Serum Research Institute Research Articles

Evaluation of the Razi Ornithobacterium rhinotracheale Vaccine by Experimental Challenge System Using LaSota Strain

BACKGROUND Ornithobacterium rhinotracheale (ORT) is a remarkable pathogen in the world poultry industry. The vaccine against this agent is used in poultry farms to prevent infection and reduce the incidence of disease. OBJECTIVES: In the present study, the efficacy of the first Iranian inactivated ORT vaccine produced by the Razi Vac-cine and Serum Research Institute was evaluated using the experimental challenge system . METHODS: Ninety day-old specific-pathogen-free White leghorn chickens were divided randomly into five groups of 18 chickens. The birds were housed in separate specific cages in isolation rooms. At the age of 14 days, the birds of two groups were vaccinated. Afterwards, at the age of 42 days, two groups of unvaccinated chickens and all of the vaccinated subjects were challenged with the LaSota strain of Newcastle Disease Virus (NDV) and ORT. One group of unvaccinated birds was maintained as the negative control. Blood samples were taken from chickens on days 14 (before vaccination) and 42 (before challenge) of the experiment. In addition, blood samples were collected on days 2, 4, 6, 8, 10, and 12 after the challenge (AC). On days 2, 4, 6, 8, 10, and 12 after challenging with ORT, the isolation and molecular detection of the bacteria were performed on samples from the trachea, lungs, air sacs, liver, and spleen. RESULTS: Following vaccination with the Razi ORT vaccine, the titers of antibody in vaccinated chickens were shown to be significantly higher than those of unvaccinated birds. In vaccinated groups, the ORT was not recovered in cultures from lungs, trachea, and air sacs. In the unvaccinated birds challenged with ORT, bacteria were isolated from lungs, tra-chea, and air sacs. Using the polymerase chain reaction method, ORT was only detected from samples of lungs, trachea, and air sacs 2 days after challenge (DAC) in vaccinated groups. Meanwhile, ORT was detected in lungs, trachea, and air sacs until 4 days after challenge in unvaccinated birds. CONCLUSIONS: We concluded that the Razi ORT vaccine was effective in protecting layer chickens against infection with serotype A of the ORT.

Effects of Cyclosporine Therapy on Liver and Kidney Retrieval in Poisoned Male Rats by Mesobuthus eupeus Scorpion Venom

Background: Mesobuthus eupeus venom is a member of Buthidae family, which can enter the blood circulation exerting detrimental effects on body organs, such as the liver and kidney through inflammation. Cyclosporine, known as an anti-inflammatory drug, is used to treat many inflammation-associated diseases. Objectives: In this study, cyclosporine was selected to inhibit the scorpion toxin effects on rat organs. Methods: This experimental study was conducted in the Razi Vaccine and Serum Research Institute, Agricultural Research Education and Extension Organization, Karaj, Iran, from June to November 2019. Fifty male rats were randomly divided into five groups of 10, including the control (10 mg/kg olive oil i.p), M. eupeus venom (10 mg/kg i.p.), cyclosporine 10 mg/kg (venom 10 mg/kg for 30 min i.p followed by cyclosporine 10/kg mg for 7 day i.p.), cyclosporine 20 mg/kg (venom 10 mg/kg for 30 min i.p followed by cyclosporine 20 mg/kg for 7 day i.p.), and cyclosporine 30 mg/kg (venom 10 mg/kg for 30 min i.p followed by cyclosporine 30 mg/kg for 7 day i.p.). After treatment with cyclosporine, the liver and kidney function was analyzed by calculating some biochemical enzymes, including serum glutamate-pyruvate transaminase (SGPT), serum glutamic oxaloacetic transaminase (SGOT), nitric oxide (NO), interleukin-2 (IL-2), malondialdehyde (MDA), creatinine, and urea via ELISA and spectrophotometry. Then, to determine the rate of apoptosis in tissue, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling method was done. Results: At the end of the study, the results showed a significant elevation in SGPT (164.5 ± 10 vs. 126.2 ± 7, P < 0.0001), SGOT (190.37 ± 11 vs. 148 ± 10, P < 0.0001), NO (24.4 ± 1.17 vs. 17.4 ± 1.4, P = 0.02), and MDA (0.42 ± 0.05 vs. 0.22 ± 0.04, P < 0.0001) in the venom group compared with the control group. There were no significant differences in the urea, IL-2, and creatinine between the venom and control groups. However, the group receiving cyclosporine (30 mg/kg) showed a significant decline in SGPT (96.42 ± 5.7 vs. 164.5 ± 10, P < 0.0001), SGOT (144.57 ± 9.24 vs. 190.37 ± 11, P < 0.0001), urea (28.83 ± 1.32 vs. 38.83 ± 1.6, P = 0.00), creatinine (0.023 ± 0.01vs. 0.29 ± 0.005, P < 0.0001), and MDA (0.10 ± 0.01 vs. 0.42 ± 0.05, P < 0.0001), as well as increased apoptosis rate (P < 0.05), compared with the venom group. No significant difference was observed between the cyclosporine and venom groups in NO and IL-2. Conclusions: Cyclosporine at a dose of 30 mg was able to decrease inflammatory responses and induce apoptosis rate. Therefore, it could be a suitable drug for patients bitten by a scorpion sting.

