The small introduction into practial training Department of pathophysiology Building A18

Must have  White lab coat  Overshoe or slipper  Hairband

Must know  One absence per semester is permited (without written excuse)  Group swap – permitted by teacher only  Experimental records (protocols)  Health and safety in the lab –Protective aids –If you are unsure - then ask your teacher

Principals of use of laboratory animals, ethics, scientific experiment Laparotomy and inspection of abdominal cavity

Common species of lab. animals  Invertebrates –Saccharomyces cerevisiae –Drosophila melanogaster –Caenorhabditis elegans  Vertebrates –Danio rerio –mouse (Mus musculus var.) –rat (Rattus norvegicus var.)85% –guinea pig (Cavia porcellus) –hamster (Mesocricetus auratus) –rabbit (Oryctolagus cunniculus)~1% –dog (Canis familiaris) –cat (Felis catus)<1% –quail (Coturnix coturnix) –primates (Macacus rhesus)<0.5%

Experimental organisms

Exp. animals used per year  globally –~50 mil./year  Europe –~12 mil./year

Controversy regarding lab. animals  for –virtually every medical achievement in 20th and 21st century involved animals in some way  each of us is a consumer of some of the many outcomes which came from animal research –that humans have obligations to ourselves that they do not have to animals  against –animal research is cruel and unjustifiable even when providing benefit for humans since they can be seen as yet another of many species inhabiting this world with no superior moral rights  we could probably benefit the same way from experiments performed by Nazis on humans –all living organisms have the same rights in the nature

History of the use of animals in medicine  until first half of 20 th century –dead animals (mainly domestic or stray animals = dissection)  Greeks (Corpus Hippocraticum -400 př. n. l.)  Galen of Pergamon  renaissance –live animals (anaesthesia = functions in vivo)  W. Harvey, M. Malpighi, Ch. Darwin, R. Koch, L. Pasteur, F. Banting & G. Best,...  C. Bernard (1867)  “Introduction to the experimental medicine”  after second half of the 20 th century –experimental animals on its own  specialised breeding of laboratory animals  inbreed and outbreed strains  development of new strains  alternative exp. models –from 80 th of the 20 th cent.  transgenic

Research use of animals – role of experiment  Evidence-based medicine –knowledge/learning  empiricism  science  Observation and experiment are tools of understanding –observation  looking at nature as we find it  the more sophisticated understanding the more variables can be looked concurrently  we can measure but we can’t reproduce –experiment  controlling nature and observing how it response to stimuli  the more sophisticated understanding the more variables can be controlled concurrently  can be reproduced

Use of animals in education  we don’t perform experiments to prove already proven but to: –understand principles of experimental work –be able to critically evaluate experimental results of others –event. be able to conduct experiments ourselves in the future

Ethics – “3R” concept  Replacement –use of alternative methods whenever possible  Reduction –minimal number necessary  Refinement –treat animals as “humanly” as possible

Alternative methods

Genetics of lab. animals  outbred –every specimen is genetically unique  inbred (=syngenic) –breeding between related subjects (over 20-x and more between given sibling pair) in order to increase homogeneity  congenic –different in solely 1 locus (mutation)  recombinant –cross-breeding between 2 inbred strains  genetically manipulated –transgenic –knock-out

Microbiologic classification

Regulation of use of lab. animals  International, EU and national level of legislation –“Animal protection law” (Czech Rep.) #246/1992 Sb., 162/1993 Sb., 167/1993 Sb., 77/2004 Sb.,... –National committee for the animal protection (Ministry of agriculture) –Ethical committees in particular institutions –Continuous education for professionals (#§17 of low 246/1992 Sb.) –autoregulation!!

Practicals  Preparation of the animal for the experiment –principles of manipulation with animal  Anesthesia –introduction (ether inhalation)  general anesthesia (i.p. mixture of ketamine + xylazine)  Laparotomy –section through 2 separate layers (skin  muscle layer) –topography of the abdominal cavity  Basic principles of surgical techniques –suture of the wound in 2 layers: continuous suture (muscle layer)  separate stitches (skin)