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Cocaine (COC) is a powerful illicit drug frequently detected in the aquatic environment. COC acts by inhibiting the reuptake of dopamine (DOPA) and 5-hydroxytryptamine (5-HT - serotonin) and causes endocrine disturbances in mammals. This study investigated similar effects from cocaine exposure in the marine mussel Perna perna, as well as neurotoxicity and energy imbalances. Mussels were exposed to COC (0.2 μg.L -1 and 2 μg.L -1 ) for periods of 48, 96, and 168 h. Acetylcholinesterase (AChE) was measured in adductor muscle tissue to determine neurotoxicity, and neurotransmitter levels (DOPA and 5-HT), monoamine oxidase (MAO) and cyclooxygenase (COX) activity, and energy status (mitrochondrial electron transport, MET, and total lipids, TLP) were evaluated in the mussels' gonads. COC decreased AChE activity in the mussels exposed to 0.2 μg.L -1 and 2 μg.L -1 after 168 h, and all concentrations of COC increased neurotransmitter levels. Increases in MET (0.2 μg.L -1, for all exposure periods) and TLP (0.2 μg.L 1 after 48 h, and 2 μg.L -1 after 96 h and 168 h) were also observed. No significant change was detected in MAO activity. COC also decreased COX activity in the mussels exposed to 0.2 μg.L -1 (48 h and 96 h) and 2 μg.L -1 (96 h). These results suggest that COC may compromise neurotransmitter levels and COX activity. Furthermore, the changes in MET and LPT suggest that COC affects the energy balance of the mussels, and could negatively affect physiological processes such as metabolism, hormone production, and embryonic development., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Offshore oil exploration creates threats to coastal ecosystems, including increasing urbanization and associated effluent releases. Genotoxicity biomarkers in mussels were determined across a gradient of coastal zone influences of offshore petroleum exploration in southeastern Brazil. Coastal ecosystems such as estuaries, beaches and islands were seasonally monitored for genotoxicity evaluation using the brown mussel Perna perna. The greatest DNA damage (5.2% ± 1.9% tail DNA and 1.5‰  ± 0.8‰ MN) were observed in urban estuaries, while Santana Archipelago showed levels of genotoxicity near zero and is considered a reference site. Mussels from urban and pristine beaches showed intermediate damage levels, but were also influenced by urbanization. Thus, mussel genotoxicity biomarkers greatly indicated the proposed oil exploration and urbanization scenarios that consequently are genetically affecting coastal organisms.

The present study investigated the toxicity of several emerging contaminants: the pharmaceutical drug carbamazepine (CBZ), the plasticizer bisphenol A (BPA), and the herbicide atrazine (ATZ) in a marine bivalve. Green mussels (Perna viridis) were exposed to different concentrations of CBZ, BPA, and ATZ, either individually or as mixtures over a 7-d period, and a suite of molecular and cellular biomarkers were analyzed: biomarkers of immunotoxicity (total hemocyte count, phagocytosis, extracellular lysozyme), genotoxicity (Comet assay), neurotoxicity (inhibition of acetylcholinesterase [AChE]), endocrine disruption (vitellin-like proteins), and detoxification enzymes (cytochrome P4501A [CYP1A], 7-ethoxyresorufin O-deethylase [EROD], and glutathione-S-transferase [GST]). Results of the single-chemical exposure tests highlighted the relatively low toxicity of CBZ because most biomarker responses observed were recorded at concentrations well above environmental levels. Bisphenol A exposure at environmentally realistic concentrations resulted in clear immunomodulatory, genotoxic, and endocrine-disruptive effects. Similarly, 3 of the 10 biomarkers tested on green mussels (genotoxicity, inhibition of AchE, and EROD) responded after exposure to ATZ at environmentally relevant doses or below, and confirmed the potency of this herbicide to marine bivalves. Exposure tests using mixtures of CBZ, BPA, and ATZ also revealed that these 3 substances were generally acting in an additive manner on the selected biomarkers, at environmental doses, with some exceptions (antagonism and/or synergy) at low and high concentrations. The present study also confirms that most of the biomarkers used are suitable for biomonitoring studies with green mussels. Environ Toxicol Chem 2017;36:429-441. © 2016 SETAC., (© 2016 SETAC.)

