فهرست مطالب

Archives of Razi Institute
Volume:73 Issue: 3, Summer 2018

  • تاریخ انتشار: 1397/06/05
  • تعداد عناوین: 9
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  • M. Mehrabi, H. Montazeri, N. Mohamadpour Dounighi , A. Rashti, R. Vakili, Ghartavol Pages 165-176
    Most infectious diseases are caused by pathogenic infiltrations from the mucosal tract. Nowadays, the use of vaccines has been widely investigated for the prevention of different infectious diseases, infertility, immune disorders, malignancies, and allergies. Broad-spectrum adjuvant substances have been studied for immune system stimulation with a greater efficiency against specific antigens. Various adjuvants have been developed such as inorganic, oil-based, and emulsion adjuvants, bacterial products and their derivatives, cytokines, cytosine-guanine dinucleotide (CpG) motifs, and particulate systems. Mucosal vaccine delivery is an alternative route to induce both humoral and cellular immune responses. Applying nanoparticles in vaccine formulations allows not only improved antigen stability and immunogenicity, but also targeted delivery, and consequently, more specific release of the agent of interest. Chitosan nanoparticles have immunological activity and mucoadhesive properties. They have been used as a mucosal vaccine delivery system for many antigens. This review provides an overview of the recent advances in chitosan nanoparticles as a novel mucosal vaccine delivery system.
    Keywords: Adjuvant, Chitosan Nanoparticle, Mucosal Vaccine Delivery
  • S. G. Mirzaei, A. Shoushtari , A. Nouri Pages 177-182
    Avian influenza H9N2 subtype viruses have had a great impact on Iranian industrial poultry production economy since introduction in the country. To approach Rapid and precise identification of this viruses as control measures in poultry industry, a real time probe base assay was developed to directly detect a specific influenza virus of H9N2 subtype -instead of general detection of Influenza A viruses- which has been endemic over two last decades in the country. An Iranian avian influenza virus strain of A/Iran/chicken/772/1998 H9N2 subtype were selected as reference strain for of primers and probe designing. The high agreement value of 99% indicated that the devolved real time assay for detection of H9 subtype viruses could easily replace the conventional method of virus isolation particularly in investigation of viruses like national surveillance plan. The limit of detection was almost one EID50 which was the least real infectious unit could be detected. So it can be said that this sensitive assay provided a powerful tool to not to miss any significant viral biological activity neither in the host body nor in the environment. A high level of correlation coefficient (R2 = 0.998) also indicated a good correlation between Ct values and viral concentrations. , it can be conclude that the real time RT-PCR could be easily replace virus isolation in detection of H9N2 influenza viruses especially in large monitoring program. The ability in quantifying of the virus concentration extends usage of test in more accurate studies.
    Keywords: Real-Time RT- PCR, Avian influenza, Comparison, H9N2, Culture
  • M. Vasfi Marandi, M. Malekan , M. M. Ranjbar, N. Dadashpour Davachi , S. Alamian Pages 183-198
    Infectious bronchitis (IB) is an acute, highly contagious, and economically important viral disease of chickens. The S1 subunit from Spike (S) protein plays the major role in protective immunity and is involved in the host-virus interactions, as well as infectious bronchitis virus (IBV) serotyping. Aim of the present study was multi-aspect analysis of the molecular and immunological features of 5' part belonging to the S1 glycoprotein sequence of Iranian 793/B IBV strain isolates. This might ideally help in characterization, prevention, and vaccine development. The tissue samples were prepared, followed by virus isolation, reverse transcription polymerase chain reaction and restriction fragment length polymorphism analysis. In addition, sequencing and registration of the sequences in the National Center for Biotechnology Information were performed. Moreover, 12 sequences were retrieved from Fars province, Iran. The next steps included evaluation of conservation/variability along the sequences, phylogenetic analysis, estimation of the average evolutionary divergence over all the sequence pairs, predicting the phosphorylation/N-glycosylation/palmitoylation sites, and the final analysis of antigenicity. The findings of alignment, entropy plot, and pairwise similarity analysis revealed 17 hypervariable regions. The isolates belonging to Tehran were clustered in phylogenetic tree, and the most similar isolates to them were ADW11182 and ADW11183. Location of some of the N-glycosylation/phosphorylation/palmitoylation points indicated that these sites were conserved among the isolates. Furthermore, the frequency of epitopes and their scores reflect the high immunogenicity of S1 protein in 793/B serotype. Analysis of the primary and secondary structures demonstrated that their parameters had variable values and were different regarding the number and location of α-helix, β-strand, and coils. According to our findings, the Iranian isolates of 793/B serotype change their molecular characteristics during time and in different geographical regions. These alterations might account for failure in prevention programs and differences in virulence and pathogenicity
    Keywords: 793, B Serotype, bioinformatics, Infectious Bronchitis Virus, Molecular features, S1 glycoprotein
  • M. Naghavi, M. H. Sekhavati , M. Tahmoorespur, M. R. Nassiri Pages 199-206
    Brucellosis is a zoonotic disease in human and animals. Brucella melitensis is one of the most pathogenic species of Brucella in goat and sheep. Omp31 is an outer membrane protein of Brucella that acts as an immunogenic protein. Cytokines are glycoproteins with low molecular weight that play the role of an immune adjuvant and regulate immune responses. Interleukin-2 is one of the most important cytokines, which are secreted by the white blood cells and involved in T cell immune responses. In the present study, a chimeric Omp31-Interleukin2 recombinant protein was generated by means of genetic engineering techniques. This chimeric coding sequence was amplified by using specific primers and using Splicing Overlap Extension (SOE) PCR technique. The fusion of the two mentioned proteins was accomplished using a rigid linker. The generated chimeric IL2-Omp31 fragment was TA cloned, and then subcloned into pEt22b vector as an expression vector. The chimeric protein was successfully expressed in E. coli BL21 (DE3) and confirmed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and also Western-blotting analysis. Finally, in order to assess the antigenic features of the recombinant chimeric IL2-Opm31 protein, its secondary structure and antigenicity were predicted in silico.
