Effects of feeding intensity and Ca:P ratio on foot welfare in blue foxes (Vulpes lagopus)

Open Jou nal o Animal Sciences, 2014, 4, 153-164 Published Online July 2014 in SciRes. h p://www.sci p.o g/jou nal/ojas h p://dx.doi.o g/10.4236/ojas.2014.44020 How o ci e his pape : Ko honen, H.T., Eskeli, P., Lappi, T., Huuki, H. and Sepponen, J. (2014) E ec s o Feeding In ensi y and Ca:P Ra io on Foo Wel a e in Blue Foxes (Vulpes lagopus). Open Jou nal o Animal Sciences, 4, 153-164. h p://dx.doi.o g/10.4236/ojas.2014.44020 E ec s o Feeding In ensi y and Ca:P Ra io on Foo Wel a e in Blue Foxes (Vulpes lagopus) Hannu T. Ko honen, Pekka Eskeli, Teijo Lappi, Hanna Huuki, Juhani Sepponen MTT Ag i ood Resea ch Finland, Animal P oduc ion Resea ch, Kannus, Finland Email: hannu. .ko honen@m . i Recei ed 25 Ap il 2014; e ised 9 June 2014; accep ed 25 June 2014 Copy igh © 2014 by au ho s and Scien i ic Resea ch Publishing Inc. This wo k is licensed unde he C ea i e Commons A ibu ion In e na ional License (CC BY). h p://c ea i ecommons.o g/licenses/by/4.0/ Abs ac The aim was o cla i y e ec s o ad libi um and es ic ed eeding wi h Ca:P a ios on oo wel a e in blue oxes. Expe imen was s a ed a weaning on Augus 6 h and inished a pel ing on No em- be 26 h, 2013. T ea men s we e: 1) es ic ed eeding, Ca:P a io 1.5:1; 2) es ic ed eeding, Ca:P a io 2.9:1; 3) es ic ed eeding Ca:P a io con ol le el; 4) ad libi um eeding, Ca:P a io 1.5:1; 5) ad libi um eeding, Ca:P- a io 2.9:1; 6) ad libi um eeding Ca:P a io con ol le el. Body g ow h a- iables, eed in ake and wel a e a iables we e measu ed. The beha iou was ideo eco ded. Fo- eleg ca pal join angle as an indica o o leg weakness and he ox’s abili y o mo e we e e a- lua ed. Du ing weeks 32 - 35 and 37 - 47 animals om es ic ed g oups a e 60% - 65% and 67% - 68% o he gi en eed, espec i ely. Body weigh gain was as e in oxes ed ad libi um han wi h a es ic ed die (P < 0.001). A pel ing, wais line and neck ci cum e ence we e signi ican ly (P < 0.001) la ge in ad libi um g oups compa ed o es ic ed ones. The body condi ion sco e was sig- ni ican ly (P < 0.001) la ge o ad libi um animals. Body leng h was g ea e o ad libi um han o es ic ed g oups (P < 0.05). Signi ican di e ences we e no ound in body size a iables be ween Ca:P a ios. Du ing he cou se o expe imen , bending o ca pal join s inc eased in each g oup (P < 0.001). Mos bended join s we e ound a inal sco ing. Signi ican di e ences we e no ed in he angle o join be ween ad libi um and es ic ed g oups (P < 0.05). P onounced di icul ies in lo- como o ac i i y we e no ound. Di e ences be ween he g oups we e sligh in s ick, loo ball and cap u e es s. No pain eac ion was e iden du ing sensi i i y es . Foo wel a e p oblems we e mul i- ac o ial p oblems. Keywo ds Fa med Fox, Animal Wel a e, Fee Condi ion, Feeding Le el, Animal P oduc ion H. T. Ko honen e al. 154 1. In oduc ion Body size o a med blue oxes (Vulpes lagopus) has d ama ically inc eased du ing he las decades. A pel ing, oxes nowadays ypically weigh 15 - 20 kg compa ed o he ime 20 - 30 yea s ago when oxes pel ing weigh was clea ly less han 10 kg [1] [2]. Ad libi um eeding and he high amoun a in he die can pa ly explain he