Ateneo - UPO

Module I, Part "Food Biotechnology"
Food biotechnology: introduction and significance; characteristics of microorganisms useful in biotechnology, the cell as a "molecular factory"; fermentations of interest to the food industry; beneficial microorganisms and the concept of "microbial starters"; prebiotics, probiotics, and synbiotics; fermented foods; industrial fermentations: classification and characteristics; fermentation plants: bioreactors and downstream processing, fermentation control parameters; fermentation reaction kinetics; enzyme and cell immobilization; biomass and Single Cell Proteins (SCP); examples of primary and secondary metabolites of food interest obtained via fermentation or recombinant technology: alcohols, sweeteners, neutral polysaccharides, organic acids, amino acids, flavorings, pigments; examples of bioconversions and bioremediation in the food sector (overview); enzymes: significance, production, and applications; second-generation food biotechnology: recombinant DNA, MOGMs (Microorganisms Obtained through Genetic Modification) and GMOs; relevant national and international regulations (overview); overview of analytical biotechnology; useful web resources in the field of biotechnology.
Module I, Part "Quality Control with Laboratory":
Analytical methods required to determine parameters listed on food product nutritional labels. Official analytical methods for regulated food matrices such as: drinking water, wine, milk, olive oil, milled grain products, and pasta. Instrumental analysis applied to foods, such as electrophoresis, GC-MS, HPLC-MS, and isotope ratio analysis. Laboratory exercises involving the analysis of wine, honey, fermented milk, and oil. Module II (Quality and Safety of Biotechnological Medicinal Products)
This module aims to provide fundamental knowledge regarding the quality principles of biological and biotechnological medicinal products, highlighting the role of the manufacturing process in defining product characteristics and performance in terms of quality, safety, and efficacy. Key technological strategies used to design, establish, and maintain the quality of biological and biotechnological drugs throughout their lifecycle will be examined in depth, with particular focus on process variability management and innovation in manufacturing technologies.
Module III (Applied Microbiology)
Microorganisms of industrial interest and their biotechnological applications
Principles of microbiological quality control in pharmaceutical manufacturing in compliance with current regulations; principles of microbiological control in food products, with particular focus on biofilm formation and control in production environments. Laboratory sessions featuring practical activities in isolation, enumeration, identification, and microbiological monitoring.

Slides used in the course, provided by the Teachers.
Module I “Biotecnologie Alimentari”
“BIOTECNOLOGIE ALIMENTARI”, Gigliotti-Verga; Piccin Editore Industrial microbiology and bioproductions: CHIMICA DELLE FERMENTAZIONI E MICROBIOLOGIA INDUSTRIALE, Marzona, II Edizione. PICCIN Editore. Applied Biotechnology: BIOTECNOLOGIE DI BASE, Colin Ratledge, Bjørn Kristiansen; Zanichelli Editore Deep insights in biotechnology: BIOTECNOLOGIA MOLECOLARE, Glick, Pasternak; Zanichelli Editore. Quality Control with laboratory: ANALISI DEI PRODOTTI ALIMENTARI, Cabras, Tuberoso, Ed. Piccin, Padova, 2014. For laboratory activities, handouts will be provided by the teacher.

Module II (Quality and Safety of Biotechnological Medicinal Products)
Crommelin D.J.A., Sindelar R.D., Meibohm B. Pharmaceutical Biotechnology – Fundamentals and Applications. Springer.
Course materials, selected scientific articles, and case studies provided by the instructor.

Module III (Applied Microbiology)
- Handbook of microbiological quality control: pharmaceuticals and medical devices Edited by Rosamund M. Baird, Norman A. Hodges, Stephen P. Denyer. - New York: CRC Press, c2000.-XVI, 254 p.
- Industrial Microbiology: An Introduction Edited by Michael J. Waites, Neil L. Morgan, John S. Rockey, Gary Higton; John Wiley & Sons, 01 apr 2009-304 p.
- Brock Biology of Microorganisms, Global Edition. Pearson - Biotecnologie microbiche. Stefano Donadio. Casa Editrice Ambrosiana
- Microbiologia industriale. Matilde Manzoni. Casa Editrice Ambrosiana
- Microbiologia farmaceutica. Nicola Carlone. Casa Editrice EdiSES
For laboratory activities, handouts will be provided by the teacher.

