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Chondroitin sulfate (CS) and heparan sulfate (HS) are sulfated glycosaminoglycan (GAG) chains that consist of repeating disaccharide units composed of hexosamine and hexuronic acid. GAG chains exhibit diverse bioactivities in a structure-specific manner. Marine invertebrates are a rich source of highly sulfated and rare structures of GAG chains. Here, we isolated GAGs from the green-lipped mussel Perna canaliculus, an aquaculture species that is produced on a large scale. We separated GAGs based on the degree of negative charges and analyzed their disaccharide compositions. CS and HS both exhibited characteristic compositions of differently sulfated disaccharides. CS chains showed a higher degree of sulfation than HS chains and contained a high percentage of the E unit disaccharide GlcA-GalNAc(4,6-O-disulfate). Furthermore, CS chains rich in the E unit stimulated the neurite outgrowth of primary cultured neurons. The present results indicate the potential of P. canaliculus GAGs as biomaterials to study the structure-function relationships of GAGs.

Despite the nutritional benefits, bivalves like mussels are also an excellent aquatic heavy metal biomonitoring agent due to their high tolerance to varying levels of temperature, salinity and oxygen, as well as pollutants. Although the accumulated toxic heavy metals may not exert direct negative impacts on the mussels, such toxicants in mussel tissues can give harmful effects on human body when consumed in toxic quantities and/or over prolonged period. The booming of urban and industrial activities, and consequently the increment of runoffs, as well as wastewater effluents and leaching, further exacerbated the magnitude of this issue. Hence, continuous monitoring of heavy metal contents in mussels is vital to ensure its compliance with food safety regulations, protecting consumers at large. This review paper discusses the occurrence of heavy metals in mussels especially that of Perna viridis in Malaysia and other parts of the world since year 2000 until 2021. Heavy metal concentration data and patterns from various coastal and/or estuaries were compared. Where applicable, statistical data that indicate variations between sampling sites, sampling months or years and chemical treatments for heavy metal removal were critically reviewed. Health risk assessment findings were also discussed. More importantly, related chemical-based interventions to minimize and/or eliminate toxic heavy metals from mussels are also reviewed.

Understanding of pathogenicity and immunity is crucial in producing disease-resistant cultured mollusk varieties. This study aimed to isolate pathogenic Vibrio alginolyticus from naturally infected Perna viridis, and to determine histopathological and immunological changes after challenge test with the same bacteria. Biochemical tests and 16S rDNA identified the pathogen as V. alginolyticus (99%). Antibiotic susceptibility test showed ampicillin resistance of the pathogen. Pathogenicity assay was conducted by immersing P. viridis in 1.5 × 106 CFU mL−1 V. alginolyticus for 60 min and observed for 5 days. Clinical signs, histopathological and immunological alterations were observed and monitored. Infected groups showed 60% mortality and decreased immunity factors, including total hemocyte count and lysozymes activity. Histopathological examination revealed pathological lesions in the hepatopancreas at 24 h post-challenge and hemocyte proliferation as part of a severe inflammatory reaction. Karyomegaly in the hepatopancreas tissue, concomitant with necrosis demolition of tubules cells, was also observed. V. alginolyticus was determined to be pathogenic to P. viridis, causing mortality as a result of multiple organ lesions and dysfunction in digestive gland and immune organs. This study demonstrated the role of histopathological and immunological parameters as potential biomarkers in assessing vibriosis caused by Vibrio species in green mussel, P. viridis.

Elevated metal concentrations can become harmful to marine organisms and to humans that consume them. Metal concentrations at multiple sites around Algoa Bay, South Africa, were last investigated in the 1980s. We collected wild brown mussels (Perna perna) from seven sites around Algoa Bay, and quantified metallic elements using ICPMS. Metallic element concentrations differed significantly among the sampling sites and correlated with pollution sources at specific sites. The concentration of Pb in mussels at one site slightly exceeded South African limits. Based on the South African estimated daily intake, the target hazard quotient, and South African metal limits, mussels from Algoa Bay are safe for human consumption, except possibly from one site. However, combined with data on bisphenols and benzophenone UV filters in P. perna from the same sites, we suggest a possible health concern to consumers.

In the framework of the biomonitoring programme of the Gulf of Annaba (north-eastern Algeria), this study aims to assess the health status of the Gulf by transplanting the brown mussel Perna perna (Linnaeus 1758) for 12 weeks (June 2017-August 2017) at three sites. The concentrations of copper (Cu), zinc (Zn), lead (Pb) and cadmium (Cd) and as well as a battery of enzymatic and non-enzymatic biomarkers were measured in mussels before and after the transplantation period. Furthermore, analysis of trace metals was performed on the surface layer of the sediment of all the sites. A significant increase in the Cu and Pb concentrations in the mussel dry tissues was observed after the transplantation period and followed the order of metal contamination of the surface layer sediments, indicating a relationship between the bioaccumulation of metals and their bioavailability at each site. Unlike those of Cu and Pb, Cd and Zn concentrations did not reach significant levels of bioaccumulation at any of the three study sites. The biomarker response results were complementary to the measured metal concentrations in the mussel tissues and were associated with the metal accumulation index. Metal bioaccumulation in mussels and supporting biomarker response results identified the most important pollution point sources in the area.

Considering the importance of hemocyte characterization for immunological studies, this work aimed to characterize the hemocyte types of Perna perna mussels combining transmission electron microscopy and flow cytometry with the classical optical microscopy. The results indicated four type of hemocytes: hyalinocytes, semigranulocytes, granulocytes and blast-like cells.

Solution structure of byssal plaque protein Pvfp-5 beta secreted by the Asian green mussel Perna viridis gives molecular insight into mussel adhesion on wet surfaces.Some marine organisms can resist to aqueous tidal environments and adhere tightly on wet surface. This behavior has raised increasing attention for potential applications in medicine, biomaterials, and tissue engineering. In mussels, adhesive forces to the rock are the resultant of proteinic fibrous formations called byssus. We present the solution structure of Pvfp-5 beta, one of the three byssal plaque proteins secreted by the Asian green mussel Perna viridis, and the component responsible for initiating interactions with the substrate. We demonstrate that Pvfp-5 beta has a stably folded structure in agreement with the presence in the sequence of two EGF motifs. The structure is highly rigid except for a few residues affected by slow local motions in the mu s-ms time scale, and differs from the model calculated by artificial intelligence methods for the relative orientation of the EGF modules, which is something where computational methods still underperform. We also show that Pvfp-5 beta is able to coacervate even with no DOPA modification, giving thus insights both for understanding the adhesion mechanism of adhesive mussel proteins, and developing of biomaterials.

