Iranian Journal of Microbiology
Volume:11 Issue: 3, Jun 2019

Iron is an essential compound in metabolic pathway of wide range of organisms. Because of limited free iron supply in mammalian and avian hosts, bacteria have applied various ways to acquire iron. In this study, the frequency of 8 iron acquisition factors was examined among 63 avian and ovine Pasteurella multocida field isolates and their vaccine strains using PCR method. Five candidate genes (fur, tonB, exbD, exbB and hgbA) were identified among all isolates. For the first time, 2 loci (hgbB1 and hgbB2) of the hgbB gene were identified, which were previously reported as 1 gene. Also, it was found that 5 ovine and 1 avian isolates possessed all the virulence factors, which could also be considered for evaluating the frequency of other virulence factors. More studies need to be conducted on the frequency of all other virulence factors among these isolates, which can provide basic information for improvement or substitution of current vaccinal strains. Overall, as the new designed sets of primers showed more potential in detecting the corresponded genes, researchers can consider them in further studies.

Helicobacter pylori is a Gram-negative spiral-shaped bacterium that contaminates more than half of the world's inhabitants, and infection with this bacterium is associated with some gastric disorders. Also, 5% to 10% of H. pylori genes are specific to this bacterium and many bacterial virulence factors fall into this group. The cagA, vacA, sodB and hsp60 are among important virulence factors of H. pylori. A gastric biopsy specimen was taken from 341 gastric patients and cultivated on a Colombia agar plate, containing various antibiotics, such as vancomycin, amphotericin B, and trimethoprim & polymyxin B, and incubated for 3 to 10 days under microaerophilic conditions at 37°C. PCR was used to detect the ureC, cagA, vacA, sodB and hsp60 genes. In this study, 131 isolates were identified as H. pylori. The prevalence of cagA, vacA, sodB and hsp60 were 74%, 100%, 92.4% and 96.2%, respectively. The correlation between the clinical forms of the disease and the virulence genes were analyzed by statistical tests and no significant correlation was found.

The obtained results are similar to some studies conducted in different parts of the world and is different in other cases. This discrepancy is due to the difference in the type of gastric disorders, sample size and methodology.

Various non-invasive diagnostic tests are available for the detection of Helicobacter pylori infection. The aim of this study was to compare the sensitivity and specificity of HpSA, salivary IgG, serum IgG, and serum IgM to those of endoscopic-biopsy as the gold standard for the diagnosis of H. pylori infection.

This is a cross-sectional study performed among pediatric patients at Dr. Soetomo General Hospital (Surabaya, Indonesia). Fecal, blood, and saliva samples were collected from all subjects. The results of the HpSA, salivary IgG, serum IgG, and serum IgM tests were compared to the results of endoscopic-biopsy as the gold standard. Of the 37 study participants, H. pylori infection was confirmed in 5 (13.33%) with serum IgG, 23 (63.33%) with serum IgM, 15 (40%) with HpSA, and 26 (70.97%) with salivary IgG. The salivary IgG enzyme-linked immunosorbent assay (ELISA) was the only diagnostic test with significantly different results, as compared to biopsy (p = 0.017). The results of this study showed that HpSA, salivary IgG, and serum IgG and IgM were not sufficient to replace endoscopic-biopsy as the gold standard for the diagnosis of H. pylori infection.

