Autofluorescence is a remarkable property of human skin. It can be excited by UV and observed in the dark using special detection systems. The method of fluorescence photography (FP) is an effective non-invasive tool for skin assessment. It involves image capturing by a camera the emission of light quanta from fluorophore molecules in the skin. It serves as a useful tool for cosmetic and skincare research, especially for the detection of pathological skin states, like acne, psoriasis, etc. To the best of our knowledge, there is currently no comprehensive review that fully describes the application and physical principles of FP over the past five years. The current review covers various aspects of the skin FP method from its biophysical basis and the main fluorescent molecules of the skin to its potential applications and the principles of FP recording and analysis. We pay particular attention to recently reported works on the automatic analysis of FP based on artificial intelligence (AI). Thus, we argue that FP is a rapidly evolving technology with a wide range of potential applications. We propose potential directions of the development of this method, including new AI algorithms for the analysis and expanding the range of applications.
Microalgae are the richest source of natural carotenoids, which are valuable pigments with a high share of benefits. Often, carotenoid-producing algae inhabit specific biotopes with unfavorable or even extremal conditions. Such biotopes, including alpine snow fields and hypersaline ponds, are widely distributed in Europe. They can serve as a source of new strains for biotechnology. The number of algal species used for obtaining these compounds on an industrial scale is limited. The data on them are poor. Moreover, some of them have been reported in non-English local scientific articles and theses. This review aims to summarize existing data on microalgal species, which are known as potential carotenoid producers in biotechnology. These include Haematococcus and Dunaliella, both well-known to the scientific community, as well as less-elucidated representatives. Their distribution will be covered throughout Europe: from the Greek Mediterranean coast in the south to the snow valleys in Norway in the north, and from the ponds in Amieiro (Portugal) in the west to the saline lakes and mountains in Crimea (Ukraine) in the east. A wide spectrum of algal secondary carotenoids is reviewed: β-carotene, astaxanthin, canthaxanthin, echinenone, adonixanthin, and adonirubin. For convenience, the main concepts of biology of carotenoid-producing algae are briefly explained.
A novel chlorophyte algae strain with outstanding resilience to high inorganic phosphate (Pi) concentrations in the medium was isolated from a phosphorus-polluted site near a rock phosphate mine. According to the morphological, ultrastructural and genetic criteria the strain was assigned to the species Micractinium simplicissimum H.Chae, H.-G. Choi & J.H.Kim. This strain retained cell viability and growth capacity in the presence of Pi concentrations up to 14 g L–1. The uptake of Pi by the cells was moderate (equal to ca. 0.7% increase in cell dry weight P percentage) regardless of the amount of the exogenic Pi added to the culture. At the same time, approximately a half of the Pi removed by the M. simplicissimum from the culture was reversely adsorbed by the cell surface and/or the intercellular matrix and cell debris. The ultrastructural studies indicated the metabolically active status of the cells together with the presence of phosphorus-rich (likely, polyphosphate) inclusions outside and inside the cells (mainly in vacuoles). We hypothesized that the Pi resilience of the studied strain stems from its high Pi adsorption capacity together with its ability to throttle the Pi influx into the cell preventing the rapid buildup of intracellular Pi and potentially toxic short-chain polyphosphate.
UV-A is the main ultraviolet component of natural (solar) radiation. Despite it, its effect on phototrophs is studied less than UV-B. Effects of UV-A on photosynthetic apparatus of three carotenoid-producing microalgae were elucidated. Photosynthetic activity was studied using chlorophyll fluorescence analysis. Cell extracts were evaluated by absorbance spectroscopy. On the one hand, there were some common features of three strains. In all cases the changes involved PSII primary photochemistry and antennae size. All strains accumulated UV-absorbing polar compounds. On the other hand, some responses were different. Upregulation of non-photochemical quenching was observed only in B. aggregatus BM5/15, whereas in other cases its level was low. H. rubicundus BM7/13 and Deasonia sp. NAMSU 934/2 accumulated secondary carotenoids, whereas B. aggregatus BM5/15 accumulated primary ones. Microscopic features of the cultures were also different. H. rubicundus BM7/13 and Deasonia sp. NAMSU 934/2 were represented mostly by solitaire cells or small cell clusters, lacking their green color; the cells of B. aggregatus BM5/15 formed aggregates from green cells. Cell aggregation could be considered as an additional UV-protecting mechanism. Finally, the strains differed by their viability. B. aggregatus BM5/15 was most resistant to UV-A, whereas massive cell death was observed in two other cultures.
The results of the complex multimodal optical study utilizing Raman and fluorescence lifetime imaging of the cells of micro alga Bracteacoccus aggregatus BM5/15 are presented.