Mussels, which occupy important positions in marine ecosystems, attach tightly to underwater substrates using a proteinaceous holdfast known as the byssus, which is tough, durable, and resistant to enzymatic degradation. Although various byssal proteins have been identified, the mechanisms by which it achieves such durability are unknown. Here we report comprehensive identification of genes involved in byssus formation through whole-genome and foot-specific transcriptomic analyses of the green mussel, Perna viridis. Interestingly, proteins encoded by highly expressed genes include proteinase inhibitors and defense proteins, including lysozyme and lectins, in addition to structural proteins and protein modification enzymes that probably catalyze polymerization and insolubilization. This assemblage of structural and protective molecules constitutes a multi-pronged strategy to render the byssus highly resistant to environmental insults.

Perna perna mussels, abundant throughout the Brazilian coast, are routinely applied as bioindicators in environmental monitoring actions due to their sessile and filter-feeding characteristics. In addition, they are noteworthy for their food importance, especially for coastal populations. In this context, the aim of this study was to investigate elemental contamination in commercially marketed and highly consumed P. perna samples from the highly impacted Guanabara Bay, Rio de Janeiro, Brazil. A total of 30 mussels were sampled, and elemental concentrations (As, Cd, Cr, Cu, Fe, Hg, Mn, Ni, Pb, Se, V, and Zn) were determined in adductor muscle samples by inductively coupled plasma mass spectrometry (ICP-MS). Human consumption risks were assessed by comparisons to Brazilian and international legislations. No significant differences between sex were observed for all analyzed elements. Even when analyzing only the adductor muscle, all mussel samples exceeded the Brazilian limit for Cr, while 12 samples exceeded the limit for Se. When compared to other regulatory agencies, As and Zn levels were higher than the limits set by China, New Zealand, and the USA. Estimated daily dietary intake values were not above limits imposed by the Food and Agriculture Organization of the United Nations/World Health Organization for any of the assessed elements, although it is important to note that only the adductor muscle was assessed. Therefore, continuous metal and metalloid monitoring in bivalves in the study region is suggested, as metal transport and bioavailability, especially in coastal estuaries such as Guanabara Bay, which are currently undergoing significant changes due to anthropogenic activities.

The New Zealand Greenshell™ mussel (Perna canaliculus) is an endemic bivalve species with cultural importance, that is harvested recreationally and commercially. However, production is currently hampered by increasing incidences of summer mortality in farmed and wild populations. While the causative factors for these mortality events are still unknown, it is believed that increasing seawater temperatures and pathogen loads are potentially at play. To improve our understanding of these processes, challenge experiments were conducted to investigate the combined effects of increased seawater temperature and Vibrio infection on the immune and metabolic responses of adult mussels. Biomarkers that measure the physiological response of mussels to multiple-stressors can be utilised to study resilience in a changing environment, and support efforts to strengthen biosecurity management. Mussels acclimated to two temperatures (16 °C and 24 °C) were injected with either autoclaved, filtered seawater (control) or Vibriosp. DO1 (infected). Then, haemolymph was sampled 24 h post-injection and analysed to quantify haemocyte immune responses (via flow-cytometry), antioxidant capacity (measured electrochemically) and metabolic responses (via gas chromatography-mass spectrometry) to bacterial infection. Both seawater temperature and injection type significantly influenced the immune and metabolite status of mussels. A lack of interaction effects between temperature and injection type indicated that the effects of Vibrio sp. 24 h post-infection were similar between seawater temperatures. Infected mussels had a higher proportion of dead haemocytes and lower overall haemocyte counts than uninfected controls. The proportion of haemocytes showing evidence of apoptosis was higher in mussels held at 24 °C compared with those held at 16 °C. The proportion of haemocytes producing reactive oxygen species did not differ between temperatures or injection treatments. Mussels held at 24 °C exhibited elevated levels of metabolites linked to the glycolysis pathway to support energy production. The saccharopin-lysine pathway metabolites were also increased in these mussels, indicating the role of lysine metabolism. A decrease in metabolic activity (decreases in BCAAs, GABA, urea cycle metabolites, oxidative stress metabolites) was largely seen in mussels injected with Vibrio sp. Itaconate increased as seen in previous studies, suggesting that antimicrobial activity may have been activated in infected mussels. This study highlights the complex nature of immune and metabolic responses in mussels exposed to multiple stressors and gives an insight into Vibrio sp. infection mechanisms at different seawater temperatures.

Microplastics are ubiquitous in the marine environment, and their uptake by many organisms has been well documented. Concern about increasing plastic waste in ecosystems and organisms has led to the production of biodegradable alternatives. However, long breakdown times of biodegradable plastics in natural environments mean they still have the potential to induce ecological impacts. The impacts of microplastics on organisms remain unclear, especially as many experimental microplastic exposures employ particle concentrations orders of magnitude greater than those found in natural ecosystems. Here, we exposed the ecosystem engineer, the Asian green mussel Perna viridis, to non-biodegradable and biodegradable microplastics at two environmentally relevant concentrations (~17-20 particles L

A period of seismic activity starting in 2010 coincided with a decline in commercial catches of wild seed mussels in a major aquaculture production region of New Zealand. Analyses of over 40 years of mussel seed catch data from in the Pelorus and Kenepuru Sounds, confirmed a marked decline since 2010 in catches of the preferred, green-lipped mussel (Perna canaliculus), the larvae of which is known to have low tolerance of heavy metals in seawater.Heavy metal mean concentrations were measured throughout the Pelorus and Kenepuru Sounds. The concentrations ranged from 0.60-3.24, 16.94-74.35, 1.47-4.00, 2.23-19.02, 1.86-3.29 and 0.12-0.52 µg LNo mussel embryos survived when incubated in these seawater samples. The mean concentrations of Cr, Fe, As, and Cd were significantly higher in the seawater from the Sounds than in the hatchery seawater. A higher concentration of one or a combination of these heavy metals could be the cause of the poor larval survival. These findings could be crucial for the sustainability of mussel farming in the area.

