Ciencia e Ingeniería en Alimentos y Biotecnología

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

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2025 - 20262

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Tipo de acceso: Acceso abiertoTema: search.filters.subject.COL VERDE

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    Item type:Ítem,
    Optimización de un protocolo de extracción de ADN para la detección molecular de ooquistes de Toxoplasma gondii en coles verdes (Brassica oleracea var. Capitata) procedentes del mercado Central de Ambato
    (Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Biotecnología, 2026-07)
    Barreno Vargas, Carlos Alexander
    ;
    De La Torre Olvera, Helena Maritza
    Toxoplasmosis is a foodborne zoonosis of public health relevance, particularly in pregnant women, neonates, and immunocompromised individuals. Raw vegetables may act as vehicles for Toxoplasma gondii oocysts due to contact with contaminated water, soil, and handling practices. In this context, green cabbage marketed in Ambato represents a sanitary matrix of interest, since its compact leaf structure favors the retention of parasitic particles and hinders their removal through washing. This study was framed within the Microbiology and Biotechnology research line, linked to food safety, and was developed using an experimental approach. Green cabbage samples were analyzed through recovery with PBS 1X whit Tween® 80, concentration using the Ritchie technique, microscopic observation with Lugol and safranin staining, and molecular evaluation. To optimize DNA extraction, a 2 cubed factorial design was applied, considering thermal lysis, mechanical lysis, and proteinase K digestion. The recovered DNA was quantified by spectrophotometry, assessed using A260 out of 280 and A260 out of 230 ratios, and verified by conventional PCR and electrophoresis, amplifying a 196 bp fragment. The results showed structures compatible with oocysts in 8 of 21 samples using Lugol staining and in 6 samples using safranin staining. The treatment with 10 thermal shock cycles, 2 mechanical lysis cycles, and 20 microliter of proteinase K showed the best overall performance. The scientific novelty lies in proposing a combined, cost-efficient protocol adapted to plant matrices, achieving 100 percentage PCR detection compared with 50 percentage using the GeneJET® kit.
  • Miniatura
    Item type:Ítem,
    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
    ;
    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.