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James E. Nocek, Ph.D.
Spruce Haven Farm and Research Center
Auburn, NY
2,000 Cow Research Dairy
What Is The Difference?
Yeast Culture Component
LIVE-CELL YEAST
BREWERS YEAST
YEAST CULTURE
Budding yeast cells
Metabolites
During fermentation, yeast cells release amino acids, organic
acids, minerals, vitamins and enzymes into the environment,
which stimulates the growth of certain rumen microbes.
Select Strain
Saccharomyces
cerevisiae
Select Strain
Saccharomyces
cerevisiae
Stage 2
Stage 1
Corn
Germ
Meal
All yeast cultures are
manufactured using the
same basic process
used at Vi-COR
Corn
Germ
Meal
QA
Large Scale
Prepared
substrate for
initial
fermentation
& yeast
growth
Anaerobic
Fermentation
KEY DIFFERENCES IN PROCESS
Strain Selection
Scientifically
formulated die
designed to
maximize
fermentation
metabolites
Diet Fed To Organism
QA
Carrier Selection
Low Temperature Drum Dryer
Low Temperature
QA
QA
Cooling Phase
Drying
Low Temperature
Drying
Product Uniformity & Consistency
Secondary
Nutrient Blender
Finished Product
QA
Cold Air Induction
Evaporation System
Finished Product
Distributed for Packaging
Cold Air Induction
Evaporation System
Distributed for Packaging
QA
1
Efficacy Studies with A-Max®
Yeast Culture
RESEARCH SUMMARY
In House Study For Initial Formulation Test
Test
Product
RB 40
In house
Test
Researcher
A-Max® vs. DV
Ag Tech
Product
Researcher
Results
Higher Lactic Acid Utilizing Bacteria,
Higher Gas Production
Results
RB 7
Artificial Rumen
A-Max® vs. DV
Hoover
Higher Microbial Protein Microbes Contained Less Ash
Higher Rumen pH
J.Dairy Sci. (2002) 85:2009
RB 5
Artificial Rumen
A-Max® Conc. vs. A-Max® XTRA
Hoover
No Difference Between Conc. & XTRA
No Difference in Rumen Parameters
RB 4
Artificial Rumen
A-Max® XTRA vs. A-Max® ULTRA
Hoover
RB 2
Artificial Rumen
A-Max® ULTRA vs. Live Cell
Hoover
Increased NDF Digestibility
Artificial Rumen
A-Max® XTRA vs. A-Max® Liquid
Hoover
Dry Matter Digestibility Was Higher for Both Yeast
Products Over Control
RB 1
Artificial Rumen
A-Max® Liquid vs. A-Max® XTRA
Hoover
Liquid Enhanced Rumen Function Comparable to XTRA
RB 10
Artificial Rumen
A-Max®
Hoover
Enhanced Rumen Function
RB 9
RB 12
Artificial Rumen
A-Max® Conc.
Hoover
Dose Response. 2-4 oz/h/d Was Optimal To Enhance
Rumen Microbial Metabolism
RB 26
Artificial Rumen
A-Max® ULTRA vs. YeaSacc
Hoover
ULTRA and YeaSacc Equally Stimulated Rumen
Microbial Metabolism
• West Virginia University
• Spruce Haven Farm and Research Ctr
• University of California, Davis
YEAST CULTURE
METABOLITES ? ? ? ? ? ?
WHAT ARE THEY ? ? ? ?
The Next
Evolution In
Yeast Culture
WHAT DO THEY DO ? ? ? ?
The Yeast Cell
313
3345
METABOLITE ANALYSIS
University of Texas -Cancer Research Center, 2004
2
MECHANICAL FRACTURE OF CELL WALL
BALL
MILL
SHEAR
PUMP
PROBLEM: Cross-sections cell wall exposing
less surface area of layers.
HOW ENZYMES WORK
THIS REACTION HAPPENS MILLIONS / SECOND
313
3345
MOS
METABOLITE ANALYSIS
PROTEIN
University of Texas -Cancer Research Center, 2004
CARBOHYDRATE COMPOSITION OF MOS PRODUCTS
CELMANAX® FINGERPRINT
CARBOHYDRATE CHEMISTRY
Monosaccharide analysis
d-fucose
3.1%
d-galactosamine
27.9%
(as n-acetyl d-galactosamine)
d-glucosamine
7.0%
(as n-acetyl d-glucosamine)
d-galactose
3.4%
d-glucose
49.3%
d-mannose
9.3%
Hydrolyzed assay
Free assay
Hydrolyzed assay
Free assay
LEADING COMPETITOR
0%
19.6%
0%
5.2%
0%
3.7%
8.5%
0%
0%
44.6%
45.0%
27.3%
16.8%
78.0%
0%
1.0%
63.0%
32.3%
FUCOSE
80
GLUCOSAMINE
MANNOSE
60
% 40
20
GALACTOSAMINE
GALACTOSE
GLUCOSE
0
GALACTOSE
MANNOSE
1 2 3
4 5
FUCOSE
6 7
8 9 SUGAR
SAMPLE
S
In the laboratory, d-mannose is measured to determine
relative differences in MOS content among products.