Green-lipped mussels (Perna canaliculus) are a commercially and culturally important bivalve species in New Zealand (NZ). As the highest value export aquaculture product in NZ, understanding and safeguarding the health of this species is imperative. The identification and characterization of hemocytes can provide useful information regarding the health of this species. Using flow cytometry (FCM), the present study assessed for the first time the use of different antiaggregant solutions and storage times on the immune-related parameters of hemocytes from cultured adult P. canaliculus. In addition, characterization of the immune-related functions of hemocyte sub-populations within the hemolymph were assessed. The two antiaggregant solutions tested (Modified Alserver's, MAS, A and B) maintained similar numbers of hemocytes in circulation over a 60 min period but, reduced the viability (MAS A) and increased the ROS production (MAS B) of the hemocytes compared to hemocytes diluted in cold filtered seawater (FSW). Hemocytes diluted in FSW and kept on ice showed significant aggregation after 2 h and a reduction in viability from 4 h. Three different hemocyte sub-populations were identified, discernible by their relative size and internal complexity: blast-like cells, hyalinocytes and granulocytes, which accounted for approximately 4, 67 and 29% of the total hemolymph population respectively. Granulocytes showed significantly higher reactive oxygen species production, phagocytic capabilities and neutral lipid content compared to hyalinocytes and blast-like cells. Results indicate that maintaining extracted hemolymph in cold FSW, completing analysis of fresh samples within 2 h of extraction and FCM assay incubation times of no longer than 30 min are best to obtain accurate results. Formalin fixation can also be used for future determination of hemocyte sub-populations and internal structures. Results from this study will allow effective future study of the effects of various stressors on P. canaliculus health and lead to improved management and production strategies in this species.

Due to the ever-increasing production of plastic litter and its subsequent accumulation as microplastic in the environment, the pollution caused by microplastics is considered as a global menace, especially in the coastal ecosystem. Occurrence of microplastics in water and three commercially important bivalves, Viz. green mussel (Perna viridis), edible oyster (Magallana bilineata) and black clam (Villorita cyprinoides) from five different locations of southwest coast of India was studied. The highest abundance of microplastics was observed in water samples from Periyar River (163.67 items L

Poor health and mortality events of the commercially important and endemic New Zealand green-lipped mussel (Perna canaliculus) pose a threat to its industry. Despite the known importance of microbiomes to animal health and environmental resilience, the host-associated microbiome is unexplored in this species. We conducted the first baseline characterization of bacteria and fungi within key host tissues (gills, haemolymph, digestive gland, and stomach) using high-throughput amplicon sequencing of 16S rRNA gene and ITS1 region for bacteria and fungi, respectively. Tissue types displayed distinctive bacterial profiles, consistent among individuals, that were dominated by phyla which reflect (1) a fluid exchange between the circulatory system (gills and haemolymph) and surrounding aqueous environment and (2) a highly diverse digestive system (digestive gland and stomach) microbiota. Gammaproteobacteria and Campylobacterota were mostly identified in the gill tissue and haemolymph, and were also found in high abundance in seawater. Digestive gland and stomach tissues were dominated by common gut bacterial phyla, such as Firmicutes, Cyanobacteria, Proteobacteria, and Bacteroidota, which reflects the selectivity of the digestive system and food-based influences. Other major notable taxa included the family Spirochaetaceae, and genera Endozoicomonas, Psychrilyobacter, Moritella and Poseidonibacter, which were highly variable among tissue types and samples. More than 50% of fungal amplicon sequence variants (ASVs) were unclassified beyond the phylum level, which reflects the lack of studies with marine fungi. However, the majority of those identified were assigned to the phylum Ascomycota. The findings from this work provide the first insight into healthy tissue microbiomes of P. canaliculus and is of central importance to understanding the effect of environmental changes on farmed mussels at the microbial level.

Accumulation of microplastics (MP) in oceanic waters is eroding the health of marine biota. We investigated how size-fractionated MP influence the toxicity risks towards a tropical keystone species, Perna viridis. Tissue-specific bioaccumulation and in vivo toxicity of polystyrene (PS) particles (0.5, 5, and 50 μm) were measured upon continuous exposure for 7 days, followed by 7 days depuration. P. viridis were exposed to equivalent mass (0.6 mg/L), corresponding to 4.0-4.6 particles/mL, 4.6-7.1 × 10

Invasive, fouling species increase management costs and reduce mussel growth, which jeopardizes mariculture. We studied the distribution of eight invasive species in Santa Catarina, the leading mussel producer in Brazil. Our goals were to determine their spatial distribution and prevalence on farm structures (buoys, long lines, and mussel socks), as well as understand the relevance of propagule pressure (recruitment), port distance, and area of the farm in this distribution. Although present in all sites, adult and recruits distribution were spatially restricted, showing that species might have a metapopulation structure. The most prevalent species were the ascidian Styela plicata, the barnacle Megabalanus coccopoma, the bryozoan Schizoporella errata, and the polychaete Branchiomma luctuosum. Recruitment was the main driver of three species distribution while distance to port explained only one species distribution. Based on those results, we discuss policy options, management, and regulation enforcement, that can be used in the mussel aquaculture elsewhere.

Stress and survival of the juvenile New Zealand green-lipped mussel, Perna canaliculus, is a poorly understood bottleneck in the ecological and economic performance of a significant aquaculture crop. This species was therefore selected as a model organism for the development of a new method to quantify oxidative stress in whole individuals. An in vivo ROS-activated stain (CellROX™) was administered to anaesthetised, translucent juveniles that were subsequently formaldehyde fixed and then visualised using confocal microscopy. Subsequent application of image analysis to quantifying ROS-positive tissue areas was successfully used to detect stress differences in juvenile mussels exposed to varying levels of emersion. This integrated method can be used to localise and quantify ROS production in individual translucent bivalve life stages (larval and juvenile), while relative stability following fixation greatly expands potential practical field applications. This article has an associated First Person interview with the first and third authors of the paper., Summary: A new method to quantify reactive oxygen species in live mussels using image analysis. The unusual fixation step allows for in situ staining, followed by image analysis in the laboratory.

