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Wild and domestic ruminants become infected with Mycobacterium avium subsp. paratuberculosis (MAP) leads to chronic enteritis, known as Johne’s disease (JD). nA eight-year-old male wild goat (Capra aegagrus) that lived in the wildlife breeding center in Chaharmahal and Bakhtiari province with symptoms of prolonged diarrhea that did not respond to antibiotic treatment was referred to a veterinary center. After a while, the animal died. JD was diagnosed after laboratory diagnosis (by acid-fast staining). The present study reported the incidence of JD in a wild goat in Iran for the first time. The economic losses and animal health should be considered. Since 2020, the wild goat (Capra aegagrus) has been classified in the category Near Threatened (NT) near treated on the list (International Union for Conservation of Nature) IUCN. Therefore, protecting the health of these species is essential. The importance of carrier animals and reservoirs in wildlife should be given special attention.

Background and Objectives: Several studies have been carried out on the use of cell lines in researches, production and processing of drugs and biological products, and on the identification of toxicity and efficacy.The present study was conducted to determine the characteristics of BHK-21 cell lines as a suitable substrate for use in vaccine production and quality control, viral culture and research and diagnostic studies. Materials and Methods: In this study cloning of the cells by limit dilution was performed by preparing the appropriate seed cells. After checking the cell growth characteristics, sterility tests performed to ensure the absence of bacterial, fungal, mycoplasma, and mycobacterial infections. Detection of chromosomal diversity by karyotyping and cell identity using molecular tests along the CHO,LK,RBK,MRC5,Vero,GL cell lines was done. Also, master cell seed was tested for detection of adventitious viruses in two methods: in direct way through cytopathic effect during incubation of cell cultures and in indirect method using hemadsorption test, inculation into embryonic eggs, injection into mature and newborn mice. Results: The results indicated, this cell line had suitable growth charestristics without cross contaminations. Also the cell identity was confirmed by karyotyping and molecular tests. Conclusion:The results of various tests indicated that this cell line has the necessary requirements for use in the production and quality control of the biological products and viral researches.

Background and objectives: After using of trivalent oral poliovirus vaccine (OPV) for a long time in Iran, in recent years, due to the eradication of type 2 of poliovirus, a bivalent vaccine containing types 1 and 3 is used. Since it is a highly thermo labile vaccine strain of poliovirus, it should be stored under the recommended temperature. Because in vaccine storage and transportation, the cold chain may not be achieved in Iran tropical weather conditions, the potency of this vaccine may be subject to change. Methods: The purpose of this study was the stability evaluation of bivalent oral poliovirus vaccine produced by Razi Institute in Iran for stability study of vaccines to ensure constancy of virus at different temperatures and intervals freezing/defrosting cycles (stress tests). Three consecutive batches produced during full-scale production were sampled randomly. All quality control tests in the form of stability studies were done on vaccines stored at -20, 2-8, 22-25 and 35-37ºC in specific time intervals and 10, 20, 30, 40 and 50 freezing/defrosting cycles. Results: The findings indicate the stability of this vaccine in different situations. The results also showed the effects of the environment affecting factors on the potency of the vaccines and determined the correlation between the VVM grade and the vaccine potency. Conclusion: Important observations of the vaccine distribution network at different transit levels and the correlation of VVM grade and vaccine potency and freezing/defrosting are discussed in this study which will be of help to the vaccine program team.

Introduction: The prevalence of influenza epidemics and direct transmission of the emerged avian virus strains to humans have highlighted the necessity of improving the management programs against this disease. The aim of this study was to evaluate the biological adjuvant effect of hemokinin-1 (HK-1) on enhancing the immune response induced by the recombinant influenza vaccine based on the Nucleoprotein protein (NP) of the H1N1 influenza virus. Materials & Methods: In this experimental study, a total of 40 six-week-old female BALB/c mice were divided into two control groups (5 mice per group) and three treatment groups (10 mice per group). The control groups received normal saline and HK-1 oligonucleotide adjuvant. On the other hand, the treatment groups received NP recombinant protein (twice), NP protein with HK-1 (once), as well as NP protein with HK-1 (twice). Based on the grouping, 0.1 ml of each sample was injected into the quadriceps femoris muscle of the mice. The treatment groups received a 1:10 (v/v) ratio of the NP protein and HK-1. The serum samples were collected prior to primary and reminder vaccinations in defined intervals. The immune responses were analyzed using enzyme-linked immunosorbent and lymphocyte proliferation assays. Ethics code: RSVRI.REC.98.010 Findings: The rise of the anti-influenza antibody in immunized mice with recombinant NP-based vaccine received HK-1 adjuvant was significant, compared to that in the control group (P

