Effect of different infective doses of Ascaridia galli eggs on the total serum protein and weight gain in white lohman laying hens

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1 Effect of different infective doses of scridi glli eggs on the totl serum protein nd weight gin in white lohmn lying hens * mer.m. l-mery * Thury.kh. l-drji *Deprtment of Prsitology-Veterinry Medicine College-ghdd University-ghdd Irq. m.murhum@yhoo.com bstrct The present study ws conducted to investigte the effect of different infective doses of scridi glli eggs on the totl serum protein, weight gin nd shedding of eggs per grm (EPG) in lying hens by using 75 lyer birds, eight weeks old which divided rndomly into 3 groups, ech group included 25 chickens.1st nd 2 nd groups were infected orlly with 1500 nd 250 vible. glli eggs/ bird respectively. The 3 rd group ws given 0.5 ml physiologicl sline orlly (control negtive). The results of the totl serum protein concentrtion decresed significntly in both infected (1 st nd 2 nd groups) s compred with the control non infected group nd ws decresed significntly in the first infected group thn the second infected group. Control group ws showed higher (p<0.05) of weights thn the first nd second infected groups,while the first group recorded significnt decrese (p<0.05) thn the second group. The number of eggs count (EPG) in feces of infected birds ws higher in the first thn in the second group. KEY WORD: lying hens, scridi glli, serum protein, weight gin,ghdd. تا ثیر الا صابة بجرع مختلفه ببیوض على scridi طفیلي glli تركیز البروتین معدل الكليو الدجاج في الاوزان اللوهمان الابیض * ثریا خالد عبدالواحد الدراجي * عامر مرحم العامري * فرع الطفیلیات / كلیة الطب البیطري /جامعة بغداد /العراق. 1

2 الخلاصة هد فت الدراسة الحالیة ا لتحر ي عن تا ثیرات الاصابة التجریبیه بطفیلي scridi glli في الدجاج البیاض من ٣ نوع لوهمان الابیض با ستعمال ٧٥ دجاجة بیاضة بعمر ثمانیة أسابیع والتي قس مت بشكل عشواي ي إلى مجامیع تضم نت كل مجموعة ٢٥ دجاجة لمده ٨ اسابیع. المجموعة الا ولى والثانیة أصیبت ب ١٥٠٠ بیضة مصیبة / طیر عن طریق الفم و ٢٥٠ بیضة عیوشه / طیر عن طریق الفم على التوالي أما المجموعة الثالثة (سیطرة سالبة) أعطت المحلول الملحي الفسلجي عن طریق الفم كمجموعة سیطرة.كما أظهرت النتاي ج الى وجود أنخفاضمعنو ي( 0.05> p ( الكلي البروتین تركیز معدل في في المجموعتین المصابتین الاولى والثانیة مقارنة بمجموعة السیطرة في الشهر الاول واستمر في الانخفاض الشهر الثاني.أظهرت مجموعة السیطرة معدل اوزان اعلى مقارنة مع المجموعتین الاولى والثانیة وسجلت المجموعة الاولى انخفاضا اكثر من المجموعة الثانیة. في حین كانت اعداد بیوض الطفیلي لكل غرام من براز الطیور المصابة اعلى في المجموعة الاولى مقارنة مع المجموعة الثانیة. Introduction scridi glli is n intestinl helminth prsite belonging to the group of scrid worms (Phylum Nemtod; Clss Secernenti; Order scridid; Fmily scridiide). Members of the genus scridi re essentilly prsitic in birds, nd most prevlent in fowl, prticulrly in chicken nd turkey, geese, guine fowl nd number of wild birds; the principl host mnifestly being the chicken. (1) Studies hve suggested tht. glli is the most common nemtode in ll types of production systems nd hs worldwide distribution. (2). Infections with scridi spp. my cuse, in ddition to direct losses, reductions in growth rte, weight loss, reduced feed conversion rtes nd dmge to the intestinl mucos, leding to blood loss nd secondry infections (3). prtil or complete obstruction of the intestine, nd incresed mortlity due to secondry bcteril infections.(4). The primry dmge reduced efficiency of feed utiliztion, reduced egg production nd weight loss re common symptoms in broiler chickens; Young birds re most susceptible, nd hevier breeds seem more resistnt thn the lighter breeds such s leghorns nd white minorcs (5). The life cycle of. glli is direct nd the infective stge is eggs hrbouring 2

