Post-exposure Recovery Research Articles

Characterizing transcriptomic responses of southern flounder (Paralichthys lethostigma) chronically exposed to Deepwater Horizon oiled sediments

To obtain a deeper understanding of the transcriptomic responses to oil in southern flounder (Paralichthys lethostigma), we performed quantitative PCR and RNA sequencing on liver and gill tissue after a chronic exposure (35 days) to Deepwater Horizon crude oiled sediment and after a 30-day recovery period. We wanted to understand which specific genes are differentially expressed in liver and gill tissues directly after oiled sediment exposure and with the addition of a recovery period. Furthermore, we wanted to examine specific enriched pathways in these two tissues to determine the impact of exposure with and without a recovery period on biological processes (e.g. immune function). Liver and gill tissues were chosen because they represent two distinct organs that are highly important to consider when examining the impacts of oiled sediment exposure. The liver is the classic detoxification organ, while the gill is in direct contact with sediment in benthic fishes. Examination of these two tissues, therefore, generates a broad understanding of the transcriptomic consequences of oil exposure across an organism. Gene expression for interleukin 8 (il8) and interleukin 1B (il1β) was significantly increased versus control measurements for fish exposed to oiled sediments for 35 days in gill tissue. Hierarchical clustering of gene expression showed that tissue type was the main driver of gene expression (rather than treatment). The inclusion of a 30-day post-exposure recovery period showed a return of il8 and il1β gene expression in the gill to baseline expression levels. However, the recovery period increased the number of differentially expressed genes and significantly affected canonical pathways in both tissue types. Pathways related to cholesterol biosynthesis were significantly suppressed in oil-exposed flounder with a recovery period, but not in the exposed flounder without a recovery period. At the end of the exposure, 17 pathways were significantly affected in the gill, including thyroid hormone metabolism-related pathways, which were the most influenced. Liver tissue from the recovered fish had the greatest number of enriched pathways for any tissue or time point (187). Cellular and humoral immune response pathways were considerably impacted in the liver after the recovery period, suggesting that the immune system was attempting to respond to potential damage caused from the chronic oil exposure. Our results demonstrate that liver and gill tissues from southern flounder were differentially altered by Deepwater Horizon oiled sediment exposure and that a 30-day recovery period after exposure substantially shifted gene expression and canonical pathway profiles.

A Multiple Life-History Trait-Based and Time-Resolved Assessment of Imidacloprid Effects and Recovery in the Widely Distributed Collembolan Folsomia quadrioculata.

Life-history traits determine individual fitness and the fate of populations. Imidacloprid, a widely used neonicotinoid insecticide, which persists in soil for more than 100 d at biologically relevant levels, may affect nontarget and ecologically important species, such as collembolans. In the present study, we determined the sublethal effects of short-term imidacloprid exposure and postexposure recovery in the collembolan Folsomia quadrioculata, which occurs abundantly across the northern hemisphere. We assessed survival, egg production, and hatching success in adult springtails exposed for 14 d through the diet to imidacloprid, followed by a 28-d postexposure phase. Survival and hatching success were high throughout the experiment in all the treatments, with no clear concentration dependence. However, egg production declined during the exposure phase and nearly stopped between 8 and 14 d in all the treatments (except the control) but resumed during the postexposure phase. Moreover, the resumption of egg production showed a concentration-dependent delay. Our findings suggest that low imidacloprid exposures can restrict reproduction, with potentially severe consequences for the population, notwithstanding the partial recovery in egg production. Environ Toxicol Chem 2021;40:139-147. © 2020 SETAC.

Effects of environmental cocaine concentrations on COX and caspase-3 activity, GRP-78, ALT, CRP and blood glucose levels in the liver and kidney of the European eel (Anguilla anguilla)