The green mussel Perna viridis is an ideal biomonitor to evaluate marine environmental pollution. Benzo(a)pyrene (BaP) is a typical polycyclic aromatic hydrocarbon (PAH), which is well known for the mutagenic and carcinogenic characteristics. However, the toxicological effects of BaP on Perna viridis embryo are still unclear. In this study, we investigated the embryo transcriptomic profile of Perna viridis treated with BaP via digital gene expression analysis. A total of 92,362,742 reads were produced from two groups (control and BaP exposure) by whole transcriptome sequencing (RNA-Seq). Gene Ontology (GO) and Kyoto Encyclopaedia of Genes and Genomes (KEGG) pathway analysis were used on all genes to determine the biological functions and processes. Genes involved in various molecular pathways of toxicological effects were enriched further. The differential expression genes (DEGs) were related to stress response, infectious disease and innate immunity. Quantitative real-time PCR (qRT-PCR) measured expressional levels of six genes confirmed through the DGE analysis. This study reveals that RNA-seq for transcriptome profiling of P. viridis embryo can better understand the embryo toxic effects of BaP. Furthermore, it also suggests that RNA-seq is a superior tool for generating novel and valuable information for revealing the toxic effects caused by BaP at transcriptional level., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

This study aims to test if RNA/DNA ratio and various energy reserve parameters (i.e., glycogen, lipid, protein content and total energy reserves) are sensitive biomarkers for indicating stresses induced by metal contaminants in the green-lipped mussel Perna viridis, a common organism for biomonitoring in Southeast Asia. This study was, therefore, designed to examine the effects of cadmium (Cd) and copper (Cu) on these potential biomarkers in two major energy storage tissues, adductor muscle (AM) and hepatopancreas (HP), of P. viridis after sub-lethal exposure to either metal for 10 days. The results showed that neither Cd nor Cu treatments affected the RNA/DNA ratio, glycogen and protein contents in AM and HP. As the most sensitive biomarker in P. viridis, the total lipid content in both AM and HP was significantly decreased in the treatment of 5μg Cu/L and 0.01-0.1μgCd/L, respectively. However, soft-tissue body burdens of Cu or Cd did not significantly correlate with each of the four biomarkers regardless of the tissue type. In addition, AM generally stored more glycogen than HP, whereas HP stored more lipids than AM. We proposed that multiple biomarkers may be employed as an integrated diagnostic tool for monitoring the health condition of the mussels., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Benzo(a)pyrene (BaP) and dichlorodiphenyltrichloroethane (DDT) are persistent organic pollutants and environmental estrogens (EEs) with known toxicity towards the green mussel, Perna viridis. In this study, the toxic effects of BaP (10 µg/L) and DDT (10 µg/L) and their mixture were assessed in green mussel gills with proteomic and metabolomic approaches. Metabolic responses indicated that BaP mainly caused disturbance in osmotic regulation by significantly decrease in branched chain amino acids, dimethylamine and dimethylglycine in gills of male green mussels after exposure for 7 days. DDT mainly caused disturbance in osmotic regulation and energy metabolism by differential alteration of betaine, dimethylamine, dimethylglycine, amino acids, and succinate in gills of male green mussels. However, the mixture of BaP and DDT didn't show obvious metabolite changes. Proteomic analysis showed different protein expression profiles between different treatment groups, which demonstrated that BaP, DDT and their mixture may have different modes of action. Proteomic responses revealed that BaP induced cell apoptosis, disturbance in protein digestion and energy metabolism in gills of green mussels, whereas DDT exposure altered proteins that were associated with oxidative stress, cytoskeleton and cell structure, protein digestion and energy metabolism. However, the mixture of BaP and DDT affected proteins related to the oxidative stress, cytoskeleton and cell structure, protein biosynthesis and modification, energy metabolism, growth and apoptosis., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Perna viridis (P. viridis) has been identified as a good biological indicator in identifying environmental pollution, especially when there are various types of Heavy Metals Accumulations (HMA) inside its tissue. Based on the potential of P. viridis to accumulate heavy metals and the data on its physical properties, this study proffers to determine the relationships between both properties. The similarities of the physical properties are used to mathematical model their relationships, which included the size (length, width, height) and weight (wet and dry) of P. viridis, whilst the heavy metals are focused on concentrations of Pb, Cu, Cr, Cd and Zn. The concentrations of metal elements are detected by using Flame Atomic Adsorption Spectrometry. Results show that the mean concentration of Pb, Cu, Cr, Cd, Zn, length, width, height, wet weight and dry weight are: 1.12 +/- 1.00, 2.36 +/- 1.65, 2.12 +/- 2.74, 0.44 +/- 0.41 and 16.52 +/- 10.64 mg kg(-1) (dry weight), 105.08 +/- 14.35, 41.64 +/- 4.64, 28.75 +/- 3.92 mm, 14.56 +/- 3.30 and 2.37 +/- 0.86 g, respectively. It is also found out that the relationships between the Heavy Metals Concentrations (HMA) and the physical properties can be represented using Multiple Linear Regressions (MLR) models, relating that the HMA of Zinc has affected significantly the physical growth properties of P. viridis.

