Immune Function - Immune Function - The New Frontier in The New Frontier in Animal ProductionAnimal Production

Steve Puntenney, Ph.D.Steve Puntenney, Ph.D.

OmniGen Research, LLCOmniGen Research, LLC

Mission StatementMission Statement

OmniGen Research is dedicated to OmniGen Research is dedicated to expanding the boundaries of science to expanding the boundaries of science to benefit animal health and productivity benefit animal health and productivity through the development of natural through the development of natural products for the livestock industryproducts for the livestock industry..

History of OmniGen ResearchHistory of OmniGen Research

1999 1999 HBS identified as a significant issue in HBS identified as a significant issue in OR & WA DairiesOR & WA Dairies

May 2001 Meetings w/ consulting veterinarians May 2001 Meetings w/ consulting veterinarians in WA indicated a high failure rate of in WA indicated a high failure rate of

autogenous vaccines & need for other autogenous vaccines & need for other intervention identifiedintervention identified

Feb 2002 Prototype OmniGen placed on 6 OR & Feb 2002 Prototype OmniGen placed on 6 OR & WA DairiesWA Dairies

May 2002 OmniGen Research, LLC formed May 2002 OmniGen Research, LLC formed with NEF to discover MOAwith NEF to discover MOA

Mar 2003 PAP began distribution of OAFMar 2003 PAP began distribution of OAF Jan 2004 OmniGen-AF Licensed to PAPJan 2004 OmniGen-AF Licensed to PAP

History of OmniGen ResearchHistory of OmniGen Research

July 2005July 2005 Construction on the OmniGen Construction on the OmniGen Research LabResearch Lab

June 2006June 2006 Established OmniGen Established OmniGen Research Research Farm Farm

Feb 2007Feb 2007 Beginning of construction of dairy Beginning of construction of dairy and lab animal facilities and lab animal facilities

Overview of presentationOverview of presentation

Part 1Part 1 Acquired and innate immunityAcquired and innate immunity

• MacrophagesMacrophages• NeutrophilsNeutrophils

Nutrition and immunity during Nutrition and immunity during transitiontransition

Immune function and udder Immune function and udder healthhealth

Part 2Part 2 OmniGen Mastitis StudiesOmniGen Mastitis Studies Mycotic AbortionsMycotic Abortions Mycotoxin BindingMycotoxin Binding OmniGen Use RatesOmniGen Use Rates

Adaptive (antibody-mediated) immunityAdaptive (antibody-mediated) immunity

Janeway et al., 2005

Vulnerable

Impact of periparturient Impact of periparturient immunosuppressionimmunosuppression

Increased susceptibility to diseasesIncreased susceptibility to diseases

-mastitis-mastitis

-metritis/retained placenta-metritis/retained placenta

-foot infections-foot infections

-respiratory infections-respiratory infections

-gastrointestinal disorders-gastrointestinal disorders

-lowered peak milk production-lowered peak milk production

Why focus on immune function?Why focus on immune function?

“ “A .5 kg increase in peak milk production is A .5 kg increase in peak milk production is equal to 108 kg of additional milk per equal to 108 kg of additional milk per lactation” (Utah State University)lactation” (Utah State University)

Innate immunity Innate immunity Allows time for adaptive immunity to developAllows time for adaptive immunity to develop

First line of defenseFirst line of defense Evolutionarily ancientEvolutionarily ancient Several componentsSeveral components

– Epithelial barriersEpithelial barriers– HClHCl– Digestive enzymesDigestive enzymes– Cellular componentCellular component

MacrophagesMacrophages NeutrophilsNeutrophils Natural killer (NK cells)Natural killer (NK cells)

Opsonization is the coating of microbes with Opsonization is the coating of microbes with complement components, such as C3b complement components, such as C3b (also IGg & antibodies, but later).(also IGg & antibodies, but later).

Opsonized particles are more easily Opsonized particles are more easily

phagocytosed due to the presence of phagocytosed due to the presence of complement receptors on the plasma complement receptors on the plasma membrane of phagocytic cellsmembrane of phagocytic cells

Picture of macrophage surface using scanning electron microscopy

Opsonization of Bacteria by C3b

C3bC3b

Cytokine releaseCytokine release

Neutrophil TrafficingNeutrophil Trafficing

Burton and Erskine, 2003

L-selectin

IL-8

Phagocytosis by the neutrophilPhagocytosis by the neutrophil

Bacterium

Recognition

Killing mechanisms: Respiratory burst Killing mechanisms: Respiratory burst via via ROSROS

Phagocytosis NADP oxidase

superoxide anion (O.