Cocaine is one of the most widely used illicit drugs in the world, and as a result of incomplete removal by sewage treatment plants it is found in surface waters, where it represents a new potential risk for aquatic organisms. In this study we evaluated the influence of environmental concentrations of cocaine on the liver and the kidney of the European eel (Anguilla anguilla). The eels were exposed to 20 ng L−1 of cocaine for fifty days, after which, three and ten days after the interruption of cocaine exposure their livers and kidneys were compared to controls. The general morphology of the two organs was evaluated, as well as the following parameters: cytochrome oxidase (COX) and caspase-3 activities, as markers of oxidative metabolism and apoptosis activation, respectively; glucose-regulated protein (GRP)78 levels, as a marker of endoplasmic reticulum (ER)-stress; blood glucose level, as stress marker; serum levels of alanine aminotransferase (ALT), as a marker of liver injury and serum levels of C-reactive protein (CRP), as a marker of the inflammatory process. The liver showed morphologic alterations such as necrotic areas, karyolysis and pyknotic nuclei, while the kidneys had dilated glomeruli and the renal tubules showed pyknotic nuclei and karyolysis. In the kidney, the alterations persisted after the interruption of cocaine exposure. In the liver, COX and caspase-3 activities increased (COX: P = 0.01; caspase-3: P = 0.032); ten days after the interruption of cocaine exposure, COX activity returned to control levels (P = 0.06) whereas caspase-3 activity decreased further (P = 0.012); GRP78 expression increased only in post-exposure recovery specimens (three days: P = 0.007 and ten days: P = 0.008 after the interruption of cocaine exposure, respectively). In the kidney, COX and caspase-3 activities increased (COX: P = 0.02; caspase-3: P = 0.019); after the interruption of cocaine exposure, COX activity remained high (three days: P = 0.02 and ten days: P = 0.029 after the interruption of cocaine exposure, respectively) whereas caspase-3 activity returned to control values (three days: P = 0.69 and ten days: P = 0.67 after the interruption of cocaine exposure, respectively). Blood glucose and serum ALT and CRP levels increased (blood glucose: P = 0.01; ALT: P = 0.001; CRP: 0.015) and remained high also ten days after the interruption of cocaine exposure (blood glucose: P = 0.009; ALT: P = 0.0031; CRP: 0.036). These results suggest that environmental cocaine concentrations adversely affected liver and kidney of this species.

How Reversible Are the Effects of Fumed Silica on Macrophages? A Proteomics-Informed View.

Synthetic amorphous silica is one of the most used nanomaterials, and numerous toxicological studies have studied its effects. Most of these studies have used an acute exposure mode to investigate the effects immediately after exposure. However, this exposure modality does not allow the investigation of the persistence of the effects, which is a crucial aspect of silica toxicology, as exemplified by crystalline silica. In this paper, we extended the investigations by studying not only the responses immediately after exposure but also after a 72 h post-exposure recovery phase. We used a pyrolytic silica as the test nanomaterial, as this variant of synthetic amorphous silica has been shown to induce a more persistent inflammation in vivo than precipitated silica. To investigate macrophage responses to pyrolytic silica, we used a combination of proteomics and targeted experiments, which allowed us to show that most of the cellular functions that were altered immediately after exposure to pyrolytic silica at a subtoxic dose, such as energy metabolism and cell morphology, returned to normal at the end of the recovery period. However, some alterations, such as the inflammatory responses and some aldehyde detoxification proteins, were persistent. At the proteomic level, other alterations, such as proteins implicated in the endosomal/lysosomal pathway, were also persistent but resulted in normal function, thus suggesting cellular adaptation.

Effects and recovery of larvae of the cold-water coral Lophelia pertusa (Desmophyllum pertusum) exposed to suspended bentonite, barite and drill cuttings

Fossil fuel drilling operations create sediment plumes and release waste materials into the ocean. These operations sometimes occur close to sensitive marine ecosystems, such as cold-water corals. While there have been several studies on the effects of energy industry activities on adult corals, there is very little information on potential impacts to their early life history stages. Larval stages of many marine organisms, including cold-water corals use cilia as a means of feeding and swimming, and if these structures become clogged with suspended particulates, the larvae may sink and be lost to the system.The objective of this study was to understand the response of Lophelia pertusa larvae to a different drilling waste components, and assess post-exposure recovery. Larvae of two ages (eight and 21 days) were exposed to a range of concentrations of bentonite, barite and drill cuttings. Larval sensitivity was assessed using the concentration at which 50% of the larvae showed behavioral effects (EC50) or lethal effects (LC50). Larvae showed greatest sensitivity to bentonite, followed by barite and drill cuttings, and also showed age-related responses that differed among the test materials. Post exposure recovery was variable across materials, with larvae exposed to bentonite having the lowest recovery rates. Understanding the vulnerability of early life history stages to human activities can help inform management strategies to preserve reproductive capacity of important marine ecosystems.

