Potential of New Bacterial Strains for a Multiproduct Bioprocess Application: A Case Study Using Isolates of Lactic Acid Bacteria from Pineapple Silage of Costa Rican Agro-Industrial Residues

Lactic acid bacteria (LAB) with potential for the development of multi-product processes are necessary for the valorization of side streams obtained during the biotechnological production of lactic acid (LA). In this study, 14 LAB strains isolated from pineapple agro-industrial residues in Costa Rica were cultivated in microplates, and the six strains with the highest growth were selected for fermentation in microbioreactors to evaluate the production of LA and acetic acid, and the consumption of glucose. Lacticaseibacillus paracasei 6710 and L. paracasei 6714 presented the highest OD600 values (1.600 and 1.602, respectively); however, the highest LA (in g/L) production was observed in L. paracasei 6714 (14.50 ± 0.20) and 6712 (14.67 ± 0.42). L. paracasei 6714 was selected for bioreactor fermentation and reached a maximum OD600 of 6.3062 ± 0.141, with a LA yield of 84.9% and a productivity of 1.06 g L−1 h −1 after 21 h of fermentation. Finally, lipoteichoic acid (LTA) detection from biomass was performed and the antimicrobial activity of the compounds present in the supernatant was studied. LTA was detected from L. paracasei 6714 biomass, and its supernatant caused significant inhibition of foodborne surrogate microorganisms. LAB isolated from pineapple silage have biotechnological potential for multiproduct processes.Universidad de Costa RicaUCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias de la Salud::Centro de Investigación en Enfermedades Tropicales (CIET)UCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias Agroalimentarias::Centro Nacional de Ciencia y Tecnología de Alimentos (CITA)UCR::Vicerrectoría de Docencia::Ciencias Agroalimentarias::Facultad de Ciencias Agroalimentarias::Escuela de Tecnología de AlimentosUCR::Vicerrectoría de Docencia::Salud::Facultad de Microbiologí

Potential of New Bacterial Strains for a Multiproduct Bioprocess Application: A Case Study Using Isolates of Lactic Acid Bacteria from Pineapple Silage of Costa Rican Agro-Industrial Residues

Lactic acid bacteria (LAB) with potential for the development of multi-product processes are necessary for the valorization of side streams obtained during the biotechnological production of lactic acid (LA). In this study, 14 LAB strains isolated from pineapple agro-industrial residues in Costa Rica were cultivated in microplates, and the six strains with the highest growth were selected for fermentation in microbioreactors to evaluate the production of LA and acetic acid, and the consumption of glucose. Lacticaseibacillus paracasei 6710 and L. paracasei 6714 presented the highest OD600 values (1.600 and 1.602, respectively); however, the highest LA (in g/L) production was observed in L. paracasei 6714 (14.50 ± 0.20) and 6712 (14.67 ± 0.42). L. paracasei 6714 was selected for bioreactor fermentation and reached a maximum OD600 of 6.3062 ± 0.141, with a LA yield of 84.9% and a productivity of 1.06 g L−1 h−1 after 21 h of fermentation. Finally, lipoteichoic acid (LTA) detection from biomass was performed and the antimicrobial activity of the compounds present in the supernatant was studied. LTA was detected from L. paracasei 6714 biomass, and its supernatant caused significant inhibition of foodborne surrogate microorganisms. LAB isolated from pineapple silage have biotechnological potential for multiproduct processes

Assessment of Different Lactic Acid Bacteria Isolated from Agro-Industrial Residues: First Report of the Potential Role of Weissella soli for Lactic Acid Production from Milk Whey

Abstract: The production of lactic acid (LA) through the microbial conversion of agro-industrial residuals is an important process in the biotechnology industry. The growth kinetics of 30 strains of lactic acid bacteria (LAB) isolated from agro-industrial residues were determined and nine strains were selected for microbioreactor fermentation. Lactiplantibacillus pentosus_70-1 (1.662) and L. pentosus_ 19-2 (1.563) showed the highest OD600 values, whereas the highest growth rates were observed for L. pentosus_19-2 (0.267 h−1) and Weissella soli_31 (0.256 h−1). The production of LA and acetic acid (AA), glucose consumption, and metabolic profiles were determined, without finding significant differences in the production of LA; however, W. soli_29 produced the highest amount of LA (20.833 gL−1) and was able to metabolize most of the studied carbohydrates. Based on these results, W. soli_29 was chosen for a 20 h fermentation in a 7 L bioreactor using both standard medium and milk whey supplemented medium. W. soli_29 produced 16.27 gL−1 and 7.21 gL−1 of LA in each of these mediums, respectively. These results show the underlying potential of Weissella strains for biotechnological applications. Additional analysis which should contemplate different agro-industrial residues and other conditions in bioreactors must be carried out.UCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias de la Salud::Centro de Investigación en Enfermedades Tropicales (CIET)UCR::Vicerrectoría de Docencia::Salud::Facultad de MicrobiologíaUCR::Vicerrectoría de Docencia::Ingeniería::Facultad de Ingeniería::Escuela de Ingeniería de BiosistemasUCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias Agroalimentarias::Centro Nacional de Ciencia y Tecnología de Alimentos (CITA

