Evaluación de los efectos intestinales y cambios en la longevidad del nematodo Caenorhabditis elegans cepa mutante AX1410 y cepa silvestre N2 expuestas a la acción de Lactobacillus rhamnosus y Lactobacillus plantarum

Al ser una puerta de entrada de diferentes agentes biológicos, el sistema digestivo alberga gran cantidad de bacterias, correspondiente aproximadamente a un 90% de la flora general del intestino, considerándose uno de los lugares en donde estos microorganismos tienen mayor impacto; de hecho, se ha v...

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Autores:
Solano Meneses, Dayana
Marmolejo González, Everth David
Tipo de recurso:
Trabajo de grado de pregrado
Fecha de publicación:
2022
Institución:
Colegio Mayor de Cundinamarca
Repositorio:
Repositorio Colegio Mayor de Cundinamarca
Idioma:
spa
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Acceso en línea:
https://repositorio.unicolmayor.edu.co/handle/unicolmayor/5721
Palabra clave:
Probióticos
Modelo de estudio
Sistema digestivo
Herramientas terapéuticas
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Derechos Reservados - Universidad Colegio Mayor de Cundinamarca, 2022
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dc.title.spa.fl_str_mv Evaluación de los efectos intestinales y cambios en la longevidad del nematodo Caenorhabditis elegans cepa mutante AX1410 y cepa silvestre N2 expuestas a la acción de Lactobacillus rhamnosus y Lactobacillus plantarum
title Evaluación de los efectos intestinales y cambios en la longevidad del nematodo Caenorhabditis elegans cepa mutante AX1410 y cepa silvestre N2 expuestas a la acción de Lactobacillus rhamnosus y Lactobacillus plantarum
spellingShingle Evaluación de los efectos intestinales y cambios en la longevidad del nematodo Caenorhabditis elegans cepa mutante AX1410 y cepa silvestre N2 expuestas a la acción de Lactobacillus rhamnosus y Lactobacillus plantarum
Probióticos
Modelo de estudio
Sistema digestivo
Herramientas terapéuticas
title_short Evaluación de los efectos intestinales y cambios en la longevidad del nematodo Caenorhabditis elegans cepa mutante AX1410 y cepa silvestre N2 expuestas a la acción de Lactobacillus rhamnosus y Lactobacillus plantarum
title_full Evaluación de los efectos intestinales y cambios en la longevidad del nematodo Caenorhabditis elegans cepa mutante AX1410 y cepa silvestre N2 expuestas a la acción de Lactobacillus rhamnosus y Lactobacillus plantarum
title_fullStr Evaluación de los efectos intestinales y cambios en la longevidad del nematodo Caenorhabditis elegans cepa mutante AX1410 y cepa silvestre N2 expuestas a la acción de Lactobacillus rhamnosus y Lactobacillus plantarum
title_full_unstemmed Evaluación de los efectos intestinales y cambios en la longevidad del nematodo Caenorhabditis elegans cepa mutante AX1410 y cepa silvestre N2 expuestas a la acción de Lactobacillus rhamnosus y Lactobacillus plantarum
title_sort Evaluación de los efectos intestinales y cambios en la longevidad del nematodo Caenorhabditis elegans cepa mutante AX1410 y cepa silvestre N2 expuestas a la acción de Lactobacillus rhamnosus y Lactobacillus plantarum
dc.creator.fl_str_mv Solano Meneses, Dayana
Marmolejo González, Everth David
dc.contributor.advisor.none.fl_str_mv Gómez Jiménez, Martha
Sánchez Mora, Ruth Mélida
dc.contributor.author.none.fl_str_mv Solano Meneses, Dayana
Marmolejo González, Everth David
dc.subject.proposal.spa.fl_str_mv Probióticos
Modelo de estudio
Sistema digestivo
topic Probióticos
Modelo de estudio
Sistema digestivo
Herramientas terapéuticas
dc.subject.proposal.eng.fl_str_mv Herramientas terapéuticas