Neurotoxicity of perfluorooctanoic acid and post-exposure recovery due to blueberry anthocyanins in the planarians Dugesia japonica

Perfluorooctanoic acid (PFOA) is a widely used synthetic industrial chemical which accumulates in ecosystems and organisms. Our study have investigated the neurobehavioral effects of PFOA and the alleviation effects of PFOA-induced neurotoxicity by blueberry anthocyanins (ANT) in Dugesia japonica. The planarians were exposed to PFOA and ANT for ten days. Researchs showed that exposure to PFOA affected locomotor behavior and ANT significantly alleviated the reduction in locomotion induced by PFOA. The regeneration of eyespots and auricles was suppressed by PFOA and was promoted by ANT. Following exposure to PFOA, acetylcholinesterase activity continually decreased and was unaffected in the ANT group, but was elevated after combined administration of PFOA and ANT. Oxidative DNA damage was found in planarians exposed to PFOA and was attenuated after administration of ANT by the alkaline comet assay. Concentrations of three neurotransmitters increased following exposure to PFOA and decreased after administration of ANT. Furthermore, ANT promoted and PFOA inhibited neuronal regeneration. DjotxA, DjotxB, DjFoxG, DjFoxD and Djnlg associated with neural processes were up-regulated following exposure to PFOA. Our findings indicate that PFOA is a neurotoxicant while ANT can attenuate these detrimental effects.

Evaluation of protective effect of Thiocetam drug by morphological changes in the heart and vessels after administration of lead nanoparticles of various sizes (experimental study)

Introduction of nanotechnologies to the modern industry gave rise to new challenges. The issue of development and implementation of recommendations regarding the prevention of potential negative impact of lead nanoparticles on population health is of particular importance. The locally manufactured drug Thiocetam which possesses nootropic, antiischemic, antioxidative and membrane stabilizing properties has drawn our attention. The research aimed at studying the protective effect of Thiocetam in Wistar rats with simulated subchronic toxic effect of lead compounds of various sizes (by morphological changes in the heart and vessels). The experiments were conducted on Wistar rats (mean body weight of 160-180 g). Colloidal solutions of lead sulphide obtained by chemical synthesis with the use of sodium polyphosphate stabilizer, (PbS) with the mean size of 26-34 nm (1-PbS) and 50-80 nm (2-PbS), and lead nitrate Pb(NO3)2 (3-Pb) in the ionic form were used in simulating the toxic effect, while normal saline solution was administered to the control group. The studied substances were injected (intraperitoneally daily 5 times a week) in a dose of 0.94 mg/kg (in lead equivalent). The toxic effects were evaluated after 60 injections (three months) and one month after the discontinuation of exposure with and without Thiocetam. The drug Thiocetam in the dose of 250 mg/kg had been administered to rats intragastrically on an empty stomach daily for one month. Histological slides of the rats’ myocardium and aortal wall were studied and morphometric analysis and statistical processing performed. In the postexposure recovery period (PEP) period a lower degree of interstitial swelling and myocardial blood vessel filling was observed, which was considered to be a regression of damage. After the administration of Thiocetam a pronounced transverse striation of cardiomyocytes, the density of collagen fibers around cardiomyocytes and microvessels were revealed, which indicated the protective effect of pharmacological correction. However, leukocyte infiltration was also found in the myocardial or aortic microvessels in the experimental groups. Aortic morphometric data revealed no differences between the PbS NPs groups and Pb(NO3)2, although the aortic wall morphology was quite preserved. The use of Thiocetam prevented dystrophic changes in the atrial epicardium and the aortic adventitia, which indicate cytoprotective and connective tissue effects. In the postexposure period without pharmacological correction a tendency to spontaneous recovery of morphological changes of the heart and aortic walls under the influence of PbS NPs and lead nitrate was observed. However, morphometric parameters demonstrate the absence of complete recovery be it with or without Thiocetam.