2-)

H2O2

O2+

SuperoxidedismutaseBacteriocidal

hypochloriteion OCl- Neutrophil

myeloperoxidase

Other oxygen-derivedfree radicals (OH.)

Macrophages: myeloperoxidase-Independent mechanism

killing

killing

ROS: Good and BadROS: Good and Bad

Good: kill pathogensGood: kill pathogens

Bad: tissue damageBad: tissue damage

““Immune cells” have Immune cells” have high intrinsic ROS high intrinsic ROS generation and, generation and, therefore, have high therefore, have high anti-oxidant demandanti-oxidant demand

““Immune cells” have Immune cells” have membranes high in membranes high in PUFAs which are PUFAs which are susceptible to ROS-susceptible to ROS-mediated damagemediated damage

Chew and Park, 2002

A primary mechanism by which nutrients support A primary mechanism by which nutrients support immunity is via protection against ROSimmunity is via protection against ROS

Phagocytosis NADP oxidase

superoxide anion (O.2-)

H2O2

O2+

SuperoxidedismutaseBacteriocidal

hypochloriteion OCl- Neutrophil

myeloperoxidase

Other oxygen-derivedfree radicals (OH.)

Macrophages: myeloperoxidase-independent mechanism

Cu/Zn

Anti-oxidants (scavenge free radicals)vitamin E, vitamin C

zinc, selenium, carotenoids, flavenoids

Nutrition and immunology at Nutrition and immunology at parturitionparturition

Three factors conspire to affect nutrition and Three factors conspire to affect nutrition and immunity at parturitionimmunity at parturition

– Surge in cortisolSurge in cortisol– Decline in feed intakeDecline in feed intake

NEFA spikeNEFA spike Ketone body spikeKetone body spike

– Impaired calcium homeostasisImpaired calcium homeostasis

Impact of periparturient Impact of periparturient immunosuppressionimmunosuppression

Increased susceptibility to diseaseIncreased susceptibility to disease

-mastitis-mastitis

-metritis/retained placenta-metritis/retained placenta

-foot infections-foot infections

-respiratory infections-respiratory infections

-gastrointestinal disorders-gastrointestinal disorders

Periparturient immunosuppressionPeriparturient immunosuppression

ParturitionParturition Drying offDrying off ShippingShipping Over-crowdingOver-crowding Environmental Environmental

(heat/hold)(heat/hold) Excess handlingExcess handling

Neu

trop

hil L

-sel

ectin L-selectin

Source: Burton and Erskine, 2003

Cortisol brings about immunosuppressionCortisol brings about immunosuppression

Healthy mammary tissue Mammary tissue in stress

Source: Burton and Erskine, 2003

Cytokines released from macrophages Cytokines released from macrophages signal neutrophil migrationsignal neutrophil migration

Increased mastitis incidence due to Increased mastitis incidence due to periparturient immunosuppressionperiparturient immunosuppression

Barkema et al., 1998

PrimiparousMultiparous

Dry Matter Intake during TransitionDry Matter Intake during Transition

6

8

10

12

14

16

18

20

22

Day Relative to Parturition

DM

I, kg

.

Control

Anionic

Puntenney & French 2006Puntenney & French 2006

Energy balance and parturitionEnergy balance and parturition

Puntenney & French 2006

-15

-12

-9

-6

-3

0

0 1 2 3

Week Relative to Parturition

EN

E B

alan

ce (

MC

al/K

g)

Control

Anionic

Plasma NEFAPlasma NEFA

Depressed dry matter intake and elevated blood NEFA Depressed dry matter intake and elevated blood NEFA concentrations during the 2 to 3 weeks prior to calving concentrations during the 2 to 3 weeks prior to calving have been associated with an increased occurrence of have been associated with an increased occurrence of ketosis (Duncan 1998, Goff 2006)ketosis (Duncan 1998, Goff 2006)

