Combined evaluation of multi-biomarkers and metal bioaccumulations in two different fish species (Sparus aurata and Chelon labrosus) from Izmir Bay, Türkiye (Aegean Sea): Spatial, temporal and tissue-specific approaches.

Biomarkers were investigated to assess the effects of metal accumulation in Sparus aurata and Chelon labrosus in the Izmir Bay. Fish were collected from the Inner and Outer Bays in November 2020 and May 2021. According to the metal and biomarker measurements found in the organs, it was observed that the levels were higher in the liver and gill tissues than in the muscle tissues. Significant differences between tissues were found for all metals and biomarkers. In addition, biochemical biomarkers were found to be significant predictors of metal bioaccumulation. Histological changes were observed in liver and gills in each species and location. Although the metal levels determined according to the health risk assessment were below the threshold limits, the As levels for the lifetime cancer risk were within the limits to be considered. In conclusion, this study will provide robust results for its related study area.

Cellular Biomarkers in Mytilus galloprovincialis L. (Mediterranean Mussels) from Izmir Bay (Turkey).

In this study, cellular biomarkers and heavy metal concentration in the Mytilus galloprovincialis L. (Mediterranean mussels) collected from eight sites of Izmir Bay (Turkey) were determined to reveal water pollution for the first time in these stations. Results show that heavy metals (As, Cu, Hg, Zn, Cd, Sn, Pb) have been specified in mussels' tissues collected from all stations. According to GST, SOD, CAT activities, and TBARS contents, mussels in the outer bay have exposed more oxidative stress than the ones in the inner bay. Digestive gland tissues of them were showed more inhibition at AChE levels than gills. Also, abnormal nucleus rates and micronucleus frequencies (MN) were found to be higher in the inner bay than in the outer bay. This study showed that heavy metal pollution in different levels is an environmental issue on the Izmir Bay. Especially the coastal regions of the bay have been extremely affected by anthropogenic effects due to growing population.

How pollution effects the immune systems of invertebrate organisms (Mytilus galloprovincialis Lamark, 1819).

Hemocytes are the main immunologic parameters for invertebrate organisms as a response to diseases and pollutions. This study was aimed to reveal the relations between pollution and the total and differential hemocytes numbers. The gulf of Izmir was selected as location for sampling due to its contaminated by different anthropogenic pollutants mainly industrial effects. The effects of the pollutants were investigated on the mussels that have been collected from eight stations in the Izmir Bay (1- Inciralti, 2- Göztepe 3-Konak-4- Pasaport 5-Alsancak, 6-Karsiyaka, 7-Bostanli), which are known as the most polluted part of inner Bay of Izmir (Western Coast of Turkey) and 8-Foça, (an anthropogenically industrialized and shipping -impacted area). The pollution effects in the bay were determined on hemolymphs by biomarkers such as total hemocyte counts (THCs) and differential hemocyte counts (DHCs). Microscopic examinations have shown the statistical differences at THCs and DHCs. The highest total hemocyte number was determined in the station 7 (Bostanli) and the lowest in the station 4 (Pasaport) (P < 0.05). The number of total hemocyte counts were not differed significantly in the Stations among 2 (Göztepe), 5 (Alsancak) and 6 (Karsiyaka), and between Stations 8 (Foça) and 1 (Inciralti) (P > 0.05). In addition, there have been significant differences in subpopulation according to differential hemocyte numbers. The results indicate that the station 8 (reference site-Foça) has the highest agranulocyte number while station 7 (Bostanli) and the station 1 (Inciralti) have the lowest agranulocyte numbers (P < 0.05). As for the basophilic hemocyte numbers, this situation was reversed highest number in station 1 (Inciralti) and 7 (Bostanli) and lowest in the station 8 (Foça) (p < 0.05). The station 4 (Pasaport) has the highest eosinophilic hemocyte number while station 8 (Foça) has the lowest. We concluded that differential hemocytes counts are more sensitive as biomarker with the highest numbers of agranulocytes and lowest numbers of eosinophilic and basophilic hemocytes in reference site.

An investigation on the seasonal variations of the biomarkers of oxidative stress response and their correlations to Polonium-210 in mussel (Mytilus galloprovincialis) and common sole (Solea solea) from Izmir Bay, Turkey.

It is well known that the marine organisms are used as biological indicators for environmental pollution studies. Among these studies, the research on oxidative stress has been increasing in recent years. In this study, mussels (Mytilus galloprovincialis) and fish (Solea solea) samples were collected seasonally from Inciralti, Izmir, Turkey. This station was in an area where fishing is carried out for human consumption. The relationship between 210Po and oxidative stress markers (lipid peroxidation (LPO), H2O2 and proline) was investigated in the mussel tissue (digestive gland, gills) and fish tissue (liver, gills) samples. The present study indicated that H2O2 accumulated with increasing 210Po concentration in mussel samples. Statistically significant correlation were found between H2O2 and 210Po and LPO and proline in mussel samples. This correlation between LPO and proline can be attributed to common environmental parameters (other than 210Po) affecting expression of both LPO and proline levels. There was not a significant correlation between 210Po and LPO levels. Similarly, a significant correlation was not found between 210Po and proline.

Characterization of hemocytes from different body fluids of the eastern oyster Crassostrea virginica.