ma ked inc ease o pel ing weigh . Also e ec i e gene ic selec ion has inc eased he body size o animals [3]-[5]. Simul aneously, leg weakness and bending o ca pal join ha e become mo e common [5]. Ob iously, excessi e body mass o size combined wi h such p e-symp oms o os eochond osis may comp omise animal wel a e [1] [6] [7]. The e is a lack o p ope esea ch cla i ying easons causing oo p oblems [8]. Howe e , i can be assumed o be a sum o a ious pa allel ac o s. In addi ion o excessi e body mass and size, o he in luen ial ac o s may be he amoun o calsium (Ca) and phospho us (P) and hei mu ual ela ionships in he die [9] [10]. Resea ch on mine al equi emen o oxes is un o una ely sca ce and was mainly published qui e a long ime ago. S udy by Ha is e al. [11] ecommends ha Ca:P a io should be 1:1.7. A die a y calcium con en below he ecommend- ed le el was ound o cause lameness and bending o leg bones in sil e oxes. Acco ding o he ecommenda- ions based on NRC [12], g owing blue oxes minimum equi emen s o Ca and P in he d y ma e die s a e 0.6% o each. Valaja e al. [13] epo ed ha ash con en o eed 50 g/kg d y ma e and P con en 6 g/kg KA is su icien o sa is y mine al equi emen o g owing oxes. A high amoun o calcium in he die may be also p oblema ic. In dogs, o e eeding caused a highe incidence o ossi ica ion p oblems han es ic ed eeding, and sugges ed ha o e supplemen a ion wi h calcium was a majo con ibu ing ac o [14]. The uppe limi o p op- e calcium supply in oxes is unknown. The aim o he p esen s udy was o cla i y e ec s o wo eeding le els ( es ic ed s. ad libi um) and he h ee Ca:P a ios on oo wel a e in g owing ju enile blue oxes. 2. Ma e ial and Me hods This s udy was ca ied ou a MTT Ag i ood Resea ch Finland, Kannus, in wes e n Finland. The use o expe i- men al animals was e alua ed and app o ed by he Animal Ca e Commi ee o E elä-Suomen aluehallin o i as o (ESAVI/PH 1231 A/2013). The gene al heal h o animals was isually checked daily. 2.1. Expe imen al Se -up and Animals Expe imen al animals we e ju enile male blue oxes bo n in May 2013. Expe imen s a ed a weaning on Au- gus 6 h and inished a pel ing on No embe 26 h. The oxes we e di ided in o six expe imen al g oups: (1) e- s ic ed eeding, Ca:P a io 1.5:1 (R 1); (2) es ic ed eeding, Ca:P- a io 2.9:1 (R 2); (3) es ic ed eeding Ca:P- a io 2.0:1 con ol le el (R 3); (4) ad libi um eeding, Ca:P a io 1.5:1 (AL 1); (5) ad libi um eeding, Ca:P- a io 2.9:1 (AL 2); (6) ad libi um eeding Ca:P- a io 2.0:1 con ol le el (AL 3). The amoun o es ic ed eeding was se o be 60% - 70% o ad libi um le el. Expe imen al eed was manu ac u ed by Te jä F ys eed ki chen. Consump ion o whole expe imen al sea- son was made a once, b ough o Resea ch S a ion, and ozen. The amoun o eed needed o daily consump- ion was smel e e y day. Composi ion o die s is gi en in Table 1 and Table 2. Animals we e ed by hand. Daily eed po ions and le o e s we e weighed wi h a Me le SM 15 balance, accu acy ±1 g. Wa e ing was au- oma ic ad libi um. Animals we e pai -housed in s anda d wi e-mesh cages measu ing 105 cm long × 120 cm wide × 70 cm high. Each cage was u nished wi h a wi e-mesh pla o m (105 cm long × 25 cm wide) loca ed a 23 cm om he