Evaluation of Thirty Microalgal Isolates as Biodiesel Feedstocks Based on Lipid Productivity and Triacylglycerol (TAG) Content.

Microalgae are considered feedstock for biodiesel production due to their capability to accumulate triacylglycerols, which have a 99% conversion rate into biodiesel, under certain conditions. This study aims to evaluate thirty native microalgal strains as feedstock for biodiesel production based on their biomass and lipid productivities, and total lipid and triacylglycerol contents under nitrogen-sufficient and nitrogen starvation conditions. In addition, Chlamydomonas reinhardtii cw15 mutant strain was utilized as a reference strain for triacylglycerol accumulation. Among the eight potent strains, Chlorella vulgaris KP2 was considered as a most promising strain with the highest triacylglycerol content, highest total lipid content (28.56% of dry cell weight), and the highest lipid productivity (4.56 mg/L/day) under nitrogen starvation. Under nitrogen starvation, the major fatty acids in the triacylglycerol of Chlorella vulgaris KP2 were C18:1 (37.56%), C16:0 (23.16%), C18:0 (23.07), C18:2 (7.00%), and C18:3 (3.12%), and the percentages of saturated fatty acids, monounsaturated fatty acids, and polyunsaturated fatty acids represented 49.26, 38.73, and 10.12% of the total fatty acids, respectively. Furthermore, the fatty acid methyl esters of triacylglycerol displayed remarkable biodiesel properties with a lower iodine value (59.00 gI2/100 g), higher oxidative stability (14.24 h) and higher cetane number (58.73) under nitrogen starvation. This study suggests that nitrogen-starved Chlorella vulgaris KP2 could be used as a feedstock for biodiesel production due to the considerable amounts of triacylglycerol and favorable biodiesel properties.

Determination of the acute toxic effects of zinc oxide nanoparticles (ZnO NPs) in total hemocytes counts of Galleria mellonella (Lepidoptera: Pyralidae) with two different methods.

Zinc oxide nanoparticles (ZnO NPs) are now commonly used in many consumer products (detergents, antibacterial products, protective creams). The aim of the study is to determine the ecotoxicological effects of ZnO NPs on the survival and in the total hemocyte counts of Galleria mellonella L. 1758 (Lepidoptera: Pyralidae) with two different methods (automated cell counter and hemocytometer). A toxicity test was performed to determine the lethal concentrations of ZnO NPs on larvae by force feeding method. After 24 h the treatment, LC50 was 6.03 µg/10 µl and LC99 was 12.86 µg/10 µl for force fed larvae according to probit assay. The NP that induced changes in the total hemocytes counts were counted by optical microscopy (larvae exposed to four different doses of ZnO NPs) and by automated cell counter. Analyses of total hemocyte counts of the insect were performed with four doses (0.5, 1, 2.5, 5 µg/10 µl)

The evaluation of Starmerella magnoliae X3 as a biodiesel feedstock based on triacylglycerol (TAG) production, lipid productivity, and fatty acid profile under nitrogen limitation and acidic pH conditions.

The effects of four initial culture pH values (3, 4, 5, and 6) and nitrogen limitation on growth, TAG accumulation, lipid production, fatty acid profile, and estimated biodiesel quality of Starmerella magnoliae X3 were investigated. TAG and lipid levels were measured by Nile Red fluorescence and sulfo-phospho-vanilin (SPV) techniques, respectively. The results showed that a combination of nitrogen limitation and acidic pH significantly (p < 0.05) increased TAG accumulation, total lipid contents, and lipid productivity in Starmerella magnoliae X3 compared to the control group. Under nitrogen limitation, the highest TAG accumulation was achieved at initial pHs of 3 and 5 after 72 h of cultivation, and the highest lipid productivity (0.306 g L-1 d-1) was observed after 48 h at pH 3; the major fatty acids at the four pH values were oleic acid (63.6%-64%), palmitoleic acid (11.3%-12.5%), stearic acid (9.7%-11.4%), and palmitic acid (9.4%-10%). In addition, both stresses were associated with lower iodine value and higher cetane number of the biodiesel compared to the control. These findings suggest that cultivation in a low-nitrogen medium at an initial pH of 3 or 5 holds promise in increasing TAG production in Starmerella magnoliae X3.