Distribution of heavy metals and its associated histological perturbations were studied in the soft tissues of Perna viridis collected from Ennore estuary and compared with the less polluted Kovalam coast. The concentration of copper, lead, zinc, cadmium, manganese and iron were quantified in gills, digestive gland and adductor muscle. The results showed marked differences between the two sites as well as significant variations within the tissues. Among the heavy metals analyzed, lead and cadmium recorded very low in the soft tissues of mussel. Heavy metal levels in tissues of mussel collected from Ennore estuary were in the order of gills>digestive gland>adductor muscle, while it was digestive gland>gills>adductor muscle in the mussel sampled from Kovalam coast. The decreasing trend of metals in the tissues of mussels sampled from both Ennore estuary and Kovalam coast was in the order of Fe>Mn>Zn>Cu>Pb>Cd. Overall, the highest metal concentrations were found in the mussel collected from Ennore estuary. The metal accumulation in the gills and digestive gland of Perna viridis was found to be quite high in comparison with the adductor muscle. These soft tissues were further investigated by light microscopy and the results were compared with the reference site (Kovalam coast). These results suggest that thickening of the digestive epithelium, hemocytic infiltration in the gills and myodegeneration in the muscle tissue are useful histological biomarkers for heavy metal induced stress, and demonstrate that precautions need to be taken in Ennore estuary in order to prevent heavy metal pollution that can occur in the future., (Copyright © 2012 Elsevier Inc. All rights reserved.)

This study aimed to provide the first biomonitoring integrating biomarkers and bioaccumulation data in São Paulo coast, Brazil and, for this purpose, a battery of biomarkers of defense mechanisms was analyzed and linked to contaminants' body burden in a weigh-of-evidence approach. The brown mussel Perna perna was selected to be transplanted from a farming area (Caraguatatuba) to four possibly polluted sites: Engenho D'Água, DTCS (Dutos e Terminais do Centro-Oeste de São Paulo) oil terminal (Sao Sebastiao zone), Palmas Island, and Itaipu (It; Santos Bay zone). After 3 months of exposure in each season, mussels were recollected and the cytochrome P4501A (CYP1A)- and CYP3A-like activities, glutathione-S-transferase and antioxidants enzymes (catalase, glutathione peroxidase, and glutathione reductase) were analyzed in gills. The concentrations of polycyclic aromatic hydrocarbons, linear alkylbenzenes, and nonessential metals (Cr, Cd, Pb, and Hg) in whole tissue were also analyzed and data were linked to biomarkers' responses by multivariate analysis (principal component analysis-factor analysis). A representation of estimated factor scores was performed to confirm the factor descriptions and to characterize the studied stations. Biomarkers exhibited most significant alterations all year long in mussels transplanted to It, located at Santos Bay zone, where bioaccumulation of organic and inorganic compounds was detected. This integrated approach using transplanted mussels showed satisfactory results, pointing out differences between sites, seasons, and critical areas, which could be related to land-based contaminants' sources. The influence of natural factors and other contaminants (e.g., pharmaceuticals) on biomarkers' responses are also discussed., (Copyright © 2010 Wiley Periodicals, Inc.)

