Blood sample collection in lab. animals

Handling of laboratory animals
&
Blood sample collection in small laboratory
animals
Subramani Parasuraman, M.Pharm, Ph.D.,
Associate Professor,
Unit Head, Unit of Pharmacology,
Faculty of Pharmacy,
AIMST University, Malaysia
Email: parasuraman@aimst.edu.my

Handling of laboratory animals (mice)
• Mice and rats are both highly adaptable animals and
generally easy to restrain.
07-09-2018
Symposium on“The fundamentals of animal
research” JIPMER, PDY
2

Handling of laboratory animals (Rats)
• Rats are basically docile, gentle handling and curious
animals that respond to quiet. They are not usually
aggressive.
07-09-2018
3

Handling of laboratory animals (Guinea pig)
• The handler should be rapid and smooth, to avoid
frightening the animal.
07-09-2018
4

Handling of laboratory animals (Rabbit)
• Rabbits are especially susceptible to the effects of
stress and should always be approached in a calm
and confident manner.
07-09-2018
5
The rabbit should be
held its head tucked
under the handler’s
arm and with the back
and hindquarters
supported by the
handler’s forearms.

Blood sample collections
07-09-2018
6

General Principles
• The method of blood collection should be described in
the protocol approved by the Institute animal ethics
committee.
• In general, blood sample is withdrawn from venous,
arterial blood vessels or heart chambers.
• It should be least painful and stressful.
• Adequate training is required for blood collection using
any method in any species.
• Frequency of blood collection is important. Once in two
weeks is ideal for rodents. If the study needs multiple
blood samples, lagomorphs (e.g., hares and rabbit) can
be used.
07-09-2018
7

General Principles
• All nonterminal blood collection without
replacement of fluids is limited up to 10% of total
circulating blood volume in healthy, normal, adult
animals on a single occasion and collection may be
repeated after 3 to 4 weeks.
• The estimated blood volume in adult animals is 55
to 70 ml/kg body weight. If blood collection volume
exceeds more than 10% of total blood volume, fluid
replacement may be required.
07-09-2018
8

General Principles
• The amount of blood in the human body is generally
equivalent to 7 – 8 % body weight
• Weighing 45 kg (3.2 – 3.6 L) of adult can safely donate one
unit (350 mL) of blood.
Approximately 10% of total blood volume
Approx. BW: 20 – 30 g
Blood volume: 1.4 – 2 mL
Max. vol. of sample: 0.15 – 0.2 mL
Approx. BW: 0.7 – 1.2 kg
Blood volume: 50 – 90 mL
Max. vol. of sample: 5 – 9 mL
Approx. BW: 200 -250 g
Max. vol. of sample: 1 – 1.5 mL
Approx. BW: about 1 kg
Max. vol. of sample: 7 – 8 mL
07-09-2018
9

Blood sample collection in rats and mice
07-09-2018
10

Blood samples are collected using the following
techniques
• Blood collection not requiring anesthesia
• Saphenous vein (rat, mice, guinea pig)
• Dorsal pedal vein (rat, mice)
• Blood collection requiring anesthesia (local/general anesthesia)
• Tail vein (rat, mice)
• Tail snip (mice)
• Orbital sinus (rat, mice)
• Jugular vein (rat, mice)
• Temporary cannula (rat, mice)
• Blood vessel cannulation (rat, guinea pig, ferret)
• Tarsal vein (guinea pig)
• Marginal ear vein/artery (rabbit)
• Terminal procedure
• Cardiac puncture (rat, mice, guinea pig, rabbit, ferret)
• Orbital sinus (rat, mice)
• Posterior vena cava (rat, mice)
07-09-2018
11

Rat blood circulation
07-09-2018
12

General methods for blood collection
• Blood collection not requiring anesthesia
– Saphenous vein
– Dorsal pedal vein
07-09-2018
13

General Methods for Blood sample Collection
• Dorsal pedal vein
07-09-2018
14

• Blood collection requiring anesthesia
– Tail vein
– Orbital sinus
– Jugular vein
– Temporary cannula
– Blood vessel cannulation
07-09-2018
15

• Tail vein
07-09-2018
16

• Tail vein
Thrombosis
07-09-2018
17

• Terminal procedure
– Cardiac puncture
– Orbital sinus
– Posterior vena cava
07-09-2018
18

Orbital sinus
Rat eye
Human eye
• Rats have poor visual acuity (20
times worse than humans)
• Rats have rods and two types of
cones which detects GREEN and
BLUE colour.
• Rats are unable to change their
lenses’ shape because they have
poorly developed ciliary muscles.
Parasuraman S, Zhen KMing, Raveendran R. Retro-orbital sample collection in Rats - a video
article. PTB Rep. 2015;1(2):37-40.
07-09-2018
19

Orbital sinus
07-09-2018
20

Orbital sinus
07-09-2018
21

Orbital sinus
Video: Blood sample collection from rat orbital sinus using left predominant hand
Blood sample collection from rat orbital sinus using right predominant hand
07-09-2018
22

Orbital sinus - ADR
07-09-2018
23

Adverse effects of blood sample collections
• Stress, haemorrhage, bruising, thrombosis,
infection at the site of needle entry, phlebitis,
scarring, and nerve damage are some of the
potential adverse effects which should be avoided.
• Venipuncture may cause ‘bruising’, so proper
monitoring is required.
• Failure in aseptic procedures may cause
thrombosis and phlebitis.
07-09-2018
24

Blood sample collection in rabbits
• Marginal ear vein/artery
• Cardiac puncture
07-09-2018
25

Blood sample collection in rabbits
• Marginal ear vein/artery blood sample collection
Sample collection form marginal ear vein
Sample collection form artery
07-09-2018
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Blood sample collection in guinea pigs
• Saphenous vein
• Tarsal vein
• Blood vessel
cannulation
• Cardiac puncture
07-09-2018
27

Blood sample collection in guinea pigs
• Sample collection from tarsal vein
07-09-2018
28

Blood groups in Rats
Ref: Burhoe SO. Blood Groups of the Rat (Rattus Norvegicus) and Their Inheritance. Proc Natl Acad Sci U S A. 1947
May;33(5):102-9.
Blood groups in Rabbits
Ref: Carl Cohen. Blood groups in rabbits. annals of
the new york academy of sciences. 1962
07-09-2018
29

for more reading………..
• Parasuraman S, Raveendran R, Kesavan R. Blood
sample collection in small laboratory animals. J
Pharmacol Pharmacother 2010;1:87-93.
• Parasuraman S, Zhen KM, Raveendran R. Retro-
orbital sample collection in rats – A video article.
Pharmacol Toxicol Biomed Rep. 2015;1:37–40.
07-09-2018
30

Thank you
with his guidance..
• Obtain my Ph.D.,
• Published paper which got more
number citations
• Become a Editor-in-Chief JYP,
JPNR
• Transferred knowledge to more
than 600 undergraduate students
and practical skill to 60
undergraduate research
students, 20 postgraduate
students, 5 doctorate students.
07-09-2018
31

Blood sample collection in lab. animals

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