Haematological and Biochemical Changes in Japanese Quails Coturnix coturnix Japonica and Chickens Due to Ascaridia galli Infection

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1 Interntionl Journl of Poultry Science 7 (7): , 2008 ISSN Asin Network for Scientific Informtion, 2008 Hemtologicl nd Biochemicl Chnges in Jpnese Quils Coturnix coturnix Jponic nd Chickens Due to Ascridi glli Infection 2 K. Dek nd J. Borh Leoprd Rescue Centre, Junnr, Mhrstr, Indi Wildlife Institute of Indi, Post Box 8, Chndrbni, Dehrdun-24800, Uttrkhnd, Indi 2 Abstrct: A study ws crried out in Jpnese quils Coturnix coturnix jponic nd chickens (Rhode Islnd Red) for comprtive hemtologicl nd biochemicl chnges occurring due to Ascridi glli infection. Quils s well s chickens were divided into four smll groups, ech consisting six numbers of birds in ech group nd were mrked s q, q2, q3, qc nd p, p2, p3, pc, respectively. Hemtologicl study showed tht the totl erythrocytic count (TEC), pcked cell volume (PCV) nd hemoglobin (Hb) percentge decresed significntly in infected groups of quils nd chickens. The totl leucocytic count (TLC) showed significnt increse in infected groups of quils nd chickens. Heterophils nd eosinophils were incresed significntly in ll the infected groups of quils nd chickens. Lymphocytes decresed significntly in ll the infected groups. Biochemicl study showed tht totl serum protein decresed significntly in the infected groups of quils nd chickens. Serum lbumin level ws significntly lower in ll the infected groups of quils nd chickens. Serum globulin nd lbumin: globulin (A:G) rtio filed to show ny significnt difference between control nd infected groups of quils nd chickens. Key words: Jpnese quils, chickens, hemtologicl chnges, biochemicl chnges, Ascrdi glli Introduction In most of the developing countries, including Indi, teeming with millions of poultry, re t stge of perpetul protein hunger. Poultry met nd eggs, though the mjor source of niml protein, is still now unble to meet up the protein hunger of the world (Gffer, 986). During the lst two decdes, Indi hs hd remrkble growth in poultry industry. Indi s egg production ws 2 million tones in the yer 2002 nd remined mongst the top 5 of egg producing countries in the world. The broiler met production ws.566 million tones in the sme yer (Poultry Interntionl, Vol- 42, No., Nov. 2003). Nowdys, quil frming is cropping up s new venture of diversifiction of poultry frming, not only to diverse the choice of tste but lso to strengthen the met production unit for fulfilling the shortge of niml protein demnds mongst the Non-Veg peoples of Indi. In Indi, Jpnese quil (Coturnix coturnix jponic) breeding for egg nd met production hs been introduced very recently. Since 974, reserch work on the vrious spects of quil rering such s breeding, feeding nd mngement, disese control etc. hve been tken up by the Centrl Avin Reserch Institute of Iztngr nd it is ttrcting the poultry frmers nd consumers for quils egg nd met. Quils nd domesticted fowls belong to the sme subfmily Phsioxids. Quils possess n excellent disese resistnce qulity thn those of chickens nd hve been chosen for its economicl vibility in frming. Jpnese quils re rpidly gining populrity for its commercil exploittion nd in ner future, my cquire n importnt segment in rpidly expnding Indin poultry industry. Among the helminth prsites, Ascridi glli is most prevlent species occurring in most of the fowls. Due to the close ssocition nd frequent indirect contct between the quil nd chicken rering units in the sme premises, there is possibility tht Ascridi glli might estblish themselves s prsite