The small introduction into practial training

Department of pathophysiologyBuilding 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 experimentLaparotomy 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

research(15% )

toxicitytesting(15 % )

vaccinetesting(15 % )

pharmaceuticalindustry(25 % )

others[diagnostics,

exp. medicine,education]

(30 % )

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 20th

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 20th century– experimental animals on its

own specialised breeding of

laboratory animals inbreed and outbreed strains development of new strains alternative exp. models

– from 80th of the 20th 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

bezmikrobníA

A+B

A+B+C

amicrobic(axenic)

monoxenic

polyxenic

dixenic

conventional

without specifiedpathogens

-P1-P2-P3

DEFIN

ED

AS

SO

CIA

TIO

N

GN

OTO

BIO

TIC

SPF

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)