description Al ser una puerta de entrada de diferentes agentes biológicos, el sistema digestivo alberga gran cantidad de bacterias, correspondiente aproximadamente a un 90% de la flora general del intestino, considerándose uno de los lugares en donde estos microorganismos tienen mayor impacto; de hecho, se ha visto que dicha influencia tiene un efecto positivo en otros sistemas al intervenir en procesos metabólicos, de estimulación celular o protección a infecciones, por lo que en la actualidad esté sistema es considerado un blanco terapéutico prometedor para diversas enfermedades y garantizar su estabilidad es un reto para los investigadores. Con respecto a esto, los probióticos resultan ser una herramienta efectiva para el mantenimiento de este sistema, ya que contribuyen a la estabilización y recuperación de la flora intestinal, siendo Lactobacillus uno de los géneros con mayor cantidad de cepas implementadas. A pesar de esto y de su potencial como herramienta terapéutica, requieren de mayores estudios que esclarezcan los mecanismos por los cuales actúan, lo que puede realizarse por medio de la exposición de modelos biológicos de estudio a diferentes cepas de este género (1). Por lo anterior, en el presente estudio se evaluaron dos cepas de Lactobacillus, usando Caenorhabditis elegans como modelo de estudio por sus características morfológicas a nivel intestinal, ventajas en el costo de mantenimiento y facilidad de monitoreo, con el fin de ampliar los conocimientos ante el microorganismo probiótico y proponer al mismo como herramienta terapéutica ante patologías relacionadas con el eje intestinal
publishDate 2022
dc.date.accessioned.none.fl_str_mv 2022-10-20T20:08:27Z
dc.date.available.none.fl_str_mv 2022-10-20T20:08:27Z
dc.date.issued.none.fl_str_mv 2022
dc.type.spa.fl_str_mv Trabajo de grado - Pregrado
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dc.relation.references.spa.fl_str_mv Muñoz MJ. Longevity and heat stress regulation in Caenorhabditis elegans [Internet]2003;124(1):43-8. Available from: https://www.sciencedirect.com/science/article/abs/pii/S0047637402001689?via%3Dihub
FAO, OMS. Probióticos en los alimentos Propiedades saludables y nutricionales y directrices para la evaluación. Estudios FAO alimentación y nutrición [Internet]. 2006;85:52. Available from: file:///C:/Users/Acer/Documents/paty/homework1/PROBIOTICOS OPS 2006.pdf
Turnbaugh PJ, Ley RE, Hamady M, Fraser-Liggett CM, Knight R, Gordon JI. The Human Microbiome Project. Nature [Internet]. 2007;449(7164):804–10. Available from: https://www.nature.com/articles/nature06244#citeas
Kaveh Ashraf. Obesity and the regulation of fat metabolism. WormBook [Internet]. 2007;1–20. Available from: http://www.wormbook.org/chapters/www_obesity/obesity.pdf
Sáenz, Tomás Agurto RJ. BACTERIAS ÁCIDO LÁCTICAS : BIOPRESERVANTE DE LOS ALIMENTOS. 2008;8:54–64. Available from: file:///C:/Users/Admsoporte/Downloads/865-Texto del artículo-1883-1-10-20170902.pdf
Salvetti E, Torriani S, Felis GE. The Genus Lactobacillus: A Taxonomic Update. Probiotics and Antimicrobial Proteins [Internet]. 2012;4(4):217–26. Available from: https://ezproxy.unicolmayor.edu.co:2425/article/10.1007/s12602-012-9117-8
rompone G, Martorell P, Llopis S, González N, Genovés S, Mulet AP, et al. Anti-Inflammatory Lactobacillus rhamnosus CNCM I-3690 Strain Protects against Oxidative Stress and Increases Lifespan in Caenorhabditis elegans. PLoS ONE. 2012;7(12).
Kim Y, Mylonakis E. Caenorhabditis elegans immune conditioning with the probiotic bacterium Lactobacillus acidophilus strain ncfm enhances gram-positive immune responses. Infection and Immunity [Internet]. 2012;80(7):2500–8. Available from: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3416467/pdf/zii2500.pdf
Mackowiak PA. Recycling Metchnikoff: Probiotics, the intestinal microbiome and the quest for long life. Frontiers in Public Health. 2013;1(NOV):1–3.
