Resistance to Chlorhexidine and Benzalkonium Chloride in E. Coli ST131, A. Baumannii, and P. Aeruginosa Isolates.

BACKGROUND: The goal was to assess the antimicrobial efficacy of two commonly used biocides, chlorhexidine, and benzalkonium chloride, against MDR isolates of Pseudomonas aeruginosa, Acinetobacter baumannii, and Escherichia coli ST131, as well as the prevalence of resistance genes. METHODS: MIC of chlorhexidine and benzalkonium chloride and their effects on both the planktonic phase and biofilm were determined. Finally, the presence of genes responsible for resistance to quaternary ammonium compounds was investigated by PCR. RESULTS: No significant relationship was observed between the presence of resistance genes and different concentrations of quaternary ammonium compounds (benzalkonium chloride). There was no association between biofilm formation and the presence of resistance genes. CONCLUSIONS: Chlorhexidine digluconate and benzalkonium chloride at appropriate concentrations could prevent biofilm formation.

Virulence Genes and Biofilm Formation Among Legionella pneumophila Isolates Collected from Hospital Water Sources.

Legionella pneumophila can be transmitted to people, especially immunocompromised patients, via hospital water pipe systems and cause severe pneumonia. The aim of our study was to investigate the presence of major virulence factor genes, ability of biofilms formation, and correlation between presence of Legionella isolates and temperature, pH, and residual chlorine of water. Hundred water samples were collected from nine hospitals in Tehran, Iran. Temperature, pH, and residual chlorine were determined during sampling. Different virulence genes and the ability to form biofilms were subsequently analyzed among the L. pneumophila isolates. Results showed that 12 (12%) samples were positive in culture method and all of the isolates were positive as L. pneumophila species (mip). A correlation was found between Legionella culture positivity and temperature and pH of water, but there was no significant correlation between residual chlorine of water samples and the presence of Legionella. The isolation of Legionella rate in summer and spring was higher than winter and autumn. Twelve (100%) isolates were positive for mip genes, 9 (75%) for dot genes, 8 (66.66%) for hsp, 6 (50%) for lvh, and 4 (33.33%) for rtx. All of the isolates displayed strong ability for biofilm production every three days. Two of these isolates (16.6%) displayed weak ability to form biofilm on the first day of incubation. This study revealed that water sources in hospitals were colonized by virulent Legionella and should be continuously monitored to avoid elevated concentrations of Legionella with visible biofilm formation.

Molecular determination of O25b/ST131 clone type among extended spectrum ß-lactamases production Escherichia coli recovering from urinary tract infection isolates.

BACKGROUND: Escherichia coli (E. coli) O25b/ST131 clone causes urinary tract infection (UTI) and is associated with a broad spectrum of other infections, such as intra-abdominal and soft tissue infections, that can be affecting bloodstream infections. Therefore, since O25b/ST131 has been reported in several studies from Iran, in the current study, we have investigated the molecular characteristics, typing, and biofilm formation of O25b/ST131 clone type E. coli collected from UTI specimens. METHODS: A total of 173 E. coli isolates from UTI were collected. The susceptibility to all fourth generations of cephalosporins (cefazolin, cefuroxime, ceftriaxone, cefotaxime, ceftazidime, cefepime) and ampicillin, ampicillin-sulbactam and aztreonam was determined. Class A ESBLs, class D ESBL and the presence of pabB gene screenings to detect of O25b/ST131 clone type were performed by using of PCR. Biofilm formation was compared between O25b/ST131 isolates and non-O25b/ST131 isolates. Finally, ERIC-PCR was used for typing of ESBL positive isolates. RESULTS: Ninety-four ESBL positive were detected of which 79 of them were O25b/ST131. Antimicrobial susceptibility test data showed that most antibiotics had a higher rate of resistance in isolates of the O25b/ST131 clonal type. Biofilm formation showed that there was a weak association between O25b/ST131 clone type isolates and the level of the biofilm formation. ERIC-PCR results showed that E. coli isolates were genetically diverse and classified into 14 groups. CONCLUSION: Our results demonstrated the importance and high prevalence of E. coli O25b/ST131 among UTI isolates with the ability to spread fast and disseminate antibiotic resistance genes.