for the quils s new host (Singh et l., 996). It will not only directly cuse dmges to the host but lso my predispose the quils to other infections. The present experiment ws crried out to observe comprtive hemtologicl nd biochemicl chnges in Jpnese quils nd chickens tht occurs due to Ascridi glli infection. Mterils nd Methods Source: A totl of 24 dy old chicks of Jpnese quil nd sme number of Rhode Islnd Red (RIR) dy old chicks were obtined from Stte Poultry Frm, Tollygunj, Kolkt, Indi. Feeding nd mngement: The Jpnese quil chicks nd poultry chicks were rered seprtely in brooder t experimentl house for two weeks. Afterwrds, the poultry nd quil chicks were rndomly distributed in four groups of ech with three infected nd one control group s PC, p, p2 nd p3 nd Q C, q, q2 nd q3 respectively. Ech smll group contined six numbers of birds nd ws mintined in seprte wire mesh bttery 704

2 Dek nd Borh: Jpnese Quils cges. Ech bird ws mrked with leg bnds. Regulr clening nd septic mesures were followed with regulr feeding nd wtering. The quils were mintined on blnced broiler strter rtion throughout the life. The fowl chicks were lso offered broiler strter rtion up to three weeks of ge nd then ws shifted to blnced poultry rtion till the end of the experiment. Collection nd culture of Ascridi glli eggs: The dult femle worms of Ascridi glli were collected from the intestine of desi (locl) fowl, procured from poultry slughter houses. The worms were thoroughly wshed in norml sline solution to remove the mucous nd other debris s. The nterior portion of the worm ws tken on slide nd ws cut off ner the vgin by shrp sclpel. The worm ws then held t the posterior end by forceps nd pressure ws pplied by needle to tkeout the grvid uterus contining mture eggs. The uterus ws then disintegrted with the help of needle to get the eggs free from the uterus. It ws then cultured in 0.5% formlin solution in petridish to prevent bcteril growth. The culture ws shken dily for ertion. Fresh medi ws dded to replenish the volume. The development of eggs ws exmined regulrly till they rech the infective stge. The second stge infective lrve developed in 5-20 dys in the B.O.D. incubtor. The temperture in B.O.D. ws mintined t 28± C s described by Mllik (98). Counting of infective eggs: When infective stge ws reched, the eggs were collected in centrifuge tube contining sline nd centrifuged t 000 rpm for five minutes, fter which the superntnt fluid ws discrded. The process ws repeted three times. Then smll quntity of distilled wter ws dded to the sediment nd mixed thoroughly. Finlly the egg suspension ws prepred in mesured volume of distilled wter. From this, 0. ml of homogenous suspension ws tken on microslide nd covered with coverslip. The numbers of infective Ascridi glli egg in 0. ml of the suspension ws counted under compound microscope. Five such counts were mde nd the men infective egg concentrtion per 0. ml of suspension ws clculted (Choudhury, 989). The infective doses of eggs i.e., 00, 500 nd 000 numbers of eggs were djusted in, nd 2 ml of the suspensions respectively. Experimentl designs: Chickens s well s quils were divided into four smll groups with ech consisting 6 number of birds in ech group. Chicken groups were mrked s pc, p, p2, p3 nd quil groups were mrked s qc, q, q2 nd q3. Group pc nd qc: The birds of these two groups were kept s control for comprison of ll the prmeters with infected groups. Group p nd q: Ech bird of these groups ws infected orlly with one hundred infective eggs t the ge of twenty one dys. Food nd wter ws withdrwn for twelve hours before dministrtion of infection (Shilskr nd Prsr, 