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:

• Know the theoretical bases of new culture-independent microbiological methodologies (DGGE, construction and analysis of gene libraries, FISH, metagenomics, etc.).
• Know the procedures required and recognize the steps of the protocols.
• Know the equipment, materials and reagents necessary for the different techniques.
• Recognize the application of different techniques and methods to use, if necessary, the most appropriate one for a sample or an ecosystem.
• Know the advantages and limitations of the different techniques and in which cases the use of each of them is appropriate.
• 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

Module II: Isolation and amplification of nucleic acids

4. Isolation and purification of nucleic acids.
5. PCR amplification
6. Quantitative-PCR or real-time PCR

Module III: Electrophoretic techniques

7. Denaturing Gradient Gel Electrophoresis (DGGE)
8. Other electrophoretic techniques (SSCP, LH-PCR, TFLP, RISA, ARISA, etc.).

Module IV: Massive sequencing techniques

9. Construction and analysis of gene libraries (gene libraries)
10. Metataxonomic and metagenomics techniques

Module V: Hybridization of nucleic acids

11. Hybridization. FISH hybridization. Microarray construction and hybridization

Module VI. Non-nucleic acid-based culture-independent techniques

12. Analysis of proteins and fatty acid esters. Spectroscopic techniques.