Monoclonal Antibodies (mAbs) are used for biomedical research, diagnosis, treatment, production, and the quality control of biological products. mAbs are also very helpful in poliovirus research studies because it is still one of the major public health problems in developing countries. The main objective of this study was the production of mAbs against Poliovirus Type 2 (PV2) to be prepared and respond to the re-emergence of this virus. After fusion of immunized B cells prepared from mice with myeloma tumor cells and screening of about 250 hybridoma colonies, 22 with the highest antibody titer and without cross-reaction with others types were selected and cloned by limiting dilution. In the end, two colonies capable of secreting mAbs against epitopes of PV2 were used to produce mAbs. The mAbs were characterized by antibody assays, isotyping, and epitopes analysis using western blotting test, the cross-reactivity with other types, as well as stability, sterility, and mycoplasma tests. The results indicated that the produced mAbs had high specificity, sensitivity and stability against PV2 without any cross-reactivity and were of IgG1 Kappa chain with similar bands at 26 kDa during electrophoresis associated with viral protein VP3 neutralization. These mAbs were specific in serum neutralization tests for PV2 vaccine strain, and therefore, they are potentially valuable for routine polio research, diagnosis, isolation, production, and control of poliovirus vaccines.

ABSTRACT Background and Objectives: Poliomyelitis remains a major public health problem in developing countries, which signify the need for extensive diagnostic and prevention research. The aim of the present study was to design monoclonal antibodies (MAbs) against poliovirus type I with biomedical, diagnostic and therapeutic applications. Methods: B-cells were isolated from a mouse challenged with polio antigen injection. The B-cell were fused with myeloma tumor cells. After evaluation and screening of approximately 250 hybridoma colons by ELISA, 35 colons with the highest antibody titer and no cross-reactivity were selected and subsequently cloned by limiting dilution. Finally, three colons capable of secreting MAbs against epitopes of poliovirus type I were used for MAb production. Next, the MAbs were characterized by antibody assays, isotyping, epitope analysis (western blot), cross-reactivity test, stability test, sterility test and mycoplasma test. Results: The results indicated that the MAbs were of IgG1 kappa chain, had good stability and no cross-reactivity. In western blot, a band at 26 kDa which is associated to VP3 neutralization protein was observed. Conclusion: These serotype-specific MAbs can be potentially used for identification of type I poliovirus for research, diagnostic and prevention purposes. Keywords: Monoclonal antibody, Hybridoma, Poliomyelitis, Poliovirus.

Background: Problems of live and inactivated influenza vaccines such as, increasing emerge and re-emerge viruses with high human mortality, current epidemics of influenza and direct transmission of avian viruses to human, affect the vaccination program. DNA vaccines as third generation of vaccines is specially considered for control of influenza in human and poultry. The main advantage of these vaccines is humoral and cellular immune responses and broad spectrum of using these vaccines for control of circulating strains of influenza. In this study the conserved fragment of HA2 to form of DNA vaccine was designed to induce immunity against influenza viruses and its heterologous protective immunity against these viruses was evaluated. Methods: The experimental study was performed in Razi Vaccine and Serum Research Institute from December 2014 to July 2015 in Iran. The HA2 was cloned into pcDNA3.1 to assess the HA2 DNA vaccine and mice were immunized with the generated constructs in a DNA prime-DNA boost regimen in 4 groups. The humoral immune responses were analyzed at defined intervals by VN tests. The safety of the vaccine was evaluated by daily inspection and histopathological examination. For evaluation of cellular immunity, proliferation assay was used. Results: The antibody titre and cellular immunity of immunized mice was significantly higher than control group for two serotypes and the highest responses was in the group with two-time boosting (P

Purpose: The direct transmission of avian influenza viruses to human and increasing drug resisted strains posing new threats for public health. Therefore, development of efficient vaccines is needed to generate protective and persistent immunity to the viruses. Methods: Three motifs of Mx protein sequence in human, mouse and poultry located in interferon induced (GTP ase) domain were candidate as biologic adjuvant for enhancing the immune responses against influenza virus. Chimera proteins composed with the conserved HA2 subunit of influenza virus and the Mx motifs named HA2/Mx were modeled and evaluated by in silico analysis includes bioinformatics algorithms in order to explore biological characteristics of these peptides. Results: Amongst the predicted models, HA2/Mx1 peptide showed the better results following protein structures prediction, antigenic epitopes determination and model quality evaluation. Comparative homology modeling was performed with Swiss Model and the model was validated using ProSA. Epitope predictions revealed the construct could induce both B and T cell epitopes that expect a high immune response. Conclusion: Taken together, these data indicate that the HA2/Mx1 chimera peptide can be potentiated for developing an adjuvant-fused influenza vaccine capable of stimulating effective immune response.

