فهرست مطالب naser harzandi

Leptospirosis is an infectious zoonotic disease that can result in severe complications. It is widespread, especially in hot and humid climates such as the northern region of Iran. The immune responses to leptospirosis are multifaceted. Lipl41 is an outer membrane protein that is expressed during infection and is highly conserved among pathogenic species. This makes it a good candidate for diagnosis and induction of specific immune responses. The aim of the present study was to evaluate immune responses against recombinant Lipl41 in mice.

After immunizing of different groups of mice with recombinant Lipl41 (rLipl41), the levels of specific antibodies and cytokine profiles interferon-gamma/ interleukin-4 (IFN-γ/IL-4) were measured.

The results revealed that rLipl41 showed a significant increase in antibody levels compared with the control groups (P< 0.05). Although the level of IL-4 in the groups that received Lipl41 was similar to that in the other control groups, the IFN-γ levels showed a significant increase (P<0.05).

It has been concluded that recombinant Lipl41 protein could strongly stimulate specific immune responses and be considered a potential candidate for vaccine development and diagnostic research.

Antibiotic-resistant bacteria have increased in livestock and poultry over recent years. However, to date, there have been few studies that have investigated the antibiotic resistance bacteria among environmental and animal samples. Hence, the main purpose of this research was to isolate Enterobacteriaceae from the wastewater of poultry farms and investigate the pattern of antibiotic resistance in these bacteria. In this study, 100 wastewater samples were isolated from different poultry farms. These strains were identified using biochemical tests. The antibiotic resistance pattern was determined by the disk diffusion method (Kirby–Bauer). The isolated bacteria were investigated in terms of resistance patterns to cephalexin, tylosin, doxycycline, and erythromycin. A total of 170 Gram-negative bacteria from the Enterobacteriaceae family were isolated from poultry wastewater samples. The most isolated strain was Escherichia coli (35.33%), followed by Klebsiella (17.65%). According to this study, the most resistance was to cephalexin and doxycycline antibiotics. The results of this research indicate a high level of antibiotic resistance in bacterial isolates from wastewater. This is probably be due to the improper use of antibiotics in the poultry industry.

The most preferred method for the detection of foot-and-mouth disease (FMD) viral antigen and identification of viral serotype is the ELISA. Diagnostic tests with high sensitivity is necessary both to distinguish infected vaccinated animals and disease control programs for identifying the carrier animals. To detection of FMD virus, the current strategies are mainly based on capture antibody (sandwich) ELISA test. Applying laying pullets as animal bioreactor for production specific egg yolk antibody (IgY) has increased in recent years due to its high yield, affinity, low price, and quick production turnover. In the present study, we aimed to produce a concentrated and purifies IgY polyclonal antibody to design a capture antibody ELISA kit against the FMD virus (FMDV) serotype A. At first, laying hens were immunized by inactivated FMDV serotype virus and then, on days 14, 21, and 28 following vaccination, the eggs and sera were collected and then, the IgY polyclonal antibodies were extracted and purified from the chicken egg yolk using polyethylene glycol 6000–ethanol precipitation procedure. Extracts were filtered, purified by ion exchange chromatography and dialyzed. The purified IgY concentration estimated by Bradford assay, confirmed it presents by SDS-PAGE and Western blot and also its specific immune reaction by Ouchterlony double immunodiffusion and Dot blot tests. Moreover, for achieving optimum concentration of antigen/antibody (sera) in sandwich ELISA, checkerboard titration test was setup base on indirect ELISA results. Eventually, 119 previously confirmed samples (including 80 positive and 39 negative) by both Real time PCR (quantitative PCR, qPCR) and with a commercial ELISA kit, used for evaluation of sensitivity and accuracy of our developed Capture antibody ELISA kit. In this manner, the sensitivity and specificity of our designed kit was 100% and 98%, respectively. According to this, the present developed capture ELISA kit based on IgY has high sensitivity and specificity for FMD virus detection and it could be used in future as both commercial detecting and serotyping applications.

