Diuretic Effects of Several Chemical and Herbal Compounds in Adult Laying Hens

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1 Interntionl Journl of Poultry Siene 9 (3): , 2010 ISSN sin Network for Sientifi Informtion, 2010 Diureti Effets of Severl Chemil nd Herl Compounds in dult Lying Hens Esfndiry, H. Rjin, K. ssi, M. nsri Lri nd J. Jlei 1 Shool of Veterinry Mediine, Shirz University, Shirz, Irn 2 Deprtment of Bsi Sienes, Shool of Veterinry Mediine, Shirz University, Shirz, Irn Deprtment of Clinil Sienes, Shool of Veterinry Mediine, Shirz University, Shirz, Irn 4 Deprtment of Food Hygiene, Shool of Veterinry Mediine, Shirz University, Shirz, Irn 3 strt: Diureti gents hve een used in the tretment of severl situtions suh s rdiopulmonry nd urinry diseses in humns nd nimls, ut they hve not een extensively studied in poultry. Some Irnin poultry produers trditionlly use urotropin (hexmine), lhgi (Cmelorum fish) nd mlt everge to get therpeuti effet in roiler or lying hens. Diuretis my e used fter drug therpy in order to get rid of tissue residues nd in some toxiity onditions in irds. The purpose of this study ws to evlute the diureti properties of severl herl nd hemil gents in dult lying hens. dult lying hens were rndomly divided into 9 groups, eh group onsisting of 5 irds. Speifi doses of diuretis were dministrted to nesthetized hens (with xylzine nd ketmine) following 24 h off-feed period. Urine smples were olleted for 2 h fter drug dministrtion. Blood smples were lso tken t 90 min. The onentrtions of hloride, lumin, gluose, retinine, uri id nd ure nitrogen were mesured in serum nd urine smples. Urine volume nd speifi grvity, sodium nd potssium onentrtions were lso determined in urine. Sttistil nlysis of dt showed tht furosemide hd positive signifint effet on urine volume ompred to other gents. Urotropin lso inresed the urine volume ut this effet ws not signifint. With respet to the effets on eletrolytes (sodium, potssium nd hloride), intrmusulr furosemide, orl urotropin nd hydrohlorothizide exerted some hnges, ut no onsistent pttern ws oserved with respet to the other ftors mesured in serum nd/or in urine. Diureti effet ws more pronouned following intrmusulr dministrtion of furosemide. Key words: Diureti effet, herl nd hemil ompounds, lying hens INTRODUCTION Unlike the lssil en-shped kidneys in mmmls, kidneys in irds re longitudinl in shpe nd re omprised of three loes with no distint ortex nd medull (Brun, 1982). The funtioning units of eh kidney re nephrons nd these enle the irds to sve their ody wter y onentrting the urine. The terminl olleting tuules empty into the ureters whih diretly open in the lo, ommon vestiule into whih the digestive nd reprodutive trts end. Therefore, no renl pelvis nd ldder is present in irds (Diker nd Hslm, 1966; Lverty nd Dntzler, 1982). Birds mke urine hving one fifth onentrtion ompred to tht in most mmmls (Brun nd Dntzler, 1972; Emery et l., 1972; Lyton et l., 2000). Uri id is the min nitrogenous wste produt exreted lrgely y tuulr seretion. Uri id in the urine is present in supersturted olloid form suspension nd does not ontriute to the osmoti pressure (MN, 1974; MN nd MN, 1975). Diuretis re sustnes tht use n inrese in exretion of eletrolytes nd wter so tht ody gets rid of ny exess fluid. Their mehnisms of tion vry nd generlly involve speifi enzymes, trnsport proteins, hormone reeptors nd ion hnnels funtioning diretly or indiretly in renl tuulr eletrolyte resorption (Brter, 1998; Pushett, 1981). Osmoti