ceiling. Fu he mo e, a bone (30 cm long × 7 cm diame e ) as chewing and ac i i y objec was placed in o cage loo . This u nishmen is obliga o y o a med oxes [15]. Animals om same gene ic backg ound (same mo he ) we e e enly dis ibu ed o each s udy g oup. Thus, li e ma es we e placed equally o g oups o homogenize he gene ic a iabili y. 2.2. Body Measu es Body weigh s we e eco ded i e imes du ing he s udy pe iod wi h a Me le SM 15 balance, accu acy ±10 g. A pel ing, body leng h, wais line and neck ci cum e ence we e measu ed by using ape measu e (accu acy ±1 cm). H. T. Ko honen e al. 155 Table 1. Composi ion o expe imen al die s (%). Ca:P 1.5:1 2.9:1 Con ol 2.0:1 Bal ic he ing 25.4 28.1 31.2 Fish o al 5.0 5.0 6.8 Mea and bone meal 1.0 2.1 1.3 Slaugh e house o al; poul y 5.0 17.8 2.7 Slaugh e house o al; bo ine, pig 40.0 23.3 23.6 P o ein admix u e, HP 330 - - 0.02 Fea he meal 0.26 0.24 2.0 Ba ley 3.5 3.5 3.5 Whea 10.0 10.0 10.0 Fox a 3.8 3.6 5.4 Chalk 0.3 - 2.0 DL-me hionine - 0.04 0.06 Phospho us acid (75%) 0.2 - - Wa e 5.4 6.0 11.0 Table 2. Chemical composi ions, and calcula ed con en s o me abolizable ene gy (ME) in he die s. DM = d y ma e . Ca:P 1.5:1 2.9:1 Con ol 2.0:1 DM, % 34.5 36.9 33.1 In DM (%), Ash 8.8 13.4 8.1 C ude p o ein 34.5 29.4 33.0 C ude a 28.2 30.9 27.8 C ude ca bohyd . 31.6 26.3 31.1 ME (MJ/kg DM) P o ein 18.2 18.4 18.0 Fa 27.4 25.0 28.3 Ca bohyd a e 17.4 14.8 17.0 The hickness o subcu aneous a was assessed by a subjec i e body condi ion sco e (BCS) me hod, which gi es an es ima e o he deg ee o a ness independen o he ox’s body size [4]. BCS was e alua ed o each animal on a scale o 1 - 5, whe e 1 = e y hin and 5 = ex emely a . 2.3. Beha iou al Tes s Cap u e es was made be o e weigh ing. Foxes we e caugh wi h neck ongs. Cap u e eac ion was classi ied as con iden o ea ul [16]. The eac ions o he oxes o humans we e e alua ed by pe o ming a walking es [17] [18]. The es was conduc ed ou imes du ing he s udy, i.e. on Aug 9, Oc 1, Oc 29 and No 15. In he es , esea che walked slowly pas he cages and w o e down he beha iou al eac ions. Beha iou s eco ded we e: si ing, s anding, mo ing, lying, on pla o m and s e eo ypy. H. T. Ko honen e al. 156 Explo a i e beha iou was e alua ed wi h a ball es in which a loo ball was placed in he cage. Con ac o ball was eco ded. I he e was no con ac wi hin 10 seconds, he es eac ion was classi ied nega i e. Du ing he s ick es , he esea che was s anding in on o he cage and inse ed a wooden s ick (150 cm long, 5 mm ∅) h ough he wi e-ne 25 cm deep in o he cage. An immedia e eac ion o he s ick s imulus was cha ac e ized as ollows: (1) escape ( ea ul); he animal mo ed away om s imulus; (2) explo a ion (cu ious); he animal app oached he s ick and sni ed and/o bi a he s ick; (3) agg essi eness (agg essi e); he animal made as and in ense a ack a he s ick and main ained he bi e. Sensi i i y o ouch was e alua ed ou imes du ing he cou se o s udy; On Sep 9, Oc 3, Oc 29 and No 26. Sensi i i y es was pe o med by ouching a oo pad sole o ox wi h a ballpoin pen. Foxes’ eac ion o ouch o an objec was immedia ely eco ded. 1 = no eac ion, 2 = mild wi hd awal, sligh pain, 3 = s ong wi hd awal, ma ked pain. 2.4. Beha iou