Ciencia e Ingeniería en Alimentos y Biotecnología
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Item Evaluación de la producción y conservación de bacteriófagos líticos de Salmonella enterica con medios de cultivo económicos y sustancias crioprotectoras alternativas(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Biotecnología, 2024-02) Romero Rovayo, Nathaly Johanna; Calero Cáceres, William RicardoBacteriophages represent an alternative for the elimination of potentially pathogenic microorganisms. However, their use involves numerous stages that include selection, generation of high phage titers, preservation, characterization, and assessments of their effectiveness. These processes require a significant financial investment in culture media and preservation methods, which could limit their applicability in scaling up. Therefore, in this study, cost-effective culture media were generated and alternative cryoprotective substances were used to improve the efficiency in obtaining and preserving high phage titers, thus facilitating their future industrial use. The formulation of culture media was carried out, using the avian-origin strain S. enterica serovar Infantis S11V23 as the host, and the bacteriophage Berlinvirus sp. F6_melonhead. Initially, the behavior of the bacteria and the phage against each formulation was analyzed. Finally, alternative cryoprotective substances were employed to evaluate the preservation of the titer over a period of 56 days at temperatures of 4, 20, and -70 degrees Celsius. The feasibility of obtaining bacterial and phage biomass using cost-effective culture media based on malt extract and brewer's yeast extract was proven, being effective for generating high phage titers. Furthermore, the stability of the bacteriophages at -70 Degrees Celsius was achieved using de-lactosed milk as a cryoprotective substance. The results highlight the possibility of scaling up bacteriophage production with limited resources and greater efficiency in generating bioproducts for the control of S. enterica useful for the poultry industry.Item Caracterización de la producción de metabolitos secundarios por microorganismos fotosintéticos(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Biotecnología, 2024-02) Lalaleo Camino, Christian Josué; García Solís, Mario DanielGlobal population growth has driven demand for food resources, causing a negative environmental impact through the expansion of agriculture and livestock farming. In response, photosynthetic microorganisms have been highlighted as an alternative source of bioactive compounds, such as proteins, carbohydrates, and pigments. This project evaluated the impact of carbon and nitrogen on the growth kinetics, production of proteins, carbohydrates, and pigments of the CLMR1 isolate, as well as its influence on cell morphology. Autotrophic and mixotrophic cultures were carried out, using carbon dioxide with two organic carbon sources (ethylene glycol and sodium acetate) and different concentrations of sodium nitrate (0, 0.625, 1.25 and 2.5) grams per liter. The most outstanding results were observed in the culture with 2.5 grams per liter of sodium nitrate and ethylene glycol, achieving the highest cell concentration (21,000,000 cells per milliliter), maximum amount of carbohydrates (1015.78 micrograms per milliliter) and the highest synthesis of chlorophyll. and carotenoids (0.274 and 0.207 micrograms per milliliter). Additionally, treatment with 2.5 grams per liter of sodium nitrate and sodium acetate produced the highest protein concentration (2.31 micrograms per milliliter). However, cultures without sodium nitrate showed the lowest synthesis of each metabolite and the minimum biomass production (200,000 cells per milliliter). Morphological changes were evident in the cells, highlighting the importance of optimal culture conditions for the synthesis of metabolites and their application in the sustainable production of high-value biomolecules.Item Análisis genómico de los bacteriófagos líticos f6 y f14 de Salmonella enterica Serovar Infantis(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Biotecnología, 2024-02) Gavilanes Tiban, Joselyn Micaela; Calero Cáceres, William RicardoFood contamination by Salmonella represents an emerging global public health crisis, with poultry farms being the principal reservoirs of this pathogen. The excessive use of antibiotics on these farms contributes to the increase in antibiotic resistance. In response to this situation, lytic bacteriophages emerge as a promising alternative for the biocontrol of Salmonella. However, it is essential to perform a detailed analysis of the complete genome of these bacteriophages before their implementation, in order to identify the presence of virulence and antibiotic resistance genes that could affect the safety of their in vivo implementation. In this study, two lytic bacteriophages of S. enterica serovar Infantis, named F6_melonhead and F14_guaytamphage, were sequenced using third-generation Oxford Nanopore technology. Genomic characterization revealed that F6 has a length of 29,097 bp and a GC content of 48.57 percent, encoding 30 functional proteins, while F14 has a length of 59,339 bp, a GC content of 56.44 percent, and encodes 49 functional proteins. In both bacteriophages, the presence of genes producing toxins, bacterial virulence factors, or antibiotic resistance genes was ruled out. Genetic comparison and phylogenetic analysis placed F6_melonhead and F14_guaytamphage within the Berlinvirus and Chivirus genera, respectively. These findings position F6_melonhead and F14_guaytamphage