diuretis, however, exert their effets y estlishing n osmoti pressure grdient etween plsm nd tissue wter omprtments (Kng nd Kim, 2005). Diuretis re used in severl rdiovsulr disorders suh s ongestive hert filure, hypertension, hyperklemi, sites, ertin kinds of tissue edem nd high intrrnil pressure (Czj et l., 1983; Dormns et l., 1998; Rodés, 1996; Roudeush et l., 1994; Vestweer et l., 1989; Widemn et l., 1995). These gents my lso e used in ertin types of toxiosis onditions in nimls nd humn (Berg, 1977; Roselli nd Yu, 1970; Selye, 1969; Selye et l., 2006). Diureti ompounds hve not een extensively studied in poultry. Trditionlly, some Irnin poultry produers use urotropin, lhgi nd mlt everge during het stress periods nd fter vintion of irds, or fter n outrek of infetious diseses nd they lim to get positive therpeuti effets minly due to their diureti tion. So, the im of this study ws to exmine the diureti effet of those gents nd severl other hemil nd herl produts in lying hens. Corresponding uthor: H. Rjin, Deprtment of Bsi Sienes, Shool of Veterinry Mediine, Shirz University, Shirz, Irn 247

2 MTERILS ND METHODS nimls: Fourty five 60 week-old Bovns lying hens weighing kg were purhsed from Veterinry Shool Edutionl Frm of Shirz University nd were sujeted to nine groups nd were treted ording to the followings. Hens were kept in 15 ges with similr onditions of environmentl temperture, humidity, lightening nd nutrition. Birds fed d li nd hd free ess to drinking wter. The feed ws deprived to the hens 24 h efore the experiment in order to redue the mount of intestinl ontents, ut they hd ess to wter during this period. Chemil nd herl produts: Drugs were purhsed from lol medil or veterinry drugstores. Herl produts were otined from trditionl produers in Shirz. ll solutions were freshly prepred from solid drug dosge forms. Drug dministrtion to vrious groups: Nine groups of hens were treted s the following: C Control group for orl route (10 ml kg distilled wter dministered diretly into the rop y plsti feeding tue). C Control group for prenterl route (0.25 ml kg distilled wter injeted intrmusulrly into petorl musle). C Mlt everge group (10 ml kg ; given orlly). C lhgi group (10 ml kg ; given orlly). C Orl furosemide group (5 mg kg ; from 0.05% drug solution fter dissolving furosemide tlets in deionized wter). C Urotropin (methenmine; hexmine) group (50 mg kg ; from 0.5% solution of the powder; given orlly). C Hydrohlorothizide group (10 mg kg ; from 0.1% drug solution fter dissolving hydrohlothizide tlets in deionized wter; given orlly). C Spironoltone group (1 mg kg ; from 0.01% drug solution fter dissolving spironoltone tlets in deionized wter; given orlly). C Prenterl furosemide group (2.5 mg kg ; from 1% drug solution; given intrmusulrly). nesthesi: Ketmine hydrohloride (12.5 mg kg ) nd xylzine hydrohloride (5 mg kg ) were injeted intrmusulrly to indue generl nesthesi. Smll dose of ketmine hydrohloride (5 mg kg t most) ws re-dministered to mintin nesthesi when required. Eh nesthetized ird ws pled y its k on surgil ord with the nek nd wings extended. The legs were fixed to the ord with msking tpe. Blood smpling: The site of leeding ws prepred y removing fethers nd disinfeted with 70% ethnol. Blood smples were tken from wing veins t 90 min fter nesthesi. Smples were olleted in tues not ontining ny kind of ntiogulnt gent nd were left t room temperture for 2 h nd ser were then seprted y entrifugtion t 1500g for 20 min. Gluose ws mesured right fter lood entrifugtion. The