Safety evaluation of mutagenicity, genotoxicity, and cytotoxicity of Lactobacillus spp. isolates as probiotic candidates.

BACKGROUND: Probiotics promote a healthy balance of gut bacteria and have many beneficial effects on human digestive physiology. Although, few side effects of probiotics have been reported. This study aimed to assess the safety of five probiotic candidate Lactobacillus strains isolated from healthy individuals by examining mutagenicity, genotoxicity, and oral toxic effects. METHODS: Five selected candidate probiotic (SCPs) strains were evaluated for genotoxicity (Ames test with Salmonella typhimurium), in vitro mammalian chromosome aberration test and an in vivo mouse micronucleus assay on peripheral blood of mice. To evaluate the oral dose toxicity, BALB/c mice models were treated repeatedly (2000, 1000, and 500 mg/kg body weight /day) for 28-days. RESULTS: The Ames test performed for two S. typhimurium strains TA 98 and TA100 (both in the absence and in the presence of S-9 metabolic activation system) did not show an increase in reverse mutation because of exposure to the SCPs in any of the doses (5.0, 2.5, 1.25, 0.625, and 0.3125 mg/plate). There was no genotoxicity in the SCPs treatment in the vitro chromosome aberration assay with Chinese hamster ovary cells (CHO-K1). In addition, none of the tested strains increased the frequency of micronucleated reticulocytes in reticulocytes, the SCPs with the studied doses caused no substantial variation in the experimental groups compared to the negative control group (p > 0.05). SCPs were not acutely toxic when administered to male and female BALB/c mice by single gavage at (2000, 1000, and 500 mg/kg b.w/day) with no mortality or clinical signs, change in body weight or macroscopic abnormalities were observed in this dose range. CONCLUSION: As a result, SCPs did not induce mutagenic potential in vitro with bacterial reverse mutation, clastogenicity, and in vivo tests in the ranges of concentrations evaluated in our study.

Distribution and expression of virulence genes (hlyA, sat) and genotyping of Escherichia coli O25b/ST131 by multi-locus variable number tandem repeat analysis in Tehran, Iran.

Escherichia coli ST131 is a pandemic clone with high antibiotic resistance, and it is a major causative agent of urinary tract infection (UTI) and bloodstream infections. This study evaluated the distribution and expression of virulence genes and genotyping of E. coli O25b/ST131 by Multi-locus variable number tandem repeat analysis (MLVA) method among UTI in patients at Tehran hospitals, Iran.A total of 107 E. coli isolates were collected from UTI patients. Polymerase chain reaction (PCR) amplification of the pabB gene was used to identify E. coli O25b/ST131 and the prevalence of sat and hlyA virulence genes was also analyzed. The microtiter method quantified biofilm formation ability in E. coli O25b/ST131. The Real-Time PCR (qRT-PCR) was performed to evaluate the expression of sat and hlyA genes. Finally, MLVA was performed for E. coli O25b/ST131 genotyping by targeting seven tandem repeats. SPSS-16 software was used for statistical analysis. Molecular study showed that 71% of isolates carried the pabB gene and were considered E. coli O25b/ST131 strains. Also, 45.8% and 17.8% of isolates carried sat and hlyA genes, respectively. The 57.9% isolates had biofilm formation ability. Expression of the studied virulence genes showed an increase in strong biofilm producing E. coli O25b/ST131 strains. A total of 76 (100%) E. coli O25b/ST131 strains were typed by the MLVA method.High prevalence of E. coli O25b/ST131 isolates in UTI patients can be a serious warning to the treatment due to the high antibiotic resistance rate, expression of virulence genes, and biofilm formation.

Assessment of the risk of Legionella pneumophila in water distribution systems in hospitals of Tehran city.