Olveira G, González-molero I. Endocrinología y Nutrición Actualización de probióticos , prebióticos y simbióticos. 2016;63(9):482–94. Available from: https://www.elsevier.es/es-revista-endocrinologia-nutricion-12-articulo-actualizacion-probioticos-prebio ticos-simbioticos-nutricion-S1575092216301139
Nakagawa, Hisako et al. “Effects and mechanisms of prolongevity induced by Lactobacillus gasseri SBT2055 in Caenorhabditis elegans.” Aging cell vol. 15,2 (2016): 227-36. doi:10.1111/acel.12431
arrote A BR. Probioticos. Farmacia Abierta [Internet]. 2017;31(5):13–6. Available from: https://www.enfermeriaaps.com/portal/probioticos-farmacia-profesional-2017
Heeney D, Gareau M MM. Intestinal Lactobacillus in health and disease, a driver or just along for the ride? Physiol Behav [Internet]. 2017;176(12):139–48. Available from: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5808898/
Kamaladevi A, Balamurugan K. Lactobacillus casei triggers a TLR mediated RACK-1 dependent p38 MAPK pathway in Caenorhabditis elegans to resist Klebsiella pneumoniae infection. Food and Function [Internet]. 2016;7(7):3211–23. Available from: https://pubs.rsc.org/en/content/articlelanding/2016/FO/C6FO00510A
Azat R, Liu Y, Li W, Kayir A, Lin D bo, Zhou W wen, et al. Probiotic properties of lactic acid bacteria isolated from traditionally fermented Xinjiang cheese. Journal of Zhejiang University: Science B [Internet]. 2016;17(8):597–609. Available from: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4980438/pdf/JZUSB17-0597.pdf
Smolentseva O, Gusarov I, Gautier L, Shamovsky I, Defrancesco AS, Losick R, et al. Mechanism of biofilm-mediated stress resistance and lifespan extension in C. elegans. Scientific Reports [Internet]. 2017;7(1):1–16. Available from: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5540977/ %0A
Watts JL, Ristow M. Lipid and carbohydrate metabolism in Caenorhabditis elegans. Genetics [Internet]. 2017;207(2):413–46. Available from: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5629314/pdf/413.pdf
Yoon D, Lee M-H, Cha D. Measurement of Intracellular ROS in Caenorhabditis elegans Using 2’,7’-Dichlorodihydrofluorescein Diacetate. Bio-Protocol [Internet]. 2018;8(6):1–8. Available from: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5937271/
Plaza-Díaz J, Ruiz-Ojeda FJ, Gil-Campos M, Gil A. Immune-mediated mechanisms of action of probiotics and synbiotics in treating pediatric intestinal diseases. Nutrients [Internet]. 2018;10(1):1–20. Available from: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5793270/
Mantegazza C, Molinari P, D’Auria E, Sonnino M, Morelli L, Zuccotti GV. Probiotics and antibiotic-associated diarrhea in children: A review and new evidence on Lactobacillus rhamnosus GG during and after antibiotic treatment. Pharmacological Research [Internet]. 2018;128:63–72. Available from: http://dx.doi.org/10.1016/j.phrs.2017.08.001
Park HEH, Jung Y, Lee SJ v. Survival assays using Caenorhabditis elegans. Molecules and Cells [Internet]. 2018;40(2):90–9. Available from: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5339508/pdf/molce-40-2-90.pdf
Bhardwaj A, Thapliyal S, Dahiya Y, Babu K. FLP-18 functions through the G-protein-coupled receptors NPR-1 and NPR-4 to modulate reversal length in Caenorhabditis elegans. Journal of Neuroscience. 2018 May 16;38(20):4641–54.