Mobilization of adipose tissue is related to the sudden Mobilization of adipose tissue is related to the sudden increase in demand for energy brought on by the onset increase in demand for energy brought on by the onset of lactation, not parturition itself (Goff 2002)of lactation, not parturition itself (Goff 2002)

Elevated NEFA is related to lower DMI at 14 d Elevated NEFA is related to lower DMI at 14 d postpartum and an increased incidence of metritis and postpartum and an increased incidence of metritis and mastitis (Goff 2006)mastitis (Goff 2006)

Impact of Plasma NEFA Concentration Impact of Plasma NEFA Concentration on neutrophil function (dairy)on neutrophil function (dairy)

Scalia et al. 2006

0

50

100

150

200

250

0 mM NEFA 2 mM NEFA

Oxi

dativ

e bu

rst

NEFAs inhibit ROSNEFAs inhibit ROSPhagocytosis NADP oxidase

superoxide anion (O.2-)

H2O2

O2+

SuperoxidedismutaseBacteriocidal

hypochloriteion OCl- Neutrophil

myeloperoxidase

Other oxygen-derivedfree radicals (OH.)

Macrophages: myeloperoxidase-independent mechanism

Cu/Zn

XX

ββ-hydroxybutyrate-hydroxybutyrate An indicator of KetosisAn indicator of Ketosis

Associated with negative energy balanceAssociated with negative energy balance

Cows with plasma BHB levels above 12 mg/dL, are Cows with plasma BHB levels above 12 mg/dL, are considered to be subclinically ketotic, above 15 mg/dL considered to be subclinically ketotic, above 15 mg/dL clinically ketotic (LeBlanc 2005)clinically ketotic (LeBlanc 2005)

Elevated BHB 14 d postcalving is associated with an Elevated BHB 14 d postcalving is associated with an increased incidence of metritis & mastitis (Goff 2006)increased incidence of metritis & mastitis (Goff 2006)

ββHB by Day Relative to ParturitionHB by Day Relative to Parturition

4

5

6

7

8

9

10

11

12

-1 0 1 7 14 21

Day Relative to Parturition

BH

B,

um

ol/

L

Puntenney & French 2006Puntenney & French 2006

Ketones suppress leukocyte functionKetones suppress leukocyte function

Hoeben Hoeben et alet al., 1999., 1999

HH22 OO

22 P

rodu

ctio

n

Pro

duct

ion

Ketones inhibit ROSKetones inhibit ROSPhagocytosis NADP oxidase

superoxide anion (O.2-)

H2O2

O2+

SuperoxidedismutaseBacteriocidal

hypochloriteion OCl- Neutrophil

myeloperoxidase

Other oxygen-derivedfree radicals (OH.)

Macrophages: myeloperoxidase-independent mechanism

Cu/Zn

XX

Hypocalcemia impairs immune cell Hypocalcemia impairs immune cell functionfunction

Kimura et al., 2006

Control cows

Cows with milk fever

Cal

cium

sig

nalin

g in

PB

MN

cel

ls

6

8

10

12

14

16

18

20

22

Day Relative to Parturition

DM

I, k

g.

Control

Anionic

What are the effects of OmniGen-AF What are the effects of OmniGen-AF on neutrophil function?on neutrophil function?

Respiratory burst Respiratory burst (ROS generation)(ROS generation)

Neutrophil NET Neutrophil NET formationformation

Phagocytosis AssayPhagocytosis Assay

Respiratory burst assayRespiratory burst assay

Combine -neutrophils (+/- OmniGen-AF)

-Cytochrome C -Zymosan A (from yeast)

Incubate 30 min, 37oC

Assess Cytochrome-C reduction by color change(655 nm)

Effects on Respiratory BurstEffects on Respiratory Burst

0

1

2

3

4

5

6

Control OmniGen

0 nM PMA

5 nM PMA

RO

S g

ener

atio

n

Omnigen-AF effect (P<0.01)

OmniGen-AF restored phagocytosis and OmniGen-AF restored phagocytosis and ROS generation in Stress ChallengeROS generation in Stress Challenge

Phagocytosis NADP oxidase

superoxide anion (O.2-)

H2O2

O2+

SuperoxidedismutaseBacteriocidal

hypochloriteion OCl- Neutrophil

myeloperoxidase

Other oxygen-derivedfree radicals (OH.)

