Transition cow health and immune function
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1 Transition cow health and immune function Ynte Schukken, Brianna Pomeroy and Anja Sipka Cornell University Wageningen University Utrecht University GD Animal Health
2 Introduction Transition cow health: Transition from lactation to dry Transition form dry to lactation Transition from healthy to diseased Transition from diseased to healthy Resilience Mammary microbiome Immune responsiveness
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5 What is milk microbiome? Microbial populations as found using next generation sequencing or pyro-sequencing. Population of bacterial species identified using 16S sequencing of bacterial DNA isolated from milk Approximately 7000 bacterial 16S sequences corresponding to ~1000 bacterial species per milk sample (1 ml), and likely more in milk from healthy quarters.
6 Number species in milk Mammary microbiome Healthy Quarters T. pyogenes Mastitis quarters Number of sequences in milk Oikonomou et al. 2014
7 T. pyogenes+ samples % of sequences 1 Arcanobacterium Porphyromonas Fusobacterium Oikonomou et al. 2016
8 SCC <20 aerobic culture negative 80 % of identified sequences Oikonomou et al. 2016
9 S. uberis+ samples 1 % of identified sequences Streptococcus
10 Fusobacterium Genera level results: Culture-negative mastitis samples Asticcacaulus % of identified sequences
11 Culture Negative CM samples versus SCC <10K 100% 90% 80% 70% 60% 50% 40% 30% 20% 10% 0% % of identified sequences
12 Self vs non-self Gensollen et al. 2016
13 Farm specific mammary eco system Farm I Farm II Contributing: anaerococcus, streptococcus, propionibacter,..
14 Is there a resilient microbiome? Patho-gen Non-gen Physio-gen Sphingobacterium Streptococcus Paenibacillus Anaerococcus
15 Ecologic resilience Scheffer et al. 1993
16 Challenge infection - E. coli - LV RV Control LA RA Challenge Control Antibiotics Ganda et al. 2017
17 100% 50% 0% 100% 50% 0% 100% 50% 0% 100% 50% 0% Time (hours) Ruminococcaceae Enterobacteriaceae Aerococcaceae Lachnospiraceae Corynebacteriaceae Planococcaceae Bacillaceae Clostridiaceae Bacteroidaceae Staphylococcaceae Micrococcaceae Paraprevotellaceae Pseudomonadaceae Succinivibrionaceae Spirochaetaceae Prevotellaceae Rikenellaceae Xanthomonadaceae Veillonellaceae Streptococcaceae Fusobacteriaceae Peptostreptococcaceae Weeksellaceae S24-7 Comamonadaceae Other
18 Treatments Challenge infection - E. coli Enterobacter (%) E. coli challenge * * * * * * * *
19 Treatments Challenge infection - E. coli Shannon Index of diversity E. coli challenge Challenged, Non-Treated Challenged, Treated * * * *
20 Antibiotics in healthy quarters Shannon Index of Diversity Not challenged, not treated Not challenged, treated Treatments
21 Intramammary resilience Scheffer et al. 2001
22 Microbial eco-systems Take home messages The mammary gland of dairy cows harbors a microbial ecosystem. Potential for a resilient ecosystem Intramammary challenge results in a loss of bacterial diversity. Some cows are more resilient to exotic bacterial growth compared to other more susceptible cows. Immediate appropriate antibacterial treatment results in a reduced loss of diversity.
23 Dry period immunobiology Minor Staph. Strep. E. coli IMI in late gestation No IMI in late gestation Green et al. 2002
24 Challenge in late gestation: E. coli Parturition 100 cfu E. coli Quesnell et al. 2012
25 Pro-inflammatory Cytokine Il-1 : no response Calving Days after challenge 100 cfu E. coli Clinical mastitis challenge strain Quesnell et al. 2012
26 IL-4 (ng/ml) Anti-inflammatoire Cytokine response: IL cfu E. coli hour periods relative to challenge Gurjar et al. 2013
27 Pro-inflammatory response Cytokine: IL-1 Mid-lactation Hours post challenge 100 cfu E. coli Sipka et al. 2013
28 Th1:Th2 ratio Mid-lactation Post-partum Late gestation
29 Improve immune response Innate: Teat-end quality Lactoferrin NEB Immune modulation: Selenium/vitamin E Cytokines Adaptive: Vaccination E. coli & S. aureus & other Imm immunostimulation
30 Incidence Clinical Mastitis Bovine Granulocyte Colony Stimulating Factor (rbg-csf) 40% 35% 30% 25% 20% 15% 10% 5% 0% Saline 5 μg/kg 10 μg/kg 20 μg/kg Dose Peg - BGCSF Treatment N N % mastitis Duration Severity cows mastitis Saline % ,95 5 μg/kg % 5.3 3,2 10 μg/kg % 8.4 3,3 20 μg/kg % Hassfurther et al. 2015
31 E. coli vaccination reduces severity 32 E. coli challenge study 30 Milk Production (Kg) Control Vaccinated ~ 50% reduced production loss Days relative to challenge Wilson et al. 2006
32 N=10 N=5 Intramammary immunization with Days -44, UV IR E. coli -30, -16 Vaccine J5 Bacterin N=10 Day -30 LF LH RF RH IMM sensitization w/ equivalent 100 cfu E. coli C1 Days -44, -30, -16 Day -30 Vaccine J5 Bacterin N=5 LF RF LH RH Day 0 immuniza IMM sensitization w/ tion w/ equivalent 100 cfu E. coli C1 Control N=5 IMM challenge w/ 100 cfu/ml E. coli C1 Control blank IMM sensitization immunization Control PBS IMM challenge Day 0 IMM challenge w/ 100 cfu/ml E. coli C1 Control blank Pomeroy et al. 2016
33 Probability of having IMI Log 10 (CFU+1) Intramammary immunization with UV irradiated E. coli 100 live CFU P -10d Parturition Control Immunized 100 CFU UV IR E. coli Pomeroy et al. 2016
34 Pregnancy and Immune response Immune response in late gestation: anti-inflammatory IL-4 spike No pro-inflammatory response after challenge Immune response changes from late gestation to early lactation: From anti-inflammatory to pro-inflammatory Infection in late gestation but clinical response in early lactation..
35 Ecologic resilience in transitions Scheffer et al. 1993
36 Take home message Mammary microbiome High diversity is important Resilient microbiome is likely Farm specific Treatment affects microbiome Immune responsiveness Late gestation affects immune bias Systemic and local immune response may be improved Resilient system prevents infection and disease
37
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