Improvement in the Growth and α-toxin Production of Clostridium septicum by Magnesium Sulfate.

Clostridium septicum, the anaerobic toxigenic bacterium is the agent that causes dangerous disease in man and animals. There is a lethal toxin of the bacterium namely alpha toxin. The ɑ-toxin has hemolytic, necrotic and lethal activities. Today, Razi Vaccine and Serum Research Institute of Iran produced the C. septicum vaccine in the form of bacterin/toxoid. Because of some problems, the vaccine needs to improve on an industrial scale. The study is going to find an appropriate supplement to improve growth and ɑ-toxin production. Three strains of C. septicum (vaccine, NH1 and NH8 strains) were cultured in the basic vaccine media. Magnesium sulfate, Copper, Ferrous, yeast extract, and trace elements plus vitamins' solution were added to the basic vaccine media in different cultures. The effect of the ingredients on the growth was measured by a spectrophotometer and the α-toxin secretion was assayed by hemolysin test. Growth of the bacterium and α-toxin secretion were increased by Magnesium (80 mg/l) in NH8 and vaccine strains significantly. The black precipitate was difficult to dissolve in magnesium media that must be solved. Trace elements plus vitamins solution mildly influence on NH1strain growth and toxin secretion. Other supplements (Cu, Fe, yeast extract) were not showen any significant changes in the growth and α-toxin production of C. septicum. Overflowing peptone (4%) in the vaccine media, fixes essentials of proteolysis activity, allows the sufficient growth and toxin production without Cu, Fe, and yeast extract. Due to essentially of Mg for growth, extra magnesium was added for improvement of media culture. The study suggests for Magnesium addition in the C. septicum vaccine media during production procedure after precipitation solving problem.

Immunogenicity and Efficacy of Live Infectious Bronchitis 793/B.08IR Vaccine in SPF Chickens