The course complements the student's knowledge in the fields of food science and pharmaceutical science, providing effective tools for the critical assessment and management of issues related to the production, quality, and safety of food, nutraceutical, and pharmaceutical products, including those of biotechnological origin.
The section on food biotechnology aims to provide students with fundamental knowledge in this field (principles of molecular biology; beneficial microorganisms and microbial starters; fermentation and precision fermentation; bioproduction and recovery of bioproducts of interest to the food, nutraceutical, and pharmaceutical sectors; biotechnological production of flavors and additives; enzymes and their food applications; recombinant biotechnology, GM microorganisms, and GMOs; relevant regulations). This knowledge will enable the management of quality and safety for products of biotechnological origin and provide the technical-scientific foundation for the biotechnological production of high-value-added molecules, ingredients, and materials for the food and pharmaceutical sectors, including upcycling techniques.
The section on quality control, which includes laboratory work, enables students to acquire fundamental knowledge of food analysis, helping them understand the significance of these analyses and their scope of application. Laboratory exercises guide students in applying their knowledge to the analysis of simple and complex food matrices, including in the context of food fraud detection. Students must demonstrate the ability to correctly determine and interpret analytical data and to communicate the results using appropriate methods.

The module II provides the fundamental knowledge required to understand the principles of quality of biotechnological medicinal products from the perspective of pharmaceutical technology. The course explores the role of the manufacturing process in building product quality, examining how process design, process variability control, and modern manufacturing strategies contribute to ensuring the quality, safety, and efficacy of biopharmaceuticals throughout their entire lifecycle.

The module III Applied Microbiology aims to provide students with the knowledge and methodological tools needed to understand the microbiological characteristics and application potential of the main microorganisms of biotechnological and industrial interest; acquire the principles and methods of microbiological quality control for pharmaceutical products, both sterile and non-sterile, according to current regulations; understand the principles of microbiological control of food and recognize the main foodborne pathogens; understand the phenomenon of biofilm formation in pharmaceutical and food production environments and the related control strategies; acquire practical laboratory skills in microbial isolation, enumeration, and identification, environmental monitoring, and the conduct of a microbial fermentation.

We recommend the acquisition of knowledge about the courses of Organic Chemistry, Biochemistry, Food chemistry and Microbiology fundamental for the understanding of the lessons of this course. The basic knowledge in Quantitative Drug Analysis laboratory (Analisi dei Farmaci I and II) is welcomed for laboratory experiences. Students are also expected to have a solid background in Pharmaceutical Technology and Molecular Biology, including the fundamentals of drug development and manufacturing, recombinant protein production, the structure and function of biological macromolecules, and the basic principles of pharmaceutical quality.
The compulsory prerequisites for the exams and the courses of Applied biochemisty and Food chemistry.

The teaching method involves in-person lectures focused on the discussion of specific case studies. Specialized seminars may be organized in collaboration with faculty members and industry experts, alongside "flipped classroom" sessions on specific topics; these aim to foster critical thinking among students, encourage active and responsible participation, and develop digital skills, autonomy, and collaborative abilities in group work.
For the Quality Control module—which includes a laboratory component—lectures are complemented by laboratory activities. Students are required to perform analyses independently by following and interpreting the provided protocols, documenting the steps taken in a laboratory notebook, performing the necessary calculations to express results in the correct units of measurement, and completing analysis reports. Instructions will also be provided on how to draft a comprehensive report regarding the complex food product analyzed. All course materials will be made available to students via the DIR platform.

