Screening of cyanobacteria and microalgae for their ability to synthesize silver nanoparticles with antibacterial activity

The aim of this study was to assess the ability of selected strains of cyanobacteria and microalgae to biosynthesize silver nanoparticles (Ag-NPs) by using two procedures; (i) suspending the live andwashed biomass of microalgae and cyanobacteria into the AgNO3 solution and (ii) by adding AgNO3 into a cellfree culture liquid. Ag-NPs were biosynthesized by 14 out of 16 tested strains. In most of the cases Ag-NPs were formed both in the presence of biomass as well as in the cell-free culture liquid. This indicates that the process of Ag-NPs formation involves an extracellular compound such as polysaccharide. TEM analysis showed that the nanoparticles were embedded within an organic matrix. Ag-NPs varied in shape and sizes that ranged between 13 and 31 nm, depending on the organism used. The antibacterial activity of Ag-NPs was confirmed in all but one strain of cyanobacterium (Limnothrix sp. 37-2-1) which formed the largest particles

Effects of Cyanobacterial Lipopolysaccharides from Microcystis on Glutathione-Based Detoxification Pathways in the Zebrafish (Danio rerio) Embryo

Cyanobacteria (“blue-green algae”) are recognized producers of a diverse array of toxic secondary metabolites. Of these, the lipopolysaccharides (LPS), produced by all cyanobacteria, remain to be well investigated. In the current study, we specifically employed the zebrafish (Danio rerio) embryo to investigate the effects of LPS from geographically diverse strains of the widespread cyanobacterial genus, Microcystis, on several detoxifying enzymes/pathways, including glutathione-S-transferase (GST), glutathione peroxidase (GPx)/glutathione reductase (GR), superoxide dismutase (SOD), and catalase (CAT), and compared observed effects to those of heterotrophic bacterial (i.e., E. coli) LPS. In agreement with previous studies, cyanobacterial LPS significantly reduced GST in embryos exposed to LPS in all treatments. In contrast, GPx moderately increased in embryos exposed to LPS, with no effect on reciprocal GR activity. Interestingly, total glutathione levels were elevated in embryos exposed to Microcystis LPS, but the relative levels of reduced and oxidized glutathione (i.e., GSH/GSSG) were, likewise, elevated suggesting that oxidative stress is not involved in the observed effects as typical of heterotrophic bacterial LPS in mammalian systems. In further support of this, no effect was observed with respect to CAT or SOD activity. These findings demonstrate that Microcystis LPS affects glutathione-based detoxification pathways in the zebrafish embryo, and more generally, that this model is well suited for investigating the apparent toxicophore of cyanobacterial LPS, including possible differences in structure-activity relationships between heterotrophic and cyanobacterial LPS, and teleost fish versus mammalian systems

Cyanobacterial Toxins as Allelochemicals with Potential Applications as Algaecides, Herbicides and Insecticides

Cyanobacteria (“blue-green algae”) from marine and freshwater habitats are known to produce a diverse array of toxic or otherwise bioactive metabolites. However, the functional role of the vast majority of these compounds, particularly in terms of the physiology and ecology of the cyanobacteria that produce them, remains largely unknown. A limited number of studies have suggested that some of the compounds may have ecological roles as allelochemicals, specifically including compounds that may inhibit competing sympatric macrophytes, algae and microbes. These allelochemicals may also play a role in defense against potential predators and grazers, particularly aquatic invertebrates and their larvae. This review will discuss the existing evidence for the allelochemical roles of cyanobacterial toxins, as well as the potential for development and application of these compounds as algaecides, herbicides and insecticides, and specifically present relevant results from investigations into toxins of cyanobacteria from the Florida Everglades and associated waterways

Epidemiology of recreational exposure to freshwater cyanobacteria – an international prospective cohort study

