Determination of the antimicrobial activity of lactic acid bacteria isolated from the Black sea mussel Mytilus galloprovincialis Lamarck, 1819

The present study reports on the determination of the antimicrobial activity of lactic acid bacteria (LAB) isolated from the Black sea mussel Mytilus galloprovincialis Lamarck, 1819. The samples were collected in the period of August 2018 until March 2021. The BIOLOG system was used for microbiological determination. From the mussel M. galloprovincialis Lam. four species of LAB were isolated - Sporolactobacillus kofuensis, Lactobacillus sakei, Streptococcus gallolyticus ss gallolyticus and Lactibacillus brevis. The activity of the strains was determined against test cultures (Escherichia coli 3398, Staphylococus aureus 745, Bacillus subtilis 6633, Salmonella typhimurium 3591, Listeria monocytogens 863 Enterobacter aerogenes 3691, Aspergillus niger, Penicillium claviforme, Saccharomyces cerevisae, Candida albicans 8673 and Candida glabrata 72). Before the analysis for antimicrobial activity, the LAB were cultured in media with different concentrations of sugars - 2, 5 and 10%. The results showed that 4 strains S. kofuensis, L. sakei, S. gallolyticus ss gallolyticus and L. brevis cultured on glucose and oligosaccharides completely lost their activity in all studied variants. Therefore, some carbohydrates (glucose) and oligosaccharides induce the synthesis outside the cell of biologically active molecules, which can probably be attributed to peptides/proteins

Seasonal changes in the pro/antioxidant status of mussels Mytilus galloprovincialis (Lamarck, 1819) from Bulgarian Black Sea coastal habitats

The pro/antioxidant status of marine macrozoobentic organisms is being increasingly applied in environmental monitoring and conservation programs. The oxidative stress level in marine bivalves can provide valuable information not only on the health of the organisms and their populations, but also on the current state of habitats and ecosystems. The aim of the present study was to make the first comprehensive investigation of the seasonal changes in the antioxidant activity in different organs (gills, digestive gland and foot) of M. galloprovincialis from representative Bulgarian Black Sea coastal habitats. The lipid peroxidation and glutathione levels, as well as activities of the antioxidant enzymes catalase, superoxide dismutase, glutathion peroxidase, glutathion reductase, glucose-6-phosphate dehydrogenase and glutathione-S-transferase of the organs were measured spectrophotometrically. Our hypothesis was that enhanced environmental pressure during the summer season, induced by multiple factors (biogenic, abiogenic and anthropogenic) led to weakening of the antioxidant protection in mussels at the beginning of autumn. The reaction of the mussel organism to the multiple stress factors was specific for the target organ and the type of the biomarker. Significant differences were present in the activity of the antioxidant system in mussels from the northern and southern coastal locations. The seasonal changes in the pro/antioxidant status of mussels were primarily due to specific seasonal changes in factors concerning the marine environment at the concrete locality. Further research is obviously needed to confirm the present results and provide a more complete data of seasonal and spatial changes in the antioxidant defense system of mussels from the Bulgarian Black Sea coastal area and their implementation in biomonitoring programs

State of the antioxidant defense system in wedge clams from Bulgarian Black Sea as a measure of resistance to environmental impacts

Pollution and climate change can induce oxidative stress (OS) in aquatic organisms. Reduced activity or incoordination between antioxidant enzymes in marine bivalves may cause cellular impairment with effects on higher levels of ecological organization. The present study aims to assess the condition factor and the activity of antioxidant enzymes catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione-S-transferase (GST), and glutathione concentration (GSH) in soft tissues of Donax trunculus (Linnaeus, 1758) as indicators of the state of marine ecosystems along the Bulgarian Black Sea coast. The wedge clams were sampled manually from different localities in June and in September. The activity of antioxidant enzymes and GSH concentrations varied both seasonally and among localities. Higher values of GSH, SOD and GPx were registered in wedge clams collected in autumn compared to those collected in summer. In wedge clams higher activity of the major phase II detoxification enzyme GST was observed in summer at localities with intensive tourism, suggesting an activation of the cell detoxification processes, presumably in response to increased environmental pressure. In conclusion, the observed presence of elevated enzyme activities suggested activation of the antioxidant protection system of the wedge clams in response to environmental pressure, indicating their ability to cope with induced OS and adapt to local conditions, and thus maintain ecosystem health

Comet assay - a sensitive tool for genotoxicity assessment of environmental stress in Mytilus galloprovincialis from the Bulgarian Black Sea coast

Assessment of DNA damage is of primary concern when the pollution-related stress in living organisms has to be determined. The reason for this concern comes from the evidence that damages in DNA often lead to mutations which are potential threat for the sustainability of organisms and ecological systems. To monitor genotoxicity of the marine environment we have developed a special procedure for application of the method of Comet assay (CA) on the native populations of Mytilus galloprovincialis. Samples were collected from areas with different anthropogenic load along the Bulgarian South Black Sea coast. As the first organ to encounter marine pollutants the gills of the mussels turned-out to be the most appropriate tissue for obtaining a single-cell suspension. Comet assay tests were performed and DNA damage was quantified using several different methods including percentage of DNA in the tail, Comet head and tail area, Comet number, Tail moment, Comet shape, etc. The method allowed us to monitor and predict the genetic risk of marine environmental stressors, particularly persistent pollutants. It proved to be convenient for precise quantification of the on-site loads of genotoxic stress on coastal ecosystems

Trace metal accumulation in tissues of wedge clams from sandy habitats of the Bulgarian Black Sea coast

