Golestan University of Medical Sciences Medical Laboratory Journal 2538-4449 10 6 2016 11 1 Evaluation of Prevalence of S. pneumoniae pharyngeal carriers under 5 years of age by lytA gene detection 1 6 FA Fatane Moein Jahromi Department of Microbiology Jahrom Branch Mohammad Kargar Department of Microbiology Abbas Doosti Department of Marine Microbiology, Persian Gulf Marine Biotechnology Medicine Research Center, Bushehr University of Medical Sciences, Bushehr, Iran Zahra Mohammadalipour Department of Microbiology, Jahrom Branch ABSTRACT          Background and Objective: Streptococcus pneumoniae is one of the leading causes of death among children worldwide. Nasopharyngeal colonization in children can spread pneumococcal infections in the community. This study aimed to evaluate the prevalence of S. pneumoniae strains isolated from healthy pharyngeal carriers less than 5 years of age.           Methods: This cross-sectional descriptive study was performed on 150 children under 5 years old in the city of Shiraz. After nasopharyngeal swab sampling, the samples were cultured on blood agar containing 5% sheep blood. The cultures were incubated at 37 °C for 24 h. Primary identification was carried out using optochin sensitivity testing, bile solubility testing and gram staining. Molecular identification of S. pneumoniae strains was done using lytA gene-specific primers.         Results: Of the 150 samples collected from healthy children, 24.67% were pharyngeal carriers of S. pneumoniae. The highest frequency of pneumococcal strains was related to male carriers (n= 22, 59.46%) and the children aged 1-2 years (n=11, 29.73%). The results showed no significant association between the prevalence of pharyngeal carriage and gender or age.           Conclusion: Given the increasing rate of pharyngeal carriage of S. pneumoniae in children as a risk factor for respiratory tract infections, there is a need for further monitoring of the circulating serotypes and investigation of antibiotic-resistance mechanisms.          Keywords: Streptococcus Pneumoniae, Pharyngeal Carriers, lytA. ABSTRACT          Background and Objective: Streptococcus pneumoniae is one of the leading causes of death among children worldwide. Nasopharyngeal colonization in children can spread pneumococcal infections in the community. This study aimed to evaluate the prevalence of S. pneumoniae strains isolated from healthy pharyngeal carriers less than 5 years of age.           Methods: This cross-sectional descriptive study was performed on 150 children under 5 years old in the city of Shiraz. After nasopharyngeal swab sampling, the samples were cultured on blood agar containing 5% sheep blood. The cultures were incubated at 37 °C for 24 h. Primary identification was carried out using optochin sensitivity testing, bile solubility testing and gram staining. Molecular identification of S. pneumoniae strains was done using lytA gene-specific primers.         Results: Of the 150 samples collected from healthy children, 24.67% were pharyngeal carriers of S. pneumoniae. The highest frequency of pneumococcal strains was related to male carriers (n= 22, 59.46%) and the children aged 1-2 years (n=11, 29.73%). The results showed no significant association between the prevalence of pharyngeal carriage and gender or age.           Conclusion: Given the increasing rate of pharyngeal carriage of S. pneumoniae in children as a risk factor for respiratory tract infections, there is a need for further monitoring of the circulating serotypes and investigation of antibiotic-resistance mechanisms.          Keywords: Streptococcus Pneumoniae, Pharyngeal Carriers, lytA. ABSTRACT          Background and Objective: Streptococcus pneumoniae is one of the leading causes of death among children worldwide. Nasopharyngeal colonization in children can spread pneumococcal infections in the community. This study aimed to evaluate the prevalence of S. pneumoniae strains isolated from healthy pharyngeal carriers less than 5 years of age.           Methods: This cross-sectional descriptive study was performed on 150 children under 5 years old in the city of Shiraz. After nasopharyngeal swab sampling, the samples were cultured on blood agar containing 5% sheep blood. The cultures were incubated at 37 °C for 24 h. Primary identification was carried out using optochin sensitivity testing, bile solubility testing and gram staining. Molecular identification of S. pneumoniae strains was done using lytA gene-specific primers.         Results: Of the 150 samples collected from healthy children, 24.67% were pharyngeal carriers of S. pneumoniae. The highest frequency of pneumococcal strains was related to male carriers (n= 22, 59.46%) and the children aged 1-2 years (n=11, 29.73%). The results showed no significant association between the prevalence of pharyngeal carriage and gender or age.           Conclusion: Given the increasing rate of pharyngeal carriage of S. pneumoniae in children as a risk factor for respiratory tract infections, there is a need for further monitoring of the circulating serotypes and investigation of antibiotic-resistance mechanisms.          Keywords: Streptococcus Pneumoniae, Pharyngeal Carriers, lytA.