Despite widespread vaccination against foot-and-mouth disease, many outbreaks still occur in endemic areas. We attempted to determine the genetic and antigenic properties of the O/PanAsia-2/QOM-15 foot-and-mouth disease virus new vaccine strain. Thus, whole-genome sequencing was used to identify vulnerable pinpoint sites across the genome. The VP1 sequence (1D gene) of the O/PanAsia-2/QOM-15 viral genome was then compared to the VP1 sequences of two previously used vaccine strains, O/PanAsia (JQ321837) and O/PanAsia-2 (JN676146). The antigenic relationship of these three viruses was calculated by the two dimensional-virus neutralization test. At the nucleotide level, 47 single variants were identified, of which 19.00% were in the 5' untranslated region (UTR), 79.00% in the polyprotein region, and 2.00% in the 3' UTR region. Approximately half of the single nucleotide polymorphisms that have occurred in 1D gene resulted in amino acid (AA) substitutions in the VP1 structure. The single nucleotide polymorphisms also caused AA substitutions in other structural proteins, including VP2 and VP3, and some non-structural proteins (Lpro, 2C, and 3A). The O/PanAsia-2/QOM-15 shared higher sequence similarity with O/PanAsia-2 (91.00%) compared to O/PanAsia (87.30%). Evaluating r-value showed that the antigenic relationship of O/PanAsia-2/QOM-15 with O/PanAsia-2 (29.00%) was greater than that of the O/PanAsia (24.00%); however, all three viruses were immunologically distinct. After 10 years, the alteration of virus antigenicity and the lack of detectable adaptive pressure on VP1 sequence suggest that studying genetic dynamics beyond the VP1 region is necessary to evaluate FMDV pathogenicity and vaccine failure.

Hepatitis A virus (HAV) is a causative agent of acute hepatitis in humans, infecting more than one million individuals every year, including both symptomatic and asymptomatic infections. The currently available preventive vaccines for HAV are based on either wild-type or live-attenuated virus strains, which can contribute to the costliness of the vaccination process. Therefore, it may be worthwhile to explore the potential of subunit vaccines that utilize immunogenic viral products.

This study presents the results of a novel recombinant protein production study that employed the native structures of HAV-VP1 and HBs-Ag. The fusion protein underwent comprehensive characterization to evaluate its potential applications in diagnostics and immunization. The truncated recombinant protein, HAV-VP1 (position 99-259 aa) -HBs-Ag, was successfully expressed in the Escherichia coli BL21-DE3 system.

The recombinant protein, with a molecular weight of 46 kDa, was evaluated using SDS-PAGE gel electrophoresis and confirmed by western blotting. The fusion protein was successfully detected in serum samples positive for HBV or HAV using anti-HBs and anti-VP1 antibodies. Additionally, it elicited a potent humoral response in BALB/c mice.

The novel recombinant protein described in this study has the potential to serve as a bivalent vaccine against HAV and HBV infections. The next step involves evaluating the immunogenicity and safety profile of the protein.

Gardnerella vaginalis is one of the most important causes of prevalent genital infections that pose serious risks. This study aimed to determine the prevalence of Gardnerella vaginalis and antibiotic resistance pattern of isolates of patients referred to the gynecology clinic of Shahriar Noor Hospital by PCR and culture methods.

The study was conducted on 500 patients who had suffered from a vaginal infection. The demographic data of patients were studied. For diagnosis of Gardnerella vaginalis isolates, cultivation in anaerobic conditions, biochemical tests, PCR and Gardnerella vaginalis antibiotic susceptibility test to metronidazole and clindamycin were performed. Data analysis was performed utilizing SPSS statistical software version 19 and the Chi-square test.

Among the 500 patients, 173 were diagnosed with Gardnerella vaginitis. There was a significant relationship between age group, level of education, and contraceptive method with Gardnerella vaginosis incidence. Performing antibiotic susceptibility tests showed that the resistance of Gardnerella vaginalis isolated strains to metronidazole and clindamycin was 86.12% and 17.34%, respectively.

The high prevalence of Gardnerella vaginalis infections confirms the critical role of the bacterium in the occurrence of bacterial vaginosis. Therefore, it is necessary to check the prevalence of bacterial infections to recommend the correct medical treatment in different societies.

Colorectal Cancer (CRC) is the second most common cancer in men and women. Recently, investigations have revealed a much larger role for epigenetic and non-hereditary factors in CRC incidence than hereditary factors. Among all nonhereditary factors, gut microbiota alterations are the most prominent factor in the development of CRC. This work aimed to study the quantification of Enterococcus faecalis and Fusobacterium nucleatum in healthy colorectal tissues compared with polyp and cancer colorectal tissues of Iranian peoples.