First Molecular Evidence of Tomato chlorotic spot virus Infecting Tomatoes in Cuba

UCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias Básicas::Centro de Investigación en Biología Celular y Molecular (CIBCM)UCR::Vicerrectoría de Docencia::Ciencias Agroalimentarias::Facultad de Ciencias Agroalimentarias::Escuela de Tecnología de Alimento

First Report of Tomato yellow leaf curl virus in Tomato in Costa Rica

One of the most important invasive and harmful members of the genus Begomovirus (family Geminiviridae) is the monopartite Tomato yellow leaf curl virus (TYLCV), which is widespread over the world associated with tomato yellow leaf curl disease (TYLCD). Tomato (Solanum lycopersicum) plants infected with TYLCV show upward leaf curling and yellowing. In Latin America, isolates of TYLCV have been reported from Cuba, the Dominican Republic, Mexico and Puerto Rico (1), Guatemala (GenBank Accession No. GU355941), and Venezuela (partial genome sequence DQ302033). In Costa Rica, only isolates of the bipartite begomoviruses Tomato leaf curl Sinaloa virus (TLCSiV) (3) and Tomato yellow mottle virus (KC176780, KC176781) have been reported infecting tomatoes. During a survey conducted in 2012, similar begomovirus-like symptoms (leaf yellowing and upward leaf curling) were observed in tomato plants of five commercial growing areas in the Central Valley (Grecia region) of Costa Rica. In total, 65 tomato samples were randomly collected, 14 from greenhouses and 41 from open fields. Symptoms of upward leaf curling and yellowing were observed in three samples. Total DNA was extracted from collected samples and tested by dot blot hybridization using a probe to the coat protein (CP) gene of a Guatemalan isolate of Bean golden yellow mosaic virus (3). Only the three symptomatic samples tested positive, which represents an incidence of 14% (2 samples) in greenhouses and 2.4% (1 sample) in open field crops. These samples were subjected to rolling circle amplification (RCA) for viral circular genome amplification (2). The amplified products were then digested with MspI restriction endonuclease, which resulted into DNA fragments of 2,320 and 458 bp for all three samples. This suggested infection with a monopartite begomovirus. In order to obtain the full-length clone, the RCA product of two samples (5240 and 5241) was digested with BamHI, and the ~2.8 kb DNA fragment was cloned into pBluescript II SK(+) (Stratagene, La Jolla, CA) vector. After transformation of Escherichia coli DH5α, recombinant plasmids with inserts of expected size were selected and the insert was sequenced by primer walking (Macrogen Inc., Korea). The inserts of three clones from the two samples (CR:5240-16:2012, CR:5240-17:2012, and CR:5241-14:2012) were sequenced (deposited in GenBank as KF533855, KF533856, and KF533857, respectively). Sequences were all 2,781 nt long and shared 100% identity between themselves (1-nt mismatch between CR:5240-16:2012 and CR:5240-17:2012, and CR:5240-16:2012 and CR:5241-14:2012; and 2-nt mismatches between CR:5240-17:2012 and CR:5241-14:2012) and 99% with the sequence of Tomato yellow leaf curl virus-Israel[Japan:Haruno:2005] (TYLCV-IL[JR:Har:05]) (AB192966). These sequences represented full length genomes of isolates of the monopartite begomovirus TYLCV-IL and grouped in a phylogenetic clade (4) that comprised TYLCV-IL isolates reported from Asia (China and Japan) and from Mexico, while more distantly related to the clade comprising TYLCV-IL isolates reported from Central America (Cuba, Guatemala, Puerto Rico) and the United States, suggesting a distinct introduction event in Costa Rica. This is the first report of the presence of TYLCV in Costa Rica, therefore it is imperative to study the incidence and geographical spread of this virus in the country as well as its genetic diversity, since TYLCV infections might lead to significant yield losses, as reported in other countries.UCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias Básicas::Centro de Investigación en Biología Celular y Molecular (CIBCM

