Archives of Razi Institute
Year:2003 | Volume:- | Issue:56|Papers Count:10

The thymidine kinase (TK) gene of four Iranian herpes simplex virus-I isolate was amplified and the electrophoretic patterns of the gene were compared after digestion with six restriction endonucleases including ApaI, Bg/I, DraII, EcoRV and PstI. DN8A of virus was first extracted separately by lysis buffer and phenol/chloroform method. Their TK genes were amplified using a pair of special primers, which were designed by Gene 8Runner software. The polymerase chain reaction product of each TK gene was digested with the restriction enzymes separately and electrophoresed in polyacrylamide gel. The results indicate that there was no difference among the electrophoretic patterns of the isolates TK gene.

The clinical signs and postmortem finding including respiratory system distress and lesions in upper respiratory tract were observed in 17 commercial layer flicks from different parts of Iran. Laryngotracheal tissues from the affected birds with suspected infectious laryngotrachitis (ILT) were examined for detection of the causative agent using virus isolation (VI), agar gel immunodiffusion (AGID), histopathology (HP), and polymerase chain reaction (PCR). 12 (70.5%) of the samples were formed typical pocks on chorioallantoic membrane of embryonated specific pathogen free eggs in VI method. BY AGID test 11 (64.7%) of the samples were detected as ILT virus (ILTV) that gave clear lines with hyperimmune ILT serum. of the samples 7 (41.1%) were illustrates lesions characteristics of ILT including infiltration of inflammatory cells and syncytia formation. in 14 (823%) of the PCR indicate that the technique is applicable in rapid diagnosis of ILT for its accuracy. sensitivity and specificity.

Experimental oil- emulsion vaccine was formulated with a ration of one part of 0.05% β- propiolactone inactivated antigen and four parts oil adjuvant ISA -70. Four hundred white. HY - Line (W-36) chicks were reared in four groups. Group 1 and 2 were vaccinated by live attenuated H 120 at day 10 and reared experimental oil -emulsion and commercial oil - emulsion vaccines respectively. At this time group 3 was received single dose of experimental oil -emulsion vaccine and group 4 was unvaccinated control. At week 29 all of groups were challenged by 0.25.107 EID50 bird of infected allantoic fluid when they were on peak of production. All groups were bled frequently and the sera were assayed by ELISA and AGLD tests. Clinical signs and high percentage drop in egg production in groups 3 and 4 were noted moreover. On virus and or viral antigen in the trachea of groups 1 and 2 were detected at days 5 and 10 postchallenge. The results of clinical observation egg production virus isolation and detection in the trachea and levels of antibody suggested that layers vaccinated with a combination of live attenuated and the experimental inactivated vaccines had the highest protection.

The isolation of viral causative agent of fry trout mortality syndrome in farmed rainbow trout in Iran was discussed. Four hundred twenty samples were collected from six provinces for virological. Serological and electron microscopic examinations from October 2001 till may 2002. No cytopathic effect (CPE) was observed in the cell cultures except for EPC and BF - 2 cells in which signs of CPE appeared one day postinoculation during the second and third passages of two brooder ovarian fluids Electron microscopic examination revealed the viral particles measured 67-73 ×128-182 nm in the CPE positive in BF - 2 and EPC cell lines. Indirect immunoflurecence antibody test were positive for some of smears obtained from kidney tissues of the larvae. The samples of ovarian fluids that showed CPE on EPC and BF-2 cell lines as well as CPE positive cell cultures Using ELISA eight of all sera samples tested were positive with optical density ranged 0.316 -2 268 compared to 0.06 for negative controls.

By emergence of very virulent infectious bursal disease viruses (vvIBDVs), all classical measures were called into question. In this study an inactivated IBD vaccine with bursal and embryo origins using a recent local vvIBDVs isolate IR 499 was prepared. The virus cultivated in both SPF emberyonated eggs and chicks to evaluate the effects of host system on immunogenicity of the vaccine. The results showed that inoculation of broiler breeder chickens with the inactivated IBD vaccine at 20 weeks of age produced sigificatly greater levels of antibody in comparison with IBD 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 shoe that inactivated vaccine with bursal origin induces high level of antibodies. Which is essential for protecting of the birds the vv IBDV strain? Also it shows that including of very virulent strain in mactivated vaccine provides better protection against fleld vv IBDV strains.

