Lysosomal Membrane Stability of Mussel (Mytilus galloprovincialis Lamarck, 1819) as a Biomarker of Cellular Stress for Environmental Contamination.

The lysosomal membrane stability (LMS) of hemocytes in wild mussels (Mytilus galloprovincialis) as a biomarker of cellular stress for chemical pollution was tested by neutral red retention time (NRRT) assays. To assess the environmental contamination in the study area, seawater quality and pollutant bioaccumulation throughout the soft tissue of mussels were investigated. The samples were collected in July 2022 at four sites on the Romanian Black Sea coast considered to be differently affected by contamination. To support the suitability of LMS as a biomarker of contaminant-induced stress, the contaminant body burdens of the mussels were compared with the NRRT values. The results showed a significantly reduced NRRT in all investigated locations, particularly in port areas (mean retention time between 11 and 14 min). The elevated bioaccumulation of organochlorinated pesticides (OCPs) and polychlorinated biphenyls (PCBs) and low NRRTs were observed at the most contaminated sites (i.e., ports). The low lysosomal stability reflected stress and damage in the hemocytes of mussels and could be related to the body burdens of contaminants. LMS is an effective indicator of health status in mussels and could be considered a sensitive biomarker of cellular stress induced by contaminant exposure.

Multiple Assays on Non-Target Organisms to Determine the Risk of Acute Environmental Toxicity in Tebuconazole-Based Fungicides Widely Used in the Black Sea Coastal Area.

The widespread use of Tebuconazole-based fungicides in phytosanitary treatments on a wide range of crops, on the one hand, and the lack of official reports on the amount of fungicide residues in nearby water basins, on the other hand, may lead to uncontrolled and hazardous contamination of water sources used by the resident population, and to serious effects on the environment and public health. Our study explores the acute toxicological risk of this fungicide on various organisms, from bacteria and yeast to fish, using a battery of tests (standardized Toxkit microbiotests and acute semi-static tests). By investigating the interaction between Tebuconazole and bacteria and yeast organisms, we observed that Gram-negative bacteria displayed a strong tolerance for Tebuconazole, while Gram-positive bacteria and yeasts proved to be very sensitive. The fish experiment was conducted on Chelon auratus juveniles exposed to five concentrations of the fungicide Tebustar EW (Tebuconazole, 250 g/L as active substance). After 96 h of exposure, the LC50 for C. auratus was 1.13 mg/L. In the case of the Toxkit microbiotests' application, the following results were recorded: Spirodela polyrhiza EC50 = 2.204 mg/L (after 72 h exposure), Thamnocephalus platyurus EC50 = 0.115 mg/L (after 24 h), and Daphnia magna EC50 = 2.37 mg/L (after 24-48 h). With the exception of bacteria and yeast, the same response pattern was observed for all non-target species tested; the response range expressed by concentrations causing growth inhibition or mortality was small, ranging between very close values that are quite low, thereby demonstrating the high toxicity of Tebuconazole-based fungicides to the environment.

Correlation Between Toxic Elements and Pesticide Residues in Medicinal Herbs Available in Pharmaceutical Market.

The use of medicinal plants for self-medication of minor health conditions has become a widespread practice in contemporary society. Few consumes, however, question the contamination of these products with toxic factors resulting from the planet's increasingly polluted environment. This paper presents the levels of five toxic elements (As, Cr, Pb, Cd, and Hg) and nine organochlorine pesticides (hexachlorobenzene (HCB), lindane, heptachor, aldrin, dieldrin, endrin, p,p'DDE, p,p'DDD, and p,p'DDT) in 14 brands of regularly consumed medicinal products in Romania. The toxic elements content was determined using energy-dispersive X-ray fluorescence (EDXRF) technique, and organochlorine pesticide residues (OPCs) were quantified using gas-chromatographic method, equipped with electron capture detector (GC-ECD). The results show that in the case of Cr, Cd, and Hg, the concentrations exceeded the limit values established by World Health Organisation (WHO) for raw herbal material. The higher level of OPCs (such as p,p'DDD, p,p'DDT, aldrin, and dieldrin) was found in the samples of Hypericum perforatum-St. John's wort, Crataegus monogyna-hawthorn, and Epilobium parviflorum-hoary willowherb. The correlations between the content of toxic elements and pesticides were determined by statistical analysis. Hierarchical clustering technique was used to detect natural grouping between the toxic elements and pesticides. For herb samples, four clusters were identified, the strongest correlated cluster consisting of Pb, HCB, Cr, and Hg. A further analysis within this cluster suggested that Cr levels are statistically different from the rest of the elements.