Cholera disease remains an important global health problem affecting 3-5 million subjects worldwide. Outer membrane vesicles (OMVs) have been found in a variety of Gram-negative bacteria and act as protective transport vesicles. The aim of this study was to evaluate Immune responses against Vibrio cholerae O1 El Tor clinical strain OMV and compare it with killed whole cell (KWC), complex of (KWC-OMV) as well as the internationally licensed oral cholera vaccine, Dukoral, in serum and intestinal secretions of mice. OMVs were prepared by using modified detergent-centrifugation procedure from V. cholerae O1 El Tor clinical strain from 2005 outbreak. The ultrastructure and content of OMVs were investigated via the Scanning Electron Microscopy (SEM) and SDS-PAGE analysis. Three doses of oral immunization were adjusted and total IgG and IgA in serum and intestinal secretion were measured by enzyme-linked immunosorbent assay (ELISA). Extracted OMVs from the V. cholerae were spherical vesicles with a size ranging from 10 to 300 nm. OMV-immunized mice showed an increased level of total IgG and IgA both in serum and intestinal secretion when compared to the negative controls. Also, there existed a higher level of secretory IgA than the total IgG, suggesting the most of protection against V. cholerae colonization provided by sIgA. Our findings revealed that oral immunization with V. cholerae OMVs might induce a long-term immunity, especially when administered in combination with KWC. This study tested the adjuvant activity of OMVs and may be useful in future nano vaccine research.

Escherichia coli is a common enteric pathogen of human and livevestock. Antibiotic resistance is the main concern of public health. The aim of this study was to detect this bacterium in stool samples of diarrheal patients and identify the phenotypic and genotypic characterizations of antibiotic-resistant isolates such as dfrA1, sul1, citm, tetA, qnr, aac(3)-IV in Shahrekord. Two hundred fifty diarrheal stool samples from patients were collected. Microbiological and biochemical examinations were done to detect E. coli. Phenotypic and genotypic antibiotic resistance of the isolates were determined using dick diffusion method and polymerase chain reaction (PCR), respectively. Among 114 E. coli isolates, the least resistance was for gentamicin (0%) and the most resistance was for trimethoprim (79.8%). The resistance to sulfamethoxazole, ciprofloxacin, ampicillin, and tetracycline were 71.05%, 10.5%, 52.63%, and 3.5% respectively. The results of PCR assay revealed that 10 isolates contain sul1, 49 isolates harbor citm, 8 isolates tetA, 36 isolates dfrA1, 11 isolates qnr genes but there was no isolate with aac(3)-IV gene. In comparison between phenotypic and genotypic of the isolates revealed that citm, tetA, dfrA1, qnr, sul1, aac(3)-IV genes covered 42.98%, 7.01%, 31.57%, 9.64%, 8.7%, 0% of the antibiotic resistance, respectively. Our results revealed that all isolates harbor one or more antibiotic resistance genes and that the PCR is a fast practical and appropriate method to determine the presence of antibiotic resistance genes.

Klebsiella pneumoniae is an important cause of serious nosocomial infections among Gram-negative bacteria. The aim of this study was evaluating the prevalence of VIM-1, VIM-2, and IMP-1 metallo-β-lactamase genes in clinical specimens at two teaching hospitals in Sanandaj, Kurdistan west of Iran. Four hundred different clinical specimens were collected from hospitalized patients or referred to hospitals from May 2013 to March 2014 in Sanandaj, Kurdistan, Iran. MBLs – producing K. pneumoniae detected by Double Disk Synergy Test. The MBL positive isolates were examined for the presence of VIM-1, VIM-2 and IMP-1 genes using PCR technique. Of four hundred clinical specimens, 114 K. pneumoniae isolates were identified. Twenty-eight (24.6%) isolates were resistant to imipenem and 15 strains (53.6%) were positive for MBL enzymes production. PCR results showed VIM-1 and IMP-1 genes frequencies are 4 (26.7%) and 1 (6.7%). Only one strain of K. pneumoniae was found to be MBL producer among the outpatients. The study results exhibited a high level of resistance to most of the antibiotics tested and high prevalence of MBLs producing in K. pneumoniae at two hospitals. Thus, the infection control methods and the implementation of antibiotic agents should be taken into account.