Macrophages: myeloperoxidase-independent mechanism

Cu/Zn

Inactive neutrophils Activated neutrophils with NETs

Source: Brinkmann et al., 2004, Science

+ interleukin-8 (IL-8)

Production of NETs by neutrophils

Association of neutrophil “NETs” Association of neutrophil “NETs” with bacteriawith bacteria

S. aureus Salmonella Shigella

Gram + Gram - Gram -

Neutrophil NET assayNeutrophil NET assayNeutrophils +/- OmniGen-AF

Add to microplate with varyingConcentration of PMA (activator)

Incubate 30 minutes at 37oC

Add Sytox green (extracellularDNA marker)

Read in fluorescence plate reader485 excitation, 525 emmision

0

0.2

0.4

0.6

0.8

1

1.2

0 nM 0.5 nM 5 nM 50 nM

Control

OmniGen

OmniGen-AF enhanced NET release in immunosuppressed sheep neutrophils

Phorbol ester (PMA) concentration

Cha

nge

in D

NA

flu

ores

cenc

e re

lativ

e to

0 n

M P

MA

con

trol

OmniGen-AF as main effect (P<0.01)

Phagocytosis AssayPhagocytosis AssaySheep E. coli Strain 487

Neutrophils

Combine in ratio of 30 E. coli:1 neutrophil

Incubate 37oC, 2 hr

(allow killing of E. coli)

Add MTT

(live E. coli metabolize to formazin, purple)

Incubate 4 hr, 37oC in CO2

Stop rx, incubate 4 hr

Read at 655nm on plate reader

(+/- OmniGen)

OmniGen-AF enhanced phagocytosis in OmniGen-AF enhanced phagocytosis in stress challenged sheep neutrophilsstress challenged sheep neutrophils

0

0.2

0.4

0.6

0.8

1

1.2

1.4

1.6

1.8

2

Control OmniGen-AF

Kill

ing

of E

. co

li

P<0.01

Moving beyond molecular biologyMoving beyond molecular biology

DNA Cell function Organ function Whole animal Populations

L-selectinIL8R, ICEIL4Rthymopoeitin

MastitisMastitis

Mastitis is defined as an Mastitis is defined as an infection of part of the infection of part of the mammary glandmammary gland

Cost – about an Cost – about an average of $200/cow average of $200/cow annually. . . . .$2 billion annually. . . . .$2 billion to the industryto the industry

““A disease of man A disease of man transmitted to the cow”transmitted to the cow”

Impossible to eradicate, Impossible to eradicate, but practical to control.but practical to control.

Effect on Milk Composition & QualityEffect on Milk Composition & Quality

Rapid increase in somatic cell count Rapid increase in somatic cell count (leukocytes)(leukocytes)

Milk volume, milk fat, SNF, Milk volume, milk fat, SNF, protein, lactose, casein, protein, lactose, casein, calcium & shelf life calcium & shelf life

Whey protein, albumin, Whey protein, albumin, sodium, chloride & pH sodium, chloride & pH (bad things)(bad things)

Economic Loss due to MastitisEconomic Loss due to Mastitis

Anatomy of the Anatomy of the Mammary GlandMammary Gland

All pathogens enter through All pathogens enter through the streak canal viathe streak canal via

-employee hands-employee hands-contaminated -contaminated

beddingbedding-equipment insults-equipment insults-cow to cow -cow to cow

transfertransferDeLaval, Inc. 2006DeLaval, Inc. 2006

Teat Sphincter Streak canalTeat Sphincter Streak canal

PathogensPathogens Contagious Contagious

- - Streptococcus agalactiae Streptococcus agalactiae (very short life o/s the udder; responds to dc (very short life o/s the udder; responds to dc treatment and teat dips) treatment and teat dips) gramgram + +

- Staphylococcus aureus - Staphylococcus aureus (resistant to antibiotics; chronic) (resistant to antibiotics; chronic) gramgram + (CPS)+ (CPS) EnvironmentalEnvironmental

- - Other Strep. spp.Other Strep. spp.- Dysgalactiae Dysgalactiae (chronic mastitis) (chronic mastitis) gramgram ++- Uberis (contaminated bedding) gramgram ++

- - ColiformsColiforms

- - E. coli E. coli (hot mastitis, quick acting, (hot mastitis, quick acting, high temp & shock) high temp & shock) gramgram ––