When a sensitive host inhales aerosols containing these bacteria, Legionella infection occurs. Therefore, monitoring and assessing Legionella in the environment and water distribution systems of such places are critical due to the prone population in hospitals. However, the health risks of Legionella bacteria in the environment are not adequately evaluated. In this study, for hospitalized patients, we performed a quantitative health risk assessment of Legionella in selected hospitals in Tehran city using two scenarios of shower and toilet faucet exposure. This study identified Legionella in 38 cases (38%) out of 100 samples collected from toilet faucets and showers in 8 hospitals. The information gathered was used for quantitative microbial risk assessment (QMRA). The microbial load transmitted by inhalation was calculated using the concentration of Legionella in water. Other exposure parameters (inhalation rate and exposure time) were obtained using information from other studies and the median length of hospital stay (3.6 days). The exponential model was used to estimate the risk of infection (γ = 0.06) due to Legionella pneumophila (L. pneumophila) inhalation for each exposure event. For the mean concentration obtained for Legionella (103 CFU/L), the risk of infection for toilet faucets and showers was in the range of 0.23-2.3 and 3.5-21.9, respectively, per 10,000 hospitalized patients. The results were compared with the tolerable risk level of infection determined by the US EPA and WHO. The risk values exceeded the WHO values for waterborne pathogens in hospitals in both exposure scenarios. As a result, our QMRA results based on monitoring data showed that despite using treated water (from distribution networks in the urban areas) by hospitals, 38% of the samples were contaminated with Legionella, and faucets and showers can be sources of Legionella transmission. Hence, to protect the health of hospitalized patients, the risk of Legionella infection should be considered.

Bacterial resistance to antimicrobial peptides.

Antimicrobial peptides (AMPs) or host defense peptides (HDPs) are vital components of human innate defense system targeting human-related bacteria. Many bacteria have various mechanisms interfering with AMP activity, causing resistance to AMPs. Since AMPs are considered as potential novel antimicrobial drugs, understanding the mechanisms of bacterial resistance to direct killing of AMPs is of great significance. In this review, a comparative overview of bacterial strategies for resistance to direct killing of various AMPs is presented. Such strategies include bacterial cell envelope modification, AMP degradation, sequestration, expelling, and capsule.

Virulence factors and antimicrobial resistance in uropathogenic Escherichiacoli strains isolated from cystitis and pyelonephritis

Background/aim: The aim of this study was to investigate the prevalence of virulence genes as well as patterns of antibiotic resistance in cystitis and pyelonephritis uropathogenic Escherichia coli (UPEC) isolates. Materials and methods: Two hundred UPEC isolates were collected from hospitalized patients with pyelonephritis (n = 50) and cystitis (n = 150) in Shafa Hospital in Iran. Antimicrobial susceptibility and ESBL production were determined with confirmatory tests. Polymerase chain reaction assay was performed to determine the prevalence of virulence genes in UPEC strains. Results: Of a total 200 UPEC isolates, the highest and lowest resistance rates to antibiotics were for cephalexin (74%) and nitrofurantoin (9%), respectively. Of these isolates, 72 (36%) and 128 (64%) strains were ESBL-positive and ESBL-negative, respectively. The frequency of fimH, papC, and hly was 64%, 38%, and 12%, respectively. The most commonly identified virulence gene in ESBL-positive and ESBL-negative strains was fimH 46 (23%) and 86 (43%), respectively. The hlyA gene was more prevalent among patients with pyelonephritis than cystitis. Conclusion: The frequency of virulence genes was not significantly different between pyelonephritis and cystitis UPEC strains in the studied patients, but the prevalence rates of hlyA and papC genes were higher among UPEC strains isolated from inpatients compared to outpatients; hence, they could be considered as useful targets for prophylactic interventions.

Determination of antimicotic susceptibility pattern of Candida species isolated from patients with symptomatic candiduria.

BACKGROUND: The present study was conducted to determine antimicotic susceptibility of Candida species (sp.) from patients with symptomatic candiduria. MATERIALS AND METHODS: Identification of Candida sp. and determination of efficacy of most routine antifungals were done using polymerase chain reaction-restriction fragment length polymorphism method and E-test, respectively. RESULTS: The results from susceptibility test reveal that caspofungin and amphotericin B have high antifungal activity against both albicans (100% and 96%, respectively) and nonalbicans (95.11% and 72.72%, respectively) isolates. CONCLUSION: The present study suggests that caspofungin and amphotericin B have the excellent ability to eradicate both Candida groups that showed decreased susceptibility to other compounds.

Anti-Vibriocholerae IgY Antibody Inhibits Mortality in Suckling Mice Model.