Multilocus typing sequences in Lactobacillus casei isolates from pineapple peels silages

Introducción. Lactobacillus casei se caracteriza por adaptarse al medio durante el proceso fermentativo. Objetivo. Caracterizar con tipificación de secuencias multilocus (MTLS), aislamientos de L. casei presentes en ensilados de cáscara de piña con niveles crecientes de urea. Materiales y métodos. Durante 2017 y con un diseño irrestricto al azar se prepararon veinte silos (2 kg). Cada cinco bolsas, se adicionaron 0, 0,5, 1 y 1,5 % de urea (p/p en base fresca). Después de treinta días, se tomó una muestra de cada repetición para realizar análisis bromatológicos. Se analizó materia seca (MS), proteína cruda (PC), carbohidratos no fibrosos, fibra en detergente ácido (FDA), fibra en detergente neutro (FDN), digestibilidad in vitro de la materia seca (DIVMS), hemicelulosa, extracto etéreo (EE), cenizas, nitrógeno amoniacal y pH. Se determinó el recuento de bacterias ácido-lácticas (BAL) para cada uno de los tratamientos. La identificación de las BAL se realizó con la secuencia del ARNr 16S. Los aislamientos de L. casei fueron analizados con MTLS. Resultados. Los cuatro tratamientos evaluados no presentaron diferencias significativas en la MS (12,10 a 12,86%), FDA (31,44 a 32,18 %), FDN (58,46 a 58,56 %), hemicelulosa (26,36 a 27,02 %), EE (2,45 a 2,96 %), cenizas (4,96 a 5,12%), nitrógeno amoniacal (0,45 a 0,49 %) y pH (3,36 a 3,48). No así, en PC (6,94 a 9,12 %) y DIVMS (82,84 a 84,72 %). La adición de urea se asoció a cambios en las poblaciones de BAL (6,56 a 6,90 UFC). Siete aislamientos de L. casei se identificaron y tipificaron con cinco genes distintos. Se encontraron entre dos (polA) y cinco (nrdD, pgm, mutL) alelos; esto permitió identificar un total de seis secuencias tipo (ST) distintas en los aislados de Costa Rica. Conclusión. Los aislamientos de L. casei se encontraron emparentados a bacterias del tracto gastrointestinal en humanos.Introduction. Lactobacillus casei is characterized by adapting to the environment during the fermentation process. Objective. To characterize with multilocus typing sequences (MTLS) isolates of L. casei present in pineapple peel silage with increasing levels of urea. Materials and methods. During 2017 and using an unrestricted random design, twenty silos (2 kg) were prepared. Every five bags, 0, 0.5, 1 and 1.5 % urea (w/w on fresh basis) were added. After thirty days, a sample was taken from each repetition for bromatological analysis. Dry matter (DM), crude protein (PC), non-fibrous carbohydrates, fiber in acid detergent (ADF), fiber in neutral detergent (NDF), in vitro digestibility of dry matter (IVDDM), hemicellulose, ethereal extract (EE), ashes, ammonia nitrogen, and pH. The Lactic Acid Bacteria (LAB) count was determined for each one of the treatments. Identification of LABs was carried out using the 16S rRNA sequence. The isolates of L. casei were analyzed using MTLS. Results. The four evaluated treatments did not present significant differences in DM (12.10 to 12.86 %), ADF (31.44 to 32.18 %), NDF (58.46 to 58.56 %), hemicellulose (26.36 to 27.02 %), EE (2.45 to 2.96 %), ash (4.96 to 5.12 %), ammonia nitrogen (0.45 to 0.49 %) and pH (3.36 at 3.48). Not so, in CP (6.94 to 9.12 %), and IVDDMS (82.84 to 84.72 %). The addition of urea was associated with changes in the BAL populations (6.56 to 6.90 CFU). Seven isolates of L. casei were identified and typed with five different genes. Between two (polA) and five (nrdD, pgm, mutL) alleles were found. This allowed the identification of a total of six different type sequences (ST) in the Costa Rican isolates. Conclusion. The L. casei isolates were found to be related to bacteria from the gastrointestinal tract in humans.Universidad de Costa Rica/[801-B5-505]/UCR/Costa RicaUCR::Vicerrectoría de Docencia::Ciencias Agroalimentarias::Facultad de Ciencias Agroalimentarias::Escuela de ZootecniaUCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias Agroalimentarias::Centro de Investigación en Nutrición Animal (CINA)UCR::Vicerrectoría de Docencia::Salud::Facultad de MicrobiologíaUCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias de la Salud::Centro de Investigación en Enfermedades Tropicales (CIET)UCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias Agroalimentarias::Centro Nacional de Ciencia y Tecnología de Alimentos (CITA)UCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias Agroalimentarias::Centro de Investigaciones Agronómicas (CIA)UCR::Vicerrectoría de Docencia::Ciencias Agroalimentarias::Facultad de Ciencias Agroalimentarias::Escuela de Tecnología de AlimentosUCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias Básicas::Centro de Investigación en Biología Celular y Molecular (CIBCM