- - Klebsiella spp. Klebsiella spp. (high incidence in (high incidence in wood shavings) wood shavings) gramgram – (CPS)– (CPS)

-Enterobacter sp. -Enterobacter sp. gramgram – –

-Aerobacter aerogenes -Aerobacter aerogenes gramgram – –

CPS type cell walls supress complement C3bCPS type cell walls supress complement C3b activity & resist phagocytosisactivity & resist phagocytosis Staph. aureusStaph. aureus

Other PathogensOther Pathogens Coagulase negative staph (live on skin; don’t cause clinical mastitis Coagulase negative staph (live on skin; don’t cause clinical mastitis gram + gram +

(CPS)(CPS) Psuedomonas aeruginosa (lives in water; responds to Cl) Psuedomonas aeruginosa (lives in water; responds to Cl) gramgram – (CPS)– (CPS) Corynebacterium pyogenes (antibiotic resistant) Corynebacterium pyogenes (antibiotic resistant) gram + (CPS)gram + (CPS) Nocardia spp. (standing water; acts like coliform) Nocardia spp. (standing water; acts like coliform) gram +gram + Mycoplasma spp. (spreads easily, shuts down milk Mycoplasma spp. (spreads easily, shuts down milk gram –gram –

production in 24 hours, equipment related)production in 24 hours, equipment related)occasional; dirty mastitis tubesoccasional; dirty mastitis tubes

Proteus spp. – not commonProteus spp. – not common gram- (CPS) gram- (CPS) Serratia spp. – chronic mastitisSerratia spp. – chronic mastitis gram – (CPS) gram – (CPS)

YeastsYeasts FungiFungi usually not significantusually not significant

Part 2Part 2

Mastitis Field Studies With OmniGenMastitis Field Studies With OmniGen Mycotic AbortionsMycotic Abortions Mycotoxin StudiesMycotoxin Studies OmniGen Use RatesOmniGen Use Rates

Case Study No. 7 - TexasCase Study No. 7 - Texas

- 3200 cow dairy, open lot design, silage based TMR3200 cow dairy, open lot design, silage based TMR

- Initiated an off/on/off protocol:- Initiated an off/on/off protocol:

Off - Aug. 1, 2003 - Sept. 27, 2003 (45 d)Off - Aug. 1, 2003 - Sept. 27, 2003 (45 d)

On - Sept. 28, 2003 - Dec. 30, 2003 (93 d)On - Sept. 28, 2003 - Dec. 30, 2003 (93 d)

Off - Jan. 1, 2004 - Mar. 3, 2004 (63 d)Off - Jan. 1, 2004 - Mar. 3, 2004 (63 d)

Case Study No. 7 - TexasCase Study No. 7 - TexasSummary of Cull/Dead Records by Feeding PeriodSummary of Cull/Dead Records by Feeding Period

Off Off OnOn OffOff (45 days) (93 days) (63 days) (45 days) (93 days) (63 days)

Total: Culls 109 163 144Total: Culls 109 163 144

Dead 36 45 43Dead 36 45 43

Respiratory: Culls 10 6 6Respiratory: Culls 10 6 6 Dead 4 7 5Dead 4 7 5

Digestive: Culls 12 8 18Digestive: Culls 12 8 18 Dead 14 15 16Dead 14 15 16

Mastitis: Culls 33 8 28Mastitis: Culls 33 8 28 Dead 1 4 6Dead 1 4 6

Case Study No. 7 - TexasCase Study No. 7 - Texas

Summary of culls and deaths by feeding periodSummary of culls and deaths by feeding period

Item Item Off Off OnOn (108 days) (93 days)(108 days) (93 days) Total: Culls 253 163 Total: Culls 253 163

Deaths 79 45 Deaths 79 45 ---------------------------------------------------------------------------------------------------------------------------------------------------------- Respiratory: Culls 16 6Respiratory: Culls 16 6 Deaths 9 7Deaths 9 7 Digestive: Culls 30 8Digestive: Culls 30 8 Deaths 30 15Deaths 30 15 Mastitis: Culls 61 8Mastitis: Culls 61 8 Deaths 7 4 Deaths 7 4

Case Study #9:Effect of OmniGen-AF on a Case Study #9:Effect of OmniGen-AF on a High SCC herd (4000 cows)High SCC herd (4000 cows)

Avg. SCC

350000

400000

450000

500000

550000

600000

7/3

0/2

006

8/1

3/2

006

8/2

7/2

006

9/1

0/2

006

9/2

4/2

006

10/8

/2006

10/2

2/2

006

11/5

/2006

11/1

9/2

006

12/3

/2006

12/1

7/2

006

Avg. SCC

OmniGen Added

Dominant Pathogen: Strep uberis; Minor Pathogen: E. coliDominant Pathogen: Strep uberis; Minor Pathogen: E. coli

Conditions Don’t Always Go Our Conditions Don’t Always Go Our Way!!Way!!