as promising candidates for the biocontrol of Salmonella in the poultry sector in Ecuador.Item Análisis genómico de los bacteriófagos líticos f6 y f14 de Salmonella enterica Serovar Infantis(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Biotecnología, 2024-02) Altamirano Cisneros, Karen Analía; Calero Cáceres, William Ricardowith poultry farms being the principal reservoirs of this pathogen. The excessive use of antibiotics on these farms contributes to the increase in antibiotic resistance. In response to this situation, lytic bacteriophages emerge as a promising alternative for the biocontrol of Salmonella. However, it is essential to perform a detailed analysis of the complete genome of these bacteriophages before their implementation, in order to identify the presence of virulence and antibiotic resistance genes that could affect the safety of their in vivo implementation. In this study, two lytic bacteriophages of S. enterica serovar Infantis, named F6_melonhead and F14_guaytamphage, were sequenced using third-generation Oxford Nanopore technology. Genomic characterization revealed that F6 has a length of 29,097 bp and a GC content of 48.57 percent, encoding 30 functional proteins, while F14 has a length of 59,339 bp, a GC content of 56.44 percent, and encodes 49 functional proteins. In both bacteriophages, the presence of genes producing toxins, bacterial virulence factors, or antibiotic resistance genes was ruled out. Genetic comparison and phylogenetic analysis placed F6_melonhead and F14_guaytamphage within the Berlinvirus and Chivirus genera, respectively. These findings position F6_melonhead and F14_guaytamphage as promising candidates for the biocontrol of Salmonella in the poultry sector in Ecuador.Item Producción de bioetanol utilizando biomasa de un cultivo mixto de microalgas como materia prima(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Biotecnología, 2022-09) Carrera Ocampo, Miguel Ángel; García Solís, Mario DanielThe biotechnological potential of microalgae in biofuels production continues to be a promising alternative to face the current energy crisis. This research project aimed to use the biomass of a mixed microalgae culture to produce bioethanol, for which the biomass production using industrial by-products as additional sources of carbon in the culture media was evaluated, and the optimal conditions for the acid hydrolysis of the carbohydrates contained in the microalgal biomass were determined. The mixotrophic cultures, composed of carbon dioxide and cheese whey or ethylene glycol as carbon sources, allowed the generation of higher biomass yields (1.38 and 0.60 milligrams per milliliter) in comparison with the autotrophic culture using carbon dioxide solely (0.50 milligrams per milliliter). Regarding the acid hydrolysis of carbohydrates, the higher release of reducing sugars per dry biomass (0.27 milligrams per milligram) was obtained using 2.0 percent volume-volume sulfuric acid. Additionally, it was determined that it is possible to obtain a higher amount of reducing sugars from the biomass obtained in the culture medium supplemented with ethylene glycol (0.30 milligrams per milligram). Finally, ethanol was produced by Saccharomyces cerevisiae at a concentration per dry biomass of 0.35 milligrams per milligram with a process yield of 82 percent from the fermentation of the sugars present in the biomass hydrolysate obtained from the culture with ethylene glycol. The results support the use of the biomass of the microalgae consortium as an efficient alternative to produce third-generation bioethanol.Item Estudio de factibilidad para la creación de una empresa de laboratorios virtuales para simulación de procesos microbiológicos y bioquímicos en la ciudad de Ambato, provincia de Tungurahua(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Ingeniería Bioquímica, 2021-09) Torres Paredes, Damián Raúl; Valencia Silva, Alex FabiánThe practical component within the academic and professional training of students allows linking the theory learnt in the classroom with reality. However, several limitations have been identified that have been accentuated by the COVID-19 health emergency. For this reason, the educational field has seen the need to implement technological tools that make it possible to meet the needs of education and modernise the teaching-learning process. Through the project developed, the feasibility was determined for the implementation of a virtual laboratory company for the simulation of microbiological and biochemical processes. The market study determined the acceptance of 25 and 26 percent of the sample of teachers and students respectively to acquire a licence for their own account. However, 100 percent of teachers and 66 percent of students agree that the educational institution should purchase these licences. The technical study located the company in the city of Ambato, province of Tungurahua, the technical characteristics of the product, the minimum requirements for its execution and the graphic identity criteria. Finally, an economic study and a financial analysis were carried out, where the project reflected an NPV value of 11342.76 USD, an IRR value of 38 percent, a benefit-cost ratio of 1.23 USD and a PRI value of 2 years and 7 months, thus demonstrating the viability of the project.Item Estudio de factibilidad para la creación de una empresa de laboratorios virtuales para simulación de procesos microbiológicos y bioquímicos en la ciudad de Ambato, provincia de Tungurahua(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Ingeniería Bioquímica, 2021-09) Moreno