B.A. Usman, A.U. Mani, A.D. El - Yuguda and S.S. Diarra 1

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1 Internatinal Jurnal f Pultry Science 7 (4): , 2008 ISSN Asian Netwrk fr Scientific Infrmatin, 2008 The Effect f Suplemental Ascrbic Acid n the Develpment f Newcastle Disease in Japanese Quail (Cturnix cturnix Japnica) Expsed t High Ambient Temperature B.A. Usman, A.U. Mani, A.D. El - Yuguda and S.S. Diarra 1 Department f Veterinary Medicine, University f Maiduguri, P.M.B. 1069, Brn State, Nigeria 2 Department f Animal Health and Prductin, Mhamet Lawan Cllege f Agriculture, P.M.B. 1427, Maiduguri, Nigeria Department f Veterinary Micrbilgy and Parasitlgy, University f Maiduguri, Maiduguri, Nigeria 4 Department f Animal Health and Prductin, Ybe Cllege f Agriculture Gujba, P.M.B. 1104, Damaturu, Nigeria 3 Abstract: In rder t study the develpment f Newcastle Disease (ND) in Japanese quail subjected t high ambient temperature with r withut supplementatin with ascrbic acid (AA), eighty 3 week-ld Japanese quails were randmly alltted t fur equal grups. Tw f the grups were kept at rm temperature (26±0.5 C) with ne supplemented with AA (R A ) and the ther receiving n AA (R O). The ther tw grups were kept at a cntinuus temperature f 41.8±0.6 C with ne f them supplemented with AA (H A) and ther receiving n AA (H O ). The grup R O served as the cntrl. Ascrbic acid was supplemented at the rate f 1000mg/kg feed. Twelve birds in each f fur grups were challenged culnsally with a pigen islate f velgenic ND virus and the rest left as in cntact. Mrbidity rate was 100% in all challenged and in cntact birds except in the incntact f the grup supplemented with AA which recrded 50%. One hundred percent mrtality was recrded in heat stressed grups irrespective f AA supplementatin while 0% and 50% mrtality was recrded in thse kept at rm temperature with r withut AA supplementatin respectively. Nne f the unchallenged in cntact birds died in any f the grups. Lesins f ND in the experimentally infected quails were characterized by pathlgical lesins. The results f haemagglutinatin inhibitin test perfrmed n bld samples f all birds n days 0, 7, 14 and 21 pst inculatins indicated n effect f heat treatment r AA supplementatin n the humral immune respnse f the Japanese quail. It was cncluded that prlnged high ambient temperature has detrimental effects n the survival f Japanese quail with AA supplementatin having sme amelirating effects. High ambient temperature may nt adversely affect the humral immune respnse f Japanese quail t Newcastle disease and AA may nt be an imprtant factr in the resistance f quails expsed t prlnged high ambient temperature t ND virus infectin. Key wrds: Pathlgy, quail, ascrbic acid, high ambient temperature, Newcastle disease, immune respnse Intrductin The Japanese quail (Cturnix cturnix japnica) have tremendus ptential fr village and backyard prductin as well as an imprtant labratry animal (Lmbin, 2007). This is because f their small size, the ease with which they can be kept, requiring nly simple cages and equipments and little space (Ruskin, 1991), high egg prductin reaching 300 per hen per year (Phillips, 1948; Banks, 1979) and a shrt reprductive life cycle (nly eight weeks are required t hatch eggs and raise the chicks t sexual maturity (Phillips, 1948). They are als rbust, disease resistant and hardy and can easily adapt t many different envirnments. There is paucity f data n the pathlgy f Newcastle disease in quail especially in ht envirnment, even thugh several reprts f utbreaks f Newcastle disease in flcks f Japanese quail have been made (Kaleta and Baldauf, 1988; Chandrasekaran and Aziz, 1989; Islam et al., 1994). High ambient temperature has als been shwn t have an adverse effect n the develpment f immunity in birds (Siegal, 1971; Thaxtn, 1978; Grss and Siegal, 1993) as well as n resistance t diseases, grwth rate and reprductin (Ladewig, 1987). The beneficial effect f ascrbic acid (AA) supplementatin n the perfrmance f birds challenged with stressrs has been reprted (Lin et al., 2006) In this study the effects f ambient temperature and ascrbic acid (AA) supplementatin n the develpment f ND in quails was investigated. Materials and Methds Eighty three-week-ld Japanese quail were randmly divided int fur grups/treatments cntaining 20 birds 328