Newly Discovered Natural Hosts of Tomato chlorosis virus in Costa Rica

Tomato chlorosis virus (ToCV) is an emerging whitefly-transmitted crinivirus. In Costa Rica in 2007, ToCV was detected in field-grown and greenhouse tomato (Solanum lycopersicum L.) plants causing symptoms of severe yellowing and foliar chlorosis.UCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias Básicas::Centro de Investigación en Biología Celular y Molecular (CIBCM

Effect of Gamma Irradiation and Selection with Fungus Filtrate (Rhizoctonia solani Kuhn) on the in Vitro Culture of Common Bean (Phaseolus vulgaris)

The present investigation was undertaken to study the effect of gamma irradiation (dose from 10 to 100 Gy) and in vitro selection with fungus filtrate as selecting agent (concentration from 20% to 100%) on the susceptibility of the common bean to Rhizoctonia solani. The best results were found with a dose of 20 Gy or a concentration of 20% of fungus filtrate applied separately. These conditions were used to evaluate the combined effect of both approaches in a second experiment. The combined effect of irradiation and then selection adversely affected growth (height and roots) and survival of the in vitro plants. It may not be necessary to combine the variation generated by irradiation with the selection technique. For future assays we propose the application of: 1) gamma radiation, thereby inducing not only mutants with pathogen resistance, but also with other agronomic traits of interest. Later in the subculture MV4 potential fungus-resistant mutants will be evaluated in the field; or 2) selection pressure using fungus filtrate during three subcultures, which may be sufficient to induce the variation necessary to obtain in vitro plants resistant to fungus.International Atomic Energy Agency/[COS5028]/IAEA/AustriaUniversidad de Costa Rica/[111-A8-210]/UCR/Costa RicaUCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias Básicas::Centro de Investigación en Biología Celular y Molecular (CIBCM)UCR::Vicerrectoría de Docencia::Ciencias Básicas::Facultad de Ciencias::Escuela de BiologíaUCR::Vicerrectoría de Docencia::Ciencias Sociales::Facultad de Ciencias Económicas::Escuela de Estadístic

Molecular characterization and genetic diversity of Jatropha curcas L. in Costa Rica

We estimated the genetic diversity of 50 Jatropha curcas samples from the Costa Rican germplasm bank using 18 EST-SSR, one G-SSR and nrDNA-ITS markers. We also evaluated the phylogenetic relationships among samples using nuclear ribosomal ITS markers. Non-toxicity was evaluated using G-SSRs and SCARs markers. A Neighbor-Joining (NJ) tree and a Maximum Likelihood (ML) tree were constructed using SSR markers and ITS sequences, respectively. Heterozygosity was moderate (He = 0.346), but considerable compared to worldwide values for J. curcas. The PIC (PIC = 0.274) and inbreeding coefficient (f = − 0.102) were both low. Clustering was not related to the geographical origin of accessions. International accessions clustered independently of collection sites, suggesting a lack of genetic structure, probably due to the wide distribution of this crop and ample gene flow. Molecular markers identified only one non-toxic accession (JCCR-24) from Mexico. This work is part of a countrywide effort to characterize the genetic diversity of the Jatropha curcas germplasm bank in Costa Rica.Consejo Nacional de Rectores/[736-B1-660]/CONARE/Costa RicaUCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias Básicas::Centro de Investigación en Biología Celular y Molecular (CIBCM

Análisis de secuencias polimórficas del núcleo y cloroplasto por conformación de ADN simple banda en introducciones de Cucurbita moschata Duchesne procedente de Mesoamérica

Tesis (magíster scientiae en biología con énfasis en genética y biología molecular)--Universidad de Costa Rica. Sistema de Estudios de Posgrado, 2007.UCR::Vicerrectoría de Investigación::Sistema de Estudios de Posgrado::Ciencias Básicas::Maestría Académica en Biologí