Postnatal Development of the Urine Cortisol/Creatinine Ratio in the Dog

Pos na al De elopmen o he U ine Co isol/C ea inine Ra io in he Dog K. Vos a KoVá1,4, P. Š a ha2, P. Kaláb3, J. blahoVá1, E. ba anyioVá1 1Depa men o Ve e ina y Public heal h and oxicology, 3Depa men o biophysics and biochemis y, Facul y o Ve e ina y hygiene and Ecology, Uni e si y o Ve e ina y and Pha maceu ical sciences, b no, Czech epublic 2Depa men o Compu e G aphics and Geome y, Facul y o Mechanical Enginee ing, b no Uni e si y o echnology, Czech epublic 4) Ve e ina y Clinic E ika, P ague, Czech epublic Recei ed Augus 16, 2007 Accep ed Ap il 14, 2008 Abs ac Vos a ko á K., P. Š a ha, P. Kaláb, J. blaho á, E. ba anyio á: Pos na al De elopmen o he U ine Co isol/C ea inine a io in he Dog. ac a Ve . b no 77, 2008: 509-513. he objec i e o his expe imen was o s udy he age changes o co isol o c ea inine a io in u ine o g owing puppies be ween bi h and he age o 8 weeks. beagle puppies (n = 14) o h ee li e s we e used. he puppies o li e s 1 and 2 we e om a esea ch b eeding acili y and li e 3 om a dog b eede . hey we e nu sed by hei dams and ob ained ex a puppy ood om he 4 h week o age. he puppies we e weighed weekly. Mo ning u ine was sampled a he age o up o 48 h, and 7, 14, 21, 24, 35, and 56 days. U ine co isol was measu ed using he ia me hod, and c ea inine was de e mined using a diagnos ic ki . he u ine co isol o c ea inine a io (UCC ) was calcula ed. he body mass o puppies inc eased mo e han en imes. he UCC o li e s 1 and 2 as agains li e 3 showed no signi ican di e ences and he e o e he esul s we e pooled. Thus no e ec o he ea ing en i onmen upon his indica o was ound. he UCC alues in puppies aged up o 48 h was 44.32 ± 30.47 × 10-6. i s ollowing changes (a dec ease o 30.52 ± 12.98 × 10-6 on day 7 and oscilla ions a ound hese alues un il day 49) as well as an inc ease on day 56 we e no signi ican . Cha ac e is ic ea u es o he UCCR in puppies we e alues su passing hose epo ed o heal hy adul animals, and a g ea in e -indi idual a iabili y, p esumably e lec ing he ma u a ion p ocesses such as apidly inc easing body mass and skele al muscula u e mass, changes in nu i ion, locomo o y ac i i y and ma u a ion o hei exc e o y sys em. The speci ic ea u es o his pe iod should be conside ed when e alua ing he UCC in puppies. Age changes Beagle, g ow h, body mass, u ine, co isol, c ea inine Changes in co isol sec e ion ha e epea edly been measu ed as a esponse o a ious s esso s in animals (e.g. Kojima e al. 1995, boissy and le neind e 1997). since blood collec ion i sel is o en s ess ul o animals, non-in asi e sample collec ion me hods ha e been de eloped and alida ed. hey only minimally dis u b he animals and enable o assess hei co isol concen a ions in sali a and u ine, and ha e also been used in dogs when s udying hei wel a e issues o diagnosing diseases (bee da e al. 1996, 1999ab; bee da e al. 2000, s olp e al. 1983). When u ine is used o analysis, quan i a i e measu emen o i s co isol exc e ion can be de e mined o a 24 h u ine collec ion pe iod. howe e , an al e na i e o 24-h collec ion is he de e mina ion o he u ine co isol o c ea inine a io (UCC ) in a andom sample o u ine since c ea inine exc e ion is expec ed o be ai ly cons an du ing 24-h pe iods in heal hy animals (Finco 1997). hus he a io o co isol and c ea inine in he u