Case Study #10: Effect of OmniGen-AF Case Study #10: Effect of OmniGen-AF on a High SCC herd (3000 cows)on a High SCC herd (3000 cows)

OmniGen AddedOmniGen Added

Dominant Pathogen: Strep uberisDominant Pathogen: Strep uberis

High Incidence of New InfectsHigh Incidence of New Infects

PSCC | PSCCPSCC | PSCC

<200 | >=200 All<200 | >=200 All SCC 280 | 315 595SCC 280 | 315 595 >=200 12% | 13% 25%>=200 12% | 13% 25% --------+----------------+-------- SCC 1558 | 277 1835SCC 1558 | 277 1835 <200 64% | 11% 75%<200 64% | 11% 75% ==================================================== 1838 592 24301838 592 2430 76% 24% 100%76% 24% 100%

Reduced Incidence of NI Rate after Reduced Incidence of NI Rate after 30 days on OmniGen30 days on OmniGen PSCC | PSCCPSCC | PSCC

<200 | >=200<200 | >=200

SCC 212 | 291 503SCC 212 | 291 503 >=200 8% | 11% 19%>=200 8% | 11% 19% --------+----------------+-------- SCC 1702 | 335 2037SCC 1702 | 335 2037 <200 67% | 13% 80%<200 67% | 13% 80% ==================================================== 1914 626 25401914 626 2540 75% 24% 100%75% 24% 100%

Monitoring Fresh Cow Mastitis: Monitoring Fresh Cow Mastitis: Low SCC HerdLow SCC Herd

Cows 5 to 30 days in MilkCows 5 to 30 days in Milk LACT | LACTLACT | LACT

<2 | >=2 All<2 | >=2 All SCC 15 | 27 42SCC 15 | 27 42 >=200 6% | 10% 16%>=200 6% | 10% 16% --------+----------------+-------- SCC 82 | 145 227 SCC 82 | 145 227 <200 30% | 54% 84%<200 30% | 54% 84% ==================================================== 97 172 26997 172 269 36% 64% 100%36% 64% 100%

196,000 SCC196,000 SCC

Monitoring Fresh Cow Mastitis: Monitoring Fresh Cow Mastitis: High SCC HerdHigh SCC Herd

Cows 5 to 30 days in MilkCows 5 to 30 days in Milk LACT | LACTLACT | LACT

<2 | >=2 All<2 | >=2 All SCC 3 | 11 14SCC 3 | 11 14 >=200 5% | 20% 25%>=200 5% | 20% 25% --------+----------------+-------- SCC 9 | 32 41 SCC 9 | 32 41 <200 16% | 58% 74%<200 16% | 58% 74% ==================================================== 12 43 5512 43 55 21% 78% 100%21% 78% 100%

286,000 SCC286,000 SCC

Doing it Right!!!Doing it Right!!!

YIELD & DOLLAR LOSSES

Table 3. Use of DHIA somatic cell count (SCC) score to estimate production loss due to mastitis.

Difference in milk yield* (lb/305 days)

SCC SCS count (1000’s) Lactation 1 > Lactation 2

0 12.5 —— ——1 25 —— ——2 50 —— ——3 100 200 4004 200 400 8005 400 600 12006 800 800 16007 1600 1000 2000

* Comparisons are with lactation yields at SCC scores of 2.Source: Current Concepts of Bovine Mastitis. NMC, 1987.

Lost Production Revenue due to Lost Production Revenue due to MastitisMastitis

Economic loss in milk production due to mastitis compared to a herd with a SCC score of 3

Annualized Loss SCS cents/hd. 500 cow herd 1000 cow herd 3 ----- — — 4 16¢ $24,000 $ 48,000 5 32¢ $48,000 $ 96,000 6 48¢ $64,000 $128,000 7 64¢ $96,000 $192,000

Based on a milk price of $12.00/cwt.

