66633 - Biotechnology and Animal Models

Learning outcomes

To prepare students for experimental work with live animals.

The course will focus on the use of animals in biomedical research. Based uponFELASA Programit should provide an introduction to the major areas of laboratory animal science and in this way contribute both to the humane use and care of animals and to the quality of experimental results.

The student must have knowledge of male and female gametogenesis in order to know and understand principal biotechnologies, comprising transgenesis, applied to male and female gametes and embryos of domestic, terrestrial and aquatic, species.

Course contents

The course is composed by a theory part and a pratical laboratory work.

The first theory part is devoted to Laboratory Animal Science as backgroud to perform experiments involving animals (Prof Forni)

1.     Ethical and historical aspects.

2.     Alternatives (3R's : Replacement, reduction, Refinement)

3.     Husbandry, nutrition and care

4.     Biology and physiology

5.     Behaviour, stress and well-being

6.     Genetical and Microbiological standardization

7.     Quality management, good laboratory practices and standard operating procedures

8.     Animal Models for Biomedical Research

The second theory part is devoted to physiology of reproduction and reproductive biotechnologies applied to domestic, terrestrial and aquatic, species.

1.   Outline of reproductive function regulation. Reproductive hormones (hypothalamus-hypophysis-gonad axis) and endocrinological function of gonads

2.   Female. Ovarian function: oogenesis, folliculogenesis, ovulation and corpus luteum.

3.   Male. Testicle function: spermatogenesis. Male gametes: characteristics and function.

4.   Embryos:  Fertilization. First phases of embryo development till blastocyst stage.

5.   Female gamete: in vivo and ex vivo oocyte collection; oocyte qualitative evaluation; techniques for in vitro growing oocyte; oocyte in vitro maturation, micromanipulation and cryoconservation.

6.   Male gamete: in vivo and ex vixo (epididimal) spermatozoa collection; quali-quantitative semen analysis, dilution and cryoconservation of spermatozoa by refrigeration/freezing/vitrification. Functional characteristic evaluation of  spermatozoa. Fertilization techniques with frozen or fresh semen, artificial insemination, laparotomic/laparoscopic intrauterine insemination. Spermatozoa sexing.

7.   Embryos: surgical and non surgical in vivo collection, in vitro production by in vitro fertilization; in vitro colture; embryotransfer; micromanipulation: splitting, nuclear transfer; cloning; stem cell; sexing. Transgnenesis.

The practical work will utilize  swine or bovine as species of reference.

1.   Female Gamete: ex vivo oocyte collection; qualitative oocyte evaluation before in vitro maturation; oocyte in vitro maturation; evaluation of oocytes after fixation.

2.   Male gamete: in vivo and ex vixo (epididimal) spermatozoa collection; quali-quantitative semen analysis, dilution and cryoconservation of spermatozoa by refrigeration. Functional characteristic evaluation of  spermatozoa.

3.   Animal transgenesis by Sperm Mediated Gene Transfer:  diluent preparation, semen manipulation, DNA uptake procedures, Evaluation of DNA uptake by PCR.

As concerns the teaching methods of this course unit, all students must attend Module 1, 2 online, while Module 3 on health and safety is to be attended in class. Information about Module 3 attendance schedule is available on the website of your degree programme

 

Readings/Bibliography

Lesson notices;
L. F. van Zutphen, V. Baumans, A. C. Beynen. Principles of Laboratory Animal Science, Revised Edition Edited By, Utrecht University, Department of Veterinary Science, Utrecht, The Netherlands
Reproduction in Farm Animals 7th edition, E. S. E. Hafez, B. Hafez (Editors) 2000;
Lippincott Wlliams and Wilkins Dukes, Fisiologia degli animali domestici, IDELSON-GNOCCHI, Napoli (2002).
Lab handouts with the operational descirption of the experiences

Teaching methods

Lectures and cooperative learning

Laboratory work performed personally by the student according to the handouts provided, with theoretical and practical explanations of the teacher and tutor

Assessment methods

The principal learning outcomes will be assessed:

- knowledge on the use of animals in biomedical research.

- judgement autonomy and communication tools in relation to animal experiments

- knowledge of male and female gametogenesis

- knowledge on reproductive biotechnologies

The final grade will be composed of:
40% on a multiple choice quiz (32 questions) on Laboratory Animal Science at the end of the first part of the course;
10% on the active participation during lecture and laboratory activities;
5% on the report on laboratory activity (to be send by mail at the exam list registration);
45% on oral discussion about lecture topics.

Teaching tools

Lectures .ppt

Laboratories of bioinformatic, cell coltures, microbiology and morphology

Office hours

See the website of Monica Forni

See the website of Maria Laura Bacci