Acute exposure to the water-soluble fraction of gasoline (WSFG) affects oxygen consumption, nitrogenous-waste and Mg excretion, and activates anaerobic metabolism in the goldfish Carassius auratus

Contamination of aquatic environments by petroleum and its products (e.g. gasoline) is a hazard for aquatic organisms as a result of the potential toxicity of monocyclic aromatic hydrocarbons (BTEX) and polycyclic aromatic hydrocarbons (PAH). Our goal was to evaluate the acute effects of the water-soluble fraction of gasoline (WSFG) on nitrogen excretion, osmoregulation, and metabolism of goldfish Carassius auratus. We first chemically characterized the WSFG and then tested its effects on these physiological aspects of C. auratus, in several different exposure scenarios (0, 0.25, 5, 10 and 25% of WSFG). The WSFG contained high concentrations BTEX (toluene 70% and benzene 17%) relative to PAH (<1%), and low levels of several metals (Al, Fe, Zn, Sr). Routine O2 uptake rate (MO2) of goldfish was inhibited by exposure to 5% WSFG, and during post-exposure recovery, MO2 increased in a dose-dependent fashion. Ammonia excretion was not affected by exposure to WSFG, but urea-N excretion increased progressively with the WSFG concentration. The same pattern of dose/response was observed for net Mg2+ loss rates and steadily increasing plasma lactate concentrations. Loss rates of Na+, Ca2+, K+ and Cl−, and plasma concentrations of Mg2+ and urea-N were not significantly altered. We propose that acute exposure to WSFG inhibits aerobic metabolism and activates anaerobic metabolism, breaking down ATP such that bound Mg2+ is liberated and the purine ring component is metabolized to urea-N, both of which are subsequently excreted.

Assessment the effect of exposure to microplastics in Nile Tilapia (Oreochromis niloticus) early juvenile: I. blood biomarkers

There is a scarcity of knowledge about the impacts of microplastics (MPs) on the early juvenile stage of freshwater fish. The current study aims to inspect the exposure and post-exposure recovery of microplastics (MPs) on accumulation and blood biomarkers of Nile Tilapia (Oreochromis niloticus) early juvenile. Four groups of fishes were used; the first group was the control group, the second group was exposed to (1 mg/L of MPs), the third group was exposed to (10 mg/L of MPs), and the fourth group was exposed to (100 mg/L of MPs) for 15 days and 15 days of recovery. The results showed that significantly higher numbers of microplastics were observed in microplastics-exposed groups compared to control group. Biochemical parameters (creatinine, uric acid, AST, ALT, ALP, glucose, cholesterol, total protein, albumin, globulin, and A/G ratio) showed significant increment after exposure to microplastics for 15 days compared to control group in dose dependent manner. The hematological indices (RBC's count, Hb, Ht, MCHC, Platelets, WBC's count, and monocytes) showed a significant decline after exposure to microplastics for 15 days compared to control group, while MCV and MCH showed a significant increase after exposure to microplastics for 15 days. After the recovery period, microplastics accumulations, hemato-biochemical alterations were still detected in microplastics exposed groups compared to the control group except for WBC's count and MCV which return to normal levels. MPs caused anemia and perturbations in hemato-biochemical parameters which may cause mortality of tilapia early juvenile and should be considered in a program for monitoring hazard materials in the ecosystem.

Amorphous-Carbon/Si Heterojunction Device for Room-Temperature NH3 Sensing

A room-temperature ammonia (NH <sub xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">3</sub> ) sensor based on amorphous-carbon/silicon(a-C/Si) heterojunction derived capacitive device is proposed in this article. Herein, controlled pyrolysis of lithographically patterned negative photoresist (SU8) thin films has been performed to realize highly stable a-C/Si junctions. Use of pyrolysis is an economic way of realizing the carbon films, as opposed to the conventional means of sputtering. Additionally, use of the photoresist allows us to preferably pattern the carbon film to develop heterojunction devices of varying shape and surface area. In this study, we have explored the NH <sub xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">3</sub> sensing properties of an unpatterned a-C film based heterojunction, where change in the capacitance-frequency (C-f) response of the heterojunction upon adsorption of NH <sub xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">3</sub> molecules on to the a-C surface is used as the transduction principle. The sensor accounts for a limit of detection (LoD) of 0.26 ppm, a response time of 5-10 s, a postexposure recovery time of 3-5 min, and excellent selectivity, in addition to facilitating room-temperature gas sensing.

