Assessment of gastric caused by Helicobacter pylori and pathologic elements correlation with -511 IL1-β and -308 TNF-α polymorphisms in gastritis patients

Helicobacter pylori (H.pylori) is the main reason for gastric disorders including gastric lymphoma, ulcer disease, gastric carcinoma (GC), and chronic atrophic gastritis. H.pylori has two more significant virulence factors named cagA and vacA. Some host cytokines polymorphisms (Interleukin (IL-1) and tumor necrosis factor (TNF-α)) may contribute to H. pylori-related diseases. In the present study, we investigated the association of H. pylori gastritis and its pathogenic genes as well as the association of IL-1β and TNF-α polymorphisms in patients with gastritis. We collected gastric biopsy samples from patients with gastritis. After extracting DNA from biopsy specimens infected with H. pylori, cagA + and vacA + were detected by the polymerase chain reaction (PCR). To genotyping TNF-α polymorphism at position − 308 and IL-1β polymorphism at position − 511, PCR-based restriction fragment length polymorphism analysis was performed. Our study indicated that IL-1β-511 polymorphism, unlike TNF-α-308 polymorphism (P = 0.030), did not show a significant relationship between patients infected with H. pylori (p = 0.219). Also, our results indicated that alleles C and T of polymorphism of IL-1β-511 and alleles G of TNFα-308 were not significantly correlated with cagA status among patients infected with H. pylori (p = 0.793, p = 0.674, p = 0.179, respectively) unlike allele A of TNFα − 308 (p = 0.016

Evaluation of antibacterial activity of fve biocides and the synergistic efect of biocide/ EDTA combinations on bioflm-producing and non-producing Stenotrophomonas maltophilia strains isolated from clinical specimens in Iran

Background: The overuse of biocides in healthcare-facilities poses risk for emergence and spread of antibiotic resistance among nosocomial pathogens. Hospital-acquired infections due to S. maltophilia have been increased in the recent years and with its various resistance mechanisms contribute to patient morbidity and mortality in hospitals. The current study aimed to evaluate the susceptibility of biofilm-producing and non-producing S. maltophilia clinical isolates to five commonly used hospital biocides, alone and in combination with EDTA to examine the synergistic effect of combining EDTA on the bactericidal activity of them by microbroth dilution method. As well as the frequency of efflux genes encoding resistance to biocides among isolates. This study also intended to assess the effect of exposure of S. maltophilia isolates to sub-inhibitory concentrations of sodium hypochlorite upon the antimicrobial susceptibility patterns. Results: Based on minimum inhibitory and bactericidal concentrations of biocides sodium hypochlorite 5% (w/v) and ethyl alcohol 70% (v/v) were the strongest and weakest biocides against S. maltophilia isolates, respectively. The combination of EDTA with biocides significantly increased the effectiveness of the studied biocides. Exposure to subinhibitory concentration of sodium hypochlorite showed a significant change in the susceptibility of isolates towards ceftazidime (p = 0.019), ticarcillin/clavulanate (p = 0.009), and chloramphenicol (p = 0.028). As well as among the isolates examined, 94 (95%) were able to produce biofilm. The frequency of sugE1 resistance genes was found in 90.7% of our clinical S. maltophilia isolates. None of the isolates carried qacE and qacEΔ1 gene. Conclusions: The current study recommended that using the mixture of biocides with EDTA can be effective in reducing nosocomial infections. Also, this study demonstrated that exposure to sub-inhibitory concentrations of sodium hypochlorite leads to reduced antibiotic susceptibility and development of multidrug-resistant S. maltophilia strains

Molecular characterization of Hymenolepis nana based on nuclear rDNA ITS2 gene marker

Introduction: Hymenolepis nana is a zoonotic tapeworm with widespread distribution. The goal of the present study was to identify the parasite in the specimens collected from NorthWestern regions of Iran using PCR-sequencing method. Methods: A total of 1521 stool samples were collected from the study individuals. Initially, the identification of hymenolepis nana was confirmed by parasitological method including direct wet-mount and formalin-ethyl acetate concentration methods. Afterward, PCR-sequencing analysis of ribosomal ITS2 fragment was targeted to investigate the molecular identification of the parasite. Results: Overall, 0.65% (10/1521) of the isolates were contaminated with H. nana in formalin-ethyl acetate concentration. All ten isolates were succefully amplified by PCR and further sequenced. The determined sequences were deposited in GenBank under the accession numbers MH337810 -MH337819. Conclusion: Our results clarified the presence of H. nana among the patients in the study areas. In addition, the molecular technique could be accessible when the human eggs are the only sources available to identify and diagnose the parasite. DOI: https://dx.doi.org/10.4314/ahs.v19i1.6 Cite as: Shahnazi M, Mehrizi MZ, Alizadeh SA, Heydarian P, Saraei M, Alipour M, Hajialilo E. Molecular characterization of Hymenolepis nana (Cestoda: Cyclophyllidea: Hymenolepididae) based on nuclear rDNA ITS2 gene marker. Afri Health Sci. 2019;19(1): 1346- 1352. https://dx.doi.org/10.4314/ahs.v19i1.