985). After giving infection birds were kept under observtion to see the effect of infection. Group p2 nd q2: Ech bird of these groups ws infected orlly with five hundred infective eggs on twenty one dys ge. The procedure dopted for giving infection ws sme s Group q nd p. Group p3 nd q3: Ech bird of these groups ws lso infected orlly with one thousnd infective eggs t the sme ge nd the procedure of giving infection ws sme s previous groups. All the birds were scrificed on 9th week post infection. Methods Hemtologicl: Blood ws collected from the individul birds of ech group t the time of slughter from jugulr vein. Sterile vils with 20 µl of 0% EDTA were used s nticogulnt for collection of blood. Two milliliters of nti-cogulted blood ws collected from ech bird nd ws kept in refrigertor for hemtologicl studies. TEC nd TLC were done by Neubuer hemocytometer. The Rees nd Ecker solution ws used s diluting fluid s described by Sstry (983). DLC ws estimted by using Wright-Giems stin s per method described by Schlm et l. (986). Hb concentrtion ws estimted by cynmethemoglobin method s described by Dcil (985). PCV ws determined by Wintrobe hemtocrit method s described by Schlm et l. (986). Biochemicl: Blood ws collected from the individul birds of ech group t the time of slughter from jugulr vein. Two milliliters of blood ws collected from ech bird in sterile test tubes without nticogulnt nd llowed to clot. Serum ws seprted out nd kept t 20 C until nlysis. Totl protein (g dlg ) nd lbumin (g dlg ) were estimted by Biuret nd Dums method s described by Dums et l. (97) by using SPAN dignostic kit (Code No. 2593). Serum globulin (g dlg ) ws estimted s difference between totl protein nd lbumin. The A:G rtio ws clculted by dividing the concentrtion of lbumin in (g dlg ) by concentrtion globulin (g dlg ). Sttisticl nlysis: All the dt obtined in respect to hemtologicl nd biochemicl prmeters studied during experiment were sttisticlly nlysed for test of significnt (Tble 3-6) s per sttisticl methods of Snedecor nd Cochrn (967). 705

3 Dek nd Borh: Jpnese Quils Results nd Discussion Hemtologicl chnges: In quils, TEC decresed significntly (p<0.0) in group q nd q2, but group q3 were non significnt with tht of control group. Tnwr et l. (200) lso found similr observtion in their experiment. TEC vlues were significntly lower by % level in ll the infected groups of chicken thn tht of control group. Lowered TEC in Ascridi glli infected chickens nd quils might be due to lowered erythropoesis. A. glli re usully ssocited with mild/cute enteritis which hmpers the bsorption of essentil nutrients for blood cell formtion. Group q3 showed no significnt chnge in TEC. In quils, PCV decresed significntly (p<0.05) in ll the infected groups thn tht of control group. PCV percentge in chickens of group p nd p2 decresed significntly (p<0.0) nd in group p3 the sme ws decresed (p<0.05) to tht of control group. Mtt nd Ahluwli (982) recorded the sme finding in their experiment with fowls infected with Ascridi glli. PCV my hve decresed due to the lower concentrtion of erythrocytes per unit volume of blood in the infected group of chickens nd quils. The hemoglobin percentge lowered significntly (p<0.05) in ll the infected groups of chicken in comprison to tht of control group. In quil, Hb percentge reduced in group q nd q2 (p<0.0) nd in group q3 (p<0.05) to tht of control group. This finding is in concordnce with the finding of Mtt nd Ahluwli (982) nd Kumr et l. (2003). They opined tht lowered hemoglobin concentrtion in infected birds ws correlted with the ctivities of erly lrvl stge of A. glli in the process of penetrtion with