نشریه تحقیقات دامپزشکی و فرآورده های بیولوژیک
سال سی و ششم شماره 4 (پیاپی 141، زمستان 1402)

Vaccination against bird flu H9N2/H5N8 is considered as an effective strategy to deal with and control these deadly pathogens. For this purpose, in this study, an effective dual chimeric vaccine against H9/H5 from the bidirectional HA2 region of H9N2 and H5N8 was designed and analyzed using bioinformatics models, physicochemical characteristics and antigenic structure analysis of selected epitopes.To carry out this study, (A/chicken/Iran/B308B/2004) H9N2 and (A/Poultry/Iran/clade 2344/2018) H5N8 strains were selected and the epitopes stimulating B and T lymphocytes were investigated and the most suitable ones were selected. Then a chimeric vaccine was designed by combining highly immunogenic epitopes. In the following, using related software, the physicochemical characteristics and secondary structure of the designed vaccine were evaluated in terms of thermal stability, hydrophilicity, isoelectric pH, antigenicity and allergenicity. Finally, the proposed vaccine was reverse transcribed and adapted to be placed in the prokaryotic expression vector pET-41 a(+). According to the results, it is predicted that the antigenic structure designed in this study can be expressed successfully in prokaryotic systems and use for immunogenic studies against H5N8 and H9N2 strains of influenza virus.

The aim of this study was to evaluate the sperm quality preservation in sheep using mitochondria-targeted antioxidant Mito-TEMPO during chilling process. In this experiment, semen samples were collected from five Zandi rams and added to the extender containing 0, 0.5, 1, 5, 50 and 500 µM Mito-TEMPO and stored at 4 ˚C until 48 hours. Sperm quality was evaluated at 0, 24 and 48 hours after chilling and total motility, progressive motility, viability, membrane integrity, mitochondrial activity and lipid peroxidation were assessed. No difference was observed between treatments at time 0 of cooling. During 24 and 48 hours cooling periods, using 5 and 50 µM Mito-TEMPO increased total motility, progressive motility, viability, membrane integrity, mitochondrial activity and decreased lipid peroxidation in ram chilled sperm. In conclusion, using 5 and 50 µM Mito-TEMPO in chilling herbal extender could be a suitable method to conserve ram sperm quality during chilling process.

Domestic pigeons are in close contact with humans and other birds in urban areas. As a carrier of Escherichia coli and Salmonella spp, these birds can play a significant role in spreading and transmitting these pathogens to humans and industrial poultry. The aim of this study was to isolate Escherichia coli and Salmonella spp from the cloacal of the domestic pigeons in Ardabil, northwest of Iran and determine their antibiotic resistance. For this purpose, 130 swab samples were prepared from healthy household pigeons in different parts of Ardabil city. After cultivation and isolation of E.coli  and Salmonella spp  in selected media, their colonies were identified by biochemical methods such as EMB, TSI, SIM, MR-VP, indole and urea. Antibiotic resistance of the isolates were determined according to the standard method of disc diffusion. Out of 130 samples, 23 cases (17.69%) of E.coli  and 7 cases (5.38%) of Salmonella spp were isolated. Antibiotic resistance test on E.coli showed that tetracycline and sulfadiazine + trimethoprim had the highest resistance with 86.95% and 82.60%, respectively. While enrofloxacin and gentamicin showed the lowest resistance with 21.73% and 17.39%, respectively. In Salmonella spp  isolates, the highest resistance to tetracycline (100%) and sulfadiazine + trimethoprim (85.71%) and the lowest resistance to co-amoxiclav (14.28%) and florphenicol (14.28%) were observed. All isolates of E.coli  and Salmonella spp were sensitive to ciprofloxacin. The results of the present study indicate the presence of E.coli and Salmonella spp contamination in Ardebil urban pigeons,and also have significant resistance to some antibiotics. The implementation of strict health programs and the principled use of antibiotics in pigeons seem necessary to prevent the transmission of contamination and the spread of antibiotic resistant strains to other poultry and human.