Toxicity of herbicides to cyanobacterial isolates.

The herbicides most commonly used in Turkey are trifluralin (alpha,alpha,alpha-trifluoro-2,6-dinitro-N,N-dipropyl-p-toluidine), 2,4-D (2,4-dichlorophenoxyacetic acid), and linuron [3-(3,4-dichlorophenyl)-N-methoxy-N-methylurea]. The effects of these three herbicides on the growth of 10 threatened aquatic cyanobacterial isolates were tested by using 9-day exposure experiments at concentrations of 50-200 mg l(-1) for trifluralin and 2,4-D and 0.05-1 mg l(-1) for linuron. Concentrations of herbicides that elicited a 50% growth reduction over 9 days (EC50) were 136-882 mg l(-1) trifluralin, 122-747 mg l(-1) 2,4-D, and 0 002-0.714 mg l(-1) linuron. Synechocystis sp. H6 was more tolerant to the three herbicides than the other isolates of cyanobacteria. Chroococcus sp. S27, Microcystis sp. S17, and Synechococcus sp. S24 were the most sensitive to trifluralin, 2,4-D, and linuron, respectively. There has been increasing awareness about using cyanobacteria as pollution control agents. The present study indicated that as the concentrations of the these herbicides were increased, significant changes were recorded in cyanobacterial growth rates. Results obtained from this comparative study allow the choice of suitable herbicides for agricultural practices based on their effects on cyanobacterial growth.

Relationship between chromium(VI) resistance and extracellular polymeric substances (EPS) concentration by some cyanobacterial isolates.

BACKGROUND, AIM, AND SCOPE: Chromium(VI) resistance and its association with extracellular polymeric substance (EPS) concentration in cyanobacteria was investigated. Increased EPS concentration was associated with Cr(VI) resistance. The most resistant isolate, Chroococcus sp. H(4), secreted the most EPS (427 mg/L). MATERIALS AND METHODS: EPS concentration of the two most resistant isolates (Chroococcus sp. H(4) and Synechocystis sp. S(63)) was investigated following exposure to 15 and 35 ppm Cr(VI). The composition of EPS produced by Chroococcus sp. H(4) following exposure to 10 ppm Cr(VI) was analyzed using high-performance liquid chromatography. Control EPS was composed of glucose (99%) and galactronic acid (1%); in the presence of 10 ppm Cr(VI), EPS composition changed to glucose (9%), xylose (75%), rhamnose (14%), and galacturonic acid (2%). RESULTS AND DISCUSSION: Results indicated that (1) exposure to elevated concentrations of Cr(VI) affected the composition of EPS produced by Chroococcus sp. H(4), and (2) there was a correlation between Cr(VI) resistance and EPS concentration in some cyanobacteria.

Antiproliferative, neurotoxic, genotoxic and mutagenic effects of toxic cyanobacterial extracts.

Cyanobacteria are the rich resource of various secondary metabolites including toxins with broad pharmaceutical significance. The aim of this work was to evaluate the antiproliferative, neurotoxic, genotoxic and mutagenic effects of cyanobacterial extracts containing Microcystin-LR (MCLR) in vitro. ELISA analysis results showed that MCLR contents of five cyanobacterial extracts were 2.07 ng/mL, 1.43 ng/mL, 1.41 ng/mL, 1.27 ng/mL, and 1.12 ng/mL for Leptolyngbya sp. SB1, Phormidium sp. SB4, Oscillatoria earlei SB5, Phormidium sp. SB2, Uncultured cyanobacterium, respectively. Phormidium sp. SB4 and Phormidium sp. SB2 extracts had the lowest neurotoxicity (86% and 79% cell viability, respectively) and Oscillatoria earlei SB5 extracts had the highest neurotoxicity (47% cell viability) on PC12 cell at 1000 µg/ml extract concentration. Leptolyngbya sp. SB1 and Phormidium sp. SB2 showed the highest antiproliferative effect (92% and 77% cell death) on HT29 cell. On the other hand, all concentrations of five toxic cyanobacterial extracts induced DNA damage between 3.0% and 1.3% of tail intensity and did not cause any direct mutagenic effect at the 1000 µg/plate cyanobacterial extracts. These results suggest that cyanobacteria-derived MCLR is a promising candidate for development of effective agents against colon cancer.