Changes in the gills of the European eel (Anguilla anguilla) after chronic exposure to environmental cocaine concentration

The recent discovery of illicit drugs in the aquatic environment has raised concerns about the possible effects on the aquatic fauna, because of the pharmacological activity of these substances. Cocaine is an illicit drug widespread in surface waters since it is the third most widely used drug in North America, Western and Central Europe, and the second in Latin America and the Caribbean. The aim of this study was to evaluate the influence of environmental concentrations of cocaine on the gills of the European eel (Anguilla anguilla). The gills of male silver eels exposed to 20 ng L−1 of cocaine for fifty days were compared to control, vehicle control and post-exposure recovery ten days groups. The following parameters were evaluated: the thickness of the interlamellar epithelium (TIE), the length of the secondary lamellae (LSL) and the fraction of the interlamellar epithelium and the secondary lamellae occupied by the mucous cells (MC(IE-SL)FA) 3) the plasma cortisol and prolactin levels. After cocaine exposure, the gill epithelium appeared hyperplastic. The following changes were observed: proliferation in the interlamellar epithelium; partial and total fusion of the secondary lamellae, that appeared shortened and dilated; epithelial lifting and aneurism in the secondary lamellae. Moreover, in cocaine exposed eels, an increase in TIE and MC(IE-SL)FA and a decrease in LSL were observed. These changes were still present ten days after the interruption of cocaine exposure. Plasma levels of both cortisol and prolactin increased after cocaine exposure; ten days after the interruption of cocaine exposure, the plasma cortisol levels were still higher, whereas the plasma prolactin levels were lower, than control values. Our results show that even a chronic exposure to low environmental cocaine concentrations severely harms the eel gills, suggesting damages to their functions, and potentially affecting the survival of this species.

Effects of microplastics and mercury in the freshwater bivalve Corbicula fluminea (Müller, 1774): Filtration rate, biochemical biomarkers and mercury bioconcentration

The main objectives of this study were to investigate the effects of a mixture of microplastics and mercury on Corbicula fluminea, the post-exposure recovery, and the potential of microplastics to influence the bioconcentration of mercury by this species. Bivalves were collected in the field and acclimated to laboratory conditions for 14 days. Then, a 14-day bioassay was carried out. Bivalves were exposed for 8 days to clean medium (control), microplastics (0.13 mg/L), mercury (30 µg/L) and to a mixture (same concentrations) of both substances. The post-exposure recovery was investigated through 6 additional days in clean medium. After 8 and 14 days, the following endpoints were analysed: the post-exposure filtration rate (FR); the activity of cholinesterase enzymes (ChE), NADP-dependent isocitrate dehydrogenase (IDH), octopine dehydrogenase, catalase, glutathione reductase, glutathione peroxidase and glutathione S-transferases (GST), and the levels of lipid peroxidation (LPO). After 8 days of exposure to mercury, the bioconcentration factors (BCF) were 55 in bivalves exposed to the metal alone and 25 in bivalves exposed to the mixture. Thus, microplastics reduced the bioconcentration of mercury by C. fluminea. Bivalves exposed to microplastics, mercury or to the mixture had significantly (p ≤ 0.05) decreased FR and increased LPO levels, indicating fitness reduction and lipid oxidative damage. In addition, bivalves exposed to microplastics alone had significant (p ≤ 0.05) reduction of adductor muscle ChE activity, indicating neurotoxicity. Moreover, bivalves exposed to mercury alone had significantly (p ≤ 0.05) inhibited IDH activity, suggesting alterations in cellular energy production. Antagonism between microplastics and mercury in FR, ChE activity, GST activity and LPO levels was found. Six days of post-exposure recovery in clean medium was not enough to totally reverse the toxic effects induced by the substances nor to eliminate completely the mercury from the bivalve’s body. These findings have implications to animal, ecosystem and human health.

Effects of environmental cocaine concentrations on the skeletal muscle of the European eel (Anguilla anguilla)

