Preliminary Study on Microplastic Contamination in Black Sea Cetaceans: Gastrointestinal Analysis of Phocoena phocoena relicta and Tursiops truncatus ponticus.

Plastic pollution is a global concern that has a significant impact on marine life. Plastic is widely used and has become a pervasive pollutant in marine environments. Plastic contamination has been documented both in marine environments and biota. Plastic contamination in cetacean gastro-intestinal tract (GIT) content has received limited attention, especially in the Black Sea. This study aims to investigate plastic contamination in the GITs of bottlenose dolphins and harbour porpoises, introducing a novel methodology. Given the limited exploration of this issue in the Black Sea, the research predominantly focuses on microplastic contamination. The GITs were sampled through necropsy from stranded and by-caught cetaceans, and content was washed through a multi-sieves tool. The material retained on each sieve was analysed following specific protocols. All (100%) of the GITs contained plastics (meso- and microplastics). In total, 1059 items (fibres, fragments, and beads) ranging from 22.86 µm to 5776 µm were found, suggesting a high contamination level in the Black Sea cetaceans. Future efforts should concentrate on increasing the number of samples and using the results for the implementation of the Marine Strategy Framework Directive (MSFD).

Quantitative monitoring of diverse fish communities on a large scale combining eDNA metabarcoding and qPCR.

Environmental DNA (eDNA) metabarcoding is an effective method for studying fish communities but allows only an estimation of relative species abundance (density/biomass). Here, we combine metabarcoding with an estimation of the total abundance of eDNA amplified by our universal marker (teleo) using a quantitative (q)PCR approach to infer the absolute abundance of fish species. We carried out a 2850-km eDNA survey within the Danube catchment using a spatial integrative sampling protocol coupled with traditional electrofishing for fish biomass and density estimation. Total fish eDNA concentrations and total fish abundance were highly correlated. The correlation between eDNA concentrations per taxon and absolute specific abundance was of comparable strength when all sites were pooled and remained significant when the sites were considered separately. Furthermore, a nonlinear mixed model showed that species richness was underestimated when the amount of teleo-DNA extracted from a sample was below a threshold of 0.65 × 106 copies of eDNA. This result, combined with the decrease in teleo-DNA concentration by several orders of magnitude with river size, highlights the need to increase sampling effort in large rivers. Our results provide a comprehensive description of longitudinal changes in fish communities and underline our combined metabarcoding/qPCR approach for biomonitoring and bioassessment surveys when a rough estimate of absolute species abundance is sufficient.

Polymer Membranes as Innovative Means of Quality Restoring for Wastewater Bearing Heavy Metals.

The problem that has aroused the interest of this review refers to the harmful effect of heavy metals on water sources due to industrial development. In this respect, the review is aimed at achieving a literature survey on the outstanding results and advancements in membranes and membrane technologies for the advanced treatment of heavy metal-loaded wastewaters. Particular attention is given to synthetic polymer membranes, for which the proper choice of precursor material can provide cost benefits while ensuring good decontamination activity. Furthermore, it was also found that better removal efficiencies of heavy metals are achieved by combining the membrane properties with the adsorption properties of inorganic powders. The membrane processes of interest from the perspective of industrial applications are also discussed. A noteworthy conclusion is the fact that the main differences between membranes, which refer mainly to the definition and density of the pore structure, are the prime factors that affect the separation process of heavy metals. Literature studies reveal that applying UF/MF approaches prior to RO leads to a better purification performance.

Molecularly Imprinted Ligand-Free Nanogels for Recognizing Bee Venom-Originated Phospholipase A2 Enzyme.

In this study, ligand-free nanogels (LFNGs) as potential antivenom mimics were developed with the aim of preventing hypersensitivity and other side effects following massive bee attacks. For this purpose, poly (ethylene glycol) diacrylate was chosen as a main synthetic biocompatible matrix to prepare the experimental LFNGs. The overall concept uses inverse mini-emulsion polymerization as the main route to deliver nanogel caps with complementary cavities for phospholipase A2 (PLA2) from bee venom, created artificially with the use of molecular imprinting (MI) technologies. The morphology and the hydrodynamic features of the nanogels were confirmed by transmission electron microscopy (TEM) and dynamic light scattering (DLS) analysis. The following rebinding experiments evidenced the specificity of molecularly imprinted LFNG for PLA2, with rebinding capacities up to 8-fold higher compared to the reference non-imprinted nanogel, while the in vitro binding assays of PLA2 from commercial bee venom indicated that such synthetic nanogels are able to recognize and retain the targeted PLA2 enzyme. The results were finally collaborated with in vitro cell-viability experiments and resulted in a strong belief that such LFNG may actually be used for future therapies against bee envenomation.

Bacterial communities of the Black Sea exhibit activity against persistent organic pollutants in the water column and sediments.