ine can be used o accoun o he e ec o u ine concen a ion. a simple s ess, such as a ca ide o isi o a e e ina ian may cause a mild inc ease in his a io ( an Vonde en e al. 1998; uo salo and an 2004). in adul undis u bed (non-s essed) beagle dogs he UCC alues be ween 6.2 ± 1.0 × 10-6 and 17.8 × 10-6 we e ound (e.g. Jones e al. 1990; bee da e al. 1996; s ephen and ledge 2006). al hough a ious aspec s o enal unc ions du ing pos na al de elopmen ACTA VET. BRNO 2008, 77: 509-513; doi:10.2754/a b200877040509 Add ess o co espondence: E. ba anyio á Dep . o Ve e ina y Public heal h and oxicology, FVhE Uni e si y o e e ina y and Pha maceu ical sciences Palackého 1-3, 612 42 b no, Czech epublic Phone: + 420 541 562 791 E-mail: [email p o ec ed] h p://www. u.cz/ac a- e /ac a e .h m o dogs we e s udied in some de ail, such as body compa men s and Pah clea ance in neona al puppies and egula ion o hei enal unc ions (Čapek and Helle 1961; Helle and Čapek 1965), we ound no da a on UUCR in puppies. I has been known ha hei neph ogenesis is incomple e a bi h, and hei glome ula il a ion a e (GFR) inc eases be ween days 2 and 77 se en old (ho s e and Val in 1971, in Finco 1997). lane e al. (2000) pe o med a quan i a i e u inalysis in beagle puppies aged 9 o 27 weeks, and quali a i e u inalysis in di e en b eeds o puppies aged 0-8 weeks was p esen ed by Faulks and lane (2003); howe e , no UCC was included in hese s udies. We he e o e decided o s udy he de elopmen o co isol/c ea inine a io in puppies aged 0 o 8 weeks a e bi h. Ma e ials and Me hods a o al o ou een newbo n beagle puppies o h ee li e s we e included in he s udy. wo li e s (n = 8, 3 males and 5 emales) we e si ed by he same male, bo n and ea ed in a esea ch acili y and kep a oom empe a u e o 26.5 °C and ela i e humidi y o 38%, na u al dayligh . beginning om week 4, he puppies we e o e ed addi ional puppy ood. hey we e dewo med on day 1 (P obican, Medipha m), 26 (D on al Junio baye , Ge many), and 35 (Caniquan el plus, heinz haup Pha ma Gmbh, Ge many). Puppies o li e 3 (n = 6, 2 males and 4 emales) we e bo n and ea ed in he household o a dog b eede , and ea ed in a sepa a ed kennel oom ca 3  3 m, a an ambien empe a u e o ca 25 °C du ing he i s h ee weeks. They we e dewo med a he ages o 2, 4, 6 and 8 weeks (Banmin h pas e, P ize AH, Belgium). All puppies we e weighed a weekly in e als. U ine samples we e collec ed in he mo ning be ween 07.00 and 09.00 h. in he neona al pe iod, u ine was oided a e a gen le massage o he inguinal egion o puppies o e a clean plas ic ay and collec ed using a sy inge. in olde puppies, u ine oided a e hei waking in he mo ning was immedia ely collec ed om he clean conc e e loo using a sy inge. All samples we e hen placed in Eppendo ials and ozen o -20 °C wi hin an hou o collec ion pending analysis. Co isol in u ine was de e mined using a ia ki (immuno ech P ague, a beckman Coul e Company) ca . 