Ciencia e Ingeniería en Alimentos y Biotecnología

URI permanente para esta comunidadhttp://repositorio.uta.edu.ec/handle/123456789/412

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Determinación de los subtipos de Toxoplasma gondii en muestras de lechuga (Lactuca sativa l.) del mercado América de Ambato
(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Biotecnología, 2026-01)
Pacheco Jiménez, Yesenia Camila
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De La Torre Olvera, Helena Maritza
Toxoplasma gondii is a parasite that causes ocular toxoplasmosis, a zoonosis that affects one-third of the world's population and significantly harms pregnant women and immunocompromised individuals. It has been shown to have several routes of transmission, mainly related to cat feces and undercooked meat. However, there are also routes of transmission through raw vegetables, especially green leafy vegetables such as lettuce (Lactuca sativa L). In Ecuador, there is no information on the presence of this parasite in vegetable matrices, nor on its molecular characterization. The objective of this research was to determine the presence of T. gondii in lettuce sold in the América market in Ambato and to identify whether it belongs to subtype II or III by amplifying the SAG2 gene and using nested PCR. A descriptive and exploratory analysis was carried out, in which 21 samples from seven market stalls were analyzed. Eleven of these samples were identified microscopically, and seven of them showed molecular evidence of the presence of T. gondii (33, percent). However, none of the samples were compatible with subtypes II or III, suggesting that non-archetypal strains of T. gondii may be circulating in the city. In conclusion, the study determined the presence of T. gondii in lettuce samples and provides preliminary evidence on the genetic variability of the parasite, laying the groundwork for future research.
Análisis de la presencia de ooquistes de Toxoplasma gondii en coles verdes (Brassica oleracea var. Capitata) comercializados en el mercado central de Ambato, Ecuador
(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Biotecnología, 2025-07)
Ibarra Santos John Esteban
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De la Torre Olvera Helena Maritza
Toxoplasmosis caused by Toxoplasma gondii constitutes a critical public health problem affecting approximately one-third of the world's population, with severe impacts on pregnant women and immunocompromised individuals, being particularly relevant in Ecuador where seroprevalence rates of 70 percent are reported and there is limited research on parasitic contamination in vegetable products. The general objective was to analyze the presence of Toxoplasma gondii oocysts in green cabbages (Brassica oleracea var. capitata) commercialized in the Central Market of Ambato, Ecuador. The methodology employed completely randomized sampling of 15 cabbages from 5 sales stalls, microscopic analysis through flotation with Sheather's sucrose solution, and molecular confirmation by PCR targeting the B1 gene. The results demonstrated a prevalence of 53.33 percent (16 of 30 samples) of oocyst contamination, with significant variations between stalls (16.7 percent to 100 percent) and a statistically significant association (chi-square of 9.911, p less than 0.05) between sales stall and parasitic presence. It is concluded that there is considerable risk to local public health due to the high prevalence detected. The main contribution to the state of the art lies in constituting the first research that documents Toxoplasma gondii in vegetables commercialized in Ambato, providing fundamental epidemiological data for control and prevention strategies in food safety, and establishing a validated methodological protocol for parasitic detection in vegetable matrices within the Ecuadorian context.
Diseño y estandarización de un protocolo de diagnóstico molecular por RT-PCR para la genotipificación de las variantes de VPH predominantes en Ecuador (Genotipos:16, 18, 45, 58, 31)
(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Biotecnología, 2023-09)
Limachi Coello, Kevin Patricio
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Núñez Villacís, Lorena de los Ángeles
