[Identification of enterotoxigenic Escherichia coli strains in children under 2 years of age by real-time PCR in Shiraz]

Enterotoxigenic strains of E. coli [ETEC] form the second largest group of diarrheagenic E. coli and are responsible for more than 25% of infantile diseases. The purpose of this study is to assess the prevalence of ETEC strains and their virulence genes in children less than 2 years of age in Shiraz. This was a cross-sectional study. 285 stool samples were taken from children under 2 years of age with diarrhea in Shiraz. First, E. coli strains were isolated using standard biochemical tests. Then, prevalence of ETEC strains and presence of st and lt genes were evaluated using real-time PCR. Antibiotic susceptibility of the isolates was evaluated by using disc diffusion method. 49 [17%] diarrheagenic E. coli strains were isolated, 7 cases [14/3%] belonged to ETEC strain. Among isolated ETEC strains, 4 [57.14%] had lt, 1 [14.29%] had stIa and 2 strains [28.57%] had both lt and stIa genes. All ETEC strains were susceptible to chloramphenicol, amikacin and nitrofurantoin, but showed resistance to penicillin and macrolides. The results showed high prevalence of ETEC strains in our study area. We recommend hospital-wide surveillance by using molecular techniques to estimate the prevalence of this pathotype in other regions of our country

Construction of knowledge, attitude and practice questionnaire for assessing plagiarism

This study was conducted to develop a questionnaire in order to evaluate knowledge, attitude and practice [KAP] of the faculty members and students toward plagiarism. A KAP study was conducted from June to October 2011 enrolling 390 volunteers anonymously [response rate 96%]. The questionnaire included the following four parts: [a] general characteristics like gender, academic degree and education level; [b] nine questions regarding knowledge [Min=0, Max=9]; [c] nine questions regarding attitude [Min=9, Max=27]; and [d] eight questions regarding practice [Min=0, Max=8]. A pilot study was conducted to assess reliability of the questions regarding knowledge and attitude. Cronbach's alpha coefficient for the knowledge and attitude questions was 0.70 and 0.74 respectively. The overall prevalence of at least once plagiarism commission was 38% [SD=0.035]. The overall mean score of knowledge, attitude and practice was 5.94 [SD=1.66], 24.12 [SD=2.99], and 0.66 [SD=1.15] respectively. Knowledge of plagiarism was significantly higher among higher academic degrees and females. Their negative attitude toward plagiarism was stronger too. No statistically significant difference regarding plagiarism commission was observed among different academic degrees in both sexes. According to linear regression analysis, plagiarism commission decreased 13% per one unit increase in score of knowledge [P=0.005] and 16% per one unit increase in score of attitude [P<0.001]. This knowledge, attitude, and practice [KAP] questionnaire was developed as a standard tool in order to assess perception of subjects toward plagiarism and to estimate the prevalence and the type of plagiarism commission

[Isolation of Escherichia coli O[157]: H[7] in sheep meats using cultural and PCR method]

Background and aim: Escherichia coli O[157]:H[7] is now recognized as an important cause of diarrhea, hemorrhagia colitis and hemolytic-uremic syndrome worldwide. Meat contaminated with animal feces is probably the major source of the E. coli O[157]:H[7]. In this study, we investigated the prevalence of E. coli O[157]:H[7] in meat samples of sheep in Isfahan from August 2008 to January 2010

Methods: A total of 148 sheep carcasses in Isfahan slaughterhouse were assessed for E. coli O[157]:H[7] using the standard cultural and polymerase chain reaction [PCR] methods. Bacteriological examinations were performed by using Triptone Soya Broth containing Novobiocin [TSB-n] as an enrichment media and then sorbitol MacConkey agar plates supplemented with Cefixime and Tellurite [CT-SMAC] a selective plating media. Suspected colonies of E. coli O[157]:H[7], identified by bacteriological methods, were tested by PCR

Results: Using cultural method, 43 [29.1%] and 10 [6.8%] samples were positive for E. coli and E. coli O[157]:H[7], respectively. Only 5 sorobitol negative E. coli strains were identified as E. coli O[157]:H[7], using polymerase chain reaction. The seasonal prevalence of E. coli O[157]:H[7] in samples were 0-9.7% and it was at its highest level in Spring and Summer

Conclusion: These results indicate that sheep can be a reservoir for E. coli O[157]:H[7] and sheep meat may serve as a vehicle for the pathogen transmission to human

[Construction of a novel recombinant vector carrying herpes simplex virus thimidine kinase and yeast cytosine deaminase suicide genes to manipulate leishmania genetically]

Leishmaniasis is a group of diseases with various clinical pictures, which is caused by Leishmania spp. This parasite causes disease in human and about one hundred animal species. The disease is widely distributed in Iran and also across the world. One of the best approaches in the development of vaccine against Leishmania is genetically modification of the parasite. Therefore, the aim of this study was to design a novel genetic construct in order to insert Herpes Simplex Virus thimidine kinase [HSV-tk] and Yeast cytosine deaminase [Yeast-cd] suicide genes into the Leishmania genome. In this work, at the first step, HSV-tk and Yeast-cd fragments along with a gene of Leishmani, alpha-tubuline were cloned into pBluescript vector and the arrangement of tk-aplha tub-cd was created. Subsequently, these fragments were cut off from the plasmid and sub cloned into the plasmid pF4X1.4.4sat. The final construct was confirmed by digestion with relevant restriction enzymes. The fragments tk-alpha tub-cd was cloned successfully in the plasmid pBluescript and its authenticity was confirmed. Subsequently, this genetic collection was inserted into the plasmid pF4X1.4sat and finally, the orientation of it was checked. The construct designed in this study was able to insert two cellular suicide genes HSV-tk and Yeast-cd into the Leishmanai genome. Thus, this is an approach in achievement of vaccine against Leishmanai in the coming up researches

[Evaluation of immunogenicity of divalent DNA vaccine encoding brucella melitensis omp31 and P39 genes in balb/c mice]

Brucella is a facultative intracellular pathogen and one of the etiologic agents of brucellosis that can infect humans and domestic animals. Attenuated strains such as B. melitensis Rve1 and B. abortus S19 and Rb51 are being used to control brucellosis in domestic animals. However, no safe and effective vaccine is available for human use. This study was designed to evaluate the immunogenicity and the protective efficacy of a divalent fusion DNA vaccine encoding both the B. melitensis Omp31 protein and P39 protein, designated Pcdna3 recombinant vector. This experimental study was performed in Biotechnology Research Center of Islamic Azad University, Shahrekord branch in summer, 1386. Construction of pCDNA3 recombinant vector containing Omp31 and P39 genes of B. melitensis was completed. Then, 12 Balb/c mice were immunized intramuscularly with 100 mg per 50 micro liters of this DNA vaccine. Control mice, 12 Balb/c mice, were simultaneously injected with PBS. During the 1[st], 7[th], 15[th] and 30[th] days the mice received the injections. Afterwards, the ELISA cytokine assay was performed and data were analyzed by SPSS software. Intramuscular injection of the divalent DNA vaccine elicited cellular immune responses in Balb/c mice. The ELISA cytokine assay with serum of vaccinated mice showed high level of IFN-y and low changes of IL-4 in compare with control mice. Use of divalent genetic vaccine based on the Omp31 and P39 genes can elicit a strong cellular immune response against Brucellosis