5to. Congreso Internacional de Ciencia, Tecnología e Innovación para la Sociedad. Memoria académica

El V Congreso Internacional de Ciencia, Tecnología e Innovación para la Sociedad, CITIS 2019, realizado del 6 al 8 de febrero de 2019 y organizado por la Universidad Politécnica Salesiana, ofreció a la comunidad académica nacional e internacional una plataforma de comunicación unificada, dirigida a cubrir los problemas teóricos y prácticos de mayor impacto en la sociedad moderna desde la ingeniería. En esta edición, dedicada a los 25 años de vida de la UPS, los ejes temáticos estuvieron relacionados con la aplicación de la ciencia, el desarrollo tecnológico y la innovación en cinco pilares fundamentales de nuestra sociedad: la industria, la movilidad, la sostenibilidad ambiental, la información y las telecomunicaciones. El comité científico estuvo conformado formado por 48 investigadores procedentes de diez países: España, Reino Unido, Italia, Bélgica, México, Venezuela, Colombia, Brasil, Estados Unidos y Ecuador. Fueron recibidas un centenar de contribuciones, de las cuales 39 fueron aprobadas en forma de ponencias y 15 en formato poster. Estas contribuciones fueron presentadas de forma oral ante toda la comunidad académica que se dio cita en el Congreso, quienes desde el aula magna, el auditorio y la sala de usos múltiples de la Universidad Politécnica Salesiana, cumplieron respetuosamente la responsabilidad de representar a toda la sociedad en la revisión, aceptación y validación del conocimiento nuevo que fue presentado en cada exposición por los investigadores. Paralelo a las sesiones técnicas, el Congreso contó con espacios de presentación de posters científicos y cinco workshops en temáticas de vanguardia que cautivaron la atención de nuestros docentes y estudiantes. También en el marco del evento se impartieron un total de ocho conferencias magistrales en temas tan actuales como la gestión del conocimiento en la universidad-ecosistema, los retos y oportunidades de la industria 4.0, los avances de la investigación básica y aplicada en mecatrónica para el estudio de robots de nueva generación, la optimización en ingeniería con técnicas multi-objetivo, el desarrollo de las redes avanzadas en Latinoamérica y los mundos, la contaminación del aire debido al tránsito vehicular, el radón y los riesgos que representa este gas radiactivo para la salud humana, entre otros