Mass mortalities of New Zealand Green-lipped mussels (Perna canaliculus) are thought to be associated with increased water temperatures and immune challenges from opportunistic pathogens. However, the combined effects of acute thermal stress and immune stimulation on mussels are poorly understood. To investigate these responses, adult mussels were exposed to different temperatures (26 °C [thermal stress] vs 15 °C [ambient]) and a bacterial-derived endotoxin injection (with vs without) to mimic a pathogen infection. Various immunological and metabolic parameters were measured over two days via enzyme staining reactions, flow cytometry, and metabolomic profiling. None of the treatments impacted total and differential haemocyte counts, haemocyte viability or production of reactive oxygen species. Acid phosphatase and phenoloxidase activities were detected only within granulocytes (not in hyalinocytes), although their relative expressions also were not affected. Conversely, metabolite profiling exposed impacts of thermal stress and endotoxin exposure at a metabolic level, indicative of physiological changes in energy expenditure and partitioning. At the higher water temperature, free fatty acid and amino acid constituents increased and decreased, respectively, which supports an elevated energy demand and higher metabolic rate due to thermal stress. Ultimately, energy production is being sustained via multiple routes including the glycolysis pathway, TCA cycle, and β-oxidation. Additionally, branched-chain amino acids, the urea cycle and the glutathione pathway were affected by the higher temperature. The metabolic response of mussels exposed to endotoxin exposure resulted in increased metabolite response largely linked to protein and lipid degradation. After 5 days of exposure, survival data confirmed a severe physiological impact of the higher temperature through incidences of mortality. However, the thermal challenge in combination with the specific endotoxin treatment applied did not lead to a synergistic effect on mortality. These findings provide new insights into the relationship between thermal stress and immunity to better understand the immune defence system in mussels.

Mytilin is one of the most important CS-αβ peptides involved in innate immune response in Mytilidae. In this study, we successfully identified four mytilin-like antimicrobial peptides (pernalins) from Asian green mussel Perna viridis by aligning the P. viridis transcriptome with 186 mytilins and myticins related sequences collected from the transcriptome data of six Mytilus species. Analysis on gene structure showed that pernalin genes had high conservation with mytilin B of Mediterranean mussel Mytilus galloprovincialis. Interestingly, all pernalin genes have a similar tissue expression feature, evidenced by the highest transcription level observed in the hemocytes and followed by the mantle. The lowest transcription level was observed in the foot and gills. qRT-PCR analysis showed that all pernalin genes were significantly down-regulated at each time points from 3 h to 48 h after Vibrio parahaemolyticus infection, suggesting their timely immune responses after bacterial infection.

Diarrheic shellfish poisoning (DSP) toxins are widely distributed over the world, causing diarrhea, vomiting, and even tumor in human. However, bivalves, the main carrier of the DSP toxins, have some tolerant mechanisms to DSP toxins, though it remains unclear. In this study, we scrutinized the role of Jun N-terminal kinases (JNK) in tolerance of DSP toxins and the relationship between JNK, apoptosis and nuclear factor E2-related factor/antioxidant response element (Nrf2/ARE) pathways. We found that the phosphorylated level of JNK protein was significantly increased both in hemocytes (6 h) and gills (3 h) of the mussel Perna viridis after short-term exposure to DSP toxins-producing dinoflagellate Prorocentrum lima. Exposure of P. lima induced oxidative stress in mussels. Hemocytes and gills displayed different sensitivities to the cytotoxicity of DSP toxins. Exposure of P. lima activated caspase-3 and induced apoptosis in gills but did not induce caspase-3 and apoptosis in hemocytes. The short-term exposure of P. lima could activate Nrf2/ARE signaling pathway in hemocytes (6 h), while longer-term exposure could induce glutathione reductase (GR) expression in hemocytes (96 h) and glutathione-S-transferases (GST) in gills (96 h). Based on the phylogenetic tree of Nrf2, Nrf2 in P. viridis was closely related to that in other mussels, especially Mytilus coruscus, but far from that in Mus musculus. The most likely phosphorylated site of Nrf2 in the mussels P. viridis is threonine 504 for JNK, which is different from that in M. musculus. Taken all together, the tolerant mechanism of P. viridis to DSP toxins might be involved in JNK and Nrf2/ARE signaling pathways, and JNK play a key role in the mechanism. Our findings provide a new clue to further understand tolerant mechanisms of bivalves to DSP toxins.

To develop a dried rice flour powder (DP) formulation to contain a lactic acid bacterial starter culture for fermenting mussel meat (FM).Lactiplantibacillus plantarum KU-LM173 (LP), Enterococcus hirae KU-LM174 and Pediococcus acidilactici KU-LM145 (PA) were selected from commercial FMs and identified to have high acid and protease production. Mixed culture between LP, for high acid production, and PA, for the flavour, was the best for DP and had greater organoleptic properties than a single starter fermentation. The best ratio of DP for production was 1% of the mussel weight, while the highest numeric scoring of the organoleptic test between 3% and 6%. The starter culture fermentation accelerated over the natural (wild) fermentation and ended at day 3. The shelf life of the product was at least 30 days at 30-35°C with no pathogens detected. The shelf life of DP at 4°C was 10 weeks.DP with the best strains and long shelf life promoted safety of FM and reduced the processing time. High consumer acceptance, protease and acid production and flavour were unique product characteristics.Accelerated commercial FMs with effective DP formulation for the industrial sector may be plausible.

Bivalve molluscs have been regarded as excellent bioindicators of environmental pollution as they persistently accumulate toxic contaminants present in their ecosystem. Histological alterations in the digestive gland and gills of three bivalve sp., Viz. edible oyster (Magallana bilineata), green mussel (Perna viridis) and black clam (Villorita cyprinoides) from ecologically sensitive regions of international significance on the southwest coast of India were evaluated using a semi-quantitative histopathological index to assess the environmental quality. The prominent tissue alterations included tubular vacuolation, haemocytic infiltration, parasitosis, lamellar disorganization, and the presence of prokaryotic inclusions. The presence of ten trace metals was also evaluated in the digestive gland of bivalves. The histopathological indices were evaluated season-wise and region-wise. Seasonal variation in all the reaction patterns was observed in the digestive gland across sampling zones, with the highest indices observed during post-monsoon. The indices for all the reaction patterns in the digestive gland were significantly higher in bivalves from Vembanad Lake (Z4), followed by Periyar River (Z5). The indices for cellular changes and parasitosis in gills were the highest in the Ashtamudi estuary (Z1) and Z5, respectively. The global histopathological indices of the digestive gland and gills were also the highest in Z4, followed by Z5. Principal component analysis revealed that Z4 was distinct with the highest metal pollution index. A positive relation was observed with heavy metals, digestive gland histological alterations, and season and region of sampling.

