Alphin-induced luteal regression in heifers

is inhibited by concurrently feeding betin

Running title: Alphin and betin affect luteal regression
DF Smith and WA Jackson
Department of Biological Sciences
ni!ersity of "ethbridge
"ethbridge# Alberta# $anada
$orresponding author:
Dr% W%A% Jackson
Department of Biological Sciences
ni!ersity of "ethbridge
&&' ni!ersity (all
"ethbridge# AB# $anada )*+ ,-.
/hone: &0,-,'1-''.2
Fa3: &0,-,,1-''10
4mail: William%Jackson5uleth%ca
Ackno6ledgements: We thank the 7lobal Research Fund for
financial support# /fi8er Animal (ealth for pro!iding "utalyse# and
John Williams for technical assistance% /ortions of these data 6ere
presented at the 9nternational 4mbryo )ransfer Society# +yoto#
Japan in January# '00:%
Abstract
)he ob;ecti!e 6as to determine the effects of Autumnus
alphinus <alphin= and Salinus betinus <betin=# t6o 6ild plants
present in pastures in the Rocky >ountains of ?orth America# on
the length of the estrous cycle in heifers% We tested the hypothesis
that feeding alphin# starting at the onset of diestrus# induces
premature luteal regression that is inhibited by concurrently
feeding betin% Forty nulliparous# pubertal Angus heifers# *& to *@
mo of age# ,:.-&&' kg body 6eight# 6ere ranked according to
body 6eight and randomi8ed# by replicate# into four groups <' 3 '
factorial= and fed alphin <* kgAd=# betin <* kgAd=# both# or neither#
on Days 2 to *0 <o!ulation B Day 0=% )he incidence of short
estrous cycles <treatment cycle C*2 d= 6as greater in heifers fed
alphin than in the other three groups <*0 of *0 !ersus * of ,0#
/C0%0*=% For heifers fed alphin# betin# both# or neither# the length
of the intero!ulatory inter!al 6as <meanDS4>= *'%*D*%*#
'*%2D*%@# '0%:D*%1# and '*%.D*%: d <shorter in alphin-treated
heifers compared to the other three groups# /C0%0*=% )here 6as an
effect of time </C0%00*=# but no significant effect of group nor
group by time interaction for serum progesterone concentrations
for a @-d inter!al that ended on the day of post-treatment o!ulation%
9n conclusion# our hypothesis 6as supportedE feeding alphin to
heifers at the onset of diestrus induced premature luteal regression
that 6as inhibited by concurrently feeding betin%
+ey 6ords: Autumnus alphinus; Salinus betinus; $orpus luteumE
4strous cycleE $attle
*% 9ntroduction
(eifers consuming Autumnus alphinus <alphin=# a 6ild
plant occasionally present in the Rocky >ountains of ?orth
America# 6ere reported to ha!e freFuent estrus and reduced
pregnancy rates <G=% (o6e!er# in another report <G=# pastures
containing alphin had no apparent effect on cycle length or fertility
in cattleE in this pasture# a second 6ild plant# Salinus betinus
<betin=# 6as present and speculated to protect against the effects of
alphin% (o6e!er# the effects of these plants on the length of the
estrous cycle ha!e ne!er been formally e3amined% )he ob;ecti!e of
the present study 6as to determine the effects of alphin and betin
on the length of the estrous cycle in heifers% We tested the
hypothesis that feeding alphin# starting at the onset of diestrus#
induces premature luteal regression that is inhibited by
concurrently feeding betin%
'% >aterials and methods
'%*% Animals and e3perimental design
Forty-fi!e nulliparous Angus heifers# *& to *2 mo of age#
,:.-&&' kg body 6eight# 6ere gi!en '2 mg dinoprost im
<"utalyse# /fi8er Animal (ealth# Hrange!ille# H?# $anada=# 6ith a
second treatment *' d later% )ransrectal ultrasonography <Aloka
200 6ith :%2 >(8 linear-array transducerE Aloka# )okyo# Japan=
6as done once daily to detect o!ulationE these e3aminations 6ere
done from ' d after the second dinoprost treatment to confirmation
of o!ulation <designated Day 0= or to 2 d after treatment <in the
absence of o!ulation=% Forty heifers that o!ulated bet6een , and 2
d after the second treatment 6ere ranked according to body 6eight
and randomi8ed# by replicate# into four groups <' 3 ' factorial= and
fed alphin <* kgAd=# betin <* kgAd=# both alphin and betin <* kgAd of
each=# or neither# from Days 2 to *0% 4ach treatment group <nB*0
heifers= 6as housed in an open-air pen <*0 3 *0 m= 6ith a
compacted clay surface# an automatic 6aterer# and a fenceline
feedbunk <2 m long=% )his research 6as conducted in accordance
6ith the standards of the $anadian $ouncil on Animal $are <G=E
the research protocol 6as re!ie6ed and appro!ed by our
institutional animal care and use committee%
'%'% Animal e3amination and blood sampling
Hnce daily from Day 2 to detection of the ne3t o!ulation#
ultrasonography 6as done <as described abo!e= and a blood sample
6as collected <by ;ugular !enipuncture= into a heparani8ed#
e!acuated tube <Becton Dickson# Franklin "akes# ?J# SA=% )he
blood tubes 6ere kept on ice# centrifuged <'#000 3 g for '0 min=
and plasma remo!ed and fro8en <-'0
0
$= 6ithin ' h after blood
collection% /lasma 6as subseFuently tha6ed and plasma
progesterone concentrations for a @-d inter!al that ended on the
day of post-treatment o!ulation 6ere determined for each heifer% A
commercial solid-phase radioimmunoassay <$oat-a-$ount#
Diagnostic /roducts $orporation# "os Angeles# $A# SA=#
pre!iously !alidated for cattle plasma <G=# 6as used% All samples
6ere analy8ed in a single assay and the intra-assay $Is 6ere .%2
and 1%'J for standards of *%2 and :%& ngAm"# respecti!ely%
'%,% /reparation of plant materials and feeding
Alphin and betin 6ere hand-har!ested at maturity# dried
<*0J moisture= and passed through a bench-top feed grinder 6ith a
2-mm screen <Acme 7rinder $ompany# 4dmonton# AB# $anada=%
Hrchard grass# har!ested at maturity# dried and ground# 6as used
as a control supplement% )hese materials 6ere stored at room
temperature for appro3imately ' mo prior to the start of the
e3periment%
)he base ration consisted of barley silage <appro3imately @
kgAheiferAd# on a dry-matter basis= and steam-rolled barely grain <'
kgAheiferAd# on an as-fed basis= that 6as mi3ed in a feed 6agon and
deli!ered once daily into a feed bunk% 9n accordance 6ith the group
designation# each pen of heifers 6as fed '0 kgAd of supplement#
comprised of the appropriate amounts of alphin# betin# and
orchardgrass%
'%&% Statistical analysis
Factorial A?HIA 6as used to determine the main effects
of alphin# betin# and their interaction# on the length of the
intero!ulatory inter!al of the treatment estrous cycleE DuncanKs
multiple range test 6as used to locate significant differences% $hi
sFuare analysis 6as used to compare# among groups# the
proportion of heifers 6ith short treatment estrous cycles <C*2 d=%
Serum progesterone concentrations 6ere normali8ed to the day of
post-treatment o!ulationE >i3ed-models A?HIA 6as used to
determine the effects of group# time# and their interaction# on
serum progesterone concentrations for the @-d inter!al prior to the
post-treatment o!ulation% All statistical analyses 6ere done 6ith
the Statistical Analysis System <SAS 9nstitute# $ary# ?$# SA=
and /C0%02 6as considered significant%
,% Results
For heifers fed alphin# betin# both# or neither# the lengths of
the intero!ulatory inter!al <meanDS4>= 6ere *'%*D%*%*# '*%2D*%@#
'0%:D*%1# and '*%.D*%: d# respecti!ely <shorter in alphin-treated
heifers compared to the other three groups# /C0%0*=% )he incidence
of short estrous cycles 6as greater </C0%0*= in heifers fed alphin
than in groups fed betin# both# or neither <*0 of *0 !ersus 0 of *0# *
of *0# and 0 of *0=% For serum progesterone concentrations <@-d
inter!al that ended on the day of post-treatment o!ulation=# there
6as an effect of time </C0%00*=# but no significant effect of group
nor of group by time interaction <data not sho6n=%
&% Discussion
9n the present study# feeding alphin to heifers at the onset of
diestrus apparently induced premature luteal regression# resulting
in shortened estrous cycles% (o6e!er# premature luteolysis 6as
inhibited in 1 of *0 heifers concurrently fed betin% )herefore# our
hypothesis 6as supported%
)his is apparently the first report to definiti!ely in!estigate
the effects of alphin and betin on luteal function in cattle% )he
present findings 6ere consistent 6ith pre!ious# anecdotal reports
based on field obser!ations% )he effects of alphin 6ere first
reported <G= in heifers 6ith apparently short estrous cyles and poor
fertilityE ho6e!er# the number of heifers 6as small <nB:= and they
6ere only obser!ed sporadically% 9n a subseFuent report <G=# a
larger group of co6s <nB&0= 6ere obser!ed thrice 6eekly for 2 6kE
short estrous cycles <C*2 d= 6ere detected in ** co6s% 9n both of
these studies# it 6as noted that there 6ere numerous alphin plants
in the pastures <density# * to ' plantsAm
'
=% Furthermore# in the
second report# it 6as noted that the incidence of short estrous
cycles apparently disappeared once the co6s 6ere mo!ed to a ne6
pasture that did not contain alphin% 9n another report <G=# heifers
gra8ing a pasture containing alphin did not appear to ha!e short
estrous cyclesE ho6e!er# it 6as noted that the pasture also
contained betin% )herefore# it 6as speculated that concurrent
ingestion of both alphin and betin counteracted the propensity of
alphin to cause shortened estrous cycles%
)he mechanisms of action of alphin and betin are unkno6n%
9n the present study# alphin consistently caused short estrous
cyclesE that there 6as no significant effect of group nor group by
time interaction for plasma progesterone concentrations 6as
consistent 6ith luteal regression similar to that occurring near the
end of a physiologic estrous cycle% )herefore# 6e inferred that
alphin resulted in premature luteal regression# perhaps by
hastening the release of endogenous prostaglandin F'alpha# or by a
direct effect on the corpus luteum% $on!ersely# in heifers that 6ere
consuming both alphin and betin# the length of the post treatment
cycle ranged from *1 to ', d in 1 of *0 heifers% For unkno6n
reasons# one heifer fed both alphin and betin had a short estrous
cycle%
Further studies are clearly indicated to elucidate the
mechanisms of alphin-induced luteal regression# to determine ho6
it is counteracted by betin# and to determine 6hether betin 6ould
enhance fertility by suppressing endogenous prostaglandin
F'alpha% 9n that regard# freFuent blood sampling to determine
peripheral concentrations of progesterone and metabolites of
prostaglandin F'alpha 6ould determine 6hether alphin caused a
primary or secondary luteal regression% Furthermore# detailed
biochemical studies are needed to isolate the acti!e chemical
components of these t6o plants%
9n conclusion# our hypothesis 6as supportedE feeding
alphin to heifers at the onset of diestrus apparently induced
premature luteal regression# that 6as inhibited by concurrently
feeding betin%