BACKGROUND: Case studies and anecdotal reports have documented a range of acute illnesses associated with exposure to cyanobacteria and their toxins in recreational waters. The epidemiological data to date are limited; we sought to improve on the design of some previously conducted studies in order to facilitate revision and refinement of guidelines for exposure to cyanobacteria in recreational waters. METHODS: A prospective cohort study was conducted to investigate the incidence of acute symptoms in individuals exposed, through recreational activities, to low (cell surface area <2.4 mm(2)/mL), medium (2.4–12.0 mm(2)/mL) and high (>12.0 mm(2)/mL) levels of cyanobacteria in lakes and rivers in southeast Queensland, the central coast area of New South Wales, and northeast and central Florida. Multivariable logistic regression analyses were employed; models adjusted for region, age, smoking, prior history of asthma, hay fever or skin disease (eczema or dermatitis) and clustering by household. RESULTS: Of individuals approached, 3,595 met the eligibility criteria, 3,193 (89%) agreed to participate and 1,331 (37%) completed both the questionnaire and follow-up interview. Respiratory symptoms were 2.1 (95%CI: 1.1–4.0) times more likely to be reported by subjects exposed to high levels of cyanobacteria than by those exposed to low levels. Similarly, when grouping all reported symptoms, individuals exposed to high levels of cyanobacteria were 1.7 (95%CI: 1.0–2.8) times more likely to report symptoms than their low-level cyanobacteria-exposed counterparts. CONCLUSION: A significant increase in reporting of minor self-limiting symptoms, particularly respiratory symptoms, was associated with exposure to higher levels of cyanobacteria of mixed genera. We suggest that exposure to cyanobacteria based on total cell surface area above 12 mm(2)/mL could result in increased incidence of symptoms. The potential for severe, life-threatening cyanobacteria-related illness is likely to be greater in recreational waters that have significant levels of cyanobacterial toxins, so future epidemiological investigations should be directed towards recreational exposure to cyanotoxins

Antibacterial Activity of Marine and Black Band Disease Cyanobacteria against Coral-Associated Bacteria

Black band disease (BBD) of corals is a cyanobacteria-dominated polymicrobial disease that contains diverse populations of heterotrophic bacteria. It is one of the most destructive of coral diseases and is found globally on tropical and sub-tropical reefs. We assessed ten strains of BBD cyanobacteria, and ten strains of cyanobacteria isolated from other marine sources, for their antibacterial effect on growth of heterotrophic bacteria isolated from BBD, from the surface mucopolysaccharide layer (SML) of healthy corals, and three known bacterial coral pathogens. Assays were conducted using two methods: co-cultivation of cyanobacterial and bacterial isolates, and exposure of test bacteria to (hydrophilic and lipophilic) cyanobacterial cell extracts. During co-cultivation, 15 of the 20 cyanobacterial strains tested had antibacterial activity against at least one of the test bacterial strains. Inhibition was significantly higher for BBD cyanobacteria when compared to other marine cyanobacteria. Lipophilic extracts were more active than co-cultivation (extracts of 18 of the 20 strains were active) while hydrophilic extracts had very limited activity. In some cases co-cultivation resulted in stimulation of BBD and SML bacterial growth. Our results suggest that BBD cyanobacteria are involved in structuring the complex polymicrobial BBD microbial community by production of antimicrobial compounds

Effects of Emulsifying Agents on the Microstructure and Other Characteristics of Process Cheese - A Review

Sodium phosphates , polyphosphates, and citrates are melting salts (emulsifying agents) most commonly used in the manufacture of process cheese either alone or in mixtures . Their role during processing is to sequester calcium in the natural cheese, to solubilize protein and increase its hydration and swelling, to facilitate emul sification of fat , and to adjust and stabilize pH. Changes taking place in natural cheese during processing can be studied by microscopy. Micrographs demonstrating the emulsification of fat , presence of salt crystals, and partial solubilization of protein in labo ratory- made and commercial process cheeses have been used to illustrate the various effects of the emulsifying agents. Optical, particularly polarizing and fluorescence microscopy provides rapid information. Electron microscopy reveals greater detai 1. In combination with energy dispersive spectrometry, electron microscopy can be used to analyze the chemical composition of salt crystals in the process cheese. However, detailed studies of the relationships existing among the microstructure of the process cheese, its composition , and physical properties such as consistency, spreadability, capability of remelting etc. have yet to be carried out

Occurrence and distribution of novel botryococcene hydrocarbons in freshwater wetlands of the Florida Everglades

A high abundance of isoprenoid hydrocarbons, the botryococcenes, with carbon numbers from 32 to 34 were detected in the Florida Everglades freshwater wetlands. These compounds were present in varying amounts up to 106 μg/gdw in periphyton, 278 μg/gdw in floc, and 46 μg/gdw in soils. Their structures were determined based on comparison to standards, interpretation of their mass spectra and those of their hydrogenation products, and comparison of Kovats indexes to those reported in the literature. A total of 26 cyclic and acyclic botryococcenes with 8 skeletons were identified, including those with fewer degrees of unsaturation, which are proposed as early diagenetic derivatives from the natural products. This is the first report that botryococcenes occur in the Everglades freshwater wetlands. Their potential biogenetic sources from green algae and cyanobacteria were examined, but neither contained botryococcenes. Thus, the source implication of botryococcenes in this ecosystem needs further study