In this case-control study, 21 biopsy samples of normal colon tissue, 21 polyp tissues, and 19 tumor tissues were investigated. To quantify the Enterococcus faecalis and Fusobacterium nucleatum in our samples, we employed the 16SrRNAspecific gene in Real-Time Quantitative PCR method.

The Quantitative Real-Time PCR results demonstrated a significant increase (P-value<0.05) in the population of both bacterial species, Enterococcus faecalis and Fusobacterium nucleatum in tumor and polyp tissues compared with normal samples. In addition, the Spearman index for these two species was 0.7634, which refers to a synergistic relationship between these species in the colon environment.

Collectively, by the progression of CRC, the abundance of Enterococcus faecalis and Fusobacterium nucleatum, will be increasing. In other words, the enrichment of these species can induce the development and progression of CRC and might be a sign of its occurrence.

Burkholderia mallei, is the etiologic agent of the glanders disease. Clinical and bacteriological diagnosis of glanders is difficult in the early stages of the disease. Some methods such as Complement fixation test (CFT) due to false positive results and troublesome for veterinary authorities and cause financial losses to animal owners, The other one is Malleination test, which requires appropriate equipment and efficient laboratory personnel. Therefore, in order to quickly and accurately diagnose the disease, especially in areas that cannot be kept animals, new methods should be used to identify the disease. The Western blot is a serological diagnostic test and has been recommended by the World Organization for Animal Health (OIE). In this study, a Western blot assay making use of a purified lipopoly-saccharide (LPS) containing antigen of Burkholderia mallei was designated.

in this study, a total of 75 sera were collected from different horse populations from several geographical areas of Iran. Specificity and sensitivity of the Complement fixation test, ELISA and a Western blot were compared for serodiagnosis of glanders. ELISA test was based on B. mallei antigens and Western blot made use of a purified LPS-containing B. mallei-antigen.

The Western blot and ELISA, were more specific than the Complement fixation test. ELISA based on B. mallei antigens had more sensitivity compared to Complement fixation test and Western blot. Finally, sensitivity and specificity were obtained for Complement fixation test (92.31% and 98.38%), Western blot (92.31%, 100%) and (100% and 100%) respectively.

Complement fixation test for glanders is still the prescribed serological method for commercial purposes, which is used to confirm the health of animals. However, in order to maintain the biosafety of valuable and expensive horse colonies, it is important to implement a more accurate and intelligent disease control and eradication program. This enhances the laboratory diagnostic capabilities to better understand the cause of the disease and to use and optimize the diagnostic methods of glanders in the country. Therefore, efforts to further improve and optimize ELISA and Western blotting should be continue.

European honey bees (Apis mellifera) are the most important pollinator insects that play vital role in maintenance of all most all life forms on earth. However, over the last decade major concerns have raised due to decline in the population of these insect species. A variety of factors have been responsible for these concerns of which the most important is honey bee bacterial diseases like Nosemosis (Nosema), American and European foulbrood diseases. While, overuse of antibiotics utilized for the treatment and control of these diseases has resulted in the emergence of antibiotic resistant strains of these pathogens. Probiotics have been considered a suitable substitute of antibiotics in human and animals. In last several years, lactobacillus species isolated from honeybees have been considered of significance in enhancing the life span of honeybees by reducing the incidence of bacterial and viral infections in these tiny insects. Among the isolated microbes in the gut of honeybees, Lactic Acid Bacteria (LAB) are of utmost importance showing direct impacts on the health of their host by modulating the gut microbial flora and are termed as Probiotic bacteria.The objective of this research was to isolate and identify LAB from different parts of the intestinal tract of honeybees Apis mellifera and to characterize their probiotic properties.

Twenty-four honeybees collected from the hives located in the city of karaj were analyzed for the presence of LAB species. The stomach contents of honeybees were inoculated into MRS broth, incubated at 37ºCfor 48 hrs. The obtained colonies were purified and identified to species levels phenotypically and geno-typically. Hemolytic activity and sugar fermentation reactions of the isolates were recorded and later subjected 16SrRNA sequencing using a pair of universal primers. The identified isolates were evaluated for their viability in acidic conditions at pH 2.5, 3.0, 4.0 and 6.5 at different time intervals. Bile resistance of the isolates was tested by culturing the isolates in the presence of different concentrations of the said salts (0.5, 0.7 and 1%). Survival of LAB isolates in simulated gastric and intestinal conditions containing different enzymes and bile salts, antibacterial spectrum against a number of gram positive and gram-negative pathogens by agar well diffusion assay, and their in vitro colonization ability (aggregation, co-aggregation and hydrophobicity percentages) were evaluated. The results were analyzed statistically.

