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Lab D > 8. Ruminal fluid > Flashcards

8. Ruminal fluid Flashcards

1
Q

Indications for ruminal fluid examination

A
  • Diagnosis of ruminal diseases

- Evaluation of ruminal fluid before use in therapeutic transfusion (transfaunation).

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2
Q

Methods to obtain ruminal fluid

A

By orogastric tube

By ruminal puncture

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3
Q

How should transportation of ruminal fluid be for long distance be done?

A

Must be done in double jacket container.

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4
Q

Estimation of chloride and ammonia conc. can be delayed how long?

A

Up to 9 hrs in room temperature and up to 24 hrs in refrigerator

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5
Q

What is ruminal fluid properties influenced by?

A
  • The composition of the diet
  • The circumstances of the feeding and sampling
  • The time interval from feeding or drinking
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6
Q

Physical examination of ruminal fluid - what do we examine?

A
  • Odor
  • Color
  • Consistency
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7
Q

Odor of ruminal fluid: normal and abnormal

A 
Normal: Aromatic odor
Abnormal:
-Ammonia smell (urea poisoning)
-Moldy rotting (protein putrefaction)
-Acidic or sour odor (excess lactic acid/grain overfeeding)
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8
Q

Color of ruminal fluid: normal and abnormal

A

Normal:
 Olive to brownish green (hay ration)
 Deeper green color (green ration)
 Yellowish brown color (grain or silage ration)
Abnormal:
 Milky grey (grain overfeeding, lactic acidosis)
 Darker greenish or brownish (ruminal stasis/decomposition)
 Grey with clots of milk (calves with abomasal reflux)

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9
Q

Consistency of ruminal fluid: normal and abnormal

A

Normal:
 Slightly viscous
Abnormal:
 Increased viscosity (saliva contamination)
 Decreased viscosity, watery with few feed particles (inactive bacteria or protozoa)
 Excess frothy, stable bubbles (frothy bloat/ vagus indigestion)

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10
Q

Sedimentation time or sedimentation/flotation test:

  1. Which particles sink/float?
  2. Normal range is?
  3. What does sedimentation of fine particles with subsequent flotation state?
  4. What does rapid sedimentation state with little to no flotation?
  5. What does stable froth presence indicate?
A
  1. Smaller particles sink, larger particles float on the bubbles of fermentation.
  2. 4–8 minutes
  3. Very active fluid
  4. Inactive fluid, due to lack of fermentative gases
  5. frothy bloat or some types of vagal indigestions in the Hoflund disease or in case with treatment with sympathicomimethics (acetylcholine) used for the treatment of ruminal atonia.
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11
Q

Biochemical examination - what do we examine?

A
  • pH
  • VFA´s
  • The reducing ability of the anaerobic rumen flora
  • Ruminal flora and fauna
  • Rumen fluid ammonia
  • Rumen fluid chloride
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12
Q

When should we measure pH of ruminal fluid?

A

Best measured 2–4 hours after feeding concentrates or 4–8 hours after feeding a total mixed ration.

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13
Q

Which samples have lower pH?

A

Samples collected by rumenocentesis have lower pH values than those collected by oral routes.

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14
Q

Normal and abnormal pH of ruminal fluid

A 
Normal pH: bw 6.3–7 (slightly acidic)
-cows fed grain can have slightly lower
-those fed hay or green slightly higher pH
Abnormal pH: 
1. Elevated pH (Rumen alkalosis)
-Simple indigestion or reduced feed intake for greater than 2 days
-Urea indigestion
-Putrefaction of ruminal content from prolonged rumen stasis
- Saliva contamination
2. Lowered pH 5.5.-6 (Rumen acidosis)
-Grain overfeeding
-Chronic ruminal acidosis
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15
Q

Which are the principles VFA ?

A

Acetic (major), propionic and butyric acids.

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16
Q

The average ratio of VFA`s at the pH 6-7?