BACKGROUND: Regarding to the importance of cholera in Iran and the potential advantages of egg yolk antibody (IgY) for immunotherapy, the aim of this study was to produce IgY antibody against V. cholerae Lipopolysaccharide (LPS) and determine its potential for V. cholerae treatment. METHODS: LPS was prepared, and the Anti-V. cholerae LPS IgY was purified from egg yolk and serially diluted in phosphate-buffered saline (PBS), mixed with V. cholerae and then gavaged into several groups of suckling mice. RESULTS: The yield of Anti-LPS IgY extraction was 40 mg/Egg yolk. The results demonstrated that up to approximately 75 ng of IgY can detect specifically V. cholerae. The lowest protective dose of anti-V. cholerae LPS IgY was 2.5 µg. CONCLUSIONS: The produced anti-Vibrio LPS specific IgY showed a good reactivity with its specific antigen and it may use as a complimentary oral immunotherapy for cholera disease.

Brucella melitensis VirB12 recombinant protein is a potential marker for serodiagnosis of human brucellosis.

BACKGROUND: The numerous drawbacks of current serological tests for diagnosis of brucellosis which mainly results from cross reactivity with LPS from other gram-negative bacteria have generated an increasing interest to find more specific non-LPS antigens. Previous studies had indicated that Brucella VirB12 protein, a cell surface protein and component of type IV secretion system, induces antibody response during animal infection. However, this protein has not yet been tested as a serological diagnostic marker in human brucellosis. METHODS: Recombinant VirB12 protein was prepared and evaluated the efficacy of it in an indirect enzyme-linked immunosorbent assay (ELISA) for brucellosis with sera collected from different region of Iran and the results were compared with a commercial ELISA kit. RESULTS: Sera from human brucellosis patients strongly reacted to the purified recombinant VirB12. The sensitivity, specificity, accuracy, negative predictive value and positive predictive value of recombinant VirB12-based ELISA related to the commercial-ELISA method were 87.8, 94, 90, 80 and 96.6% respectively. CONCLUSIONS: We concluded that antigenic VirB12 have a property value that can be considered as a candidate for using in serodiagnostic tests for human brucellosis.

An overview of case reports and case series of pulmonary actinomycosis mimicking lung cancer: a scoping review.

Background: Pulmonary actinomycosis (PA) is a rare type of Actinomyces infection that can be challenging to diagnose since it often mimics lung cancer. Methods: Published case reports and case series of PA in patients with suspicion of lung cancer were considered, and data were extracted by a structured search through PubMed/Medline. Results: After analyzing Medline, 31 studies were reviewed, from which 48 cases were extracted. Europe had the highest prevalence of reported cases with 45.1%, followed by Asia (32.2%), America (19.3%), and Africa (3.2%). The average age of patients was 58.9 years, and 75% of all patients were above 50 years old. Male patients (70%) were predominantly affected by PA. The overall mortality rate was 6.25%. In only eight cases, the causative agent was reported, and Actinomyces odontolyticus was the most common isolated pathogen with three cases. Based on histopathological examination, 75% of the cases were diagnosed, and the lobectomy was performed in 10 cases, the most common surgical intervention. In 50% of the cases, the selective antibiotics were intravenous and oral penicillin, followed by amoxicillin (29.1%), amoxicillin-clavulanic acid, ampicillin, levofloxacin, and doxycycline. Conclusion: The non-specific symptoms resemble lung cancer, leading to confusion between PA and cancer in imaging scans. Radiological techniques are helpful but have limitations that can lead to unnecessary surgeries when confusing PA with lung cancer. Therefore, it is important to raise awareness about the signs and symptoms of PA and lung cancer to prevent undesirable complications and ensure appropriate treatment measures are taken.

Efficacy and treatment outcome of infected patients with pulmonary Mycobacterium kansasii: A systematic review.