Probiotics are live and beneficial microorganisms that are known as a factor for prevention of infectious diseases and cancer. In this study, the effects of cytotoxicity of Lactobacillus plantarum extract on the colon cancer cell line (Caco-2) as well as the expression of VDR, IL3 and NOD2 genes were investigated.The supernatant of L. plantarum culture was collected. Then, the effects of cytotoxicity and fluidity of bacteria and Supernatant on Caco-2 cancer cell line were investigated using MTT method. Also, the expression of VDR, IL3 and NOD2 gene was evaluated using Real-Time PCR. Data were analyzed using one-way ANOVA. Investigation of the inhibitory effect of L. plantarum culture supernatant on Caco-2 cells and gene expression at different concentrations showed that 100 μL/mL for VDR, IL3 and NOD2 had the highest increase in gene expression significantly after 48 hours. The MTT test showed the highest lethality at a concentration of 100 μL/mL. Gene expression at 100 μL/mL increased significantly at 24, 48 and 72 h for VDR, 48 h for IL3 and 72 h for NOD2 compared to the control group (P <0.05). L. plantarum can be used to develop a new treatment strategy with high efficacy, low side effects, biologically safe and lower cost. Based on this, it is suggested to investigate the treatment and prevention of colon cancer using L. plantarum.

Turmeric extract has antioxidant, antibacterial, antifungal, antiviral, and anti-inflammatory effects, and in addition, curcumin present in turmeric extract can be a powerful protective factor against biochemical changes and cellular oxidative damages. In order to investigate the effect of turmeric aqueous extract on lipid peroxidation and the activity of antioxidant enzymes in the liver tissue of chick embryos treated with silver nanoparticles,  45 race Ross 308 fertilized eggs were randomly divided into control groups (single injection of 0.5 ml saline solution) and 4 treated groups. On the 10th day of incubation, treated groups of 1, 2, 3 and 4 received silver nanoparticles (a single injection of 0.5 ml silver nanoparticles, 200 ppm and 60 nm). On the 12th day of incubation, treated groups of 2, 3 and 4 received turmeric aqueous extract with concentrations of 100, 200 and 300 μg/ml (single injection of 0.5 ml turmeric aqueous extract). The injection was done in the amnion sac embryos. On the 20th day of incubation, the concentration of malondialdehyde and the activity of superoxide dismutase, glutathione peroxidase and catalase enzymes in liver tissue were measured by ELISA method. Statistical analysis was carried out by using one-way ANOVA and post-hoc Tukey tests (p<0.05). Compared to treated group 1, the tissue concentration of superoxide dismutase, glutathione peroxidase and catalase in treated groups 2, 3 and 4 significantly increased and the tissue concentration of malondialdehyde decreased significantly (p<0.05). Dose-dependent injection of turmeric aqueous extract dreduces the toxicity caused by silver nanoparticle and reduces oxidative stress and lipid peroxidation in liver tissue of chick embryos.

Due to the increase in the price of inputs and the necessity of nutrition management; the effect of food restriction on performance, growth and immune response was investigated in 960 Arian breed broiler chickens (480 male & 480 female). Therefore, two periods of food restriction were considered separately in two treatment groups (at the second week and the sixth week) using four diluted rations (0, 20, 30 and 40%) in three replications in a completely randomized design. After analyzing the data, the average traits were compared with Duncan's multiple range test, the effect of food restriction on the immune response to ND, AI, IB and IBD vaccines was also investigated with serology on the 20th and 42nd days of rearing, According to the results , the broilers in the first group with 20% dilution diet reached the weight of the control group (p < 0.05) at the age of 42 days, also the corrected food conversion coefficient was improved and had compensatory growth; In the second group, the chicks could not reach the control group at the age of 42 days (p < 0.05) and the improvement percentage index (weight gain) was not favorable, and more time was needed to reach the appropriate improvement index, and the same time the ability to compensate for the delay in growth and improvement. The response of the immune system to vaccination against ND, AI, IB and IBD in both treatment groups with low food restriction (20%), improved and significantly differen from the control group (p < 0.05), the titers of the first group treatments it was better than the second group  and it was lower in male than female chickens.

Sheeppox is one of the most common diseases in sheep that can spread quickly in the herd. Currently, there is no specific treatment for this viral disease. The purpose of this study was to investigate the antimicrobial peptide CLF36 and CLFarm on the surface protein of the sheep pox virus P32 protein and the cellular receptors of this protein such as heparin sulfate and UDP-N-acetyl glucosamine. The third structure of P32 was done using software such as I-TASSER. The interactions were performed statically in a computer by bioinformatics tools such as HDOCK software and using Gromacs in the molecular dynamics section. The results of this study showed that the amino acids of CLF36 peptide involved in hydrogen bonding with P32 include D1, K5, V8, Q11, K9, R16, R22, K25, R27 and K34 and for CLFarm include K5, K9, Q11, K13, R27, W30, Q31 and K35. Docking results showed that peptides were attached to three epitope regions EP1,4,8 in P32 protein. In connection with the molecular docking between CLF36 arm and the two receptors UDP-N-acetylglucosamine and heparin sulfate, the hydrogen bonds included Gln 11, Trp 30 and Trp 4, Trp 30, Gln 11, Arg 27, Cys 33, respectively; Although CLF36 had weak binding energy with these two receptors. In the molecular dynamics section, the graph of RMSF and RMSD and Gyrate showed a few fluctuations, which were ignored. Finally, these results showed that both peptides have a good performance against P32 in a period of 50 ns.

