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ContentsTwenty ejaculates from _ve dairy AI!bulls were used to compare\ in a split!sample experiment\ the fertility ð45 day! non!return!rate "NRR# from more than 03999 AI# and sperm viability post!thaw of semen diluted with an egg yolk! "Tri! ladyl þ # or soybean!based "Biociphos!Plus þ # commercial extender[ The in vitro evaluations were divided in two experi! ments[ Experiment 0 "n 19# included post!thaw evaluations of motility "subjective and computerized#\ membrane integrity "CalceinAM:EthD!0\ SYBR!03:PI\ and osmotic resistance testÔ RT#\ and capacitation status "CTC:EthD!0#[ Experiment 1 "n 09# included evaluations of the capacitation!"CTC:EthD! 0# and acrosome status "FITC!PSA:EthD!0# during incubation with:without a challenge with solubilized zona pellucida pro! teins "SZP#[ No signi_cant di}erence in the fertility "58[0 2 9[7 versus 58[1 2 9[7# results was found between the two extenders[ In experiment 0\ the computerized motility evaluations post! thaw "CASA# showed higher values for Biociphos!Plus þ pro! cessed semen for the velocity patterns and lateral sperm head displacement[ After 5 h at room temperature "19Ð11>C# all the CASA motility patterns were signi_cantly higher for Biociphos! Plus þ [ The proportion of spermatozoa with intact membranes assessed by CalceinAM was signi_cantly higher in Biociphos! Plus þ "p ³ 9[990# compared to Triladyl þ \ but such di}erence was not seen when using SYBR!03 or the ORT!assay[ When using the CTC:EthD!0 assay\ a lower proportion of acrosome reacted "AR# spermatozoa post!thaw "p ³ 9[90# was found in Biociphos!Plus þ processed semen\ as well as a tendency "p ³ 9[96# for a higher number of uncapacitated spermatozoa[ In experiment 1\ the proportion of uncapacitated spermatozoa was signi_cantly higher for Biociphos!Plus þ when semen was incubated "27>C and 4) CO 1 # without SZP at both 9 "p ³ 9[990# and 29 min "p ³ 9[94#[ Concomitantly\ Triladyl þ showed a higher percentage of capacitated spermatozoa at 9 "p ³ 9[90#\ 29 "p ³ 9[94# and 019 min "p ³ 9[94#[ A higher "p ³ 9[94# incidence of AR!spermatozoa was seen in Triladyl þ at the beginning of the incubation with SZP[ No signi_cant di}erence between extenders was detected for the acrosome status by the FITC!PSA!assay[ Incubation with SZP induced acrosome reaction of capacitated spermatozoa in both extenders\ which was detected by CTC and FITC!PSA assays[ In conclusion\ fertility was not a}ected by Biociphos!Plus þ when 04 × 09 5 of spermatozoa per AI dose were inseminated[ The _nding that higher frequencies of spermatozoa seemed more membrane stable post!thaw\ when frozen in Biociphos! Plus þ \ might indicate that this extender better protects the sperm viability compared with Triladyl þ [