In the past decade, the use of marine mussels as seafood is being more popular. They considered being a rich source of various nutritional bioactive compounds that aroused the scientific community's interest. This study investigated the antioxidant and the antithrombotic consequences on Sprague-Dawley male rats after adding dried New Zealand mussel Perna canaliculus in their diet. The biochemical, hematological and histopathological changes were also observed. Forty rats were divided into four groups according to the amount of dried mussels in their diet, in addition to a control group that consumed a basal diet only. Group 1 consumed 25% dried mussels in its basal diet; Group 2 consumed 35% dried mussels in its basal diet, and Group 3 was consumed 45% dried mussels in its basal diet. The biochemical parameters showed improvements in liver function. Interestingly, the lipid profile decreased, especially the low-density lipoprotein cholesterol (LDL-C) levels which were reduced significantly in Group 3 (p < .01). These observations were accompanied by a decrease in iron levels significantly as the amount of dried mussels increased (p < .01). Furthermore, the noticed changes in the hematological profile prove that there is an increase in antithrombotic activity. Dried mussels had potent antioxidant effects in the liver as shown by increased lipid peroxide (p < .05), reduced glutathione (p < .05), and glutathione peroxidase (GSH-Px; p < .05). Additionally, antioxidant activity in the kidney was shown to increase through GSH-Px activity (p < .01). In conclusion, these results indicate that consuming dried mussels resulted in improved biochemical and antioxidants activities and could be used as an antithrombotic agent.

Diarrheal shellfish toxins (DSTs) are among the most widely distributed phytotoxins, and are associated with diarrheal shellfish poisoning (DSP) events in human beings all over the world. Therefore, it is urgent and necessary to identify an effective method for toxin removal in bivalves. In this paper, we found that curcumin (CUR), a phytopolylphenol pigment, can inhibit the accumulation of DSTs (okadaic acid-eq) in the digestive gland of Perna viridis after Prorocentrum lima exposure. qPCR results demonstrated that CUR inhibited the induction of DSTs on the aryl hydrocarbon receptor (AhR), hormone receptor 96 (HR96) and CYP3A4 mRNA, indicating that the CUR-induced reduction in DSTs may be correlated with the inhibition of transcriptional induction of AhR, HR96 and CYP3A4. The histological examination showed that P. lima cells caused severe damage to the digestive gland of P. viridis, and the addition of curcumin effectively alleviated the damage induced by P. lima. In conclusion, our findings provide a potential method for the effective removal of toxins from DST-contaminated shellfish.

Perna viridis (Linnaeus, 1758), the Asian green mussel, is native to the Asia-Pacific region. The species is extensively distributed in the Indian subcontinent and is a candidate species for aquaculture in the Southeast Asian region. Availability of genetic information on wild populations is essential for the effective conservation and management of Perna species. The present study assessed the genetic variation and population structure across the distribution range of this species from the Indian peninsula by using microsatellite markers to determine the genetic structuring among the species.A total of 15 microsatellite loci with M13 labeling were used for the genetic characterization of P. viridis along Indian waters. Genotyped data were analyzed using analytical software to determine the genetic stocks and understand the genetic variability across the populations.We identified 15 polymorphic markers to understand the genetic stocks and variability across Perna populations. The mean value of the observed heterozygosity (Hobs: 0.741) for all populations was closer to the expected heterozygosity (Hexp: 0.75). The pairwise Fst values between the west and east coasts of India varied significantly, indicating the existence of significant genetic structure between the populations.Genetic stock identification using software analysis exhibited two distinct stocks, one along the west coast (Arabian Sea) and another along the east coast (Bay of Bengal). Bottleneck analysis indicated the genetic stability of species in the wild. P. viridis is a commercially vital species in Indian peninsular regions. The present study suggests the adoption of stock-specific relaying programs of the species from Indian waters in future studies.

During their lifecycle, many marine organisms rely on natural adhesives to attach to wet surfaces for movement and self-defence in aqueous tidal environments. Adhesive proteins from mussels are biocompatible and elicit only minimal immune responses in humans. Therefore these proteins have received increased attention for their potential applications in medicine, biomaterials and biotechnology. The Asian green mussel Perna viridis secretes several byssal plaque proteins, molecules that help anchor the mussel to surfaces. Among these proteins, protein-5β (Pvfp-5β) initiates interactions with the substrate, displacing interfacial water molecules before binding to the surface. Here, we established the first recombinant expression in Escherichia coli of Pvfp-5β. We characterized recombinant Pvfp-5β, finding that despite displaying a CD spectrum consistent with features of a random coil, the protein is correctly folded as indicated by MS and NMR analyses. Pvfp-5β folds as a β-sheet–rich protein as expected for an epidermal growth factor (EGF)-like module. We examined the effects of Pvfp-5β on cell viability and adhesion capacity in NIH-3T3 and HeLa cell lines, revealing that Pvfp-5β has no cytotoxic effects at the protein concentrations used and provides good cell-adhesion strength on both glass and plastic plates. Our findings suggest that the adhesive properties of recombinant Pvfp-5β make it an efficient surface coating material, potentially suitable for biomedical applications including regeneration of damaged tissues.

We report the occurrence and genomic features of multidrug-resistant vancomycin-resistant Enterococcus faecium vanA belonging to a novel sequence type (designated ST1336), carrying a Tn1546-like element, in marine brown mussels (Perna perna) from anthropogenically affected coastal waters of the Atlantic coast of Brazil, highlighting a potential source of dissemination for related ecosystems, with additional consequences for seafood safety and quality.