om Video Reco dings The beha iou was moni o ed du ing he expe imen . The ideo eco ding se up comp ised o MIRASYS e- co de (V5016H1), moni o (TFTR 17) and 15 came as (WZI8 PV312-0). The equency o beha iou was ana- lysed om ideo apes using ins an aneous sampling a 1 min in e als o one hou (10:00 - 11:00 am) pe e- co ding day. Reco ding days we e Aug 13, Sep 4, Sep 23, Oc 1, Oc 8, Oc 15, Oc 22, Oc 29, No 5, No 12 and No 19, espec i ely. 2.5. Condi ion o Fee Fo eleg ca pal join angle as an indica o o leg weakness and he ox’s abili y o mo e abou in he cage we e e alua ed bo h li e and om eco dings. E alua ion o ca pal ension was based on subjec i e e alua ion o e- legs [5] by one e alua o on a scale o 1 - 5. In he wo s case (sco e 5 = e y poo ), he ca pal join was bended o a 90o angle compa ed o no mal, only sligh angled ca pal join (Sco e 1 = excellen ). I one o he ca pal join s was mo e ben han he o he , he sco e was based on he wo se leg. Abili y o mo e was e alua ed as ol- lowing [19]. I he e we e no di icul ies o mo e, i was sco ed o be 0. The animal was ac i ely mo ing abou in he cage and was able o jump easily on o he pla o m. Mild di icul ies o mo e we e sco ed as 1. The ox some imes mo ed abou in he cage and was somehow capable o jumping on he pla o m. Ma ked di icul ies o mo e we e sco ed o be 2. The ox emained si ing o lying s ill in he cage, e en when shooed o . 2.6. E alua ion o Fu Quali y Animals we e pel ed (No 26) acco ding o no mal a ming p ac ice. Skin g ading was pe o med by Tu kis ila Luo a Oy. Fu cha ac e is ics e alua ed we e u mass, co e o hai and quali y. The scale anged om 1 (poo es ) o 10 (bes ). Fu de ec s we e also e alua ed. Skins we e weighed wi h a Me le SM 15 balance, accu- acy ±10 g. Skin leng h was measu ed by using a ape measu e, accu acy ±1 cm. 2.7. S a is ical Analyses The s a is ical analyses we e ca ied ou using SAS/STAT so wa e, Ve sion 9.2 o he SAS Sys em o Win- dows. Copy igh © 2009, SAS Ins i u e Inc. SAS and all o he SAS Ins i u e Inc. p oduc o se ice names a e egis e ed adema ks o adema ks o SAS Ins i u e Inc [20], Ca y, NC, USA. 2.7.1. Video Reco dings The o iginal da a consis ed o ele en eco dings (one hou pe day, 10:00 - 11:00 am). Fo he s a is ical analys- es he da a was pooled in o h ee pe iods (pe iod 1: Aug 13, Sep 4, Sep 23, pe iod 2: Oc 1, Oc 8, Oc 15, Oc 22 and pe iod 3: Oc 29, No 5, No 12 and No 19) due o he high a ia ion be ween eco dings wi hin indi- idual animals, especially du ing he las h ee o ou eco dings (pe iod 3). Repea ed measu emen s o ideo eco dings we e analysed using he ollowing linea mixed model: ( ) ( ) ( ) ( ) ijk i j k ijk ij ik jk ijk µ α β γ αβ αγ βγ αβγ ε = Υ ++++ + + + + whe e µ was he gene al mean, αi was he ixed e ec o eeding in ensi y (R o AL), βj was he ixed e ec o H. T. Ko honen e al. 157 Ca:P a io (1.5:1, 2.9:1, con ol le el), γk was he ixed e ec o eco ding pe iod (1, 2, 3), and (αβ)ij, (αγ)ik, (βγ)jk and (αβγ)ijk we e he in e ac ions be ween ixed e ec s. εijk was he esidual e o [21]. Based on he Akaike in o ma ion c i e ion (AIC), he co a iance s uc u e o he epea ed measu emen s was modelled as compound symme y (CS), au o- eg essi e o de 1. (AR1) o uns uc u ed (UN). 