Tankou SK, Regev K, Healy BC, Tjon E, Laghi L, Cox LM, et al. A probiotic modulates the microbiome and immunity in multiple sclerosis. Annals of Neurology [Internet]. 2018;83(6):1147–61. Available from: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6181139/
Liu YW, Liong MT, Tsai YC. New perspectives of Lactobacillus plantarum as a probiotic: The gut-heart-brain axis. Journal of Microbiology [Internet]. 2018;56(9):601–13. Available from: https://link.springer.com/article/10.1007/s12275-018-8079-2
Schnadower D, Tarr PI, Casper TC, Gorelick MH, Dean JM OK et al. Lactobacillus rhamnosus GG versus Placebo for Acute Gastroenteritis in Children. Physiol Behav [Internet]. 2017;176(12):139–48. Available from: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6358014/
Dimov I, Maduro MF. The C. elegans intestine: organogenesis, digestion, and physiology. Cell and Tissue Research [Internet]. 2019;377(3):383–96. Available from: https://ezproxy.unicolmayor.edu.co:2425/content/pdf/10.1007/s00441-019-03036-4.pdf
Meng Y, Wang J, Wang Z, Zhang G, Liu L, Huo G, et al. Lactobacillus plantarum KLDS1.0318 ameliorates impaired intestinal immunity and metabolic disorders in cyclophosphamide-treated mice. Frontiers in Microbiology [Internet]. 2019;10(APR). Available from: https://www.frontiersin.org/articles/10.3389/fmicb.2019.00731/ful
Zimmermann J, Obeng N, Yang W, Pees B, Petersen C, Waschina S, et al. The functional repertoire contained within the native microbiota of the model nematode Caenorhabditis elegans. ISME Journal [Internet]. 2020;14(1):26–38. Available from: http://dx.doi.org/10.1038/s41396-019-0504-y
Sharma K, Pooranachithra M, Balamurugan K, Goel G. Multivariate Analysis of Increase in Life Span of Caenorhabditis elegans Through Intestinal Colonization by Indigenous Probiotic Strains. Probiotics and Antimicrobial Proteins [Internet]. 2019;11(3):865–73. Available from: https://ezproxy.unicolmayor.edu.co:2425/content/pdf/10.1007/s12602-018-9420-0.pdf
Roselli M, Schifano E, Guantario B, Zinno P, Uccelletti D, Devirgiliis C. Caenorhabditis elegans and probiotics interactions from a prolongevity perspective. International Journal of Molecular Sciences [Internet]. 2019;20(20):1–14. Available from: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6834311/pdf/ijms-20-05020.pdf
Zeng L, Yang Z, Yun T, Fan S, Pei Z, Chen Z, et al. Antiaging effect of a Jianpi-yangwei formula in Caenorhabditis elegans. BMC Complementary and Alternative Medicine [Internet]. 2019;19(1):1–10. Available from: https://bmccomplementmedtherapies.biomedcentral.com/articles/10.1186/s12906-019-2704-4
Le TAN, Selvaraj B, Lee JW, Kang K. Measuring the effects of bacteria and chemicals on the intestinal permeability of caenorhabditis elegans. Journal of Visualized Experiments [Internet]. 2019;2019(154):1–10. Available from: https://www.jove.com/t/60419/measuring-effects-bacteria-chemicals-on-intestinal-permeability
Goya E , C, Ball KL, Stanley-wall NR, Xue F, Sampedro-torres-quevedo C, et al. Probiotic Bacillus subtilis Protects against a -Synuclein Aggregation in C . elegans Article Probiotic Bacillus subtilis Protects against a -Synuclein Aggregation in C . elegans. 2020;367–80. Available from: https://www.cell.com/action/showPdf?pii=S2211-1247%2819%2931743-7
Wieërs G, Belkhir L, Enaud R, Leclercq S, Philippart de Foy JM, Dequenne I, et al. How Probiotics Affect the Microbiota. Frontiers in Cellular and Infection Microbiology. 2020;9(January).