Fuel (especially kerosene) biodamage is a challenge for global industry. In aviation, where kerosene is a widely used type of fuel, its biodeterioration leads to significant damage. Six isolates of micromycetes from the TS-1 aviation kerosene samples were obtained. Their ability to grow on the fuel was studied, and the difference between biodegradation ability was shown. Micromycetes belonged to the Talaromyces, Penicillium, and Aspergillus genera. It was impossible to obtain bacterial isolates associated with their mycelium. However, 16S rRNA metabarcoding and microscopic observations revealed the presence of bacteria in the micromycete isolates. It seems to be that kerosene-degrading fungi were associated with uncultured bacteria. Proteobacteria, Actinobacteria, Bacteroidetes, and Firmicutes were abundant in the fungal cultures isolated from the TS-1 jet fuel samples. Most genera among these phyla are known as hydrocarbon degraders. Only bacteria-containing micromycete isolates were able to grow on the kerosene. Most likely, kerosene degradation mechanisms are based on synergism of bacteria and fungi.
Carotenoids astaxanthin and β-carotene are widely used natural antioxidants. They are key components of functional food, cosmetics, drugs and animal feeding. They hold leader positions on the world carotenoid market. In current work, we characterize the new strain of the green microalga Bracteacoccus aggregatus BM5/15 and propose the method of its culturing in a bubble-column photobioreactor for simultaneous production of astaxanthin and β-carotene. Culture was monitored by light microscopy and pigment kinetics. Fatty acid profile was evaluated by tandem gas-chromatography–mass spectrometry. Pigments were obtained by the classical two-stage scheme of autotrophic cultivation. At the first, vegetative, stage biomass accumulation occurred. Maximum specific growth rate and culture productivity at this stage were 100–200 mg∙L−1∙day−1, and 0.33 day−1, respectively. At the second, inductive, stage carotenoid synthesis was promoted. Maximal carotenoid fraction in the biomass was 2.2–2.4%. Based on chromatography data, astaxanthin and β-carotene constituted 48 and 13% of total carotenoid mass, respectively. Possible pathways of astaxanthin synthesis are proposed based on carotenoid composition. Collectively, a new strain B. aggregatus BM5/15 is a potential biotechnological source of two natural antioxidants, astaxanthin and β-carotene. The results give the rise for further works on optimization of B. aggregatus cultivation on an industrial scale.
The microalga Coelastrella rubescens dwells in habitats with excessive solar irradiation; consequently, it must accumulate diverse compounds to protect itself. We characterized the array of photoprotective compounds in C. rubescens. Toward this goal, we exposed the cells to high fluxes of visible light and UV-A and analyzed the ability of hydrophilic and hydrophobic extracts from the cells to absorb radiation. Potential light-screening compounds were profiled by thin layer chromatography and UPLC-MS. Coelastrella accumulated diverse carotenoids that absorbed visible light in the blue–green part of the spectrum and mycosporine-like amino acids (MAA) that absorbed the UV-A. It is the first report on the occurrence of MAA in Coelastrella. Two new MAA, named coelastrin A and coelastrin B, were identified. Transmission electron microscopy revealed the development of hydrophobic subcompartments under the high light and UV-A exposition. We also evaluate and discuss sporopollenin-like compounds in the cell wall and autophagy-like processes as the possible reason for the decrease in sunlight absorption by cells, in addition to inducible sunscreen accumulation. The results suggested that C. rubescens NAMSU R1 accumulates a broad range of valuable photoprotective compounds in response to UV-A and visible light irradiation, which indicates this strain as a potential producer for biotechnology.
Haematococcus lacustris is a natural source of a valuable ketocarotenoid astaxanthin. Under autotrophic growth conditions, it exists in the form of a community with bacteria. The close coexistence of these microorganisms raises two questions: how broad their diversity is and how they interact with the microalga. Despite the importance these issues, little is known about microorganisms existing in Haematococcus cultures. For the first time, we characterize the dynamic of the H. lacustris microbiome of the microbiome of Haematococcus (a changeover of the bacterial associated species as function of the time) cultivated autotrophically in a photobioreactor based on 16S rRNA metabarcoding data. We found that Proteobacteria and Bacteroidetes are predominant phyla in the community. The Caulobacter bacterium became abundant during astaxanthin accumulation. These data were supported by microscopy. We discuss possible roles and interactions of the community members. These findings are of potential significance for biotechnology. They provide an insight into possible bacterial contamination in algal biomass and reveal the presence of bacteria essential for the algal growth.