The course, in its various forms, is to be understood as a unified program designed to round out the knowledge in the pharmaceutical-food sector required for professionals to manage issues related to the pharmaceutical-food and nutraceutical fields—covering both quality/safety control (analysis) and the biotechnological production of bioactive ingredients and foods. Laboratory activities involving the analysis of food matrices will take place in January; Microbiology lab will take place during the second semester.
Students with disabilities, Specific Learning Disorders (SLD), or Special Educational Needs (SEN) may request specific services and tools by contacting the Student Career Development and Coordination Staff and consulting the dedicated page on the University website: https://uniupo.it/it/servizi/servizi-studenti-disabili-e-dsa. Once they have contacted the University staff, students with disabilities, SLD, or SEN may also contact the course instructor to discuss the specific arrangements for exams and other educational aspects.

Student assessment is based on an oral exam covering all course topics, aimed at evaluating students' knowledge and critical skills—with particular reference to the role of the production process, variability management, and the technological strategies employed in the development and industrial production of biopharmaceuticals and "bioactive" ingredients for foods and dietary supplements.
Students must prepare a short PowerPoint presentation (maximum 4-5 slides) introducing an original in-depth topic of their choice—one not covered during the course but linked to one or more course themes through an interdisciplinary approach—and present it to the examination board. The discussion with the instructors will start from the presented topic and extend to all disciplines involved, integrating scientific information from the fields of pharmaceutical technology, pharmaceutical-food science, and applied microbiology.
Please note that for the second part of Module I (Quality Control with Laboratory), assessment includes a mandatory mid-term test—a prerequisite for laboratory access—covering all topics addressed in the preparatory lectures (including safety regulations). This test is structured as a written exam consisting of 7 multiple-choice questions, 2 calculation exercises, and 6 open-ended questions; notably, two of the open-ended questions require students to select the most appropriate analytical methods from those studied to solve a specific analytical problem. The mid-term test grade contributes to the final module assessment; if the result is unsatisfactory, it will be supplemented by oral questions during the laboratory period or at the end of the course via a specific oral exam session. The oral component typically involves three questions: a description of instrumental analysis methods, a description of an analysis performed in the laboratory (with particular emphasis on safety regulations), and a question regarding the selection of analytical methods. Evaluation will focus on both knowledge of the topics covered and the ability to critically discuss the course's core themes from an interdisciplinary perspective. Laboratory activities will also be assessed based on submitted analysis reports, the accuracy of calculations, and the completeness of the laboratory notebook. Finally, the ability to present the analytical data obtained from the complex matrix in a written report will be evaluated.
The overall grade calculation will take into account the varying university credit values of the individual modules. A minimum level of knowledge (a passing grade) for each module is required to pass the exam and earn the associated credits.