Poupet C, Chassard C, Nivoliez A, Bornes S. Caenorhabditis elegans, a Host to Investigate the Probiotic Properties of Beneficial Microorganisms. Frontiers in Nutrition [Internet]. 2020;7(August):1–22. Available from: https://www.frontiersin.org/articles/10.3389/fnut.2020.00135/full
Liu H, Li D, Zhang R, Sun L, Wang D. Lipid metabolic sensors of MDT-15 and SBP-1 regulated the response to simulated microgravity in the intestine of Caenorhabditis elegans. Biochemical and Biophysical Research Communications [Internet]. 2020;528(1):28–34. Available from: https://doi.org/10.1016/j.bbrc.2020.05.099
ydoun S, Choi H, Dela-Cruz G, Kruempel J, Huang S, Bazopoulou D, Miller H. An alternative food source for metabolism and longevity studies in Caenorhabditis elegans. Commun Biol 4, 258 (2021). https://doi.org/10.1038/s42003-021-01764-4
dc.rights.spa.fl_str_mv Derechos Reservados - Universidad Colegio Mayor de Cundinamarca, 2022
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spelling Gómez Jiménez, Marthaa140977a794b3ad529765cfe0ead6c4eSánchez Mora, Ruth Mélidaefe8f5fbfdf7ec7c24ba850634bfbb9aSolano Meneses, Dayanabc6f833ed8f9112370342e5e456dd6dbMarmolejo González, Everth David0d11913ecf454407e8c24ba57fb5956c2022-10-20T20:08:27Z2022-10-20T20:08:27Z2022https://repositorio.unicolmayor.edu.co/handle/unicolmayor/5721Al ser una puerta de entrada de diferentes agentes biológicos, el sistema digestivo alberga gran cantidad de bacterias, correspondiente aproximadamente a un 90% de la flora general del intestino, considerándose uno de los lugares en donde estos microorganismos tienen mayor impacto; de hecho, se ha visto que dicha influencia tiene un efecto positivo en otros sistemas al intervenir en procesos metabólicos, de estimulación celular o protección a infecciones, por lo que en la actualidad esté sistema es considerado un blanco terapéutico prometedor para diversas enfermedades y garantizar su estabilidad es un reto para los investigadores. Con respecto a esto, los probióticos resultan ser una herramienta efectiva para el mantenimiento de este sistema, ya que contribuyen a la estabilización y recuperación de la flora intestinal, siendo Lactobacillus uno de los géneros con mayor cantidad de cepas implementadas. A pesar de esto y de su potencial como herramienta terapéutica, requieren de mayores estudios que esclarezcan los mecanismos por los cuales actúan, lo que puede realizarse por medio de la exposición de modelos biológicos de estudio a diferentes cepas de este género (1). Por lo anterior, en el presente estudio se evaluaron dos cepas de Lactobacillus, usando Caenorhabditis elegans como modelo de estudio por sus características morfológicas a nivel intestinal, ventajas en el costo de mantenimiento y facilidad de monitoreo, con el fin de ampliar los conocimientos ante el microorganismo probiótico y proponer al mismo como herramienta terapéutica ante patologías relacionadas con el eje intestinalÍndice de figuras Índice de gráficas Índice de tablas RESUMEN INTRODUCCIÓN OBJETIVOS OBJETIVO GENERAL OBJETIVOS ESPECÍFICOS 1. ANTECEDENTES 2. MARCO REFERENCIAL 2.1. Probióticos 2.2. Selección de probióticos 2.3. Lactobacillus spp uno de los principales géneros probióticos 2.4. Viabilidad y probióticos 2.5. Modelos para el estudio de probióticos 2.5.1. Caenorhabditis elegans como modelo de estudio en diferentes enfermedades. 