A

The average ratio at the pH 6-7 is 60-65% acetic acid, 20-25% of propionic acid and 10-15 mol% of butyric acid and 5 mol% other acids.

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17
Q
  1. If the crude fiber content is around 20%, what is the acetic acid:propionic acid ratio?
A

4:1

18
Q

What does a decrease in the VFA content indicates?

A

A diet low in fiber or insufficient intake, and decreased activity of the microbial organisms.

19
Q

High grain diet does what with pH of ruminal fluid?

A

Decreases the pH of the ruminal fluid.

20
Q

How can we measure VFA?

A

With high performance liquid gas chromatography (HPLC).

21
Q

The concentration of VFA in the rumen is?

When is it highest?

A

Bw 80-120 mmol/l.

It is the highest around 3 to 5 hours after feeding

22
Q

Tests for the reducing ability of the anaerobic rumen flora: what organism has the most important role, and why?

A

Bacteria; Their dehydrogenating activity can be shown with different color reagent.

23
Q

How can be see the redox processes from bacteria on rumen flora?

A

Some of them will be reduced to colorless forms (methylene blue); others from colorless oxidized form will
be reduced to colored reagent (triphenyltetrazolium-chloride, TTC-test).

24
Q

How is reduction in rumen flora mostly done?

A

Mostly by the organisms which play a role in carbohydrate fermentation. In diet rich in starch and during ruminal acidosis the redox potential is increased. When the ruminal flora is destroyed (e.g. putrefaction), the redox potential is reduced.

25
Q

Tests for the reducing ability of the anaerobic rumen flora

A
  • Nitrite reduction test: ruminal bacteria’s reduce nitrites and nitrates.
  • Methylene blue reduction test
26
Q

Nitrite reduction test

  1. Solution used?
  2. Interpretation?
  3. When is the reduction more rapid/slower?
A
  1. Potassium nitrate (KNO2), then Griess-Ilosvay reagent
  2. Interpretation:
    - In the presence of nitrite the color will be pink. In a healthy ruminal fluid after 10 minutes normally there is no nitrite.
  3. Reduction is more rapid when cattle are fed green fodder or have ruminal decomposition or bloat, and slower when a deficient ration is fed or inapetence.
27
Q

Methylene blue reduction test

1. Interpretation?

A
  1. Interpretation:
    -Normal: 3–6 minutes; concentrate diets result in faster times, forage diets result in longer time of range
    -Prolonged: Discoloration which takes longer than 10–15 minutes indicates inadequate anaerobic bacterial
    population, rumen acidosis or indigestible roughage.
28
Q

TTC test

A

Triphenyltetrazolium-chloride. It is performed in the same way as methylene blue reduction test.
-In this case we expect a red coloration after incubation, after 10-15 min.

29
Q

Which microbes can we find in rumen? How much do they digest of the digestible dry matter?

A

Bacteria, protozoa and fungi. Bacteria and protozoa are the predominant microbes. They digest about 70% to 80% of the digestible dry matter in the rumen.

30
Q

Which protozoa can we find in the rumen?

A

Both ciliates and flagellates are present in the rumen, but only the former are of physiological importance by virtue of their number and mass.

31
Q

In which conditions are ruminal ciliates living under?

A

Strict anaerobic conditions.

32
Q

What are the tasks of protozoa in the rumen?

A
  • breaking down soluble sugars and various polysaccharides or storing them
  • prevent rapid fall in ruminal pH by utilizing some starch
    that otherwise would be broken down by bacteria
  • their active movement help at the mixing of the ruminal content
33
Q

Microscopic examination of protozoa

A

Should see three groups of protozoa: small, medium and large and they should be rapidly moving.

34
Q

When is the motility of protozoa decreased?

A

When the environmental changes.
In case of chronic processes, they do not disappear completely, but their number and motility is reduced. Always the biggest sized protozoa will be
affected first.
Errors: if the temperature of the ruminal fluid is decreased, the slide is cold, the motility of the protozoa will be reduced.