remining ser were divided into severl tues nd o were kept t -20 C until nlysis for lumin, retinine, uri id, lood ure nitrogen (BUN) nd hloride. Urine olletion: Fethers were removed to expose the lol region nd the re ws disinfeted. smll piee of guze lurited with minerl oil ws inserted in the oprodeum of lo to prevent urine ontmintion with intestinl ontents. Urine smples were intermittently olleted, using plsti syringe, from the fluid ppering in oprodeum. Eh time the olleted urine ws emptied into tue nd ws ultimtely entrifuged t 2500 g for 10 min. fter entrifugtion, the volume of the superntnt ws mesured nd liquots were divided into plsti vils o nd kept t -20 C until nlysis. Gluose ws mesured immeditely fter finl urine olletion. Other urinry iohemil prmeters were mesured t the end of in vivo experiments. nlytil proedures: Gluose ws mesured with enzymti-olorimetri (gluose oxidse) method (Brhm nd Trinder, 1972). Serum nd urine lumin onentrtions were determined using romresol green ssy (Doums et l., 1971; Tietz, 1976). BUN nd UUN were mesured y dietyl monoxime tehnique (Evns, 1968). Uri id ws determined y phosphotungstte method in oth urine nd serum smples (Reee nd Hoie, 1972). Urinry nd serum retinine were estimted y the Jffe retion (Brtels et l., 1972; Finy nd Ertingshusen, 1971). Colorimetri thioynte method ws used to determine hloride onentrtions in oth serum nd urine smples (Tietz, 1976). Flme photometry ws employed to mesure the onentrtions of sodium nd potssium in urine (Cmpell nd Coles, 1986; Dein, 1986). Speifi grvity of urine ws determined y refrtometry (Cmpell nd Coles, 1986; Dein, 1986). Sttistil nlysis: Results re expressed s Men± SEM. unless otherwise stted. NOV test ws used to nlyze the dt. The level of signifine ws onsidered to e p<0.05. RESULTS () Urine physiohemil hrteristis: The dt with respet to the levels of vrious urinry prmeters fter dministrtion of different ompounds in hens re summrized in Tle

3 Tle 1: Levels (Men± SEM; n = 5) of urinry exretion of vrious prmeters during 2 h fter dministrtion of different gents in lying hens DW (PO) DW (IM) F (IM) F (PO) HCT (PO) Ftors (units) (10 ml/kg) (0.25 ml/kg) (2.5 mg/kg) (5 mg/kg) (10 mg/kg) Volume (ml) 4.5± ± ± ± ± ± ± ±6.4 0±0 0±0 Chloride (meq/l) 7.0± ± ± ± ±1.2 lumin (g/dl) 0±0 0±0 0±0 0±0 0.04±0.03 Cretinine (mg/dl) 29.8± ± ± ± ±1.6 Uri id (mg/dl) 23.5± ± ± ± ±2.4 Ure nitrogen (mg/dl) 6.3± ± ± ± ±2.1 Sodium (meq/l) 365±32 447± ± ± ±37 Potssium (meq/l) 386±61 309±40 608±86 657±98 457±51 Speifi grvity (mg/ml) ±0.1 NM** ± ± ± SPL (PO) UT (PO) L (PO) MB (PO) Ftors (units) (1 mg/kg) (50 mg/kg) (10 ml/kg) (10 ml/kg) Volume (ml) 2.3± ± ± ± ±3.8 B 12.5± ±8.6 0±0 Chloride (meq/l) 4.2± ± ± ±2.0 lumin (g/dl) 0±0 0±0 0±0 0±0 Cretinine (mg/dl) 26.3± ± ± ±2.3 Uri id (mg/dl) 18.8± ± ± ±2.2 Ure nitrogen (mg/dl) 17.0± ± ± ±4.3 Sodium (meq/l) 361±61 433± ± ±701 Potssium (meq/l) 406±56 553±86 716± ±393 Speifi grvity (mg/ml) ± ± ± ±0.4 *lphetil dissimilrities indite signifint differene (p<0.05), **NM = Not mesured Key: DW: distilled wter F: furosemide HCT: hydrohlorothizide SPL: spironoltone UT: urotropin L: lhgi MB: mlt everge IM: intrmusulr; PO: orl Volume: Dt showed tht the highest mount of urine hydrohlorothizide nd orl furosemide (Tle 1), ut volume ws produed fter intrmusulr dministrtion signifint differene (p<0.05) ws only notied etween of furosemide (Tle 1). The differene etween groups the first two groups (lhgi nd mlt everge) nd other reeiving lhgi, mlt everge, hydrohlorothizide, groups. spironoltone