3. DISEÑO METODOLÓGICO 3.1. Cultivo y mantenimiento del nematodo 3.2. Cultivo y mantenimiento de bacterias probióticas 3.3. Sincronización del nematodo 3.4. Determinación de la concentración ideal del probiótico 3.5. Ensayo de preferencia alimentaria 3.6. Determinación de la vida útil del nematodo frente al tratamiento probiótico 3.7. Observación de cambios a nivel intestinal del nematodo frente al probiótico 3.8. Análisis estadístico 4. RESULTADOS 4.1. Identificación de los estadios larvarios de C. elegans 4.2. Concentración óptima de L. rhamnosus y L. plantarum incorporados en la dieta de C. elegans 4.3. Preferencia de C. elegans por el estado de la dieta 4.4. Longevidad de C. elegans tras el consumo de concentraciones seleccionadas del probiótico 4.5. Supervivencia al estrés térmico tras el consumo de L. rhamnosus y L. plantarum. 4.6. Evaluación cualitativa de la grasa intestinal en C. elegans tras el consumo de L. rhamnosus y L. plantarum. 5. DISCUSIÓN 6. CONCLUSIONES 7. REFERENCIAS BIBLIOGRÁFICAS 8. ANEXOSPregradoBacteriólogo(a) y Laboratorista Clínico46p.application/pdfspaDerechos Reservados - Universidad Colegio Mayor de Cundinamarca, 2022https://creativecommons.org/licenses/by-nc-sa/4.0/info:eu-repo/semantics/closedAccessAtribución-NoComercial-CompartirIgual 4.0 Internacional (CC BY-NC-SA 4.0)http://purl.org/coar/access_right/c_14cbEvaluación de los efectos intestinales y cambios en la longevidad del nematodo Caenorhabditis elegans cepa mutante AX1410 y cepa silvestre N2 expuestas a la acción de Lactobacillus rhamnosus y Lactobacillus plantarumTrabajo de grado - Pregradohttp://purl.org/coar/resource_type/c_7a1fhttp://purl.org/coar/version/c_970fb48d4fbd8a85Textinfo:eu-repo/semantics/bachelorThesishttps://purl.org/redcol/resource_type/TPinfo:eu-repo/semantics/publishedVersionUniversidad colegio mayor de CundinamarcaBacteriología y Laboratorio ClínicoMuñoz MJ. Longevity and heat stress regulation in Caenorhabditis elegans [Internet]2003;124(1):43-8. Available from: https://www.sciencedirect.com/science/article/abs/pii/S0047637402001689?via%3DihubFAO, OMS. Probióticos en los alimentos Propiedades saludables y nutricionales y directrices para la evaluación. Estudios FAO alimentación y nutrición [Internet]. 2006;85:52. Available from: file:///C:/Users/Acer/Documents/paty/homework1/PROBIOTICOS OPS 2006.pdfTurnbaugh PJ, Ley RE, Hamady M, Fraser-Liggett CM, Knight R, Gordon JI. The Human Microbiome Project. Nature [Internet]. 2007;449(7164):804–10. Available from: https://www.nature.com/articles/nature06244#citeasKaveh Ashraf. Obesity and the regulation of fat metabolism. WormBook [Internet]. 2007;1–20. Available from: http://www.wormbook.org/chapters/www_obesity/obesity.pdfSáenz, Tomás Agurto RJ. BACTERIAS ÁCIDO LÁCTICAS : BIOPRESERVANTE DE LOS ALIMENTOS. 2008;8:54–64. Available from: file:///C:/Users/Admsoporte/Downloads/865-Texto del artículo-1883-1-10-20170902.pdfSalvetti E, Torriani S, Felis GE. The Genus Lactobacillus: A Taxonomic Update. Probiotics and Antimicrobial Proteins [Internet]. 2012;4(4):217–26. Available from: https://ezproxy.unicolmayor.edu.co:2425/article/10.1007/s12602-012-9117-8rompone G, Martorell P, Llopis S, González N, Genovés S, Mulet AP, et al. Anti-Inflammatory Lactobacillus rhamnosus CNCM I-3690 Strain Protects against Oxidative Stress and Increases Lifespan in Caenorhabditis elegans. PLoS ONE. 2012;7(12).Kim Y, Mylonakis E. Caenorhabditis elegans immune conditioning with the probiotic bacterium Lactobacillus acidophilus strain ncfm enhances gram-positive immune responses. Infection and Immunity [Internet]. 2012;80(7):2500–8. Available from: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3416467/pdf/zii2500.pdfMackowiak PA. Recycling Metchnikoff: Probiotics, the intestinal microbiome and the quest for long life. Frontiers in Public Health. 2013;1(NOV):1–3.Olveira G, González-molero I. Endocrinología y Nutrición Actualización de probióticos , prebióticos y simbióticos. 