2 Usman et al.: Effect f Suplemental Ascrbic Acid n the Develpment f Newcastle Disease each. Tw f the grups were expsed t high ambient temperature (41.8±0.6 C), with ne f them supplemented with ascrbic acid (AA) and the ther receiving n AA (H). The ther tw grups were kept at rm temperature (26± 0.5 C) again with ne f them supplemented with AA (RA) and the ther receiving n AA (RO). The grup maintained at rm temperature with n AA (RO) served as reference /cntrl grup. Ascrbic acid (AA) was supplemented at the rate f 1000mg/kg feed. Twelve birds in each grup were challenged culnasally with 0.05ml f pigen strain f velgenic Newcastle disease virus and the remaining 8 birds were left unchallenged as in cntact. Bld was cllected frm the wing vein f all birds fr serlgical examinatin. Birds were bserved fr clinical signs and pstmrtem examinatin was perfrmed n all dead birds The tissues were taken frm lungs, intestine, prventriculus and brain and fixed in 10% frmalin, cut 5mm thick in paraffin wax and stained with H and E (Drry and Wallingtn, 1967) and bserved under light micrscpe at x 80. Results Clinical signs: Clinical signs were first bserved in grup H five days pst-infectin. The majr clinical signs in all affected birds challenged r unchallenged in cntact were dullness, ruffled feathers and respiratry signs in the frm f gasping (plate I), dyspnea and nasal discharge. All the challenged birds in the fur grups except R A challenged exhibited greenish diarrhea and nervus signs in the frm f trticlis, paralysis f legs and wings (plate II). Grup R A challenged exhibited nly respiratry signs. Mrbidity and mrtality: The mrbidity and mrtality f the fur grups are shwn in Table 1. One hundred percent (100%) mrbidity was bserved in all the challenged grups irrespective f their envirnment (rm r high ambient temperature) AA supplementatin and in cntacts birds with n supplemental AA irrespective f envirnmental temperature. Fifty percent (50%) mrbidity was recrded in incntact grups supplemented with AA irrespective f their envirnmental temperature (H A and R A). The vents f the challenged quail that died were siled with greenish faeces. The trachea f birds in the H A grup were heamrrhagic, thse f R were mucid while thse f H shwed n lesin. Three f the dead birds in H grup had slightly inflamed and A heamrrhagic Peyers patches. The air sacs were cludy, the lungs edematus, the prventriculus heamrrhagic and the intestines cngested in all the dead birds (Plate III). Table 1: Mrbidity and mrtality f Japanese quail in the different treatment grups after experimental infectin with a pigen islate f Newcastle disease virus Treatment Parameter (%) RO RA HO HA Mrbidity Challenge Unchallenged Mrtality Challenge Unchallenged RO = rm temperature with n ascrbic acid, RA = rm temperature with ascrbic acid, HO = high temperature with n ascrbic acid, HA = high temperature with ascrbic acid. Table 2: Gemetric mean titre (GMT) value f HI antibdy in Japanese quail in the different treatment grups befre and after experimental infectin with a pigen islate f Newcastle disease virus Days pst infectin Treatments RO Challenged Unchallenged 5.6 -ve RA Challenged Unchallenged 5.6 -ve -ve 80 HO Challenged Unchallenged HA Challenged NT Unchallenged RO = rm temperature with n ascrbic acid, RA = rm temperature with ascrbic acid, HO = high temperature with n ascrbic acid, HA = high temperature with ascrbic acid NT: Nt tested (birds in the grup died befre day 21 pst challenge). Histpathlgical lesins: Lesins were bserved in the brain, lungs, prventriculus and intestine f the dead and dying quail in all the grups except grup RAwhich experienced n mrtality. Brain: Nn suppurative encephalitis characterized by hyperaemia f bld vessels with lymphcytic infiltratin int the Virchw-Rbin s space (Plate IV) was bserved in birds f the three grups that recrded mrtality. Lungs: These exhibited cngestin, haemrrhage, edema and lymphcytic infiltratin int the interstitium (Plate V). Prventriculus: Lesins bserved in this rgan included necrsis f mucsal lining and infiltratin f lymphcytes int lamina prpria (Plate VI). Intestine: Necrsis f mucsal epithelium with lymphcytic infiltratin int the lamina prpria and submucsa were bserved in the intestine.(plate VII) Gemetric mean titre (GMT) value: Table 2 shws The GMT values f Japanese quails challenged culnasally with the pigen virulent islate f ND virus and thse in cntacts. Quails in all the grups recrded psitive ser- 329

vessels. ( H and E), x80. Plate 2: Japanese quail infected with ND virus shwing trticlis and paralysis f legs.