The presence of illicit drugs in the aquatic environment represents a new potential risk for aquatic organisms, due to their constant exposure to substances with strong pharmacological activity. Currently, little is known about the ecological effects of illicit drugs. The aim of this study was to evaluate the influence of environmental concentrations of cocaine, an illicit drug widespread in surface waters, on the skeletal muscle of the European eel (Anguilla anguilla). The skeletal muscle of silver eels exposed to 20 ng L−1 of cocaine for 50 days were compared to control, vehicle control and two post-exposure recovery groups (3 and 10 days after interruption of cocaine). The eels general health, the morphology of the skeletal muscle and several parameters indicative of the skeletal muscle physiology were evaluated, namely the muscle whole protein profile, marker of the expression levels of the main muscle proteins; cytochrome oxidase activity, markers of oxidative metabolism; caspase-3, marker of apoptosis activation; serum levels of creatine kinase, lactate dehydrogenase and aspartate aminotransferase, markers of skeletal muscle damages. Cocaine-exposed eels appeared hyperactive but they showed the same general health status as the other groups. In contrast, their skeletal muscle showed evidence of serious injury, including muscle breakdown and swelling, similar to that typical of rhabdomyolysis. These changes were still present 10 days after the interruption of cocaine exposure. In fact, with the exception of the expression levels of the main muscle proteins, which remained unchanged, all the other parameters examined showed alterations that persisted for at least 10 days after the interruption of cocaine exposure. This study shows that even low environmental concentrations of cocaine cause severe damage to the morphology and physiology of the skeletal muscle of the silver eel, confirming the harmful impact of cocaine in the environment that potentially affects the survival of this species.

Influence of Pulsed Electric Fields and Mitochondria-Cytoskeleton Interactions on Cell Respiration

Pulsed electric fields with microsecond pulse width (μsPEFs) are used clinically; namely, irreversible electroporation/Nanoknife is used for soft tissue tumor ablation. The μsPEF pulse parameters used in irreversible electroporation (0.5–1 kV/cm, 80–100 pulses, ∼100 μs each, 1 Hz frequency) may cause an internal field to develop within the cell because of the disruption of the outer cell membrane and subsequent penetration of the electric field. An internal field may disrupt voltage-sensitive mitochondria, although the research literature has been relatively unclear regarding whether such disruptions occur with μsPEFs. This investigation reports the influence of clinically used μsPEF parameters on mitochondrial respiration in live cells. Using a high-throughput Agilent Seahorse machine, it was observed that μsPEF exposure comprising 80 pulses with amplitudes of 600 or 700 V/cm did not alter mitochondrial respiration in 4T1 cells measured after overnight postexposure recovery. To record alterations in mitochondrial function immediately after μsPEF exposure, high-resolution respirometry was used to measure the electron transport chain state via responses to glutamate-malate and ADP and mitochondrial membrane potential via response to carbonyl cyanide-p-trifluoromethoxyphenylhydrazone. In addition to measuring immediate mitochondrial responses to μsPEF exposure, measurements were also made on cells permeabilized using digitonin and those with compromised cytoskeleton due to actin depolymerization via treatment with the drug latrunculin B. The former treatment was used as a control to tease out the effects of plasma membrane permeabilization, whereas the latter was used to investigate indirect effects on the mitochondria that may occur if μsPEFs impact the cytoskeleton on which the mitochondria are anchored. Based on the results, it was concluded that within the pulse parameters tested, μsPEFs alone do not hinder mitochondrial physiology but can be used to impact the mitochondria upon compromising the actin. Mitochondrial susceptibility to μsPEF after actin depolymerization provides, to our knowledge, a novel avenue for cancer therapeutics.

Toxicity of mercury and post-exposure recovery in Corbicula fluminea: Neurotoxicity, oxidative stress and oxygen consumption

The toxicity of mercury to the invasive species Corbicula fluminea and the post-exposure recovery were investigated in relation to previous developmental exposure to distinct environmental conditions. Bivalves were collected in the estuaries of Minho River (M-est) and Lima River (L-est) that have several abiotic differences, including in environmental contamination, with the former being generally less contaminated. After 14 days of acclimation to laboratory conditions, two 14-day bioassays were performed simultaneously: one with bivalves from the M-est and the other with bivalves from the L-est. In each bioassay, the treatments were: dechlorinated tap water (clean medium) for 8 days, clean medium for 14 days, 31 µg/L of mercury for 8 days, 31 µg/L of mercury for 14 days and 31 µg/L of mercury for 8 days followed by 6 days in clean medium (post-exposure recovery). The effect criteria were the oxygen consumption rate (OCR), the activity of the enzymes cholinesterases (ChE), NADP-dependent isocitrate dehydrogenase (IDH), octopine dehydrogenase (ODH), catalase (CAT), glutathione reductase (GR), glutathione peroxidase (GPx) and glutathione S-transferases (GST) and lipid peroxidation (LPO) levels. Exposure to mercury for 8 days caused significant (p ≤ 0.05) inhibition of GR activity in M-est bivalves, whereas no significant adverse effects were observed in L-est animals. Moreover, evidences of delayed toxicity caused by 8-day exposure to mercury in OCR, IDH activity and LPO levels were found in M-est individuals but not in those of the L-est. Exposure to mercury for 14 days caused significant (p ≤ 0.05) depression of the OCR and of IDH activity in animals from both estuaries, indicating reduced individual fitness and hypoxia conditions. Moreover, oxidative stress and lipid peroxidation damage were observed in bivalves from the M-est exposed to mercury for 14 days but not in L-est animals. Differences in M-est and L-est environmental conditions to which animals were exposed in the wild likely contributed to the differences of sensitivity to mercury between M-est and L-est bivalves. The results of this study highlight the importance of investigating delayed toxicity, post-exposure recovery, and of taking into consideration the background contamination and other abiotic conditions of the original habitats when assessing the effects of environmental contaminants on animals from wild populations.