Twenty nine gram positive, catalase negative and non-hemolytic colonies were isolated from 24 honeybee samples. Among these, only 7 colonies showed enhanced antibacterial activity and were selected for further studies. Based on phenotypic characteristics, sugar fermentation reactions and 16S rRNA sequencing the isolates were identified as Lactobacillus acidophilus (1), Lacticaseiobacillus casei, Lactiplantibacillus plantarum (2), Lactobacillus apis (1), Enterococcus faecium (1) and Pediococcus acidilactici (1). During probiotic characterizations, the identified isolates were shown to retain their viability in acidic conditions and resisted pH 2.5, 3 and 4 for more than 4 hrs. However, slight decrease in viability at pH 2.5 and 3.0 was observed, compared to pH 4.0 and above. All isolates appeared bile resistant and tolerated all used concentrations of bile salts during 8 hrs of incubation. Survival rate of the isolates in simulated intestinal conditions was significantly (p˂0.05) greater compared to simulated gastric conditions indicating greater stability of the isolates to alkaline conditions rather than to acidic conditions. L.acidophilus and E.faecium showed least resistance in gastric conditions and their growth rate was decreased more than 50% under said conditions. In contrast, the growth rate of these two isolates was highest in simulated intestinal conditions as they resisted these conditions for more than 24 hours.  The isolates demonstrated antibacterial affect against a number of tested pathogens including Listeria monocytogenes, Escherichia coli, Enterococcus faecalis and Streprtococcus mutans. The auto-aggregation, and cell surface hydrophobicity percentages of L.casei appeared highest compared to other tested Lactic Acid bacteria in study (p˂0.05), while, L.apis showed the highest co-aggregation with S.typhi strain. P.acidilactici possessed the least auto-aggregation (46%), co-aggregation (10%) and hydrophobicity (43%) percentage. Auto-aggregation ability appeared directly related to hydrophobicity percentages and isolates showing high aggregation ability also showed high hydrophobicity %.

In conclusion, honeybee gut appeared a reservoir of LAB with probiotic potential. It is suggested that further studies should be conducted in order to determine the health benefits of these LABs in honeybees, with especial emphasis on their ability to prevent honeybee diseases.

Leptospirosis is a zoonotic disease caused by pathogenic spirochete called Leptospira that has worldwide distribution. Unfortunately, due to the variety of clinical symptoms, its diagnosis has many limitations. LipL41 is among the most abundant outer membrane proteins in Leptospira and is exclusively found in pathogenic species. A small gene called lep is located very close to lipl41 gene on the bacterial chromosome, which facilitates its expression. In this study, the expression and purification of LipL41-Lep fusion protein among prevalent pathogenic isolates in Iran was performed in a prokaryotic system for the first time.

All collected Lipl41 and Lep protein sequences were analyzed from the NCBI database. Complete codon sequences of the pattern of the Iranian serovars were designed and synthesized then sub-cloned into a pET32a+ and transformed into Escherichia coli BL21(DE3) to expression by using IPTG inducer. Thefusion recombinant protein was purified by denaturation and confirmed by western blot.

The results of PCR analysis showed a clear band of ~ 2000 bp in Agarose gel. Optimal expression of fusion recombinant protein (60kDa) was achieved post-induction at 4 h at 37 °C in the presence of 0.1 mM IPTG. Then it was purified in the insoluble form.

The results demonstrated that adequate amounts of this fusion recombinant protein were expressed andpurified to be used as a fusion antigen in serological testing, such as ELISA, or for the development of subunit vaccines against Leptospirosis in the future.

The use of chemical anti-cancer drugs frequently create serious side effects. However, probiotics are natural and treat different kinds of cancer without undesired effects. In this study, a nano delivery system was planned to transport the Lactobacillus rhamnosus GG (L. GG) cytoplasmic fraction (Cf) to cancerous tissue in the mouse model.