Infectious bronchitis virus (IBV) has a variety of serotypes with relatively limited cross-protection leading the disease to be a major problem in the poultry industry. The IBV 793/B strain has identified to circulate in Iran; therefore, the development of a specific vaccine to protect against the virulent virus has received attention. In this regard, the live IB 793/B vaccine (793/B.08IR) was developed in the Razi Vaccine and Serum Research Institute. In this study, the immunogenicity of 793/B.08IR vaccine via different routes of vaccination and efficacy of the vaccine were determined in specific-pathogen-free (SPF) chickens. Three treatment groups of 10 SPF chickens received the vaccine via eye drops, spray, and drinking water. The sera were collected from the chicks at 3 and 6 weeks after the vaccination, and IBV specific antibody was measured using enzyme-linked immunosorbent assay (ELISA) and serum neutralization (SN) test. To evaluate 793/B.08IR vaccine efficacy, 10 SPF chickens were vaccinated using eye drops. Moreover, 10 unvaccinated chickens were separately retained as negative controls. The birds were challenged with the virulent virus 3 weeks following the vaccination. Five days after the challenge, the tracheal swab was taken for virus reisolation. In the immunogenicity test, the ELISA titers of three vaccinated groups were significantly higher than the background values obtained in the control group (p<0.0001). The mean value of ELISA titer in the spray vaccinated group was higher than the spray and drinking water vaccinated groups 3 weeks following the vaccination; however, the difference was not statistically significant. No differences were observed in antibody titers among the three vaccinated groups 6 weeks after the vaccination. The results of the SN test confirmed the data obtained from the ELISA. The results of antibody titer and its increasing trend in chickens showed that 793/B.08IR vaccine induce proper immunity against the virus. In the efficacy test, IBV was isolated from 90% of the unvaccinated controls and 10% of vaccinated groups. The results of the recovery of the virus after the challenge showed that 793/B.08IR vaccine can provide a significantly improved protection against the pathogen in SPF vaccinated chickens.

Improvements to the live-attenuated Newcastle disease virus vaccine using Carbopol® 940 as a stabilizer.

Background and Aim:One strategy that can be used to stabilize vaccines is to convert them into a dry powder. This can protect the integrity of the active ingredients as well as vaccine antigenicity during manufacture, storage, and transport. This study highlights the potent adjuvant activity of Carbopol® when used alone to stabilize live-attenuated Newcastle disease virus (NDV) vaccines or when used in a formulation together with skimmed milk. Tolerability and potency of these formulations were compared with those obtained from other local live NDV vaccines produced locally by the Veterinary Serum and Vaccine Research Institute.Materials and Methods:We evaluated the cellular and humoral immune responses to a locally prepared, live-attenuated LaSota virus vaccine. Vaccine formulations were stabilized with Carbopol® 940 alone or in combination with skimmed milk.Results:Our results indicate that the use of Carbopol® 940 alone to stabilize a live-attenuated LaSota vaccine resulted in enhanced cellular and humoral immunity. The antibody titer was prolonged through the 6th week post-vaccination (5.0 log2). Full (100%) protection was observed in response to challenge with very virulent NDV at day 21 after vaccination; there were no clinical signs or lesions on examination. Addition of Carbopol® 940 to the live-attenuated vaccine formulation resulted in a more compact, stable, and high-quality lyophilized cake after freeze-dried lyophilization compared with that produced by stabilization with skimmed milk alone.Conclusion:Our data suggest that Carbopol® 940 may improve clinical responses to live-attenuated vaccines.

Molecular characterization of the 2018 outbreak of lumpy skin disease in cattle in Upper Egypt.

Background and Aim:Lumpy skin disease (LSD), an infectious disease of cattle, is characterized by raised nodules on the skin. Although the morbidity rate of LSD is low, it has a considerable fatality rate. Despite the annual mass vaccination of livestock with sheep pox vaccine (Veterinary Serum and Vaccine Research Institute, Egypt) enforced by Egyptian authorities, the LSD virus (LSDV) continues to circulate almost every summer. The present study aimed to discover the cause of cows naturally infected with LSDV circulating in Upper Egypt during the summer of 2018 using polymerase chain reaction (PCR) assay and to analyze their phylogenetics against reference genome sequences.Materials and Methods:We cultured LSDV in specific pathogen-free embryonated chicken eggs (SPF-ECE) and used conventional PCR to identify fusion and P32 genes, previously deposited in GenBank (MN694826, MN694827, and MN954664). Sequencing and phylogenetic analyses were performed on these two highly conserved viral genes.Results:LSDV infection of SPF-ECE resulted in characteristic white pock lesions. PCR products were identified on 1.5% agarose gel after electrophoresis at the expected positions for the fusion and P32 genes at 472 and 587 bp, respectively.Conclusion:The present study revealed that the two viral genes were identified from the Beni Suef and Sohag Governorates in all clinical cases and confirmed the circulation of LSDV in this outbreak. After sequencing, these genes were identical to those of the LSDV that had been identified and recorded in GenBank for the past 3 years.

