Impact of copper and zinc mixture on haematological parameters of rainbow trout (Oncorhynchus mykiss): acute exposure and recovery.

Significant changes in composition of rainbow trout Oncorhynchus mykiss blood cells types were induced after 4-days exposure with mixture of Cu2+ and Zn2+ at 0.25, 0.125 and 0.06 parts of LC50 in comparison to control group. The highest concentration of metal mixture (0.25 of LC50) significantly induced elevation of the number of monocytes and poly-segmented neutrophils. Treatment with 0.125 parts of LC50 concentration increased the number of thrombocytes, monocytes and non-segmented neutrophils. The most diluted mixture resulted in significant induction of thrombocytes, monocytes, non- and poly segmented neutrophils. Analysis of leucocyte cell types in the O. mykiss blood samples after 4-days of exposure at all applied mixture parts showed signs of monocytosis and neutrophilia. Comparison of different types of leucocytes' percentages (leukogram) in fish after 4-days exposure to metal mixture and after 4, 8, and 12-days recovery periods showed that, values of neutrophils even after the 12-days recovery period at all tested parts of LC50, and monocytes after exposure with the highest (0.25) used part of LC50 were not restored to control group levels. Depuration and recovery processes in treated fish are concentration and recovery period dependent.

The Impact of Intensive Fish Farming on Pond Sediment Microbiome and Antibiotic Resistance Gene Composition.

Aquaculture is a fast-growing animal food sector, and freshwater fish farming is particularly common in Central and Eastern Europe. As the biodiversity of fishery ponds is changed toward fulfilling the industrial needs, precautions should be taken to keep the system sustainable and protect the adjacent environment from possible damage. Due to risk of infectious diseases, antibiotics are used in aquaculture production systems. The constant exposure to antimicrobials can contribute to the rise of antibiotic resistance in aquaculture products and the adjacent ecosystems, with possibility of dissemination to the wider environment as well as between animals and humans. Even though previous studies have found antibiotic resistance genes in the sediments and water of farming ponds, the tendency and direction of spreading is not clear yet. The objective of this project was to evaluate the influence of intensive fish farming on the condition of water bodies used for the aquaculture and the environment, concentrating on the impact of the aquaculture on the surrounding water ecosystems as well as the possibility of transferring the pollutants and antibiotic resistance genes to both environment and the human hosts. Combined measurement of antibiotic and heavy metal contamination, toxicity assessment, microorganism diversity, and the detection of common antibiotic resistance genes was performed in the sediments of one fishery farm ponds as well as sampling points upstream and downstream. All the tested sediment samples did not show significantly elevated heavy metal concentrations and no substantial veterinary antibiotic pollution. From the antibiotic resistance genes tested, the presence of aminoglycoside and ß-lactam resistance determinants as well as the presence of integrons could be of concern for the possibility of transfer to humans. However, despite the lack of heavy metal and antibiotic pollution, the sediments showed toxicity, the cause of which should be explored more.

Genotoxic, cytotoxic, and neurotoxic responses in Anodonta cygnea after complex metal mixture treatment.

Environmental effects associated with the release of various metals even at maximum permissible concentrations (MPC) to the aquatic ecosystems are evident. In the present work, time-dependent increase in accumulated metals amount in gills of Anodonta cygnea after exposure to complex metal (Zn 0.1, Cu 0.01, Ni 0.01, Cr 0.01, Pb 0.005, and Cd 0.005 mg/L, MPC accepted for the inland waters in EU) mixture at various time points (1, 2, 4, 7, 14, and 28 days) was investigated. Statistically significant increase of Cu and Cd was determined in mussel's gills after 7-day exposure, in comparison to control group; moreover, significantly elevated concentration of Cu was measured and after 14-day treatment (in comparison to control and pre-exposure group). Concentrations of five (Cu, Ni, Cr, Pb, and Cd) out of 6 investigated metals were statistically increased in gills tissue after 28-day treatment. Moreover, complex metal mixture has demonstrated tissue- and time-dependent genotoxicity (∑Gentox) and cytotoxicity (∑Cytox) responses in mussels. After 4-day exposure, there were found the highest ∑Gentox levels in gills cells and haemocytes. Two-day treatment of mussels resulted in the highest and statistically significant induction of ∑Cytox level (in gills). Furthermore, after short-term (4 days) exposure, statistically significant inhibition of AChE activity in hemolymph of metal mixture-exposed mussels, in comparison to control and pre-exposure group, was found. Comparison of investigated responses in different tissue of A. cygnea discloses new information about metal mixture (at MPC) impacts at different treatment time. According to the obtained geno- and cytotoxicity data, it is suggested that gills are more sensitive tissue. Environmentally relevant trace metal concentrations when existing in mixture are able to cause adverse effects in A. cygnea; therefore, biological effects at different levels of organism are expected as a realistic scenario.

Environmental and synthetic sulphydryl group inhibitors: effects on bioluminescence and respiration in Vibrio fischeri.

Elemental sulphur (as S0 and S8) is abundant in anaerobic sediments and soil, and is highly toxic in the Vibrio fischeri bioluminescence test. This mode of S0 action remains uncertain. The objective of this research was the analysis of the toxic effects of S0 on bioluminescence and respiration in V. fischeri, in joint action with N-ethylmaleimide (NEM) or 2,4-dithio-DL-threitol (DTT), which are -SH group inhibiting and maintaining synthetic agents, respectively. Non-toxic DTT immediately protected cell bioluminescence against S0 inhibition at low (5.5ppb) and high (55ppb) concentrations of S0, whilst restoration of the inhibitory effect of S0 took up to 30 minutes. NEM (62.5ppb) diminished cell bioluminescence by up to 50% after 5 minutes, but after 60 minutes, the inhibition reached 100%. DTT restored the bioluminescence function inhibited in vivo and in vitro by S0 and NEM. Enhancement of cell respiration by up to 20% and 33% was observed at 2.2ppm of S0 and 36.8ppm of 2,4-dinitrophenol (2,4-DNP; an uncoupler of oxidative phosphorylation), respectively; whilst NEM (3.1ppm) caused a reduction of up to 40%. This comparative analysis confirmed that S0 has multiple modes of action--it acts as both an -SH group inhibitor and an uncoupler of oxidative phosphorylation in V. fischeri cells.