35
Q

Bacteria

  1. Why are they important?
  2. Conc. in fluid rumen
  3. When is there less bacteria present?
  4. How can they be examined?
  5. What can gram stain reveal?
  6. Which bacterias are predominant in rumen fluid?
  7. What does lactic acidosis result in?
  8. Which bacterias are predominant in a diet rich in fiber?
  9. Which bacterias are predominant in a diet rich in starch?
  10. In case of ruminal putrefaction, which bacterias are present in high conc.?
A
  1. They are vital for the fermentation processes of the forestomach.
  2. Bw 107 and 1012 per ml of fluid rumen.
  3. When the ration is rich in cellulose (crude fibre) compared with ratio rich in starch.
  4. Staining or culturing. Staining can be performed with Giemsa, Gram or PAS staining.
  5. Gram stain reveals the populations of bacteria present. 6. Gram negative bacteria.
  6. In a more homogenous, predominantly gram-positive population.
  7. Gram negatives
  8. Gram positives
  9. Proteus, Clostridium and Pseudomonas
36
Q
  1. Which fungi are mostly present in the rumen?
  2. What do they contribute to?
  3. How can they be revealed?
  4. What happens with the fungi in case of acidosis or putrefaction?
A
  1. Yeast
  2. Carbohydrate (mostly cellulose) digestion.
  3. By Giemsa, PAS staining or culturing.
  4. They disappear completely from the rumen, or they multiple on the place of bacteria.
37
Q

Rumen fluid ammonia:

  1. How is ammonia synthesized in rumen fluid?
  2. How is it absorbed?
  3. What is the function of ammonia?
  4. How can it be measured?
  5. What is the difficulty in measuring ammonia?
A
  1. From protein and non-protein nitrogen (NPN) sources by the ruminal bacteria.
  2. Through the ruminal wall and passes further through the portal vein.
  3. Used as energy source or is built up in the bacterial protein.
  4. With a spectrophotometer or with ion selective method.
  5. By the fact that NH3 is highly volatile. On the other hand the NH3 from the environment (stable, smoke) will contaminate the sample. The sample should be immediately used for the measurement, or transported in closed system between 0-2 C°.
38
Q

Rumen fluid ammonia: Interpretation

  1. Normal level
  2. Cause of increased ammonia
  3. Effects of increased ammonia
A
  1. Normal level in rumen is low (6-20 mmol/l).
  2. Incr. ammonia: In case of high dietary protein intake or in energy deficiency.
    -Liver disorders also can cause increased ammonia levels. After calving the liver function is
    reduced because of energy deficiency and fatty infiltration.
  3. Effect: will lead to ruminal alkalosis and in severe cases to ammonia intoxication.
39
Q

Rumen fluid chloride

  1. Conc. in healthy ruminant?
  2. Its main source?
  3. Cause of increaed chloride
  4. What type of samples is used for measurement?
  5. How long can it be stored?
  6. Type of measurement methods?
A
  1. Low: 15-20 mmol/l
  2. Main source: the diet and the saliva which passes into the rumen.
  3. In case of gastric torsion or other cases of pylorus obstruction the hydrogen chloride will pass into the rumen (reflux phenomenon) and can incr. chloride up to 30-100 mmol/l.
  4. In a supernatant of a centrifuged sample.
  5. Relatively stable over time: room temp up to 9 hours and refrigerated for 24 hours
  6. Spectrophotometer or ion selective method.
40
Q

Interpretation of rumen fluid chloride

  1. Normal level
  2. Causes of elevated level
A
  1. Normal level in cattle is < 30 mEq/L (15–20) and in sheep < 15 mEq/L
  2. Elevated level:
    - High salt intake
    - Abomasal displacement
    - Obstruction of intestinal flow (ileus), pyloric stenosis,
    - Abomasal disorders (inflammation, ulcer, hyperacidity)

Lab D (18 decks)

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