The aim of the present study was to carry out an initial screening of trace metals bioaccumulation in soft tissues of the wedge clam (Donax trunculus Linnaeus, 1758) from different localities of the Bulgarian Black Sea coastal area and to evaluate the bioindicator potential of this species. Wedge clams were collected in June and September 2020 from sublittoral sandy habitats at different localities of the Bulgarian Black Sea coast. Soft tissues of individual clams were digested with nitric acid followed by ICP-OES analytical determination. The content of trace metals in the wedge clams differed significantly amongst localities. Higher metal content was present in wedge clams from Sveti Vlas, Shkorpilovtsi, Slanchev Bryag, Ahtopol and Kranevo. The highest values of lead (Pb) (2.51 mg/kg) and cadmium (Cd) (0.32 mg/kg) were found in samples from Sveti Vlas and the highest concentration of copper (Cu) (34.12 mg/kg), iron (Fe) (269.52 mg/kg) and nickel (Ni) (0.32 mg/kg) were detected in wedge clams from Shkorpilovtzi. Maximum content of chromium (Cr) (0.58 mg/kg) was present in samples from Slanchev Bryag, together with high values of Fe. The highest concentration of zinc (Zn) (18.04 mg/kg) together with high values of Cr and Fe were measured in wedge clams from Irakli. In conclusion, the wedge clams from the localities known to have higher coastal inflows and touristic pressures, i.e. Varna, Shkorpilovtzi, Sveti Vlas, Slanchev Bryag and Ahtopol accumulated significantly higher metal elements in their tissues. Only few significant seasonal differences in the concentration of metal elements in wedge clams were present and the observed seasonal variations were probably connected to the hydrological parameters of the ecosystems. The wedge clam D. trunculus is a suitable bioindicator for assessment and monitoring of metal pollution in the Bulgarian Black Sea environment

Comet assay - a sensitive tool for genotoxicity assessment of environmental stress in Mytilus galloprovincialis from the Bulgarian Black Sea coast

Assessment of DNA damage is of primary concern when the pollution-related stress in living organisms has to be determined. The reason for this concern comes from the evidence that damages in DNA often lead to mutations which are potential threat for the sustainability of organisms and ecological systems. To monitor genotoxicity of the marine environment we have developed a special procedure for application of the method of Comet assay (CA) on the native populations of Mytilus galloprovincialis. Samples were collected from areas with different anthropogenic load along the Bulgarian South Black Sea coast. As the first organ to encounter marine pollutants the gills of the mussels turned-out to be the most appropriate tissue for obtaining a single-cell suspension. Comet assay tests were performed and DNA damage was quantified using several different methods including percentage of DNA in the tail, Comet head and tail area, Comet number, Tail moment, Comet shape, etc. The method allowed us to monitor and predict the genetic risk of marine environmental stressors, particularly persistent pollutants. It proved to be convenient for precise quantification of the on-site loads of genotoxic stress on coastal ecosystems

Comparative study on the oxidative stress of commercially important fish species from localities with different ecological conditions along the Bulgarian Black Sea coast

The aim of the present study was to perform a pilot assessment and analysis of the oxidative stress (OS) level in four commercially important fish species (round goby, red mullet, sprat and horse mackerel) from different localities of the Bulgarian Black Sea coast. The fish were sampled during trawl selectivity experiments. The OS level in the fish was assessed by measuring lipid peroxidation (LPO), glutathione concentration (GSH), activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and glutathione-S-transferase (GST), as well as acetylcholine esterase (AChE) in gills and liver. Round goby and red mullet caught in the Nessebar Bay showed clear signs of OS with the highest levels of LPO and GST activities, accompanied by the lowest AChE activities in both liver and gills. On the contrary, round goby caught near Maslen Nos (a region with good ecological conditions) were least affected by OS with low LPO and high GSH concentrations and SOD activity. There were no significant differences in the OS bioindicators of horse mackerel from the different localities. Sprat caught in Nessebar Bay, compared to those caught from the other localities, showed presence of OS indicated by lower GSH levels and relatively higher CAT, GPx and GST activities, accompanied by low AChE activity in gills. It can be concluded that round goby and red mullet were more vulnerable to OS induced by marine environmental factors than the horse mackerel and sprat. However, their antioxidant defense system allows them to tolerate and adapt to the environment of their habitats. Further studies are needed for the assessment of OS in important fish species in the Bulgarian part of the Black Sea

A study of the microbiology of the intestinal tract in different species of Teleost fish from the Black Sea

This paper presents a study on the microbial status of different fish species and their habitats in the Bulgarian Black Sea area. The samples were collected in the period of January 2021 until March 2021. The fish species we used in this study were Black Sea turbot (Scophthalmus maximus), round goby (Neogobius melanostomus), shore rockling (Gaidropsarus mediterraneus) and European anchovy (Engraulis encrasicolus). The BIOLOG system was used for microbiological determination. From the different fish species, different species of microorganisms were isolated (using selective nutrient media). From the torbut, we isolated species Enterococcus villorum with 24 × 103 cells in 1 ml, Moraxella nonliquefaciens with 70 × 103 cells in 1 ml and Pseudomonas synxantha with 123 × 103 cells. Pseudomonas putida was isolated from the round goby with 20 × 103 cells in 1 ml. The species Streptococcus entericus with 123 × 103 cells in 1 ml was isolated from the shore rockling. Pseudomonas fulva with 60 × 103 cells in 1 ml was isolated from the European anchovy. A total of 223 × 103 cells in 1 ml of Pseudomonas agarici were isolated from Trachinus draco. Pseudomonas tolaasii with 145 × 103 cells in 1 ml were isolated from Merlangius merlangus. A different species of bacteria of the genus Pseudomonas was found for each of the investigated species of Black Sea fish. Apparently, the species Pseudomonas is characteristic of marine Teleostei and is important for the life and metabolism of these vertebrates. These microorganisms probably are resident species and developed not as result of pollution or environmental change