NON-GENETIC MANIPULATION

OF RUMEN MICROBES
INTRODUCTION
Increasing human population
Demand for food from plant and animal sources.

Food from animal sources

Ruminants, particularly cattle, buffaloes, sheep
and goats are predominant animal species.

Ruminants are fore gut fermenters

Stomach has 4 distinct compartments: Rumen,
Reticulum, Omasum and Abomasum.
 Myriad of microorganisms are found in the GIT (Rumen)
of ruminant, have true symbiotic relationship with the
host.
These rumen microorganisms are predominantly bacteria,
protozoa and phycomycete fungi
The microbes degrade structural carbohydrate and causes
fermentative digestion of large amounts of fibrous feed
consumed by the ruminants.
By providing a suitable habitat for these microorganisms,
the ruminants are able to utilize the end products of
microbial fermentation to meet their own nutritional need.
Rumen is an open, self-contained ecosystem in which feed
consumed by the ruminant is fermented by rumen
microbes to VFAs and microbial biomass those serve as
source of energy and protein for the host animals
Rumen microbial ecosystem: An efficient
anaerobic fermentation system
Ruminants can digest large amount of fibrous feeds (ligno-
1. cellulosic materials) efficiently,

Ruminant can convert useless substances to useful products,

They have ability to utilize crop residues and other ago-
2. industrial byproducts,

They can detoxify many toxic ingredients present in feeds of

3. plant origin.

They can use non protein nitrogen sources like urea as a source
of nitrogen to meet part of their protein requirement.
4. They can also synthesis vitamins.
Why we need Rumen manipulation
Anaerobic fermentation of feeds in the rumen is beneficial for the host animal.

The co-existence of animal and its microbial eco-system

perform the fermentation process optimally.

Do we really need the manipulation of the rumen ecosystem?

Yes

High producing varieties of livestock have been

evolved world over by rumen manipulation

Improved feed Better feed Improved production

utilization conversion efficiency performance of the animals

Decreased methane
production
Major objectives of Rumen manipulation
1. Enhance fibro lytic activity:

2. Increase microbial protein synthesis:

3. Reduction in proteolysis

4. Reduction in methanogenesis

5. Prevention of acidosis

6. Shifting acetate to propionate production

7. Novel microbes: can be tailored to synthesis Amino acids

8. Metabolism of plant toxins

9. Synthesis of useful secondary metabolites

Methods of manipulation of rumen
microbes
1. Genetic 2. Non genetic

Probiotics
Antibiotics
Defaunation
Buffering agents
Organic acids
Probiotics
The term "Probiotic" which was a Greek word and meaning
for life was first of all used by the Parker (1974).
Defined probiotics as "A live microbial feed supplement
which beneficially affects the host animals by improving its
intestinal microbial balance." This definition encompasses
single strain or a mixture of two or more
FDA used the term direct fed microbes (DFM) instead of
probiotic.
The commonly used probiotics for animal feeding are
broadly divided into two categories i.e., bacterial origin and
yeast origin.
 Primary microorganism used as
probiotics
Bacterial origin
Bacillus licheniformis
Bacillus subtilis
Bifadobacterium adolescentis
Bifadobacterium animalis
Bifidobacterium bifidus
Bifadobacterium infantis
Bifadibacterium longum
Bifidobacterium peudolongum
Bifidobacterium suis
Bifadobacterium hermophilum
Streptococcus intermdius
Lactobacillus acidophilus
Lactobacillus bulgaricus
St reptococcus thermophilus
Lactobacillus casei
Lactobacillus cellobiosus
Lactobacillus delbrueckii
Lactobacillus fermentum
Lactobacillus lactis
Lactobacillus plantarum
Lactobacillus salivarius
Lactobacillus sporogens
Lactobacillus reuteri
Streptococcus intermdius
Yeast origin
Lactobacillus brevis Aspergilus
oryzae
Saccharomyces cerevisiae
Properties of micro-organism (Probiotic)