Background: Mycobacterium kansasii (M. kansasii) is a non-tuberculosis bacterium with a highly prevalent that is transferred by aerosols from water and soil resources to the respiratory system. M. kansasii is one of the main species responsible for NTM pulmonary disease. Methods: Web of Science, Scopus, and PubMed databases were systematically explored. Relevant articles from 1971 to November 2023 were reviewed. "The inclusion criteria" included patients with M. kansasii infection, treatment follow-up, and treatment outcomes. "The exclusion criteria" were clinical samples from animals, environmental samples, and other laboratory investigations. Results: 40 studies, including 1201 patients, were obtained through database search. Using the therapeutic regimens used in different studies, the therapy course for patients with M. kansasii infection ranged from 1 week to 118 months. In this study, the antibiotics prescribed in different treatment regimens for M. kansasii pulmonary infection were as follows: Rifampin, Ethambutol, Isoniazid, Clarithromycin, Streptomycin, and Pyrazinamide. Antibiotic combinations of three or four medicines, including rifampin, ethambutol, and isoniazid with or without streptomycin or pyrazinamide had the most therapeutic effect. Conclusion: The initial treatment involves rifampin, ethambutol, isoniazid, and pyridoxine, per the guidelines from the American Thoracic Society (ATS) and the Infectious Diseases Society of America (IDSA). Understanding the treatment plan and its outcomes is crucial for managing and determining the most effective therapy approach.

Investigating the antimicrobial and anti-inflammatory effects of Lactobacillus and Bifidobacterium spp. on cariogenic and periodontitis pathogens.

Background: The use of probiotics is emerging as an innovative approach to managing oral health issues and mediating the immune system. The current study assessed the in vitro impacts of non-orally isolated probiotics on periodontitis and tooth decay pathogens. Methods: Briefly, the persistence of probiotics in exposure to oral cavity enzymes, hydrogen peroxide, and saliva samples was examined. It was also investigated the biofilm formation and aggregation ability of probiotics, the adherence of probiotics in human gingival fibroblast cell (HGFC) lines and molar teeth samples, and the potential of probiotics to co-aggregate with oral pathogens. Additionally, the current study evaluated the effects of live probiotics on virulence gene expression, biofilm production of main oral pathogens, and changes in inflammation markers. Results: The probiotics remained alive when exposed to enzymes in the oral cavity, hydrogen peroxide, and saliva at baseline, 1, 3, and 5 h after incubation at 37°C (p-value <0.05). Probiotics demonstrated to produce biofilm and aggregation, as well as adherence to HGFCs and maxillary molars (p-value >0.05). They showed significant co-aggregation with oral pathogens, which were recorded as 65.57% for B. bifidum 1001 with S. mutans, 50.06% for B. bifidum 1005 with P. gingivalis, 35.6% for L. plantarum 156 with F. nucleatum, and 18.7% for B. longum 1044 with A. actinomycetemcomitans after 8 h of incubation. A balance between pro-inflammatory and anti-inflammatory cytokines, along with inhibition of biofilm formation and changes in virulence gene transcripts, were observed. However, most of these changes were not statistically significant (p-value >0.05). Conclusion: This study demonstrated the direct link between adhesiveness, aggregation, and biofilm formation with probiotic antibacterial activity. In addition to the careful selection of suitable probiotic strains, the concentration and origin of probiotic isolates should be considered.

Exploring the interplay between Fusobacterium nucleatum with the expression of microRNA, and inflammatory mediators in colorectal cancer.

Background: Fusobacterium nucleatum has been recognized as an important key bacterium in the cause and spread of colorectal carcinogenesis. Nevertheless, the clinical relevance of F. nucleatum in colorectal cancer (CRC) and its effect on immune factors and the tumor microenvironment have not been fully elucidated. Materials and methods: The frequency of F. nucleatum was measured in 100 paired tumor and normal tissue specimens by TaqMan quantification Real-Time Polymerase Chain Reaction (qPCR). The mRNA expression levels of cytokines (IL-6, IL-10, IL-12ß, IL-17, TNF-α, TLR-2, and TLR-4), and miRNAs (miR-21, miR-31) were examined. Eventually, any potential correlations between the molecular and clinicopathological features of the neoplastic samples and the abundance of F. nucleatum were analyzed. Results: The relative frequency of F. nucleatum was significantly increased in cancerous tissue compared to adjacent non-tumor tissues. Furthermore, the high level of F. nucleatum was significantly associated with histological grade III and IV CRC tissues (P = 0.027 and P = 0.022, respectively) and perineural invasion-positive patients (P = 0.037). In addition, the expression levels of IL-6, IL-17, TNF-α,IL-12ß, TLR-2, and TLR-4 as well as miR-21 and miR-31 showed a significant increase in the cancer group. A notable correlation was also observed between the high status of F. nucleatum and the expression of IL-6, TNF-α and miR-21. Conclusion: Our results emphasize the importance of F. nucleatum and changes in the expression of genes involved in CRC. Studying the microbial profile and gene expression changes in CRC patients may be a promising approach to improve screening methods and provide therapeutic strategies.

