Iranian Journal of Microbiology
Volume:4 Issue: 1, Mar 2012

In December 2010 four, lions and one tiger died at the Tehran zoo. Out of all samples, Burkholderia mallei (causative agent of Glanders) was isolated just from ulcer sample of the tiger which was imported to Iran from Russia. One nasal swab from a tiger and fifteen blood samples with anticoagulant belonging to one tiger and fourteen lions (four dead lions and eleven live lions) were collected and were inoculated directly onto the selective media. The isolate was identified by morphological and biochemical and API BBL tests and PCR using specific primers (Bma-IS407-flip). The standard (Razi Type Culture Collection RTCC: 2375) and tiger isolates were inoculated into 2 guinea pigs. All residue solipeds and carnivores were checked by Malleination test and Complement Fixation (CF) Test respectively. One isolate of B. mallei was isolated from tiger’s nasal swab. Both of B.mallei strains were recovered from inoculated animals. All of solipeds were negative by malleination test, however, 11 lions including 4 dead and 7 live lions out of 14 lions were positive in CF test for Glanders and all were put down by the authorities. Active surveillance of Glanders is essential for solipeds, especially it’s more important while being used to feed valuable carnivores like lions and tigers. Therefore, a reliable test like malleination must be carried out twice (first before transferring and one month after quarantine). Both test results should be negative for use for feeding.

HIV-1 and HCV infections are life threatening problems in patients who receive blood products. Serological methods have proven useful in detecting these infections, but there are setbacks that make it challenging to detect these infectious agents. By the advent of Nucleic Acid Testing (NAT) methods, especially in multiplex format, more precise detection is possible. We have developed a multiplex RT-PCR assay for simultaneous detection of HIV-1 and HCV. Primers were designed for highly conserved region of genome of each virus. Using these primers and standard plasmids, we determined the limit of detection, clinical and analytical specificity and sensitivity of the assay. Monoplex and multiplex RT-PCR were performed. Analytical sensitivity was considered to be 100 and 200 copies/ml for HIV-1 and HCV, respectively. High concentration of one virus had no significant effect on the detection of the other one with low concentration. By analysis of 40 samples, clinical sensitivity of the assay was determined to be 97.5%. Using different viral and human genome samples, the specificity of the assay was evaluated to be 100%. The aim of this study was to develop a reliable, rapid and cost effective method to detect HIV-1 and HCV simultaneously. Results showed that this simple and rapid method is perfectly capable of detecting two viruses in clinical samples.

The significant rise in food borne infections is mainly caused by Campylobacter spp., Salmonella serovars and Verotoxigenic Escherichia coli. As the emerging food borne pathogens cause disease, more studies have been conducted for rapid detection of these pathogens. The combination of immunomagnetic separation and polymerase chain reaction (IMS-PCR) is the most accurate and rapid test preferred by almost every researcher. Fourier Transform Infrared Spectroscopy (FTIR) is preferred for being a new, user friendly and rapid technique in microbiological analyses. The main aim of this study is to detect application of IMS-FTIR for Salmonella identification from foods in a short time with a higher sensitivity. Conventional Culture Technique (CC), IMS-CC, IMS-PCR and IMS-FTIR techniques were compared with each other for rapid detection in artificially contaminated minced beef with Salmonella Typhimurium, as of the 2nd, 4th and 8th hours of contamination. The method was evaluated in different food matrices and sensitivity, specifity and overall recovery was calculated. The results indicate that IMS-FTIR can detect S. Typhimurium as of the 8th hour with sensitivity of 95.6667, accuracy of 91.69329, false positive ratio of 0.04333 and overall recovery of 95.66%. It can be suggested that the IMS-FTIR method is capable of detecting S.Typhimurium in a short time with lower cost.

The objective of this study was to monitor the microorganisms isolated from grapes and its derivative traditional products produced in Iran. Four kinds of grapes cultivated summer of 2010 in vineyard of Takestan and also grape derived products from Shahrod, Hamedan and Takestan were used for this study. The samples were cultured in specific media to isolate the microorganisms that might grow on or pollute the products. Species of bacteria and fungi isolated from 4 kinds of grapes cultivated in Takestan graveyards and also from 2 kinds of derived traditional products; grape sap and sour grape (abe-ghure locally named), were taken from Takestan, Shahrod and Hamedan cities. Also, bacteria Bacillus spp., Micrococcus spp., Clostridium spp., and fungus of Penicillium spp., and Aspergillus spp. were isolated. The isolated bacteria were common microorganisms that grow in soil or in the organic fertilizer and may appear from the environments that samples were collected. These bacteria were not pathogenic to human. The fungus isolated from the grapes may harm humans as they produce toxin. The results suggested that bacterial diversity on grapes and its derived traditional products are expected to be monitored and described in all Iranian graveyards as Iran has been known as one of the world’s biggest grape producers.

In this study, the antibacterial effect of essential oil of tarragon (Artemisia dracunculus) on Staphylococcus aureus and Escherichia coli was evaluated in culture media and Iranian white cheese. The tarragon essential oil (EO) obtained by the steam distillation method and its antibacterial activity was evaluated in 96-well microtiter plates containing brain heart infusion broth. The enumeration of S. aureus and E. coli in cheese samples were carried out on the following media: Baired parker agar for S.aureus, incubated at 37 °C for 24 h; and MacConkey sorbitol agar for E. coli, incubated at 37°C for 24 h. Iranian white cheese was produced from fresh and whole pasteurized cow milk (2.5%). Bacteria (103 cfu/mL) were inoculated to different batches. Cheese was treated with different concentrations of EO (15 and 1500 μg/mL) and separated into four parts in an equal manner. The sensory evaluation was done by a panel of four judges. According to the results obtained, minimum inhibitory concentrations (MIC) for E. coli and S. aureus were 2500 and 1250 μg/mL, respectively. Also, minimum bactericidal concentration (MBC) for the mentioned microorganisms were 5000 and 2500 μg/mL, respectively. All the EO concentrations for each bacteria result in reducing bacterial count of cheese samples compared to control (P < 0.05). Also, with increasing concentration of EO in cheese samples, the bacterial count was reduced further (P < 0.05). Based on our findings, tarragon essential oil has antibacterial effect on two important pathogen bacteria (S. aureus and E. coli) and can be applied as a preservative in foods such as cheese.

Cryptosporidiosis has not been reported as an endemic disease in Kashmir, but high prevalence of Cryptosporidium sp. has been found among asymptomatic (non-diarrheic) HIV positive immigrants in present study. Due to increasing number of HIV positive immigrants in Kashmir, Cryptosporidium may become a public health problem in Kashmir. A total of 45 stool samples were obtained from symptomatic (diarrheic n = 9) and asymptomatic (non-diarrheic n = 36) patients infected with HIV. The stool samples were concentrated using formalin ethyl acetate concentration technique, stained with modified Kinyoun’s cold stain and oocysts were identified by microscopy under 1000 x magnification. It was confirmed by detection of antigens in stool samples by ELISA. It was established that all the patients studied were carriers of Cryptosporidium. In present study though 80% of patients were asymptomatic (non-diarrheic) and HIV positive which involved non-Kashmiri army personals and travelers (immigrants) but were carriers of Cryptosporidium and 20% of HIV positive patients were emigrants (local Kashmiri traders) who travelled different states of India were having diarrhea (symptomatic) as well as carrier of Cryptosporidium. Though Cryptosporidium infection causes chronic diarrhea but in present study all HIV positive patients screened whether diarrheic or non-diarrheic were positive for Cryptosporidium. To prevent the transmission of Cryptosporidium oocyst in environment and endemic spread of cryptosporidiosis as non-diarrheic HIV positive population may be potential source of infection, obligatory laboratory testing for Cryptosporidium in HIV positive immigrant population like traders and travelers is highly recommended in order to have a better understanding of the cause of spread Cryptosporidium infection in Kashmir.

A chronic fungal infection in tropical regions, chromoblastomycosis is caused by dematiaceous fungi, the form-family of Fungi Imperfecti, usually affecting one lower limb at the site of a trauma but sometimes involving other areas of the body including head & neck. In this article, we report the case of a rare primary chromoblastomycosis of the palate and chest in a 27-year-old man who was successfully treated with surgical resection and combined drug therapy, and eventually free tissue transfer reconstructive surgical procedure to cure the palatine defect.

A modified pulsed-field gel electrophoresis (PFGE) protocol was developed and applied to clinical isolates of Mycobacterium tuberculosis complex to reduce the cost of using lyticase. This protocol reduces the expense of PFGE typing of Mycobacterium tuberculosis complex as it removes the use of lyticase during the spheroplast formation from these bacteria.