Plastic pollution is a pervasive problem to marine life. This study aimed (1) to investigate levels of microplastic in wild and farmed mussels (Perna perna), and (2) to assess the effectiveness of depuration in reducing micro plastics. Wild and farmed mussels were sampled from Guanabara Bay (Southwestern Atlantic). Four treatments were compared (N = 10 mussels/treatment): wild non-depurated mussels, wild depurated mussels, farmed non depurated mussels, and farmed depurated mussels. Up to 31.2 +/- 17.8 microplastics/mussel (>= 0.45 pm) were detected (means +/- SD), and microplastics were present in all 40 individuals analyzed. Nylon fibers were more abundant than polymethyl methacrylate (PMMA) fragments. Blue, transparent, and red nylon fibers were more abundant in both wild and farmed mussels. Although 93 h-depuration significantly reduced microplastics (ANOVA, p = 0.02) in both wild (46.79%) and farmed mussels (28.95%), differences between farmed and wild mussels were not significant (p > 0.05). Depuration was more effective in removing blue fibers. Our results highlight the importance of depuration in reducing microplastic pollution in seafood. Brazilian National Research Council (CNPq)National Council for Scientific and Technological Development (CNPq)

There is an increasing concern related to the toxic effects of the soluble portion of diesel oil on aquatic ecosystems and the organisms living in them. In this context, the aim of this study was to analyze the effects of diesel water accommodated-fraction (WAF) on behavioral and biochemical responses of mussels Perna perna. Animals were exposed to 5 and 20% of WAF for 96 h. Prior to the beginning of the experiments, Hall effect sensors and magnets were attached to the valves of the mussels. Valve gaping behavior was continuously recorded for 12 h of exposure and tissues (gills and digestive gland) were separated after 96 h of exposure. Overall, both behavior and biochemical biomarkers were altered due to WAF exposure. Animals exposed to WAF reduced the average amplitude of the valves and the fraction of time opened, and presented greater transition frequency, demonstrating avoidance behavior over the 12 h period. Furthermore, the biochemical biomarkers (GSH, GST, SOD and CAT) were altered following the 96 h of exposure to WAF. Considering the results presented, this study demonstrates the toxic potential of WAF in both shorter and longer exposure periods.

Triazophos (TP) is a widespread pollutant in aquatic environments. A sex-specific metabolic response in green-lipped mussel Perna viridis to TP exposure was observed in our previous study, and this led us to investigate the mechanisms associated with its toxicity. P. viridis were subjected to chronic exposure (15 days) to TP at 35 μg/L to compare the sex-biased transcriptomic profiles in the gonads of male and female mussels. We identified 632 differentially expressed genes (DEGs) (348 up-regulated and 284 down-regulated) in TP-exposed males, and only 61 DEGs (9 up-regulated and 52 down-regulated) in TP-exposed females. Many DEGs were found to be involved in the nervous, reproductive endocrine, oxidative stress, and immune systems of P. viridis. Additionally, enzymatic activity analysis indicated TP induced neurotoxic effects and oxidative damage to the mussels. Our results demonstrate that the stress response and molecular mechanisms of TP toxicology are different between female and male mussels.

Diarrheic shellfish poisoning toxins (DSP toxins) are a set of the most important phycotoxins produced by some dinoflagellates. Studies have shown that DSP toxins have various toxicities such as genotoxicity, cytotoxicity, and immunotoxicity to bivalve mollusks. However, these toxicities appear decreasing with exposure time and concentration of DSP toxins. The underlying mechanism involved remains unclear. In this study, small RNA sequencing was performed in the digestive gland of the mussel Perna viridis after exposure to DSP toxins-producing dinoflagellate Prorocentrum lima for different time periods. The potential roles of miRNAs in response and detoxification to DSP toxins in the mussel were analyzed. Small RNA sequencing of 12 samples from 72 individuals was conducted by BGISEQ-500. A total of 123 mature miRNAs were identified, including 90 conserved miRNAs and 33 potential novel miRNAs. After exposure to P. lima, multiple important miRNAs displayed some alterations. Further miRNA target prediction revealed some important genes involved in cytoskeleton, apoptosis, complement system and immune stress. qPCR demonstrated that miR-71_5, miR-750_1 and novel_mir4 were significantly up-regulated at 6 h after exposure to P. lima, while miR-100_2 was significantly down-regulated after 96 h of exposure. Accordingly, putative target genes of these differentially expressed miRNAs experienced some changes. After 6 h of DSP toxins exposure, NHLRC2 and C1q-like were significantly down-regulated. After 96 h of DSP toxins exposure, NHLRC2 was significantly up-regulated. It is reasonable to speculate that the mussel P. viridis might respond to DSP toxins through miR-750_1, novel_mir4 and miR-71_5 regulating the expression of relevant target genes involved in apoptosis, cytoskeleton, and immune response, etc. This study might provide new clues to uncover the toxic response of bivalve to DSP toxins and lay a foundation for revealing the roles of miRNAs in the environmental adaptation in shellfish.

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Microplastics are prevalent in marine environments and seafood and thus can easily end up in human diets. This has raised serious concerns worldwide, particularly in Hong Kong where the seafood consumption per capita can be three times higher than the global average. This study focused on the green-lipped mussel Perna viridis, a popular seafood species which is subject to a high risk of contamination by microplastics due to its filter-feeding nature. P. viridis was collected from five mariculture sites in Hong Kong and assessed for its body load of microplastics using an automated Raman mapping approach. Microplastics were found in all sites, with an average of 1.60–14.7 particles per mussel per site, or 0.21–1.83 particles per g wet weight. Polypropylene, polyethylene, polystyrene and polyethylene terephthalate were detected among the microplastics, mainly as fragments or fibres in the size range of 40–1000 µm. It was estimated that through consumption of P. viridis, the population in Hong Kong could ingest up to 10,380 pieces of microplastics per person per year. These estimated rates were high compared to the values reported worldwide, suggesting the potential human health risk of microplastics in Hong Kong and adjacent areas.