Metal removal of cyanobacterial exopolysaccharides by uronic acid content and monosaccharide composition.

In the present study, chromium, cadmium and metal mixed (chromium+cadmium) removal and its association with exopolysaccharides and uronic acids production in Synechocystis sp. BASO671 were investigated. It was investigated that BASO671 showed different removal ability when exposed to each metal solely and mixed metal. EPS production by BASO671 was increased following exposure to 15 and 35 ppm Cr(VI), Cd(II) and Cr(VI)+Cd(II). Monomer composition of EPS was changed after metal treatment. Uronic acid contents of metal treated cells were higher than control cells of each isolate. Also, glucuronic acid content and galactronic acid content of EPS correlated with uronic acid contents of cells. Scanning electron microscopy and energy dispersive X-ray spectroscopy analysis confirmed that a considerable amount of metals had precipitated on the cell surface. Fourier transform infrared spectrum analysis of EPSs indicated the presence of CH and CO group, which may serve as binding sites for divalent cations.

Phenylalanine ammonia lyase (PAL) enzyme activity and antioxidant properties of some cyanobacteria isolates.

In the present study, six cyanobacteria isolates were evaluated for the PAL enzyme activity, and their methanol extracts were assessed for the total phenolic amount and other antioxidant parameters. Synechocystis sp. BASO444 and Synechocystis sp. BASO673 isolates with high levels of total phenols (66.0±1.2µg/mg, 78.1±1.8µg/mg, respectively) also showed high levels of PAL activities (20.5±3.1U/mg protein, 17.2±2.3U/mg protein, respectively) and strong antioxidant activities. To understand the effect of l-phenylalanine (l-phe) on the PAL activity, total phenolic amount, and phenolic constituents, isolates were evaluated with 100mg/l l-phe. While PAL activities exhibited no significant change with l-phe addition, total phenolic amount of the isolates significantly increased. HPLC analysis revealed gallic acid, trans-cinnamic acid, p-coumaric acid, and ferulic acid as the main compounds. Results suggested that the two isolate mights be an important source for the l-phe inducible phenolic compounds.

Removal and reduction of chromium by Pseudomonas spp. and their correlation to rhamnolipid production.

Chromium removal and its association with rhamnolipid production in Pseudomonas spp. were investigated. Three Pseudomonas spp. isolates (P. aeruginosa 78, P. aeruginosa 99, and P. stutzeri T3) were investigated with regard to their exposure to 10mg/L for chromium removal. P. aeruginosa 99 removed 16% and 20% more chromium than P. stutzeri T3 and P. aeruginosa 78 respectively. The reduction of Cr(VI) to Cr(III) by all the three isolates is more or less similar. P. aeruginosa 99, which removed higher chromium, also produced higher rhamnolipid (165±5 mg/mL). P. aeruginosa 78, which removed lower chromium, also produced lower rhamnolipid (126±3 mg/mL). Rhamnolipid production by P. aeruginosa 78 and P. aeruginosa 99 was increased in its exposure to 10mg/L chromium. In the present study, results showed that rhamnolipid might play a role in chromium removal by three Pseudomonas spp. isolates.

Phenolic composition and antimicrobial and antioxidant activities of Leucoagaricus leucothites (Vittad.) Wasser.

Leucoagaricus leucothites (Vittad.) Wasser is an edible macrofungus found limitedly in Turkey. The ethanolic extract of L. leucothites has shown strong antimicrobial activity against some foodborne and spoilage bacteria. Therefore, the phenolic composition of L. leucothites ethanolic extract was analyzed by using high-performance liquid chromatography. Catechin (288 mg/L) was found to be the major component in the extract. Antioxidant activity of the extract was evaluated by studying the scavenging ability of 2,2-diphenyl-1-picrylhydrazyl (DPPH) radicals, and chelating ability of the extract was assessed in a ferrous ion assay. The scavenging effect on DPPH radicals was 71% at 10 mg/mL, whereas the chelating effect on ferrous ions was 82% at 15 mg/mL. The total contents of phenol (3.40 ± 0.5 mg/g), ascorbic acid (2.10 ± 0.3 mg/g), ß-carotene (0.02 ± 0.0 mg/g), and lycopene (0.01 ± 0.0 mg/g) in the macrofungi ethanolic extract were also determined.

