Rumen Fermentation

Rumen Fermentation

Worlds largest commercial fermentation space

100 billion liters or rumen volume in domestic animals 1010 to 1012 cells/mL 200 liters (50 gallons) in one cow

Ruminants

Continuous culture fermenters

Input and output

Lignocellulosic substrates digested

Cellulase complex Hemicellulases Lysozyme Nitrogen capture (NPN)

8 x 1015 mouths to feed

Because of these microbial enzymes, ruminants can utilize feedstuffs that provide little to no nutritional benefit to nonruminants

4 Steps of Rumination

Regurgitation

reverse peristalsis carries food to mouth

Remastication

liquid squeezed from bolus and swallowed bolus chewed

adding more saliva swallowing bolus and liquid

Reinsalivation

Redeglution

Rumination

Allows animal to forage and eat food rapidly, and then store for later digestion Reduces particle size only small particles leave reticulorumen Increases surface area for microbial attachment and digestion/fermentation Breaks down impervious plant walls Further stimulation of saliva flow (buffer rumen)

Rumination Time

Average times for a grazing animal

Eating 8 hours Ruminating 8 hours Resting 8 hours Reducing forage:concentrate decreases rumination Reducing particle size of forage decreases time spent ruminating

Ruminating time is quite variable

Mechanism of Rumination: Regurgitation

Stimulus digesta in fiber mat scratching surface near cardiac sphincter Contraction of the reticulum forces digesta to cardia Animal inhales with epiglottis closed to produce a vacuum Cardia sphincter opens and esophagus dilates

Rapid reverse peristalsis moves digesta to mouth

Negative pressure (vacuum) sucks digesta into esophagus

Mechanism of Rumination: Remastication, Reinsalivation, and Redeglutition

Bolus is rechewed

Digesta reinsalivated

Chewing is slower and more deliberate than during initial eating phase Parotid glands secrete more saliva during rumination than eating
Saliva from parotid glands secrete more NaHCO3- than other glands

Reswallowing

After reswallowing, the rumen contracts to move swallowed bolus into the rumen

Reducing Particle Size of Ingested Feeds

Chewing during eating (minimal)

Preparation for swallowing Release soluble constituents Damage plant tissues for microbial attachment Decrease particle size for passage Damage plant tissues for microbial attachment

Chewing during rumination (extensive)

Microbial digestion Reticuloruminal contractions

Rumen Contractions

Inoculate incoming feed with microbes Mix contents

Minimize effects of stratification Move fermentation products (VFAs) to rumen wall

Particle sorting and passage of small particles to omasum Rumination Eructation of fermentation gases

Rumen Contractions

Feeding increases frequency and amplitude of contractions Feeding a finely ground forage reduces number and intensity of contractions

Metabolic problems

Requires 2-6 weeks to adapt

Hardware disease, hypocalcemia, or hyperglycemia will inhibit ruminal contractions

Need for Eructation

Peak gas production occurs 30 min to 2 hr postfeeding (12-27 liters/min)

Composition of rumen gas

__Gas__ CO2 CH4 (variable) N2 O2 (at wall) H2 H2 S _%__ 65.35 27.76 7.00 .56 .18 .01

Average is 1-2 liters/min

Approximately 30% of CO2 produced in rumen is absorbed into blood and removed through the lungs Only 20% of the CH4 is removed through the lungs

Control of Eructation

Stimulus

Gaseous distension of the reticulum and rumen Esophagus dilates & animal belches

12-30 L per minute for cattle 3-17 times per minute

Inhibition

Presence of digesta near the cardiac sphincter

Affects all three sphincters Protective mechanism to prevent digesta from entering lungs

Epinephrine Histamine
Ruminal pressures will increase to 45 to 100 mm Hg Stable froth or foam formed in rumen

Inhibition of eructation will cause the animals to bloat

Why Worry about Rumen Microbes?