Resistance to low pH and bile salt,

A normal inhabitant of the gut in the target animal
species,
Able to survive, colonize and multiply at a faster rate in the
gut,
Viable product can be formed at industrial scale for its
commercialization,
Stable and viable during long storage and field conditions,
Antimicrobial property with other microbes,
Must produce beneficial effect in host animals.
Benefits of probiotics
Growth promotion,
Improved feed conversion efficiency,
Better absorption of nutrients by control of gut epithelial
cell proliferation and differentiation,
Improved metabolism of carbohydrate, calcium and
synthesis of vitamins,
Neutralization of anti nutritional factors i.e., trypsin
inhibitor, phytic acid etc,
Microbial enzyme production, compensating for deficient
intestinal enzyme activities of the host,
Elimination or control of intestinal microorganisms
producing sub clinical or clinical diseases,
Stimulation of non specific and specific immunity at the
intestinal level.
When probiotics are effective
After birth to encourage the
early establishment of beneficial
rumen microflora,
In the presence of enteric
pathogen such as E. coli,
Salmonella, Coccidia,
During environmental or
mangemental stress.
Before and after transportation,
At weaning,
Following over eating or
antibiotic
Ketosis
Bloat,
Effect of probiotics
Increase the total number of rumen
bacterial population along with higher
1. Rumen proportion of cellulolytic bacteria
microbial Observed higher number of amylolytic,
pectinolytic and xylanolytic bacteria in the
population rumen liquor of animals fed Lactobacillus
cellobiosus along with yeast.

Yeast have a buffering effect in the rumen

medium and prevent sharp drops in rumen
2. Rumen pH and thus stabilize the pH even in the
high concentrate fed animals.
pH The higher ruminal pH in yeast culture fed
animals was associated with a lower lactate
concentration
 3. Volatile Stimulation in VFA production as well as
proportion propionate production,
fatty acid Even we can increase in the proportion of
production acetate

4. Lower ruminal ammonia nitrogen

Ammonia concentration and increases microbial protein
synthesis.
nitrogen

5. Rumen Increases ruminal hydrolytic enzyme activity

enzyme like carboxymethyl cellulase, amylase, xylanase,
-glucosidase and protease
profile
Effect on animal performance

Feed intake, Glutamic acid is produced,

Nutrient digestibility,

Growth and feed conversion efficiency,

Milk production,

Incidence of diarrhea
DEFAUNATION
The process of making the rumen of animals free of rumen
protozoa is called defaunation and the animal is called
defaunated animal.
The role of rumen ciliate protozoa on the performance of
host animals became debatable issue when Becker and
Everett (1930) demonstrated that rumen protozoa were
non-essential for growth in lambs.
The recent reports reflect that though protozoa may be
non essential for ruminant, still they have significant role
to play in the rumen metabolism specially to stabilize the
rumen pH
Methods of defaunation
Isolation of new born animals
Preventing them from any contact with the adult
ruminant animals
Newborn should be separated 2 to 3 days after birth

Dietary manipulation
The ciliate protozoa are very much sensitive to
change in rumen pH (below 5)
Offering high energy feed (esp. cereals to the
starved animals for 24hours

Chemical treatment
Copper sulphate, Manoxol and Sodium lauryl
sulphate
Effects of defaunation
Bacterial population increased
Rumen Increases amylolytic , decreases cellulotytic
microbes bacteria
Fungal population in the rumen also increase

Rumen protozoa engulf the readily fermentable

carbohydrate (starch) which is stored as
amylopectin
Rumen pH Decrease the rate of carbohydrate fermentation
resulting in a lower pH in the rumen of
defaunated animals

About 40-60% of hydrolytic enzyme activity is

VFA found in the rumen protozoa
Production Increases propionate: ciliate protozoa engulfed the
feed particle and degrade it to acetate and butyrate
 In defaunated animals, the number as well as
Ammonia N activity of rumen bacteria increase resulting in
concentration more uptake of ammonia by bacteria and
ruminal ammonia concentration is reduced

Microbial The efficiency of rumen bacterial growth is

protein enhanced and more microbial protein flows
synthesis from reticulo-rumen to duodenum