3
PROCESS
BIOCHEMICAL ENZYME REACTION
YEAST CULTURE HYDROLYZED YEAST
+
+
Yeast Cell Metabolites
Fermentation
Metabolites
SOURCE OF YEAST CELL WALL
WARM WATER
pH CONTROL
ENZYMES
TIME
MECHANICAL ACTION
BLENDER
APPLY TO CORN GERM
CARRIER
DRY
MOS IS A FOOD SOURCE FOR GOOD BACTERIA
MOS – MANNAN OLIGOSACCHARIDE
A FERMENTABLE CARBOHYDRATE FOR
LACTOBACILLUS AND BIFIDOBACTERIUM
IN THE LOWER DIGESTIVE TRACT
d - mannose
Tips on pili look for chemistry
on epithelial tissue in gut
d-mannose bonded
to bacteria pili
HYDROGEN
BONDS
d-mannose has a relationship
to bacteria pili
Interferes with recognition
between pathogens and
epithelial tissue
4
SALMONELLA DUBLIN
Ω
ΩΩΩΩ
ΩΩ
Ω
Ω
ENTEROBACTERIACEA
E. coli Adhesin
Bacterial adherence normally critical for
successful colonization, Adhesins attach
to host tissue. Mannose will bind to
adhesins and interfere with colonization.
E. coli Type 1 Fimbriae (pili)
<
< <<<
<
Pili will bind by recognizing
oligomannose-like receptors of host
tissue.
Type 1 fimbriae are mannose sensitive,
binding is inhibited by mannose.
ENTEROTOXINS CAUSE INFLAMATION
CELMANAX® INHIBITS
ATTACHMENT TO
TISSUE
Type 1 pilus-mediated bacterial adherence to the mouse bladder epithelium
SHIGELLOSIS
PNEUMONIAE
Opportunistic
BUBONIC PLAGUE
bacterium being enveloped by
bladder epithelial cells
Agglutination of E. coli with Celmanax
bacterium making intimate contact
Type 1 pili can be seen with radiating
tips, these pili mediate contact with
epithelial cell membrane
®
Celmanax® mediated agglutination of E. coli F18
1.30E+09
1.10E+09
cfu/mL
9.00E+08
7.00E+08
5.00E+08
3.00E+08
1.00E+08
This video demonstrates how E. coli is agglutinated to the MOS
component of Celmanax® when the liquid is introduced to a media
contaminated with E. Coli. Total agglutination time 00:30 seconds
1
control
2
20mg/mL
3
40mg/mL
5
CARBOHYDRATE COMPOSITION OF MOS PRODUCTS
Monosaccharide analysis
Free assay
Hydrolyzed assay
d-fucose
3.1%
d-galactosamine
27.9%
(as n-acetyl d-galactosamine)
d-glucosamine
7.0%
(as n-acetyl d-glucosamine)
d-galactose
3.4%
d-glucose
49.3%
d-mannose
9.3%
Hydrolyzed assay
Free assay
LEADING COMPETITOR
0%
19.6%
0%
5.2%
0%
3.7%
8.5%
0%
0%
44.6%
45.0%
27.3%
16.8%
78.0%
0%
1.0%
63.0%
32.3%
In the laboratory, d-mannose is measured to determine
relative differences in MOS content among products.
MYCOTOXIN CAPABILITY
Celmanax pH 3
Celmanax® VS. MTB 100
Celmanax pH 6.5
MTB 100 pH 3
100
MTB 100 pH 6.5
% Binding
90
80
70
60
Romer Labs
50
40
University of
Missouri
30
20
10
0
AFLATOX
YEAST GLUCAN
AFFINITY
& BINDING
T-2
ZEARLE
DON
FUMONIS
OCHRA
TOXIN
How C. parvum Works
The two most critical factors in
preventing C. parvum outbreaks
Nydam and Mohammed 2005
• Cannot grow or multiply in the
environment
• Needs host for oocytes to proliferate
• Hygiene of calf rearing area
• Prevent entry of newly infected calves in
high density areas
6
Maternity Pen
~8.7% preweaned
mortality
~62% of preweaned
mortality due to scours
56% of heifers removed at
birth
~30% nurse the dam
~40% FPT
Salmonella/E. coli
“Crypto”
Rota/Corona
Coccidia
NAHMS 2002
How does C. parvum survive in the
host animal? (cont)
• invagination of the host cell plasma
membrane, which extends along the
surface of the sporozoite and eventually
completely surrounds it,
• “Occupation": formation of a
“parasitophorus” vacuole where the
parasite undergoes further development in
a unique intracellular but extra-cytoplasmic
location.