Transgenerational effects and recovery of microplastics exposure in model populations of the freshwater cladoceran Daphnia magna Straus

The environmental contamination by microplastics is a global challenge to ecosystem and human health, and the knowledge on the long-term effects of such particles is limited. Thus, the effects of microplastics and post-exposure recovery were investigated over 4 generations (F0, F1, F2, F3) using Daphnia magna as model. Effect criteria were parental mortality, growth, several reproductive parameters, and population growth rate. Microplastics exposure (0.1mg/l of pristine polymer microspheres 1–5μm diameter) caused parental mortality (10–100%), and significantly (p≤0.05) decreased growth, reproduction, and population growth rate leading to the extinction of the microplastics-exposed model population in the F1 generation. Females descending from those exposed to microplastics in F0 and exposed to clean medium presented some recovery but up to the F3 generation they still had significantly (p≤0.05) reduced growth, reproduction, and population growth rate. Overall, these results indicate that D. magna recovery from chronic exposure to microplastics may take several generations, and that the continuous exposure over generations to microplastics may cause population extinction. These findings have implications to aquatic ecosystem functioning and services, and raise concern on the long-term animal and human exposure to microplastics through diverse routes.

Acute effects of ammonia exposure on the plasma and haematological parameters and histological structure of the juvenile blunt snout bream, Megalobrama amblycephala , and post-exposure recovery

In this study, we investigated the toxic effects of ammonia-N on the plasma and haematological parameters and histological structure of blunt snout bream (Megalobrama amblycephala) juveniles. The fish (initial weight, 14.79 ± 0.01 g) were randomly sorted into six tanks (200 L), and each tank was stocked with 40 fish for culture. The juveniles were exposed to two ammonia-N levels—0 mg/L (control group) and 25 mg/L (experimental group)—and sampled at 0, 6, 12, 24, 48 and 72 hr, and then they underwent 96 hr of post-exposure recovery. The results showed that ammonia-N had significant effects on the plasma and haematological parameters. The treatment group showed increased cortisol, plasma ammonia and haematocrit levels and white blood cell count with increasing exposure time, up to 24 hr, and then the levels and count decreased. A significantly higher plasma glucose level was observed in the treatment group at 12 hr. After 96 hr of post-exposure recovery, all parameter levels decreased to the control levels. The fish displayed histopathological alterations in the gills, liver and kidney. The results indicate that the severity of the lesions clearly differed among the organs, with the liver showing the most extensive damage, followed by the gills and kidney. Adverse effects to physiological indicators and histological structure increased with increasing exposure time before 24 hr. The fish showed self-regulation; however, the histological structure could not recover fully, the gill tissue showed irreversible changes and the kidney tissue exhibited the worst recovery ability.

Peroxiredoxin 6 phospholipid hydroperoxidase activity in the repair of peroxidized cell membranes