Golestan University of Medical Sciences Medical Laboratory Journal 2538-4449 10 6 2016 11 1 Inhibitory Potential of Metabolites from Lactobacillus casei against Phenotypic Characteristics of Salmonella typhimurium LT2 7 13 FA Hossein Naghili PhD of Food Hygiene Hossein Tajik Department of Food Hygiene and Quality Control, Faculty of Veterinary Medicine Mojtaba Raeisi Department of Health, Faculty of Health, Hadi Ghasem Mahdi Department of Food Hygiene and Quality Control, Faculty of Veterinary Medicine Mehran Moradi Department of Food Hygiene and Quality Control, Faculty of Veterinary Medicine Majid Amin zare Department of Food Hygiene and Quality Control, Faculty of Veterinary Medicine Touraj Mehdizadeh Department of Food Hygiene and Quality Control, Faculty of Veterinary Medicine Hasan Hasanzadazar Department of Food Hygiene and Quality Control Fardin Hariri Doctor of Veterinary Medicine and Master of public health ABSTRACT           Background and Objective: Several studies have shown the antimicrobial activity of lactobacilli against Salmonella enterica (serotype typhimurium). The aim of this study was to evaluate the inhibitory potential of metabolites produced by probiotic culture of Lactobacillus casei against S. typhimurium and its impact on S. typhimurium motility and biofilm formation.          Methods: In order to evaluate the impact of the metabolites, L. casei cell-free culture supernatant (CFCS) was collected by centrifugation of L. casei secondary cultures. Effectiveness of the CFCS against Salmonella was evaluated by the well-diffusion method. In addition, in vitro effect of this treatment on motility in Swarm agar and biofilm formation by the bacteria was investigated.          Results: Inhibition zone diameters of S. typhimurium were 0.83 and 12.1 mm at concentrations of 50 and 100 μl of Lactobacillus CFCS against the log4 of S. typhimurium, respectively. Moreover, CFCS treatment inhibited the motility and biofilm formation by Salmonella. Concentrations of 5% and 10% were determined as the minimum inhibitory concentrations for motility and biofilm formation by S. typhimurium. Furthermore, effectiveness of the CFCS against Salmonella was dose-dependent (P<0.05).          Conclusion: L. casei CFCS is able to inhibit the growth, motility and biofilm formation in S. typhimurium.          Keywords: Anti-Bacterial, Lactobacillus Casei Metabolites, Phenotypic Characteristics of S. typhimurium

Golestan University of Medical Sciences Medical Laboratory Journal 2538-4449 10 6 2016 11 1 Effect of Benzalkonium Chloride on Biofilm of Bacteria Causing Nosocomial Infectionstions 14 20 FA Azizollah Ebrahimi kahrizsangi Department of Pathobiology, Faculty of Veterinary Medicine Saied Habibian Dehkordi Department of Basic Sciences, Faculty of Veterinary Medicine Ziba Shabanpur Department of Bacteriology, Faculty of Veterinary Medicine Reza Hakimi Alni Department of Bacteriology, Faculty of Paraveterinary Majid Hemati Department of Bacteriology, Faculty of Veterinary Medicine ABSTRACT          Background and Objective: Biofilms are community of bacteria that attach to inanimate surfaces or living tissues via production of extracellular polymers and exopolysaccharide matrix. Microbial biofilms on various surfaces of the hospital environment are considered as a reservoir of infection spread. The present study aimed to evaluate the disinfecting effect of benzalkonium chloride on some bacterial isolates causing nosocomial infections.        Methods: First, 13 isolates from four bacteria including Pseudomonas aeruginosa, Staphylococcus aureus, Acinetobacter and Enterobacter were obtained from Microbiology Laboratory of Al-Zahra Hospital in Isfahan, Iran. The samples were transferred to Microbiology Laboratory of Faculty of Veterinary Medicine of Shahrekord University for testing. Evaluation of biofilm formation and determination of minimum inhibitory concentration (MIC) of the disinfectant and effect of the disinfectant on planktonic growth and biofilm formation were performed.         Results: All bacterial isolates (52 cases) produced biofilm. Mean MIC of benzalkonium chloride for P. aeruginosa, S. aureus, Enterobacter and Acinetobacter was 0.14, 0.2, 0.18, 0.17 g/ml, respectively. Planktonic growth of all four bacteria was inhibited at concentrations of 2MIC, MIC and 1/2MIC. Biofilm was not produced in MIC and 2MIC concentrations, and biofilm formation capability increased by reducing the concentration of benzalkonium chloride.           Conclusion: The results show that the use of appropriate concentration of benzalkonium chloride can prevent the growth of different bacterial species, but sub-MIC dose of this disinfectant may stimulate biofilm formation.             Keywords: Biofilm, Benzalkonium Chloride, Pseudomonas Aeruginosa, Staphylococcus Aureus, Enterobacter, Acinetobacter.

