EAAP 213 Nntes Frnce Horse Commission Session Do stllions recognize the estrous stte by smelling the odor of mres? C Brint (1), A Boukkz (2), Y Gudé (3), I Couty (4), D Guillume (4), JM Yvon (3), Y Murin (5), O Rmpin (5), B Nielsen (5), M Mgistrini (4) (1) Institut Frnçis du Chevl et de l Equittion, Hrs Ntionl de Blois, 4143 Blois Cedex (2) Lbortoire d'hygiène et de snté nimle, Institut Ntionl des Sciences Vétérinires, Université Ibn Khldoun, Tiret, Algérie (3) INRA, Unité Expérimentle de Physiologie Animle de l Orfrsière, 3738 Nouzilly (4) INRA, UMR Physiologie de l Reproduction et des Comportements, 3738 Nouzilly, (5) INRA Unité Neurobiologie de l Olfction et Modélistion en Imgerie, 78352 Jouy en Joss.
In severl mmmlin species odornt signls re given off by estrous femles MICE Urine of proestrous/estrous femles Attrction of the mle (Achirmn nd Archunn 26) RAT Feces of estrous femles Erections in mles (Rmpin et l 26) ELEPHANT Urine of the estrous odor femles of mre in estrus Flehmens = lip curl in mles (Rsmussen et l 1982, 1996, 1997, 21) Introduction AND IN HORSE? Horses could percept hlf mile wy??? COW Cervico-vginl mucus, urine, feces, sliv Attrction of the mle, flehmens nd mount (Klemm et l 1987, Snkr et l 27, Rmesh Kumr et l 2, Snkr et Archunn 28) HOWEVER Sight > olfction > hering (Blschke et l 1984) Bulls do not show preference with estrous/diestrous restrined femles (Wllch nd Price 1988) or when physicl contct is denied (Gery et l 1991)
Question 1 : Is there n estrous odor in equine species? YES! Men number of erections presented by rts when smelling odor of feces from RAT FOX HORSE Negtive controls FAECES 3 species Rt Fox Horse 3 sttes Estrous femle Diestrous femle Mle Estrus Diestrus Mle Brint odeurs sexuelles Brint EAAP 213 Estrus Diestrus Mle Figure I Estrus Diestrus Mle (Rmpin et et l l 26)
Question(s) 2 : Do stllions recognize the estrous stte by smelling the odor of: A.Mres? B. Urine of mres? C. Feces of mres? Brint EAAP 213
A. Do stllions recognize the estrous stte by smelling the odor of mres? 17 m (55 feet) Stllion N=6 Two-choice tests : estrous mre / diestrous mre 5 Estrous mre Trnquillized mres Acepromzine + Romifidine Without Without Without sight sight olfction 5 Diestrous mre Ech test Ech experiment Control test/tretment test Repeted on 2 successive weeks Alternting the sexul sttes of mres 3 experiments (= 3 tretments) 1) Without olfction 2) Without sight 3) Without rectivity of mres = trnquillized mres Auditory stimuli lwys present
A. Control test :full behviourl sequence with the estrous mre Sniffing, licking, nuzzling from the hed to the til Interctions with perinel region Flehmen(s) Attempt of mount Brint EAAP 213
A. Control test Seconds Behvior of the Stllion With the estrous mre With the diestrous mre Totl time spent ner ech mre Totl time of of sniffing interctions licking bitting with perinel region 2 8 15 * 6 * 1 4 5 2 Seconds Nb Behvior of the mres Estrous mre Diestrous mre 12 8 4 Number of cceptnce behviors * Seconds 15 1 5 Totl time of ttempts to mount NS Seconds 12 9 6 3 Totl time of flehmens * Nb 2 15 1 5 Number of refusl behviors * Brint EAAP 213 * = P <.5 (n=6) (Brint et l, 21)
A. Without olfction Stllions still show preference for the estrous mre The behviour of the mres is not modified: Acceptnce for the estrous mre Refusl for the diestrous mre Only the totl time of flehmens is reduced Brint EAAP 213 To detect the estrous mre ofction is not bsolutely necessry Another role? (Brint et l, 21)
A. Without sight Among the 5 stllions 5 tried to find the mre with sight Brint EAAP 213 (Brint et l, 21)
A. Without sight Among the 5 stllions only 1 find the mres with olfction.but did not discriminte the estrous mre! Brint odeurs sexuelles To detect the estrous mre sight is bsolutely necessry nd olfction lone is not sufficient (Brint et l, 21)
Seconds A. Trnquilized mres Behvior of the Stllion With the estrous mre With the diestrous mre Totl time spent ner Totl Totl time time of of sniffing interctions licking ech mre Conclusion bitting with perinel region 1 2 12 8 15 NS 6 NS 8 1 4 4 5 its rectivity 2 is essentil To detect the estrous mre Seconds Nb Behvior of the mres Estrous mre Diestrous mre Number of cceptnce behviors NS Seconds 15 1 5 Totl time of ttempts to mount NS Seconds 12 9 6 3 Totl time of flehmens NS Nb 2 15 1 5 Number of refusl behviors * Brint EAAP 213 * = P <.5 (n=6) (Brint et l, 21)