Molecular characterization of fasciola and dicrocoelium species isolated from ruminant livestock in Qazvin, Iran

Introduction: Fascioliasis and dicrocoeliasis are the most frequent zoonotic diseases with increasing human health problems in different parts of Iran. Two species, Fasciola hepatica (F. hepatica) and Fasciola gigantica (F. gigantica), are spread in the country. Molecular approaches have a decisive role in identifying both the species. The aim of this study was to detect Fasciola spp. and Dicrocoelium spp. by amplifying the ITS-2 and 28S rDNA gene sequence. Methods: Overall, 30 infected liver samples were collected from the livestock of Qazvin, Iran. The adult flukes were collected from different livestock. DNA extraction and PCR amplification of ribosomal RNA gene region (ITS2) and 28S rDNA gene fragment were conducted and a phylogenetic tree was constructed. Result: All the isolates obtained from the cattle (No: 7) and 82.6% (No: 19) of sheep isolates were infected with F. hepatica species, whereas 17.4% (No: 4) of sheep isolates were infected with F. gigantica. It was also shown that F. hepatica was the predominant species of Fasciola present in the region. All the specimens were infected with Dicrocoelium dendriticum (D. dendriticum). Conclusion: Both the species of Fasciola were found in Qazvin. D. dendriticum was the sole infecting species of the Dicrocoelium genus in the livestock of the city of Qazvin. Further research studies are needed to determine the intermediate host of the parasites in the region

Detection of bacterial agents causing prostate infection by culture and molecular methods from biopsy specimens

Background and Objectives: Prostatitis affects about 16% of men in their lifetime and sometimes leading to prostate cancer. Bacterial infections are the most common causes of prostatitis. Diagnosis of the causative agents of bacterial prostate infections plays an essential role in timely treating and preventing secondary complications. This study isolated bacterial infectious agents in patients’ surgical prostate and evaluated them by routine and molecular microbiological methods. Materials and Methods: In this cross-sectional study, 72 prostate biopsy specimens were collected from the Orology Departmen of hospitals of Qazvin University of Medical Sciences. All samples were cultured in aerobic and anaerobic conditions. Antibiotic susceptibility test by Kirby-Bauer standard method was performed for all isolated bacteria. In addition, all isolated bacteria were identified using 16S rDNA PCR and sanger sequencing methods. Also, TaqMan real-time PCR was applied to detect Ureaplasm aurealyticum, Mycoplasma hominins, and Mycoplasma genitalium. Results: In conventional culture method, out of 18 positive samples, 15 samples (83.3%) were Gram-negative bacteria and 3 samples (16.6%) were Gram-positive bacteria, containing Escherichia coli (55.5%), Klebsiella pneumoniae (11.1%), Enterobacter cloacae (5.5%), Pseudomonas aeruginosa (11.1%), Staphylococcus aureus (11.1%), and Enterococcus faecalis (5.5%). The results of molecular identification methods were the same as conventional culture results. Also, four patients were Ureaplasm aurealyticum, and three patients were positive for Mycoplasma hominis. Conclusion: Most bacteria isolated from prostate specimens belonged to the Enterobacteriaceae family, especially Escherichia coli, Klebsiella pneumoniae, Enterobacter cloacae. Staphylococcus aureus and Enterococcus faecalis were cocci isolated in the specimens too. Also, Ureaplasma urealyticum, and Mycoplasma hominis were identified in prostatitis. Keywords: Prostatitis; Pathogens; Enterobacteriaceae; 16s rDNA; Real-time polymerase chain reactio

Distribution and molecular analysis of Blastocystis subtypes from gastrointestinal symptomatic and asymptomatic patients in Iran

Introduction: Blastocystis is a common intestinal parasite of human and animal hosts. The parasite has 17 subtypes, and among those at least nine subtypes (ST1-ST9) are found in human hosts. Objective: The aim of the present study was to investigate the presence of different subtypes of Blastocystis spp. among the patients referred to Velayat hospital of Qazvin province, Iran. Methods: Overall, 864 stool samples were examined by using formalin-ethyl acetate concentration method and Trichrome staining. All specimens were cultured in clotted fetal bovine medium. Later, DNA extraction and PCR amplification of 18S ribosomal RNA gene region was conducted and phylogenetic tree constructed. Results: The results revealed 7.9% (68/864) of the study population were infected with Blastocystis. Intestinal symptoms were observed in 61% (36/59) of individuals positive for Blastocystis, with abdominal pain in 58% (21/36) of cases which was more frequent than other intestinal signs. No significant relationship was observed among the study variables. By molecular and phylogenetic analysis, three subtypes ST1 (45%), ST2 (30%) and ST3 (23%) of parasite were identified. Conclusion: This study showed ST1 subtype was the predominant subtype among the positive specimens, meanwhile the highest haplotype and nucleotide diversity were clarified in ST3 subtype

