LEARNING OUTCOMES

I The course aims to broaden the knowledge on the most current biosecurity monitoring systems increasingly used in research, but also in the control of the circulation of goods, products marketed online, adulteration and fraud.

AIMS AND LEARNING OUTCOMES

The course aims to broaden the knowledge on the most current biosecurity monitoring systems increasingly used in research, but also in the control of the circulation of goods, products marketed online, sophistication and fraud.

In particular, topics such as:

• Biosecurity: General and introductory concepts;

• DNA profiling and its uses;

• Chemistry and biochemistry in biosafety;

• Microbiology, Parasitology and Virology in Biosafety;

• DNA barcoding and species determination;

• Real-Time PCR and quantitative determination of hidden species in mixtures and / or processed foods;

• Miniaturization of tools and field analysis;

• Advantages and dangers of Open Science. Biohacking;

• Migration of organisms and dangers to human, animal and environmental health;

• The safety in research laboratories and the safety of research products.

AT THE END THE STUDENT WILL BE ABLE TO:

• Know the methods of scientific analysis for the management of commercial and food products;

• Know the advantages and disadvantages of open scientific information management systems;

• Have the basic knowledge to be able to face biosecurity problems, with the appropriate techniques and with experimental verification and laboratory exercises;

The course aims to broaden the knowledge on the most current biosecurity monitoring systems increasingly used in research, but also in the control of the circulation of goods, products marketed online, sophistication and fraud.

In particular, topics such as:

• Biosecurity: General and introductory concepts;

• DNA profiling and its uses;

• Chemistry and biochemistry in biosafety;

• Microbiology, Parasitology and Virology in Biosafety;

• DNA barcoding and species determination;

• Real-Time PCR and quantitative determination of hidden species in mixtures and / or processed foods;

• Miniaturization of tools and field analysis;

• Advantages and dangers of Open Science. Biohacking;

• Migration of organisms and dangers to human, animal and environmental health;

• The safety in research laboratories and the safety of research products.

AT THE END THE STUDENT WILL BE ABLE TO:

• Know the methods of scientific analysis for the management of commercial and food products;

• Know the advantages and disadvantages of open scientific information management systems;

• Have the basic knowledge to be able to face biosecurity problems, with the appropriate techniques and with experimental verification and laboratory exercises;

PREREQUISITES

Students should have basic notions of chemistry, biochemistry, cell biology and cell culture.

TEACHING METHODS

The course consists of 12 lectures of 2 hours each and 4 afternoons of 4 hours each in the laboratory

 

 

SYLLABUS/CONTENT

Lesson 1: General and introductory concepts

 

- Introduction to biosecurity. General concepts.

- Globalization, movement of goods and living organisms, Open Science and control of the use of scientific information. “DIY” genetics and biohackering, hints and commercializations.

- Integrated approach for biosafety: Why Genetics, Biology, Chemistry, Microbiology work together. Some practical cases of problems treated with integrated approaches and the need for information integration.

- Brief general introduction to the technologies and methodologies used and their uses.

- The field laboratory. Evolution of systems and tools to be transferred to the field with advantages for response time, usability and costs. Smartphone and miniaturization.

 

 

Lesson 2: DNA profiling and its uses

 

- Sampling techniques in different environments and danger of contamination.

- Different techniques of extraction of nucleic acids and proteins of animal, vegetable, bacterial or viral origin; extraction of trace molecules.

- DNA Fingerprinting. Minisatellites, Microsatellites, Rapd, various commercial STR amplification systems in eukaryotic organisms.

- Discussion of cases and possible uses in different fields: forensics, food, law and economics.

- Individual plant DNA fingerprinting. Case discussion.

 

 

Lesson 3: Chemistry and biochemistry in biosafety

 

- Chemistry and biochemistry for the identification of trace molecules.

- Techniques in use: Mass Spectrometry, Chemical Fingerprinting.

- Uses.

- Practical cases of specific proteins of some plants or animals and their identification for food safety, forensic and / or commercial purposes. Case discussion (Italian olive oil and RAMAN / NIR spectrometry). The Blend Problem.

 

 

Lesson 4: Microbiology, Parasitology and Virology at work together

 

- Microbiology, parasitology and virology in the food chain. Zoonoses. Methods used for monitoring. The prions. Discussion of cases of contamination. Discussion of cases of infection. Monitoring of epidemics in food-related species. West Nile, Sars-Cov-2, Ebola etc. African swine fever. Hypothesis of immune lowering in bats.