Burkholderia mallei is the leading cause of glanders, a highly transmittable and an OIE-notifiable disease of equidae. Despite the importance of B. mallei, little is known about serodiagnosis of glanders. The present study aimed to develop an immunoblotting assay based on whole-cell proteome of B. mallei to enable accurate serodiagnosis of glanders. Three farm horses were subcutaneously immunized with a crude suspension (106 cfu/ml) of heat-inactivated B. mallei formulated with incomplete Freund's adjuvant (IFA) to achieve a hyperimmune sera panel. The immunization was done for 1, 14 and 28 days with 1 dose of 1 ml antigen containing 106 cfu/ml. The hyperimmunity of sera was confirmed by CFT. B. mallei whole-cell proteome was prepared through sonication and the protein content was visualized by SDS-PAGE and quantified by Western blot using HRP-conjugated rabbit anti-horse IgG. A comprehensive set of positive and negative horse sera validated the test. A ladder pattern of the B. mallei immunoreactive antigens was seen within the region of 20-90 kDa clearly and the immunoblot was scored positive, while no reaction was seen for the negative sera. The Western blot assay indicated a noticeably higher diagnostic specificity for positive or negative sera of glanders.


The whole-cell proteome-based immunoblot proved reliable and straightforward in our study. The prepared antigen was adaptable for application in immunoblotting. We assumed this improved immunoblotting system provides appropriate sensitivity and also specificity expected in serodiagnosis of glanders in endemic areas and typically in less-developed countries.

Although zinc oxide (ZO)-calcium hydroxide (CaOH) mixtures have been successful regarding their absorption rate compatibility with dissolving primary teeth, no study has been conducted on the appropriate mixture ratio to obtain effective antibacterial properties. In this study, we compared antibacterial activity of CaOH-ZO pastes using different mixture ratios sagainst Enterococcus faecalis as an important bacterium in root canal treatment failure. Seven types of pastes were prepared in our laboratory. The first group included one gram of ZO+eugenol, second group included one gram of CaOH+distilled water, third group included 0.5gram ZO+0.5gram CaOH+distilled water (1:1), forth group included 0.75gramCaOH+0.25gramZO+distilled water (3:1), the fifth group included 0.33gram of CaOH+0.66gram of ZO+distilled water (1:2), the sixth group included 0.75gram of ZO+0.25 CaOH+distilled water (3:1), the seventh group included 0.66 gram CaOH+0.33 gram ZO+distilled water (2:1), and the final group included one gram of gelatin+distilled water (as the control group). These pastes were compared regarding their antibacterial effects against Enterococcus faecalis using agar diffusion and microdilution methods. Except for the control group, all prepared pastes showed antibacterial properties. Order of minimum inhibitory concentration for pastes were as followed: CaOH-ZO (1:3)=CaOH-ZO (1:2)>CaOH-ZO (1:1)>CaOH-ZO (3:1)=CaOH-ZO (2:1)>CaOH=ZO-eugenol. Order of minimum bactericidal concentration, which shows a weaker bactericidal effect, according to type of paste, were as followed: CaOH-ZO (1:3)>CaOH-ZO mixture (1:2)>CaOH-ZO mixture (1:1)>CaOH-ZO mixture (3:1)=CaOH-ZO (2:1)>CaOH=ZO-eugenol. Only CaOH-ZO (1:3) and CaOH-ZO (1:2), showed significantly weaker MICs and MBCs (p < 0.001). Considering the limitations of an in-vitro study, in terms of anti-bacterial effects against Enterococcus faecalis, CaOH-ZO mixture (2:1) is equivalent to ZO-eugenol as the most commonly used material in polypectomy of primary teeth.

Staphylococcus aureus, as an opportunistic pathogen, is the cause of a variety of diseases from mild skin infections to severe invasive infections and food poisoning. Increasing antibiotic resistance in S. aureus isolates has become a major threat to public health. The use of compounds produced by probiotics can be a solution to this problem. Thus, the purpose of this study was to investigate the effect of Saccharomyces cerevisiae on some virulence factors (biofilm, α-hemolysin, and enterotoxin A) of S. aureus. Supernatant and lysate extracts were prepared from S. cerevisiae S3 culture. Sub-MIC concentrations of both extracts were separately applied to S. aureus ATCC 29213 (methicillin-sensitive S. aureus; MSSA) and S. aureus ATCC 33591 (methicillin-resistant S. aureus; MRSA) strains. Biofilm formation of these strains was measured by microtiter plate assay and expression level of α-hemolysin and enterotoxin A genes (hla and sea, respectively) using real-time PCR technique. The supernatant extract has reduced both biofilm formation and expression of sea and hla genes, while lysate extract had only anti-biofilm effects. The MRSA strain showed more susceptibility to yeast extracts than MSSA strain in all tests.


The present study exhibited favorable antagonistic effects of S. cerevisiae S3, as a probiotic yeast, on MSSA and MRSA strains. Based on the findings of this study, the compounds produced by this yeast can be used to control S. aureus infections; however, further similar studies should be conducted to confirm the findings of the present study.

Between 2007 and 2011, the mortality rate for burns patients at Dr. Soetomo General Hospital, Surabaya, Indonesia was 14.1% and 60% were suspected to be sepsis-related. Immunosuppression, gut barrier disruption, and intestinal hypomotility cause bacterial and bacterial product translocation. Probiotics improve the intestinal microbiome and eventually reduce bacterial translocation, and an increased secretory immunoglobulin A (SIgA) secretion post-administration of a multi-species probiotic has been observed. We aimed to determine whether a single-strain probiotic administration could show strengthened intestinal immunity, through an increase in SIgA levels, as with multi-strain probiotics. Sixteen burns patients from our hospital Burns Centre were randomized into three treatment groups, and the patients were administered either a placebo, a Lactobacillus reuteri protectis probiotic, or a Bifidobacterium infantis 35624 probiotic for 14 consecutive days. The SIgA levels were analyzed using ELISA pre- and post-treatment. The post-treatment SIgA levels in the placebo, Lactobacillus reuteri protectis probiotic, and Bifidobacterium infantis 35624 probiotic groups were 222.56±74.22 mg/dL, 223.92±68.89 mg/dL, and 332.38±64.27 mg/dL, respectively. Decreased SIgA levels were observed in the placebo (7.19±15.87) and in the Lactobacillus reuteri protectis probiotic (1.9920±14.76) groups, whereas an increase was seen in the SIgA level in the Bifidobacterium infantis 35624 probiotic group (58.26±77.41). The Bifidobacterium infantis 35624 single-strain probiotic is generally superior to Lactobacillus reuteri protectis in altering intestinal immunity; however, this finding was not statistically significant. A multi-strain probiotic supplement is recommended for burns patients.

The intestinal microflora has an important role in the health status. Since probiotics can balance the intestinal microflora, they have a lot of health beneficial effects. So the appropriate selection of probiotics can cause health-promoting effects. In this study, the combined effects of Bacillus subtilis and Bacillus coagulans on the intestinal microflora and growth performance in rats were investigated. 80 male rats were divided into the treatment (receiving 5×107 spores/ml of B. subtilis and 5×107 spores/ml of B. coagulans for three weeks in daily water) and control (tap water without probiotics) groups. The total aerobic and anaerobic microorganisms, lactic acid bacteria (LAB), coliforms and spores were weekly counted in the fecal samples. Additionally, the water and feed consumption, the weight gain and Feed Conversion Ratio (FCR) were calculated for each week. The probiotics significantly increased the total aerobic, LAB and spore counts and caused significant reduction in the anaerobe and coliform counts. Following three weeks of probiotic administration, the number of anaerobic bacteria, and coliforms were reduced by up to 0.7 and 1.18 log and the number of aerobic bacteria, LAB and spores were increased by 1.29, 1.15 and 7.2 log respectively. Also, the results showed the feed consumption reduction, weight gain and FCR enhancement in the probiotic group (p < 0.05). Supplementation the spores of B. subtilis and B. coagulans improved the growth performance and was beneficial to the intestinal microbiota in rats.