resultnt destruction of mucos of smll intestine nd rupture of smll blood vessels. Kumr et l. (2003) lso cited tht fll of Hb content might be due to metbolic disturbnce cused by worms rther thn direct blood loss. In the present study, the TLC in quils were significntly (p<0.0) higher in ll the infected groups to tht of control group. The TLC in chickens of group p nd p2 incresed significntly (p<0.05) wheres group p3 did not show significnt increse to tht of control group. This is in greement with the findings of Tnwr nd Mishr (200). In chickens, the heterophil percentge incresed significntly (p<0.0) in group p nd p2 while in group p3 it ws incresed (p<0.05) to tht of control group. The quils of group q, q2 nd q3 lso showed the similr results with tht of chicken nd this finding simulted the findings of Tnwr nd Mishr (200). They expressed tht heterophils re ctively moeboid nd phgocytic in Tble : Hemtologicl profiles of Jpnese quils nd chicke infected with scridi glli TEC Hb PCV TLC Heterophil Eosinophil bsophil Lymphocytes Monocytes Groups 6 (0 /Cu mm) (%) (%) 3 (0 /Cu mm) (%) (%) (%) (%) (%) qc ±0.20 ±0.308 ±.235 ±0.664 ±0.558 ±0.428 ±0.224 ±0.577 ±0.422 pc ±0.59 ±0.608 ±.349 ±0.966 ±0.764 ±0.477 ±0.40 ±0.65 ±0.543 q ±0.25 ±0.480 ±0.25 ±0.763 ±.67 ±0.495 ±0.2 ±0.946 ±0.823 p *c ±0.206 ±0.630 ±0.846 ±0.725 ±0.730 ±0.847 ±0.667 ±.230 ±0.477 q ±0.75 ±0.324 ±0.75 ±0.527 ±.470 ±0.633 ±0.40 ±.283 ±0.75 p ±0.02 ±0.602 ±0.994 ±0.629 ±.7 ±0.847 ±0.495 ±.887 ±0.882 q ±0.3 ±0.335 ±0.335 ±0.53 ±.302 ±0.577 ±0.333 ±.336 ±0.365 p ±0.50 ±0.68 ±0.85 ±0.68 ±0.667 ±0.60 ±0.422 ±.797 ±0.428 **Men±SE (row-wise) bering different superscripts differs significntly (p<0.0); *Men±SE (row-wise) bering different superscripts differs significntly (p<0.05); Non significnt Tble 2: Bio-chemicl profiles of Jpnese quils nd chickens infected with Ascridi glli [Men±SE] Groups Totl protein (gm/00 ml) lbumin (gm/00 ml) Globulin (gm/00 ml) A:G rtio qc ± ± ± pc ± ± ± ±0.24 q ± ± ± ±0.038 p ± ± ± ±0.043 q ± ± ± ±0.054 p ± ± ± ±0.067 q ± ± ± ±0.066 p ± ± ± ±0.072 **Men±SE (row-wise) bering different superscripts differs significntly (p<0.0); *Men±SE (row-wise) bering different superscripts differs significntly (p<0.05); :Non significnt 706

4 Dek nd Borh: Jpnese Quils Fig. : Hemtologicl profiles of the chickens infected with A. glli Fig. 2: Hemtologicl profiles of the Jpnese quils infected with A. glli nture. The phgocytic ction of heterophils my thus observed tht incresed number of eosinophils in the correlte with their incresed number s first line of blood is n indiction of prsitic infection. Tnwr nd defence of the host in the present study. Eosinophil Mishr (200) reported tht the net increse in the totl percentge in chicken incresed significntly (p<0.0) in leucocytic count might be due to the increse in group p nd p2 while in group p3 the sme ws heterophils nd eosinophils. incresed to p<0.05. In quils the significnt increse of In the present study, the lymphocyte percentge were eosinophils in ll the infected groups ws (P<0.0) in significntly decresed (p<0.0) in ll the infected comprison to tht of control group. groups of chickens nd quils to tht of control group. Tnwr nd Mishr (200) nd Kumr et l. (2003) hd This finding ws in ccord with the findings of Tnwr lso observed the sme result in their experiment. They nd Mishr (200). 707