3 third stge lrve. fter ingestion, eggs rech the duodenum nd htch within 24 hours (6), nd lrve re relesed into the lumen of the intestine. The lrve then enter the mucos of the smll intestine nd most lrve initite histotrophic phse between dy 8 to 17 post infection (p.i.) (7).. glli my lso ply role in trnsmission of Slmonell infections (8) nd vin Reo viruses (9) resulting in disese nd economic loses. This study imed to study effect of. glli infection on in lyer hens using different doses on the totl serum protein nd weight gin. Mterils nd Methods 1- Preprtion of scridi glli eggs: dult scridi glli femle worms were collected from nturlly infected chickens ccording to stndrd prsitologicl techniques ccordig to (10,3). The uteri of grved femle worms were dissected nd the eggs were recovered. The eggs embryonted by incubtion in 0.1% (w/v) potssium-dichromte solution t room temperture for 14 dys(11). Lrve vibility ws ssessed by observing the lrve spontneous movement inside the egg fter incresing the surrounding temperture. Infective. glli L3 eggs were stored t +4 C until use. On the dy of infection nother vibility test ws performed. Only lrve tht were well developed, motile nd not htched were ccounted s vible nd infective. Eggs in the culture were suspended with tp wter to get finl volume of 0.5 ml contining the infection dose to be given to ech bird. Number of eggs/ml suspension ws determined using McMster egg counting chmber. The chickens inoculted vi orl gvge t 8 weeks old with n infection doses of 1500 nd 250 embryonted eggs per bird. 2- Experimentl Design:- Seventy five chickens (8 weeks old ) white lohmn lying hens were purchsed from locl lyer field nd kept in room 25 m 2 of n experimentl niml house - Veterinry Medicine College - ghdd University,which hd free ccess to wter, nd the diet ws good grounded seeds with supplements vitmins nd mino cids with nticoccidil drugs. (12). Chickens were divided rndomly into three groups (25 chicks ech) ; The hens were orlly infected using plstic Psteur pipette s described by (10); The 1 st nd 2 nd groups 3

4 were inoculted with 1500 nd 250 infected egg / bird respectively,3 rd group were inoculted with 0.1 ml of phosphte buffer sline( PS) orlly s negtive control group. 3-Estimtion of totl serum protein concentrtion: fter collection of blood from the hns through wing vein, blood ws trnsferred into gel tube (without nticogulnt) nd llowed to stnd in refrigertor for 15 minutes nd centrifuged t speed of up to 3500 RPM for 5 minutes. Serum ws spirted by pipette nd trnsferred into smll clen tubes kept frozen until use. Colorimetriclly totl serum protein mg/dl ws estimted by the iuret method s discribed by (13) using SPN dignostic kit (Code N ). 4- ody weight:- Five chicks were rndomly weighed of ech group weekly for 7 intervls time (14). 5- Egg counts - Eggs per Grm of feces (EPG): The eggs were counted using modified McMster method (15). 6-Sttisticl nlysis:- Sttisticl nlysis of mens were performed by using sttisticl pckge for socil science (SPSS,2008), Version 16, nd for determintion of significnt differences by using two wy nlysis NOV (16). Results nd discussion 1-Totl protein mg /ml: Tble(1) shows significnt decrese (P<0.05) in totl protien between the 1 st nd 2 nd groups tht infected with 1500 nd 250 vible eggs t the 3 rd nd 4 th weeks respectively lso there is significnt decrese (P<0.05) within group in the 1 st nd 2 nd groups t the 2 nd, 3 rd nd 4 th week. In the second month between groups; significnt decrese (P<0.05) ws found between the 1 st nd control groups t 6 th,7 th nd 8 th weeks, but only significnt decrese (P<0.05) ws recorded between the second nd control groups t 7 th week. (first nd second groups) showed no significnt differences (P>0.05)in compre with control birds. (Tble 2). 4

5 Tble 1: Effect of.glli infection on the Totl proteins mg/ml in lying chickens in the First month: Weeks Groups Group ± 0.31 Men±SE (%) First month ± 0.18 b 2.64 ± 0.18 C b 2.38 ± 0.34 b Group ± ± 0.18 b 3.63 ± 0.37 b 3.62 ± 0.31 b Group ± ± ± ± 0.37 Different cpitl letters show significnt difference (P<0.05) between groups. Different smll letters show significnt difference (P<0.05)within group. SE= Stndrd error. N= 5 nimls ech group.g1= infected with (1500 vible egg / bird) G2= infected with (250 vible egg /bird) G3= Control negtive. Tble 2: Effect of.glli infection in Totl proteins mg/ml in lying hens in the secod month: Men±SE (%) Weeks Groups Second month Group ± ± ± ± 0.27 C Group ± ± ± ± 0.36 Group ± ± ± ± 0.42 Different cpitl letters show significnt difference (P<0.05) between groups. Different smll letters show significnt difference (P<0.05)within group. SE= Stndrd error. N= 5 nimls ech group.g1= infected with (1500 vible egg / bird) G2= infected with (250 vible egg /bird) G3= Control negtive. 5