Richards Bay Harbour is South Africa's premier bulk cargo port. It was constructed in the Mhlathuze estuary in 1976 and over the past 34 years has become South Africa's most modern and largest cargo handling port. Although no official monitoring programme is in progress various studies by different groups have provided relevant data with respect to changing metal levels in brown mussel tissue (Perna perna) over the last 34 years. Eleven elements were analysed in brown mussels from the main channel in Richards Bay Harbour using ICP-MS. The results indicate that the metal concentrations in the mussel tissue remained relatively constant between 1974 and 2005. The mean metal concentrations increased significantly in 2005 possibly due to the construction of the new coal terminal and associated dredging activities. Mean metal concentrations in the 2008 sampling event were also elevated due to increased run off during an above average rainy season., (Copyright © 2011. Published by Elsevier Ltd.)

Oxidative stress indices were measured in gills and digestive glands of Perna viridis collected from three coastal locations in Goa i.e., Bambolim, Marmugao Harbour and Malim. In addition to lipid peroxidation, the activities of antioxidant enzymes such as superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase and glutathione S-transferase and two non-enzymatic antioxidants (ascorbic acid and reduced glutathione) were investigated in order to understand their variation with respect to pollution status of the sampling locations. We observed a significant increase in lipid peroxidation and antioxidant enzymes of both the tissues at Mormugao Harbour and Malim, suggesting that the animals at these two locations are at higher level of oxidative stress as compared to those at Bambolim. Conversely, low levels of non-enzymatic antioxidants such as ascorbic acid and reduced glutathione, observed at Mormugao Harbour and Malim indicate that the animals may use these compounds to counteract stress in the tissues. This study shows that changes in lipid peroxidation, superoxide dismutase, catalase, glutathione reductase, glutathione S-transferase and reduced glutathione in tissues of P. viridis can be used as molecular biomarkers in environmental monitoring programs.

Perna viridis is an ideal animal for studying the impairment caused by the effects of heavy metals that are often detected in coastal areas. Preliminary bioassay tests revealed that the lethal (LC(100)), median lethal (LC(50)) and sublethal (LC(0)) concentration of silver and chromium to P. viridis were 6.5, 4.0, 2.0 mg l(-1) and 4.5, 2.5, 1.0 mg l(-1), respectively. Toxic effect of silver and chromium was evaluated in the green mussel P. viridis, with reference to oxygen consumption, filtration rate and ATPase system in laboratory experiments. These parameters were selected as the end point of sublethal stress. Oxygen consumption and filtration rates were calculated as a measure of decline in the dissolved oxygen level and algal concentration (feed) in the aquaria water, respectively. Silver and chromium affects both oxygen consumption and filtration rate significantly (P<0.01) at 96 h when compared to control. The activity of ATPases system in the gills, hepatopancreas, ovary and muscle of mussels were inhibited by silver and chromium indicating that metals exerted significant toxic effect. The inhibition of Na(+)K(+) ATPase, Ca(2+) ATPase and Mg(2+) ATPase in the mussels were significant (P<0.05) for silver and highly significant (P<0.01) for chromium, which indicates that chromium was more toxic to mussels when compared to silver. The assessment of oxygen consumption, filtration and ATPases system can thus be used as a valid biomarker in aquatic ecotoxicology studies.