Various cell wall fractions of Pasteurella multocida vaccine strain serotype A.1 including sonicated antigen heat stable (HS) lipopolysaccharide - protein complex (LPS - P ) outermenbrane protein (OMP) capsular (CAP) and potassium thiocyanate (KSCN) - extracted proteins were extracted their immunogenecity 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 SDS-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 HS 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 diffusion test between the fractions and raised antisera in chickens especially with KSCN - extract OMP and CAP antigens the results suggest that both OMP and KSCN - extract protein as more effective immunogens could be good candidates for development of a subunit vaccine substitute for current whole cell formalized vaccine against fowl cholera infection.

IFN - γ and IL - 4 levels of peripheral blood mononuclear cell (PBMC) cultures and sera of the cystic echinococcosis patients with primary and relapsed infections were compared in this regard PBMC of the patients and controls were cultured in RPMI and stimulated with different hydatid fluid antigens of (SHF. PH5PPT. B-Ag, F1 and F2) IFN γ and IL - 4 of the culture supernatants as well as the sera of echinococcosis patients were evaluated by ELISA. The results indicate that the level of IL - 4 produced by PBMC of the patients was higher than that of healthy group and the difference was significant. The levels of IL - 4 in PBMC culture and sera of the patients with relapsed hydatidosis were significantly higher than those with primary infection means the immune response pattern of the patients who have not received any ant parasitic drug after surgery, may shift toward th2 activation and susceptibility to the infection. Contrary the levels of IFN γ - produced by PBMC and in sera of the patients treated surgically once do not differ significantly with those treated two times of more.

Changes in diameters of the preovulatory dominant follicle (for 10 days before ovulation) and corpus luteum (during the estrous cycle ) paralleling with plasma estradiol - 17 β and progesterone profiles were monitored in seven healthy Caspian mares aged 8.8± 1.53 Years and weighed 198.6 ± 0.9 kg . The plasma concentration of estradiol - 17 β was progressively (p<0.05) increased from day 6 to day 2. When the peak value was attained, it then sharply decreased to the basal concentrations at the time of ovulation, the first significant (p<0.05) rise in plasma progesterone concentration was detected 24 after ovulation, it was steadily increased until day 4 after ovulation. the concentrations were remained elevated until day 12 from day 10 onward the gradual decrease in plasma progesterone concretions was noticed with the first significant drop between days 12 and 13 there was a significant (P<0.05) correlation between plasma progesterone and estradiol - 17 β concentrations with cyclic changes in corpus luteum and preovulatory follicle respectively the results suggest that in Caspian mares as compared to other breeds. The progesterone levels began to decrease earlier. Due to earlier occurrence of autolysis and regarding to estradiol - 17 β there was an extra minor peak that occurred at about day 8.

The osteogenicity and effect on early bone repair of bone marrow grafts were investigated. The purpose of this experimental study was to determine whether bone marrow grafted percutaneously leads to increased bone production or has any effect on the early healing of fractures. Thirty adolescent, male white rabbits were used. All of them had bilateral radial osteotomies. Rabbits were divided to equal five groups. The cross-sectional area of callus, breaking load, tensile strength and callus volume parameters at the fracture site were tested. At two weeks postgrafting callus volume were significantly higher (0.001<P<0.005) in grafted radii than in the contralateral saline control. By four weeks all parameters were significantly greater in the bone marrow grafted radii than in the control. Serial radiographs and histology confirm this advanced fracture healing in the grafted bones. There were no differences between the external callus of treated and control radii but the internal callus between the end of the cortical bone did show a difference between the two sides. Percutaneous bone marrow grafting is a simple semiinvasive technique that may have potential clinical application.  

The biological activity of the recombinant growth hormone (GH) synthesized in the national research center for Genetic Engineering and Biotechnology (NRCGEB) was evaluated in vivo. In this regard a hypophysectomized rat model was required. Using a Para pharyngeal approach the sphenoid bone of rats was drilled and the hypophysis was out successfully by aspiration following a 7- day recovery period rate were injected subcutaneously either with the recombinant human GH or the Novo as standard . The results shoe that both hormones have an equal biological activity and resulted in a 100% growth increment equal to that in untreated intact rats indicating that the recombinant GH can be introduced as a good replacement of endogenous GH.