Source: Current Concepts of Bovine Mastitis. NMC, 1987.

Economics of Reduced MastitisEconomics of Reduced Mastitis

100,000 drop in SCC = @15 per cwt. Bonus100,000 drop in SCC = @15 per cwt. Bonus75 lbs. of milk x $0.15 = $11.2 cents75 lbs. of milk x $0.15 = $11.2 cents

Clinical cases of mastitis cost $300 per cowClinical cases of mastitis cost $300 per cow (Guard, et al. 1996)(Guard, et al. 1996)

10% reduction in clinical cases = $10.0 cents per 10% reduction in clinical cases = $10.0 cents per head per dayhead per day

.5 SCS reduction = $8.0 cents in production revenue/day.5 SCS reduction = $8.0 cents in production revenue/day

AbortionsAbortions

Approximately 15% of abortions are mycoticApproximately 15% of abortions are mycotic

The value of a pregnancy is $278.00 (US)The value of a pregnancy is $278.00 (US)

The cost of a pregnancy loss is $555 (US)The cost of a pregnancy loss is $555 (US)

(DeVries 2006; JDS 89:3876 – 3885)(DeVries 2006; JDS 89:3876 – 3885)

Result in Extended LactationsResult in Extended Lactations

Mycotic abortionsMycotic placentitis observed in 131/1107 placentas obtained

Aspergillus hyphae isolated fromplacental tissue (78% incidence)

Phycomycete hyphae isolated fromplacental tissue (11% incidence)

67% of fetal stomachs contained Aspergillus hyphae

McCausland et al., 1997

OR/WA Abortion StudyOR/WA Abortion StudyAbortions

1

10

100

1000

10000

100000

1000000

Cow #20

Cow #63

7

Cow 617

Cow #23

49

Fetus

(617

)

Fetus

(234

9)

Canola

A. fumigatus(spores/ml)

Mycotic Abortions: Field StudiesMycotic Abortions: Field Studies

0

5

10

15

20Ja

n

Fe

b

Ma

r

Ap

r

Ma

y

Jun

Jul

Au

g

Se

p

Oct

No

v

De

c

New crop haylageNew crop haylage

AF addedAF added

Case Study SummaryCase Study Summary Incidence of HBS and Abortion Incidence of HBS and Abortion

----------------Number of Cases----------Number of Cases----------

No OmniGenNo OmniGen11 OmniGenOmniGen

Case No.Case No. Herd SizeHerd Size HBSHBS AbortionsAbortions HBS HBS AbortionsAbortions

112,3 2,3 1200 18 -- 0 -- 1200 18 -- 0 --

2 300 4 -- 0 --2 300 4 -- 0 --

4 350 -- 31 -- 154 350 -- 31 -- 15

5 4500 14 >100 0 05 4500 14 >100 0 0

----------------------------------------------------------------------------------------------------------------------------------------------------------------------------11number of cases reported in the previous 6 month periodnumber of cases reported in the previous 6 month period22binder fed during pre-OmniGen periodbinder fed during pre-OmniGen period33history of vaccination program for clostridium Type Ahistory of vaccination program for clostridium Type A

case locations: 1 & 5; Washington, 2 & 4; Oregoncase locations: 1 & 5; Washington, 2 & 4; Oregon

OmniGen AF - Wisconsin Case StudiesOmniGen AF - Wisconsin Case StudiesMilk Yield & DMIMilk Yield & DMI

No OmniGenNo OmniGen OmniGenOmniGen

NovNov DecDec JanJan FebFeb MarMar AprApr MayMay

Case No. 9*Case No. 9*

Milk (lb) 78 79 84 85 90 90 91Milk (lb) 78 79 84 85 90 90 91

DMI (lb) 48 49 51 51 52 52 53DMI (lb) 48 49 51 51 52 52 53

Case No. 10**Case No. 10**

Milk (lb) 58 60 62 62 62 62 63Milk (lb) 58 60 62 62 62 62 63

DMI (lb) 44 45 50 50 50 50 50DMI (lb) 44 45 50 50 50 50 50

--------------------------------------------------------------------------------------------------------------------------------------------------------------------------