Bivalve hemocytes are involved in a variety of physiological and immunological functions. Circulating hemocytes in the hemolymph represent the main component of the internal self-defense system while hemocytes present in the extrapallial space (between the mantle and the shell) are actively involved in biomineralization and shell formation. This study focused on the characterization of hemocytes from different body fluids of the eastern oyster Crassostrea virginica. Hemocytes present in the hemolymph were compared to those contained in the extrapallial fluid. Hemocytes associated with the mucus layer covering pallial organs (mantle, gills, body wall) were also investigated because of their potential role as sentinel cells. Hemocytes were characterized using flow cytometry in conjunction with fluorescent epitope markers (clusters of differentiation, lectins) as well as functional assays (i.e. phagocytosis and reactive oxygen species -ROS). Compared with the hemolymph, there was a significantly greater percentage of granulocytes and agranulocytes among extrapallial and pallial hemocytes, respectively. Accounting for the different percentages of hemocyte sub-populations, significant differences in surface carbohydrate and clusters of differentiation signatures were also revealed between the different fluids. Most informative epitope markers included concanavalin A, peanut agglutinin, soybean agglutinin, CD11b and CD14. Functional assays revealed significant differences in phagocytic activity and ROS production between hemocytes from the extrapallial fluid and hemolymph; however, less robust differences were observed between hemolymph cells and hemocytes associated with the pallial mucus. Findings from this study suggest that there are markedly different hemocyte populations in the three body fluids. The role of peripheral cells, particularly those associated with the pallial mucus, requires further investigations.

Multi-Biomarker Responses After Exposure to Pollution in the Mediterranean Mussels (Mytilus galloprovincialis L.) in the Aegean Coast of Turkey.

In this study, sublethal effects on the Mediterranean mussels (Mytilus galloprovincialis L.) collected from the Aegean coast of Turkey were determined. Enzymes such as glutathione-S-transferase (GST), superoxide dismutase (SOD), catalase (CAT), and acetylcholinesterase (AChE), metallothionein (MT) mRNA expressions, thiobarbituric acid reactive substances (TBARS) contents, determination of 14 heavy metals and micronucleus frequency were selected as multibiomarkers. Results show that heavy metals and an increase in the level of MT gene expression have been determined in tissues of mussels collected from all stations. The GST, SOD and CAT enzymes were increased in mussels of Aliaga and Old Foca, compared to the mussels of Urla, while it was showed inhibition at AChE levels. Extensive LP is determined on mussels of Aliaga. It was determined that mussels in Aliaga region have exposed more oxidative stress than Old Foca and Urla. These biomarkers were carried out for the first time in these stations to assess environmental quality.

Histopathological and apoptotic changes on marine mussels Mytilus galloprovincialis (Lamark, 1819) following exposure to environmental pollutants.

Marine bivalve mussels, especially Mytilus species are an earlywarning system used for determining of damage caused by the various aquatic pollutions. In the present study, Mytilus galloprovincialis L. (black mussel) have been utilised as a biomonitoring organism to reveal environmental pollution in the Aliaga, Foca and Urla where located along the Izmir Coast of Turkey. Mussels were collected at these areas and gill and hepatopancreas (digestive gland) tissues were excised. mRNA expressions of initiator (caspase-2 and -8) and executioner (caspase -3/7-1, -3/7-2, -3/7-3 and -3/7-4) caspases of mussels tissues in areas exposed to pollution agent have been observed. TUNEL immunoreactivity in paralel to histopathological changes in both Aliaga and Foca areas were compared with Urla. This study is the first report to reveal the pollution with apoptotic expression on mussels in the coast of Turkey.

Assessment of micronuclei induction in peripheral blood and gill cells of some fish species from Aliaga Bay Turkey.

The aim of this paper was to assess the biological damages in fish caused by various mutagenic agents present in polluted waters of Aliaga Bay. For this purpose, micronuclei (MN) test was performed using peripheral erythrocytes and gill cells of different fish specimens caught from both polluted and relatively clean sites from Aliaga Bay (Turkey). Micronuclei tests is a system of mutagenicity testing used for determining changes in DNA fragments such as micronuclei in the cytoplasm of interphase cells caused by the pollution and chemicals in the environment. Thus, it was attempted to determine whether pollution affected the erythrocytes and gills of fish living in Aliaga Bay at the level of DNA by the means of micronuclei (MN) test. According to the results of present study, frequency of MN was found at high level in polluted site. In conclusion, this study indicates that the micronuclei test gives sensitive results in monitoring the pollution, especially the pollution of harbor, and thus it might be used as standard method in regularly monitoring pollution of coastal ecosystem.

Detecting micronuclei frequency in some aquatic organisms for monitoring pollution of Izmir Bay (Western Turkey).

Micronuclei tests is a system of mutagenicity testing used for determining the pollution and chemicals causing changes in DNA fragments such as micronuclei in the cytoplasm of interphase cells. Damage caused on the DNA by genotoxic pollutants is the first consequence occurring in the aquatic organisms. Thus, it was attempted to determine whether pollution affected the erythrocytes and gills of fish Gobius niger and haemolymph and gills of mussels Mytilus galloprovincialis living in Izmir Bay at the level of DNA by the means of micronuclei (MN) test. Organisms used in the MN test were collected from seven locations (Alsancak, Alaybey Shipyard, Karsiyaka, Bostanli, Göztepe, Konak and Pasaport) which are known as the most polluted part of inner Bay of Izmir (Western Coast of Turkey). According to the results of the present study, frequency of MN was found at high level in Alaybey Shipyard and Pasaport where wastes from existing dockyard contributed to high level of pollution. In conclusion, this study indicates that the micronuclei test gives sensitive results in monitoring the pollution, especially the pollution of harbor, and thus it might be used as standard method in regular monitoring of pollution of coastal ecosystem.