The impact of immune modulation on reproductive performance

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OutlineThe impact of immune modulation on reproductive performance

The connection between immunity and reproduction (inflammation)

Health disorders and reproduction efficiency association (indirect)

Immune cells and reproduction (direct)

OmniGen - relevant research

- Oregon superovulation Snider et. al., 2019

- U of F Casarotto et. al., 2019

- Heat Stress - Florida Fabris et. al., 2017

- Heat Stress - Arizona Hall et. al., 2018

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What's the Immune System all about?Adaptad (Abuelo et al., 2019)

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Rotavirus

BVD virus

Coccidia

Aspergillus

Candida

Tapeworm

Dysfunctional inflammation

Oxidation stress

Altered Nutrient

Metabolism

Metabolic Stress Triad

Inflammation and reproduction

- Energy changes (inflammation), related to Energy

• increase difficulty for cattle to resume estrus

• increase anestrus period

(Crowe, 2008)

Some is normal, prolonged is NOT

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Le Blanc, 2014

“The high fertility cycle”

[Middleton et. al., 2019]

Uterine and other disorders impact repro successRibeiro et al., 2016

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n=5085

RP and Metritis (UTD)

Mastitis, lameness, digestive, respiratory (NUTD)

Inflammatory disease before breeding reduced…

- fertilization of oocytes

- development to morula

- Impaired early conceptus elongation

- Impaired secretion of INF-τ

“Diseases caused inflammation-like changes in the

transcriptome of conceptus cells

- increased risk of pregnancy loss

- reduced pregnancy or calving per breeding”

Health disorders in post-calving dairy cattle…Ingvartsen et. al., 2003

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NAHMS, 2014

40% metabolic

60% infectious

Estrus cycling cows reduced by

- Calving problems 1.9 Xs

- Metritis 2.7 Xs

- Fever 1.8 Xs

Health Problems in the First 60 DIM and Resumption of Estrous Cyclicity in Dairy Cows

Santos et al., 2010

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Category Cyclic, % Adjusted OR (95% CI) P

Healthy 84.1 1.00 ---

1 case of disease 80.0 0.97 (0.72 – 1.30) 0.83

> 1 case of disease 70.7 0.60 (0.44 – 0.82) 0.001

Type of health problem

Calving problem 70.5 0.52 (0.40 – 0.68) < 0.001

Metritis 63.8 0.37 (0.28 – 0.50) < 0.001

Clinical endometritis 68.9 0.51 (0.37 – 0.71) < 0.001

Fever postpartum 80.0 0.55 (0.40 – 0.74) < 0.001

Mastitis 81.5 0.87 (0.55 – 1.36) 0.53

Clinical ketosis 77.7 0.71 (0.47 – 1.07 0.10

Lameness 85.0 0.82 (0.52 – 1.30) 0.40

Pneumonia 88.9 1.78 (0.22 – 14.34) 0.59

Digestive problem 60.7 0.54 (0.25 – 1.17) 0.12

n=5719 postpartum cows, N=7 U.S. dairies

Health Problems in the First 60 DIM and Pregnancy in Dairy Cows Santos et al., 2010

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Category Pregnant, % Adjusted OR (95% CI) P

Healthy 51.4 1.00

1 case of disease 43.3 0.79 (0.69 – 0.91) 0.001

> 1 case of disease 34.7 0.57 (0.48 – 0.69) < 0.001

Type of health problem

Calving problem 40.3 0.75 (0.63 – 0.88) < 0.001

Metritis 37.8 0.66 (0.56 – 0.78) < 0.001

Clinical endometritis 38.7 0.62 (0.52 – 0.74) < 0.001

Fever postpartum 39.8 0.60 (0.48 – 0.65) < 0.001

Mastitis 39.4 0.84 (0.64 – 1.10) 0.20

Clinical ketosis 28.8 0.50 (0.36 – 0.68) < 0.001

Lameness 33.3 0.57 (0.41 – 0.78) < 0.001

Pneumonia 32.4 0.63 (0.32 – 1.27) 0.20

Digestive problem 36.7 0.78 (0.46 – 1.34) 0.38

n=5719 postpartum cows, N=7 U.S. dairies

Pregnancy reduced by

- Calving problems 1.33 Xs

- Metritis 1.5 Xs

- Fever 1.6 Xs

- Ketosis 2.0 Xs

- Lameness 1.75 Xs

Direct

- Neutrophils, macrophages, and T-lymphocytes,

are required by the ovary for normal ovulation

and corpus luteum function

(Walusimbi and Pate, 2013)

Immune System Effects Reproduction

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Indirect

- Impaired neutrophil function causes RPs

(Kimura et al., 2002)

- Impaired neutrophil movement ,

↓phagocytosis - leads to uterine infections

(Lewis, 1997)

Immune Cells recruited to ovulatory follicles

- facilitate ovulation and populate CLs

Active recruitment of immune cells requires

- interaction of immune cells with endothelial cells

- mediated by ligands such as L-selectin, P-selectin,

E-selectin, etc…

- chemotactic proteins (chemokines)

Immune system, essential for optimal reproduction function Walsuimbi and Pate, 2013

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What is IL-8, IL-8R, and L-Selectin?Burton and Erskine, 2003

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Normal immune function Immunosuppression & disease

Cortisol

Immune cells recruited to ovaries during days 1-4 of estrus cycle Jiemtaweeboon et al., 2011

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What is OmniGen-AF?