    Keywords: Brucella melitensis, Interleukin, Omp31, Cytokines
  • J. Soodmand, T. Zeinali , G. Kalidari, G. Hashemitabar, J. Razmyar Pages 207-213
    Enterococci are Gram-positive facultative anaerobic bacteria commonly found in the gastrointestinal tract of the mammals and birds. These cocci are isolated from urinary tract infections, bacteremia, endocarditis, and burn wounds in humans. The evolution of antibiotic-resistant enterococci raised a problem due to the possibility of the transmission of these organisms between poultry and human. Regarding this, the present study was conducted to evaluate the prevalence of Enterococcus species among companion birds and poultry in the Northeastern of Iran and determine the antibiotic susceptibility profile of enterococci. To this end, oral and cloacal swabs were collected from 150 caged birds. Antibiotic susceptibility profile was determined using the standard disk diffusion method. The results revealed that out of 150 samples, 56 (37.33%) cases contained enterococci. Most of the specimens (25.33%) were Enterococcus faecalis isolated from 6.66% of the samples. Additionally, 2.66% and 1.33% of the samples were contaminated with Enterococcus mundtii and Enterococcus gallinarum, respectively. Furthermore, Enterococcus malodoratus and Enterococcus raffinosus were isolated from 0.66% of the samples. The results revealed that all of the isolates of E. faecalis and E. faecium were resistant to more than five antimicrobial agents. Most of E. faecalis and E. faecium isolates showed resistance to Cefazolin, Tiamulin, Flumequine, and Cephalexin. Accordingly, the majority of the isolates had multidrug resistance to the tested antibiotics. In conclusion, the presence of multidrug-resistant enterococci in the birds living close to humans requires thorough observations due to the transmission of these organisms to humans.
    Keywords: Enterococci, Bird, Antibiotic resistance
  • B. Mosallanejad , S. Bahrami, H. Hamidinejat, S. Ghanavati Pages 215-221
    Dogs are important in the epidemiology of Neospora caninum because they act as definitive hosts, shedding oocysts in the environment. The aim of the present survey was to evaluate the serological prevalence of Neospora caninum infection in urban and rural dogs in Ahvaz district, southwest of Iran. In this study, blood samples were taken from 100 rural dogs and 50 urban dogs. The dogs were categorized into two age groups (i.e., ≤ 3 and > 3 years). Neospora agglutination test (NAT) was performed for the detection of infection. Among 150 samples, 30 (20%) showed infection in 1:50 to 1:800 dilutions by NAT (confidence interval 95%: 13.60-26.40). The antibody titers were as follows: 1:50 (n=1), 1:100 (n=14), 1:200 (n=3), 1:400 (n=10) and 1:800 (n=2). The highest serum dilution was 1:100 in 46.67% of the infected dogs and the lowest serum dilution was 1:50 in 3.33% of them. The obtained results showed a significant difference in seroprevalence between urban (10%) and rural (25%) dogs (P=0.03). Although the seroprevalence was higher in dogs above three years of age (23.33%) than below three years (17.78%), there was not a significant difference among different age groups in this regard (P>0.05). The possibility of infection in dogs above the age of three years was 1.3 more than those below three years of age (confidence interval 95%: 0.58-2.9). It can be concluded that a relatively considerable percentage of dogs in Ahvaz district are infected with N. caninum. These infected dogs can play an important role in the transmission of neosporosis to other animals.
