UNIVERSITY OF VETERINARY AND PHARMACEUTICAL SCIENCES BRNO FACULTY OF VETERINARY MEDICINE Department of Pharmacology and Pharmacy PRACTICAL LESSONS FROM VETERINARY PHARMACOLOGY SMALL ANIMALS EXPERIMENTS MVDr. Simona Pejřilová, PhD. Department of Pharmacology and Pharmacy BRNO 2011
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UNIVERSITY OF VETERINARY AND PHARMACEUTICAL SCIENCES BRNO
FACULTY OF VETERINARY MEDICINE
Department of Pharmacology and Pharmacy
PRACTICAL LESSONS FROMVETERINARY PHARMACOLOGY
SMALL ANIMALS EXPERIMENTS
MVDr. Simona Pejřilová, PhD.Department of Pharmacology and Pharmacy
BRNO 2011
Preface
Welcome to the first edition of Practical Lessons from Veterinary Pharmacology, Small
Animals Experiments. It´s dedicated to the veterinary students learning veterinary
pharmacology as a part of their quest to become doctors of veterinary medicine.
Main goal of this textbook is to contribute to the right way of the experiments realiza-
tion, correct and humanly approach to experimental animals.
Simona Pejřilová
Brno, 2011
Obsah
Introduction to the Textbook ............................................................................................... 5Experimental Animals and their Physiological Parameters ................................................ 6Drug Doses ........................................................................................................................... 8Summary of Experiments ................................................................................................... 9EXPERIMENT No. 1 all groups ......................................................................................... 10EXPERIMENT No. 2 version A .......................................................................................... 13EXPERIMENT No. 2 version B .......................................................................................... 15EXPERIMENT No. 3 version A .......................................................................................... 17EXPERIMENT No. 3 version B .......................................................................................... 20EXPERIMENT No. 4 version A .......................................................................................... 23EXPERIMENT No. 4 version B .......................................................................................... 27EXPERIMENT No. 5 version A .......................................................................................... 31EXPERIMENT No. 5 version B .......................................................................................... 34EXPERIMENT No. 6 version A .......................................................................................... 37EXPERIMENT No. 6 version B .......................................................................................... 40EXPERIMENT No. 7 all groups ......................................................................................... 43EXPERIMENT No. 8 all groups ......................................................................................... 45EXPERIMENT No. 9 all groups ......................................................................................... 47EXPERIMENT No. 10 all groups ....................................................................................... 50EXPERIMENT No. 11 all groups ....................................................................................... 54EXPERIMENT No. 12 all groups ....................................................................................... 57References ......................................................................................................................... 59Abbreviations ..................................................................................................................... 60
Introduction to the Textbook
How to use the Textbook
This textbook is intended for the students of 3rd Study Year of the Veterinary Univer-
sity, topic Veterinary Pharmacology.
Textbook detail specifies 12 experiments, which are carrying out on small laboratory
animals, namely laboratory rats and domestic rabbits.
Experiments are focused on these general topics: General and Local Anaesthesia,
Opioid Analgesics, Analeptics, Drugs with effects on Cardiovascular System and
Blood, Drugs affected Sympathetic and Parasympathetic system. Besides that, there
is one experiment (the first one), which is pointed at basic steps describing the ma-
nipulation with above mentioned animals.
Each experiment is divided into basic parts:
• Examination of the experimental animals in detail
• Task
• Necessary
• Procedure
• Results
• Conclusion
• Prescriptions
Basic Rules for Practical lessons:
• Read the manual first and study specified Topic!
• Keep the “three Rs” as guiding principles for manipulation with Experi-mental Animals in Research! (Refinement, Reduction, Replacement)
5
Experimental Animals and their Physiological Parameters
Laboratory Rat. Rattus norvegicus.
Order: Rodentia.
Family: Muridae.
General Physiological Parameters
Body Temperature°C
Respiratory RateBreaths/min
Heart RateBeats/min
37.5—39.0 70—110 250—500
Weight of Adult 220—500 g Length of Gestation 21—26 days
Length of Life 2—3 years Weight of Newborn 5—7 g
Sexual Maternity 5—8 weeks Length of Breast-feeding 21 days
6
Domestic Rabbit. Oryctolagus cuniculus.
Order: Lagomorpha.
Family: Leporidae.
General Physiological Parameters
Body Temperature°C
Respiratory RateBreaths/min
Heart RateBeats/min
38.5—40.0 30—60 180—325
Weight of Adult 2—6 kg Length of Gestation 29—33 days
Length of Life 3—7 years Weight of Newborn 30—80 g
Sexual Maternity 4—7 months Length of Breast-feeding 4—7 weeks
7
Drug Doses
We focus on drug doses of some chosen active substances which are used in our
practical lessons in this part of textbook.
Adrenaline (Epinephrine)
Rabbits: 6—9 mg/kg IV or 9 mg/kg IM, other rodents: 20 mg/kg IM or
5 % eye drops with cocaine. Prescribe total amount 10 ml, for your practice.
Cocaine hydrochloride belongs to Dependency Producing Substances (Narcotic
Substances, Group 1).