Evaluation of enterotoxigenic Bacteroides fragilis correlation with the expression of cellular signaling pathway genes in Iranian patients with colorectal cancer.

BACKGROUND: Colorectal cancer (CRC) is one of the most common cancers all over the world, and dysbiosis in the gut microbiota may play a role in colorectal carcinogenesis. Bacteroides fragilis can lead to tumorigenesis by changing signaling pathways, including the WNT/ß-catenin pathway. Therefore, in the present study, we investigated the correlation between the enterotoxigenic B. fragilis amount and the expression of signaling pathway genes involved in CRC. MATERIALS AND METHODS: B. fragilis was determined in 30 tumors and adjacent healthy tissues by the qPCR method. Next, the relationship between enterotoxigenic B. fragilis and the expression of signaling pathway genes, including CCND1, TP53, BCL2, BAX, WNT, TCF, AXIN, APC, and CTNNB1 was investigated. Additionally, possible correlations between clinicopathological features of the tumor samples and the abundance of B. fragilis were analyzed. RESULTS: The results showed that B. fragilis was detected in 100% of tumor samples and 86% of healthy tissues. Additionally, enterotoxigenic B. fragilis colonized 47% of all samples, and bft-1 toxin was the most frequently found isotype among the samples. The analysis showed that the high level of B. fragilis has a significant relationship with the high expression of AXIN, CTNNB1, and BCL2 genes. On the other hand, our results did not show any possible correlation between this bacterium and the clinicopathological features of the tumor sample. CONCLUSION: B. fragilis had a higher abundance in the tumor samples than in healthy tissues, and this bacterium may lead to CRC by making changes in cellular signaling pathways and genes. Therefore, to better understand the physiological effects of B. fragilis on the inflammatory response and CRC, future research should focus on dissecting the molecular mechanisms by which this bacterium regulates cellular signaling pathways.

Investigation of drug regimens and treatment outcome in patients with Mycobacterium Simiae: a systematic review.

INTRODUCTION: Mycobacterium simiae (M.simiae), a non-tuberculous mycobacterium (NTM), rare causes infection including localized pulmonary to disseminated disease in immunocompromised patients. An optimal pharmacological management practice has not yet been defined for this infection. This study investigates drug regimens and treatment outcomes in patients with M. simiae to describe different drug regimen with the therapeutic response. AREAS COVERED: The three databases PubMed, Scopus, and Web of science were systematically searched from June 1994 to June 2021 to retrieve relevant articles. The inclusion criterion included studies, which reported treatment outcomes in patients with M. simiae infections. Treatment success was defined as the achievement of culture conversion, and the improvement of the symptoms and radiologic signs among the patients. EXPERT OPINION: Data of 223 patients were retrieved from 40 studies. Duration of the treatment regimens used in different studies ranged from 2 to 12 months. The most common treatment regimens administered for M. simiae infection were as follows: clarithromycin, rifampin, ethambutol, moxifloxacin, or ciprofloxacin and amikacin plus cotrimoxazole or pyrazinamide in some regimens. Macrolides, such as clarithromycin, combined with quinolones (such as moxifloxacin) and TMP/SMX, which are used in combination, had the most significant effect on eliminating the pulmonary signs of M. simiae.

Potential of Lactobacillus acidophilus to modulate cytokine production by peripheral blood monocytes in patients with endometriosis.