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A unique, patented nutritional specialty product from

Phibro Animal Health that is recommended to be fed

to dry, pre-fresh and lactating cows every day to help

support healthy immune function in the

face of expected and unexpected stress events

Ingredients

Silicon dioxide, calcium aluminosilicate, sodium

aluminosilicate, brewers dehydrated yeast, mineral oil,

calcium carbonate, rice hulls, niacin supplement, biotin,

d-calcium pantothenate, vitamin B-12 supplement,

choline chloride, thiamine mononitrate, pyridoxine

hydrochloride, riboflavin-5-phosphate and folic acid.

Cortisol

- Adrenal hormone

• Released in response to stressor

• Released by calf, initiates parturition

“Interferes with L-selectin expression

causes neutrophils to lose their

ability to marginate”

OmniGen-AF – restores this function

How does OmniGen-AF work?Burton and Erskine, 2003

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L-selectin

47 2+ Lactation Holstein cows

TX Panhandle

Blood sampled: -60, -30, 0, 14, 28

“OmniGen-AF recovered L-selectin normally suppressed during calving”

How does OmniGen-AF work?Texas Tech (Nightingale et al., 2014)

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Dry Off

-60 DaysCalving DIM: 28

Control n=24

OmniGen-AF 56g/hd n=23

*P < 0.001

*P < 0.002

4 treatments, N=6 Angus Cross cows per treatment – 2 replicates

49 days total feeding OmniGen (0 or 56g/hd/day)

Four day FSH regiment (200mg or 400mg)

Estrous synchronization on Day 28 (Single PGF2α)

Superovulation on Day 38 (after pre-feeding period)

Heat detection 24 h after last PGF

Bred at 0, 12, and 24 hrs after Heat, if no heat by 36 hr post bred 12, 24 hrs

OmniGen-AF® on superovulation response and embryo quality in donor beef cows. Snider et. al., 2019

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Serum collected for [Cortisol] & [Progesterone]

Embryos flushed 7 d after breeding – Counted & Graded 4 rank scheme (Lindner and Wright, 1983)

Statistical analyses were performed using NCSS statistical software, AVNOA 2x2x2 Factorial design.

Significance was declared P < 0.05

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Snider et. al., 2019

[Cortisol]

No difference (P > 0.10) by FSH dose or OmniGen

[Cortisol] less (P <0.05) in Replicate 1 compared to Replicate 2.

Replicate 2, 400mg FSH had the greatest [Cortisol]

0

50

100

150

200

250

300

350

400

450

Day0

Day10

Day14

Day21

Day28

Day38

Day40

Day42

Day43

Day49

Co

rtis

ol (p

g/m

L)

adapted from Snider et al., 2019

Progesterone at embryo collectionSnider et. al., 2019

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Serum progesterone concentration was

significantly higher (P=0.05) in OG fed cows

200mg dose as compared to other treatments.

Elevated P4 likely due to larger/function CL

Number of CL not different

No difference in …

A. recovered OVA between OG and FSH

• Dosage – 1st replicate more OVA

B. number or percentage of embryos recovered

Results recovered ovaSnider et. al., 2019

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Transferrable embryos recoveredSnider et. al., 2019

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Number of transferrable embryos not different

for FSH dose, OmniGen, or replicate

OmniGen fed, 400mg FSH more number (not percent)

transferrable embryos

Number degenerate embryos reduced with OmniGen (P=0.09)

Percentage degenerate embryos reduce with OmniGen (P=0.07)

Degenerate embryos recoveredSnider et. al., 2019

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When using 400 mg FSH regiment OmniGen

- increased transferrable embryo yield

- reduced degenerate embryo recovery

Feed OmniGen, greater progesterone at embryo collection (200 mg FSH)

- may indicate improved luteal function during early embryonic development

Improved transferrable embryo production during superovulation with

OmniGen use

Conclusion

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Effect of feeding an immunomodulator during the dry period and early lactation on health, milk and

reproductive performance in dairy cows

L. T. Casarotto1, J. Laporta1, K. Ferreira1, B. D. Davidson1, K. Moy1, A. K. Almeida1, J.D.

Chapman2, D. J. Mclean2, N. I. Barbu2 V. Ouellet1, and G. E. Dahl1.

1Department of Animal Sciences, University of Florida, Gainesville.

2Phibro Animal Health Corporation, Teaneck, NJ.