2.7.2. Body Size Va iables and Fu P ope ies Measu emen s o body size a iables and u p ope ies we e analysed using he ollowing linea mixed model: ( ) ij i j ij ij µ α β αβ ε Υ= + + + + whe e µ was he gene al mean, αi was he ixed e ec o eeding in ensi y (R o AL), βj was he ixed e ec o Ca:P a io (1.5:1, 2.9:1, con ol le el) and (αβ)ij, was he in e ac ion be ween ixed e ec s. εij was he esidual e o . Since he animals we e pai -housed mean alues o measu ed pa ame e s o animals wi hin each cage we e used in all analyses in all he models. No mali y o esiduals was checked o each analysis using sca e plo s o esiduals and i ed alues. 2.7.3. Ca ego ical Va iables All he ca ego ical a iables we e analysed using he Fishe ’s exac es . 3. Resul s 3.1. Feed Consump ion Feed consump ion in ad libi um g oups (AL 1-3) was signi ican ly highe compa ed o es ic ed ones (R 1-3) (P < 0.001; Figu e 1). A ea ly phase o he expe imen , i.e. on weeks 32 - 35 (Aug 5-Sep 1), animals om e- s ic ed g oups a e 60% - 65% o he amoun gi en o ad libi um animals. Du ing weeks 36-47 (Sep 2-No 24) eed consump ion o es ic ed g oups a e aged 67% - 68% o ad libi um le el. Animals om R 2 and AL 2 g oups had ini ially sligh ly poo e appe i e compa ed o o he g oups. Fu he mo e, AL 2 animals ended o ha e poo e eed consump ion also on la e pa o he s udy, i.e. on weeks 44 - 45 (Oc 28-No 10), and week 47 (No 18 - 24). 3.2. Body Size Ini ial body weigh s o all expe imen al g oups we e simila (Table 3). Howe e , al eady on Sep 9 ad libi um g oups (AL 1-3) e y signi ican ly di e ed om es ic ed eeding g oups (R 1-3) (P < 0.001). This di e ence be ween he g oups emained un il pel ing (No 26). Figu e 1. Feed in ake o expe imen al g oups (g/animal daily) du ing s udy weeks. Week 32: Aug 5 - 11, Week 47: No 18 - 24. H. T. Ko honen e al. 158 Table 3. Body size a iables in exp. g oups. S anda d de ia ions (SD) a e gi en in pa en hesis. P1 = signi icance be ween es ic ed and ad libi um g oups; P2 = signi icance be ween Ca:P le els; P3 = Signi icance be ween hei in e ac ions. *P < 0.05; **P < 0.01; ***P < 0.001. ns = no signi ican di e ence. R1: es ic ed eeding, Ca:P a io 1.5:1; R2: es ic ed eeding, Ca:P- a io 2.9:1; R 3: es ic ed eeding Ca:P- a io 2.0:1 con ol le el; AL1: ad libi- um eeding, Ca:P a io 1.5:1; AL2: ad libi um eeding, Ca:P- a io 2.9:1; AL3: ad libi um eeding Ca:P- a io 2.0:1 con ol le el. Va iable R 1 R 2 R 3 AL 1 AL 2 AL 3 P1 P2 P3 Body weigh s, g Aug 6 5105 5214 5193 5106 5184 5233 ns ns ns (325) (229) (352) (462) (488) (824) Sep 9 7556 7738 7314 9224 8644 8742 *** ns ns (541) (694) (388) (712) (967) (346) Oc 3 9512 9822 9151 12,338 11,890 11,638 *** ns ns (995) (1044) (893) (1142) (1369) (1199) Oc 29 11,230 11,822 10,822 15,556 15,118 14,456 *** ns ns (1391) (1247) (1449) (1430) (1885) (1700) No 26 13,374 14,079 12,775 18,922 17,834 17,316 *** ns ns (1701) (1247) (1449) (1597) (2398) (2213) Body leng h, cm No 26 71.7 73.0 73.0 74.8 73.0 77.3 * ns <0.1 (2.1) (1.7) (2.1) (2.1) (2.8) (2.8) Wais line, cm No 26 65.9 66.4 64.1 80.0 78.3 77.3 *** ns ns (6.1) (4.0) (5.0) (5.1) (6.3) (7.9) Neck ci cum e ence, cm No 26 35.4 36.2 35.5 41.3 41.4 41.0 *** ns ns (2.9) (2.7) (4.0) (2.7) (5.4) (4.4) Body