5 Dek nd Borh: Jpnese Quils Tble 3: Anlysis of vrince of hemtologicl prmeters in chickens Tble 4: Anlysis of vrince of hemtologicl prmeters in quils infected with Ascridi glli infected with Ascridi glli 6 6 TEC (0 /Cu mm) TEC (0 /Cu mm) Between groups ** Between groups ** Within groups (Error) Within groups (Error) Hb (gm/dl) Hb (gm/dl) Between groups * Between groups ** Within groups (Error) Within groups (Error) PCV (%) PCV (%) Between groups ** Between groups ** Within groups (Error) Within groups (Error) TLC (0 /Cu mm) TLC (0 /Cu mm) Between groups * Between groups ** Within groups (Error) Within groups (Error) Heterophil (%) Heterophil (%) Between groups ** Between groups ** Within groups (Error) Within groups (Error) Eosinophil (%) Eosinophil (%) Between groups ** Between groups ** Within groups (Error) Within groups (Error) Bsophil (%) Bsophil (%) Between groups Between groups Within groups (Error) Within groups (Error) Lymphocyte (%) Lymphocyte (%) Between groups ** Between groups ** Within groups (Error) Within groups (Error) Monocyte (%) Monocyte (%) Between groups * Between groups Within groups (Error) Within groups (Error) **(p<0.0), *(p<0.05), :Non significnt **(p<0.0), *(p<0.05), :Non significnt Chickens of group p showed significnt (p<0.0) increse in monocyte percentge wheres group p2 nd p3 were non significnt to tht of control group. In ll the infected groups of quil, the percentge of monocytes were non significnt to tht of control group. The dt s re summrized in Tble. Biochemicl chnges: In the present study, the totl protein showed significnt decrese (p<0.05) in group q nd q2 while group q3 filed to show ny significnt difference with control group. In chickens, the totl protein ws significntly lower in group p nd p2 (p<0.0) rther thn in group p3 (p<0.05). This finding ws in greement with the findings of Tnwr nd Mishr (200). Hypoprotenemi might occur due to incresed motility of intestine s in dirrhoe. In tht cse the proteins might get lost from the bowel. Coles (967) reported tht considerble loss of tissue protein my occur through lekge into gut with 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 the extent of leding to mrked decrese in serum protein. In quils, the entire three infected group showed significnt (p<0.0) lower lbumin level thn tht of the control group. The level of lbumin ws significntly lower in chickens of group p nd p2 (p<0.0) while in group p3 the sme ws decresed (p<0.05). The decrese in lbumin concentrtion is common 708

6 Dek nd Borh: Jpnese Quils Tble 5: Anlysis vrince of bio-chemicl prmeters in chickens infected with Ascridi glli Totl protein (gm/00 ml) Albumin (gm/00 ml) Globulin (gm/00 ml) A : G rtio SOV df SS MS F SS MS F SS MS F SS MS F Between groups ** ** Within groups (Error) **(p<0.0), :Non significnt Tble 6: Anlysis vrince of bio-chemicl prmeters in quils infected with Ascridi glli Totl protein (gm/00 ml) Albumin (gm/00 ml) Globulin (gm/00 ml) A : G rtio SOV df SS MS F SS MS F SS MS F SS MS F Between groups * ** Within groups (Error) **(p<0.0), *(p<0.05), :Non significnt. SOV: Source of vrition. df: degree of freedom Fig. 3: Fig 4: Biochemicl profiles of chickens infected with A. glli Biochemicl profile of Jpnese quils infected with A. glli Conclusion: Hemtologicl study showed tht the totl erythrocytic count (TEC), pcked cell volume (PCV) nd hemoglobin (Hb) percentge decresed significntly in infected groups of quils nd chickens except tht group q3, which did not show ny significnt difference in TEC nd PCV when compred to the control group. The totl leucocytic count (TLC) showed significnt increse in infected groups of quils nd chickens wheres group p3 filed to show significnt difference. Heterophils nd eosinophils incresed significntly in ll the infected groups of quils nd chickens wheres monocytes incresed only in group p. Lymphocytes decresed significntly in ll the infected groups. Biochemicl studies