Magnetic iron nanoparticles (MINPs) were synthesized and loaded with L. GG-Cf (0, 0.312, 0.625, 1.25, 2.5 mg/ml) and administrated for three weeks to treat experimentally induced murine breast cancer in a constant magnetic field. At the end of the trial, the treating efficacy of this complex molecule was evaluated via western blotting and qPCR.

Results showed MINPS can deliver and accumulate the L. GG-Cf in cancer tissue, also the size and volume of the tumors were reduced. Additionally, in cancer tissues of treated mice with 2.5 mg/ml of Cf-MINPs significant induced apoptosis was seen compared to untreated (control), and our data proved that this induction may be due to the caspase-3 pathway.

In conclusion, L. GG-Cf could treat the murine breast cancer and MINPs are a suitable candidate for drug delivery because of their safety, uniformity, and magnetic properties.

Use of chemical anti-cancer drugs frequently creates serious side effects. However, probiotics are natural and treat different kinds of cancer without undesired effects. In this study, a nano delivery system was planned to transport the Lactobacillus rhamnosus GG (L. GG) cytoplasmic fraction (Cf) to cancerous tissue in a mouse model. Magnetic iron nanoparticles (MINPs) were synthesized and loaded with L. GG-Cf(0, 0.312, 0.625, 1.25, and 2.5 mg/ml) and were administrated for three weeks to treat experimentally induced murine breast cancer in a constant magnetic field. At the end of the trial, the treating efficacy of this complex molecule was evaluated via western blotting, immunohistochemistry, and qPCR. Results showed that MINPS can deliver and accumulate L. GG-Cf in cancer tissue, and reduce the size and volume of the tumors. Additionally, in cancer tissues of treated mice with 2.5 mg/ml of Cf-MINPs, significantly induced apoptosis was seen compared with untreated mice (control), and our data proved that this induction may be due to the caspase-3 pathway. L. GG-Cf could treat murine breast cancer, and MINPs are a suitable candidate for drug delivery because of their safety, uniformity, and magnetic properties.

Brucella spp. are intracellular pathogens, therefore cell-mediated immunity is the main response to inhibit survival and growth of the bacteria in vertebrate host. Many eukaryotic plasmid vectors are being used in setting up DNA vaccines which may show different efficiencies in same conditions. This is important in designing the vaccines and immunization strategies. We looked into the probable differences of immune responses induced by different eukaryotic DNA plasmid vectors (pcDNA3.1 and pVAX1) harboring the same Omp31 gene of B. melitensis. Female BALB/c mice were immunized with pcDNA-omp31 and pVAX-omp31 and further boosted with recombinant Omp31. Subclasses of specific serum IgG against the rOmp31 were measured by ELISA. Cytokines responses to rOmp31 in Splenocyte cultures of the immunized mice were evaluated by measuring the production of IL4, IL-10, IL-12 and IFN-γ. Protective responses of the immunized mice were evaluated by intraperitoneal challenge with pathogenic Brucella melitensis 16M and Brucella ovis PA76250. Both DNA vaccine candidates conferred potent Th1-type responses with higher levels of cytokines and immunoglobulins observed in mice immunized with pVAX-omp31. Although pcDNA-omp31 and pVAX-omp31 both elicited protective immunity, mice immunized with the latter showed a higher protection against both B. melitensis and B. ovis PA76250. The results of this study highlight the significant differences between efficiency of diverse plasmid backbones in DNA vaccines which code for an identical antigen. Comparing various plasmid vectors should be considered as an essential part of the studies aiming construction of DNA vaccines for intracellular pathogens.

Saffold virus as a new member of cardiovirus genus in picornaviridae family has been suggested to be related to diarrheic cases and human airway diseases. However, relationship between Saffold virus and human diseases is unclear. In order to establish an investigation for the occurrence of Saffold virus among pediatric patients involved to acute gastroenteritis, we implemented a RT-PCR assay for detection and quantification of Saffold virus in stool specimens.

In this study, a total of 160 stool samples from September 2018 to May 2019 were collected from presenting pediatric patients with acute gastroenteritis in a Karaj hospital, Iran. After viral RNA extraction, the RT-PCR was performed to amplify the 5’UTR region of Saffold virus genome.