Modification of exopolysaccharide composition and production by three cyanobacterial isolates under salt stress.

BACKGROUND, AIM, AND SCOPE: Polysaccharides are renewable resources representing an important class of polymeric materials of biotechnological interest, offering a wide variety of potentially useful products to mankind. Exopolysaccharides (EPSs) of microbial origin with a novel functionality, reproducible physico-chemical properties, stable cost and supply, became a better alternative to polysaccharides of algal origin. EPSs are believed to protect bacterial cells from desiccation, heavy metals or other environmental stresses, including hostimmune responses, and to produce biofilms, thus enhancing the cells chances of colonising special ecological niches. One of the most important stress factor is salt stress for microorganisms. The present investigation is aimed to determine correlation between salt resistance and EPS production by three cyanobacterial isolates (Synechocystis sp. BASO444, Synechocystis sp. BASO507 and Synechocystis sp. BASO511). It is also aimed to investigate the effect of salt concentrations on EPS production by cyanobacteria and effect of salt on monosaccharide composition of EPS. MATERIALS AND METHODS: Cyanobacterial isolates were identified by 16 S rRNA analysis. Its salt (NaCl) tolerance and association with exopolysaccharides (EPSs) production in three cyanobacterial isolates were investigated. Also, EPS was analysed by HPLC for monomer characterization. RESULTS: Increased EPS production was associated with NaCl tolerance. The most tolerant isolate, Synechocystis sp. BASO444, secreted the most EPS (500 mg/L). EPS production by Synechocystis sp. BASO444, Synechocystis sp. BASO507 and Synechocystis sp. BASO511 was investigated following exposure to 0.2 and 0.4 M NaCl. Also, flasks containing medium without NaCl were inoculated in the same manner to serve as controls. The monosaccharide compositions of EPS produced by the three isolates following exposure to 0.2 M NaCl were analysed by HPLC. Control EPS of BASO444 was composed of glucose (97%) and galacturonic acid (3%). The composition of BASO511 (control) was glucose (95%), xylose (4.80%), arabinose (0.13%), glucuronic acid (0.03%) and galacturonic acid (0.04%). However, the composition of BASO507 (control) was glucose (0.98%), xylose (98.00%), arabinose (1.00%), glucuronic acid (0.01%) and galacturonic acid (0.01%). In the presence of 0.2 M NaCl, EPS compositions and ratios of three cyanobacterial isolates changed. DISCUSSION: Although hyperproduction of EPS in response to starvation, antiviral activity, thickening agent and cosmetic industry for product formulations has been reported for cyanobacteria, the effect of NaCl on EPS production in cyanobacteria is not a popular area of study. There are no clear reports correlating EPS production and NaCl tolerance. The gap in the data about the effect of NaCl on cyanobacterial EPS production was filled by this investigation, and the results of our study have important implications in both the industrial and environmental arenas. CONCLUSIONS: Our results indicate that 1) exposure to elevated concentrations of NaCl affects the composition of EPS produced by Synechocystis sp. BASO444, Synechocystis sp. BASO507 and Synechocystis sp. BASO511, and 2) there is a correlation between NaCl tolerance and EPS production in some cyanobacteria. RECOMMENDATIONS AND PERSPECTIVES: Differences in the monosaccharide composition and ratios of EPS may promote NaCl tolerance in these microorganisms. As well, these alternative composition polysaccharides may be important for industrial applications.

Cadmium(II) sequestration characteristics by two isolates of Synechocystis sp. in terms of exopolysaccharide (EPS) production and monomer composition.

