Iranian Journal of Microbiology
Volume:3 Issue: 3, Sep 2011

Salmonella is one of the leading causes of food-borne diseases. Increasing occurrence of antimicrobial resistance, especially multidrug-resistance, in Salmonella serovars is a major public health problem worldwide. This study was carried out to detect class I integrons and antibiotic resistance profiles in clinical isolates of Salmonella serovars collected from seven hospitals in Tehran during November 2009 to June 2010. Antibiotic susceptibility profile of 19 antibiotics against 58 Salmonella isolates commonly used in humans was determined using disk diffusion assay. Minimum inhibitory concentration against ceftriaxone and ciprofloxacin was studied. PCR assays were used to detect class I integrons. Among 58 Salmonella isolates, 72.4% were Salmonella enterica serovar Enteritidis, 8.7% were Salmonella enterica serovar Typhimurium and 18.9% were other serovars. Of the total 58 Salmonella serovars, 43 (74.1%) were multidrug-resistant and showed resistance to three or more antibiotic families. Class I integrons were identified in 38 (88.3%) MDR Salmonella isolates. Ciprofloxacin minimum inhibitory concentration ranged between 0.125-2 μg⁄ml for four isolates and other four isolates exhibited resistance to ceftriaxone (MIC 64-256 μg ⁄ml). The high prevalence of class I integrons was seen in our MDR Salmonella isolates and class I integrons might play an important role in the dissemination of antimicrobial resistance determinants.

Pyrococcus woesei is a hyperthermophilic archaea and produces a heat stable polymerase (Pwo polymerase) that has proofreading activity. In this study, this microorganism was cultured, its DNA was extracted and the pwo gene polymerase was cloned, expressed and purified. The DNA sequence of the cloned gene was verified by sequencing. The pwo polymerase gene consists of 2,328 bps (775 amino acids with about 90 kD molecular weight). Cloning was done by GATEWAYTM Cloning System and for purification of recombinant protein; His6x-Tag was added to the C-terminus of the recombinant protein.Results and We could purify Pwo polymerase enzyme by Ni-NTA resin. PCR assay showed that Pwo polymerase activity is comparable to a commercial Pfu polymerase activity.

As prenatal screening for sexually transmitted infections and treatment of infected pregnant women is not routinely performed in Iran and prevalence of two sexually transmitted pathogens, Chlamydia trachomatis and Mycoplasma genitalium, in Sabzevar (east of Iran) is unknown, we decided to perform this prospective study. One hundred ninety-six urine specimens of pregnant women attending the specialized maternity hospital of the city were collected and tested by duplex PCR. A total of 31 specimens were positive (15.81%) (27 Chlamydia trachomatis isolates, 13.77%; and 2 Mycoplasma genitalium isolates, 1.02%). Co-infection with both species was detected in 2 specimens (1.02%). A significant correlation was found between preterm labor and infection (P-value ≤ 0.05). The present study shows high prevalence of Chlamydial infections in comparison with Mycoplasma genitalium in this region. Further studies with larger sample size and more focused on different groups at risk are needed for a movement towards prevention and control of sexually transmitted infections (STIs).

Probiotics are live microbial feed supplements which beneficially affect the host animal by improving its intestinal microbial balance, producing metabolites which inhibit the colonization or growth of other microorganisms or by competing with them for resources such as nutrients or space. The aim of this study was to investigate the probiotic properties of Candida famata and Geobacillus thermoleovorans. In this study, yeast and bacterial strains isolated from pure oil waste were identified using Api 50 CHB and Api Candida Systems and their probiotic properties were studied through antimicrobial activity, biofilm production, adherence assay and enzymatic characterization.Results and According to biochemical analyses, these strains corresponded to Geobacillus thermoleovorans and Candida famata. Antagonism assay results showed that the tested strains have an inhibitory effect against tested pathogenic bacteria. The yeast Candida famata was unable to produce biofilm on Congo Red Agar (CRA), while the bacterial strain was a slime producer. Adherence assays to abiotic surfaces revealed that the investigated strains were fairly adhesive to polystyrene with values ranging from 0.18 to 0.34 at 595 nm. The enzymatic characterization revealed that the tested strains expressed enzymes such as phosphatase alkaline, esterase lipase (C8), amylase, lipase, lecitenase and caseinase. The obtained results may allow the isolated strains to be considered as having the potential to be candidate probiotics.

Leptospirosis is an important zoonotic disease caused by Leptospira interrogans. Leptospirosis leads to economical losses in dairy farm industry. The objective of this study was to evaluate the pathogenic serovars of Leptospira interrogans in dairy cattle herds of Shahrekord by PCR. Two hundred samples (100 urine and 100 blood) were collected from 100 cows randomly and delivered to the laboratory. Samples were stored at -20 °C. DNA was extracted and purified from the plasma and urine samples and concentrated on diatoms in the presence of guanidine thiocyanate (GuSCN). PCR products were detected and identified as Leptospira by ilumination of the expected size of DNA bands after staining of the agarose gel with ethidium bromide gels. PCR products were purified and sequenced. The results showed that 28% of urine samples and 23% of plasma samples were contaminated. The major serotypes were Icterohaemorrhagiae (50%) and Pomona (37.5%). The urine samples of 17 cows were positive for Leptospira without positive plasma samples. This indicated that these cows are reservoirs in dairy herds of Shahrekord and dangerous for human health. The plasma samples of twelve cows were positive for Leptospira without positive urine samples. Leptospira serotypes can be maintained in relatively dry regions and must be considered when dealing with leptospirosis in dairy farms of Shahrekord and human health.

To find antagonistic bacteria with potential antifungal activity against some pathogenic fungi, including Aspergillus niger, A. flavus, Fusarium moniliforme and Penicillium marneffei, a total of 148 agricultural soil samples from different sites of Tehran were examined. Antagonistic soils were selected by screening against A. niger on glucose-yeast extract (GY) agar using a visual agar plate assay method. All growing bacteria were examined for antifungal activity, and antagonistic bacteria identified based on 16S rRNA sequence analysis. Among a total number of 97 bacteria isolated form inhibitory soils (36 samples), 16 bacteria were reported as strong growth inhibitors in co-cultures on GY agar with all tested fungi at variable degrees. Fungal growth inhibitory bacteria were cultured against all fungi and growth inhibition was measured and analyzed between test and control groups by statistical analysis (ANOVA). Molecular identification of antagonistic bacteria indicated that most bacterial isolates belonged to the genus Bacillus (81.25%), including B. subtilis (5 isolates), B. amyloliquefaciens (6 isolates) and B. valismortis (2 isolates), followed by one isolate (6.25%) from each Streptomyces sp., Pseudomonas chlororaphis and Acinetobacter baumannii. Based on the visual plate assay results, total fungal growth inhibition of all bacteria was reported in the range of 13.2 to 68.3%. P. chlororaphis S105 was reported as the most potent antagonistic bacterium which inhibited the growth of A. niger by 68.3%, followed by F. moniliforme (66.4%), A. flavus (64.7%) and P. marneffei (57.1%). P. chlororaphis and some other inhibitory bacteria reported in the present study, they may be considered not only as a rich source of useful metabolites with potential application in antifungal drug discovery, but also as potential candidates for biological control programs.

Aflatoxins are mycotoxins produced by Aspergillus flavus and Aspergillus parasiticus that can contaminate human and animal foods, including corn, wheat, rice, peanuts, and many other crops resulting in the illness or death of human and animal consumers. The aim of this study was to detect aflatoxin B1, B2, G1, G2 and total aflatoxin in Kashkineh, a traditional Iranian food. This survey was conducted to detect aflatoxins on 41 samples of Kashkineh. The samples were randomly collected from traditional bazaars and supermarkets of Khorramabad city of Iran. The presence and quantity of aflatoxins was determined by high performance liquid chromatography (HPLC). The average concentrations of AFB1, AFB2, AFG1, and AFG2 in all samples and in a mixed sample of all samples were not detectable (ND). The only sample that showed aflatoxin contamination was sample number 29 of which the AFB1 concentration was 0.64 ng/g. Although some people believe Kashkineh is carcinogenic due to toxins, this study showed kashkineh is not contaminated with aflatoxins.

Pulsed electric field (PEF) is a novel emerging technology which is believed to have the potential to substitute conventional thermal pasteurization (HTST). In the current study PEF was compared with HTST based on microbial inactivation and quality attributes. Juice was prepared by extracting it from Semirum apples. They were chilled to 4ºC over night. Then were divided into two lots, one was treated by PEF and the other by HTST. The treated juices were cultured on tryphtic soy broth (TSB) and results were recorded for 168 days. Quality changes were characterized by color and sensory test. Color changes were quantified using Hunter Lab equipment and equation. Sensory changes were evaluated by test panelists. Using selective media E. Coli was enumerated, the total count of the organism was noticeably lower than PEF treated specimen and after 168. The count didnt reach the initial population. Whereas in PEF treated juice bacterial count bounced back to the initial count and exceeds. Results from Hunter Lab indicated a of 3.04 and 3.08 system for PEF and HTST treated juices. Sensory panel showed that PEF is superior to thermal treatment. The study indicated HTST is more suitable based on food safety encounters. However PEF treated are closer to fresh juices based on quality factors. It can be concluded that PEF has the potential to become a suitable replacement to conventional process if improvements in design are applied.

An immunosuppressed man was admitted to hospital with diarrhea and a history of urinary tract infection. He was subjected to treatment with antibiotics. The patient died of putative severe sepsis. The etiological agent was a carbapenemase producing isolate of Bacillus circulans with resistance to all prescribed antimicrobial agents.