1465 multiparous Holstein- cows enrolled July 2018

- Calving occurred from August 2018 through April 2019

- Large commercial herd located near Gainesville, Florida

Cows selected and started diet treatments at dry-off

- N=706 (OmniGen-AF 56g/day) – fed in 0.5 lb allotment

- N=686 (placebo, AB20 control, 56g/day) – fed in 0.5 lb allotment

- Dry-off to 150 DIM or when determined pregnant

Far-off & close-up: Group fed with supplements mixed in TMR

Fresh & Sick: co-mingled with supplements fed as top-dress

Lactating: Group fed with supplements mixed in TMR

Study DesignCasarotto et al., 2019

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Study DesignCasarotto et al., 2019

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Reproduction program

- 77 day voluntary waiting period (VWP)

- Double OvSynch for 1st and 2nd services

- Pregnancy diagnosis using ultrasound @ 30 and 60 days after breeding

- Final preg status at 230 DIM (~80 days AFTER supplementation ended)

Daily Production data obtained by DeLaval parlor milk meters

-Once weekly from 2 through 20 weeks in lactation (U of F)

-Milk composition (Fat, Protein, Lactose, and SCC) was measured every 14 days from weeks 2 -20

Health Disorders

- Detected and recorded (farm staff)

- Dairy Comp 305 records collected weekly (U of F)

OG supplemented cows became pregnant sooner.

OG reduced days open by 10

OG fed cows had reduced average days to

pregnancy (139 d vs. 149 d, P<0.05)

Reproductive performanceCasarotto et al., 2019

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% Pregnant

after

OmniGen-fed Control-fed

1st Service 32.6% 28.1%

2nd Service 52.3% 47.7%

Overall ≤ 230

DIM

76.1% 73.3%

0

10

20

30

40

50

60

70

80

90

100

0 50 100 150 200 250 300P

rop

ort

ion

no

t p

reg

na

nt,

%Days Postpartum

DAYS OPENOG = 139 ± 2.44 d vs. CON = 149 ± 2.69;*P < 0.05

Casarotto et al., 2019

Milk yield and milk compositionCasarotto et al., 2019

* P<0.01OG

DHIA Laboratories

Casarotto et al., 2019

OG tended to reduce ...

- Mastitis (36.8% vs 41.7%., P=0.06)

- Retained placenta (5.44% vs. 7.64%, P=0.09)

Reduced the frequency …

- Metritis, Ketosis, Displaced abomasum,

Digestive disorder, Pneumonia, Lameness

OG reduced average days in hospital

- (6.01 vs 6.56, P<0.01)

Health DisordersCasarotto et al., 2019

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OG was ½ as

likely to have DA

compared to

CON

Impact of Heat Stress

http://www.partners-in-reproduction.com/reproduction-cattle/heat-stress-impact.asp

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Reduced Placental, Fetal and Mammary Development

Heat Stress University of Florida (Fabris et al. 2017)

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Mammary : On day 3, 7, 14, and 27 after dry off HT n=6, CL n=biopsies7, HTOG n=6, CLOG n=5.

-105 Days Dry off -45 Days

Control

OmniGen-AF

“CLOG” n=14

“CL” n=16

“HTOG” n=19

“HT” n=17

60 DIM

45.2 43.6

77.2

69.7

0

20

40

60

80

100

Bre

ath

s p

er

Min

ute

CL

CLOG

HT

HTOG

a a

c b

a,b,c P < 0.05

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Heat StressUniversity of Florida (Fabris et al. 2017)

• Apoptotic rate highest in Cooled OmniGen-AFfed group (p=0.08)

• OmniGen-AF increased alveolar number (p=0.02)

• OmniGen-AF enhances mammary gland restructuring during the dry period associated with improved milk yield?

Heat Stress University of Florida (Fabris et al. 2017)

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0

1

2

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Ap

op

toti

c R

ate,

%

CL CLOG HT HTOG

Apoptosis:

Higher with CLOG

treatment

0

50

100

150

200

Alv

eoli

Nu

mb

er

Control OmniGen-AF

Alveoli:

More with OmniGen-AF

Less with high THI

Heat Stress University of Florida (Fabris et al. 2017)

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40.7 41.1

35.9

40.5

0.0

10.0

20.0

30.0

40.0

50.0

Daily

milk

, kg CL

CLOG

HT

HTOG

a,bP < 0.05

a a a

b

Phase 1: Commercial dairy in Arizona

30 cows (90 DIM)

45 days

Phase 2: Environmental Chambers

12 cows - University of Arizona

- 7 day acclimation (thermoneutral) “TN”

- 10 days heat stress (THI >68) “HS”

- 4 days recovery (thermoneutral) “REC”

[21 days total]

University of Arizona – Heat Stress Hall et al. (2018) JDS: Dr. Bob Collier

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Feed Intake DM, kg. (ARC) Hall et al. (2018) JDS

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Respiration Rate (ARC)Hall et al. (2018) JDS:

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“Thus the negative consequences of postpartum disorders on reproduction

might last up to 10 months after diagnosis and treatment of the problem.

Control of inflammation postpartum using drugs or nutraceuticals seems to

alleviate the negative effects of postpartum disorders on reproduction but

further research is needed to establish firm recommendations. It is

increasingly evident that management for optimal fertility of lactating cows

starts long before breeding, and requires special attention to management of

transition cows.”

Long-term consequences of inflammatory disorders post-partum on fertility #400 ADSA 2019 - Riberio et. al., 2019

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