Human Papillomavirus (HPV) is the most common sexually transmitted pathogen worldwide. Nowadays, there are more than 200 HPV genotypes classified into low-risk (LR-HPV) and high-risk HPV strains (HR-HPV). The HR-HPV group is precursor of cervical cancer, which represents the fourth most deadly type of cancer worldwide and the second leading cause of death of women in Ecuador. Early and specific diagnosis leads to an improvement in the quality of life, low mortality rates and help to choose an efficient treatment. This study was based on the development of a Real Time PCR (RT-PCR) protocol for the genotyping of HR-HPV variants found in Ecuador (16, 18, 45, 58 and 31). Which were chosen through bibliographic analysis of clinical-population studies and research carried out in Ecuador in the last 8 years. A bioinformatic analysis of HPV genomes was developed, which result in a set of 10 primers with the capacity to identify the mentioned genotypes by SYBR Green technology. In addition to a thermal cycling protocol for RT-PCR amplification standardized experimentally in an in-house test for Laboratory of Medical Specialties Ochoa & Ochoa. Finally, we get a protocol able to recognize the HPV genotypes 16, 18, 45, 58 and 31 with a minimal synthetic DNA amount of 0.05 picograms per microliter. This study could be used as a genotyping prototype to be evaluated in human clinical samples.
Diseño y validación in silico de primers para la construcción de las variantes mutantes S121E, S121D, D186H y R280A de la enzima PETasa de Ideonella sakaiensis mediante tres diferentes métodos de mutagénesis dirigida al sitio
(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Ingeniería Bioquímica, 2022-03)
Herrera Aldaz, Bryan Alexander
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Cerda Mejía, Liliana Alexandra
The PETase enzyme from Ideonella sakaiensis (IsPETase) can be used to degrade PET, however, although it has been reported to date to have the highest enzymatic activity under normal conditions of all PET degrading enzymes, its low thermal stability limits its analysis and application. Therefore, by targeted mutagenesis, specific mutations can be introduced into the DNA to significantly increase its enzymatic activity. The purpose of this study was the design and in silico validation of primers for the construction of S121E, S121D, D186H and R280A mutant variants of IsPETase using three site-directed mutagenesis techniques: QuikChange, Q5 Site-Directed Mutagenesis and Phusion Site-Directed Mutagenesis. For the design of the primers, the parameters contained in the design guidelines for targeted mutagenesis methods were considered. The results showed that the 24 primers obtained meet the general criteria such as length, melting temperature, percentage of GC content and the position of the mutation in the primer. For the validation of the proposed primers, hairpin, autodimer and heterodimer analysis was performed with bioinformatics tools, the calculated values are within acceptable ranges. Finally, the specificity of the primers design was carried out by means of alignments, whose results indicate a high specificity for their respective targets. Likewise, an in silico PCR assay confirmed the specificity of the primers, which represent the expected amplification product, thus demonstrating the correct selection of the designed primers and the execution of the mutagenic PCR process.
Diseño y validación in silico de primers para la amplificación del Gen GSTM
(Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Ingeniería Bioquímica, 2022-03)
Guevara Guevara, Kevin Joel
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Galarza Galarza, Cristian Fernando
Lung cancer is a disease that affects a large part of the population, there are several reasons why it develops, one of them involves the GSTM1 gene, for this reason the possible relationship with cancer was consulted through the databases Malacards and Harmonizome, using the NM_000561.4 sequence for primer design. The region taken for the design of the primers comprised between exon 5 and exon 7, since in this there is a SNP related to lung cancer, a substitution of a C for a G in position 534, resulting in the GSTM1 A and GSTM1 B alleles containing a lysine and an asparagine respectively at position 172, a third GSTM1 0 allele called the null allele generated by a homozygous deletion, which leads to susceptibility to lung cancer. Hence the importance of design for both diagnostic and research use. The design was carried out in the Primer-Blast and Primer3Plus programs with default parameters and another design modifying them, mainly the concentrations of divalent cations and dNTPs, the rest of the configurations were modified according to the purpose of the primers. Once the set of primers was obtained, compatibility was analyzed using the MultiPLX 2.1 program and the melting curves were obtained using the uMELT software. Finally, the in silico validation was carried out with the Oligoanalyzer and Beacon designer free edition programs.

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