The mussel Perna perna was used to monitor and assess areas suspected of oil contamination by transplanting animals from unimpacted to impacted sites and vice versa. The deployment lasted for three months during which total PAH were measured in seawater samples collected every 15 days. PAH concentrations were determined in mussel tissue monthly. The lysosomal neutral red assay and the condition index were used as biomarkers of the adverse biological effects. The PAH concentration in mussels at unimpacted sites, prior to deployment was ca. 100 microg kg(-1) but increased to 300 microg kg(-1) after three months at the contaminated site. Mussels transplanted to the uncontaminated site showed a decrease from 380 to 80 microg kg(-1). Significant changes in lysosomal stability occurred within one month of the beginning of the transposition; in contrast condition indexes were not correlated with the observed changes in PAH concentration.

Environmental pollution is a growing concern and, more importantly, pollution of the aquatic ecosystem is alarming. Marine pollution may be one of the reasons for disease incidence in marine organisms, which is caused due to adverse effects of pollutants on the immune system. Bivalves are commonly used as bio-indicators of marine pollution, and immunomodulation due to toxicants is one of the important bio-markers used. Perna viridis too have been used as a bio-indicator, but this study is, to our knowledge, a first report on immunomodulation produced by metals, in P. viridis. Animals were exposed to copper and mercury at their sub-lethal concentrations of 20 microg L(-1) and 10 microg L(-1), respectively. Immune parameters including phenoloxidase, reactive oxygen species generation, and phagocytosis were monitored. The study period was for 25 days (chronic long-term exposure) and objectives established whether metals produced immunomodulation and to understand the effects of long-term exposure on immunomodulation. Results showed that both metals adversely affected immune parameters studied and, interestingly, there appears to be some level of recovery (depuration) from the toxic effects of metals.

Various antifouling biocides were surveyed in sediment and green mussels (Perna viridis) from the coastal area of Thailand. The concentrations of butyltin (MBT), dibutyltin (DBT), and tiributyltin (TBT) in sediment from Thailand were in the range of 1-293 microg kg(-1) dry wt., 1-368 microg kg(-1) dry wt., and 2-1246 microg kg(-1) dry wt., respectively. A higher concentration of TBT was observed in industrial areas, where many tankers and cargo ships sail and moor. Phenyltin (PT) compounds were not detected in most of the sampling sites. The concentrations of MBT, DBT, and TBT in green mussels from Thailand were in the range of 8-20 microg kg(-1) wet wt., 4-9 microg kg(-1) wet wt., and 4-45 microg kg(-1) wet wt., respectively. The concentration of TBT was high in an area where aquaculture practice is common. The detection frequencies of TPT were low. Representative booster biocides were surveyed in sediment. The detection frequencies of Sea Nine 211 were low (2/13). Furthermore, Sea Nine 211 concentrations in the detected samples were at values near the detection limit (0.051-0.094 microg kg(-1) dry wt.). Diuron was detected at the highest level among the booster biocides in the range of 0.07-25 microg kg(-1) dry wt. Irgarol 1051 was detected in the range of 0.03-3.2 microg kg(-1) dry wt., and concentrations of the degradation product's M1 were 0.03-4.9 microg kg(-1) dry wt. Diuron and Irgarol 1051 showed higher concentrations in industrial areas and fishery sites, respectively. Diuron was only detected among green mussels in the range of <0.64-9.6 microg kg(-1) wet wt. The lower ratio of Diuron and Irgarol 1051 for TBT concentration suggests that TBT is still being used in Thailand.

The green mussel Perna viridis was sampled from relatively clean and contaminated sites along the Kartanata-Kerala coast (south west coast of India) to study the tissue concentration of trace metals and biological responses to stress (biomarkers) such as sister chromatid exchange (SCE), chromosomal aberration, micronucleus (MN) test, hemic neoplasia (HN), Chromotest (Ames test) and comet assay. In general, mean tissue concentrations of toxic trace metals collected from 25 sampling sites were found to be below the World Health Organisation (WHO) permissible concentration given for seafood. The digestive gland extract of mussels from all 25 sampling sites showed negative reaction for mutagenic activity (Ames test) in the absence of metabolic activation. Very low levels of chromosomal aberration, SCE, MN, HN and comet cells were observed in mussels collected from both the urban associated and relatively clean sites. This study seems to indicate that that the coastal waters of Karnataka and Kerala are minimally contaminated with genotoxic and carcinogenic chemicals.