The spread of bird flu disease with high mortality, reduced egg production and forced killing of infected flocks has a devastating effect on the poultry industry. Based on the central protein, the virus is divided into three subgroups A, B, and C. Influenza type A virus is very common. This virus can infect different species of mammals and birds, and it is more pathogenic than influenza B and C, and it causes epidemics of different severities almost every year. Breeding goals to maintain the health of animals lead to the well-being of animals and prevent economic losses. The purpose of this research was to use biological network analysis method to identify new key markers involved in bird flu disease. In this study, in order to quickly obtain the most important genes involved in this disease, gene expression microarray data related to the lung tissue of chickens infected with the virus was prepared from the GEO site with the accession number GSE53931. The protein-protein interaction network was drawn by Cytoscape software. Protein complexes were analyzed using MCODE. Three protein complexes were extracted. The seed proteins in the complexes include RPL10A, NOB1, and MSH4, which were introduced as key markers, as well as NEB, RPF1, CSRNP1, RNF146, and RPL18A genes with the lowest level of expression and SLC6A20, PLN, SLC17A8 ,NOXO1 and TMEM179 gene with the highest level of expression in the present study. Most of the genes are involved in the transcription, replication, translation of the gene in the influenza virus. Network-based analyzes can detect effective genes and functional modules, which can significantly help in identifying regulatory factors and biological processes involved in the response to avian influenza, and subsequently, better control of the disease.

In present study, in order to evaluate scolicidal effect of CM11anti-microbial peptide (α-helical peptide with 11 amino acids) against hydatid cyst, Echinococcus granulosus protoscolices were collected and washed with normal saline. At first, the percentage of viability of protoscolices was evaluated using eosin staining. Different concentrations of CM11 peptide (16, 64, 128, and 256 μmol.), along with positive and negative control group, were used against protoscolices for 30, 60, 120 and 240 minutes. Eosin staining was done at the end of the incubation time, and the images were taken with a camera equipped with Tcapture software. Finally, the number of dead and live protoscolices was determined by ImageJ software, and the viability and mortality percent was calculated. Data analysis was carried out by using Sigma Stat (version 3.5). P value < 0.05 was considered statistically significant. Up to 2 hours, the effect of all concentrations of CM11 peptide on protoscolices was not significant compared to the negative control group. However, after 4 hours of exposure, 128 and 256 μm of peptide, were significantly decreased the viability of protoscolices (P<0.05). With regard to the results of this study and considering the resistance of the present antiprotoscolics drugs, the CM11 can be used as a relatively effective peptide against protoscolices in future research.

The present study was conducted to compare the effect of methylprednisolone administration by two intramuscular injection and oral methods on liver enzymes and hemodynamic factors. This study was a multi-group clinical trial conducted on 15 healthy domestic short-haired cats. The samples were randomly divided into 3 control groups (receive 5 cc of distilled water), oral prednisolone (5 mg/kg) and intramuscular injection prednisolone (5 mg/kg). Hemodynamic factors (heart rate, breathing and body temperature) before drug administration and after 10, 30 and 60 minutes and liver biochemical factors (ALP, AST and ALT) before drug administration and 24, 48 hours after that were measured. Changes in body temperature were not statistically significant in any of the groups. However, the changes in the respiratory rate in the oral prednisolone group (P=0.007) and the intramuscular injection group (P=0.04) were significantly difference. Changes in mean heart rate (P=0.007), ALT (P<0.005) and ALP (P=0.009) were significant difference in the intramuscular injection group. Moreover, AST level was significant difference in oral prednisolone group (P=0.003) and intramuscular injection group (P=0.04). The results of this study revealed that the administration of prednisolone at a dose of 5 mg/kg both orally and by intramuscular injection can lead to changes in the level of liver enzymes, heart and respiratory rate in the short term. The amount of these changes was higher in intramuscular injection method in compare to oral method. Therefore, in order to use this medicine, it is better to use the oral form, although the patient's vital factors must be carefully checked.