Background: In the last decade due to emerge and remerge of influenza viruses, quality improvement of vaccines to increase immune responses in target populations have been more necessary. The potential of biologic adjuvant to stimulate and induce immune system is the basis of modern researches in prevention and controlling program of infectious diseases. In this study, the effect of the coding sequence of cellular Myxovirus resistance (Mx) protein as a biological adjuvant for inducing humoral immune response against influenza virus was investigated. Methods: The experimental study was performed on Balb/c mice in Razi Vaccine and Serum Research Institute from June to November 2014. Three conserved motifs of Mx were selected following sequence alignment between human, mouse and bird species. Potential of the motifs for stimulation immune responses against influenza virus were evaluated using in silico analysis. Based on the immune informatics data Mx1 sequence was the best immune inducer and cloned into pcDNA3.1 vector. Then formulated with inactivated H9N2 influenza antigen as adjuvant and injected to mice groups. The sera of vaccinated mice were collected prior to priming and boosting injections and also at defined weeks and analyzed with serological assays. Histopathological examination was done for evaluation of the vaccine and adjuvant safety. Results: The mean weight of the Balb/c mice in all control and treatment groups was similar and ranged from 21 to 37 gr (P= 0.05). The difference in increasing antibody titers against influenza virus in immunized mice who received Mx1-adjuvanted vaccine especially in second boosting was significant (P= 0.01) compared to the vaccine alone group. More than 78% of the immunized mice receiving two-time boosting have the mean antibody titer of >6 (Log2) which was higher (P= 0.001) comparing to the mice with one booster injection. Conclusion: These data revealed that Mx1 as biological adjuvant was able to increase antibody titer and induction memory immune responses against influenza immunization without causing any side effects.

Abstract With the combination of 3D printing and electroplating technique, metal-coated resin lattice is a viable way to achieve lightweight design with desirable responses. However, due to high structural complexity, mechanical analysis of the macroscopic lattice structure demands high experimental or numerical costs. To efficiently investigate the mechanical behaviors of such structure, in this paper a multiscale numerical method is proposed to study the effective properties of the metal-coated Body-Centered-Cubic (BCC) lattices. Unlike studies of a similar kind in which the effective parameters can be predicted from a single unit cell model, it is noticed that the size effect of representative volume element (RVE) is severe and an insensitive prediction can be only obtained from models containing multiple-unit-cells. To this end, the paper determines the minimum number of unit cells in single RVE. Based on the proposed method that is validated through the experimental comparison, parametric studies are conducted to estimate the impact of strut diameter and coating film thickness on structural responses. It is shown that the increase of volume fraction may improve the elastic modulus and specific modulus remarkably. In contrast, the increase of thickness of coating film only leads to monotonously increased elastic modulus. For this reason, there should be an optimal coating film thickness for the specific modulus of the lattice structure. This work provides an effective method for evaluating structural mechanical properties via the mesoscopic model.

Noise mitigation by means of the acoustic black hole (ABH) effect is a well-known engineering solution. However, the conventional method of applying ABH effect which requires modification of the structure geometry has various limitations which encourage the research of virtual ABH concept. In this study, the effect of ABH was applied through introducing virtual stiffness by a shunt circuit. According to the force-voltage electric analogy, stiffness has an inverse relationship with capacitance. So that the ABH effect can be virtually realized by following a power law profile using an array of independent capacitive shunts. The concept is studied through finite element simulation developing a macro code in ANSYS Parametric Design Language (APDL). To evaluate the influence of capacitance profile on the acoustic radiated power, parametric studies are conducted. Based on the results of the parametric studies, the capacitance profile is tuned for minimum radiated power. It is revealed that the virtual acoustic black hole (ABH) effect can offer 10.29%, 6.37%, and 7.47% reduction in the radiated power from the first to the last targeted mode, respectively. The virtual ABH effect introduced in this study can be used for semi-active structural noise isolation without any weight or manufacturing penalty.