Microbes make ruminants less efficient!!
Aerobic fermentation:
Glucose + O2 ATP + CO2 + H2O

Anaerobic fermentation:
Glucose acetic acid + propionic acid + butyric acid + CO2 + H2O + CH4 + Heat

Feed the Microbes, Let the Microbes Feed the Ruminant!

Feed In
VFA Microbial Protein Vitamins

The nutrients presented to the animal after ruminal fermentation are very different than those entering the rumen as feed

Rumen Digestion and Fermentation

CO2 VFA Microbial cells NH3 CH4 Heat Long-chain fatty acids H2S

Degradable Feed

Rumen microbes

Rumen Microorganisms Nutritional Requirements

CO2 Energy

End products from digestion of structural carbohydrates fermentation of sugars Ammonia (majority of nitrogen needs) Amino acids (cellulolytic bacteria) Co, S, P, Na, K, Ca, Mg, Mn, Fe, Zn, Mo, Se None required in mixed cultures

Nitrogen

Minerals

Vitamins

Symbiotic Relationship

Microbes provide to the ruminant

Digestion of cellulose and hemicellulose Provision of high quality protein Production of VFA Provision of B vitamins Detoxification of toxic compounds

Symbiotic Relationship

Microbes provide to the ruminant

Digestion of cellulose and hemicellulose

Cellulases are all of microbial origin Without microbes, ruminants would not be able to use forage crops such as pasture, hay or silage

Symbiotic Relationship

Microbes provide to the ruminant

Provision of high quality protein

50-80% of absorbed N is from microbes

Improved microbial efficiency will provide more microbial protein Can get over 3 kg of microbial protein per day Amino acid pattern is very similar to that required by the ruminant animal

High biological value protein source

Symbiotic Relationship

Microbes provide to the ruminant

Microbes as a feed source

Bacteria and protozoa washed out of the rumen to omasum and into the abomasum

Acidic environment kills microorganisms Digested and absorbed the same as any other feed source in stomach and small intestine Provide amino acids and some energy

Symbiotic Relationship
Microbes provide to the ruminant Energy!!!
VFA Microbial cells 70% 10%

Digestible unfermented feed 20%

No glucose available for the ruminant

Concentration of VFA in rumen = 50 to 125 uM/ml

Symbiotic Relationship

Microbes provide to the ruminant

Provision of B vitamins

Meets the ruminants requirements under most conditions

Some supplementation, such as niacin, may be beneficial in early lactation dairy cows

Symbiotic Relationship

Microbes provide to the ruminant

Detoxification of toxic compounds

Example:

Mimosine in Leucaena causes problems poor growth, reproduction and hair loss Hawaiian ruminants, but not those from Australia, have microbes that degrade mimosine so Leucaena could be fed Transferred rumen fluid to Australia Inoculated rumen Fed Leucaena safely to Australian ruminants!

Symbiotic Relationship

Ruminants provide to microbes

Housing Garbage removal Nutrients Optimal environment for growth

Symbiotic Relationship

Ruminants provide to microbes

Housing

Reliable heat (39 2C) Fluid environment (free water intake)

85 to 90% water

Guaranteed for 18 to 96 hours depending on diet and type of animal

Straw-fed water buffalo longest rumen residence time for microbes Small selective browsers (mouse deer or duiker) shortest residence time for microbes

Symbiotic Relationship

Ruminants provide to microbes

Garbage removal

Absorption of VFA

Energy to ruminant CO2 and CH4

Eructation

Passage of indigestible residue and microbes to lower GI tract

Rumen mixing to separate and settle small particles

Symbiotic Relationship

Ruminants provide to microbes

Nutrients

Substrates come from feedstuffs that animal consumes Saliva provides urea (N source for bacteria)

Symbiotic Relationship

Ruminants provide to microbes

Optimal environment for growth

Reduced environment (little to no oxygen)

