Clinical Biochemistry of Domestic Animals (Sixth Edition) - UMK ...

VI. CSF Collection and Analytical Techniques
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recommended for large breed, nonchondrodystrophic dogs
(e.g., German shepherd dogs), whereas L5 to 6 is recommended
for small, chondrodystrophic dogs (e.g., dachshunds)
(Morgan et al., 1987 ). The puncture site chosen should be as
close to the lesion as possible without penetrating the lesion,
or the site should be caudal to the lesion. In animals with
spinal disease, cerebellomedullary fluid is abnormal more
frequently with cervical disease than it is with thoracolumbar
disease, but overall lumbar fluid is abnormal more often
than cerebellomedullary fluid. With intracranial disease,
CSF from both sites is usually abnormal, perhaps because
both sites are caudal to the lesion ( Scott, 1992 ; Thomson
et al., 1989, 1990 ). Occasionally, CSF is collected from both
sites. Although the order of collection (cerebellomedullary
or lumbar CSF collected first) appears not to influence significantly
the analytical results ( Bailey and Higgins, 1985 ),
aspiration from the relatively small lumbar subarachnoid
space is easier if the CSF pressure has not just been lowered
by cerebellomedullary CSF collection.
3 . CSF Collection from the Cerebellomedullary
Cistern
The authors ’ preferred technique for CSF collection from
the cerebellomedullary cistern is to utilize the palpable bony
landmarks that are the closest to the puncture site. These
structures are the vertebral arch of C1 and the external
occipital protuberance. After anesthetic induction and intubation,
the animal is placed in right lateral recumbency, and
padding is placed under the neck to align the dorsal cervical
and cranial midline parallel to the tabletop. The assistant
is instructed to tuck in the animal’s chin (flex the neck) and
push the external occipital protuberance toward the operator.
This procedure flexes the atlantooccipital joint and maximizes
the space between the occipital bone and C1. Asking
the assistant to simply flex the neck seems to produce flexion
of the midcervical area more than the atlantooccipital area.
The clinician faces the dorsal aspect of the patient’s neck,
kneeling on a pad. The external occipital protuberance, the
C2 spinous process, and the C1 vertebral arch are palpated.
The latter structure is located by rolling a fingertip off the
cranial edge of the C2 spinous process and palpating firmly,
feeling for a transverse bony ridge (the C1 vertebral arch).
The C1 vertebral arch can usually be palpated, and if so, the
puncture is made on the midline just in front of the fingertip
palpating the vertebral arch. If C1 is not palpable, the distance
between the cranial edge of the C2 spinous process and
the occipital protuberance is noted, and the puncture is made
on the midline about one-third of that distance cranial to the
cranial edge of the C2 spinous process. In rare cases, neither
C1 nor C2 can be palpated. In this situation, the lateral
edge of each C1 transverse process is palpated and a triangle
from each edge to the occipital protuberance is constructed
visually. The puncture is made on the midline in the center
of that triangle. The needle should be advanced slowly
and the stylet removed regularly. A “ pop ” may be palpated
when the dura mater is punctured with the needle. The clinician
should hold the spinal needle with one hand (to hold
it steady) and remove the stylet with the other hand. The
CSF should be allowed to drip out of the spinal needle into a
tube. A volume of at least 0.5 mLs should be collected for a
full CSF analysis (partial analysis may be done with smaller
volumes). Larger volumes may be collected for other tests
such as culture and sensitivity, polymerase chain reaction
for infectious agents, antigen/antibody testing, immunophenotyping,
and clonality assessment. To collect CSF for culture
and sensitivity testing, aspirate CSF directly from the
spinal needle hub using a needle and syringe.
B . Physical Examination: Clarity, Color,
and Viscosity
After collection, the CSF is examined visually and the color,
clarity, and viscosity are recorded. Normal CSF is clear and
colorless and has essentially the same viscosity as water. For
accurate assessment, the CSF can be compared to the same
amount of distilled water in the same type of container. The
containers can be held against a white, typewritten page to
judge color and clarity, and gently shaken to assess viscosity.
If the CSF appears abnormal, the color and clarity of the
supernatant after centrifugation should be noted.
C . Cytological Analysis
1 . General Techniques
Collection of CSF in a plastic or silicon coated glass tube
is preferred because monocytes will adhere to glass and can
activate in the process ( Fishman, 1992 ). This can result in
erroneous cell counts and also alter morphology. In practical
terms, this is of little consequence in those specimens that
are rapidly processed, but it becomes important as the delay
between collection and processing increases. A complete
cytological examination includes both a total and differential
cell count, as well as thorough morphological assessment.
A differential and thorough morphological assessment
should be done routinely, even on those samples that have
cell counts within normal limits. In our experience, very low
cell counts alone cannot be used as an indicator of normality.
In one study utilizing cytocentrifugation, about 25% of
canine CSF samples with cell counts in the normal range
had abnormalities in cell type or morphology ( Christopher
et al., 1988 ). Abnormalities included the presence of phagocytic
macrophages, increased percentage of neutrophils in
the differential, and the presence of reactive lymphocytes
and plasma cells. Malignant cells have been observed in
samples with normal nucleated cell counts ( Bichsel et al.,
1984b ; Grevel and Machus, 1990 ). CSF samples should be
processed as soon as possible after collection. Cells degenerate
quickly in CSF ( Chrisman, 1992 ; Fishman, 1992 ; Fry