CSF Analysis and Their Interpretations in Animals

Analysis of CSF samples &
their Interpretations
Submitted by –
Dr. Ramesh Kumar
M.V.Sc. Scholar
Department of VGO
C.V.A.S.(RAJUVAS), Bikaner
Cerebro-Spinal Fluid
 It is a secretion of choroid plexus and ependymal cells.

 Fluid from lateral ventricles flows through the interventricular foramina
(Monro) to the 3rd ventricle and then to the 4th ventricle by the narrow
aqueduct of sylvius.
 From here it reaches 4th ventricle by two lateral foramina (Luschka).
 The 4th ventricles opens to the central canal of spinal cord. The CSF
circulates in the subarachnoid space (between the pia and arachnoid)
and at some sites it enlarges such as cisternal magna, thus this location
is used for puncture to obtain CSF.
Function of CSF
1. It is a buffer fluid which acts as cushion for the brain and spinal
cord.
2. It helps in exchange of metabolites between blood and brain.
Indications for CSF analysis
1. When a CNS disorder is suspected.

2. Repeated CSF analysis can be helpful to evaluate response to
treatment.
Collection of CSF
 In dog, the preferred site for

obtaining CSF is the Cisterna magna
at atlanto- occipital articulation.
 Same site is also useful for horses,
cattle, sheep and goats.
 In these animals, sublumbar
puncture can also be used.
Atlanto- Occipital puncture
 The CSF can be obtained by atlanto-occipital puncture in lateral recumbency during
general anesthesia in dogs, cats and equine and done in standing position or in
lateral recumbency under sedation (xylazine hydrochloride at 0.05 mg/ kg,IV) and
local analgesia (2% lignocaine hydrochlo ride) in ruminants with following steps:
 Restrain the animal.
 The head is flexed at the atlanto-occipital joint approximately 90º (ensure that airway
remain patent during flexion), so that the long axis of the head is perpendicular to
spine.
 Clip, clean, and sterilize the selected area.
 Puncture in the midline along an imaginary line drawn between the cranial borders of
the atlas wings.
 Use a 3-4 inch long and 16 G spinal needle with a stylet and insert it slowly at the
cranial edge of the wings of atlas. Direction of the needle should be parallel to the
long axis of the head.
 When the needle enters subarachnoidal space,loss in resistance is felt.
 As the needle enters into atlanto-occipital joint, remove the stylet so that the CSF
flows out and collect approximately 1 to 2 ml of CSF.
Lumbo-Sacral Puncture
 The collection is done under local analgesia (2%lignocaine hydrochloride)
in the standing position.
 Presurgical and aseptic precautions are taken and the depression between
the dorsal process of last lumbar vertebra and cephalic end of median
crest of sacrum is palpated.
 Needle is passed at this site. Insert the needle vertically, then slightly
oblique by applying gradual pressure in forward and backward directions.
 As the needle enters in subarachnoid space comparatively less resistance
is felt.
 Animal must be tied firmly to avoid damage to the spinal cord. CSF
collection is done by removing the stylet, apply a syringe and suck the
fluid.
Analysis of CSF
 Physical Examination
 Chemical Examination
 Cytology
 Bacteriological Examination
Physical Examination
Colour
 Normal CSF is clear and colourless.
 Slight pink or reddish colour indicates hemorrhage.
 Centrifuge the CSF. If the supernatant is clear, it indicates a blood
vessel puncture in contrast to yellow colour in old haemorrhage, from
progressive brain disease such as inflammation or neoplasia.
 A gray or green colour indicate suppuration.
Turbidity
 Turbidity is due to presence of a high cell count (pleocytosis).
 Neutrophils are seen in bacterial encephalitis, abscess, immune
mediated meningitis and hemorrhage.
 Increased number of monocytes are seen in viral encephalitis, fungal
infections, uremia, chronic conditions and post vaccinal reactions.
Coagulation
 Normal CSF does not coagulate but inflammatory conditions such as
acute suppurative meningitis, produce coagulation.
Specific Gravity
 It is 1.003 to 1.008.
Reaction
 It is similar to blood i.e. alkaline.
Chemical Examination
Protein
 Normally, the protein level is less than 25 mg/dl.
 Increase in total protein levels indicate increase in globulin levels.
 For this shake the tube of CSF. Foam is formed which lasts for 5 minute
or more when protein is increased.
 Similarly 3 ml of 3% sulfosalicyclic acid added to 1 ml of CSF produces
turbidity after 5 minutes when protein is increased.
Glucose
 Normal CSF has glucose 40-110 mg/dl.
 Glucose levels are decreased in acute pyogenic meningitis,
hypoglycemia, and metastatic carcinoma to the meninges.
 High glucose level is seen in hyperglycemia.
 Little increase or no change occur in viral encephalitis, brain abscess
and brain tumors.
Chloride:
 The levels are normally higher in CSF than in blood.
 Normal CSF values in domestic animals ranges between 650-850 mg/dl.
 Lower values are seen in pyogenic meningitis, protracted vomiting,
advanced pneumonia, hypochloremia, while normally higher values of
chloride in CSF are recorded than in serum.
Sodium:
 The concentration of sodium in CSF is similar to the value in serum.
 CSF concentration of sodium>160 mEq/L is considered diagnostic for salt
poisoning.
Calcium:
 Normally the calcium is lower in CSF than in serum.
 Increased level of protein bound calcium in CSF indicates disturbance in
blood brain barrier.
Cholesterol:
 Haemorrhages in the CNS, tumors, meningitis and brain abscess lead to
an increase in cholesterol content. Usually normal cholesterol level is
very low and values recorded in Horse: 0.36 – 0.55 mg/dl, and Goat: 0.51
mg/dl.
Enzymes
 Lactic dehydrogenase (LDH), transaminases (AST, ALT), Creatine
phosphokinase (CPK) are usually increased in inflammatory conditions of
central nervous system e.g. LDH levels in bacterial meningitsis, metastatic
carcinoma and transaminase levels in purulent meningitis and canine
distemper.
Electrolyte
Concentration
Measurement techniques are same as for blood.
Sodium, potassium, chloride and magnesium can
be measured but are of very little practical value.
Cytology
Cell Count
Undiluted:
 As cells disintegrate, rapidly count within 20 minutes after collection, using a Neubauer's hemocytometer chamber.
 Each side of the hemocytometer chamber is charged with one drop of fresh undiluted CSF.
 Cells are allowed to settle for 10 minutes.
 Calculate average of two sides and multiply by 1.1 to produce the number of cells per microliter.
With diluting fluid:

 Draw fluid in leukocyte diluting pipette up to mark 1 and fill the pipette with CSF up to mark 11.
 Mix, discard few drops and fill each side of hemocytometer chamber.
 Wait for few minutes and count the cell in entire ruled area on both sides of the counting chambers i.e. 18 one-mm square.
 Multiply by 0.6 to obtain number of cells per mcroliter.
Diluting Fluid:
 Glacial acetic acid 10ml
 Distilled water 90ml
 Crystal violet 0.1 g
 Add 2 drops of phenol (5%) as preservative.
Interpretations
 The normal cell count is less than 20/microlitre. Marked increase is seen
in acute pyogenic meningitis (100 to 500/microliter) while moderate
increase is seen in encephalitis. Mild increase occurs in viral infections,
degenerative diseases and neoplasia.
Differential Cell Count of CSF
 If total cell count is less than 500/microliter, centrifuge the CSF,
discard supernatant, add one or two drops of serum and make a smear
from sediment.
 Air dry and stain as for blood smear.
 Neutrophils are not present in CSF. When present, indicates pyogenic
or bacterial infection.
 Lymphocytes are increased in viral infections, fungal infections,
chronic infections and toxic conditions.
Bacteriological Examination
 It is carried out when the CSF cell count and protein contents are high.
 When CSF is turbid, gram stain and new methylene stain (NMB) should
be made.
 Culturethe CSF on blood agar, in thioglycollate culture medium or in
sabourad dextrose agar depending upon findings in the direct smear.
 When the CSF is not turbid, centrifuge it before staining and culture
the sediment.
 NMB and India ink will show capsule of cryptococcal organisms.
CSF reference values
Thank you 😊

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CSF Analysis and Their Interpretations in Animals

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Analysis of CSF samples &

 It is a secretion of choroid plexus and ependymal cells.

1. When a CNS disorder is suspected.

 In dog, the preferred site for

With diluting fluid:

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