Bradykinin-Potentiating Factors of Venom from Iranian Medically Important Scorpions.

The venom of animals, including snakes, scorpions, and spiders is a complex combination of proteins, peptides, and other biomolecules as well as some minerals. Among the biomolecules of some animal’s venom, small peptides that lack disulfide bands known as Non-Disulfide Bridge Peptides (NDBPs) potentiate the bradykinin by preventing the conversion of angiotensin 1 to angiotensin 2 using the mechanism of inhibiting the Angiotensin-Converting Enzyme activity and finally reducing the blood pressure in the victims. This feature of the NDBPs of animal’s venom is suggested as the potential of biological drugs. This study aimed to isolate venom components of three species of Iranian medically important scorpions and study the bradykinin potentiating effect of them. The scorpion specimens were collected from the venomous animals and antivenom production department of Razi Vaccine and Serum Research Institute, Karaj, Iran. Moreover, venom extraction was performed by electrical shock (5 volts). The obtained liquid venom of three species specimens was frozen and lyophilized immediately and then preserved in a cool and dried place. The isolation of the venom components for each scorpion was carried out using high-performance liquid chromatography. The obtained ranges of venom fractions (zones) were tested on isolated tissues of guinea-pig ileum and rat uterus using organ bath instrumentation in several replicates. The bioassays resulted in the peptides, including Z1 and Z2 regions in the venom fractionsof the Hottentotta saulcyi, Z2 in Odontobuthus doriae, as well as Z2 and Z3 in Mesobuthus eupeus demonstrated bradykinin potentiating effect. It is concluded that Bradykinin Potentiating Factors were traceable in the venom of all three scorpion species. Therefore, these venoms have the therapeutic potential to exploit biological-based drugs.

Monoclonal Antibody Preparation Against Nucleoprotein of Avian Influenza Virus Subtype H9N2

Background: In recent years, various poultry diseases have posed risks to this industry. Respiratory and infectious diseases are the most common diseases. According to previous findings, influenza disease is recognized as a significant life-threatening disease in the industry of poultry worldwide. Influenza virus type A, which belongs to the family Orthomyxoviridae, is responsible for a serious infectious disease. H9N2 AVI subtype circulates in the poultry worldwide, causing significant economic losses and infections in humans, also domestic and wild animals. Objectives: The purpose of the present experimental research was monoclonal antibodies (MAbs) production in contradiction of the NP of avian influenza virus (AIV) H9N2 subtype to detect AIV antigens and antibodies in the Department of Proteomics and Biochemistry, Razi Vaccine and Serum Research Institute, Agricultural Research Education and Extension Organization (AREEO), Karaj, Iran. in 2017. Methods: The conserved protein of NP from H9N2A/Chicken/Iran/259/2014 virus, with 60 kDa molecular weight, was isolated using the electroelution method. The purified protein was applied for Australian BALB/c mice immunization. After evaluating immunization by ELISA assay, the spleen of immunized mice was isolated and hybridized with SP2/0 myeloma cell. Next, the hybridized cell was cultured, and clone soups were collected after 15 days to examine antibodies via ELISA assay. The produced antibodies using Western blotting and antibody isotype kits were characterized. Results: Thirty antibody-producing clones were examined for reactivity against Nucleoprotein (NP), the antigen, at 1 µg/mL concentration. According to the ELISA assay of antibody titers, two (3/F10 and 2/D7) out of 30 antibodies were bound to the antigen with titer 0.863 and 1.641, respectively. Two hybrid clones, 2D7 and 3F10, which produced anti-NP antibodies, were isolated and cultured. Characterizing of the produced antibodies using Western blotting was performed using H9N2 virus; finally, two clone soups (3/F10 and 2/D7) reacted with the NP virus protein. According to the isotyping of antibodies produced by 3F10 and 2D7 clones, 3F10 clone produced IgG1 with a κ chain, and IgG1 concentration was 1.997, based on ELISA assay. Also, 2D7 clone produced IgG2a with a κ chain, and IgG2a concentration was obtained 1.951. Conclusions: According to the findings of our study, the produced antibody might be used in the diagnosis of influenza.