Out of the 160 samples tested, the Saffold virus genomic RNA was detected in 26/160 (16.2%) of stool samples. The high Saffold virus detection rate was related to February (6/26 or 23%). The co-infection of Saffold virus with Aichivirus and Salivirus as other new emerging viruses was also assessed, among which high double or triple mixed-infections were determined.

This is the first documentation of Saffold virus detection in stool samples that demonstrates Saffold virus has been circulating among Iranian pediatric patients. Our results indicated that Saffold virus in association with Aichivirus and Salivirus may be possibly considered as causative agent of acute gastroenteritis.

The Enterococcus faecalis (E. faecalis) is the one of the pathogenic bacteria that become famous and considerable in the recent years. Here we tried to do typing the E. faecalis isolates to provide advantageous information that can help us to understand epidemiological communication between the E. faecalis isolates.

One hundred  E. faecalis were isolated from urine samples of Imam Khomeini Hospital, Ilam, Iran. Afterwards, all isolates were confirmed by the phenotyping method and then for more certainty, every isolates were authenticated by PCR analysis of 16sRNA gene. Eventually, all isolates were considered as E. faecalis. For Multilocus Sequence Typing (MLST), 7 housekeeping genes were used to gain MLST scheme for epidemiological study. In addition, to determine various type of E. faecalis pubmlst database was selected and the MLST analysis was done based on recommended instruction by the pubMLST.org.

The disk diffusion results demonstrated that fifty-four out of one hundred isolates were resistant, four isolates were semi sensitive and forty-two isolates were sensitive to vancomycin. So, 90 isolates were MLST. Using seven structural genes and using pubMLST.org database, different types of E. faecalis were determined. The MLST results demonstrated that 26 different group and Sequence Types (ST) obtained. Our findings demonstrated that the isolates were from different types.

 Accoeding to our results, we couldnchr('39')t find any epidemic correlation between the isolates. Given that most of these isolates had resistance to vancomycin, they had low clonal correlation with each other and only had few similar STs pattern.

Echium amoenum (E. amoenum), as one of the most popular plants in Iran, is traditionally used to treat different types of disorders.

This experimental study aimed to evaluate the modulatory effects of E. amoenum on permethrin (PMN)-induced oxidative stress in rats and to determine the cytoprotective effect of E. amoenum on PMN in SK-Hep-1 cells.

Twenty-four male Wistar rats were randomly divided into four equal groups, including the control (normal saline), orally treated PMN (125 mg/kg of PMN), E. amoenum (100 mg/kg), and E. amoenum + PMN groups for 28 days. The levels of alanine aminotransferase (ALT), alkaline phosphatase (ALP), aspartate aminotransferase (AST), lipid peroxidation (LPO), catalase (CAT), and glutathione peroxidase (GPx), as well as the expression of catalase (CAT) and glutathione peroxidase (GPx), were measured in the liver of all rats. Also, the cytoprotective effect of E. amoenum against PMN was evaluated in the treated SK-Hep-1 cells.

The results indicated that LPO increased significantly in the PMN-treated group, as evidenced by the high concentration of malondialdehyde (MDA) in the liver. Alterations of the antioxidant system were also confirmed by the significant decline in CAT and GPx activities (2.9 ± 0.14 and 0.5 ± 0.03, respectively; P < 0.05) and the significant downregulation of CAT (0.4 ± 0.02 folds) and GPx (0.3 ± 0.01 folds) mRNA expression in the liver (P < 0.05). PMN also stimulated significant changes in hepatic biomarkers and induced pathological changes in the liver. On the other hand, administration of E. amoenum significantly reduced abnormalities in biochemical markers, LPO, antioxidant enzymes, gene expression, and pathological complications induced by PMN (P < 0.05). E. amoenum also exhibited cytoprotective effects against cytotoxicity induced by PMN in SK-Hep-1 cells.

The present results demonstrated that E. amoenum has significant antioxidant, gene-regulating, and cytoprotective effects.