    Keywords: Neospora caninum, Neospora agglutination test (NAT), Seroprevalence, Ahvaz, Dog
  • H. Hajizadeh, G. Abedi , A. Asghari, S. Hesaraki Pages 223-227
    The present study was conducted with the aim of comparing the effects of premedication with ketoprofen and midazolam in birds. A total of 24 male pigeons with an approximate weight of 300 g were divided into four equal groups. The control group (Group I) was injected with ketamine alone. Groups II-IV were injected with ketoprofen alone, ketoprofen+ketamine, and midazolam+ketamine, respectively. The biochemical changes in the four groups were evaluated after intramuscular drug injections at different anesthetic levels. A record of biochemical changes was maintained for each group. Blood samples were taken before and after the administration of the medications in order to measure the levels of serum alkaline phosphatase (ALP), oxaloacetate transaminase (OT), prothrombin time (PT), glucose (GLU), lactate dehydrogenase (LDH), albumin (Alb), total protein (TP), and gamma-glutamyl transferase (GGTF). The results showed significant differences in the mean levels of ALP, OT, PT, GLU, LDH, Alb, and TP after anesthesia, compared to that before anesthesia. Therefore, ketoprofen+ketamine can be used for the induction of anesthesia in birds.The present study was conducted with the aim of comparing the effects of premedication with ketoprofen and midazolam in birds. A total of 24 male pigeons with an approximate weight of 300 g were divided into four equal groups. The control group (Group I) was injected with ketamine alone. Groups II-IV were injected with ketoprofen alone, ketoprofen+ketamine, and midazolam+ketamine, respectively. The biochemical changes in the four groups were evaluated after intramuscular drug injections at different anesthetic levels. A record of biochemical changes was maintained for each group. Blood samples were taken before and after the administration of the medications in order to measure the levels of serum alkaline phosphatase (ALP), oxaloacetate transaminase (OT), prothrombin time (PT), glucose (GLU), lactate dehydrogenase (LDH), albumin (Alb), total protein (TP), and gamma-glutamyl transferase (GGTF). The results showed significant differences in the mean levels of ALP, OT, PT, GLU, LDH, Alb, and TP after anesthesia, compared to that before anesthesia. Therefore, ketoprofen+ketamine can be used for the induction of anesthesia in birds.
    Keywords: Serum Analysis, Ketamine, Ketoprofen, Midazolam, Pigeons
  • M. Gholizadeh , M.R. Nassiry, M. R. Saberi, A. A. Haddad, Mashadrizeh Pages 229-237
    Phytase increases the bioavailability of phytate phosphorus in seed-based animal feeds and reduces the phosphorus pollution of animal waste. Since most animal feeds for pellets are heated up to 65-80 °C, the production of a thermostable structure for phytase can be useful. In this study, we sought to perform bioinformatics analysis of the upstream region and protein structure of fungal phytase to improve its expression and thermostability properties. We used bioinformatics methods such as similarity search, multiple alignment, statistical analysis of physicochemical properties of amino acids, pattern recognition, and protein modeling to find out the effective factors in heat resistance of phytase. Change in Gibbs free energy (ΔG) of the best pattern promoter resulting from the interaction between RNA polymerase and the promoter sequences of modified genes of phytase was equal to -9 kcalmol-1, which is lower compared to other interactions. The evaluation of the three-dimensional structure of new phytases showed that amino acid substitutions aimed at improving thermostability did not change the form and structure of the protein. The results of Prochek, Whatcheck, and ERRAT for structural analysis and verification were 84, 72, and 70, respectively, that were satisfactory.
    Keywords: Fungal Phytase, bioinformatics, Homology Modelling, Molecular Docking, Upstream Region Analysis
  • F. Muhammad , , J. Hussain, S. K. Fareed, T. Ahmad Khan, S. Ahmad Khan, , A. Ahmad _ Pages 239-244
    Mycoplasma gallisepticum and Mycoplasma synoviae are the causative agents of avian mycoplasmosis in commercial poultry. Among the available tools, polymerase chain reaction (PCR) and culture are confirmatory tools for the diagnosis of mycoplasmosis after the initial serological screening of suspected birds. Overall, 181 samples were analyzed, 152 (84%) and 103 (57%) of which were found positive by PCR and culture, respectively. Further, 54 (92%) broiler samples were found positive for general avian mycoplasma. Among the total positive samples, MS positivity was as high as 72 (47%) by PCR, while it was 45 (44%) by culture. MG positivity was 23% and 25% in PCR- and culture-positive samples. MG grows more easily compared to MS. The agreement value between the tests was 67%. Overall, flock wise prevalence was not much varied. The prevalence of mycoplasmosis was higher during winter. Our study confirmed that PCR is the most sensitive and reliable tool for the diagnosis of avian mycoplasmosis in field samples.
    Keywords: Avian Mycoplasmosis, Mycoplasma gallispticum, Mycoplasma synoviae, PCR, Culture