Purification of Nucleoprotein of H9N2 Avian Influenza Virus Strain by Electroelution

Background: Avian influenza virus (AIV) belongs to the family of Orthomyxoviruses type A and causes avian influenza (AI) infectious disease. Currently, serological diagnostic techniques such as agar gel propagation (AGP), hemagglutination inhibition, and enzyme-linked immunosorbent assay (ELISA) are considered as important tools for the antibodies detection against viral antigens. Due to antigenic variation in the surface of AIV glycoproteins (hemagglutinin and neuraminidase), these proteins cannot be used in serological tests. Development of assays to detect AI surface glycoproteins is problematic because a great variety of combinations of these subtypes are found in nature. The internal antigen determinants on the nucleoprotein (NP) are highly conserved within influenza viruses, making this protein more appropriate for a serological test. Objectives: In the experimental present study, an effectual method was expanded to purify NP of H9N2 AIV based on Electroelution method. Methods: AIV strain A/flash chicken/Iran/772/1998 (H9N2) was acquired from the Department of Avian Influenza Reference Laboratory, Razi Vaccine and Serum Research Institute, Iran, about 2 cc, in 2017. Nucleoprotein of AIV (H9N2) was purified by an efficient and simple modified method directly from native polyacrylamide gel electrophoresis (PAGE) according to the Electroelution method. The purified protein concentration was defined by the Lowry method, and the purified NP protein (60 KDa) was examined by Tricine-SDS-PAGE. Results: The protein concentration of the virus solution was 4.62 mg/mL by the Lowry method. The purified Nucleoprotein concentration was 0.296 mg/mL by Lowry method and the Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis results showed only a 60-KDa protein band in the gel. Conclusions: The current technique was simple and rapid and made it possible to isolate NP from the H9N2 virus. The Nucleoprotein antigen is an appropriate candidate potential to detect antibodies against all subtypes of AIV and used as the main target antigen for the diagnosis of influenza virus due to its very high scale of sequence preserved among exist strains.

A Case of Identity Confirmation of Brucella abortus S99 by Phage Typing and PCR Methods.

Brucellosis is a zoonotic infection that is associated with fever in humans and abortion in animals. The agent of this disease is a facultative intracellular gram-negative coccobacillus called Brucella. There are six classic species, including B. abortus, B. melitensis, B. suis, B. canis, B. neotomae, and B. ovis. In recent years, four new species have been reported, including Brucella ceti, B. microti, B. pinnipedialis, and B. inopinata. Human disease causes hygienic and economic losses, including inactivity of workforces in the community and high cost of treatment. The disease also causes catastrophic losses in the livestock industry. There is no effective vaccine against human brucellosis. Hence, attempts to prevent human infection with Brucella are focused on preventative measures, including control of infection in livestock, which lead to a reduction in its incidence in humans. The common methods for diagnosis of this disease are serologic methods including Rose Bengal, Wright -2 ME and the ring test. B. abortus strain S99 is used to produce these diagnostic antigens. The production of these antigens requires the presence of a well-characterized seed with full identity. The aim of this work was confirmation of the identity of B. abortus S99 by phage typing, AMOS and multiplex PCR techniques. Therefore, it is essential to carry out the identification of the strains used as seed for the production of the brucellosis diagnostic antigens. In this project, B. abortus strain 99 was supplied by the bacterial collection of the Brucellosis Department of Razi Vaccine and Serum Research Institute. Then, the main aim of the present study was the confirmation of the seed identity by doing the tests through the standard phage typing method, AMOS PCR and multiplex PCR (Brucladder) methods. Results were in support of the identity of the studied strain, and the molecular methods could also be used as the sensitive approaches for validation of antigenic seed.