nd injeted ontrol groups with the group reeived prenterl furosemide ws signifint Uri id: Birds reeiving prenterl furosemide, orl (p<0.05) with respet to the urine volume. urotropin, hydrohlrothizide, spironoltone nd distilled wter exreted signifintly (p<0.05) higher Gluose: The iggest mount of gluose in urine ws mount of uri id in their urine ompred to other hieved in group reeiving urotropin, prenterl groups (Tle 1). furosemide nd distilled wter. There ws signifint differene (p<0.05) mong orl ontrol with other groups (Tle 1). In ddition, no gluose ws found in the urine smples of orl furosemide, hydrohlorothizide nd Ure nitrogen: lhgi nd mlt everge used signifint (p<0.05) inrese in urinry ure nitrogen ompred to other groups (Tle 1). mlt everge groups. Chloride: The highest levels of hloride ion were exreted in the groups reeiving furosemide (im nd orl). The differene mong these two groups nd other groups ws signifint (p<0.05). lumin: There ws no detetle level of lumin in urine smples of most groups during the period of the experiment exept for the group reeiving hydrohlorothizide. Cretinine: The exretion of retinine ws reltively high fter dministrtion of mlt everge, lhgi, Sodium nd potssium: signifint (p<0.05) nd greter mount of urinry sodium onentrtion ws notied in groups reeiving furosemide nd mlt everge ompred to other groups (Tle 1). similr, ut lower level ws otined for potssium onentrtion in urine. Speifi grvity: Urine speifi grvity ws reltively unhnged (Tle 1). (B) Serum iohemil prmeters: Vlues for vrious serum prmeters fter dministrtion of different ompounds in hens re summrized in Tle

4 Tle 2: Serum levels (Men ± SEM; n = 5) of vrious prmeters 90 minutes fter dministrtion of different gents in lying hens DW (PO) DW (IM) F (IM) F (PO) HCT (PO) Ftors (units) (10 ml/kg) (0.25 ml/kg) (2.5 mg/kg) (5 mg/kg) (10 mg/kg) 226±12 224±8 * 212±11 173±7 212±8 Chloride (meq/l) 108±8 88±17 69±16 120±11 118±8 lumin (g/dl) 2.1± ± ±0.2 NM** 2.1±0.1 Cretinine (mg/dl) 7.1± ± ± ± ±0.2 Uri id (mg/dl) 3.1± ± ± ± ±0.2 Ure nitrogen (mg/dl) 0.6± ± ± ± ± SPL (PO) UT (PO) L (PO) MB (PO) Ftors (units) (1 mg/kg) (50 mg/kg) (10 ml/kg) (10 ml/kg) 204±15 283±32 215±6 229±8 Chloride (meq/l) 116±4 82±15 129±16 113±6 lumin (g/dl) 2.6± ± ± ±0.1 Cretinine (mg/dl) 13.0± ± ± ±0.1 Uri id (mg/dl) 2.1± ± ± ±0.6 Ure nitrogen (mg/dl) 1.7± ± ± ±0.3 *lphetil dissimilrities indite signifint differene (p=0.05), **NM = Not mesured Key: DW; distilled wter F: furosemide HCT: hydrohlorothizide SPL: spironoltone UT: urotropin L: lhgi MB: mlt everge IM: intrmusulr PO: orl Gluose: Serum gluose levels were similr in ll prenterl furosemide nd orl ontrol groups (Tle 2). groups exept for orl furosemide nd urotropin. No signifint differene (p>0.05) in the level of BUN Mximum gluose onentrtion ws found in urotropin ws found mong vrious groups. group. In ontrst, the minimum gluose level ws otined for orl furosemide tril (Tle 2). DISCUSSION Urine olletion: Our ttempt to ollet urine from Chloride: The highest nd the lowest hloride nnulted ureter (Okumur, 1976) ws unsuessful. onentrtion in serum were found in lhgi nd Therefore, urine ws otined y inserting prfinized prenterl furosemide groups, respetively (Tle 2). piee of guze into the lo llowing us to withdrw There ws no signifint (p>0.05) differene etween ll urine y sutioning it from oprodeum using 5 ml other groups. plsti syringe. It is for the first time this method is reported. Employing the method in this experiment for 2 lumin: Mximum nd minimum lumin levels were h used no disomfort to the irds. It should e found in prenterl furosemide nd orl ontrol groups, mentioned tht the urine entering into the oprodeum respetively. No signifint differene (p>0.05) ws should e olleted soon, otherwise it will e prtilly notied mong vrious groups (Tle 2). resored tht my ffet the results. Cretinine: signifintly (p<0.05) greter mount of Norml vlues of serum nd urine iohemil retinine, ws found in ser of irds in group reeiving prmeters in lying hens: Urine nd serum spironoltone. By ontrst, other groups showed non- iohemil prmeters were mesured in irds signifint (p>0.05) lower retinine level. Mximum nd nesthetized with ketmine nd xylzine. So, it is minimum mounts of serum retinine were possile tht the dt reported here e somehow respetively otined for spironoltone nd lhgi different from the rel norml vlues s nesthesi my groups (Tle 2). ffet the onentrtion of these prmeters in serum nd/or urine. Unfortuntely, there is no onsiderle Uri id: lower serum uri id level ws oserved in informtion in this respet nd therefore, the impt of group reeiving prenterl distilled wter ompred to onurrent use of nesthetis on these vlues n not the orl ontrol group. The highest nd the lowest e eluidted or disussed. However, the rnge of mounts of serum uri id were respetively found for iohemil prmeters otined in urine nd serum groups reeiving prenterl furosemide nd distilled re shown in Tle 3. s the dt indites, serum wter. No signifint differene (p>0.05) ws notied gluose vries from mg/dl whih is similr to mong groups. the previous reported vlues (Cmpell nd Coles, 1986; Dein, 1986). Serum lumin onentrtion ws Blood ure nitrogen (BUN): BUN level in urotropin group etween 1.5 nd 3 g/dl whih is lso in greement with ws lose to the orl ontrol group. Mximum nd previous dt reported (Cmpell nd Coles, 1986; minimum BUN levels elonged respetively to the Dein, 1986). Serum retinine level is reported to e 250

5 Tle 3: Norml levels (rnge) of vrious urine nd serum iohemil prmeters in nesthetized lying hens with ketmine nd xylzine Urinry ftors (units) Rnge Serum prmeters (units) Rnge Previous reported vlues* Urine Volume (ml/h) lumin (g/dl) lumin (g/dl) Cretinine (mg/dl) Cretinine (mg/dl) Uri id (mg/dl) Uri id (mg/dl) 5-35 Blood ure nitrogen (mg/dl) Urine ure nitrogen (mg/dl) Chloride (meq/l) Chloride (meq/l) Sodium (meq/l) Potssium (meq/l) Speifi grvity (g/ml) from mg/dl (Cmpell nd Coles, 1986; Dein, 1986) whih is different from the dt otined in the present study (2.50 mg/dl; Tle 3). Serum uri id vlues otined in this study ( mg/dl) lie in the rnge of dt reported elsewhere (25 mg/dl) (Cmpell nd Coles, 1986; Dein, 1986). Serum hloride levels were found to e from 5020 meq/l whih is lose to the previous reported figures (9807 meq/l) (Cmpell nd Coles, 1986; Dein, 1986). No mjor reported dt re ville with respet to the iohemil prmeters in urine. However, the urine speifi grvity found in this experiment ( g/ml) stys within the rnge of g/ml reported y others (Cmpell nd Coles, 1986; Dein, 1986). Diureti effets of dministered gents in hens: Diureti gents my use n inrese in the volume of urine vi vrious mehnisms suh s: ) inhiition of hloride ion resorption followed y positive ions retention in urine (e.g., sodium, potssium nd hydrogen ions) whih onsequently results in higher volume of urine; ) possessing hygrosopi or osmoti hrteristis nd ) inhiition of ntidiureti hormone. The idel diureti gent is the one tht indues the most inrese in urine