1841. analysis was pe o med using a di ec me hod wi hou ex ac ion. C ea inine concen a ion was de e mined using he diagnos ic ki bioVendo lK ea inin. Fo s a isical e alua ion o he da a, s uden ’s - es , pai ed - es and Kolmogo o -smi no es we e used. Resul s and Discussion he mean body mass o puppies inc eased om 303  68 g on he day o bi h o 3508  42 g on day 56, i.e. mo e han en old. his g ow h a e was simila o da a desc ibed o labo a o y beagle dogs by Field and Jackson (2006). We ound no signi ican di e ences in UCCR (pai ed - es ), when puppies o li e s 1 and 2 we e compa ed wi h hose o li e 3, he e o e he da a o all puppies we e pooled. hus no e ec o he ea ing en i onmen upon hei UCC was ound in he ea ly pos na al pe iod. he mean UCC in puppies aged up o 48 hou s was 44.3  30-47 × 10-6. howe e , because he UCC alues ound o he indi idual days did no show a no mal dis ibu ion (as es ed by he Kolmogo o -smi no es ), only a possible de elopmen endency could be desc ibed (see Fig. 1). he e was a g ea indi idual a iabili y in he UCC in he pe iod unde s udy. also plasma c ea inine concen a ion in dogs seems o be subjec o g ea in e -indi idual a iabili y as ound in beagle dogs (Cen e e al. 1985; b aun e al. 2003). Kuhl e al. (2000) ound highe plasma c ea inine in o weile puppies han in beagle puppies. in gene al, he UCC alues in ou puppies su passed subs an ially hose epo ed o adul and heal hy dogs o he same b eed. in adul beagle dogs ha ha e no been exposed o any s ess he UCC o 6.2 ± 1.0 × 10-6 and 11.9 ± 1.2 × 10-6 in dichlo ome hane ex ac ed and nonex ac ed u ine, espec i ely, we e ound (bee da e al. 1996). simila basal UCC , namely, 9.4 × 10-6, was epo ed by Jones e al. (1990), and a median alue o 17.8 × 10-6 by s ephen and ledge (2006) o kennelled dogs. simila median alue, namely 10.48 × 10-6, was ound in en heal hy adul dogs using a ia me hod (Kole ská and s oboda 2000). Median alues in ou puppies oscilla ed be ween 28.29 × 10-6 and 71.40 × 10-6. When e alua ing he p esen esul s, he speci i ies o he de eloping kidney ha e o be 510 aken in o conside a ion. Quali a i e u inalysis was pe o med in young puppies (Faulks and lane 2003), and he au ho s s a ed ha i can be in e p e ed as o adul dogs in puppies olde han ou weeks. Glucose and p o ein concen a ions app oach he alues ound in adul dogs by 3 weeks o age (bo ee e al. 1984, Fe man and allen 1991). howe e , no c ea inine concen a ion was included in hese s udies. Quan i a i e s udies o blood se um c ea inine showed a dec ease in he i s week o li e in Beagle puppies wi h a s able concen a ion up o he end o he second mon h wi h no di e ences be ween males and emales (Kuhl e al. 2000). no sex di e ences in UCC we e ound in ou puppies. in se e al s udies using o he animal species i was shown ha he kidney is physiologically p og ammed o depend on glome ula and ubula ac ions o p os aglandins in he pe ina al and neona al pe iod (heymann 1987). i can be assumed ha he widely sca e ed alues o he UCC in ou s udy we e due o mo phological and unc ional imma u i y o he canine kidney bu may also be connec ed wi h changes in c ea ine and c ea inine syn hesis in he phase o apid body and muscula u e mass inc ease ( he puppies inc eased hei body mass mo e han en old in he pe iod unde s udy). in his con ex i is impo an o men ion he indings o a signi ican ly highe se um c ea inine concen a ion in heal hy g eyhound dogs as compa ed o non-g eyhound dogs (D os e al. 2006). he au ho s asc ibe his di e ence o he la ge muscle mass in g eyhounds. Fu he mo e, locomo o ac i i y du ing he pos na al pe iod a ec s he ac i i y o c ea ine kinase as shown in a s (Demin and and oso a 1994) and i may be assumed ha i makes i sel el also in he syn hesis o c ea ine and c ea inine. no doub , he p o ound changes aking place