How does C. parvum survive in the
host animal?
Crucial steps in the pathogenesis of
cryptosporidium
• Ingestion of oocyte
• Attachment to intestinal epithelial cells
• Invasion of host intestinal epithelial cells
(Sporozoites attach to host cells by their
anterior pole).
Crypto Infestation Process
OOcytes
Fecal Matter
Ingestion
Feed
Attachment
Invasion
Invagination
Occupation
Preventative Dietary Therapy
• If attachment is critical to C parvum
survival:
• Then:
– Prevent attachment!
• Find something else that the bacteria like
to attach to that is:
– not part of the anatomy,
– inactivates the pathogen
– can be removed
Effect of Celmanax® on C. parvum infectivity in
bovine and porcine intestinal epithelial cells in-vitro
Rural Technologies, Brookings SD
• Sporozoites of C. parvum were pretreated with
Celmanax®, BSM or buffer and then allowed to infect
cells
• First slide shows the degree of attachment of treated
sporozoites to the intestinal cells
• Second slide shows the degree of infectivity of
intestinal cells by treated sporozoites
7
Celmanax® Liquid - In vitro inhibition Of Cryptosporidium
parvum in bovine intestinal epithelial cell line
Effect of Celmanax® on Cryptosporidium
infectivity rate in porcine intestinal cells
Celmanax®
0.3
0.25
0.2
0.15
0.1
0.05
0
Control
BSM
ViCor 40
mg/mL
ViCor 20
mg/mL
ViCor 2
mg/mL
Treatment
Trial Design
THE EFFECT OF CELMANAX® LIQUID ON CRYPTO OOCYTE
SHEDDING, FECAL AND DEHYDRATION SCORE:
A FIELD DEMONSTRATION
J. E. Nocek, Spruce Haven Farm and Res Ctr, Auburn, NY
• 16 calves 10-12 days old randomly assigned to
treatments
• Treatments –
Control pasteurized whole milk
Pasteurized whole milk with Celmanax®
8mL/calf/day
• Treatments fed for 5 days
• Fecal samples were taken at the start and finish of the
trial and evaluated for # of oocytes, fecal score and
dehydration score
Effect of Celmanax® on crypto infected calves
Effect of Celmanax® on crypto oocyte shedding in sick calves
5
Control d10
100000
Control d15
Celmanax® d10
Celmanax® d15
10000
4
score
No. of oocytes
1000000
a
a
3
b
a
2
b
Control d10
Control d15
Celmanax® d10
p=0.03
Celmanax® d15
1
fecal score
dehydration score
8
Conclusions
Celmanax® Liquid
On Farm Trial
• Calves supplemented with Celmanax® Liquid
demonstrated 3 fold less oocyte shedding within
5 days after supplementation.
• Fifteen day fecal and dehydration scores were
significantly less for calves supplemented with
Celmanax® Liquid.
Brian and Christy Sherber
Cold Spring Co-op, Cold Spring, Minnesota
Prepared by:
Ashley Kohls
Calf and Heifer Specialist
Land O’ Lakes Purina Feed, LLC.
And Peter Sook
Mortality at the end of the trial
Trial Design
No. of dead calves
5
• 17 calves on Celmanax® Liquid and 22 calves
on control treatment. Calves on Celmanax®
were lighter at the start of the trial compared to
control calves.
• Calves were fed 22-20 CMR plus free (amplifier
max) 18% starter and free choice water.
• Calves had tested positive for crypto at the start
of the trial. But no crypto related data was
collected.
• Mortality, and wt gain was recorded every week
and ADG at the end of the trial.
4
3
2
1
0
control
Celmanax
Treatments
Testimonial D-2
General Observations
Testimonial Report on Feeding Celmanax® at the School of Veterinary Medicine Dairy,
University of Wisconsin-Madison
• Calves on the Celmanax® treatment
weaned easier with less stress to the calf.
• Mortality was reduced in the Celmanax®
treatment.
• No significant wt. difference was observed
between treatments
Dr. Sheila McGuirk, DVM,Ph.D and Dr.
Keith Poulson, DVM
From down cows, cows
recovering from surgery
or cows with
Salmonellosis, they
have developed a
confidence in the
product as part of the
treatment to get cows
back to full feed and
normal rumen activity.
9
Liquid
Liquid
32 oz. dose bottle
2.5 gal. jug
275 gal. tote
Calves = 4
Calves = 43
55 gal. drum
Calves = 4760
For Use In Milk & Milk Replacer
Calves = 951
Currently Shipping tanker loads
to Idaho for further processing
or as a Drench
10