Golestan University of Medical Sciences Medical Laboratory Journal 2538-4449 10 6 2016 11 1 Effect of Mouse Liver Extract on in Vitro Differentiation of Amniotic Membrane Stem Cells into Hepatocyte-Like Cells 21 27 FA Vahide Vahideh assadollahi Department of Biochemistry and Genetic, Faculty of Medicine Masoume Jalalvand Department of Biotechnology, Faculty of Medicine Shahrokh Bagheri Department of Biochemistry and Genetic, Faculty of Medicine Hamed Esmaiel Lashkarian Department of Biochemistry and Genetic, Faculty of Medicine ABSTRACT           Background and Objective: Multipotent placental amniotic membrane mesenchymal stem cells (MSCs) are capable of differentiating into specialized tissues under different conditions. The aim of this study was to induce differentiation of placental amniotic membrane MSCs from NMRI mouse into hepatocytes using liver extract.          Methods: Placental amniotic membrane MSCs from a 14-day pregnant female mouse was used in this study. The cells were incubated with trypsin solution, followed by pipetting. The resulting suspension was cultured in 12-well plates. After confirming their mesenchymal nature, differentiation of the aforementioned cells was induced via exposure to 6, 18, 30 and 60 μg/ml of liver extract. On the 16th day of treatment, immunocytochemical reaction for albumin and periodic acid-Schiff (PAS) test were performed for detection of hepatocyte-like cells.           Results: Change was observed in the shape of differentiating cells from spindle-like shape to polygonal shape. The immunocytochemical reaction of the differentiated cells was positive. PAS staining also confirmed the accumulation of glycogen particles in the aforementioned cells. Concentration of 6 μg/ml liver extract was found as the effective dose for induction of differentiation.            Conclusion: The findings of this study show that the placental amniotic membrane-derived MSCs of mouse can differentiate in vitro from spindle-like cells to polygonal hepatocyte-like cells with large nuclei and under the influence of the liver. Keywords: Placental Amniotic Membrane Mesenchymal Stem Cells, Hepatocyte, In Vitro.

Golestan University of Medical Sciences Medical Laboratory Journal 2538-4449 10 6 2016 11 1 In vitro Susceptibility Study of Salmonella typhimurium Mutant to Stress Factors 28 31 FA Ania Ahani Azari Department of Microbiology, Gorgan Branch Taghi Zahraei Salehi Department of Microbiology, Faculty of Veterinary Medicine Bahar Nayeri Fasaei Department of Microbiology, Faculty of Veterinary Medicine ABSTRACT         Background and Objective: In this study, we compared the susceptibility of Salmonella typhimurium phoP mutant and its parent to stress conditions that the pathogen may encounter in a host.         Methods: For this purpose, we used the phoP deletion mutant constructed in our previous study. In order to test the in vitro susceptibility of the mutant to stress factors, the effect of acidic pH, heat, bile salts and polymyxin on growth of the mutant was examined. Then, minimum inhibitory concentration and minimum bactericidal concentration of bile salts and polymyxin were determined. Salmonella typhimurium 14028 was used as the parent strain.          Results: The mutant was highly susceptible to bile salts and polymyxin in comparison with the parent strain, but no difference was observed in their susceptibility to acid and heat.           Conclusion: This study confirms the role of the phoP in resistance of Salmonella to polymyxin and bile salts. Mutation in the phoP leads to susceptibility of the mutant to bile and cationic antimicrobial peptides.          Keywords: In vitro, Mutant, Salmonella typhimurium, Disease Susceptibility.