du ing he i s 8 weeks o pos na al li e o puppies, om o al nu i ional and he mo egula o y dependence upon hei mo he du ing he neona al pe iod, when he puppies ba ely mo e, o ansi ional pe iod (days 14-28) wi h opening o he eyes and ea s, beginning o u ina ion and de eca ion wi hou s imula ion by hei mo he , apidly inc easing locomo o ac i i y (houp 1998, bea e 1999), documen ed e.g. by a mo e han wo old and signi ican (p  0.01) inc ease in ho izon al locomo o ac i i y du ing a 15-min es (Vos a ko á 2008) be ween days 35 and 56 a e bi h, a ec all body sys ems, including he de elopmen o endoc ine egula ions. i can be specula ed ha he wo old ( hough no signi ican ) inc ease in UCCR be ween days 49 and 56 may be connec ed wi h he abo e-men ioned changes. 511 Fig. 1. UCC o puppies in he ea ly pos na al pe iod (maximum alues, qua ile 75%, median = ho izon al ba , = a i hme ic mean, qua ile 25%, minimum alues) + Ma os e al. (1998) obse ed ubula eabso p ion o c ea inine in he newbo n abbi , also in p ema u e and e m in an s (Coul ha d 1985). hey p opose ha his is due o a slow u ina y low along he ubule and in pa icula due o inc eased back-leak o c ea inine along he imma u e ubula s uc u es. i can be said ha ubula eabso p ion o c ea inine is a physiological phenomenon in he imma u e de eloping kidney. Plasma c ea inine doubled in he newbo n abbi , he u ina y exc e ion o c ea inine mus ha e almos doubled as well due o unchanged slow u ina y low (Ma os e al. 1998). In conclusion, speci ic pos na al de elopmen s o hese unc ions we e p esumably in ol ed also in ou expe imen al animals. in his s udy o UCC in puppies aged 1 o 56 days, alues oscilla ing be ween 30.52  3.91  10-6 and 44. 32  8.80  10-6 wi h a non-signi ican inc ease o a double alue o 91.20  17.45  10-6 on day 56 we e ound, along wi h conside able in e -indi idual a iabili y. hese alues we e highe han hose epo ed o adul indi iduals o he same b eed. The speci ic ea u es o his pe iod should be conside ed when e alua ing he UCC in puppies. Pos na ální ý oj pomě u ko isolu a k ea ininu u š ěňa Cílem s udie bylo sledo a ěko é změny pomě u ko isolu a k ea ininu (UCCR) moči os oucích š ěňa od na ození do ěku osmi ýdnů. Použi a byla š ěňa a plemene bígl (n = 14), ze ří hů. V hy 1 a 2 byly odcho á ány e ýzkumném zařízení, h 3 u cho a ele. Všechna š ěňa a byla kojena a od 4. ýdne přik mo ána. Jednou ýdně byla ážena. Ranní moč byla odebí ána e ěku do 48 h, a po é 7., 14., 21., 35. a 56. den po na ození. Koncen- ace ko isolu moči byla měřena pomocí RIA me ody, ke s ano ení k ea ininu byl pou- ži diagnos ický ki . Z ýsledků byl ypočí á án pomě ko isolu a k ea ininu. Hmo nos š ěňa z os la pokusném údobí íce než 10 . V pomě u ko isolu a k ea ininu nebyly mezi hy nalezeny ozdíly, p o o byly šechny hodno y sloučeny. Vli ůzného p os ředí se edy na UCCR nep oje il. UCCR š ěňa do 48 h po na ození byl 44,32  30,47  10-6. Následující změny (pokles na 30,52  12,98  10-6 do 7. dne a podobné hodno y do 49. dne), podobně jako zes up 56 dne ýznamné nebyly. Cha ak e is ickým znakem UCCR š ěňa byly hodno y přesahující nálezy u dospělých psů a elká indi iduální a iabili a, p a děpodobně e lek ující p ocesy ma u ace jako jsou ychlý ůs ži é hmo nos i a kos e - ní s alo iny, změny ýži y, pohybo é ak i i y a dalších unkcí. 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