This study investigates the antioxidant activities of lipid, protein, and carbohydrate extracts from the marine mollusk Perna canaliculus. Lipids were extracted using acetone, which was followed by protein extraction using the broad-spectrum enzyme Alcalase and then carbohydrate extraction using cetylpyridinium chloride. Eighty white BALB/c mice were divided into eight groups according to the administered extracts. Groups 1 and 5 were the control and toxin control groups, respectively. Groups 2, 3, and 4 were administered lipid, protein, and carbohydrate extracts, respectively. The other groups were administered P. canaliculus extracts as well as gentamicin and acetaminophen, known as ethanolic extracts, derived from Nerium oleander to induce oxidation stress. All groups showed significant improvements in body weight (p < 0.05). The lipid extract group showed a significant decrease in low-density lipoprotein cholesterol (p < 0.05) and a significant increase in high-density lipoprotein cholesterol (p < 0.05). After the toxin injection, all groups treated with P. canaliculus extracts showed increased antioxidant effects on hepatocytes (p < 0.05). The lipid extracts induced antioxidant effects to protect the kidney by increasing lipid peroxidation (p < 0.05) and catalase activities (p < 0.05). Also, protein extracts showed antioxidant effects by increasing glutathione and catalase levels significantly (p < 0.005). In conclusion, P. canaliculus extracts, especially lipids and proteins, have potent antioxidant activities that protect vital organs from oxidation stress.

Objectives Intervention studies using New Zealand green-lipped or greenshell™ mussel (GSM) (Perna canaliculus) extract in osteoarthritis (OA) patients have shown effective pain relief. This systematic review summarises the efficacy of GSM extracts in the treatment of OA. Methods A literature search of the three databases EMBASE, MEDLINE, and Scopus was performed to identify relevant articles published up to March 2020. Inclusion criteria were clinical trials published in English measuring the effect of supplementation of whole or a lipid extract from GSM on pain and mobility outcomes in OA patients. Results A total of nine clinical trials were included in systematic review, from which five studies were considered appropriate for inclusion in a forest plot. Pooled results showed that GSM extracts (lipid extract or whole powder) provide moderate and clinically significant treatment effects on a visual analogue scale (VAS) pain score (effect size: − 0.46; 95% CI − 0.82 to − 0.10; p = 0.01). The whole GSM extract improved gastrointestinal symptoms in OA patients taking anti-inflammatory medications. The GSM extract was considered to be generally well tolerated in most of the studies. Conclusion The overall analysis showed that GSM provided moderate and clinically meaningful treatment effects on OA pain. However, the current evidence is limited by the number and quality of studies, and further larger and high-quality studies are needed to confirm the effectiveness and to identify the optimal GSM format. Nevertheless, it is worth considering using GSM extracts especially for patients seeking alternative pain relief treatments with fewer side effects compared to conventional treatment.

Trace metal concentrations were monitored in the yellow clam (Paphia malabarica), green mussel (Perna viridis) and edible oyster (Crassostrea madrasensis) from growing areas in the Ashtamudi and Vembanad estuaries, Kerala. Samples of shellfish (clams n=26, mussels n=18, oysters n=36) and environmental parameters (salinity, temperature, pH and rainfall) were measured in these growing areas from July 2012 to December 2014. Ranges of mean annual concentrations (mg/kg) were Ni (0.46-0.65); Co (2.87-3.49); Fe (80.0-119.4); Mn (3.88-9.38); Zn (40.8-76.2); Pb (1.28-2.00); and Cu (1.59-4.38). In Ashtamudi, clams had higher mean concentrations of Ni, Co, Fe, Mn and Pb than oysters. Mean concentrations of Ni, Pb (in all species), Zn (in clams and mussels) and Cu (in mussels) did not exceed maximum permissible limits mandated by the Food Safety and Standards Authority of India. Mean Mn concentrations exceeded the World Health Organization guideline (1 mg/kg) in the three species while mean Fe concentrations in clams and oysters did not exceed the guideline (100 mg/kg). Target hazard quotients were generally ≤ 1, except for a few Pb results in clams and mussels. Although results suggest no health risk to consumers for the reference doses, daily intakes and elements considered, regular monitoring of trace metals is recommended to maintain consumer protection given increasing anthropogenic and climatic pressures on the shellfish growing areas.

Iron sequestration through ferritin forms a major part of innate immune response in molluscs and detailed understanding of ferritin gene and its functions can be directly applied in infection and disease management studies. Accordingly, identification and detailed molecular characterization of a ferritin subunit gene from a commercially significant marine mussel Perna viridis was targeted. Molecular screening using degenerate primers in total mantle RNA resulted in the amplification of a novel ferritin gene fragment having87% identity to the reported ferritin gene sequences. Rapid amplification of cDNA ends-PCR was followed to generate complete cDNA sequence of P.viridis ferritin (PvFer). The complete cDNA was found to be 798 bp, containing an open reading frame of 522 bp, 5' untranslated region (UTR) of 112 bp and 3' UTR of 165 bp. The 5' UTR and 3' UTR were shown to contain an iron response element (IRE) and a polyadenylation signal (767AATAAA772) with poly (A) tail, respectively. Prediction of stem loop structure revealed that, PvFer-IRE can be folded into a typical secondary stem loop structure, having 5-CAGUGA-3' loop, proximal stem of five paired bases followed by a bulged cysteine, and six nucleotide bottom stem, indicating that expression of PvFer is regulated by iron at the translational level. ORF was found to encode 175 amino acid protein with calculated molecular mass of 19.97 kDa and isoelectric point of 4.97. Examination for signal peptide and phylogenetic analysis confirmed that PvFer belonged to cytosolic ferritins of molluscs. Conserved domain analysis showed that PvFer contained both ferroxidase diiron center and ferrihydrite nucleation center, analogous to ferritin M subunit of bony fishes and amphibians. However, amino acid sequence and glycosylation site showed more homology to vertebrate ferritin H subunits. Predicted 3D models of PvFer resembled the typical spatial features of ferritin proteins. The study forms the first comprehensive identification of a ferritin subunit gene in a true/common mussel (Order: Mytilida). Further, the detailed molecular phylogeny conducted through the present study revealed certain thought provoking insights on ferritin genes of the phylum Mollusca.