16
EXPERIMENT No. 3 version A
Date:
Sympathomimetic effect of Adrenaline on the Blood Pressure
of Rabbit
Species:
Personal data:
TRIAS:
Body Temperature°C
Respiratory RateBreaths/min
Heart RateBeats/min
Physical Examination:
Task:
Measure the blood pressure changes of the rabbit by non-invasive method over the
time, observe the shape of the curve and register the changes after administration of
Adrenaline.
Necessary:
• 1 rabbit;
• Grant´s measuring device;
• syringes; needles;
• MPMP: Adrenalin Léčiva ®.
Procedure:
Measure the change of the blood pressure on Arteria auricularis centralis of rabbit.
17
Measure it five times and calculate average value for elimination of mistake.
Administer Adrenaline 0.09 ml/kg IM.
Calculate the dose: Adrenaline 0.09 ml/kg. CHECK dose with assistant!
Measure the blood pressure at these intervals: 1 min., 3 min., 5 min., 10 min.,
15 min., 30 min. and 45 min. after administration of the active substance.
Always measure five times and calculate the average value!
Tab. No. 1.
Blood pressure (mm Hg)
before 1´ 3´ 5´ 10´ 15´ 30´ 45´
Plot results into the graph:
Graph No. 1.
x, Blood pressure (mm Hg)
y, Time (min)
Other changes:
18
Conclusion:
Prescriptions:
MPMPs:
Adrenalin Léčiva ®, solution for injection. Prescribe 5 original packages, for
your practice.
19
EXPERIMENT No. 3 version B
Date:
Sympathomimetic effect of Ephedrine on the Blood Pressure
of Rabbit
Species:
Personal data:
TRIAS:
Body Temperature°C
Respiratory RateBreaths/min
Heart RateBeats/min
Physical Examination:
Task:
Measure the blood pressure changes of the rabbit by non-invasive method over the
time, observe the shape of the curve and register the changes after administration of
Ephedrine.
Necessary:
• 1 rabbit;
• Grant´s measuring device; syringes; needles;
• MPMP: Ephedrin Biotika ®.
20
Procedure:
Measure the change of the blood pressure on Arteria auricularis centralis of rabbit.
Measure it five times and calculate average value for elimination of mistake.
Administer Ephedrine 5 mg/kg IM.
Calculate the dose: Ephedrine 5 mg/kg. CHECK dose with assistant!
Measure the blood pressure at these intervals: 1 min., 3 min., 5 min., 10 min.,
15 min., 30 min. and 45 min. after administration of the active substance.
Always measure five times and calculate the average value!
Tab. No. 1.
Blood pressure (mm Hg)
before 1´ 3´ 5´ 10´ 15´ 30´ 45´
Plot results into the graph:
Graph No. 1.
x, Blood pressure (mm Hg)
y, Time (min)
Other changes:
Conclusion:
21
Prescriptions:
MPMP:
Ephedrin Biotika ®, solution for injection. Prescribe 3 original packages, for
your practice.
22
EXPERIMENT No. 4 version A
Date:
Mydriasis and Miosis
Species:
Personal data:
TRIAS:
Body Temperature°C
Respiratory RateBreaths/min
Heart RateBeats/min
Physical Examination:
Task:
Recognize the effects of Physostigmine and Atropine on the diameter of the pupil of a
rabbit.
Necessary:
• 1 rabbit;
• 0.05 % eye drops with sulphate of atropine;
• 1 % eye drops with salicylate of physostigmine.
Procedure:
1st part:
Measure size of pupil before experiment by paper gauge.
Administer 4 drops of eye drops with Physostigmine into right conjunctival sac.
Left eye of rabbit is for control.
23
Observe changes on eyes (size of pupils) and other changes on rabbits for 45 min.
Tab. No. 1.
TimePhysostigmine
left eye right eyebefore administrativ3 min after administration5 min after administration15 min after administrativ30 min after administrativ45 min after administrativ
Other changes on the eyes:
Other changes (GIT, RT) – GIT:
Other changes – RT:
2nd part:
Make opposite procedure after 45minutes:
24
Administer 4 drops of eye drops with Atropine into right conjuctival sac (the same
treated eye).
Observe changes of pupils and other parts of organism after re-administrations for
20 minutes.
Tab. No. 2.
TimeAtropine
left eye right eye
Administration 2nd drug
3 min after administration
5 min after administration
15 min after administrativ
20 min after administrativ
Please, rinse out rabbit´s eye by Ophtal (MPMP) after finishing of experi-
ment!
Other changes on the eyes:
Other changes (GIT, RT) – GIT:
Other changes – RT:
25
Conclusion:
Prescriptions:
IPMPs:
0.05 % eye drops with sulphate of atropine, 10 ml. For patient.
1 % eye drops with salicylate of physostigmine, 20 ml. For your private
practice.
26
EXPERIMENT No. 4 version B
Date:
Mydriasis and Miosis
Species:
Personal data:
TRIAS:
Body Temperature°C
Respiratory RateBreaths/min
Heart RateBeats/min
Physical Examination:
Task:
Recognize the effects of Physostigmine and Atropine on the diameter of the pupil of a
rabbit.