Iza, Karla Daniela; Valencia Silva, Alex FabiánThe practical component within the academic and professional training of students allows linking the theory learnt in the classroom with reality. However, several limitations have been identified that have been accentuated by the COVID-19 health emergency. For this reason, the educational field has seen the need to implement technological tools that make it possible to meet the needs of education and modernise the teaching-learning process. Through the project developed, the feasibility was determined for the implementation of a virtual laboratory company for the simulation of microbiological and biochemical processes. The market study determined the acceptance of 25 and 26 percent of the sample of teachers and students respectively to acquire a licence for their own account. However, 100 percent of teachers and 66 percent of students agree that the educational institution should purchase these licences. The technical study located the company in the city of Ambato, province of Tungurahua, the technical characteristics of the product, the minimum requirements for its execution and the graphic identity criteria. Finally, an economic study and a financial analysis were carried out, where the project reflected an NPV value of 11342.76 USD, an IRR value of 38 percent, a benefit-cost ratio of 1.23 USD and a PRI value of 2 years and 7 months, thus demonstrating the viability of the project.Item Determinación de la velocidad máxima de transferencia de oxígeno y la velocidad específica máxima de consumo de oxígeno en cultivos por lote de la bacteria (Bacillus subtilis DS23) en un biorreactor(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Ingeniería Bioquímica, 2020-01) Robalino Travez, Daniela Estefanía; Cerda Mejía, Liliana AlexandraThis research was carried out with the purpose of optimizing the production of biosurfactant with amphipathic activity from Bacillus subtilis DS23, which has been characterized as one of the microorganisms that have the best performance when producing this type of compounds. The experimentation was carried out in a 5-liter bioreactor maintaining constant parameters such as pH at 6.5, temperature at 30 Celsius degrees and aeration in 1 volume of air per volume of medium per minute (vvm), working with speeds of agitation of 150, 200 and 300 revolutions per minute (rpm) being this the only parameter that was varied. It was shown that the production of biomass and bio-surfactant in a stirred tank bioreactor is strongly influenced by agitation. The best results of both biomass and biosurfactant were obtained at 300 rpm, agitation speed that was used to determine the parameters proposed in this investigation. Using the Humphrey dynamic method using air as an oxygen-carrying gas, the effect of dissolved oxygen in the medium on the growth and production of biosurfactant was assessed by estimating the volumetric oxygen transfer coefficient (KLa), the specific rate of oxygen consumption (qO2) and the oxygen transfer rate (VTO) obtaining 0.002 h-1, 0.079 mmol O2 for L * h and 0.312 h-1 respectively. It was decided to evaluate these parameters because they are key in aerobic crops, and they are intended to serve as scaling criteria in future research.Item Diversidad biológica y metabólica de actinomicetes aislados de suelo y arena de ecosistemas glaciares de la Antártida y del norte del Chimborazo(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos. Carrera de Ingeniería Bioquímica, 2015) Villacís Barrazueta, Juan José; Rodríguez Meza, Carlos AlbertoItem Identificación de bacterias aisladas de ecosistemas glaciares, andinos y antárticos(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos. Carrera de Ingeniería Bioquímica, 2019-05) Palacios Mazón, Diego Marcelo; Rodríguez Meza, Carlos AlbertoBacteria isolated from glacial, Andean and Antarctic ecosystems were identified. By rep-PCR using the BOX primer, 72 strains were genotypically characterized and a dendrogram was constructed by numerical taxonomy, using the Pearson coefficient and the UPGMA algorithm, obtaining the formation of 10 groups. From these, representatives were selected for the sequencing of the 16S rDNA gene and phenotypic characterization. The phylogenetic analysis of the 16S rDNA gene was performed by calculating the evolutionary distances of Jukes & Cantor and the Neighbor-Joining algorithm, allowing the identification of 11 different species. Of which, strains DG-222 and DG-203 probably belong to new species. However, further studies are necessary to demonstrate this. Through the phenotypic characterization, a dendrogram of numerical taxonomy of phenotypic data was constructed using the SSM coefficient and the UPGMA algorithm, obtaining the formation of 21 phenetic groups with 84,6 percent similarity. Most of the collection was determined as Gram negative, psychrotroph, neutrotolerant, weak halophilic or halophilic. In addition, new physiological characteristics of 4 different species were discovered. The analysis of polyphasic taxonomy with the strains that had fingerprinting patterns similar to the identified species, managed to locate 35 strains according to the molecular characterization. Forming 16 phenetic groups at 90 percent similarity and determining the polyphase taxonomic position of 48,6 percent of the collection. Probably, the diversity of the collection is underestimated and there could be species that have not been identified in this study. It is necessary to carry out a second analysis of the strains that were excluded in the analyzes.
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