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Venipuncture techniques in chelonian species

Article  in  Lab Animal · August 2008

DOI: 10.1038/laban0708-303 · Source: PubMed

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Cyndi Brown, DVM, Column Editor
Venipuncture techniques in chelonian species
Christoph Mans, med. vet.
Blood collection is one of the most common diagnostic procedures in chelonian species. Knowledge and
case-dependent selection of appropriate sampling sites are necessary to obtain a diagnostic blood sample.
Members of the order chelonian have
­s ubstantial variation in their ­v ascular
­a natomy, level of cooperation and
­potential to inflict harmful injuries. Various
­venipuncture sites have been described,
including the ­jugular vein, brachial vein,
­femoral vein, ­subcarapacial plexus, dorsal
and ventral ­coccygeal veins and occipital
sinus1,2. Sampling from the orbital sinus,
nail ­clipping and cardiocentesis have been
­reported but are not recommended. The
­clinician or ­researcher is responsible for
choosing the venipuncture site that will
cause the least amount of pain and stress and
will provide a diagnostic sample of sufficient
quality and quantity.
PREPARATION
Depending on the species, health ­status
and level of cooperation of the ­animal, it
may not be possible to access any ­suitable
­venipuncture site ­without ­sedation or ­general
­anesthesia1. The ­maximum blood volume
that can be safely ­collected from a healthy,
­normovolemic ­chelonian ranges from 0.5%
to 0.8% of the body weight (in grams)1.
A second person is usually necessary for
­assisting the primary clinician by ­restraining
the animal and ensuring ­hemostasis after
blood ­collection, by ­digital compression of the
­sampling site. The skin over the ­venipuncture
site should be ­t horoughly cleaned and
­disinfected before the ­procedure is done.
EQUIPMENT
The smallest needle size ­appropriate for
the size of the animal and ­sample volume
should be ­c hosen. In most chelonian­
species, a 22- to 27-gauge hypodermic
needle of ­suitable length is suggested. The
Tai Wai Small Animal and Exotic Hospital, Shop B-D 69-75 Chik Shun Street, Tai Wai, N.T., Hong Kong. Correspondence should be addressed to C.M. (cmans@gmx.net).
LAB ANIMAL
clinical techniques
syringe size used
will depend on the a
­v olume of blood
to b e ­c o l l e c te d .
The smallest size
sy ringe possible
should be used in
order to ­p revent
excessive negative
pressure, which can
cause ­c ollapse of b
the ­sampled vein.
PROCEDURES
Jugular vein
The jugular vein is
the blood ­sampling
site of choice in
­c helonian species
and should be used FIGURE 1 | Blood collection from the left jugular vein. (a) Anatomical site of
­whenever ­possible. the left jugular vein (arrows) in a red-eared slider (Trachemys scripta elegans).
The jugular vein T, tympanum. (b) Blood collection from the left jugular vein in an African
is visible in most spurred tortoise (Geochelone sulcata). Note the manual compression of the
neck at the level of the coelomic inlet to facilitate successful venipuncture.
­s p e c i e s a n d h a s
less risk of lymph
­c ontamination than some other sites. the ­venipuncture manually extends and
In uncooperative or ­a ggressive ­s pecies, restrains the head. Pushing the rear limbs
this sampling site is ­u nsuitable for use forward, within the shell, ­sometimes causes
­w ithout prior chemical restraint to allow the ­animal to extend its head from the shell,
sufficient extension of the head3. making it easier to grasp and restrain the
Anatomy. The (external) jugular vein head. Once the neck is extended, the ­jugular
is located superficially under the skin vein can usually be visualized under the
on both sides of the neck, running in a skin. Tightening of the cervical skin folds,
­caudodorsal direction from dorsal aspect ­a pplication of ­d isinfectant liquid and
of the ­t ympanum to the coelomic cavity. ­compression of the neck at the coelomic
In some species, a dorsal as well as a ventral inlet will ­usually enhance visibility of the
external jugular vein is present3. jugular vein. The needle is introduced in a
Procedure. The animal is positioned ­craniocaudal direction, and mild ­negative
in lateral recumbency, and the forelimbs pressure is maintained until blood is seen
are restrained. The person ­p erforming in the syringe hub (Fig. 1).
Volume 37, No. 7 | JULY 2008 303
clinical techniques
a
b
FIGURE 2 | Blood collection from the subcarapacial
sinus in a red-eared slider (Trachemys scripta
elegans). The head can either be extended and the
neck bent ventrally (a) or be pushed back into the
shell (b). A gauze square is used to prevent bite
injuries (b). Note the restraint of the forelimbs
and the positioning of the animal at a table edge,
facilitating the introduction of the needle in the
dorsocaudal direction.
Subcarapacial venous sinus
This sampling site is often used in
­u ncooperative species, when jugular
­v enipuncture is not possible without
­s edation, in neonates and other small
­chelonians, as well as in those too ill to
endure the stress of restraint or sedation.
Anatomy. The subcarapacial ­(subvertebral)
venous sinus is located on the midline of the