**OmniGen fed at 2oz./h/d beginning after December weigh day, reduced OmniGen fed at 2oz./h/d beginning after December weigh day, reduced to 1oz./h/d mid- Januaryto 1oz./h/d mid- January

**OmniGen fed at 2oz./h/d beginning after December weigh day, **OmniGen fed at 2oz./h/d beginning after December weigh day, reduced to 1oz./h/d at end of Januaryreduced to 1oz./h/d at end of January

In Vitro Mycotoxin Binding Studies with various In Vitro Mycotoxin Binding Studies with various commercial flow agentscommercial flow agents

University of Missouri, ColumbiaUniversity of Missouri, Columbia

% Binding by Other Commercial Products% Binding by Other Commercial Products

Mycotoxin Mycotoxin T-BindT-Bind MycotexMycotex MTB100MTB100 UltraSorbUltraSorb AB20AB20

Aflatoxin 100 99Aflatoxin 100 99 45 10045 100 100100

Fumonisin 100 NT NT NT 100Fumonisin 100 NT NT NT 100Ochratoxin 90 83 81 80 87Ochratoxin 90 83 81 80 87Vomitoxin 30 0 0 13 40Vomitoxin 30 0 0 13 40Zearalenone 61 70 76 100 60Zearalenone 61 70 76 100 60--------------------------------------------------------------------------------------------------------------------------------------------------------------------

Duplicate samples tested with 2 ppm aflatoxin, 2 ppmDuplicate samples tested with 2 ppm aflatoxin, 2 ppm fumonisin, 2 ppm fumonisin, 2 ppm ochratoxin, 4ppm vomitoxin and 2 ppm zearalenoneochratoxin, 4ppm vomitoxin and 2 ppm zearalenone

NT= interference, no results NT= interference, no results

In Vitro Mycotoxin Binding Studies with OmniGen AFIn Vitro Mycotoxin Binding Studies with OmniGen AFUniversity of Missouri, ColumbiaUniversity of Missouri, Columbia

% Binding __________% Binding __________

MycotoxinMycotoxin Sample 1Sample 1 Sample 2Sample 2

Aflatoxin 100 100Aflatoxin 100 100

Fumonisin 83 ntFumonisin 83 nt

Ochratoxin 95 88Ochratoxin 95 88

Vomitoxin 47 55Vomitoxin 47 55

Zearalenone 86 91Zearalenone 86 91

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Duplicate samples tested with 2 ppm aflatoxin, 2 ppm Duplicate samples tested with 2 ppm aflatoxin, 2 ppm fumonisin, 2 ppm ochratoxin, 4ppm vomitoxin and 2 ppm fumonisin, 2 ppm ochratoxin, 4ppm vomitoxin and 2 ppm zearalenonezearalenone

NT= interference, no resultsNT= interference, no results

OmniGen-AFOmniGen-AF

Product category - Nutritional SupplementProduct category - Nutritional Supplement Feeding Rate – 55 grams per headFeeding Rate – 55 grams per head Not sold as a mycotoxin binder …..Not sold as a mycotoxin binder …..

but recent Columbia University but recent Columbia University studies have shown it to be highly studies have shown it to be highly

efficaciousefficacious

ReviewReview All mastitis organisms enter through the streak All mastitis organisms enter through the streak

canal – milking technique, equipment maintenance canal – milking technique, equipment maintenance and hygiene are paramount to low sccand hygiene are paramount to low scc

NEFA & BHB inhibit neutrophil killing abilityNEFA & BHB inhibit neutrophil killing ability OmniGen experimentally increased phagcytosis OmniGen experimentally increased phagcytosis

against against e. colie. coli, increased ROS activity, & , increased ROS activity, & increased nutrophil net formation in vitroincreased nutrophil net formation in vitro

OmniGen-AF reduced SCC in OmniGen-AF reduced SCC in Strep uberisStrep uberis herds herds New infection rate is the driver of SCCNew infection rate is the driver of SCC There may be a good return on investment for There may be a good return on investment for

OAF in some mastitis applicationsOAF in some mastitis applications OmniGen may not be effective against bacteria OmniGen may not be effective against bacteria

forming capsulated cell walls (Klebsiella and forming capsulated cell walls (Klebsiella and Staph)Staph)

Questions?Questions?