To date, it has been increasingly clear, that microbiota plays essential role in the host microorganism. Due to high phylogenetic diversity and wide distribution understanding of gut microbiota of fish is ambiguous. We elucidate previously undescribed diversity of bacteria in the White Sea fish posterior intestine based on 16S rRNA metabarcoding: shorthorn sculpin (Myoxocephalus scorpius), lumpfish (Cyclopterus lumpus) arctic flounder (Liopsetta glacialis), cod (Gadus morhua), and herring (Clupea pallasii). Proteobacteria were most abundant in all the samples. All datasets contained high number of reads corresponded to Psychrobacter, Halomonas, Pseudoalteromonas and Vibrio abundant in the White Sea water.Vast majority of bacterial reads corresponded to the Vibrionaceae (Aliivibrio, Vibrio, and Photobacterium). It is in accordance with our previous study focused on bioluminescent bacteria, where we isolated strains of these genera from the same fish intestine samples. The datasets of G. morhua and Cl. pallasii were enriched by reads of Shewanella (also isolated previously microbiologically due to luminescence of some its strains); the datasets of G. morhua and M. scorpius contained high number of Mycoplasma, which is in line with previous reports on the microbiome of marine fish gut from other geographical regions. Collectively, on the one hand, the study is a confirmation of known data on marine fish gut microbiome, on the other hand, it reveals some specific patterns of White Sea fish gut microbiota, e.g. presence of bacteria common for water from this region.
The biodiversity of microbial communities from phosphorus-polluted sites from Northern and Southern regions was investigated, groups of microorganisms with biotechnological potential were detected. According to the results, samples from Southern regions were characterized by lower biodiversity than the northern ones.
Bioluminescence is a spectacular feature of some prokaryotes. In the present work, we address the distribution of bioluminescence among bacteria isolated from the White Sea finfishes. Luminous bacteria are widely distributed throughout the World Ocean. Many strains have been isolated and described for tropical latitudes, while Nordic seas still remain quite a white spot in studying bioluminescence of bacteria. We describe the strains related to the two main genera of luminous bacteria, Photobacterium and Aliivibrio, as well as Shewanella and Vibrio. They are related to families Vibrionaceae and Shewanellaceae of the Gammaproteobacteria class. Here, we at the first time, report the bioluminescence of the Enterobacteriaceae Kosakonia cowanii. Moreover, we applied the polyphasic approach to identify and describe the isolated microorganisms. The data on sequencing, diversity of cell fine structure, and light emission spectra at room temperature on the solid medium are discussed. The bacteria are characterized by features in their light emission spectra. It may reflect possible molecular mechanisms of bioluminescence as well as features of bacterial composition. The obtained data expands the existing body of knowledge about the bioluminescence spread among the bacteria of Nordic latitudes and provides complex information that is crucial for their precise identification.
The decrease of the available phosphorus reserves has a detrimental effect on agriculture and other industrial sectors. Various microorganisms are used in biotechnology for bioremoval of the inorganic phosphate from wastewaters, with the following application as biofertilizers for agricultural crops production. Both associations and monocultures of microalgae, cyanobacteria, and heterotrophic bacteria are able to accumulate high concentrations of phosphorus in their cells and are useful microorganisms for biotechnological phosphorus uptake. Communities of microorganisms formed in water bodies near tailing sites of the apatite mines were investigated. We studied the communities formed in the Lake Bolshaya Imandra (Murmansk Oblast, Karelia, Russia), in the tailing pond, as well as in a temporary pond (puddle); in total, five samples were studied. The morphological characteristics and the taxonomic composition of the samples were studied by light microscopy and DNA metabarcoding. Among bacteria, representatives of the phyla Proteobacteria, Firmicutes, and Actinobacteria were identified as dominants; bacteria of the phylum Bacteroidetes were less present. The results revealed a high degree of similarity of communities in terms of taxonomic structure at the genus level, which indicates the formation of common unique conditions in this region. The smallest variety was noted in the temporary puddle pond, which indicates less stability of the community formed in it.
Carotenogenic microalgae are unicellular photosynthetic organisms with the ability to accumulate carotenoids. Carotenoid accumulation is a protective reaction against environmental stress factors, such as bright light and extreme temperatures. It makes the survival of these microorganisms under harsh environmental conditions possible. The diversity of carotenogenic microalgae has been described in detail for Central Europe and North America, as well as for tropical and subtropical latitudes with relatively favorable environments. However, data about these microorganisms in polar and subpolar latitudes are scarce and restricted to few reports. We isolated several strains of carotenogenic microalgae from the coastal zone of the White Sea, where they were abundant. The obtained microalgae related to four species of Chlorophytes: Haematococcus lacustris, H. rubicundus, Coelastrella aeroterrestrica and Bracteacoccus aggregatus. The last three species have been reported for polar latitudes for the first time. Most likely, carotenogenic algae in the White Sea coast are abundant due to their high physiological and metabolic plasticity, which is essential for surviving under adverse conditions of the northern regions. Pigment composition of the strains is provided. Their predominant carotenoids were astaxanthin and beta-carotene. Further, the obtained strains may be considered as potential producers of natural pigments for biotechnology.