The present study has investigated the distribution of microplastics in sediment and its impact on histological, ultrastructural, and oxidative stress mechanisms in Perna viridis (P. viridis) from Kasimedu, Chennai, India. The results confirmed that fibers were the predominant type of microplastics observed, followed by spheres, flakes, sheets, and fragments. The observed microplastics were confirmed as polyester, polypropylene, polyethylene, cellophane, and rayon using μ-FT-IR. Microplastic particles entangled in gills caused abrasion of ciliated structure and hemocyte infiltration in the hemolymph vessels. The digestive gland showed a shrunken nucleus, dark inclusions, and damage in the nucleoid core structure. Enlarged vacuoles and the presence of clusters of vesicles presumably represented the transformed golgi cisternae. Further, the results confirmed that oxidative stress markers were significantly high in gills and digestive diverticula of P. viridis. Overall, the results indicated that microplastics induced different toxic physiological and structural alterations in gills and digestive diverticula of P. viridis. These findings highlighted the necessity to focus on exposure studies to understand the absolute magnitude of the problem due to microplastic pollution in the urban estuarine ecosystems of Chennai, Tamil Nadu, India.

The New Zealand Green-lipped mussel industry is well-established providing vastly to aquaculture exports. To assess mussel health and reproduction status, visual examination of organs and/or collection of haemolymph is commonly applied. Anesthetics, such as magnesium chloride (MgCl2) can be utilized to prevent muscle contraction and keep shells open during sampling. The specific effects of muscle relaxing agents on baseline metabolism in invertebrates is unknown, but it is evident that molecular, cellular and physiological parameters are altered with these chemical applications. To this end, metabolomics approaches can help elucidate the effects of relaxing agents for better assessment of their use as a research tool. Adult Green-lipped mussels were anaesthetized for 3 h in a MgCl2 bath, whereafter haemolymph samples were collected and analyzed via gas chromatography–mass spectrometry applying methyl chloroformate alkylation derivatization. Anesthetized mussels were characterized as non-responsive to manual manipulation, with open valves, and limited siphoning function. Metabolite profiling revealed significant increases in the abundances of most metabolites with an array of metabolic activities affected, resulting in an energy imbalance driven by anaerobic metabolism with altered amino acids acting as neurotransmitters and osmolytes. This research is the first to use a metabolomics approach to identify the metabolic consequences of this commonly used bivalve relaxing technique. Ultimately the use of MgCl2 anesthetization as a sampling strategy should be carefully evaluated and managed when performing metabolomics-related research.

Diarrhetic shellfish toxins (DSTs), some of the most important phycotoxins, are distributed almost all over the world, posing a great threat to human health through the food chain. Therefore, it is of great significance to find effective methods to reduce toxin accumulation in shellfish. In this paper, we observed the effects of four phytochemicals including cinnamaldehyde (CA), quercetin, oridonin and allicin on the accumulation of DSTs in the digestive gland of Perna viridis after exposure to the DSTs-producing Prorocentrum lima. We found that, among the four phytochemicals, CA could effectively decrease the accumulation of DSTs (okadaic acid-eq) in the digestive gland of P. viridis. Further evidence demonstrated that CA could reduce the histological alterations of the digestive gland of a mussel caused by DSTs. RT-qPCR showed that CA could suppress the CYP3A4 induction by DSTs, suggesting that the DSTs&rsquo, decrease induced by CA might be related to the inhibition of CYP3A4 transcription induction. However, further studies on the underlying mechanism, optimal treatment time, ecological safety and cost should be addressed before cinnamaldehyde is used to decrease the accumulation of DSTs in field.

BACKGROUND: Osteoarthritis is a common disease in dogs and cats, and the search for novel treatment options is needed. The combination of green-lipped mussel, curcumin and blackcurrant leaf extract has to date not been studied in dogs and cats. OBJECTIVES: The aim of this study was to test the effect of a supplement containing green-lipped mussel (Perna canaliculus), curcumin (Curcuma longa) and blackcurrant (Ribes nigrum) leaf extract on locomotion and behaviour in client-owned dogs and cats suffering from mild to moderate osteoarthritis. METHODS: To this end, 32 dogs and 16 cats were enrolled in a double-blinded, randomised, crossover, placebo-controlled trial for 10 weeks in cats and 16 weeks in dogs. Outcome parameters were the Helsinki Chronic Pain Index (HCPI) by pet owners in dogs and cats, Canine OsteoArthritis Staging Tool (COAST) by a veterinarian in dogs and Force Plate Analysis (FPA) in 18 dogs. RESULTS: In dogs, the COAST improved significantly in the supplement group compared to baseline but was not different than the placebo group. In cats, the ability to groom, activity level, playfulness and walking up the stairs improved in the supplement group. No differences were found on HCPI scores and FPA in dogs. Several non-responders were noted in both species, which were irrespective of the stage of osteoarthritis. CONCLUSIONS: Overall, the supplement had only partial positive effects in client-owned dogs and cats with mild to moderate osteoarthritis. Further research with a larger sample size and longer duration is needed to expand these findings.

The Greenshell™ mussel (GSM), Perna canaliculus, is a culturally and commercially important species in New Zealand. Declines in spat settlement of GSM have been observed in important growing areas and the cause(s) have not been identified. One hypothesis is that chemical contaminants could be a contributing factor. The aim to this study was to investigate the effects of acute exposure on early life stages using the anti-microbial triclosan (TCS) as a benchmark toxicant and the blue mussel (BM), Mytilus galloprovincialis, as a reference species. Sperm and embryos of BM and GSM were exposed to TCS for 1 h and 48 h, respectively. Following exposures, a range of parameters were investigated including spermatozoa cellular characteristics via flow cytometry, fertilization success, larval mortality and size. Exposure to TCS negatively impacted functional parameters of sperm, reduced the fertilization success and larval size, and increased larval mortality in both BM and GSM with LC50

Bisphenols and benzophenone UV filters are contaminants present in a wide variety of plastic materials and consumer products. The scientific attention towards these contaminants has increased in recent years due to their presence in microplastics, their ubiquitous occurrence in the environment, and their known endocrine disrupting health effects. In this study, the occurrence of nine bisphenol and five benzophenone UV-filter analogues was assessed in wild brown mussels (Perna perna) collected from different sampling sites along the coast of Algoa Bay, South Africa. Eleven out of fourteen target analytes were detected, and bisphenol AP (BPAP) was detected for the first time in mussels, presenting the highest median concentration of 150 ng g