Decreases carboxymethyl cellulase enzyme

Enzyme profile activity
Higher ruminal protease enzyme activity

Considerable decrease the methane production

Rumen protozoa contribute hydrogen moiety for
Methane the production of methane by the methanogenic
production bacteria
Symbiotic attachment methanogens have with
ciliate protozoa
Effect on animal performance
Defaunated animal losses appetite for few day after defaunation and
then stabilizes
The reduced digestibility of fiber fraction in defaunated animals
Defaunated animals are more susceptible to the bloat than normal
animals
Defaunated animals are more sensitive to copper toxicity.
Bird and Leng (1984) observed increased wool growth in the
defaunated lambs.
The availability of metabolizable energy was higher in defaunated
animals due to reduction of energy loss in methane production
Total heat production of animal was also significantly lower in absence
of rumen ciliate protozoa.
Osman et al. (1970) and Ramprasad(1981) observed reduced feed
conversion efficiency in the absence of ciliate protozoa
Ionophores
Ionophores are antimicrobial compounds that are commonly fed
to ruminant animals to improve feed efficiency.
These antimicrobials specifically target the ruminal bacterial
population and alter the microbial ecology of the intestinal
microbial
Ionophores transport ions across cell membranes of susceptible
bacteria, dissipating ion gradients and uncoupling energy
expenditures from growth, killing these bacteria.
Lasalocid (BovatecTM), Monensin (RumensinTM) and
Laidlomycin propionate (CattlystTM) are ionophores with
combined yearly sales of more than $150 million.
Benefits of Ionophores
Monensin has an inhibitory effect on methane and stimulatory
effect on propionate production,
Ionophores causes reduction of dietary protein deamination,
resulting in less ammonia urinary excretion, and a decrease in
lactic acid production which results in a reduction in ruminal
acidosis and liver abscesses,
The increases in energy availability and efficiency of feed
utilization by the ruminant animal,
Ruminal methane production is decreased 30% by monensin
treatment
Ionophores increases gross energy (GE) available to the animal
by 5.6%
Ionophores reduces morbidity and mortality among feedlot
animals by reducing the incidence of acute and sub-acute
ruminal acidosis, bloat, and bovine emphysema.
Mechanism of action
Because ionophores are highly lipophilic, they rapidly dissolve into
bacterial cell membranes
Ruminal bacteria maintain high intracellular potassium and low
intracellular sodium concentrations and conversely, the ruminal
environment contains high sodium and low potassium concentrations.
Monensin, is a metal/proton antiporter that can exchange H+ for either
Na+ or K+ Once inserted in the membrane,
Monensin exchanges intracellular potassium ions for extracellular
protons, or extracellular sodium for intracellular protons . Because the
potassium gradient is greater than the sodium gradient, protons
accumulate inside the bacterium.
The bacterium reacts to this cytoplasmic acidification by activating a
reversible ATPase to pump these protons out of the cell. Additionally,
other ATP-utilizing primary pumps for Na+ removal and K+ uptake are
activated to reestablish ion gradients.
Resulting in the uncoupling of ATP hydrolysis from growth, thereby
decreasing intracellular ATP pools, leading to cellular death
Effect of Ionophore antibiotics
Ionophores are potent antimicrobial compounds that improve
production efficiency and health in cattle by altering the
composition of the ruminal microbial ecosystem,
Ionophores reducing the incidence of illnesses related to the
ruminal fermentation (e.g., bloat, bovine emphysema).
However, like all other antimicrobial compounds, concerns have
been raised about the development of antimicrobial resistance
and the potential for the transfer of cross resistance to antibiotics
used in human medicine.
It appears that ionophores do not promote the development or
dissemination of antibiotic resistance, likely due to the complex
nature and high degree of specificity of ionophore resistance
mechanisms.
Therefore, it appears that the use of ionophores will continue to
be an important tool in improving animal production efficiency.
Non ionophore antibiotics
Non ionophore antibiotics are added in the sub-
therapeutic concentration for two principle reasons:
1. To act prophylactically against specific
organisms or group of organisms.
2. To increase the rate of weight gain, and
thereby improve feed conversion or efficiency
Mode of action
The antibiotics directly influence the
Metabolic effect rate or pattern of metabolic processes
of the animal