The microalga Haematococcus lacustris (formerly H. pluvialis) is the richest source of the valuable pigment astaxanthin, accumulated in red aplanospores (haematocysts). In this work, we report on the photoprotective mechanisms in H. lacustris, conveying this microalga its ability to cope with a wide range of adverse conditions, with special emphasis put on non-photochemical quenching (NPQ) of the excited chlorophyll states. We studied the changes in the primary photochemistry of the photosystems (PS) as a function of irradiance and the physiological state. We leveraged the transcriptomic data to gain a deeper insight into possible NPQ mechanisms in this microalga. Peculiar to H. lacustris is a bi-phasic pattern of changes in photoprotection during haematocyst formation. The first phase coincides with a transient rise of photosynthetic activity. Based on transcriptomic data, high NPQ level in the first phase is maintained predominantly by the expression of PsbS and LhcsR proteins. Then, (in mature haematocysts), stress tolerance is achieved by optical shielding by astaxanthin and dramatic reduction of photosynthetic apparatus. In contrast to many microalgae, shielding plays an important role in H. lacistris haematocysts, whereas regulated NPQ is suppressed. Astaxanthin is decoupled from the PS, hence the light energy is not transferred to reaction centers and dissipates as heat. It allows to retain a higher photochemical yield in haematocysts comparing to vegetative cells. The ability of H. lacustris to substitute the "classical" active photoprotective mechanisms such as NPQ with optic shielding and general metabolism quiescence makes this organism a useful model to reveal photoprotection mechanisms.
The libraries of bacterial 16SrRNA gene fragment from algal-bacterial consortia of the White Sea coastal zone are analyzed. Up to 3% of the reads have revealed to correspond to eukaryotic rRNA. They related to following main eukaryotic clades: Discoba, Stramenopiles, Ciliata, Amoebozoa, and Nucletmycea. Amoebae, especially Vermamoeba, were present in all samples. In one sample, heterolobose amoeba Paravahlkampfia was detected. These microorganisms are parasites of microalgae, which can induce significant damage to industrial cultures. However, the data on their physiology and distribution are scarce. This study provides new evidence about the diversity of herbivorous eukaryotic microorganisms in natural algal-containing consortia.
Haematococcus lacustris is a biotechnologically important green unicellular alga producing widely used keta-karotenoid astaxanthin. In natural habitats, it exists in the form of algal-bacterial community, and under laboratory conditions, it is also accompanied by bacteria. The issue of the bacterial composition of industrial algal cultures is widely recognized as important. However, there is a dearth of information about bacterial composition of H. lacustris communities. In current work, we analyze the composition of natural H. lacustris communities from the White Sea coastal temporal rock ponds. For the first time, a 16S rRNA gene-based metagenome of natural H. lacustris bacterial communities has been generated. Main results of its analysis are as follow. Bacterial families Comamonadaceae, Cytophagaceae, Xanthomonadaceae, Acetobacteraceae, Rhodobacteraceae, and Rhodocyclaceae were observed in all studied H. lacustris natural communities. They also contained genera Hydrogenophaga and Cytophaga. Bacteria from the Hydrogenophaga genus were present in H. lacustris cultures after their isolation under the conditions of laboratory cultivation. Similar to other planktonic microalgae, H. lacustris forms a phycosphere around the cells. In this zone, bacteria attached to the algal surface. The contact between H. lacustris and bacteria is maintained even after sample drying. The study provides information about possible members of H. lacustris core microbiome, which can be presented in the industrial and laboratory cultures of the microalga.
Cyanobacteria characterized by exceptional tolerance to environmental stresses often become pioneer settlers in habitats with harsh conditions. There, they can constitute the core of microbial communities. The taxonomic composition of the cyanobacterial component of algal–bacterial consortia dwelling in a habitat with particularly harsh conditions (rock baths at the coast of Kandalaksha Bay of the White Sea) has been elucidated for the first time. Two workflows of the taxonomic analysis of the cyanobacteria were tested including the combinations of two programs and two databases (QIIME+Greengenes and Usearch+NCBI GenBank). Our results obviated the need of the using of a complex approach combining morphological and metagenomic analyses for revealing the taxonomic structure of cyanobacteria in natural habitats. Our results show that the cyanobacterial component of the consortia from the habitats with harsh and highly volatile environmental conditions is enriched with non-diazotrophic and diazotrophic non-branched filamentous cyanobacteria.