Module I, Part I (M. Arlorio): History/significance of food biotechnology. Molecules/macromolecules of interest. Prokaryotes vs. eukaryotes; transcription, translation, gene regulation (notes). The cell as a molecular factory. Alcoholic, homo- and hetero-lactic, malo-lactic, propionic, acetic, butyric and isobutyric, and citric fermentation. Precision fermentation. Virtuous microorganisms/starters. Cultivated fungi. Prebiotics, probiotics, synbiotics, post-biotics. Milk bioactives. Bacteriocins. Cyanobacteria and microalgae. Fermented foods: microbiology, fermentation aspects (milk/dairy products; alcoholic beverages; fermented vegetables and cured meats; leavened baked goods). Industrial fermentations; bioreactors. Fermentation monitoring. Mass transfer. Kinetics, fermentation yields. Enzyme/cell immobilization. Downstream processing. Biomass, Single Cell Proteins (SCP): red yeast rice, Pruteen, Quorn. Production of alcohols, amino acids, organic acids, neutral polysaccharides, vitamins, sweeteners (aspartame, glucose/maltose/fructose syrups, isomalt). Flavors: taste/odor receptors and perception; lactones, terpenes, vanillin; cyclodextrins. Carotenoid pigments. Industrial enzymes, classification, isolation, and production; applications (glucose syrups, milk delactosation, extraction technologies, production of chitosan and prebiotics, clarification and stabilization of juices/alcoholic beverages, accelerated cheese ripening, acrylamide reduction). Recombinant DNA. GMOs: production of r-chym and r-BGH. Manipulation and processing techniques in the plant field. GMOs: Flavr-Savr™ tomatoes, Bt™ corn, Round-up Ready™ soybeans, Golden Rice. Animal biotechnology (cloning, salmonids, mammals, other applications). National/international regulations. Biotechnology in food analytics (outline). Web resources.
Module I, Part II (M. Bordiga): Meaning and skills. Water: heating methods, Marcusson and Karl Fischer. Proteins: total nitrogen (Kjeldahl): principles, equipment, safety standards, correction factors. Protein extraction, SDS-PAGE electrophoresis, one- and two-dimensional urea-PAGE; isoelectrofocusing. Lipids: Soxhlet extraction, Gerber method for lipids in milk and dairy products. Refractive index, saponification number, iodine number. Lipids and quality: acidity, peroxide value, and p-anisidine value. Olive oils: regulations, panel test, spectrophotometric analysis, esterified fatty acids (GC-FID), characterization of unsaponifiable matter. Milk: specific gravity, freezing index, dry matter, acidity, peroxidase and phosphatase, soluble whey proteins. Cheese analysis. Sugars: reducing agents with the Fehling method, sucrose with polarimeter (Clerget), refractometry, degrees Brix. Fiber: composition, crude fiber, dietary fiber (total, soluble, and insoluble). Cereals and cereal products (flour, bread, and pasta): moisture, ash, gluten, acidity; bleaching agents and ascorbic acid. Wine: alcoholic strength, dry extract, total, fixed, and volatile acidity, proline, polyphenols, NMR and mass spectroscopy methods for assessing the origin of sugars. Drinking water: regulations for drinking and mineral water. Determination of ammonia (Nessler and phenol-hypochlorite), nitrites (Griess), nitrates. Oxidation number, BOD, COD. Hardness, chlorides, chlorine, sulfides, iron, chromium. Additives: Determination of benzoic acid in sugary beverages. Enzymatic analysis: meaning, sample preparation, methods, and protocols. Antioxidant activity of foods. Instrumental analyses: Applications of HPLC techniques with SPE extraction and purification; applications of GC techniques, aroma extraction with SPME. Atomic absorption: principles, applications. Laboratory: Basic analyses of wine, fermented milk, honey, and olive oil. Enzymatic determination of lactose, sucrose, fructose, and glucose. Determination of moisture, pH, ash, proteins, and acidity. Lipids, acidity, and peroxide values. Polarimetry. Protein analysis: extraction, SDS-PAGE. Polyphenols: Folin and DMAC.

Module II (M.L. Torre)
Distinction between biotechnological drugs and drugs obtained by chemical synthesis (structural and manufacturing characteristics). This module introduces the concept of quality as applied to biopharmaceuticals, highlighting the crucial role of the manufacturing process in defining product characteristics and the principle that "the process is the product."
The factors influencing the quality of biotechnological drugs will be analyzed, with particular reference to biological and technological variability, the relationship between the manufacturing process and product properties, and the impact of these aspects on the quality, safety, and efficacy of the drug.
The course will explore pharmaceutical development strategies aimed at designing and maintaining the quality of biopharmaceuticals, illustrating the principles of manufacturing process control, monitoring of critical process parameters, and modern technologies used to ensure the robustness and reproducibility of industrial production.
The effects of changes introduced in the manufacturing process during the life cycle of a biotechnological medicinal product, the principles of product comparability, and the scientific rationale behind the development of biosimilar medicinal products will also be discussed.
Finally, the module will present the main technological innovations in the production of biopharmaceuticals, with particular reference to continuous biomanufacturing, the digitalization of manufacturing processes, and the prospects of Quality 4.0, through the analysis and discussion of case studies from industrial practice.

