The course is aimed at industry professionals as well as health professionals dedicated to food microbiology. It is also aimed at students in the final years of their science degrees (Food Science and Technology, Human Nutrition and Dietetics, Microbiology, Biology, Biochemistry, Biotechnology, Pharmacy, Medicine, Veterinary Medicine) as well as master's and master's students. doctorate and the senior technicians of Professional Training in Dietetics, Oral Hygiene, Clinical and Medical Laboratory and Environmental Health. To all those who want to become familiar with a series of techniques, some of recent development, that are used to characterize the diversity, structure and succession of microbial populations in various ecosystems, including those of food matrices. These populations may have great technological or food security interest.

The syllabus has been prepared from the research experience of the professors/researchers who have developed the topics and the teaching experience that comes with having taken the course in person during three calls (2007, 2009 and 2011) before its online delivery. We hope that it is simple, enjoyable and, above all, useful in your work and/or research.

If you are interested in this training and want to know all the details about its content and operation, you can do so by sending an email to the course coordinators (Ana M. Garcia, Diego A. Moreno)

goals

In many ecological niches an unknown number of microbial species interact and compete for space and nutrients. Cells are found in diverse physiological and viability conditions, such that classical culture techniques underestimate diversity and are often not able to quantify even the majority groups. Difficulties in cultivating a specific microbial type can be due to various causes: need for unknown growth factors, dependency relationship with other microbes in the environment, being in non-cultivable physiological states, etc. To alleviate the limitations of cultivation, culture-independent molecular microbiological techniques have been developed with which the microorganisms of a given ecosystem are detected and/or quantified, depending on the techniques. In recent years these methods have been widely applied in many fields of Microbiology, including Food Microbiology. One of its main applications in this field is the study and characterization of traditional food fermentations. In this way, the components of the microbiota that direct the processes are identified, the population dynamics are studied throughout the production and maturation processes, and the key genes for the colonization of ecosystems and the maturation of products are tracked. . The learning objectives of the course are:

• Conocer las bases teóricas de las nuevas metodologías microbiológicas independientes de cultivo (DGGE, construcción y análisis de genotecas, FISH, metagenómica, etc.).
• Saber los procedimientos que se requieren y reconocer los pasos de los protocolos.
• Conocer el equipamiento, los materiales y los reactivos necesarios de las distintas técnicas.
• Reconocer la aplicación de las distintas técnicas y métodos para utilizar, llegado el caso, la más apropiada para una muestra o un ecosistema.
• Conocer las ventajas y limitaciones de las distintas técnicas y en qué casos resulta adecuada la utilización de cada una de ellas.
• Acquire the necessary knowledge to carry out a critical reading of the results reported in scientific articles with these techniques.

Agenda

The course program consists of twelve teaching units grouped into six modules where independent culture techniques in Food Microbiology are reviewed with a brief review of conventional analysis techniques and independent culture techniques not based on nucleic acids.

Module I: The Microbiology of Foods

1. Introduction to Food Microbiology
2. Microbiological analysis of foods: conventional techniques
3. Introduction to culture-independent microbiological techniques

Módulo II: Aislamiento y amplificación de ácidos nucleicos

4. Aislamiento y purificación de ácidos nucleicos.
5. Amplificación por PCR
6. PCR-cuantitativa o PCR a tiempo real

Módulo III: Técnicas electroforéticas

7. Electroforesis en geles de gradiente desnaturalizantes (DGGE)
8. Otras técnicas electroforéticas (SSCP, LH-PCR, TFLP, RISA, ARISA, etc.).

Módulo IV: Técnicas de secuenciación masiva

9. Construcción y análisis de librerías génicas (genotecas)
10. Técnicas metataxonómicas y metagenómicas

Módulo V: Hibridación de ácidos nucleicos

11. Hibridación. Hibridación FISH. Construcción e hibridación de microarrays

Módulo VI. Técnicas independientes de cultivo no basadas en ácidos nucleicos

12. Análisis de proteínas y ésteres de ácidos grasos. Técnicas espectroscópicas.