EXAMINATION OF

RUMEN FLUID
Examination of rumen fluid were important to
A-obtained an accurate diagnosis of diseases of the
rumen.

B-Evaluation of ruminal fluid before use in

therapeutic transfusion
• Sample (s) which must be taken…..

1- should be evaluated as soon as possible after

collection to minimize effects of cooling and air
exposure on protozoal activity and pH.
2- Transportation of rumen fluid for long distance

must be done through container.

3- Samples should be stored at room temperature

(20-22)˚c and examined within 9 hours of
collection,

Whereas samples kept in a refrigerator (4-5)˚c

should be examined within 24 hours
Examination included …..
Physical Examination:

The Color:

Normal color varies depending on the nature of feed:

• Olive to brownish green color: hay ration.

• Deep green color: green ration.

• Yellowish brown color: grain or silage.

• Gray color: fodder beet.

• Abnormal color may be:

• Milky gray color: grain overfeeding.

• Greenish-black color: prolonged stasis of the

rumen, and/or decomposition of rumen contents.

• Gray with clot of milk: calves with abomasal

reflex or esophageal groove failure

The Odor:
• Normal odor is aromatic. Abnormal may be:
o Ammonia smell: urea poisoning.
o Mouldy rotting odor: protein putrefaction.
o Intensely sour odor: excess lactic acid
production from grain or carbohydrates
overfeeding.
The Consistency:
•Normal consistency is slightly viscous.
•Abnormal consistency may be:
o Watery: in active bacteria and protozoa.
o Excess frothy: frothy bloat from primary
tympany or vagus indigestion
Chemical Examination:
The pH:
The pH may be measured by using indicator paper or,
an electrical pH meter.
Normal pH ranges between 6-7 in animal on a mostly
forage diet, but is lower 5.5-6.5 in animals fed mostly
grain.
•Elevated pH (Rumen alkalosis):
o Simple indigestion.
o Urea indigestion.
o Putrefaction of rumen ingestion.
o Feeding on indigestible roughage.
•Lowered pH (Rumen acidosis):
o Chronic rumen acidosis (pH 5-5.5).
o Abomasal reflex from abomasal disease.
o Vagal indigestion.
o Intestinal obstruction
The Sedimentation activity test(Evaluation of microfloral
activity)
The test were provides a rapid evaluation of microfloral
activity.
• a sample of rumen fluid inserted into a test tube and

let to stand. Measure the time needed for complete

sedimentation, which is reflected the sediment activity
time.
• Normal time is 4-8 minutes.

• Abnormal time may be:

• Very rapid sedimentation with no floatation

occurs in rumen acidosis, prolonged anorexia,

inactive microflora from indigestible roughage.
• No sedimentation and floatation in frothy bloat,

some cases of vagal indigestion.

The Methylene blue reduction test:

The test reflect the anaerobic fermentation metabolism

of bacterial population.
• Mix 20 ml of rumen content with 1 ml of 0.03%

methylene blue in a test tube and let to stand at room

temperature. then Measure the time needed for color
change of the mixture.
•
• Normal rumen fluid from cattle fed on a hay and

grain diet needs 3 minutes to decolorize leaving a

narrow ring of blue color at the top of
decolorizing mixture.
• Abnormal reduction of time up to 15 minutes

indicates indigestible roughage, anorexia of

several days, or rumen acidosis.
The Cellulose digestion test:
Mix 10 ml of rumen fluid with 0.3 ml of 16% glucose solution in a
test tube. Immerse a thread of pure cellulose (free from
synthetic fiber). The lower end is weighted by a glass bead
• Incubate the tube at 39˚c.

• Record the time for the bead to be dropped free at the bottom

of the tube.
active rumen fluid will digest the cellulose within 48-56 hours. The
test takes a long time and is not very accurate.
The Glucose fermentation test:
The test measures( indirectly) the ability of microflora to
breakdown glucose by measuring the volume of gas formed.
• 0.5 ml of 16% glucose solution is added to 10ml of rumen fluid.

Place the mixture in a fermentation saccharometer

and keep at 39˚c. Read the result after 30 and 60 minutes:
• The normal rate of gas formation is 1-2 ml/hour.

• If the microbial flora is inactive, little or no gas forms.

• In foamy bloat, more gas is formed with a lot of foaming.

Nitrate reduction test:
The test give an idea on activity of microbes that synthesize
nitrogen compounds.
• 10 ml of sieved ruminal fluid is placed into each of 3 test tubes

and 0.2, 0.5, 0.7 ml of 0.025% potassium nitrate solution is added

to 3 tubes
• Put the 3 tubes in water both at 39˚c. Every 5 minutes, one drop

from each tube is placed in the small ceramic plate. To each

drop is added 2 drops of reagent 1(sulphanilic acid &acetic acid)
and 2 drops of reagent 2(alpha-naphthylamine concentration
acetic acid) Observe the change of color.
• Sample that contain nitrates are colored red.

• Rumen fluid of cattle fed a mixed ration will not change in

color after 5-10 minutes in tube 1 and 20 minutes in tube

II, and 30
minutes in tube III.
• Reduction is more rapid when cattle are green fodder or

have ruminal decomposition or bloat.

• Reduction is more slower when a deficient ration is fed

or when the animal lacks the appetite

Rumen fluid chloride

Measured in a supernatant of a centrifuged sample

(rumen fluid sample )
• It Measured by chloride meter.

• Normal level is :30 mEq/l

• -Elevated level occur in:

• * Abomasal disease.

• * Obstruction of intestinal flow..\rumen.mp4

Examination of rumen Protozoa
The protozoa of the ruminal fluid include both
ciliates and flagellates, but only the ciliates are of physiological
importance
Quantitative examination:
Dilute 1 ml of rumen fluid with 15 ml saline solution and 5 ml Lugol's
iodine solution. Shake gently. Spread 0.1 ml of the mixture on glass
slide in an area under cover glass of the 22 × 50 mm.
• Counting is carried out using low power(10x). The field area of that
lens is one square millimetre.
• Count 30 fields in the slide, Which represent the protozoal count
over one square millimetre area of the
field.
• Multiply the average by 1100 to the protozoal count in 0.1 mm of
the diluent fluid which represent 0.02 ml of the original sample.
• Multiply the obtained figure by 50 to obtain total protozoal per ml.
Examination of the Bacteria

Air-dried smear of rumen fluid is

stained by Gram's method, or other staining methods

Interprettation of the smear are:

• A diet rich in starch produces a more uniform picture,

with Gram-negative cocci, short and long rods, and a

relatively high proportion of Gram-positive cocci and
rods.