exretion nd exerts the lest wter nd eletrolyte imlne (Brter, 1998). ltertions in iohemil prmeters, oth in serum nd urine, do not follow regulr pttern in most groups. The most profound hnges re s the following: Urotropin hd reltively high effet on serum gluose in wy tht it used n inrese of out 30% ompred to the ontrol group. Elevtion of gluose sorption from gstro-intestinl trt, inhiition of gluose metolism nd/or inrese in the rte of glyogenolysis my e mong possile mehnisms y whih urotropin exerts its hyperglyemi tion. The other possile mehnism of its tion is the relese of formldehyde in the nephrons whih my ffet the norml struture of glomeruli nd hene exretion of gluose into the urine my our. No onsiderle hnges ourred in serum onentrtion of lumin, retinine, uri id nd BUN. In omprison to the ontrol groups, serum hloride deresed in prenterl furosemide, hydrohlorothizide nd urothropin groups, ut inresed in other groups. Bsed on the proposed mehnism of tion of furosemide in mmmls (Bosh et l., 1977; Hiri et l., 1992), it my inrese hloride ion exretion in urine vi inhiition of its resorption from thik sending lim of Henle s loop nd therefore, exretion of greter - mounts of Cl my derese its serum onentrtion. ording to the dt otined in this study (Tle 1, 2 nd 3), the group reeiving prenterl furosemide produed the highest mount of urine whih is in ordne with the effet in other speies suh s sheep nd mre (Gronwell, 1985; Zimmermn et l., 1978). The urine formed in tht group is round 2-4 times greter thn the urine volume in other groups. The presene of lumin in one urine smple (hydrohlorothizide group; Tle 1) my e due to the ontmintion of urine with diffused mterils (fees or ovidut seretions). The highest nd the lowest mounts of exreted retinine were oserved in groups reeiving prenterl furosemide nd spironoltone, respetively. Prenterl furosemide nd mlt everge were two groups respetively illustrting mximum nd minimum quntities of urine ure nitrogen nd uri id. It ws expeted tht hydrohlorothizide, through ompetition with uri id, uses deline in urinry uri id nd n inrese in serum uri id, ut the dt did not support this phenomenon. Birds in hydrohlorothizide group exreted more hloride in urine possily vi inhiition of sodium resorption in proximl prt of distl tuules. This is supported y derese in serum hloride (Pushett, 1981). No study hs so fr shown the diureti effet of urotropin. s serum hloride ws modertely delined in the group reeiving urotropin ompred to the ontrol groups, it my e dedued this gent n hve wek diureti effet. dose-dependnt inrese in diuresis in ts long with prllel inrese in the exretion of sodium nd hloride ions ws onfirmed y Kltt et l. (1975). In 1976, Cohen nd oworkers proved tht IV dministrtion of furosemide in the dog results in signifint inrese in urine volume, sodium nd potssium. 251

6 Bosh et l. (1977) suggested tht hroni Bosh, J.P., M.H. Goldstein, M.F. Levitt nd T. Khn, dministrtion of furosemide in drinking wter ould enhne urinry sodium exretion in dogs, espeilly in the first dy. Stephens nd Roertson (1985) onfirmed tht ddition of furosemide to the fresh wter inreses urine volume, sodium, potssium nd hloride in turtles whih lk Henle's loop. Vestweer et l. (1989) reported tht IV dministrtion of furosemide in linilly norml Holstein ows rought out sttistil men inrese in urine sodium, potssium nd hloride esides serum sodium. There ws lso signifint derese in the men serum potssium. In 1992, Hiri nd ollegues showed tht IV dministrtion of