Background and Objectives: Endometriosis is defined as the presence of endometrial tissue outside the uterine cavity. Peripheral blood monocytes cells (PBMCs) may have altered function to some extent in women with endometriosis. Lactobacillus acidophilus is a probiotic bacterium within the human body with the ability of alleviating many inflammatory diseases. Here, we examined the effect of L. acidophilus on PBMCs of endometriosis patients. Materials and Methods: In this study, peripheral blood samples were obtained from endometriosis patients (n=11) and non-endometriosis individuals (n=11). After isolation of peripheral blood mononuclear cells with Ficoll, cells were cultured in the presence and absence of phytohemagglutinin. Also, these cells were co-cultured with 1×106 CFU/ml of L. acidophilus. IL-6 and IL-1 cytokines were measured by ELISA method and the two groups were evaluated and compared. Results: The results showed that in endometriosis patients, the production of pro-inflammatory cytokines, including IL-1 and IL-6, by PBMC was increased compared to non-endometriosis subjects, and stimuli such as PHA intensified this elevation. Also, L. acidophilus increased the levels of pro-inflammatory cytokines including IL-1 and IL-6. However, the production of these cytokines decreased due to the modulatory properties of bacterial cells after 48 h. Conclusion: According to the results of the current study, IL-1 and IL-6 production was significantly increased in PMBCs of endometriosis patients compared to that of the healthy controls. Also, Lactobacillus acidophilus was considered as an antigenic compound and induced IL-1 and IL-6 production. According to these results, probiotics can be further used for the treatment of endometriosis patients and more investigations are needed to confirm these results.

Genotyping of Listeria monocytogenes isolates by high-resolution melting curve (HRM) analysis of tandem repeat locus.

Listeria monocytogenes is responsible for causing listeriosis, a type of food poisoning with high mortality. This bacterium is mainly transmitted to humans through the consumption of contaminated foods. Detection of L. monocytogenes through molecular methods is crucial for food safety and clinical diagnosis. Present techniques are characterized by low discrimination power and high cost, as well as being time-consuming and taking several days to give the final result. In our study, MLVA-HRM (Multiple-Locus Variable-number tandem repeats Analysis ‒ High-Resolution Melting) was investigated as an alternative method for a fast and precise method for the genotyping of L. monocytogenes isolates. Forty-eight isolates of L. monocytogenes obtained from the microbial bank of Department of Microbiology, Iran University of Medical Sciences, were typed by MLVA-HRM analysis using five Variable Numbers of Tandem Repeat (VNTR) loci. A total of 43 different types were obtained. This research demonstrated the usefulness of the MLVA-HRMA method and its ability to discriminate L. monocytogenes isolates. Since this method is easier and more efficient than existing methods, it can be widely used in food processing plants and diagnostic laboratories as a fast and accurate method.

Determination of investigation of the link between human and animal Brucella isolates in Iran using multiple-locus variable number tandem repeat method comprising 16 loci (MLVA-16).

BACKGROUND: Epidemiological studies are important tools to assess the diversity of Brucella isolates and to estimate their epidemiological relationship among isolates from different geographical origins. In this study the MLVA16 (multiple-locus variable number tandem repeat analysis based on 16 loci) was employed to investigate the diversity of Brucella spp. Isolated from humans and animals for epidemiological purposes and to determine the most common Brucella genotypes in Iran. METHODS: We designed a molecular-based study to evaluate the potential reservoirs of human brucellosis. After isolation and identification of 54 Brucella spp human and animal specimens from three regions of Iran, bacterial genomic DNA was extracted MLVA with three panel was used for the genotyping of isolates. The size of PCR products were analyzed and converted to repeat unit numbers using a published allele numbering system and data set was imported into Bionumerics. RESULTS: Three isolates (5.55%) were identified as Brucella abortus and 51 (94.44%) as Brucella melitensis. Two isolates of Brucella abortus were from humans and one from an animal. Thirty-four Brucella melitensis isolates were from humans and 17 from animals. Using MLVA16-genotyping, 54 isolates with genetic similarity coefficient of 80% were divided into 46 genotypes and 22 genotypes were represented by a single isolate, while 4, 2, 1 and 2 genotypes were represented by 2, 3, 4 and 7 isolates, respectively. The most prevalent genotype was represented by 14 isolates. There were two other frequent genotypes each represented by seven isolates, among which only one was restricted to a geographic region. Discriminatory power for each locus was determined in this study and panel 2B shows the high discretionary power [Bruce04 (0.837), Bruce30 (0.806), Bruce 09 (0.787), Bruce 07 (0.772), Bruce16 (0.766)]. CONCLUSION: MLVA16 analysis of 54 Brucella isolates showed high level polymorphism in their genotypes. Only two genotypes, each observed in seven isolates, were related to one another and only one of these genotypes were found in to two separate regions.