B. Do stllions recognize the estrous stte by smelling the odor of urine of mres? Sequences sniffing / flehmen Swb (1) Sniffing (2) Flehmen Urine of pro-estrous mre Urine of estrous mre Urine of diestrous mre Urine of stllion 2 negtive controls Beginning of nsl dischrge Disinterest behviours Brint EAAP 213
B. Odor of urine of mres 14 12 1 8 6 4 2 Durée Totl séquences time of sequences reniflement sniffing/flehmen / flehmen b c OEST PROE DIEST MALE VAN PHY 14 12 1 8 6 4 2 Beginning Début écoulement of nsl dischrge nsl b b OEST PROE DIEST MALE VAN PHY Mens +ET 1 Number Nombre of ctions disinterest de desinteret behviours c Stllions n=6 8 6 b 4 2 Brint EAAP 213 OEST PROE DIEST MALE VAN PHY Brint, not published
B. Odor of bovine urine Urine estrous mre 18 12 6 Urine Cow or bull Totl time of sequences Durée séquences reniflement / sniffing/flehmen flehmen estrus EQ estrus BOV Negtive control b b Estrous Estrous Diestrous Bull Control phy mre cow cow diestrus BOV mâle BOV Mens +ET Stllions n=6 25 2 15 1 Début écoulement nsl Beginning of nsl dischrge b Brint odeurs sexuelles 5 Estrous estrus EQEstrous estrus BOV Diestrus diestrus mâle Bull BOVControl phy BOV mre cow cow Brint, not published
Conclusion 2 Sniffing of urine Stllions : - Recognize the odor of urine - Do not seem to recognize the odor of estrus - Discriminte between the odor of mre nd the odor of stllion - Recognize bovine urine = common molecules? Brint EAAP 213
C. Do stllions recognize the estrous stte by smelling the odor of feces of mres? Experiment 1 Behviour of stllions Hormones Experiment 2 Behviour of stllions Semen collection Qulity of semen Brint EAAP 213
C. Odor of feces of mres? Experiment 1 6 Welsh pony stllions 4 feces odors - Estrus - Diestrus - Mle - Negtive control Sniffing of feces for 5 min Brint EAAP 213 Blood smples From 3 min to + 2h fter sniffing of feces Assys Testosterone nd prolctin
Experiment 2 C. Odor of feces of mres 4 Welsh pony stllions 4 feces odors Sniffing of feces for 5 min Semen collection Semen qulity Immedite survivl fter 24 nd 48 h: filtered volume, gel volume, concentrtion, totl sperm count, sperm motility pure nd diluted in INRA 96 Brint EAAP 213
C. Odor of feces of mres Behviour : sniffings Mens +ET Experiment 1 Experiment 2 Temps (Time) sec 2 15 1 5 Estrus P =,1 Diestrus Stllion Control Oestrus DIestrus Mâle Témoin Temps (Time) sec 2 15 1 5 Estrus DiestrusStllion Control b b Oestrus Dioestrus Mâle Témoin Stllions recognize feces Brint EAAP 213 Brint et l, 211
C. Odor of feces of mres Behviour : Flehmens Mens +ET Experiment 1 Experiment 2 Temps (Time) sec 6 5 4 3 2 b 1 b Oestrus DIestrus Mâle Témoin Estrus Diestrus Stllion Control Temps (Time) sec 6 5 4 3 2 1 b b b Oestrus Dioestrus Mâle Témoin Estrus Diestrus Stllion Control Stllions discriminte between stllion nd mre feces Brint EAAP 213 Brint et l, 211
C. Odor of feces of mres Hormones Testosterone Mens concentrtions Before/After sniffing * Prolctin Estrus Diestrus Stllion Control Estrus Diestrus Stllion Control Testosterone = indictes the recognition of the odor of «mle»? Prolctine = indictes the recognition of the odor of «femle»? * : p<.5 Brint EAAP 213 T : p<.1 Brint et l, 211
C. Odor of feces of mres Semen : totl time of semen collection 1 Mens +ET Stllions n=3 Temps (Time) sec 8 6 4 2 Estrus Diestrus Stllion Control Oestrus Dioestrus Mâle Témoin Reduction of the totl time of semen collection with feces of mre? Semen qulity No effect could be seen Brint EAAP 213 Brint et l, 211
Conclusion 3 Sniffing of feces Stllions : - Recognize the odor of feces - Do not sem to recognize the odor of estrus - Discriminte between the odor of stllion nd the odor of mre Testosterone nd prolctin Could indicte this recognition Feces of mres (estrus or diestrus) could llow quicker semen collection Brint EAAP 213
THANKS to The tem of experimentl studs of INRA in Nouzilly Guy Duchmp, Fbrice Reigner, Philippe Brrière, Jcky Mérigrd, Frnçois Stiu, Thierry Blrd L Institut Frnçis du Chevl et de l Equittion For the finncil support