Nanosilver and nanocurcumin are popular nonmaterial with increasing public attention, but their synergistic therapeutic potentials in burns have never been considered. The present study aimed to provide a novel formulation from both nanomaterials (Ag – Curcumin NPs) and conduct their preclinical evaluation for burn healing. After evaluation of particle size, loading efficiency, release profile and morphology of manufactured nanoparticles by TEM and DLS techniques , a 14 days skin irritation and corrosion test on Albino rabbits was performed based on OECD 404 guideline . The Ames Mutagenicity test was performed on 4 strains of Salmonella Typhymurium after MIC and MBC adjustments indifferent doses. For clinical efficacy, skin burn model was designed and applied in rats by providing limited standard 3rd degree burns at the back of each rabbit. Manufacture lyophilized spherical nanoparticles in the range of 20- 38 nm, with low zeta potential didn't show any significant size enlargement. The nanogel was also considered as a nonirritant and non-corrosive formulation after short term and long term dermal applications (>72 hrs.) .No mutagenic effects were also identified in all strains in the test samples of Ag - Curcumin NPs (Mutation Index=0.08-0.27). This study clearly showed the safety of Ag – Curcumin NPs nanogel in low concentrations with small dimension (16-32μg/ml). Due to the safety of proposed formulation and increased rate of wound healing by Ag-Curcumin nanogel in comparison to both control groups, this combinational formulation could be considered as a safe and effective candidate for further clinical applications.

Production of viral vaccines and also drugs products which are derive from cells produced in various biological materials, like cell substrates eg: fertile eggs, primary cells, cell lines, raw materials and additives used in culture media which have animal origin and seed virus which is used for culture have a complicated process. The above processes are naturally vulnerable for contamination to adventitious agents. The Nanobacteria like other adventitious agents can cause contamination through primary materials with animal origin for such a product. The aim of this study was to design a program for PCR which is specific for detection of probable presence of Nanobacteria in cells, especially in cell lines and diploids which is used in vaccine production in Razi institute. For this purpose the cells which are used in following departments, research, production and Quality control for biological products were collected and DNA was extracted for PCR expression on 16S rRNA area from Nanobacteria. Bartonella henselae was used as a positive control. As it was expected the results of PCR products after electrophoresis were positive for above organism and at the area of 558 bp replicated which it confirms the correctness of our experiment. The cells from all departments were negative for Nanobacteria, however for not having enough information for spread and risk evaluation of Nanobacteria in cattle population there are a great need for further investigation.

Silver nanoparticles (AgNPs) are one of the most important nanoparticles which have various biomedical applications. For example as antifungal, antibacterial, anti-cancer and anti-inflammatory agents. Skin infection caused by Trichophyton rubrum and some opportunistic fungi such as Candida albicans and Aspergillus fumigatus are sometimes difficult to be treat. Although silver nanoparticle has long been used as effective inorganic antifungal agent; the antifungal activity of nano-Ag in different size has not been investigated yet. Anti-cancer and antifungal effects of spherical silver nanoparticles (nano-Ag) were investigated in this study and we decided to determination toxicity of Nano-Ag in different diameters (10, 20 and 40 nm) on L929 mouse fibroblasts. TEM microscope has been used to evaluate nanoparticles size and morphology. Nano-Ag’s toxicity were evaluated by MTT assay [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide]. This study showed that the toxicity of Nano-Ag in low concentration (16-32µg/ml) are sustainable and Silver nanoparticle effects are size dependent.

     Multidrug resistance is a serious problem in the treatment of urinary tract infections. Integrons are ancient structures that contain determinants of a site-specific recombination system to capture genes encoding antimicrobial resistance. The aim of this study was to determine the prevalence of multidrug resistant Klebsiella isolates in clinical specimens. In addition, the existence of integrons in resistant isolates was assessed by amplification of integrase genes.

  Susceptibility of 100 Klebsiella isolates was determined to 19 antibiotics by the Kirby-Bauer disc diffusion method and integron classes were detected. Then the multi-drug resistance association with the existence of the integrase gene was calculated by chi-squre and fisher exact tests.

   According to antibiogram results, 54 isolates were multidrug resistant. By amplification of intergrase gene it was revealed that 27% of the isolates harbor integrons. Also by PCR-RFLP it was revealed that all of them were class 1. The existence of integrons was confirmed in 25 of our multidrug resistant isolates, indicating the frequency rate is high and integrons may be partly responsible for multidrug resistant.

  Multidrug resistance suggests that strategy for treatment of patients with Klebsiella infections needs to be revised. The possibility of transmission of resistance genes by integrons would be decreased by treatment of patients with the appropriate antibiotics.