condi ion sco e No 26 3.2 3.3 2.6 4.9 4.8 4.6 *** * ns (0.8) (0.7) (0.5) (0.3) (0.4) (0.7) Wais line and neck ci cum e ence we e signi ican ly la ge in ad libi um g oups compa ed o es ic ed ones (Table 3). Also body condi ion sco e (BSC) was signi ican ly la ge o ad libi um animals. Signi ican di e - ences we e no ound in body leng h a pel ing. Howe e , he e was a clea endency (P < 0.08) ha he oxes in ad libi um g oup we e longe han he ones in es ic ed die g oups. 3.3. Bending o Fee The sco es o he angle o join s in each g oup a e p esen ed in Table 4. In he beginning o expe imen he condi ion o animals’ join s was ound o be ei he good (sco e 2) o excellen (sco e 1). Only ew animals we e classi ied o sco e 3 (su icien ). Ini ially, he e we e no signi ican di e ences in le el o ben join s be ween s udy g oups. Du ing he cou se o expe imen bending o join s inc eased in each g oup. This inc ease be ween ini ial and inal sco ing was signi ican (P < 0.001). H. T. Ko honen e al. 159 Table 4. Bending o ee exp essed as pe cen age o animals ha ing sco e om 1 o 4. Da e Sco e R 1 R 2 R 3 AL 1 AL 2 AL 3 Aug 6 1 0 10 0 0 0 10 2 80 90 80 90 100 80 3 20 0 20 10 0 10 4 0 0 0 0 0 0 Sep 6 1 0 10 0 0 0 10 2 70 70 80 70 80 70 3 30 20 20 30 20 20 4 0 0 0 0 0 0 Oc 10 1 0 0 0 0 0 0 2 40 60 60 30 50 33.3 3 60 40 40 70 50 66.7 4 0 0 0 0 0 0 Oc 29 1 0 0 0 0 0 0 2 20 10 20 0 11.1 33.3 3 20 90 30 40 33.3 33.3 4 60 0 50 60 55.6 33.3 No 18 1 0 0 0 0 0 0 2 20 10 20 0 0 11.1 3 20 80 10 20 44.4 55.6 4 60 10 70 80 55.6 33.3 Mos bended join s we e ound du ing he las sco ing on No 18. Only signi ican di e ences (P = 0.008) he e we e ound be ween R 1, R 2 and R 3 g oups. Pe cen age o animals ha ing sco e 4 (poo ) was 60, 10 and 70%, espec i ely. Fu he mo e, when No 18 da a om ad libi um and es ic ed eeding g oups was pooled oge he , i was ound ha Ca:P le el almos signi ican ly (P = 0.057) explained amoun bending o ee . Du ing No 18, he pe cen age o animals ha ing sco e 4 (= poo ) in g oups wi h Ca:P a ios o 1.5:1, 2.9:1 and con ol we e 70%, 31.6% and 52.6%, espec i ely. Signi ican di e ences we e ound in join bending be ween ad libi um and es ic ed g oups. Nei he signi i- can di e ences be ween ad libi um g oups, no di e ences when No 18 da a om Ca:P le els we e pooled o- ge he was ound. 3.4. Abili y o Mo e Essen ial di icul ies o mo e we e no ound. Thus, animals ha ing a sco e 2 o 3 o he le el o mo ing was no ound un il No embe . Howe e , sligh di icul ies o mo e we e mo e common in ad libi um ed animals compa ed o ones in es ic ed eeding g oup. Pe cen age o animals ha sco ed 1 in g oups R 1, R 2, R 3, AL 1, AL 2 and AL 3 we e 10, 10, 0, 90, 56 44, espec i ely (P < 0.001). Signi ican di e ences we e no ound be- ween AL g oups in abili y o mo e (P = 0.121). 