Propagation, Purification And Molecular Characterization Of Rvf Virus (Zh 501) Strain For Vaccine Production In Egypt

Rift Valley Fever (RVF) is an endemic disease in Egypt causing disease in animals and humans since the 1977. Full molecular description of the national vaccine strain is a strategic concern to ensure the antigenic makeup of local vaccine is adequate to provide the required protective immunity for the vaccinated animal species and as a tool of final product quality control. Molecular characterization and phylogenetic analysis of Gc gene of M segment of RVF-ZH501 strain which used in local vaccine production in veterinary serum and vaccine research institute (VSVRI) were carried out in this study. RVF-ZH501 strain was propagated by passages on BHK cell line and then purified my plaque purification technique. RT-PCR for amplification of the Gc gene of M segment. Sequence analysis of the obtained PCR product carried out by MEGA7 program. The result of the study revealed that RVF strain ZH501 Gc genomic content has 99.5 % molecular identity to the strains firstly isolated in Egypt in 1977, so still suitable for vaccine production.

Genetic analysis of clinical and vaccine strains of Bordetella pertussis by Pulsed-Field Gel Electrophoresis (PFGE), Multi Locus Sequence Typing (MLST) and serotyping

In spite of high vaccination coverage in the Expanded Program of Immunization (EPI), pertussis has not been eradicated yet and the re-emergence of the disease is still reported worldwide. The genetic divergence study of circulating clinical strains of Bordetella pertussis among the population with high vaccination coverage is a useful tool to have an insight in the understanding of genetic patterns of this bacterium and deviation of them from vaccine strains. Different methods are accessible for studying of Bordetella pertussis that can perform appropriate assessment between populations. Strains used in this study were a collection of two pertussis vaccine strains used to create killed pertussis vaccine over years at Razi Vaccine and Serum Research Institute, 10 clinical and 2 reference strains (ATCC9797 and Tohama I) in Multilocus Sequence Typing (MLST), Pulsed-Field Gel Electrophoresis (PFGE), and serotyping. The genetic profiles of vaccine working and master seeds showed no important change(s) in frequencies of fingerprint types investigated in the vaccine strains and had homogeneity in PFGE method where the clinical isolates showed diversity in genetic profile. Serotyping method showed that all of 10 clinical strains expressing Fim 3. In MLST study, seven housekeeping genes including adk, pgm, fum C, tyr B, gly A, pep A and icd were analyzed which showed no changes in the sequence of clinical and vaccine strains with 100% homology. The genes that cause pathogenicity like ptxC, tcfA and fhaB were also evaluated and the results illustrated heterogeneity in the vaccine and circulating strains.

Evaluation of Pertussis Toxin Expression in B2 and THIJS Media.

Whole-cell pertussis vaccine (wP) has been imperative and highly effective in preventing childhood deaths due to pertussis. Pertussis toxin is one of the virulence factors of Bordetella pertussis in all available pertussis vaccines. wP production in Razi Vaccine and Serum Research Institute is according to bioreactor culture of B. pertussis strains in B2 medium. The aim of this study was to evaluate B. pertussis strain 509 PT production in B2 and Thalen-IJssel (THIJS) media by Chinese Hamster Ovary (CHO) cell and enzyme-linked immunosorbent assay methods (ELISA). In the current study, B. pertussis strain 509 was cultured in B2 and THIJS media. Six samples were taken during the log growth phase within 2-3 h intervals (triplicate). The growth rate was calculated using opacity and the quantification of cell-associated and released PT measured by ELISA and CHO cell assays. THIJS medium was significantly showed an increase in the bacterial growth rate. During the first 29 h, bacterial concentrations in B2 and THIJS culture medium were 19 and 29 IOU, respectively. In THIJS medium, greater amount of pertussistoxin production was cell-associated. In B2 medium, maximum cell-associated toxin by ELISA and CHO cell assays were in the ODs of 1.1 and 0.9 and for THIJS medium in the ODs of 1.6 and 1.1, respectively. B. pertussis strain 509 in THIJS medium produced higher cell mass and cell-associated pertussis toxin than that of B2. It can be used for the production of whole-cell vaccine with higher pertussis toxin and accordingly using lower biomass per dose leading to the reduction of vaccine toxicity.

