Ciencia e Ingeniería en Alimentos y Biotecnología

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Subject: search.filters.subject.BACTERIÓFAGOS

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    Bioprospección de bacteriófagos líticos contra bacterias patógenas causantes de marchitez bacteriana en cultivos selectos de tomate riñón en Tungurahua. Articulado al proyecto de investigación aprobado mediante Resolución Nro. UTA-CONIN-2023-0294-R.
    (Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Biotecnología, 2025-02) Orozco Burbano, Erika Tatiana; Calero Cáceres, William Ricardo
    Bacterial wilt is one of the most serious plant diseases worldwide, caused by the presence of pathogenic bacteria in soil and plants. It mainly affects the Solanaceae family, including tomato crops, causing large economic losses due to the destruction of crops. In this study, bacteria present in crops with bacterial wilt symptoms of tomato crops in Tungurahua were isolated, identified and characterized. Biochemical and morphological tests were performed to rule out strains of no interest in the study. A MALDI-ToF analysis was then performed in order to identify the pathogen strains causing the disease. K. cowanii and C. flaccumfaciens were found to be the main plant pathogenic bacteria in the area. Additionally, 20 bacteriophages present in soil samples with different types of morphology were isolated and purified, only against K. cowanii, due to the absence of bacteriophages for C. flaccumfaciens. By performing the drop test with different bacterial hosts, it was shown that the isolated bacteriophages have a high specific lytic capacity for K. cowanii and do not present any risk against beneficial bacteria. Bioprospecting of the bacteriophages found suggests their use as a potential biological control agent against important phytopathogenic diseases such as bacterial wilt. The results obtained highlight the use of bacteriophages as a sustainable alternative benefiting the agricultural systems of the province of Tungurahua.
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    Bioprospección de bacteriófagos líticos contra bacterias patógenas causantes de marchitez bacteriana en cultivos selectos de tomate riñón en Tungurahua. Articulado al proyecto de investigación aprobado mediante Resolución Nro. UTA-CONIN-2023-0294-R.
    (Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Biotecnología, 2025-02) Altamirano Chérrez, Edgar Ricardo; Calero Cáceres William Ricardo
    Bacterial wilt is one of the most serious plant diseases worldwide, caused by the presence of pathogenic bacteria in soil and plants. It mainly affects the Solanaceae family, including tomato crops, causing large economic losses due to the destruction of crops. In this study, bacteria present in crops with bacterial wilt symptoms of tomato crops in Tungurahua were isolated, identified and characterized. Biochemical and morphological tests were performed to rule out strains of no interest in the study. A MALDI-ToF analysis was then performed in order to identify the pathogen strains causing the disease. K. cowanii and C. flaccumfaciens were found to be the main plant pathogenic bacteria in the area. Additionally, 20 bacteriophages present in soil samples with different types of morphology were isolated and purified, only against K. cowanii, due to the absence of bacteriophages for C. flaccumfaciens. By performing the drop test with different bacterial hosts, it was shown that the isolated bacteriophages have a high specific lytic capacity for K. cowanii and do not present any risk against beneficial bacteria. Bioprospecting of the bacteriophages found suggests their use as a potential biological control agent against important phytopathogenic diseases such as bacterial wilt. The results obtained highlight the use of bacteriophages as a sustainable alternative benefiting the agricultural systems of the province of Tungurahua.
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    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 Ricardo
    Bacteriophages 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.
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    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 Ricardo
    Food 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.
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    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 Ricardo
    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.
