Archives of Razi Institute
Volume:56 Issue: 1, Summer 2003

The thymidine kinase (TK) gene of four Iranian herpes simple.\; virus-I isolates was ampli lied and the electrophoretic pallerns or the gene were compared after digestion with six restriction endonucleases including Apal, Aval. Bg/I, Drall. EcoRV and Pst!. DN8A of each virus was 1irst extracted scparately l1y Iysis butTer and phenol/chloroform method. Their TK genes WC1'<: ampli lied lIsing a pair or special primers. which were designed by Gene 8RlInner software. The polymerase chain reaction product of eaeh TK gene was digested with the restriction enzymes separately and electrophoresed in polyacrylamidc gel. The results indicatc that there was no diffcrcnce among the c1ectrophoretic pattcrns or the isolates TK gene.

Tht: clinical signs and postmortem lindings including rt:spiratory system distress and Icsions in upper respiratory tract wert: observed in 17 commercial layer flocks l'rom differcnt parts of Iran. Laryngotracheal tissues l'rom the alTeetcd birds with suspectcd inlectious laryngotrachitis (lLT) wcrc examined l'or deteetion of the causative agt:nt using virus isolation (VI), agar gd imlllunodilTusion (AGIO), histopathology (HP), and polylllcrase chain reaction (peR). 12 (70.5%) of the salllples were formed typical poeks on chrioallantoic membrane of embryonated specilic pathogen free eggs in VI method. By AGIO test Il (64.7%) of the samples \Vere detectcd as II.T virus (1I.Tv) that gave c1ear lines \Vith hyperimmune IL T serum. Of the samples 7 (41.1 %) were illustrated lesions chara\.:teristics of ILT including inliltration of inllammatory œlls and syncytia formation. In 14 (82.3%) or the salllples ILTv was dcteetcd h) l'CR. The high positive perccntages of the l'CR indieate that the technique is applicable in rapid diagnosis of ILT for its accurucy, sensitivity and specilieity.

Experimental oil-emulsion vaccinc \Vas l'onnulated \Vith a ratio of one part of 0.05% ~-propiolactonc inactivatcd antigcn and l'our parts oil adjuvant ISA-70. Four hundn.:d whitc Ily-Linc (\\'-36) chicks were rearcd in t'our groups. Groups 1 and 2 \Vere vaccinated b) live attenuated 1·1120 at day 10 and reared separately from groups 3 and 4. At weck 16 groups 1 and 2 were received experirnental oil-emulsion and commercial oil-emulsion vaccines respectively. At this time. group 3 \Vas n:ceivcd single dose of experimental oil-emulsion vaccine and group 4 \\"s unvaccinated conlrol. At \Vcck 29 ail or groups \\cre challenged by 0.25' I07r': ID,,/bird or intCcted allantoic Iluid whcn the)' were on peak of production. Ali groups were bled fi-cl]uently and the sera were assayed by ELISA and ACJI J) tests. Clinical signs and high percentage drop in egg production in groups 3 and 4 v.ere notcd. Moreover. no virus and/or viral antigen in the trachea or groups 1 and 2 \Vere dctcctcd at days 5 and 10 postchallengc. The results or clinical observation cgg producl ion. virus isolation and delcction in Ihe trachea and levels of antibody suggcsted Ihal layers vaecinated wilh a combinalion of live allcnuated and the cxpcrimcntal inactivalcd vaccines had Ihc highcsi proteclion.

The isolation of viral causalive agenl or fry lrout morlality syndrome in farmed rainbow troul in Iran was discussed. Four hundred twcnty samples were eollected from six provinces for virological, serological and clectron microscopie examinations from October 2001 lill May 2002. No eylopathic elTeet (CPE) \Vas observed in the cell cultures excepl for EPC and BF-2 cclls in whieh signs of CPE appeared one day postinoculalion during lhe second and lhird passages or Iwo brooder ovarian fluids. I.:leclron microscopie examination revcaled the viral partieles measured 67-73 x 128-l82nm in lhe pelleted materials from the CPE positive in 81'-2 and EPC cell lines. Indirecl immunoflurccence antibody test \Vere positive lor sorne ot" smears obtained From kidney tissues of lhe larvae, the samples of ovarian lluids lhal showed CPE on EPC and BF-2 ccII lines as weil as CPE positive œil cultures. Using U.lS,\ eighl of ail sera samples lcslcd \Vere posilive \Vilh oPlieal dcnsil) rangcd (U 16-2.268 compared 10 0.06 f()r negali\'c conlrols.

By emergence of very virulent infectiolls bursal disease virllses (vvIBOVs), ail classical measures were called into question. In this study an inactivated IBO vaccine with bursal and embryo origins using a recent local vvlBOV isolate, 1R499, was prepared. The virus cultivated in both SPF emberyonated eggs and chicks to evaluate the efTects of host system on immunogenicity of the vaccine. The results showed that inoculation of broiler breeder chickens with the inactivated IBO vaccine at 20 weeks of age produced significantly greater levels of antibody in comparison with IBO live vaccine and induced antibody titers with a lower variation than the other. The mean antibody titer raised by bursal origin vaccine was significantly greater than other inactivated vaccine and it persisted throughout this study. Our result show that inactivated vaccine with bursal origin induces high level of antibodies, which is essential for protecting of the birds against the vvIBOV strain. Also il shows that incillding of very virulent strain in inactivated vaccine provides bcttcr protection against lield vvlBOV strains.