We investigated cadmium(II) resistance and its association with exopolysaccharide (EPS) production in cyanobacteria. Increased EPS production was associated with Cd(II) resistance. The most resistant isolate, Synechocystis sp. BASO670, secreted the greatest amount of EPS (548 mg/L). EPS production by Synechocystis sp. BASO670 and Synechocystis sp. BASO672 was increased following exposure to 15 and 35 ppm Cd(II). Monomer composition of EPS belonging to each isolate was changed after Cd(II) treatment. Uronic acid contents of Cd(II) treated cells were higher than control cells of each isolate. Scanning electron microscopy (SEM) and energy dispersive X-ray spectroscopy (EDS) analysis confirmed that a considerable amount of metals had precipitated on the cell surface. Fourier transform infrared (FT-IR) spectrum analysis of EPSs belonging to both isolates indicated the presence of C-H and C-O group, which may serve as binding sites for divalent cations.

Evaluation of chromium(VI) removal behaviour by two isolates of Synechocystis sp. in terms of exopolysaccharide (EPS) production and monomer composition.

Chromium(VI) removal and its association with exopolysaccharide (EPS) production in cyanobacteria were investigated. Synechocystis sp. BASO670 produced higher EPS (548 mg L(-1)) than Synechocystis sp. BASO672 (356 mg L(-1)). While the EC(50) of the Cr(VI) for Synechocystis sp. BASO670 and Synechocystis sp. BASO672 were determined as 11.5 mg L(-1), and 2.0 mg L(-1), respectively, there was no relation between Cr(VI) removal and EPS production. Synechocystis sp. BASO672, which has higher EPS value, removed (33%) more Cr(VI) than Synechocystis sp. BASO670. Monomer compositions of EPS of each of the isolates were determined differently. Synechocystis sp. BASO672 which removed higher Cr(VI), had higher values of uronic acid and glucuronic acid (192 microg/mg and 89%, respectively). Our results showed that EPS might play a role in Cr(VI) tolerance. Monomer composition, especially uronic acid and glucuronic acid content of EPS may have enhanced Cr(VI) removal.

Characterization of vaginal lactobacilli coaggregation ability with Escherichia coli.

The coaggregation abilities of probiotic strains might enable it to form a barrier that prevents colonization by pathogenic bacteria. In the present study, the characterization of the coaggregation ability of 19 vaginal lactobacilli was studied. Coaggregation ability of all lactobacilli with Escherichia coli ATCC 11229 was positive. Only the highest coaggregation percentage of Lactobacillus acidophilus S1 was obtained with E. coli ATCC 11229 under both aerobic (71%) and anaerobic conditions (62%). The coaggregation abilities of strains occurred higher at acidic pH than at basic pH values. Moreover, the coaggregation abilities of tested strains against E. coli decreased after heat treatment (70 or 85 degrees C). Also, the relationship between hydrophobicity and coaggregation of strains was found to be significant. The effect of sonication, some enzymes (lipase and pepsin) and sodium periodate on coaggregation ability of L. acidophilus S1, which is one of the highest potentials on coaggregation ability, was investigated. Sodium periodate did not have a significant effect on coaggregation ability of L. acidophilus S1. The sonicated cell showed lower coaggregation than the control, the supernatant fluid of this sonicated cells showed similar coaggregation ability to the control. Coaggregation abilities of bacteriotherapeutic lactobacilli with pathogenic bacteria can be used for preliminary screening in order to identify potentially probiotic bacteria suitable for human use against urogenital tract infections.

Antimicrobial and antioxidant activities of Russula delica Fr.

Russula delica Fr. is a well known macrofungi which is used as a food in Turkey. The ethanolic extract of R. delica exhibited antimicrobial activity against some of the tested foodborne and spoilage bacteria. The phenolic composition of R. delica ethanolic extract was analyzed by high performance liquid chromatography (HPLC). The major component in R. delica ethanolic extract was catechin (5.33 mg/L). Antioxidant activities of the ethanolic extract of R. delica was evaluated by using 2,2-diphenyl-1-picrylhydrazyl (DPPH) radicals scavenging and chelating ability on ferrous ions assays. Scavenging effect on DPPH radicals was 26% at 10mg/ml and chelating effects on ferrous ions was 58% at 5mg/ml. In addition, the amounts of total phenol content (6.23 mg/g), ascorbic acid (2.93 mg/g), beta-carotene (0.11 mg/g) and lycopene (0.03 mg/g) in the macrofungi ethanolic extract were determined.