Necessary:
• 1 rabbit;
• 0.5 % eye drops with sulphate of atropine;
• 1 % eye drops with salicylate of physostigmine.
Procedure:
1st part:
Measure size of pupil before experiment by paper gauge.
Administer 4 drops of eye drops with Atropine into right conjuctival sac.
Left eye of rabbit is for control.
27
Observe changes on eyes (size of pupils) and other changes on rabbits for 45 min.
Tab. No. 1.
TimeAtropine
left eye right eye
before administration
3 min after administration
5 min after administration
15 min after administration
30 min after administration
45 min after administration
Other changes on the eyes:
Other changes (GIT, RT) – GIT:
Other changes – RT:
2nd part:
Make opposite procedure after 45minutes:
Administer 4 drops of eye drops with Physostigmine into right conjunctival sac (the
same treated eye).
28
Observe changes of pupils and other parts of organism after re-administrations for
20 minutes.
Tab. No. 2.
TimePhysostigmine
left eye right eye
Administration 2nd drug
3 min after administration
5 min after administration
15 min after administration
20 min after administration
Please, rinse out rabbit´s eye by Ophtal (MPMP) after finishing of experi-
ment!
Other changes on the eyes:
Other changes (GIT, RT) – GIT:
Other changes – RT:
29
Conclusion:
Prescriptions:
IPMPs:
0,5 % eye drops with sulphate of atropine, 10 ml. For patient.
1 % eye drops with salicylate of physostigmine, 20 ml. For your private
practice.
30
EXPERIMENT No. 5 version A
Date:
Antagonistic Effect of Atropine and Pilocarpine on Salivary
Glands and Pupil Size
Species:
Personal data:
TRIAS:
Body Temperature°C
Respiratory RateBreaths/min
Heart RateBeats/min
Physical Examination:
Task:
Recognize the effects of Pilocarpine and Atropine on the rabbit´s Salivary Gland.
Necessary:
• 1 rabbit;
• glass cylinder; funnel;
• IMPM: 1 % eye drops with sulphate of atropine; 1 % eye drops with hydro-
chloride of pilocarpine.
Procedure:
Administer 5 drops of eye drops with Piocarpine into left conjuctival sac and adminis-
ter 5 drops of eye drops with Atropine into right conjuctival sac.
Collect and measure amount of saliva in glass cylinder in ml every five minutes.
31
Tab. No. 1.
SALIVA (ml) 5 min 10 min 15 min 20 min 25 min 30 min
Atropine
Pilocarpine
Tab. No. 2.
Pupil size in mm beforeadministration
afteradministration
Left eye-Pilocarpine
Right eye- Atropine
Please, rinse out rabbit´s eye by Ophtal (MPMP) after finishing of experi-
ment!
Other changes on the eyes:
Other changes (GIT, RT) – GIT:
Other changes – RT:
32
Conclusion:
Prescriptions:
1 % eye drops with sulphate of atropine, 10 ml.
1 % eye drops with hydrochloride of pilocarpine, 10 ml.
For your private practice.
Prescribe these 2 MPs one 1 form for prescription.
33
EXPERIMENT No. 5 version B
Date:
Effect of Pilocarpine on Salivary Glands and Pupil Size
Species:
Personal data:
TRIAS:
Body Temperature°C
Respiratory RateBreaths/min
Heart RateBeats/min
Physical Examination:
Task:
Recognize the effects of Pilocarpine on the rabbit´s Salivary Gland.
Necessary:
• 1 rabbit;
• glass cylinder; funnel;
• IMPM: 1 % eye drops with hydrochloride of pilocarpine.
Procedure:
Administer 5 drops of eye drops with Pilocarpine into left and right conjuctival sac
(into both conjuctival sacs).
Collect and measure amount of salivas in glass cylinder in ml every five minutes.
34
Tab. No. 1.
SALIVA (ml) 5 min 10 min 15 min 20 min 25 min 30 min
Pilocarpine
Tab. No. 2.
Pupil size in mm beforeadministration
afteradministration
Left eye
Right eye
Please, rinse out rabbit´s eye by Ophtal (MPMP) after finishing of experi-
ment!
Other changes on the eyes:
Other changes (GIT, RT) – GIT:
Other changes – RT:
Conclusion:
35
Prescriptions:
1 % eye drops with sulphate of atropine, 10 ml.
1 % eye drops with hydrochloride of pilocarpine, 10 ml.
For your private practice.
Prescribe these 2 MPs one 1 form for prescription.
36
EXPERIMENT No. 6 version A
Date:
Opposite effects of Parasympathomimetic- Carbachol and Para-
sympatholytic- Atropine on the Function of Digestive Tract
Species:
Personal data:
TRIAS:
Body Temperature°C
Respiratory RateBreaths/min
Heart RateBeats/min
Physical Examination:
Task:
Recognize the stimulating effect of Carbachol on peristalsis and saliva secretion and