2016;63(9):482–94. Available from: https://www.elsevier.es/es-revista-endocrinologia-nutricion-12-articulo-actualizacion-probioticos-prebio ticos-simbioticos-nutricion-S1575092216301139Nakagawa, Hisako et al. “Effects and mechanisms of prolongevity induced by Lactobacillus gasseri SBT2055 in Caenorhabditis elegans.” Aging cell vol. 15,2 (2016): 227-36. doi:10.1111/acel.12431arrote A BR. Probioticos. Farmacia Abierta [Internet]. 2017;31(5):13–6. Available from: https://www.enfermeriaaps.com/portal/probioticos-farmacia-profesional-2017Heeney D, Gareau M MM. Intestinal Lactobacillus in health and disease, a driver or just along for the ride? Physiol Behav [Internet]. 2017;176(12):139–48. Available from: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5808898/Kamaladevi A, Balamurugan K. Lactobacillus casei triggers a TLR mediated RACK-1 dependent p38 MAPK pathway in Caenorhabditis elegans to resist Klebsiella pneumoniae infection. Food and Function [Internet]. 2016;7(7):3211–23. Available from: https://pubs.rsc.org/en/content/articlelanding/2016/FO/C6FO00510AAzat R, Liu Y, Li W, Kayir A, Lin D bo, Zhou W wen, et al. Probiotic properties of lactic acid bacteria isolated from traditionally fermented Xinjiang cheese. Journal of Zhejiang University: Science B [Internet]. 2016;17(8):597–609. Available from: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4980438/pdf/JZUSB17-0597.pdfSmolentseva O, Gusarov I, Gautier L, Shamovsky I, Defrancesco AS, Losick R, et al. Mechanism of biofilm-mediated stress resistance and lifespan extension in C. elegans. Scientific Reports [Internet]. 2017;7(1):1–16. Available from: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5540977/ %0AWatts JL, Ristow M. Lipid and carbohydrate metabolism in Caenorhabditis elegans. Genetics [Internet]. 2017;207(2):413–46. Available from: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5629314/pdf/413.pdfYoon D, Lee M-H, Cha D. Measurement of Intracellular ROS in Caenorhabditis elegans Using 2’,7’-Dichlorodihydrofluorescein Diacetate. Bio-Protocol [Internet]. 2018;8(6):1–8. Available from: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5937271/Plaza-Díaz J, Ruiz-Ojeda FJ, Gil-Campos M, Gil A. Immune-mediated mechanisms of action of probiotics and synbiotics in treating pediatric intestinal diseases. Nutrients [Internet]. 2018;10(1):1–20. Available from: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5793270/Mantegazza C, Molinari P, D’Auria E, Sonnino M, Morelli L, Zuccotti GV. Probiotics and antibiotic-associated diarrhea in children: A review and new evidence on Lactobacillus rhamnosus GG during and after antibiotic treatment. Pharmacological Research [Internet]. 2018;128:63–72. Available from: http://dx.doi.org/10.1016/j.phrs.2017.08.001Park HEH, Jung Y, Lee SJ v. Survival assays using Caenorhabditis elegans. Molecules and Cells [Internet]. 2018;40(2):90–9. Available from: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5339508/pdf/molce-40-2-90.pdfBhardwaj A, Thapliyal S, Dahiya Y, Babu K. FLP-18 functions through the G-protein-coupled receptors NPR-1 and NPR-4 to modulate reversal length in Caenorhabditis elegans. Journal of Neuroscience. 2018 May 16;38(20):4641–54.Tankou SK, Regev K, Healy BC, Tjon E, Laghi L, Cox LM, et al. A probiotic modulates the microbiome and immunity in multiple sclerosis. Annals of Neurology [Internet]. 2018;83(6):1147–61. Available from: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6181139/Liu YW, Liong MT, Tsai YC. New perspectives of Lactobacillus plantarum as a probiotic: The gut-heart-brain axis. Journal of Microbiology [Internet]. 2018;56(9):601–13. Available from: https://link.springer.com/article/10.1007/s12275-018-8079-2Schnadower D, Tarr PI, Casper TC, Gorelick MH, Dean JM OK et al. Lactobacillus rhamnosus GG versus Placebo for Acute Gastroenteritis in Children. Physiol Behav [Internet]. 