Screening and molecular characterization of Trichomonas vaginalis genotypes isolated from married women in northern Iran

. Trichomonas vaginalis is an anaerobic protozoan parasite that causes trichomonosis in human. It is one of the most common non-viral sexually transmitted infections. It has been found to be most prevalent in patients referred to sexually transmitted disease clinics. In recent years, molecular methods have been used to identify genotypes of this parasite in different parts of the world and so far 6 types of T. vaginalis have identified. The aim of this study was to investigate the prevalence and genotype identification of T. vaginalis from married women in northern Iran. A total of 450 vaginal specimens were taken from married women, referring to health centers in northern Iran. Demographic information of women was collected through a questionnaire. The samples were first examined microscopically and then monitored in Dorsch culture medium for up to 10 days. Actin genes of positive samples were amplified by PCR. Finally, PCR products were used to determine the sequence and genotype of the parasite. Overall, 0.7% (3/450) samples were positive for T. vaginalis. All of the three infected women were housewives. After sequencing, the genotype of these parasites were type H (66.7%) (Accession no; MW414672-MW414673) and type E (33.3%) (Accession no; MW414671). Low prevalence of T. vaginalis in north of Iran indicate high level of hygiene in sexual intercourse and avoiding from high risk sexual behaviors, and also it seems that genotype H is dominant type of the parasite in the study area

Phenotype and genetic determination of resistance to common disinfectants among bioflm-producing and non-producing Pseudomonas aeruginosa strains from clinical specimens in Iran

Background: Pseudomonas aeruginosa is a common pathogen in Hospitalized patients, and its various resistance mechanisms contribute to patient morbidity and mortality. The main aims of the present study were to assess the susceptibility of bioflm-producing and non-producing P. aeruginosa isolates to the fve commonly used Hospital disinfectants, to evaluate the synergistic efect of selected disinfectants and Ethylene-diamine-tetra acetic acid (EDTA), and the efect of exposure to sub-inhibitory concentrations of Sodium hypochlorite on antimicrobial susceptibility test. Results: The results showed that sodium hypochlorite 5% and Ethanol 70% were the most and least efective disinfectants against P. aeruginosa, respectively. The addition of EDTA signifcantly increased the efectiveness of the selected disinfectants. The changes in the antibiotic-resistance profles after exposure to sub-inhibitory concentrations of disinfectants were observed for diferent classes of antibiotics (Carbapenems, Aminoglycosides, Cephalosporins, Fluoroquinolones). As well as near the all isolates harbored efux pump genes and 117 (97.5%) of isolates produced bioflm. Conclusion: In the current study, the mixture of disinfectant and EDTA were the most suitable selection to disinfect Hospital surfaces and instruments. Also, it was clear that exposure to sub-inhibitory concentrations of Sodium hypochlorite results in resistance to some antibiotics in P. aeruginosa species. Strong and intermediate bioflm formers belonged to MDR/XDR strains. Future studies should include more complex microbial communities residing in the Hospitals, and more disinfectants use in Hospitals. Keywords: Nosocomial infection, Disinfectant-resistance, Bioflm, Hospital disinfectants, Pseudomonas aeruginosa, Clinical isolate

Simultaneous Detection of Pathogenic and Saprophyte Leptospira in Human Plasma by Multiplex Taqman Real Time PCR

Background: Leptospirosis is an infectious disease caused by pathogenic and saprophytic Leptospira species. The clinical and laboratory diagnosis of this infection is complicated. However, timely diagnosis of leptospirosis is essential for treatment of this disease. Conventional laboratory methods are incapable in the early diagnosis of it. Molecular tests such as real time PCR are very efficient when diagnosing it. Objectives: In this study, we designed and developed a multiplex Taqman real time PCR to simultaneously detect saprophyte and pathologic Leptospira in clinical samples. Methods: 250 human plasma samples were obtained from suspected patients. Two pair specific primers and the corresponding probe for detecting pathogenic and saprophytic Leptospira were designed and established in a single tube. The developed tests were run on all DNA extracted from the samples. Results: Of the 250 samples, 93 (37.2%) werepositive for pathogenicand 15 (6%) for non-pathogenic cases. In two samples, pathogenic and non-pathogenic DNA strains were simultaneously positive. Conclusions: Based on our finds, the real time PCR is a suitable test for the diagnosis of leptospirosis and differentiation between pathogen and saprophyte Leptospira simultaneously. Keywords: Leptospira, Real Time PCR, Saprophyte, Leptospirosi