Phenotyping and selecting for genetic resistance to gastro-intestinal parasites in sheep: the case of the Manech French dairy sheep breed

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Phenotyping and selecting for genetic resistance to gastro-intestinal parasites in sheep: the case of the Manech French dairy sheep breed JM. Astruc *, F. Fidelle, C. Grisez, F. Prévot, S. Aguerre, C. Moréno, P. Jacquiet * * French Livestock Institute - Toulouse, France Session TS7 Improving Production in Small Ruminants Puerto Varas, Chile, October, 28 th, 2016

The Blond-Faced Manech dairy sheep breed Basque country (south-west of France) : rainy and mountainous area, favorable to gastrointestinal nematodes 280,000 Blond-Faced Manech. Efficient breeding program conducted by CDEO -28% ewes in selection program -150 AI progeny-tested rams / year

< The gastrointestinal nematodes (NGI) parasites in sheep Life cycle Environment (pastures) < Larvae (L3) Eggs > Worms Host (sheep) Abomasum Small intestine Haemonchus contortus Teladorsagia circumcincta Trichostrongylus colubriformis

Why selecting for resistance to NGI parasites? Economic concern Economic losses due to decrease of production and culling Cost of the anthelminthic treatment Increasing resistance to anthelmintic molecules No more effects of molecules in numerous flocks (especially in the Blond-Faced Manech flocks) Environment concern Ecotoxicity => pollution of soil, entomofauna sensitive to chemical residues Genetic selection = sustainable and efficient alternative to treatments?

How to measure resistance to NGI parasites? Host resistance => decrease establishment, development, fecundity and fitness of the worms FEC Faecal Egg Count (eggs per gram) from coproscopy = reference method Also : resilience => maintain performance while subjected to parasite challenge. Measure of packed cell volume (PCV)

How to measure resistance to NGI parasites? Natural infestation on pasture In many countries (Oceania, UK) But : depends on meteorological conditions ; no control of species and larvae ingested Experimental infestation Original design set up in France (Jacquiet at al, 2015) Applied to young rams gathered in breeding centers Future sires Enabled by collective breeding programs Rams must be naïve regarding gastrointestinal parasites

Protocol of experimental infections (Haemonchus contortus) Two periods of infection (duration = 1 month ) Source Jacquiet FEC in eggs / gram PCV in % PCV1i FEC1 (1st infection) PCV1f PCV2i FEC2 (2 nd infection) PCV2f Infection 1 (30 days) 15 days Infection 2 (30 days) D0 D30 D45 D75 Anthelmintic 3500 L3 5000 L3 Anthelmintic

What is the relevance of an experimental infection? High correlation between resistance in experimental infestation vs natural infestation (~ 0.8 to 0.9) High correlation between resistance to Haemonchus contortus vs other species of nematodes (~ 1 Gruner et al. 2004) Correlation between resistance of young rams in breeding center and offspring on pastures is being assessed (on-going on-farm experiments using divergent lines of rams) Resilience (packed cell volume) => allows to check that rams have no pathologic effects

Experimental infections in Manech Blond-Faced 5 experimental infestations design carried out from 2008 to 2015 451 rams, mainly aged 2 or 3 FEC1 FEC2 PCV1f-PCV1i PCV2f-PCV2i Mean 2141 1641 3.4 1.0 Std 2491 1787 3.75 3.15

1 5 9 13 17 21 25 29 33 37 41 45 49 53 57 61 65 69 73 77 81 85 89 93 97 101 105 109 113 117 121 125 129 FEC (number of eggs per gram) Phenotypic variability of rams Infection 2014 : 132 rams in breeding center of Manech blond face (CDEO Ordiarp) 16000 Range of FEC (infestation 2) 14000 12000 10000 8000 6000 4000 2000 0 rams => Important variability between rams

Genetic parameters : traits and model PCV1i Infestation 1 (30d) diffpcv1 = PCV1i PCV1f FEC1 PCV1f 15d PCV2i Infestation 2 (30d) FEC2 PCV2f diffpcv2 = PCV2i PCV2f Model : FEC1 FEC2 diffpcv1 diffpcv2 = + YEAR + AGE + RAM + e REML VCE software All rams from known sire Average : 4.5 rams per sire

Genetic parameters resistance / resilience to nematodes at both infestations h 2 resistance h 2 resilience 0.4 0.4 0.3 0.3 0.2 0.2 0.1 0.1 0 0 FEC1 FEC2 diffpcv1 diffpcv2 ρ g = 0.92 Heritabilities moderate h 2 FEC2> h 2 FEC1. ρ g = -0.49 Surprisingly negative correlation

Genetic correlations between resistance and resilience traits at both infestations ρ(fec1,diffpcv1) ρ(fec2,diffpcv2) -1-0.5 0 0.5 1 1 st infection : resistance and resilience highly correlated 2 nd infection : correlation between resistance and resilience near to zero

Genetic correlation between resistance to nematodes and milk yield (MY) Method of estimation : Genetic evaluation performed on resistance to NGI parasites traits => EBV FEC & REL FEC ρ g = corr(ebv FEC2,EBV MY ) / REL FEC2 xrel MY ρ g FEC2,MY = 0, 184 (unfavorable) Slight unfavorable correlation between resistance at 2 nd infestation and milk yield

Conclusion, perspectives o o o Genetic variability of resistance to nematodes exhibited in an experimental and controlled challenge => selection possible Unfavorable correlation between resistance & milk yield => to be considered in the selection objective Phenotyping resistance to nematodes laborious and expensive. 2 ways to reduce costs work -FEC measure : quantitative real-time PCR from worm DNA currently investigated -Decrease number of individual FEC : measure of FEC2 only (moderate h2, high correlation with FEC1)

Conclusion, perspectives o 2 strategies of selection : Short-term : resistant rams (AI) in flocks with resistance to anthelmintic Long-term : classical selection with selection pressure on rams in breeding center o Genetic evaluation have been performed for 2 years. EBVs provided to Blond-Faced Manech breed society

ICAR issues o Resistance to nematodes in sheep = novel trait o Guidelines could be proposed for recording resistance to nematodes -under natural conditions -under experimental conditions

THANK YOU FOR YOUR ATTENTION Acknowledgements Financial support provided by Aquitaine region (PSDR-INGEDICO program) and FGE (FENOPAR program) Thanks to CDEO (breeding organization of Manech Blond-Faced) and ENVT (Veterinary School of Toulouse) which performed the phenotyping A special thank to Philippe Jacquiet and his team from ENVT