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    Bioprospección de bacteriófagos líticos de Salmonella enterica multirresistente de origen avícola de la provincia de Tungurahua
    (Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Biotecnología, 2023-09) Topa Pila, Paulina Liseth; Calero Cáceres, William Ricardo
    The use of bacteriophages has emerged as a promising alternative to address the antimicrobial resistance crisis, driven by the indiscriminate use of antibiotics in the poultry industry. This study isolated lytic bacteriophages specific to multi-resistant S. enterica of poultry origin in Tungurahua province from contaminated water sources and evaluated them based on their lytic profiles and low-temperature stability. Under the Microbiology and Biotechnology research line, the isolation and purification of bacteriophages was carried out using two strains of S. enterica serovar Infantis, by the double-layer agar method. Lytic profiles were evaluated by the gout test. The stability of the bacteriophages was tested through the reduction of the phage titre, at refrigeration and freezing temperatures, with pH 5.5 and 7.2. We isolated a total of 27 bacteriophages. Evaluation against 38 Salmonella isolates from poultry and five different S. enterica serovars revealed seven bacteriophages with a broad host range. Stability tests of bacteriophages F6, F7M1, and F4M1B, showed that temperature and pH significantly influenced phage titer reduction and bacterial lysis, underscoring the importance of understanding bacteriophages stability conditions for successful application. The results highlight the potential use of these lytic bacteriophages in designing biocontrol products for the poultry industry.
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    Bioprospección de bacteriófagos líticos de Salmonella enterica multirresistente de origen avícola de la provincia de Tungurahua
    (Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Biotecnología, 2023-09) Morales Cunalata, Katheryne Yolanda; Calero Cáceres, William Ricardo
    The use of bacteriophages has emerged as a promising alternative to address the antimicrobial resistance crisis, driven by the indiscriminate use of antibiotics in the poultry industry. This study isolated lytic bacteriophages specific to multi-resistant S. enterica of poultry origin in Tungurahua province from contaminated water sources and evaluated them based on their lytic profiles and low-temperature stability. Under the Microbiology and Biotechnology research line, the isolation and purification of bacteriophages was carried out using two strains of S. enterica serovar Infantis, by the double-layer agar method. Lytic profiles were evaluated by the gout test. The stability of the bacteriophages was tested through the reduction of the phage titre, at refrigeration and freezing temperatures, with pH 5.5 and 7.2. We isolated a total of 27 bacteriophages. Evaluation against 38 Salmonella isolates from poultry and five different S. enterica serovars revealed seven bacteriophages with a broad host range. Stability tests of bacteriophages F6, F7M1, and F4M1B, showed that temperature and pH significantly influenced phage titer reduction and bacterial lysis, underscoring the importance of understanding bacteriophages stability conditions for successful application. The results highlight the potential use of these lytic bacteriophages in designing biocontrol products for the poultry industry.
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    Prospección de bacteriófagos líticos de Salmonella en muestras ambientales de las provincias de Tungurahua y Cotopaxi, y su efecto sobre microorganismos patógenos
    (Universidad Técnica de Ambato. Facultad de Ciencia e Ingeniería en Alimentos y Biotecnología. Carrera de Ingeniería Bioquímica, 2020-08) Sailema Guevara, Vanessa Elizabeth; Calero Cáceres, William Ricardo
    According to the OMS, antimicrobial resistance (AMR) represents one of the main problems for public health worldwide. The genus Salmonella represents one of the most critical food pathogens, and its high prevalence in the Ecuadorian poultry sector poses a threat to public health. Considering the rising levels of AMR and the lack of new antibiotic molecules, the study of bacteriophages as potential tools against bacterial pathogens is regarded as an interesting alternative, due to their specificity and security. Twenty-six bacteriophages were isolated from water samples from the provinces of Tungurahua and Cotopaxi. The double-layer technique was applied. Subsequently, the lytic profiles of the bacteriophages against pathogenic strains were evaluated. Hierarchical clustering was performed to assess the similarity between lytic patterns. Two clusters were distinguished according to their lytic pattern. The inactivation kinetics of the most representative bacteriophages of each lysis cluster were evaluated. The phages f UTA 02 (S. Infantis), f UTA 19, and f UTA 20 (S. Enteritidis) showed an interesting lytic effect, causing an entire elimination of their host after 48 hours.