showed tht totl serum protein decresed significntly in the infected groups of quils nd chickens except the group q3, which ws non significnt. Serum lbumin level ws significntly lower in ll the infected groups of quils nd chickens. Serum globulin nd lbumin:globulin (A:G) rtio filed to show ny significnt difference between control nd infected groups of quils nd chickens. From the bove study, it ws observed tht quils were lso susceptible to helminth prsites which re usully prevlent in chicken, but the quils re slightly more resistnt to infections thn chickens. As the quils possess n excellent disese resistnce qulity ginst the prsitic infection thn those of chickens, they cn be chosen for economicl vibility in frming. form of hypoproteinemi due to its smll size nd osmotic sensitivity to fluid movement. The lbumin is selectively lost in intestinl prsitism. The hypolbuminemi of intestinl prsitism is ggrvted by incresed lbumin ctbolism (Tnwr nd Mishr, 200). Globulin percentge in the infected groups of chicken nd quil did not show significnt chnges with tht of control groups nd re in conformity with the findings of Tnwr nd Mishr (200). In the present study, the protein profile of both chicken nd quils indicted hypoproteinemi with no chnge in globulin nd in A:G rtio. The results re summrized in Tble 2. Acknowledgement The uthors re thnkful to Dr. C. K. Dsgupt nd Dr. M. C. Bndopdhyy, Professors, Deprtment of Veterinry Prsitology, West Bengl University of Animl nd Fishery Science, Kolkt, Indi, for their kind help to crry out this study. References Choudhury, S., 989. Studies on experimentl Heterkis gllinrum infection in chicken. M.V.Sc. Thesis submitted to Assm Agriculturl University, pp: Coles, E.H., 967. Veterinry Clinicl Pthology. 2nd Edn. W. B. Sunders Co., Phildelphi. 709

7 Dek nd Borh: Jpnese Quils Dcil, J.V., 985. Prcticl Hemtology. 6th Edn. Dums, B.T., R.L. Arends nd P.V.C. Pinto, 97. Determintion of serum lbumin using BCG. In: Stndrd Methods Clin. Chem., 7 : Gffer, T.A., 986. Still fr, fr to go. Poult. Adviser, XIX: 5-6. Kumr, R., S.R.P. Sinh nd S.B. Verm, Hemtologicl chnges in Jpnese quils nturlly infected with Rillietin tetrgon. J. Vet. Prsitol., 6: Kumr, R., S.R.P. Sinh, S.B. Verm nd S. Sinh, Hemtologicl chnges in the Jpnese quils (Coturnix coturnix jponic) nturlly infected with nemtode Ascridi glli. Ind. Vet. Med. J., 27: Kumr, R., S.R.P. Sinh, S.B. Verm nd S. Sinh, Hemtologicl chnges in the Jpnese quils (Coturnix coturnix jponic) nturlly infected with nemtode Ascridi glli. Ind. Vet. Med. J., 27: Mllik, A.K., 98. Studies on the round worms of poultry with specil reference to immunologicl response to Ascridi glli infection. M.V.Sc. Thesis submitted to Bidhn Chndr Krishi Viswvidyly, pp: 45. Mtt, S.C. nd S.S. Ahluwli, 982. Hemtologicl indices s influenced by Ascridi glli infection in fowl. Effect on the hemoglobin concentrtion, pcked cell volume nd erythrocytes sedimenttion rte. Ind. J. Poult. Sci., 7: Sstry, G.A., 983. Veterinry Pthology. 6th Edn. CBS Publishers nd Distributors. New Delhi-0 032, pp: 727. Schlm, O.W., N.C. Jin nd E.J. Corroll, 986. Veterinry Hemtology. 4th Edn. Le nd Febiger, Phildelphi. Shilskr, D.V. nd G.C. Prsr, 985. In vivo nd Kymogrphic studies on Psorle corylifoli nd Piper betle ginst vin Ascridi glli. Ind. Vet. J., 62: Singh, B., M.S. Oberoi, S.K. Jnd nd A. Singh, 996. Emerging diseses of poultry in Indi. J. Res. Punjb Agric. Univ., 33: Snedecor, G.O. nd W.G. Cochrn, 967. Sttisticl methods. Oxford nd IBH Publ. Co., Jnpth, New Delhi. Tnwr, R.K. nd S. Mishr, 200. Clinico-Hemto- Biochemicl studies on intestinl helminthisis in poultry. Vet. Prctitioner, 2: