Diversity of halophilic and halotolerant bacteria in the largest seasonal hypersaline lake (Aran-Bidgol-Iran).

PURPOSE: In this study, the culturable halophilic and halotolerant bacterial diversity was determined in Aran-Bidgol as a thalassohaline seasonal hypersaline lake in Iran. METHODS: Thirty water, soil, sediments, coastal mud, multi-color brines and salt crystals samples were extracted and cultured using different media and incubation conditions. Totally 958 isolates were obtained and 87 isolates were selected for further studies, based on morphological, physiological and biochemical tests, representing different morphotypes. RESULTS: Based on 16S rRNA gene sequence analyses, the isolates exhibited 94.6-100% sequence similarity to the closest known species of the genera Bacillus, Halomonas, Oceanobacillus, Salinicoccus, Thalassobacillus, Ornithinibacillus, Halobacillus, Salicola, Virgibacillus, Aerococcus, Arthrobacter, Idiomarina, Paraliobacillus, Staphylococcus, Acinetobacter, Aneurinibacillus, Brevibacillus, Brevundimonas, Chromohalobacter, Gracilibacillus, Jeotgalicoccus, Kocuria, Marinilactibacillus, Marinobacter, Microbacterium, Paenibacillus, Paracoccus, Piscibacillus, Pseudomonas and Sediminibacillus and also, comparison of ARDRA patterns among the sequenced strains, using AluI, Bst UI and Hpa II enzymes showed that these patterns are in accordance with the phylogenetic position of these strains. CONCLUSION: The PCR-RFLP analyses suggested that ARDRA possess a functional potential for distinguishing halophilic bacteria to be used for further studies in elementary steps of isolation to reduce the tedious duplication of isolates.

Isolation and screening of extracellular anticancer enzymes from halophilic and halotolerant bacteria from different saline environments in Iran.

It was confirmed that several enzymes have anti-cancer activity. The enzymes L-asparaginase, L-glutaminase, and L-arginase were chosen according to amino acids starvation in cancer cells and screened in halophilic and halotolerant bacteria, given probably less immunological reactions of halophilic or halotolerant enzymes in patients. Out of 110 halophilic and halotolerant strains, isolated from different saline environments in Iran and screened, some could produce a variety of anticancer enzymes. A total of 29, 4, and 2 strains produced L-asparaginase, L-glutaminase, and L-arginase, respectively. According to the phenotypic characteristics and partial 16S rRNA gene sequence analysis, the positive strains-strains with the ability to produce these anticancer enzymes-were identified as the members of the genera: Bacillus, Dietzia, Halobacillus, Rhodococcus, Paenibacillus and Planococcus as Gram-positive bacteria and Pseudomonas, Marinobacter, Halomonas, Idiomarina, Vibrio and Stappia as Gram-negative bacteria. The production of anticancer enzymes was mostly observed in the rod-shaped Gram-negative isolates, particularly in the members of the genera Halomonas and Marinobacter. Most of the enzymes were produced in the stationary phase of growth and the maximum enzyme activity was experienced in strain GBPx3 (Vibrio sp.) for L-asparaginase at 1.0 IU/ml, strain R2S25 (Rhodococcus sp.) for L-glutaminase at 0.6 IU/ml and strain GAAy3 (Planococcus sp.) for L-arginase at 3.1 IU/ml. The optimum temperature and pH for L-asparaginase and L-glutaminase activities in selected strains were similar to the physiological conditions of human body and the enzymes could tolerate NaCl up to 7.5% concentration.

The Healing Effect of Sesame Oil, Camphor and Honey on Second Degree Burn Wounds in Rat.

BACKGROUND: Many studies were carried out to improve sophisticated dressings to accelerate healing processes and reduce the microbial burden in burn wounds. This study evaluated the healing effect of herbal ointment containing extract of sesame oil, camphor and honey on second degree burn wounds in rats in comparison with daily dressing oil vaseline. METHODS: Forty rats were randomly assigned to two equal groups. A deep second degree burn was formed on the back of each rat with using a standard burning technique. The burns were dressed daily with herbal ointment containing extract of sesame oil, camphor and honey in group 1, dressing oil vaseline in group 2. The response to treatment was evaluated by digital photography during the treatment on 0, 7, 14, 21, 28 days. Histological scoring was undertaken for scar tissue samples on 0, 7, 14, 21, 28 days. RESULTS: Considerable epithelization in the herbal ointment group vs. the control group over the study period was noted. Neovascularization was significantly higher in herbal ointment treated rats as well. In terms of difference of wound surface area, maximal healing was noticed in herbal ointment extract of sesame oil, camphor and honey group and the minimal repair in the control group. CONCLUSION: The greatest rate of healing was in the herbal ointment group containing sesame oil, camphor and honey, so the herbal ointment as a suitable substitute for dressing and healing of burn wound injuries is recommended.

Erratum: High carotenoid production by a halotolerant bacterium, Kocuria sp. strain QWT-12 and anticancer activity of its carotenoid.

[This corrects the article on p. 840 in vol. 16, PMID: 28827999.].

High carotenoid production by a halotolerant bacterium, Kocuria sp. strain QWT-12 and anticancer activity of its carotenoid.

Halophilic prokaryotes are extremophile microorganisms that grow optimally in media containing salts and almost appeared pigmented. Many of them contain high concentrations of carotenoids. Amongst 15 strains of halophilic prokaryotes isolated from industrial tannery wastewater in Qom, a Gram-stain-positive coccoid, aerobic, non-endospore-forming, halotolerant bacterium designated as strain QWT-12 showed high capacity in the production of carotenoids in a wide range of culture medium factors. 16S rRNA gene analysis showed that this strain belongs to the genus Kocuria. Carotenoids from this strain were extracted by methanol. MTT assay for extracted carotenoid was carried out against seven cancer cell lines belonging to breast, lung and prostate cancer with negative control of fibroblast cell line through six concentration levels to find out IC50. Based on statistical analysis of data from MTT assay, IC50 of 1, 4 and 8 mg/ml for MCF-7, (A549 and MDA-MB-468) and MDA-MB-231 respectively. Additionally, qualitative carotenoid determination was carried out using spectrophotometric method in 300-600 nm and thin layer chromatography, respectively. According to the obtained results from mass spectrophotometry, absorption spectrum of strain QWT-12 is similar to the absorption spectrum of the carotenoid neurosporene.

Analysis of Healing Effect of Alginate Sulfate Hydrogel Dressing Containing Antimicrobial Peptide on Wound Infection Caused by Methicillin-Resistant Staphylococcus aureus.

BACKGROUND: Wound infections caused by methicillin-resistant Staphylococcus aureus are a health problem worldwide; therefore, it is necessary to develop new antimicrobial compounds. Considering broad-spectrum antimicrobial activity and low probability of drug resistance to peptides, applications these peptides are being studied extensively. OBJECTIVES: In this study, to control drug release over time, an alginate sulfate-based hydrogel impregnated with the CM11 peptide as the antimicrobial agent was developed, and its healing effects were tested on skin infections caused by methicillin-resistant S. aureus strains in a mouse model. MATERIALS AND METHODS: Minimum inhibitory and minimum bactericidal concentrations of the CM11 peptide and alginate hydrogel in combination with the peptide were determined. Forty mice were divided into 4 groups: 1 group as a negative control (without treatment; however, 5 mice received hydrogel dressing without peptide), 1 group as a positive control (2% mupirocin treatment), and 2 groups as test groups. To establish skin infection, 200 µL of bacterial suspension with 3 × 10(8) CFU/mL concentration was subcutaneously injected in the scapular region of the mice. On the basis of the in vitro minimal bactericidal concentration of the alginate hydrogel containing peptide for 15 clinical isolates, hydrogel containing 128 mg/L of peptide was used for wound dressing over an 8-day period. RESULTS: The highest and lowest numbers of wounds were observed on day 2 in the negative and positive control groups, respectively. During the 8-day period, the positive control and hydrogel containing peptide treatment groups showed similar levels of wound healing. CONCLUSIONS: This study showed that compared to standard drug treatment, treatment with hydrogel containing peptide had substantial antibacterial effects on S. aureus wound infections in mice.

Comparison of bacterial biodiversity and enzyme production in three hypersaline lakes; urmia, howz-soltan and aran-bidgol.

This research is a comparative study on the diversity of halophilic bacteria with hydrolytic activities in three significant hypersaline lakes; Urmia in the northwest and Howz-Soltan and Aran-Bidgol in the central desert in Iran. Isolated strains from these saline lakes were found to be halotolerant, moderately and extremely halophilic bacteria. The bacteria in each saline lake were able to produce different hydrolytic enzymes including amylase, protease, lipase, DNase, inulinase, xylanase, carboxy methyl cellulase, pectinase and pullulanase. 188, 302, 91 halophilic strains were isolated from Urmia Lake, Howz-Soltan and Aran-Bidgol playa, respectively. The numbers of Gram-positive strains were more than Gram-negatives, and among Gram-positive bacteria; spore-forming bacilli were most abundant. Due to the unique physico-chemical conditions of the lake environments, the hydrolytic activities of isolated strains were significantly different. For instance, isolated strains from Howz-Soltan playa did not produce pectinase, DNase, amylase, lipase and inulinase, while the isolates from Aran-Bidgol playa had a great ability to produce pectinase and DNase. The strains from Urmia Lake were also good producers of DNase but failed to show any chitinase activity. The diversity of halophilic bacteria from the mentioned three saline lakes was also determined using PCR-amplified 16S rRNA followed by phylogenetic analysis of the partial 16S rRNA sequences.