72711 - Biotechnologies Applied to Animal Reproduction, Genetic Engineering and Cloning

Learning outcomes

The student will learn developmental biology epigenetics of gametes and embryos in physiological and pathological conditions. The student will be able to perform collection, handling, preservation of gametes and embryos of livestock species. Assisted reproductive techniques, sexing of embryos, nuclear transfer cloning. Genetic engineering of livestock species. Set up and management of laboratory and equipment for assisted reproduction. The student will learn to work in a team and work in a laboratory. He/she will be able to understand the impact on health and welfare on the animals of the techniques both for agriculture and biomedical applications.

Course contents

Prerequisites: The student who accesses this teaching already knows the aspects related to the normal morphology and physiology of the male and female reproductive system as well as the basics of embryology. It is necessary that the student has the understanding of the spoken and written English language to allow the reading of slides (which the teacher could use in class) and of extracts from publications, scientific and manual texts, as well as watching videos.

Since this is a Master's Degree Program with students from different three-year courses and from different locations, the first day of the course will be submitted to students with a self-assessment questionnaire of previous knowledge in the topics that will be addressed in the course and of interest. The topics that will already be acquired will be discussed with Kahoot during the lectures.

The course program is structured as follows:

Laboratory and equipment management necessary for assisted reproduction, quality control. National and international legislation on animal reproduction.Oestrous detection, induction of ovulation in livestock species. Collection and preservation of male gametes. Sperm evaluation. Artificial insemination in livestock animals. In vivo embryo production by superovulation and uterine flushing.Collection, classification, handling and preservation of female gametes.n vitro embryo production: in vitro oocyte maturation, in vitro fertilization, microfertilisation (ICSI), embryo culture. Morphological evaluation, grading, sexing, cryopreservation and transfer of embryos. Embryo development and epigenetic mechanisms. Embryonic and fetal loss. Maternal recognition of pregnancy. Pregnancy maintenance. Pregnancy diagnosis.  Principles and techniques of animal cloning by nuclear transfer. Genetic engineering of large animals, potential and practical applications. Genetic engineering of animal models of human diseases, for protein of pharmacological use or for organs and tissues for xenotransplantation. Effects on health and welfare of animals. Regulatory aspects.

The program in detail (Syllabus) is available on the Study Program website.

Readings/Bibliography

The teaching material consists of the recommended textbooks, and the material (Slides presented in lessons updated every year; scientific articles and reviews found by the teacher in international journals and relevant to the learning of the subject) made available to students in the a specific website https://virtuale.unibo.it/

Reading the articles provided by the teacher is strongly recommended for the purpose of preparing for the exam.

1. Hafez, B & Hafez ESE. Reproduction in farm animals, 7th Edition. Lippincott Williams & Wilkins, 2008. ISBN 0-683-30577-8

2. Gordon, I. Reproductive Technologies in Farm Animals, CABI-Publishing, CAB International, Wallingford, Oxfordshire, UK, 2004. ISBN 0 85199 862 3.

3. Spelta, R & Corbella, E. Tecniche di fecondazione artificiale. CittàStudiEdizioni / UTET Diffusione srl, 2003. ISBN 88-251-0118-X.

4. Gordon, I. Laboratory Production of Cattle Embryos, 2nd Edition. Biotechnology in Agriculture Series, No 27 Ireland , September 2003, 592 pages HB 0851996663

5. McKKinnon A.O.,Voss J.L., Equine Reproduction , Lea & Febiger, Philadelphia 1997.

Teaching methods

The teaching consists of lectures and practical lessons.
During the lectures, the theoretical issues connected with assisted reproduction in the veterinary field and the related technologies in the main breeding and pet species will be presented and discussed.
At the end of the lectures, the students divided into groups of up to 15 people will take part in a week of practical exercises in the laboratory. During this week the student will put into practice what he learned in class within the limits of the available equipment. In particular, the following methods will be performed:

• Collection of equine and bovine oocytes from ovaries of slaughtered animals by follicular aspiration and scraping
• Oocyte classification and in vitro maturation
• Decumulation, staining and evaluation of post maturation oocytes by Hoecst fluorescence staining
• Gradient live/dead sperm separation
• epididymal sperm isolation
• Embryo freezing and vitrification

In order to carry out the practice hours, the student must have already individually studied what was presented in class regarding the techniques listed above.
To access the educational laboratory it is mandatory to wear a coat and suitable footwear. The PPE suitable for carrying out the specific activities will be provided, such as disposable latex gloves.
During the hours of exercise, a didactic excursion is scheduled at the National Institute of Artificial Insemination (INFA). To access the INFA it is necessary to wear suitable footwear and clothing.
At the end of the course there will be a moment of collective review of the topics covered.