6 iochemicl study showed tht significntly decresed in totl serum protein in both infected groups with scrdi glli s compred with control group. Our finding ws in greed with tht of (17). Coles (18) reported tht considerble loss of digestive secretion nd mucous due to intestinl prsitism in nemic birds, which lso cuse inefficient protein bsorption nd utiliztion in the system to extent of leding to mrked decrese in serum protein The lowered of totl protein vlues my result from gret loss of tissue protein occuring through lekge into gut with loss of digestive secretion nd mucous due to intestinl prsitism in nemic birds,which lso cused inefficient protein bsorption nd utiliztion process (19). Similr phenomen my occur in chicken following.glli infection in this study. 2. ody weight : In the first month; there ws significnt decrese (P<0.05) between the 1 st nd the control groups t 3 rd nd 4 th week, while no significnt difference (p>0.05) ws found between the 1 st nd 2 nd group nd between the 2 nd nd control group. In the second month there ws significnt decrese (P<0.05) between the 1 st group nd 2 nd group compred with the 3 rd group. Tble 3: Effect of.glli infection on body gins (gm/bird) in lying hens the first month. Different cpitl letters show significnt difference (P<0.05) between groups. SE= Stndrd error. N= 5 nimls ech Men±SE (%) Weeks Groups First month Group 1 710± ± ± ±33.7 Group 2 720± ± ± ±30.5 Group 3 690± ± ± ±43.1 6

7 group.g1= infected with (1500 vible egg / bird) G2= infected with (250 vible egg /bird) G3= Control negtive. Tble 4: Effect of.glli infection on body gins (gm/bird) in lying hens hens in the second month: Weeks Groups Men±SE (%) secondmonth Group ±18.7 C 1130±19.7 C 1244 ± C 1272± 21.3 C Group ± ± ± ±14.2 Group ± ± ± ±52.2 Different cpitl letters show significnt difference (P<0.05) between groups. SE= Stndrd error. N= 5 nimls ech group.g1= infected with (1500 vible egg / bird) G2= infected with (250 vible egg /bird) G3= Control negtive. The development of immunity to prsitic infections is costly process nd requires high metbolic inputs (20). Mrcos-txutegi (21) hve shown tht infected chickens produce incresing mounts of specific IgG ntibodies ginst both embryonted. glli egg ntigens nd dult somtic ntigens from the first week of infection to dy 42. In this period, the ctivity of histotropic stges of the prsite nd host immunity re developing, the histotropic phse is norml prt of the life cycle of. glli nd it lsts pproximtely 7 50 dys, depending on number of prsite nd host-oriented fctors (7). lso, the pthogenicity of. glli is considered to be stronger during histotropic, lrvl development, resulting in inflmmtion nd injury to the intestinl wll nd to the host s bsorption of metbolic wste (22). Correspondingly, the lloction of nutrients my primrily be chnged from growth to cquisition of immunity when growing niml first encounters prsitic (tht my be ssumed tht the lower body weight development of infected compred to uninfected birds (23).. glli infection my lso influence digestion nd bsorption of nutrients by reducing proteolytic enzyme ctivity in the jejunum (24) being ssocited with n incresed recycling of nitrogen (25). Reduced metbolizbility of energy 7

8 nd lowered nitrogen retention hve been reported for. glli infected chickens (26). Recent dt suggest tht. glli infection impirs electrogenic trnsport of lnin nd glucose in the intestinl epithelil tissues of growing chicks (27). 3. Shedding of eggs: The mens of egg per grm in the first group (received 1500 vible eggs)ws higher thn the second group (infected with 250 vible eggs )with significnt increse (P<0.05) t the 5 th week till the end of the experiment. ( Tble 5) Tble 5:Number of egg per grm (EPG) in litter of infected groups with.glli infection in lying hens Weeks Groups Group ± ± ± ± Group ± ± ± ± N= 5 nimls ech group.g1= infected with (1500 vible egg / bird) G2= infected with (250 vibl e egg /bird). The number of EPG my be influenced by the number of dult worms in the gstrointestinl trct, worm ge, host immunity, host ge, host sex, stges of infection, fecundity, feed composition nd consistency of the feces nd time of dy of collecting the feces,even though the EPG cnnot be used directly to estimte the worm burden in either experimentl infections or in nturl infections, the described method proved dequte for our experiments, s the sensitivity ws high mking it possible for us to determine when the worms becme mture (28) 8

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