furosemide uses diureti effet ompnied with n extreme inrese in the exretion rte of sodium nd hloride in the nesthetized dog. Roudeush et l. (1994) me to this onlusion tht furosemide therpy with sodium restrited diet in norml dogs uses no linilly meningful hnges in the serum eletrolytes, ure nitrogen or retinine onentrtions. Spironoltone is elieved to exert its diureti effet through ompetition with ldosterone, so it ws expeted to inrese sodium exretion, ut suh pttern ws not notied in hens. With respet to the hygrophoi nture of urothropin, smll inrese in urine volume plus positive effet on the urinry exretion of sodium, potssium nd hloride n e expeted. Conlusion: dministered drugs nd herl produts hve illustrted different effets on iohemil ftors oth in serum nd urine. Prenterl furosemide ws the most potent diureti drug with respet to the volume of urine produed in lying hens. Furosemide lso exerted the highest influene on the onentrtion of ions inluding sodium, potssium nd hloride, prtiulrly in urine. Similr results were not demonstrted with orl furosemide, even with greter dosge, whih n proly e due to the delyed sorption of the drug from gstrointestinl trt in hens. CKNOWLEGEMENT Finnil support y Shirz University is gretly ppreited. REFERENCES Berg, K.J., ute etylsliyli id poisoning: tretment with fored lkline diuresis nd diuretis. Eur. J. Clin. Phrmol., 12: Brhm, D. nd P. Trinder, n improved olor regent for the determintion of lood gluose y the oxidse system. nlyst J., 97: Brtels, H., M. Bohmer nd C. Heierli, Serum retinine mesurement. Clin. Chem. t, 34: Effet of hroni furosemide dministrtion on hydrogen nd sodium exretion in the dog. m. J. Physiol., 232: Brter, D.C., Diureti Therpy. The New Eng. J. Med., 339: Brun, E.J., Renl funtion. Comp. Biohem. Physiol., 71: Brun, E.J. nd W.H. Dntzler, Funtion of mmmlin-type nd reptilin-type nephrons in kidney of desert quil. m. J. Physiol., 22: Cmpell, T.W. nd E.H. Coles, vin veterinry linil pthology. In: Coles E.H. (Ed) Veterinry Clinil Pthology. Sunders, Phildelphi, pp: Cohen, M.R., E. Hinsh, R. Vergon, J. Ryn, S.J. Kolis nd W.. Shwrtz, omprtive diureti nd tissue distriution study of umetnide nd furosemide in the dog. J. Phrmol. Exp. Therp., 197: Czj, J., H. Moder, W. Wlther, J. Boretius nd K. Wndt, Tretment of ute nd hroni ovine mstitis with injetle Rhodovet, with referene to the linil ourse. Me. Fur. Vet., 38: Dein, F.J., Hemtology. In: Hrrison, G.J., Hrrison, L.R. (Eds). Clinil vin Mediine nd Surgery. Sunders, Phildelphi, pp: Diker, S.E. nd J. Hslm, Wter diuresis in the domesti fowl. J. Physiol., 183: Dormns, T.P., P.G. Gerlg, F.G. Russel nd P. Smiths, Comintion diureti therpy in severe ongestive hert filure. Drugs, 55: Doums, B.T., W.. Wtson nd H.G. Biggs, lumin stndrds nd the mesurement of serum lumin with romresol green. Clin. Chem. t, 31: Emery, N., T.L. Poulson nd W.B. Kinter, Prodution of onentrted urine y vin kidneys. m. J. Physiol., 223: Evns, R.T., Mnul nd utomted methods for mesuring ure sed on modifition of its retion with dietyl monoxime nd the semirzide. J. Clin. Pthol., 21: Finy, D.L. nd G. Ertingshusen, utomted retion-rte method for determintion of serum retinine with the Centrifihem. Clin. Chem., 17: 391. Gronwell, R., Effet of diuresis on urinry exretion nd retinine lerne in the horse. m. J. Vet. Res., 46: Hiri, J., H. Miyzki nd T. Tneike, The phrmokinetis nd phrmodynmis of furosemide in the nesthetized dog. J. Vet. Phrmol. Therp., 15:

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