A very French connection, a brief discourse in commemoration of life and services of the late Louis Pierre Joseph Delpy on his 120th birthday anniversary.

Razi Vaccine and Serum Research Institute (RVSRI) turned 95 years old in 2015. Majority of the animal infectious diseases such as rinderpest and anthrax that used to frequently strike the historic Persia are now gone for good or under control owing to the pioneering researches conducted at the institute in the early-mid 20th century in the field of vaccine manufacturing. The earliest such scientific contributions, were truly made by the French eminent veterinarian Dr. Louis Pierre Joseph Delpy who joined the institute in 1931. In his 18 year-long directorship tenure he taught his colleagues fundamentals of vaccinology, basics of modern epidemiology, essentials of infectious disease control disciplines, the art of scientific writing and much more things that changed the institute for ever. This paper reviews the events and turning points in the first 25 years of service of the institute in a chronological way and remarks Delpy’s principle involvements in all of these on the occasion of the 120 anniversary of his birth. At the entrance of the institute headquarter building where his bronze bust is placed, visitors can see a memorial etched plate that reads “... The architect of Razi and founder of Archives De L’Institute Razi (Archives of Razi Institute) was an enthusiastic scientist with a creative mind. … For the Razi community, Dr Delpy is gone but not forgotten.”

Molecular Identification of Six Honeybee Viruses in Iranian Apiaries.

The identification of honeybee viruses is of serious importance, particularly considering the lack of information on the natural incidence of viral infections in honeybee populations worldwide. Moreover, the global spread of Varroa destructor in honeybee colonies has a significant effect on the viral infection. In the present study, 160 samples of adult bee from apparently healthy colonies but with a background of parasitic diseases, tremor, and paralysis, were collected during 2011-2012. The samples belonged to 23 different provinces of Iran. They were sent to Razi Vaccine and Serum Research Institute, Karaj, Iran, for further analysis, and examined for the presence of viruses using reverse transcription polymerase chain reaction assay. According to the results, out of 160 samples, 9 (5.8 %), 40 (25.6 %), 12 (7.8 %), 34 (21.8 %), 7 (4.5 %), and 29 (18.5%) cases were positive for acute bee paralysis virus (ABPV), black queen cell virus (BQCV), chronic bee paralysis virus (CBPV), deformed wing virus (DWV), Kashmir bee virus (KBV), and sacbrood virus (SBV). The samples collected from 18 provinces (78 %) were positive for at least one virus. Among all samples, 83 (53.2 %) specimens were infected with at least one virus. The highest prevalent virus was BQCV, followed by DWV, SBV, CBPV, ABPV, and KBV, respectively.

Radiolabeling the Peptides extracted from Scorpion venom (ICD-85) with 99mTc as treatment and imaging agent for cancer

Introduction: Venoms are a mixture of proteins and peptides which can be used for fighting cancers. They can affect the exposed fibers and the muscles directly or through motor nerves, causing neuromuscular intoxication. ICD-85 is a combination of three poly-peptides derived from the venoms of an Iranian brown snake and a yellow scorpion. Materials and Methods: Technetium-99m was obtained from a 99Mo/99mTc generator. Venom (ICD-85) was provided by Razi Vaccine and Serum Research Institute of Iran. Labeling of this venom was achieved with Technetium-99m. Labeled ICD-85 was injected into mice to determine the organ targets and excretion pathway. Results: Results show that the radiolabeling yield of the radio-venom is acceptable. The biodistribution study shows that the organs of liver, kidney and tumor are the targets of ICD venom. Also moderate clearance of the complex from blood was seen. Conclusion: The results suggest that 99mTc radiolabeling of venom (ICD-85) may be a good tool for in-vivo studies and is an excellent approach to follow the process of biodistribution and kinetics of toxins for cancer imaging and treatment.