 

 

Lesson 5: DNA barcoding and the determination of species in biosafety

 

- Food fraud and food adulteration: Substitution of species, dilution with different species.

            Economic aspects, nutritional aspects and possible health implications. Patent infringement example.

- Species identification techniques: DNA Barcoding. Application Utilities and Case Discussion. Identification of different cultivars in the vegetable field. A case of forensic interest: the different hemp cultivars with different alkaloid contents. Adulteration of cheeses. Other examples.

 

 

Lesson 6: Real-Time PCR and quantitative determination of hidden species in mixtures and / or processed foods

 

- Identification of different species by species-specific Real-Time PCR.

            Possibility of quantitative evaluation of illegal species hidden in mixtures. The case of the Peganum Harmala.

- Use of allele-specific probes.

- Adaptogenic plants, psychotropic plants and the dissemination and marketing on the net. Ethnic drugs and NPS.

- Poisonous plants and human health problems. Cases. Track monitoring.

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Lesson 7: Miniaturization of instruments. Field analysis

 

- Mini Real-Time PCR,

- Mini-sequences

- Biosensors

- Colorimetric PCR (Nanotechnologies in the field)

- Colorimetric Narcotest

- Smartphone and lenses. The Algawarning project.

- Remote analysis

- Smartphone and slide analysis in inaccessible areas (identification of protists)

 

 

Lesson 8: from hacking to biological piracy. Dangers and remedies

 

- Open Science / Open Access: advantages and dangers.

- The crispr-cas9 system

- Biohackering. Striking cases.

- Ethnospecific possibilities

- Antibiotic resistances and kits for sale.

- Legal aspects of transgenesis

- Bioterrorism: The main biological agents.

 

 

Lesson 9: The assessment of toxicity and the identification of new substances / products that are hazardous to health

 

- In vitro toxicity assessment methods.

- MTT, Neutral reed Uptake, MEA, Narcotests, specific colorimetric assays.

- In vitro toxicology on hazardous substances.

Lesson 10: Migration of organisms and dangers to human, animal and environmental health

 

- Sequencing in biosafety.

- Nanopore sequencing and field analysis

- Identification of profiles of bacterial populations contaminated by allochthonous strains.

- Examples of imbalances and effects on human and animal health.

- Danger to native species and ecological balances with the introduction of alien eukaryotic species. LIFE Asap project.

- Example of bats and virus resistance. Discussion of animal transmission cases. (West Nile, Ebola, and others).

- Ecozone: problems for both vegetable and animal organisms of food interest. the one health approach.

 

 

Lesson 11: Safety in research laboratories and safety of research products

 

- Biosecurity of laboratory products: cells, monoclonal antibodies, RNA vaccines, liposomes. Safety of blood and its derivatives (platelet gel). Future possibilities for organoids and tissue engineering.

- Discussion of new cases presented by the students and their ideas of possible solutions.

 

 

Lesson 12: Brainstorming; at work all together

 

- Discussion of real cases presented and chosen by the students

 

EXERCISES

 

Exercise 1; (4 h): Determination of the transgenic charge of a food product

 

- It is necessary to identify and quantify a transgene in a commercial product of plant origin. The case of transgenic soy. Brief introduction to the legal and economic aspects of the case.

  

            In practice: a) Extraction of the DNA sample. B) Amplification reaction by Quantitative Real-Time PCR. 3) Analysis of the results and evaluation of the amount of transgene present in the sample.

 

Exercise 2; (4h): DNA Profiling. Engineered case fingerprinting.

 

            We will perform a DNA profile of different samples identifying the "Match" and solving the question.

 

            In practice: we will perform STR PCR and electropherogram and we will analyze the obtained profiles

 

Exercise 3; (4h): Quantitative identification of a smart drug hidden in a blend of legal herbs or in a food supplement

 

            We will analyze a blend of herbs supplement to identify and quantify the content of a dangerous species

 

            In practice: we will perform DNA extraction and. a series of Real-Time PCR by building a calibration of the system and verifying the percentage of the species hidden in the mixture

 

Exercise 4; (4h): Quantitative identification of a species in a processed product and verification of what is declared on the label.

 

            We will analyze processed and processed food products to highlight any species substitutions and / or species dilutions that constitute commercial fraud and potential health hazards by performing species-specific PCR reactions.

            In practice: we will use species-specific PCR reactions to highlight adulterations in commercial samples.

RECOMMENDED READING/BIBLIOGRAPHY

 

The material will be provided by the teacher and uploaded on aulaweb