2017;176(12):139–48. Available from: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6358014/Dimov I, Maduro MF. The C. elegans intestine: organogenesis, digestion, and physiology. Cell and Tissue Research [Internet]. 2019;377(3):383–96. Available from: https://ezproxy.unicolmayor.edu.co:2425/content/pdf/10.1007/s00441-019-03036-4.pdfMeng Y, Wang J, Wang Z, Zhang G, Liu L, Huo G, et al. Lactobacillus plantarum KLDS1.0318 ameliorates impaired intestinal immunity and metabolic disorders in cyclophosphamide-treated mice. Frontiers in Microbiology [Internet]. 2019;10(APR). Available from: https://www.frontiersin.org/articles/10.3389/fmicb.2019.00731/fulZimmermann J, Obeng N, Yang W, Pees B, Petersen C, Waschina S, et al. The functional repertoire contained within the native microbiota of the model nematode Caenorhabditis elegans. ISME Journal [Internet]. 2020;14(1):26–38. Available from: http://dx.doi.org/10.1038/s41396-019-0504-ySharma K, Pooranachithra M, Balamurugan K, Goel G. Multivariate Analysis of Increase in Life Span of Caenorhabditis elegans Through Intestinal Colonization by Indigenous Probiotic Strains. Probiotics and Antimicrobial Proteins [Internet]. 2019;11(3):865–73. Available from: https://ezproxy.unicolmayor.edu.co:2425/content/pdf/10.1007/s12602-018-9420-0.pdfRoselli M, Schifano E, Guantario B, Zinno P, Uccelletti D, Devirgiliis C. Caenorhabditis elegans and probiotics interactions from a prolongevity perspective. International Journal of Molecular Sciences [Internet]. 2019;20(20):1–14. Available from: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6834311/pdf/ijms-20-05020.pdfZeng L, Yang Z, Yun T, Fan S, Pei Z, Chen Z, et al. Antiaging effect of a Jianpi-yangwei formula in Caenorhabditis elegans. BMC Complementary and Alternative Medicine [Internet]. 2019;19(1):1–10. Available from: https://bmccomplementmedtherapies.biomedcentral.com/articles/10.1186/s12906-019-2704-4Le TAN, Selvaraj B, Lee JW, Kang K. Measuring the effects of bacteria and chemicals on the intestinal permeability of caenorhabditis elegans. Journal of Visualized Experiments [Internet]. 2019;2019(154):1–10. Available from: https://www.jove.com/t/60419/measuring-effects-bacteria-chemicals-on-intestinal-permeabilityGoya E , C, Ball KL, Stanley-wall NR, Xue F, Sampedro-torres-quevedo C, et al. Probiotic Bacillus subtilis Protects against a -Synuclein Aggregation in C . elegans Article Probiotic Bacillus subtilis Protects against a -Synuclein Aggregation in C . elegans. 2020;367–80. Available from: https://www.cell.com/action/showPdf?pii=S2211-1247%2819%2931743-7Wieërs G, Belkhir L, Enaud R, Leclercq S, Philippart de Foy JM, Dequenne I, et al. How Probiotics Affect the Microbiota. Frontiers in Cellular and Infection Microbiology. 2020;9(January).Poupet C, Chassard C, Nivoliez A, Bornes S. Caenorhabditis elegans, a Host to Investigate the Probiotic Properties of Beneficial Microorganisms. Frontiers in Nutrition [Internet]. 2020;7(August):1–22. Available from: https://www.frontiersin.org/articles/10.3389/fnut.2020.00135/fullLiu H, Li D, Zhang R, Sun L, Wang D. Lipid metabolic sensors of MDT-15 and SBP-1 regulated the response to simulated microgravity in the intestine of Caenorhabditis elegans. Biochemical and Biophysical Research Communications [Internet]. 2020;528(1):28–34. Available from: https://doi.org/10.1016/j.bbrc.2020.05.099ydoun S, Choi H, Dela-Cruz G, Kruempel J, Huang S, Bazopoulou D, Miller H. An alternative food source for metabolism and longevity studies in Caenorhabditis elegans. 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Unicolmayorrepositorio@unicolmayor.edu.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