Module III (L. Fracchia)
1. Microorganisms of biotechnological and industrial interest: Bioprospecting and microbial biodiversity for biotechnological, pharmaceutical, and industrial applications. Microorganisms: microbiological characteristics and applications; kinetics of microbiological processes: growth curve, batch, fed-batch, and continuous processes; methods for assessing microbial growth; microbial biosurfactants: production, characterization, and applications.
2. Microbiological quality control in the pharmaceutical and biotechnological fields: Microorganisms as biotechnological tools in pharmaceutical production; Main methods of sterilization of pharmaceutical products and biological indicators of sterilization; Sterility tests for bacteria/fungi; validation of sterility tests. Microbial contamination in non-sterile products: total microbial count, most probable number (MPN), filtration; Detection of specific microorganisms according to the European Pharmacopoeia; LAL (Limulus Amebocyte Lysate) test and determination of bacterial endotoxins; Good Manufacturing Practices (GMP) and environmental microbiological monitoring; Biofilms in pharmaceutical production environments: training, detection, and control strategies
3. Microbiological control in the food sector: Food pathogens and sources of contamination; Microbiological analysis methods; Biofilms on food processing surfaces: comparison with the pharmaceutical context
4. Laboratory activities: Isolation, enumeration, and identification of microorganisms from food samples (probiotic dairy product); Search for specific microorganisms (culture methods on selective/differential media); Environmental microbiological monitoring: air/surface sampling; Setup and monitoring of a microbial fermentation on a renewable agri-food matrix.

Gender mainstreaming: Teaching approaches will be used to ensure equal benefits for women and men in actions and activities and to eliminate gender inequalities, including through practical examples and selected case studies.

Students must demonstrate acquisition of the basic, specific, and advanced knowledge covered in the course and related to the application of first- and second-generation biotechnologies to food, nutraceutical, and pharmaceutical production models, with a particular focus on microbial and fermentation production.
They must demonstrate linguistic fluency and satisfactory technical-scientific presentation skills, demonstrating the ability to integrate the various modules and demonstrate interdisciplinarity. Students must demonstrate knowledge of the relevant regulatory framework and mastery of the most appropriate technique for managing an applied work situation (at the laboratory and industrial level, in the latter case as part of an interdisciplinary technical-scientific team). Specific expertise will be acquired in the field of genetically modified microorganisms/organisms, as well as their derived products for use in both the pharmaceutical and food sectors, as well as the underlying European regulatory status. Finally, students must demonstrate their ability to plan a biotechnology project applied to the pharmaceutical, food, and nutraceutical sectors.
For the "Quality Control with Laboratory" component of Module I, they must acquire basic and specific knowledge of the analytical methods applicable to the analysis of foods and dietary supplements, as well as the theoretical principles of instrumental analytical methods and their specific fields of application in the food sector (including product integrity and fraud prevention). They must acquire appropriate scientific language, demonstrate their ability to identify the most suitable analytical methods for solving a specific problem, and manage the information provided. During laboratory activities, students must be able to understand and correctly apply the provided analysis protocols, obtain the relevant analytical data, solve the calculations required to obtain the data in the correct unit of measurement, interpret the data in relation to any legal provisions specific to the matrix being examined, and independently make a final judgment.
By the end of Module II, students will have acquired the ability to understand the role of the manufacturing process in defining the characteristics of biological medicinal products relevant to quality, safety, and therapeutic efficacy. They will also be able to critically interpret the main sources of variability that characterize the industrial production of such medicinal products and discuss the technological strategies adopted to design, monitor, and control the manufacturing process, in order to ensure the consistency of the medicinal product's characteristics throughout its life cycle.
By the end of Module III, students will have acquired knowledge of the microbiological characteristics and biotechnological applications of the main microorganisms of industrial interest, the principles of microbiological quality control of pharmaceutical and food products, and the phenomenon of biofilm formation in production environments. They will also acquire the ability to apply the main microbiological techniques used in the pharmaceutical and food sectors, both theoretically and through practical laboratory work. At the end of the module III, students will be expected to demonstrate independent judgment in assessing the risk of microbial contamination and identifying the most appropriate prevention and control strategies. They will also have acquired technical and scientific communication skills appropriate for the comparison between the pharmaceutical and food sectors.