Non-typhoidal salmonellae and serovars of Escherichia coli, Listeria, Shigella, and Campylobacter are among the most important food-borne bacterial agents. The aim of the present study was molecular detection of quinolone resistance in Salmonella spp. isolated from poultry products in Karaj, Iran, using polymerase chain reaction (PCR). Ninety samples of poultry products were collected from different brands and markets during September-December 2017. All samples were enriched in nutrient broth and Selenite F broth, and salmonellae were isolated by xylose lysine deoxycholate agar. The presence of specific genes of qnrA, qnrB, and qnrS was investigated employing PCR technique and subsequently, specific primers. None of the 30 egg yolk samples had bacterial growth in the culture medium. In total, 29 (48.33%) out of 60 raw chicken meat samples were determined to be contaminated with Salmonella using culture-based methods (i.e., 7 (35%) out of 20 drumstick, 15 (62.5%) out of 24 breast, and 7 (43.75%) out of 16 liver samples). In addition, the frequency of qnrA, qnrB, and qnrS genes in the samples was 10.34%, 68.96%, and 86.20%, respectively. The results of this study showed a high frequency of Salmonella contamination and qnr genes in the contaminated samples.

Bacillus thuringiensis var kurstaki is one of the best known biological insecticide which is used extensively in the world. This biopesticide has been broadly used against important insect pests and vectors of animals and humans. However, the controversial data was published to date considering toxicological studies on non-target species are urgent to determine probable adverse effects and risk assessment of this biopesticide. In this research, histopathological changes, hematological and some biochemical factors were evaluated following the single dose oral administration of B. thuringiensis in rats. Twelve Wistar rats were randomly divided into 2 groups including experimental and control groups. Animals were treated orally by gavage to single dose of sub-lethal dose (5000 mg/kg) of B. thuringiensis suspension. Finally, hematological factors (WBC, RBC, Hb, HCT, MCV, MCH, MCHC and PLT), some biochemical parameters (ALT, AST, ALP, BUN and Cr) and histopathological observations were evaluated. The results demonstrated that oral administration of high dose of B. thuringiensis biopesticide induced some pathological complications including congestion and inflammation in vital organs of rats such as liver, heart, lung and kidney. It also caused hematological abnormalities and biochemical alterations in the experimental animals group. According to the findings, it can be concluded that high dose of B. thuringiensis biopesticide can induce toxicity in rats. Therefore, further investigations including subacute and chronic are recommended.

Human group A rotaviruses are the most common cause of severe diarrhea among children This cross-sectional study was carried out on 76 samples collected from both influent and effluent water of 4 sewage treatment systems, hospital sewage, Karaj and Baraghan rivers in Alborz Province. All samples were concentrated using pellet method. Afterwards, human group A rotaviruses were detected using ELISA method.
In total, rotaviruses were identified in 6 samples (7.89%) using ELISA method. A significant relationship was found between rotavirus detection and sampling site (P=0.039). The frequency of rotavirus detection in summer, autumn and winter was 1 sample (16.66%), 3 samples (50%) and 2 samples (33.33%), respectively.
This study showed the contamination of environmental water by human group A rotaviruses. Therefore, it is necessary to constantly monitor sewage treatment systems and river water for detecting rotaviruses.

Shigella bacteria can infect human body by taking contaminated food and water and are transmitted from person to person. Human body is the only natural host for these bacteria. The aim of this study was to detect Shigella contamination in pre-packed samples of salads at restaurants in western regions of Tehran city using PCR method.
To conduct this research, 90 samples were purchased from the restaurants during the period of June to November 2016. The samples were cut into very small pieces, homogenized and a 25g portion of these samples was added to 225 ml of Shigella broth media containing novobiocin and incubated for 24 hours. Then DNA of cultured samples was extracted using DNPTM kit (CinnaGen, Iran) and PCR method was optimized for amplification of 613 bp segment of ipaH gene and performed on extracted DNA of all samples (before and after enrichment in Shigella broth).
Shigella contamination was detected in 7(7.8%) and 20(22.2%) of the tested samples before and after the enrichment, respectively.
The results showed the contamination with Shigellabacteria in remarkable percentage of the samples and revealed the necessity of more attention and supervision in the processes of production and distribution of pre-packed salads. Besides, the findings points to the importance of enrichment in increasing the sensitivity of the PCR method to detect the bacteria in food samples.