The ability of bacteria to degrade organic pollutants influences their fate in the environment, impact on the other biota and accumulation in the food web. The aim of this study was to evaluate abundance and expression activity of the catabolic genes targeting widespread pollutants, such as polyaromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs) and hexachloro-cyclohexane (HCH) in the Black Sea water column and sediments. Concentrations of PAHs, PCBs and HCH were determined by gas chromatography (GC) coupled to mass spectrometry (MS) and electron capture (ECD) detectors. bphA1, PAH-RHDα, nahAc, linA and linB that encode biphenyl 2,3 dioxygenase, α-subunits of ring hydroxylating dioxygenases, naphthalene dioxygenase, dehydrochlorinase and halidohydrolase correspondently were quantified by quantitative PCR. More recalcitrant PAHs, PCBs and HCH tended to accumulate in the Black Sea environments. In water samples, 3- and 4-ringed PAHs outnumbered naphthalene, while PAHs with > 4 rings prevailed in the sediments. Congeners with 4-8 chlorines with ortho-position of the substituents were the most abundant among the PCBs. ß-HCH was determined at highest concentration in water samples, and total amount of HCH exceeded its legacy Environmental Quality Standard value. bphA1, was the most numerous gene in water layers (105 copies/mL) and sediments (105 copies/mg), followed by linB and PAH-RHDα genes (103 copies/mL; 105 copies/mg). The least abundant genes were linA (103 copies/mL; 104 copies/mg) and nahAc (102 copies/mL; 104 copies/mg). The most widely distributed gene bphА1 was one of the least expressed (10-3-10-2 copies/mL; 10-1 copies/mg). The most actively expressed genes were linB (101-102 copies/mL; 103 copies/mg), PAH-RHDα (101 copies/mL; 102 copies/mg) and linA (10-1-100 copies/mL; 100 copies/mg). Interaction of bacteria with PAHs, PCBs and HCH is evidenced by high copy numbers of the catabolic genes that initiate their degradation. More persistent compounds, such as high-molecular weight PAHs or ß-HCH are accumulating in the Black Sea water and sediments, albeit microbial activity is directed against them.

Correlations between periodontal indices and osteoporosis.

In the pathogenesis of chronic periodontitis, there are general systemic factors which play a major role, such as osteoporosis, with menopause as the most common etiological factor, and other pathological determining conditions for osteoporosis as well (ovary, thyroid and malignant tumors). The aim of the present study was to assess the correlations between periodontal indices and osteoporosis. The study was performed on 35 patients with periodontal disease aged between 45 and 79 years. These patients were divided into two groups: a study group with osteoporosis and periodontal disease (n=25) and a control group with periodontal disease (n=10) only. The periodontal assessment included community periodontal index (CPI), gingival inflammation index (GI), plaque index (PI), body mass index (BMI), bone mineral density (BMD), tooth mobility and tooth loss. Osteoporosis was assessed by dual-energy X-ray absorptiometry. Results were statistically analyzed with Microsoft Excel software and XLSTAT. The results showed that patients in the study group had higher values of periodontal indices, and a highly significant inverse correlation was observed between the CPI and the tooth loss. Inverse correlations between BMI and tooth mobility, as well as BMI and CPI were determined for the study group. In conclusion, the positive association between BMD and GI shows that the gingival index can be a predictive factor in the occurrence of osteoporosis.

Niche partitioning of bacterial communities along the stratified water column in the Black Sea.

The Black Sea is the largest semi-closed permanently anoxic basin on our planet with long-term stratification. The study aimed at describing the Black Sea microbial community taxonomic and functional composition within the range of depths spanning across oxic/anoxic interface, and to uncover the factors behind both their vertical and regional differentiation. 16S rRNA gene MiSeq sequencing was applied to get the data on microbial community taxonomy, and the PICRUSt pipeline was used to infer their functional profile. The normoxic zone was mainly inhabited by primary producers and heterotrophic prokaryotes (e.g., Flavobacteriaceae, Rhodobacteraceae, Synechococcaceae) whereas the euxinic zone-by heterotrophic and chemoautotrophic taxa (e.g., MSBL2, Piscirickettsiaceae, and Desulfarculaceae). Assimilatory sulfate reduction and oxygenic photosynthesis were prevailing within the normoxic zone, while the role of nitrification, dissimilatory sulfate reduction, and anoxygenic photosynthesis increased in the oxygen-depleted water column part. Regional differentiation of microbial communities between the Ukrainian shelf and offshore zone was detected as well, yet it was significantly less pronounced than the vertical one. It is suggested that regional differentiation within a well-oxygenated zone is driven by the difference in phytoplankton communities providing various substrates for the prokaryotes, whereas redox stratification is the main driving force behind microbial community vertical structure.

Holistic pelagic biodiversity monitoring of the Black Sea via eDNA metabarcoding approach: From bacteria to marine mammals.

As the largest semi-closed marine ecosystem in the world, the Black Sea has been heavily affected by human activities for a long time. Describing the biodiversity of multi-trophic biota in pelagic zone of the Black Sea and identifying the dominant environmental factors are prerequisites for protecting the sustainability of ecosystems. However, up to now, the taxonomic and distributional information about the Black Sea biota is not clear. Here, we employed a Tree-of-Life metabarcoding to analyze the biodiversity of eight communities in the Black Sea, investigated their biogeographical distribution, and further analyzed the influence of biological and abiotic factors on biota on large scales. We found that, (1) Over 8900 OTUs were detected in the Black Sea, of which 630 species were identified, covering the holistic biota from single-celled (bacteria 5620 OTUs 141 species; algae 1096 OTUs 185 species; protozoa 546 OTUs 146 species) to multicellular organisms (invertebrate metazoans 150 OTUs 34 species; fishes 1369 OTUs 76 species; large marine mammals 39 OTUs 5 species). (2) Higher trophic organisms (fishes and large mammals) distributed more evenly in space than the lower (microorganisms, protozoa and invertebrates). For lower trophic organisms, the vertical stratification was more obvious than the horizontal stratification (vertical p < 0.02, horizontal p < 0.05). (3) The bottom trophic organisms (bacteria and algae) of the food web significantly affected the distribution and composition of the others through biological interactions (Mantel p < 0.05). (4) At the level of abiotic factors, the effect of local species sorting on the composition of communities was 15% higher than that of mass dispersal effect. For the first time, this study monitored and profiled the holistic biodiversity in the pelagic zone of the Black Sea, and provided technological advances and preliminary knowledge for the ongoing Black Sea ecosystem protection efforts.