Although lipid peroxidation associated with oxidative stress can result in cellular death, sub-lethal lipid peroxidation can gradually resolve with return to the pre-exposure state. We have shown that resolution of lipid peroxidation is greatly delayed in lungs or cells that are null for peroxiredoxin 6 (Prdx6) and that both the phospholipase A2 and the GSH peroxidase activities of Prdx6 are required for a maximal rate of recovery. Like other peroxiredoxins, Prdx6 can reduce H2O2 and short chain hydroperoxides, but in addition can directly reduce phospholipid hydroperoxides. This study evaluated the relative role of these two different peroxidase activities of Prdx6 in the repair of peroxidized cell membranes. The His26 residue in Prdx6 is an important component of the binding site for phospholipids. Thus, we evaluated the lungs from H26A-Prdx6 expressing mice and generated H26A-Prdx6 expressing pulmonary microvascular endothelial cells (PMVEC) by lentiviral infection of Prdx6 null cells to compare with wild type in the repair of lipid peroxidation. Isolated lungs and PMVEC were exposed to tert-butyl hydroperoxide and mice were exposed to hyperoxia (> 95% O2). Assays for lipid peroxidation in wild type control and mutant lungs and cells showed ~4-fold increase at end-exposure. Control lungs and cells showed gradual resolution during a post-exposure recovery period. However, there was no recovery from lipid peroxidation by H26A-Prdx6 lungs or PMVEC. These studies confirm an important role for Prdx6 in recovery from membrane lipid peroxidation and indicate that reduction of H2O2 or short chain hydroperoxides does not play a role in the recovery process.

Comparative toxicological effects of two antifouling biocides on the marine diatom Chaetoceros lorenzianus: Damage and post-exposure recovery

Antifouling biocides are commonly used in coastal electric power stations to prevent biofouling in their condenser cooling systems. However, the environmental impact of the chemical biocides is less understood than the thermal stress effects caused by the condenser effluents. In this study, Chaetoceros lorenzianus, a representative marine diatom, was used to analyse the toxicity of two antifouling biocides, chlorine and chlorine dioxide. The diatom cells were subjected to a range of concentrations of the biocides (from 0.05 to 2mg/L, as total residual oxidants, TRO) for contact time of 30min. They were analysed for viability, genotoxicity, chlorophyll a and cell density endpoints. The cells were affected at all concentrations of the biocides (0.05–2mg/L), showing dose-dependent decrease in viability and increase in DNA damage. The treated cells were later incubated in filtered seawater devoid of biocide to check for recovery. The cells were able to recover in terms of overall viability and DNA damage, when they had been initially treated with low concentrations of the biocides (0.5mg/L of Cl2 or 0.2mg/L of ClO2). Chlorophyll a analysis showed irreparable damage at all concentrations, while cell density showed increasing trend of reduction, if treated above 0.5mg/L of Cl2 and 0.2mg/L of ClO2. The data indicated that in C. lorenzianus, cumulative toxic effects and recovery potential of ClO2 up to 0.2mg/L were comparable with those of Cl2, up to 0.5mg/L concentration in terms of the studied endpoints. The results indicate that at the biocide levels currently being used at power stations, recovery of the organism is feasible upon return to ambient environment. Similar studies should be carried out on other planktonic and benthic organisms, which will be helpful in the formulation of future guidelines for discharge of upcoming antifouling biocides such as chlorine dioxide.

Static Magnetic Field Effect on Cell Alignment, Growth, and Differentiation in Human Cord-Derived Mesenchymal Stem Cells.

This investigation is performed to evaluate the impact of static magnetic field on the Cell growth alignment, and differentiation potential in Human Mesenchymal Stem cells derived from human newborn cords. In vitro-cultured mesenchymal stem cells derived from human newborn cords were exposed to SMF up to 24mT and compared with the control (unexposed) cultures. Viability was assessed via Trypan Blue staining and MTT assay. Cell cycle progression was studied after flow cytometry data analysis. Sox-2, Nanong, and Oct-4 Primers used for RT-PCR experiment. Morphological studies showed that the exposed cells were significantly aligned in parallel bundles in a correlation with the magnetic field lines. Viability measurements showed a significant reduction in cell viability which was noted after exposure to static magnetic field and initiated 36h after the end of exposure time. Flow cytometric data analysis confirmed a decrease in G1 phase cell population within the treated and cultured groups compared with the corresponding control samples. However, the induced changes were recovered in the cell cultures after the post-exposure culture recovery time which may be attributed to the cellular repair mechanisms. Furthermore, the proliferation rate and Oct-4 gene expression were reduced due to the 18mT static magnetic field exposure. The significant proliferation rate decrease accompanied by the Sox-2, Nanong, and Oct-4 gene expression decline, suggested the differentiation inducing effects of SMF exposure. Exposure to Static Magnetic fields up to 24mT affects mesenchymal stem cell alignment and proliferation rate as well as mRNA expression of Sox-2, Nanong, and Oct-4 genes, therefore can be considered as a new differentiation inducer in addition to the other stimulators.