cranial aspect of the­­carapace, in the region
where the ­cervical vertebrae join the shell. Several
­vessels form a venous sinus at this ­location,
including the ­common ­intercostal ­vessels and
the caudal ­cervical branch of the external jugular
veins. An increased risk of lymph contamination
exists at this ­sampling site3.
Procedure. The animal is placed in ­ventral
recumbency, preferably at the edge of a
table, and the forelimbs are restrained
(Fig. 2). The person performing the
­v enipuncture either extends the head
and bends the neck ventrally (Fig. 2a)
or pushes the head back into the shell.
A piece of gauze or a cotton ball placed
over the head and face can be used to pre-
vent the person obtaining the sample from
getting bitten during the venipuncture
(Fig. 2b). In medium to large chelonians, a
1.5- to 2-inch hypodermic needle should
be used in order to reach the subcarapacial
304 Volume 37, No. 7 | JULY 2008
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space. The needle is directed dorsocaudally
and inserted on midline at the point where
the skin attaches to the ventral aspect of the
­cranial carapace. While mild negative ­pressure
is maintained, the needle is advanced dorsally
until either blood is aspirated or the carapace
is reached. If no blood is aspirated, the ­needle
is then slightly withdrawn and redirected
until blood appears in the syringe hub.
Brachial vein
The brachial vein (venous plexus) is a
­suitable alternative for obtaining ­diagnostic
blood samples in larger species (>1 kg),
for which jugular venipuncture is more
­invasive and stressful, and which are ­typically
­uncooperative4,5. Lymph contamination may
occur, but several studies have shown a lower
risk compared to other sampling sites4,5.
Anatomy. The brachial vein is a plexus of
vessels at the ventromedial (flexor) surface
of the radiohumeral joint (elbow)1.
Procedure. The animal is placed in ventral
recumbency, and the foreleg is extended by
either an assistant or the person ­performing
the venipuncture. The needle is ­inserted
perpendicular to the humerus in the groove
above the prominent biceps brachii ­tendon on
the ventromedial aspect of the ­radiohumeral
joint. The needle is advanced toward the joint
until blood is aspirated (Fig. 3a)1,2.
Other sites
The femoral vein can be considered in cases
where the other recommended sites fail to
yield a diagnostic sample. The ­femoral vein
runs on the ventral surface of the femur
(Fig. 3b). The dorsal coccygeal vein is ­located
at the ­dorsal ­midline of the tail (Fig. 3c).
Because the risk of lymph ­contamination at
this site is very high, it is not recommended
for routine sampling unless venous access
cannot be obtained otherwise3.
COMPLICATIONS
Hematoma formation after blood ­collection
may occur due to improper venipuncture
technique or an underling blood ­clotting
disorder. Whenever possible, manual
­compression should be done ­immediately
after withdrawal of the needle to ­minimize
h e m a t o m a f o r m a t i o n . Ia t ro g e n i c ,
­temporary swelling and erythema of the
neck can occur when jugular ­venipuncture
is done, due to excessive manual force
­during ­extension and restraint of the neck.
a
b c
FIGURE 3 | Blood collection from (a) brachial
vein of the left forelimb in an African spurred
tortoise (Geochelone sulcata), (b) femoral
vein of the left hindlimb in a leopard tortoise
(Geochelone pardalis) and (c) dorsal coccygeal vein
in a red-eared slider (Trachemys scripta elegans).
Lymph dilution
Lymph dilution or contamination is a
­c ommon complication during blood
­collection in chelonians. ­Lymph-diluted
blood samples will result in ­e rroneous
hematological, plasma biochemical and
serological results5–7. Lymph dilution can
occur at any sampling site, but samples
­collected from the jugular vein are least
likely to be lymph-diluted.
1. Heard, D.J., Harr, K.E. & Wellehan, J.F.X.
Diagnostic sampling and laboratory tests. in BSAVA
Manual of Reptiles (eds. Girling, S.J. & Raiti, P.)
71–86 (BSAVA, Gloucester, UK, 2004).
2. Murray, M.J. Reptilian blood sampling and artifact
considerations. in Laboratory Medicine: Avian
and Exotic Pets. (ed. Fudge, A.M.) 185–192 (W.B.
Saunders, Philadelphia, 2000).
3. Barrows, M., McArthur, S. & Wilkinson, R.
Diagnosis. in Medicine and Surgery of Tortoises and
Turtles (eds. McArthur, S., Wilkinson, R. & Meyer,
J.) 132–140 (Blackwell, Ames, IA, 2004).
4. Zaias, J. et al. Biochemical and hematologic
values for 18 clinically healthy radiated tortoises
(Geochelone radiata) on St Catherines Island,
Georgia. Vet. Clin. Pathol. 35, 321–325 (2006).
5. Lopez-Olvera, J.R. et al. Effect of venipuncture site
on hematologic and serum biochemical parameters
in marginated tortoise (Testudo marginata).
J. Wildlife Dis. 39, 830–836 (2003).
6. Gottdenker, N.L. & Jacobson, E.R. Effect of
venipuncture sites on hematologic and clinical
biochemical values in desert tortoises (Gopherus
agassizii). Am. J. Vet. Res. 56, 19–21 (1995).
7. Berry, K.H. et al. A comparison of lymph and
plasma sample results from ELISA tests for
Mycoplasma agassizii in desert tortoise. in 30th
Annual Meeting and Symposium of the Desert
Tortoise Council. (Tucson, AZ, 2005).
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