Suitability and Sensitivity of Golden Grey Mullet Chelon auratus (Risso, 1810) as a Reference Fish Species for Ecotoxicity Tests in the Black Sea.

In recent years, hydrocarbon exploration and production operations have intensified in the Black Sea. Alongside growth in exploration and production activities, the influence of chemical usage across multiple industrial sectors within the Black Sea environment has become increasingly interesting. The aim of this research was to define a protocol for determining the acute toxicity of chemicals using the golden grey mullet, Chelon auratus (Risso, 1810), a native pelagic fish species of the Black Sea. Juvenile golden grey mullets were exposed for 96 h, under semi-static conditions, to dilutions of the reference toxicant 3,5-Dichlorophenol. Results from three reference toxicity tests (LC50 = 1.25 mg/L, 1.739 mg/L, and 1.409 mg/L) indicated that C. auratus is of moderate sensitivity when compared to literature values from EPAs Ecotox database. The protocol described within is intended to ensure Black Sea native organisms are represented by standard hazard assessment practices.

Polycyclic aromatic hydrocarbons and pesticides in milk powder.

This Research Communication reports analysis of 37 compounds comprising polycyclic aromatic hydrocarbons (PAHs), organochlorine and organophosphate pesticides (OCPS and OPPS) in milk powder (one brand each of commercial infant formulae, follow-on formulae and baby formulae purchased from a local supermarket in Romania). The selected analytes were investigated using gas chromatography-mass spectrometry (GC-MS), gas chromatography with electron capture detector (GC-ECD) and gas chromatography with thermionic sensitive detection (GC-TSD). The estimated limits of detection for most target analytes were in the µg/kg level (range 0·001-0·320 µg/kg). The purpose of the study was to determine the selected analytes, to assess the exposure of babies and infants and to produce data for comparison with tolerable limits according to the European Union Regulations. In most of the samples the organochlorine pesticides values were under the limit of detection. Exceptions were heptachlor epoxide and endosulfan sulphate, the last of which was found in all analysed samples at low concentrations. We also found detectable levels of ethoprophos, parathion-methyl, chlorpyrifos, prothiofos, guthion, disulfoton and fenchlorphos in most of the analysed samples. Benzo[a]pyrene, which is used as an indicator for the presence of PAHs, was not detected in selected samples. The low level of exposure to contaminants indicates that there are no health risks for the infants and babies that consume this brand of milk powder formulae.

Biochemical biomarker responses to pollution in selected sentinel organisms across the Eastern Mediterranean and the Black Sea.

Pollution effects were assessed by means of biochemical biomarkers (catalase, glutathione S-transferase and acetylcholinesterase activities, and metallothioneins content) in five species at selected coastal sites across the Eastern Mediterranean and the Black Sea. The mussel Mytilus galloprovincialis, a well-established sentinel species, was investigated in the Adriatic Sea, Aegean Sea, and Black Sea. The mussel Brachidontes pharaonis and the striped red mullet Mullus surmuletus were used in the Levantine Sea where M. galloprovincialis is not present. The white seabream Diplodus sargus sargus and the gastropod Rapana venosa were additionally sampled in the Adriatic and the Black Sea, respectively. Mussels showed catalase, glutathione S-transferase, and acetylcholinesterase responses to pollution in most geographical areas while the response of metallothioneins was restricted to a few sites. R. venosa showed marked responses of catalase and metallothioneins whereas both fish species did not generally exhibit variations in biomarker values among sites. The approach based on the reference deviation concept using the "Integrated Biological Responses version 2" index was useful for the interpretation of overall biomarker responses.