3.5. Beha iou al Reac ions Only ew di e ences we e ound in eac ions o beha iou al es be ween he s udy g oups (Table 5). No di e- ences a all we e ound in s ick and loo ball es . In he cap u e es , howe e , signi ican di e ence was ob- H. T. Ko honen e al. 160 Table 5. Numbe o con iden oxes a cap u e and s ick es s, and explo a i e animals a loo ball es . P = signi- icance be ween all g oups. *P < 0.05; **P < 0.01. ns = no signi ican di e ence. Va iable/Da e R 1 R 2 R 3 AL 1 AL 2 AL 3 P Cap u e es , con iden Aug 6 9 7 10 7 8 9 ns Sep 9 5 4 5 8 10 5 * Oc 3 8 8 5 9 10 8 ns Oc 29 9 3 4 4 6 8 * No 26 6 8 7 8 8 6 ns S ick es , con iden Sep 6 1 0 1 0 1 2 ns Sep 30 0 1 1 0 2 2 ns No 14 0 0 0 0 0 0 ns Floo ball es , explo a i e Sep 6 3 1 4 3 2 2 ns Sep 30 2 1 1 3 4 1 ns No 14 3 1 4 3 2 2 ns se ed in Sep 9 and Oc 29. These di e ences we e no sys ema ic, so, any clea endency o o agains ce ain g oup canno be gi en. No wi hd awal o pain eac ion was ound when es ing sensi i i y o ouch in oxes. All oo pads we e heal- hy in appea ance. Walking es esul s a e shown in Table 6. Only ew di e ences we e obse ed be ween he s udy g oups. Ad libi um ed oxes we e ound o si signi ican ly mo e han es ic edly ed oxes on Oc 29 (P < 0.05). Fu he - mo e, a endency was ound ha si ing was mo e common in ad libi um ed animals on No 15. S e eo ypic be- ha iou was no obse ed du ing any walking es session. 3.6. Video Analyses Da a om ideo eco dings a e p esen ed in Figu e 2. Ac i i y o oxes was highes in Augus and declined o- wa ds pel ing a No embe (P < 0.001). Ac i i y o oxes was same o de o magni ude in all expe imen al g oups. The amoun o si ing was lowes in ea ly Augus and inc eased g adually owa ds pel ing (P < 0.001). Si ing inc eased mo e in he ad libi um ed g oups han in he es ic edly ed g oups (P < 0.05). The amoun o s anding inc eased mo e in he es ic edly ed g oups (P < 0.05). The amoun o lying inc eas- ed in he ad libi um ed g oups while i emained cons an in he es ic ed g oups (P < 0.001). Pla o m use ended o be lowes in ad libi um ed g oups. This endency was mo e clea in la e au umn. 3.7. Fu P ope ies Clea di e ences we e ound in u p ope ies be ween he expe imen al g oups (Table 7). Skin leng h and weigh was g ea e in oxes ed ad libi um compa ed o es ic edly ed ones. Fu he mo e, u mass and u quali y we e be e in ad libi um ed animals. Ca:P le els did no a ec skin weigh , leng h o u mass o ani- mals. The co e o hai and u quali y we e poo es in animals ed 2.9:1 die . 4. Discussion Gene ic backg ound o animals was homogenized. This allowed us o compa e mo e accu a ely e ec s o eed- ing in ensi y and Ca:P die s on oo wel a e. As ou g owing da a showed we success ully p oduced oxes o H. T. Ko honen e al. 161 Figu e 2. Pe cen age o si ing, s anding, mo ing and lying om o al daily ime analysed. No ice he di e - en scaling o y-axis in he g aph illus a ing he si ing beha iou . Video eco ding da es a e gi en in x-axis. Table 6. Beha iou al eac ions du ing walking es . Da a a e gi en as numbe o animals pe o ming he beha- iou . P = signi icance be ween all g oups. *P < 0.05. ns = no signi ican di e ence. Da e Va iable R 1 R 2 R 3 AL 1 AL 2 AL 3 P Aug 9 Si ing 5 6 4 3 4 5 ns S anding 0 0 1 0 0 1 Mo ing 2 2 0 4 0 1 Lying 2 0 1 0 3 0 On pla o m 1 2 4 3 3 2 Oc 1 Si ing 1 4 2 2 5 4 ns S anding 4 0 2 5 2 4 Mo ing 3 2 3 2 1 0 Lying 0 0 0 0 0 0 On pla o m 2 4 3 1 2 1 Oc 29 Si ing 2 3 0 8 5 2 * S anding 2 2 2 1 3 1 Mo ing 1 1 4 0 1 2 Lying 2 3 1 0 0 4 On pla o m 3 1 3 1 0 0 No 15 Si ing 2 4 1 5 5 6 0.06 S anding 3 2 4 5 4 2 Mo ing 1 0 2 0 0 0 Lying 3 1 0 0 0 1 On pla o m 1 3 3 0 0 0