Assessment methods

The learning verification test includes the evaluation of a group work and a final exam which consists of an oral interview. Given its role of certifying the student's skills and knowledge, the assessment will take place at the end of the learning process (end of the lessons).

Students, in groups of 5/6, once the date has been agreed with the teacher, will have to present a research project they have designed on the basis of what was covered in class and read in the bibliography. The presentation of the project must be carried out with slides divided as follows: bibliography, scope, materials and methods, expected results, possible conclusions. A facsimile of the project is made available to students on the dedicated website https://virtuale.unibo.it/

 
Through the proposed work the student learns the following skills:
- carry out a bibliographic research;
- critically read a scientific article
- set up a research project
-Team work.

Group work is mandatory. When verifying learning, the teacher will take into account the work done both in drafting the questions and in the final grade (score from 0 to 5 based on group work ability, presentation, understanding of the subject.)

During the oral interview, the teacher will ask questions aimed at ascertaining the student's theoretical/practical knowledge and critical analysis of what was presented in class and carried out during the exercise hours. The teacher can start from the request for a description of an image (oocytes, spermatozoa, embryos, devices) to deepen the topic being evaluated.

Average duration of the exam: 20 minutes.

The final grade will take into account the student's demonstration of having acquired the required knowledge and skills, the ability to connect the various topics, the possession of an expressive and language mastery. The evaluation attributed to the group work as described above will be part of the final grade.
The exam is considered passed when the student obtains an evaluation greater than or equal to the sufficiency (minimum learning objectives) equal to 18/30.

In evaluating the exam and in assigning the final grade, the teacher refers to the following "learning evaluation scale" in which the descriptors for each grade level are declined in order to make the meaning of each grade awarded.

Mark: < 18 insufficient - Fragmentary and superficial knowledge of the contents, errors in applying the concepts, deficient exposition
Score: 18 - 20 - Sufficient but general knowledge of the contents, simple exposition, uncertainties in the application of theoretical concepts
Score: 21 - 23 - Appropriate but not in-depth knowledge of the contents, ability to apply the theoretical concepts, ability to present the contents in a simple way
Score: 24 - 25 - Appropriate and extensive knowledge of the contents, good ability to apply knowledge, ability to present the contents in an articulated way
Score: 26 - 27 - Accurate and complete knowledge of the contents, good ability to apply knowledge, analytical skills, clear and correct exposition
Score: 28 - 29 - Wide, complete and in-depth knowledge of the contents, good application of the contents, good capacity for analysis and synthesis, safe and correct presentation
Score: 30 - 30 cum laude - Very broad, complete and in-depth knowledge of the contents, well-established ability to apply the contents, excellent capacity for analysis, synthesis and interdisciplinary connections, mastery of exposition

The evaluation of the oral exam is communicated at the end of the interview, in order to be immediately subject to self-evaluation by the student.
The positive outcome of the oral test is recorded at the end of the oral interview, in the presence of the student, on a specific electronic (digital) report compiled with the AlmaEsami system. Once the final profit assessment test has been passed, the student acquires the university credits (ECTS) corresponding to the educational activity subject of the assessment
The negative evaluation does not involve the attribution of a grade, but only a judgment (withdrawn or rejected) reported on the electronic (digital) report compiled with the AlmaEsami system, it is not included in the curriculum, therefore it does not affect the final grade average.

Students can book themselves for the oral exam exclusively using the methods provided by the Alma Esami online system. For any information on how to book, connect to the website: (https://almaesami.unibo.it/almaesami/welcome.htm ). Exam sessions are offered within the specific windows for current students and throughout the academic year for non-current students.
Students with disabilities or specific learning disabilities e

Teaching tools

Use of PC with multimedia projector, videos, clickers, scientific publications in specialized journals, specialized web sites. For laboratory sessions specialized equipment will be used for recovery, handling, manipulation and for cell culture.

Office hours

See the website of Eleonora Iacono