Quantifying the responses of calcareous periphyton crusts to rehydration: A microcosm study (Florida Everglades)

We examined the high-resolution temporal dynamics of recovery of dried periphyton crusts following rapid rehydration in a phosphorus (P)-limited short hydroperiod Everglades wetland. Crusts were incubated in a greenhouse in tubs containing water with no P or exogenous algae to mimic the onset of the wet season in the natural marsh when heavy downpours containing very low P flood the dry wetland. Algal and bacterial productivity were tracked for 20 days and related to compositional changes and P dynamics in the water. A portion of original crusts was also used to determine how much TP could be released if no biotic recovery occurred. Composition was volumetrically dominated by cyanobacteria (90%) containing morphotypes typical of xeric environments. Algal and bacterial production recovered immediately upon rehydration but there was a net TP loss from the crusts to the water in the first 2 days. By day 5, however, cyanobacteria and other bacteria had re-absorbed 90% of the released P. Then, water TP concentration reached a steady-state level of 6.6 μg TP/L despite water TP concentration through evaporation. Phosphomonoesterase (PMEase) activity was very high during the first day after rehydration due to the release of a large pre-existing pool of extracellular PMEase. Thereafter, the activity dropped by 90% and increased gradually from this low level. The fast recovery of desiccated crusts upon rehydration required no exogenous P or allogenous algae/bacteria additions and periphyton largely controlled P concentration in the water

Quantifying the responses of calcareous periphyton crusts to rehydration: A microcosm study (Florida Everglades)

We examined the high-resolution temporal dynamics of recovery of dried periphyton crusts following rapid rehydration in a phosphorus (P)-limited short hydroperiod Everglades wetland. Crusts were incubated in a greenhouse in tubs containing water with no P or exogenous algae to mimic the onset of the wet season in the natural marsh when heavy downpours containing very low P flood the dry wetland. Algal and bacterial productivity were tracked for 20 days and related to compositional changes and P dynamics in the water. A portion of original crusts was also used to determine how much TP could be released if no biotic recovery occurred. Composition was volumetrically dominated by cyanobacteria (90%) containing morphotypes typical of xeric environments. Algal and bacterial production recovered immediately upon rehydration but there was a net TP loss from the crusts to the water in the first 2 days. By day 5, however, cyanobacteria and other bacteria had re-absorbed 90% of the released P. Then, water TP concentration reached a steady-state level of 6.6 μg TP/L despite water TP concentration through evaporation. Phosphomonoesterase (PMEase) activity was very high during the first day after rehydration due to the release of a large pre-existing pool of extracellular PMEase. Thereafter, the activity dropped by 90% and increased gradually from this low level. The fast recovery of desiccated crusts upon rehydration required no exogenous P or allogenous algae/bacteria additions and periphyton largely controlled P concentration in the water

Characterization of Roseofilum reptotaenium (Oscillatoriales, Cyanobacteria) gen. et sp. nov. isolated from Caribbean Black band disease

Black band disease (BBD) is a pathogenic microbial consortium dominated in terms of biomass by phycoerythrin-rich, gliding, filamentous cyanobacteria. Studies of BBD using molecular methods have shown that the 16S rRNA gene sequence of \u27Oscillatoria\u27 sp. is consistently found in BBD samples worldwide. The purpose of this study is to erect a new genus, Roseofilum gen. et sp. nov., encompassing these and other previously reported strains. Using a polyphasic approach we characterized two strains of BBD Oscillatoria isolated from BBD infected corals in the Caribbean. These strains have the ability to tolerate sulphide, anoxia, and pH values in the range from 4.5 to 10, with optimal growth at pH 6 to 8. Growth occurs by photoautotrophy, including sulphide-insensitive oxygenic photosynthesis, and mixotrophy but not by heterotrophy under dark aerobic or anaerobic conditions. Both strains synthesize microcystin-LR. Results of infectivity experiments carried out under controlled laboratory conditions showed that both strains are capable of initiating an infection on healthy coral that resembles in situ BBD. 16S rRNA gene sequence data place these strains into a highly supported clade with other strains identified as Oscillatoria sensu lato, yet clearly genetically distinct from the type, Oscillatoria princeps. Further, while the BBD strains share more morphological similarity with members of Phormidium, they are also distinct from this genus based on sequence data. Based on morphology, ecology, physiology, and phylogenetic distinctiveness, we propose the novel epithet Roseofilum reptotaenium