Golestan University of Medical Sciences Medical Laboratory Journal 2538-4449 10 6 2016 11 1 Effect of 8-week Aerobic Exercise on Undercarboxylated Osteocalcin and Beta Cell Function in Postmenopausal Women 32 37 FA Asieh Abbassi Daloii Department of Sport Physiology Islamic Azad University Esmatalsadat Mousavi Department of Sport Physiology Islamic Azad University ABSTRACT           Background and Objective: The present study aims at investigating the possible effect of 8-week aerobic exercise on undercarboxylated osteocalcin and beta cell function in postmenopausal women.          Methods: The study included 20 postmenopausal women with mean weight, height and body mass index of 78.94 ± 5.72 kg, age 48.69 ± 3.21 years, 160.37 ± 4.12 cm and 30.72 ± 2.37 kg/m2. The participants were randomly selected and divided into experimental and control groups. Blood samples were taken 48 hours before the experiment and after eight weeks of exercise. Aerobic exercise was performed for eight weeks, three sessions per week with intensity of 65-70% of heart rate. Data analysis for intragroup and intergroup differences was done using paired and independent t-test, respectively. P-values less than 0.05 were considered statistically significant.                      Results: The level of undercarboxylated osteocalcin in the experimental group decreased significantly compared to control group (P<0.049). The level of beta cell function index in the experimental group increased significantly after the 8-week exercise program compared to  control group (P<0.014).          Conclusion: Exercise increases the level of undercarboxylated osteocalcin in postmenopausal women that has important consequences, especially for those at risk of developing diabetes.             Keywords: Exercise, Osteocalcin, Hemoglobin A, Glycosylated, Postmenopause.

Golestan University of Medical Sciences Medical Laboratory Journal 2538-4449 10 6 2016 11 1 Production of Specific Egg Yolk Antibody (IgY) against Cryptosporidium parvum Oocysts 38 42 FA Hossein Hashemzadeh Farhang Department of Pathobiology, Tabriz Branch Parisa Shahbazi Department of Pathobiology, Faculty of Veterinary Medicine ABSTRACT          Background and Objectives: Cryptosporidium parvum is a coccidian protozoan that causes diarrhea in immunocompromised humans and newborn animals. The severity of the disease depends on the immunological status of the affected. Cryptosporiosis can have lethal effects on immunocompromised individuals such as AIDS patients. About 10% of AIDS patients die following an infection with C. parvum. Since there is no efficient treatment for cryptosporidiosis, there is an urgent need to search for more effective and safer alternatives. IgY is an avian immunoglobulin found in egg yolks. Due to its several advantages, IgY technology has been successfully used in biomedical research on humans and animals in the recent years. In this study, the specific chicken egg yolk antibody (IgY) against C. parvum whole oocyst antigens was produced and examined.          Methods: The effect of specific chicken egg yolk antibody (IgY) against whole oocyst antigens was examined. IgY sample was obtained from eggs of chickens immunized with C. parvum whole oocyst antigens and analyzed with lysate of C. parvum oocysts by dot blot assay.          Results: The IgY was produced with concentration of 9.7 mg/ml. This antibody was able to recognize the whole oocyst antigens until the dilution of 1:1000, but the best dilution for other immunoassays was 1:500.          Conclusion: Since chicken egg yolk is a cheap and convenient source for mass production of specific antibodies, the use of IgY against whole oocyst antigens could be considered a suitable candidate for passive immunization against cryptosporidiosis in humans and animals.          Keywords: Cryptosporidium parvum, IgY, Oocysts.

Golestan University of Medical Sciences Medical Laboratory Journal 2538-4449 10 6 2016 11 1 Effects of Citrus aurantium Fruit Extract on Spermatogenesis in Streptozotocin-Induced Diabetic Rats 43 48 FA Leili Komeilifard Department of Biology, Jahrom Beranch Vahid Hemayat khahjahromi Department of Biology, Jahrom Beranch ABSTRACT           Background and Objectives: Diabetes is one of the most common endocrine disorders, which is associated with changes in testicular tissue. The present study investigated therapeutic and prophylactic properties of bitter orange (Citrus aurantium) juice on testicular tissue and spermatogenesis process.           Methods: Forty streptozotocin-induced diabetic Wistar rats aged three months with mean weight of 170-200 g were divided into 4 groups including 1) control group, 2) diabetic control group, 3) diabetic group receiving 100mg/kg C. aurantium extract and 4) diabetic group receiving 200 mg/kg C. aurantium extract. The extract was administered to the rats for 56 days by gavage. After this period, the rats were anesthetized with ether and then their testes were fixed in 10% formalin for sample preparation. The testicular tissue was examined by haematoxylin and eosin staining under a light microscope with 10 and 40 magnifications. The mean number of Leydig and Sertoli cells, spermatogonia, spermatocytes and spermatids were calculated.            Results: A significant decrease was observed in mean weight of left testis in diabetic rats compared to that of controls (P≤0.05). The mean weight of testes showed no significant difference in diabetic rats treated with 200 mg/kg of extract compared with the control group. Diabetes reduced the number of spermatogonia, spermatocytes, spermatids and Sertoli cells. The number of cells increased significantly in the diabetic group receiving 200 mg/kg of extract. The spermatocytes and spermatids in both groups treated with the extract increased significantly.           Conclusion: This study shows the positive effect of bitter orange extract on complications of diabetes in testicular tissue. Therefore, this extract could be used for therapeutic purposes.            Keywords: diabetes, bitter orange juice, spermatogenesis, testis, rat