EVALUATION OF IMMUNOGENICITY OF DNA VACCINE CODING OUTER MEMBRANE PROTEIN 31 (OMP31) OF BRUCELLA MELITENSIS IN MICE.

28Feb 2017 EVALUATION OF IMMUNOGENICITY OF DNA VACCINE CODING OUTER MEMBRANE PROTEIN 31 (OMP31) OF BRUCELLA MELITENSIS IN MICE. Yasser M. Kamel , Noha A. Helmy , Amani A. Hafez and Safaa M. Barghash. Infectious Diseases Unit, Animal Health Department, Desert Research Center, Cairo, Egypt. Veterinary Serum and Vaccine Research Institute,Sera and Antigens Department, Abbasia, Cairo, Egypt. Parasitology Unit, Animal Health Department, Desert Research Center, Cairo, Egypt.

Correction to: Evaluation of Colicin Effect on the Induction of Treated Mice in Prevention of Infection Caused by Escherichia coli K99 [Published in Int J Enteric Pathog. 2016;4(4):e37106

This correction presents the amended version of affiliation number 1. 1Microbiology Department, Razi Vaccine and Serum Research Institute Shiraz Branch, Agricultural Research, Education and Extension Organization (AREEO), Shiraz, Iran

Development of Indirect Immunofluorescence Technique for the Identification of MRC5 Working Seed Cell.

Diploid and continuous cell lines are used to propagate viral vaccines. At Human Viral Vaccine Department of Razi Vaccine and Serum Research Institute, MRC5 diploid cell is used for the development of live attenuated measles, mumps, rubella, and three types of poliovirus vaccines. Additionally, three continuous cell lines (i.e., RK13, HeLa, and Vero) are applied in quality control tests. Accordingly, cell cross-contamination can occur at cell culture labs, hence controlling the identity and specificity of cells is essential. Indirect immunofluorescence is a sensitive, specific, and simple test for cell identification. The present study was designed to develop the in-house indirect immunofluorescence test (IIF) as follows: homemade polyclonal anti-MRC5 serum was prepared in rabbits, and cross-reactive antibodies to RK13, HeLa, and Vero cells were eliminated. The diploid and continuous cell lines were fixed on Teflon slide using cold methanol and acetone. The reproducibility of the in-house IIF test was evaluated using the agreement Kappa test. The purity of the three batches of MRC5 working seed cell at Human Viral Vaccine Department of Razi institute was verified using IIF and no contamination with continuous cell lines was detected.

Comparative Sensitivity Analysis Between two methods for Species Differentiation and Interspecies Cross Contamination in Animal Cell culture

Background and Aims : The present article focuses on design and development of a highly sensitive and convenient approach for rapid detection of animal species and cross contaminations quickly during cell cultures as most important document for manufacturing working cell bank system. This test is one of the four most important documents during implementing the banking system. By using this modified test also one of the major risks in cell culture laboratories, cross- contamination and misidentifications of cell lines will be also under test. Materials and Methods : A PCR _RFLP assay was optimized based on the use of a pair of primers that anneal to a portion the cytochrome b gene in all the species. The amplification product was digested with a panel of six restriction enzyme and the pattern derived was resolved on 3% high resolution agarose gel for 2 species, human & primate. As a control test we used from conventional method iso enzyme assay. Results : This protocol produced a unique restriction pattern and the origin of this animal cells resulted to be confirmed by this analysis. Its sensitivity in detecting interspecies cross contamination was at least 100 pg DNA/reaction, which was sufficient for detection of each species of DNA. Conclusion : For the last few years the RAZI vaccine and serum research institute in IRAN has been involved in developing such an ideal Iranian vaccine based on local Iranian material. The method developed in this study will provide a useful tool for the authentication of animal species comparable and most time consuming, cost benefit to other conventional analysis. Using the method (PCR-RFLP) developed during this research not only significant differences between human and non human also cross- contamination between different cell lines can simply distinguish.