Plate 3: Prventriculus f Japanese quail infected with ND Virus shwing fcal areas (arrw) f heamrrhagic. cnversin just befre challenge.

The highest titre (40) was recrded in the challenged birds f grup H.

3 Usman et al.: Effect f Suplemental Ascrbic Acid n the Develpment f Newcastle Disease Plate 1: Japanese quail infected with ND virus shwing dullness, ruffle feathers and gasping fr air. Plate 4: Nnsuppurative encephalitis in the brain f Japanese quail infected with ND virus shwing lymphcytic perivascular cuffing (arrw) and hyperaemia f bld vessels. ( H and E), x80. Plate 2: Japanese quail infected with ND virus shwing trticlis and paralysis f legs. Plate 5: Lungs f Japanese quail infected with ND virus shwing pneumnitis with cngestin (arrw) and edema. (H and E) x 80. Plate 3: Prventriculus f Japanese quail infected with ND Virus shwing fcal areas (arrw) f heamrrhagic. cnversin just befre challenge. Psitive titre was detected at 7 day pst infectin (PI) in all grups except in cntacts at rm temperature (RA and R ). The highest titre (40) was recrded in the challenged birds f grup H. At 14 days PI ser-cnversin was als recrded in all the grups except in the unchallenged in cntact f R A grup, the challenged half f which recrded the highest (28.3) titre. By the 21st day after challenge all challenged birds in grup HA died while all ther grups, bth challenged and unchallenged in cntacts, recrded their highest ser-cnversin f the experiment. Discussin The incubatin perid f 5 days, clinical signs, grss and histpathlgical lesins bserved in quail infected with the velgenic pigen strain f ND virus and thse in cntact are similar t thse reprted in quail and ther avian species elsewhere (Hills et al., 1953; Kumanan et al., 1990; Crss, 1991; Emmersn, 1994; Lam, 1996; Sa idu et al., 2006; Oladele, et al., 2008). 330

Usman et al.: Effect f Suplemental Ascrbic Acid n the Develpment f Newcastle Disease Plate 6: Nnsuppurative prventriculitis in Japanese quail infected with ND virus. Nte.

Plate 7: Intestine f Japanese quail infected with ND virus shwing necrsis f villi (arrw) and leuccytic infiltratin. ( H & E) x80.

4 Usman et al.: Effect f Suplemental Ascrbic Acid n the Develpment f Newcastle Disease Plate 6: Nnsuppurative prventriculitis in Japanese quail infected with ND virus. Nte. Mucsal epithelial necrsis(arrw) with mnnuclear cell infiltratin in the lamina prpria and submucsa (H & E) x80. Plate 7: Intestine f Japanese quail infected with ND virus shwing necrsis f villi (arrw) and leuccytic infiltratin. ( H & E) x80. The results f this study clearly demnstrated the susceptibility f quail t infectin with ND virus challenged culnasally and direct cntact with infected birds. The higher (100%) mrtality recrded in all the challenged grups suggest that the susceptibility f quail t the ND virus appeared t be nt influenced by heat stress r AA supplementatin. Ascrbic acid (AA) supplementatin hwever, appeared t have beneficial effect n the mrbidity f the disease in incntact birds irrespective f heat stress. The lwer mrtality rate recrded in nn heat stressed cmpared with heat stressed quails indicates the detrimental effects f high envirnmental temperature n the utcme f ND virus infectin. This bservatin supprts the view f Thaxtn, (1978) that the envirnment influences birds ability t withstand infectius diseases. The absence f any difference in mrtality rates between AA supplemented and unsupplemented grups f the heat stressed quails suggests that AA supplementatin has n beneficial effects in reducing mrtality due t ND virus infectin in the heat stressed birds. The results f gemetric mean titre (GMT) determinatin in this study suggest that heat stress did nt interfere with humral immune respnse f Japanese quails challenged with ND virus since all the challenged and unchallenged in cntact birds develped similar antibdy titres. The levels f humral immune respnse hwever were nt sufficient t cnfer prtectin against virulent challenge by virulent ND virus since 100% mrbidity was recrded in all the challenged grups. The lack f significant disparity in immune respnse between heat-stressed and nn-heat-stressed quail culd be assciated with the inherent ability f the Japanese quail t regulate ambient temperatures. Similar studies in chickens have als shwn that circulating antibdy cncentratins were nt affected by high temperature when chickens were expsed t chrnic heat treatment (Thaxtn et al., 1968; Thaxtn, 1978). Thaxtn (1978) hwever, demnstrated that established antibdy levels culd be reduced within a relatively shrt perid f time fllwing an acute heat treatment. This effect was defined by Thaxtn and Siegal (1973) as high envirnment temperature - mediated immundepressin (HTD). The ability f quail t survive the infectin with this virulent ND virus especially in in-cntacts grups despite similar humral respnse with thse that died suggests the pssibility that there are ther mechanisms fr cnferring immunity in infected birds. It was cncluded that ascrbic acid is nt an imprtant factr in the resistance f quail expsed t high ambient temperature t Newcastle disease virus infectin. Hwever, in quail that are nt expsed t elevated temperature, the presence f AA in their diet enhances their resistance t infectin with Newcastle disease virus. References Banks, S., The Cmplete Handbk f Pultry Keeping. Wadlck Ltd., pp: 109. Chandrasekaran, S. and H.A. Aziz, Outbreak f Newcastle disease in Japanese quail. J. Vet. Malay, 1: Crss, G.M., Newcastle disease. Vet. Clin. N. Am. Small. Anim. Pract., 2: Emmersn, P.T., Newcastle disease virus. In: Encyclpaedia f virlgy. Rberts, G.W. and G. Allan (Eds.). Acad. Press Harcurt and Brace Publ., 1: Grss, W.B. and H.S. Siegal, General aspects f stress and welfare. In: Livestck Handling and Transprt. Grandin, T. (Ed.) Willingfrd, U.K. CAB Int., pp:

5 Usman et al.: Effect f Suplemental Ascrbic Acid n the Develpment f Newcastle Disease Hills, D.H., D.S. Davis and J.K.H. Wilde, Lmbin, L.H., Natinal Veterinary Research Newcastle disease in Nigeria. Br. Vet. J., 109: 381- Institute, Vm. Its activities and pprtunities t 385. veterinary prfessinals Paper presented at the Islam, H.A., J. It, H. Tanakuwa, A. Takada, C. Itrakura, 44th Annual cngress f the Nigerian veterinary and H. Kida, Acquisitin f pathgenicity f Medical Assciatin, Effurun, Delta state, Nigeria, Newcastle disease virus islated frm Japanese Octber 22nd -26th. quail by intracerebral passage in chickens. Jap. J. Phillips, L.S., Pultry feed and nutritin. The AVI Vet. Res., 42: Publ. C. Inc. Cnnecticutt, pp: Kaleta, E.F. and C. Baldauf, Newcastle disease in Ruskin, F.R., Quail. In: Micrlivestck: Little Knwn free-living and pet birds. In: Newcastle disease. Small Animals with Prmising Ecnmic Future. Alexander D.J. (Ed). Nrwell, M.A., Kluwer Acad. BOSTID, Natinal Research Cuncil, Natinal Publ., pp: Academic Press, Washingtn, DC., pp: Kumanan, K., D. Thyagarajan, K.S. Palaniswamy, Sa idu, L., P.A. Abdu, L.B. Tekdek, J.U. Umh, M. Usman Sundararasu and K.A. Venkatesan, Ranikhet and S.B. Oladele, Newcastle disease in disease (Newcastle disease) vaccinatin Nigeria. Nig. Vet. J., 27: prgrammes fr Japanese quail. Ind. J. Anim. Sci., Siegal, H.S., Adrenals, Stress and envirnment. 60: Wld. Pult. Sci. J., 27: Ladewig, J., Endcrine aspect f stress: Thaxtn, P., Influence f temperature n the Evaluatin f stress reactins in farm animals. In: immune respnse f birds. A Rev. Pult. Sci., 57: Bilgy f Stress in Farm Animals. An Integrated Apprach. P.R. Weipkema and P.W.M. Van Adrichen Thaxtn, P. and H.S. Siegal, Mdificatin f high (Eds.) Martinus Nijhaff Drdrecht, pp: temperature and ACTH mediated Lam, K.M., NDV-induced apptsis in the immunsupressin by metryrapne. Pult. Sci., 52: peripheral bld mnnuclear cells f chickens. J Cmp. Pathl., 114: Thaxtn, P., C.R. Sadler and B. Glick, Immune Lin, H., H.C. Jai, J. Buyse and E. Decuypere, respnse f chicken fllwing heat expsure r Strategies fr preventing heat stress in pultry. injectin f ACTH. Pult. Sci., 47: Wrld s Pult. Sci., J., 62:

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