Abstract
Phytoestrogens are plant-produced bioactive secondary metabolites known to play an integral role in plant defense that frequently accumulate in times of stress and/or microbial infection. Phytoestrogens typically belong to two distinct chemical classes; flavonoids (isoflavones) and non-flavonoids (lignans and coumestans). Upon consumption by livestock, high concentrations of phytoestrogens can cause long-term disruption in reproduction due to structural similarities with mammalian estrogens and their tendency to bind estrogen receptors. Wide variation in phytoestrogen concentration has been reported in pasture legumes and corresponding silage or hay. Lucerne is a common perennial pasture legume in temperate climates, but information on phytoestrogen production or accumulation in grazing livestock is currently limited. Therefore, metabolic profiling using UHPLC-MS-QToF was performed to identify and quantitate key phytoestrogens in both fresh and dried lucerne fodder from replicated field or controlled glasshouse environments. Phytoestrogens were also profiled in the blood plasma of Angus cattle grazing field-grown lucerne. Results revealed that phytoestrogens varied quantitatively and qualitatively among selected lucerne cultivars grown under glasshouse conditions. Fresh lucerne samples contained higher concentrations of coumestans and other phytoestrogenic isoflavones than did dried samples for all cultivars profiled, with several exceeding desirable threshold levels for grazing cattle. Coumestans and isoflavones profiled in plasma of Angus heifers grazing lucerne increased significantly over a 21-day sampling period following experimental initiation. Currently, threshold concentrations for phytoestrogens in plasma are unreported. However, total phytoestrogen concentration exceeded 300 mg·kg−1 in fresh and 180 mg·kg−1 in dried samples of selected cultivars, suggesting that certain genotypes may upregulate phytoestrogen production, while others may prove suitable sources of fodder for grazing livestock.
Highlights
Lucerne (Medicago sativa L., known as alfalfa) is a perennial, temperate pasture legume [1] that is widely utilized in mixed farming systems as a preferred source of cattle forage and/or fodder
Molecular features extracted using a non-targeted metabolic profiling approach were subjected to noise removal and normalization followed by targeted identification of key phytoestrogens in lucerne samples
We observed that the concentration of certain flavonoids and other metabolites in lucerne foliar tissues differed between dried samples and fresh samples and was cultivar dependent (Figures 2 and 3)
Summary
Lucerne (Medicago sativa L., known as alfalfa) is a perennial, temperate pasture legume [1] that is widely utilized in mixed farming systems as a preferred source of cattle forage and/or fodder. In comparison to other pasture species, lucerne contains higher concentrations of protein and minerals, while its lower fiber content renders it beneficial for the provision of higher net energy for grazing livestock [2]. Lucerne produces numerous secondary plant metabolites, which may disrupt reproductive processes in livestock, cause bloating and result in reduced palatability. A wide range of the provision of higher net energy for grazing livestock [2]. Metabolites 2021, 11, 55d0uces numerous secondary plant metabolites, which may disrupt reproductive processes in livestock, cause bloating and result in reduced palatability. A wide range of cultivars are available in the Australian market, selected for differing climatic conditions, resistance to pests and/or pathogens [1], seasonality, and suitability for grazing, hay or silage produccutilotinv.aDrseasrpeitaevaaiwlaibdlee isneltehcetiAonusotfralulicaenrnmeagrkeneto,tsyepleecsteredsfuolrtidngiffferroinmglcelgimumateic conditions, breeding andresseilsetcatniocen,tolupceersntseagnradz/eodr lpivaethstoogceknms a[1y],esxehaibsoitnsailgintyi,fiacnadntscuointasbeiqluiteynfcoersgarsa-zing, hay or sociated withsitlhageepprordoduuctcitoinono.f Dbieosapcittievea swecidoendsealreyctmioentaobfolluitceesr,ninecgluendointygppehsorteoscuylttointogxf-rom legume icity and infebrrteileidtyin[g3,a4n].d selection, lucerne grazed livestock may exhibit significant consequences assonsaadmyrsouaennrwmtki-hnseLnmtegleoelsagrwitaloziuismneiamddpnteaeolscaosaadablesyirnsfpunyasosesndntlwttrefuarevtkiocgohnicydeeaLnonnrhgeglnroenaofl-wesievsganwodwatintriizsiuuseattrsmintehimormlrshpludoy(itentlitepoceiahsa.ymbdneesmonenpyo.raz[re,dstnl3eplfytnuav1a,tgersmm4yetaca7noera]nabhrβodctaa.alnivo-owtnroiuaesdinlioncusicrtstittlrdothmtcseeelpierlnuaonsaoseaiaprmnncasdnvtnykoehniezodpenraromwcrytnfnnloleamam)(bteadewnnTa[iytaataa7oltarndsnii]ntrabdsnf.iiodcoddtacosetPrltleeselfeiiepticluevnaphtaslapinsemeaosenneteshsnnvhstaepsystkecdew[pneltec8anritanpodona]geo[n.(.ednny)ga7TwtdseDdis]dprtnd.oininrarsudfsnoPetcooreotaeefyyglldaeistpapeushatunmnterpnoemcoecsentshceeesd[sdsutbys8at[taatspimi]tr5rebintgn.ouugp]inoaugod.ecnc.Ddlis)nlttPinioaututefutprhesntaisorroceetsnrytaaa,tgropatldplhtnoioedcornhgesgtnehceuciryeudsqssmesloctttnuturmae[rrourib5dtuoaluepirusia]ingocsvec.inrltgtentct[oaiPgruiu6tnenttorihar]iperspyeigeanyfiaasthspel[aodltcn6onohortsragd]otheessiyfn,romseottctrouyriqbleotpauisogvrtat[eioro6tnonrix]teygsiiasctea[inin6etrtdoye]ss, (ER) [9,10], smiganmifimcaanltiapnheystotreosgtreongse(ni.ee.x,p1o7sβu-reestirnaldivioels)toacnkdctahnedtreanmdeanticcyalltyo dbiimndineisshtroreg-en receptors productive e(fEfiRci)en[9c,y10[]6,].sPighnyitfioceasntrtopghenytsobeesctraomgeena feoxcpaol saurerea oinf rleivseeasrtochckincathnedlartaem1a9t8i0csa,lly diminish as estrogenicrempertoadbuoclittievsewefefirecifeonucnyd[6to]. hPahveytlooensgt-rsotgaenndsinbgecaadmveersaefiomcaplacatrseaonofborethseaanric-h in the late mal and hum1a9n80rse,parsodeustcrtoivgeensyicstmemetsa[b6o]l.iLteivsewsteorcekf,opuanrtdicutolahralyvcealtotlnega-nstdanshdeienpg, agdravzeirnsge impacts on legume pastbuoreths awniitmh ahliagnhderhpuhmyatonersetprorogdenucctoivnetesnytstheamvse[b6]e.eLnivsehsotwocnk,topaerxtipceurliaernlycecartet-le and sheep, productive dgyrasfzuinngctlieognumraengpiansgtufrreosmwiltohwheigrhceornpcheypttoioenstrroagteesn caonndteinntcrheaavseedbeeemn sbhroywonnicto experience loss [11], to croemprpoldeutectinivfeerdtiylistfyu[n9c]t.ion ranging from lower conception rates and increased embryonic
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