Various cell wall rractions of Pasleurella mu/toeida vaccine strain serotype A: 1 including sonicated antigen, heat stable (HS), Iipopolysaccharide-protein complex (LPS-P). olltermembrane protein (OMP), capsular (CAP) and potassium thiocyanate (KSCN~xtracted proteins were extracted. Their immllnogenecity was studied and compared with inactivated whole cell vaccine in vitro and in vivo. The protein estimation and the polypeptide pattern of these antigens were also compared by Lowery and SOS-PAGE methods. In general the immunological responses to these fractions were varied. The chickens immunized with KSCN-extract antigen and OMP showed the highest protection when they were challenged with homologous strain, while ifS and LPS antigens showed the lowest protection, These findings are in correspondence with the end-point titer of antibodies using ELISA. Cross-reaction was also observed by agar gel difussion test between the fractions and raised antisera in chickens especially with KSCN-extract. OMP and CAP antigens, The results suggcst that both OMP and KSCNextract protein as more effective immllnogens cOlild be good candidates for devclopmcnt or a subunit vaccine substitut\: for current who le œil formalized vaccine against fowl cholera infection.

IFN-y and IL-4 levels of peripheral blood mononuclear ccli (PBMC) cultures and sera of Ihe cyslic echinococcosis palienls wilh primary and relapsed infeclions were compared. In Ihis regard. PB MC of Ihe patienls and controls were eullured in RPMI and stimulaled wilh different hydatid Iluid antigens of (SHf. pI-l5PPT, B-Ag, FI and F2). IFN-y and IL-4 of the culture supernatanls as weil as the sera of echinoeoecosis palienls wen: evaluated by ELISA. The results indicale thal the level of IL-4 produced by PBMC of the patients was higher Ihan thal of heallh)' group and the diffen::ncc was signifieant. The levels of IL-4 in PBMC eullure and sera of Ihe palienls with relapsed hydatidosis were significantly higher th an Ihose wilh primary infection means the immune response paltern of the patients. who have nol receiwd any anliparasitic drug aller surgery. ma)' shift loward Th2 activalion and suseeplibilily 10 Ihe infeclion. Contrar)'. the levels of IFN-y produccd bl' PBMC and in sera of Ihe palients Ircaled surgicall) once do not di/Ter significantly with Ihose tn:ated two limes or more.

Changes in diamctcrs of the prc:ovulatory dominant follicle (l'or 10 days bel'on: 0\ ulation) and corpus luteum (during the estrous cycle) parallcling \\·ith plasma estradiol-17~ and progesterone profiles \Vere monitored in sevcn healthy Caspian marcs. agcd 8.8± 1.53 years and weighed 198.6±0.9kg. The plasma cnnccntration of estradiol-17~ \Vas progressively (1'<0.05) increased fi'om day (, to day 2. when the peak value \Vas attained. [1 then sharply decreased to the basal concentrations at the time of ovulation. The tirsl signiticanl (1'<0.05) risc in plasma progesterone concentration was delected 24h alter ovulation. It \\a5 steadil)' incrcased until day 4 arter ovulation. The concentrations IVere rcmained elevated unlil day 12. From day 10 onward the graduai decn:<1se in plasma progesterone concentrations was noticed with the !irst signliicalll drop IKt\\een days 12 and 13. There lVas a signilicant (1'<0.05) correlation bct\\'een plasma progesterone and estradiol-17~ concentrations with cyclic changes in corpus luteum and preovulatory [ollicle. respectivel} The results suggest that in Caspian mares as compared \0 other breeds. the progestérone Icvels began to decrease earlier. due to earlier occurrence of' luteolysis. and regarding t

The osteogenicity and crrect on early bone repair of bone marrow grafls were invcstigated. The purposc of this expcrimcntal study was to determine whl!thcr bonc marrow gralied pcrcutaneously Icads to increased bone production or has any I!ITcct on the early healing of fractures. Thirty adolescent. male white rabbits were used. Ali of them had bilateral radial osteotomies. Rabbits were divided to equal five groups. The cross-sectional are a of cali us. breaking load. tensile strength and callus volume parameters at the rra Win: site wcre tested. At two weeks postgrafting callus volume were signilicantly higher (0001<1'<0005) in grafted radii than in the contralatcral saline control. Gy four wccks ail parameters were signilicantly greater in the bone marrow gratled radii th an in the control. Seriai radiographs ancl histology conlirm this advanced fracture healing in the gral1cd bones. There were no difkrcnces between the external callus of treated and control radii but the internai callus bctween the end of the cortical bone did show a clilTcrence bdwCl:n the two sides. Pcrcutaneous bone marrow grafLing is a simple scmiinvasile techniquc that ma)' have potential clinical application.

l'hl: biological activity of the rl:combinant growth hormone (GIll s)'nthesiled in The National Researeh Center l'or Genetic Engineering and Biotcchnology (NRCGEB) \Vas evaluated in vivo. In this regard a hypophysectomized rat modcl was required. Using a para pharyngeal approach. the sphenoid bone of rats \Vas drilled and the hypophysis was out succcssfully by aspiration. Following a 7-day rccovery period. rats \Vere in je ct cd suhculaneously cithcr \Vith the recombinanl human GH or the Novo. as standard. The resulls show lhal both hormones havc an equal hilliogicai aClivit) and resulled in a 100% growlh incrcmcnl equal to that in untrealed inlact rats indicaling Ihat Ihe recombinant GII can be introduccd as a good replacemcnt of endogenous GIf