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The discrimination power of Salmonella molecular typing by combination of ERIC, REP and BOXAIR methods, or by combination of BOXAIR with ERIC or REP, is sufficient to determine genetic relationships for epidemiological purposes.

Background Green synthesis of nanoparticles by plant extracts plays a significant role in different applications. Recently, several studies were conducted on the use of nanoparticles as adjuvant. The main aim of this study was to evaluate green synthesized silver nanoparticles (AgNPs) as adjuvant in rabies veterinary vaccine and compare the results with the existing commercially available alum adjuvant. Materials and methods In the current study, AgNPs were prepared by the reduction of aqueous silver nitrate by leaf extract of Eucalyptus procera . The formation of AgNPs was confirmed by ultraviolet (UV)–visible spectrophotometer, scanning electron microscopy, dynamic light scattering, and X-ray diffraction analysis. Then, different amounts of AgNPs (200 µg, 400 µg, 600 µg, and 800 µg) were added to 1 mL of inactivated rabies virus. The loaded vaccines (0.5 mL) were injected intraperitoneally into six Naval Medical Research Institute mice in each group on days 1 and 7. On the 15th day, the mice were intracerebrally challenged with 0.03 mL of challenge rabies virus (challenge virus strain-11, 20 lethal dose [20 LD 50 ]), and after the latency period of rabies disease in mice (5 days), the mice were monitored for 21 days. Neutralizing antibodies against rabies virus were also investigated using the rapid fluorescent focus inhibition test method. The National Institutes of Health test was performed to determine the potency of optimum concentration of AgNPs as adjuvant. In vitro toxicity of AgNPs was assessed in L929 cell line using MTT assay. In addition, in vivo toxicity of AgNPs and AgNPs-loaded vaccine was investigated according to the European Pharmacopeia 8.0. Results AgNPs were successfully synthesized, and the identity was confirmed by UV–visible spectrophotometry and X-ray diffraction analysis. The prepared AgNPs were spherical in shape, with an average size of 60 nm and a negative zeta potential of −14 mV as determined by dynamic light scattering technique. The highest percentage of viability was observed at 15 mg/kg and 20 mg/kg of AgNPs-loaded vaccine concentrations after injecting into the mice. The calculated potencies for alum-containing vaccine and AgNPs-loaded vaccine (dose 15 mg/kg) were 1.897 and 1.303, respectively. MTT assay demonstrated that alum at the concentration of 10 mg/mL was toxic, but AgNPs were not toxic. The in vivo toxicity also elucidated the safety of AgNPs and AgNPs-loaded vaccine in mice and dogs, respectively. Conclusion In the current study, for the first time, the adjuvanticity effect of green synthesized AgNPs on veterinary rabies vaccine potency with no in vivo toxicity was elucidated according to the European Pharmacopeia 8.0.

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Genome of the hepatitis C virus (HCV) contains a long open reading frame encoding a polypro tein that is cleaved into 10 proteins. Recently, a novel, so called "ARFP/F", or "core+1," protein, which is expressed through a ribosomal frame shift within the capsid coding sequence, has been described. Herein, to produce and characterize a recombinant form of this protein, the DNA sequence corresponding to the ARFP/F protein (amino acid 11-161) was amplified using a frame shifted forward primer exploiting the capsid sequence of the lb subtype as a template. The amplicon was cloned into the pET 24a vector and expressed in different Escherichia coli strains. The expressed protein (mostly as insoluble inclusion bodies) was purified under denaturing conditions on a nickel nitrilotriacetic acid (Ni NTA) affinity column in a sin gle step with a yield of 5 mg/L of culture media. After refolding steps, characterization of expressed ARFP/F was performed by SDS PAGE and Western blot assay using specific antibodies. Antigenic properties of the protein were verified by ELISA using HCV infected human sera and by its ability for a strong and specific interaction with sera of mice immunized with the peptide encoding a dominant ARFP/F B cell epitope. The antigenicity plot revealed 3 major antigenic domains in the first half of the ARFP/F sequence. Immunization of BALB/c mice with the ARFP/F protein elicited high titers of IgG indicating the relevance of produced protein for induction of a humoral response. In conclusion, possibility of ARFP/F expression with a high yield and immunogenic potency of this protein in a mouse model have been demonstrated.

The current experiment reveals the anticancer properties of silver nanoparticles (AgNPs) synthesized using aqueous leaf extract of Cichorium intybus, a significant medicinal plant. The characteristics of AgNPs were continuously studied by powder X-ray diffraction (XRD), Fourier-transform infrared spectroscopy (FTIR), zeta potential, transmission electron microscopy (TEM), scanning electron microscopy (SEM), and energy-dispersive spectroscopy (EDS) analysis. Current microscopic results show that produced AgNPs were spherical in shape with an average size of 17.17 nm. A strong peak between 2 and 4 keV showed the greatest ratio of the elemental silver signals, due to surface plasmon resonance (SPR). The AgNPs, fabricated by green method, had a negative zeta potential of - 9.76 mV, which indicates that the synthesized AgNPs is dispersed in the medium with high stability. The in vitro cytotoxicity effect of AgNPs showed promising anticancer activity against human breast cancer MCF-7 cells. Annexin V-FITC/propidium iodide assay, Hoechst 33258 staining, and upregulation of caspase 3 activity revealed significant apoptosis activities of AgNPs against MCF-7 cells. Moreover, the flow cytometric analyses of cell cycle distribution of MCF7 cells showed that AgNPs treatment has enhanced the sub-G1 peaks, which is an indicator of apoptosis pathway. Overall results in our study suggested that AgNPs fabricated by a biogreen approach could be useful in cancer therapy.

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