Strict anaerobic microbes in rumen interior Functional anaerobes near rumen wall Saliva contains bicarbonate and phosphate buffers Cows produce up to 50 gallons of saliva daily Continuously secreted More added during eating and rumination Cow ruminates 10-12 hours/day Decreases in particle size of forage reduce need for rumination, decrease chewing time, decrease saliva production, and rumen pH plummets

pH 6.0 to 7.0

Symbiotic Relationship

Ruminants provide to microbes

Optimal environment (pH)

If pH 5.7 rather than 6.5

50% less microbial synthesis Cellulolytic bacteria function best at pH ~6.8 Rate of structural carbohydrate use is decreased Amylolytic bacteria function best at pH ~5.8 More lactate and less acetate is produced Further downward pH spiral

In concentrate selectors (like deer), parotid salivary glands are 0.3% of body weight

Bacteria and pH Tolerance

Species Type pH

Ruminococcus flavefaciens fiber 6.15 Fibrobacter succinogenes fiber 6 Megasphaera elsdenii lactate user 4.9 Streptococcus bovis lactate producer 4.55

Microbes
% of mass Generation No./mL interval Bacteria Protozoa 60-90 10-40 20 min 8-36 h 25-80 billion 200-500 thousand

Fungi

5-10

24 h

minimal

Rumen Microbes

Bacteria

>200 species with many subspecies

25 species at concentrations >107/mL

1010 to 1012 cells/mL 99.5% obligate anaerobes

Environmental Niches for Bacteria

Groups of bacteria in the rumen
Free-living in the liquid phase Loosely associated with feed particles Firmly adhered to feed particles Associated with rumen epithelium Attached to surface of protozoa and fungi

Benefits of Bacterial Attachment

Allows bacteria to colonize the digestible surface of
feed particles
Brings enzymes (from microbes) and substrate (from feedstuff) together Protects microbial enzymes from proteases in the rumen

If attachment prevented or reduced, digestion of cellulose greatly reduced

Retention time of microbes in the rumen is increased to prolong digestion Reduces predatory activity of protozoa Over-feeding fat to ruminants can coat forages, reducing bacterial attachment

Rumen Microbes

Protozoa

Large (20-200 microns) unicellular organisms Ingest bacteria and feed particles Engulf feed particles and digest carbohydrates, proteins and fats Numbers affected by diet

Entodinium (Rumen Protozoa)

Rumen Microbes

Fungi

Known only for about 20 years Numbers usually low Digest recalcitrant fiber

Bacterial Populations

Cellulolytic bacteria (fiber digesters)

digest cellulose require pH 6-7 utilize N in form of NH3 require S for synthesis of sulfur-containing amino acids (cysteine and methionine) produce acetate, propionate, little butyrate, CO2 predominate from roughage diets

Microbial Populations

Amylolytic bacteria (starch, sugar digesters)

digest starch require pH 5-6 utilize N as NH3 or peptides produce propionate, butyrate and lactate predominate from grain diets rapid change to grain diet causes lactic acidosis (rapidly decreases pH)

Microbial Populations

Methane-producing bacteria

produce methane (CH4) utilized by microbes for energy represent loss of energy to animal released by eructation

Location of Microbes
Gas Phase Rumen Wall Fiber Mat

Rumen Fluid

Dietary Factors That Reduce Microbial Growth

Rapid, dramatic ration changes

Takes 3-4 weeks for microbes to stabilize

Restricted amounts of feed Excessive unsaturated fat

Bacteria do not use fat for energy Inhibit fiber digestion and microbial growth Different types of fat have different effects

Dietary Factors That Reduce Microbial Growth

Excessive non-structural carbohydrate

Lowers rumen pH (rumen acidosis)

Slug feeding Feed barley or wheat (rapidly fermented) To prevent acidosis, must balance lactate users and producers

Dietary Factors That Maximize Microbial Growth

Maximum dry matter intake Balanced carbohydrate and protein fractions at the same time

Bacteria need both energy and N for amino acid synthesis

Gradual ration changes Feed available at all times

Maintains stable rumen pH

Rumen Function Overview