Golestan University of Medical Sciences Medical Laboratory Journal 2538-4449 10 6 2016 11 1 Evaluation of Antibacterial Effects of Aqueous and Alcoholic Extracts of Nasturtium Officinale on Some Pathogenic 49 53 FA Saeid Farshbaf Derhami Department of Microbiology, Faculty of Basic science Biology Mehdi Ghiami Rad Department of Microbiology, Faculty of Basic science Biology Razzagh Mahmoudi Department of Food Hygiene and Aquatics, Faculty of Veterinary Medicine ABSTRACT          Background and Objective: Although antibiotics are commonly used for treatment of infectious diseases, these treatments are often associated with several problems such as unwanted side effects and resistance to antibiotics. The aim of this study was to evaluate the antibacterial effect of aqueous and alcoholic extracts of Nasturtium officinale on Escherichia coli, Salmonella typhimurium, Staphylococcus aureus and Listeria monocytogenes.           Methods: All experiments were performed using the well diffusion method. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the aqueous and alcoholic extracts of N. officinale against the pathogenic bacteria studied were determined by microdilution method.          Results: In the well diffusion method, S. aureus and L. monocytogenes were the most sensitive bacteria with MIC of 8 μg/ml, while E. coli and S. typhimurium were the most resistant bacteria to the aqueous and alcoholic extracts. In addition, the inhibitory activity of the alcoholic and aqueous extracts of N. officinale was higher against gram-positive bacteria compared to gram-negatives. The lowest MIC (6.25 μg/ml) and MBC (12.5 μg/ml) of the plant extract were against S. aureus.           Conclusion: The aqueous and alcoholic extracts of N. officinale affect the growth of gram-positive bacteria (S. aureus and L. monocytogenes) but not the gram-negatives (S. typhimurium and E. coli). These extract could be used for treatment of infections.         Keywords: Nasturtium Officinale, Aqueous Extract, Alcoholic Extract, Antibacterial Effect.

Golestan University of Medical Sciences Medical Laboratory Journal 2538-4449 10 6 2016 11 1 Antibacterial Activity of Extract and Essential Oil of Ziziphora tenuior L. against Bacteria Isolated from Urogenital Tract Infections 54 59 FA Younes Anzabi Department of Pathobiology, Tabriz Branch Arash Khaki Department of Pathobiology, Tabriz Branch ABSTRACT          Background and Objectives:  Many species of  Ziziphora have been used in traditional medicine in the Azarbaijan region, northwest of Iran. This study aimed to determine antibacterial activity of essential oil and methanol extract of Ziziphora tenuior on some pathogenic bacteria isolated from urogenital tract infections.          Methods: The essential oil and methanol extract of Z. tenuior were obtained by Clevenger and maceration methods. Under sterile conditions, the required amount of urine was taken from patients referred to a number of clinics in Tabriz during 2014. After identification of the isolates using standard microbiological methods, antimicrobial effects of the essential oil and methanol extract of the plant on the isolates were evaluated by determining minimum inhibitory concentration (MIC), minimum bactericidal concentration, and antibiogram test.           Results: The MIC for essential oil of Z. tenuior was 250µg/mL for most Gram-negative bacteria except Pseudomonas aeruginosa. Moreover, the MIC for Staphylococcus aureus was 250µg/mL and 500µg/mL for other strains of Staphylococcus.            Conclusion: Comparison of the inhibitory and bactericidal effects of the essential oil and methanol extract of Z. tenuior showed that the essential oil is able to inhibit growth of the bacteria tested even in low concentrations. Further studies are required in this regard using animal models.          Keywords: Antibacterial Agents, Ziziphora tenuior, Plant Extracts, Urinary Tract Infection.