The antibiotics alter bacterial

Nutrient sparing
population, resulting in a
effect
conservation of nutrient

The antibiotics suppress bacteria

Control of
causing clinical or subclinical
subclinical disease
infections

Modification of The antibiotics alter the rumen

ruminal microbial population to improve
fermentation fermentation efficiency
Non ionophore antibiotics used in ruminants
Antibiotic Producing Chemistry Spectrum MOA
org. INHIBITION
Avoparcin Streptomuces Glucopeptide Narrow- G+ C W synthesis
candidus
Bacitracin Bacillus Polypeptide Narrow-G+ C W synthesis
subtilis
Chlorotetracy- S.aureofacien Teracycline Broad Protein synth.
cline s
Flavomycin S.bamerginsis P glycopeptide Narrow- G+ C W synthesis

Neomycin S.fradiae Aminoglycosi Narrow- G+ Protein synth.

de
Oxytetracycline S.rimosus Tetracycline Broad Protein synth.

Spiramycin S.ambofacien Macrolide Narrow- G+ Protein synth.

s
Tylosine S.fradiae Macrolide Narrow- G+ Protein synth.

Virginiamycin S.virginae Macrocyclic Narrow- G+ Protein synth.

lactone
Buffers
Added to the diet to aid in resisting pH changes of the rumen
when acids are present
Large quantities of organic acids produced in rumen by
microbial fermentation
Reduces incidence of acidosis on high grain diets
SALIVA is principle source of rumen-buffering agents

Performance is variable
-2 - 5% improvement in ADG & FE
Sources of buffers
Sources:
Sodium Bicarbonate: 0.75 - 1.5% diet DM
Limestone: 1.0% diet DM
Sodium Bentonite: 1 - 2% diet DM
Magnesium oxide: 0.5 - 0.75% diet DM
Manipulation of ruminal fermentation
by plant extracts
Plant extracts have been used for centuries for various
purposes as traditional medicine and food preservatives,
The antimicrobial activity of plant extracts - secondary
plant metabolites (saponins, tannins, and essential oils)
and selective inhibitors of methanogens
Plants exhibiting anti-methanogenic activity include:
Equisetum arvense, Lotus corniculatus, Rheum palmatum,
Salvia officinalis, Sapindus saponaria, Uncaria gambir and
Yucca schidigera
Manipulation rumen microbes with
organic acids
Manipulating rumen fermentation of treatment of
roughages and concentrates with organic acids could
improve rumen efficiency by
Maintaining higher pH,
Optimum utilization of ammonia nitrogen,
Reducing methane production,
Increasing microbial protein synthesis and
Increases essential VFA synthesis.

Organic acid treatment with sodium di malate or urea

Urea treated wheat straw
AREA OF FUTURE RESEARCH
Screening of non conventional animal feeds specially tree leaves for
anti-protozoal activity.
Standardization of defaunation method for its implication at farmer's
level.
New species/ strains of microorganism should be screened to use as
probiotic.
Mechanism of action of probiotic should be studied thoroughly.
Reduction in methanogenesis to improve availability of digestible
energy and reduce environmental pollution.
Production of suitable strain of recombinant microorganisms and their
propagation in the rumen for efficient detoxification of plant toxins,
reduction in methanogenesis, higher cellulolysis, reduced ruminal
proteolysis (deamination).
 Conclusion
Rumen is an natural fermentative anaerobic system which should be
manipulated essentially by altering the composition of rumen
microflora.
There is ample scope to manipulate the rumen by feeding local plants
or tree leaves or agro industrial by products to defaunate the animals
for improving its productivity.
Introduction of naturally occurring microorganism from digestive
system of one species to another species for efficient degradation of
plant toxins as well as for efficient utilization of nutrients will be one of
the major thrust area in near future for rumen manipulation.
Manipulation of rumen microorganism for efficient ruminal
fermentative digestion has an enormous potential to increase animal
productivity .
However in tropical countries, more emphasis should be given for
manipulating the rumen to increase cellulolytic activity for efficient
utilization of low grade roughage.
References
Rumen Manipulation to Improve Animal Productivity by
A. Santra and S. A. Karim.
Ionophores: Their Use as Ruminant Growth Promotants
and Impact on Food Safety by T. R. Callaway and T. S.
Edrington.
Opportunities to improve fiber degradation in the rumen
by Denis O. Krause and Stuart E. Denman.
Rumen microbial ecosytem by P.N.Hobson and
C.S.Stewart.
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