Potential use of gammarus (Pontogammarus maeoticus) and shrimp (Palaemon elegans) as biomonitors of microplastics pollution in coastal environments.

Microplastics (MPs) pose a significant threat to marine ecosystems, necessitating robust biomonitoring to assess aquatic risks and inform effective policymaking. In this study we investigated MPs pollution in gammarus (Pontogammarus maeoticus), shrimp (Palaemon elegans), sediment and water samples of southern coast of the Caspian Sea to assess the potential use of these two crustaceans as biomonitors of MPs pollution, bioconcentration of MPs in organisms' tissue and the pollution risks of MPs in environmental matrices. Samples were collected from 6 stations during June to August 2023. MPs were found in all compartments with an average of 100 ± 45.34 items/kg dry weight, 0.45 ± 0.06 items/L, 0.38 ± 0.21 items/individual or 0.58 ± 0.34 items/g wet weight (ww) and 0.26 ± 0.15 items/individual or 8.69 ± 7.88 items/g ww, for sediments, seawaters, P. elegans and P. maeoticus, respectively. MPs were prevailed by class 300-1000 µm in size, polyamide in polymer, fiber in shape and black in color. P. maeoticus and P. elegans did not meet the selection criteria as MPs biomonitors. However, bioconcentration factor (BCF) illustrated that both crustaceans can absorb and accumulate MPs from their surrounding water (BCF >1). Based on contamination factors (CF) values, sampling stations were polluted with MPs (1 ≤ CF < 6). The overall pollution load index (PLI) for sediment and seawater stations were 2.47 and 1.88, respectively, indicating minor contamination with MPs in the risk level I. Current research provides useful information on MPs pollution in crustaceans species and the risk level of MPs in environmental matrices that can be suitable for bioaccumulation hazard assessment and future monitoring programs.

Enormous diversity of RNA viruses in economic crustaceans.

Crustaceans are important food sources worldwide and possess significant ecological status in the marine ecosystem. However, our understanding of the diversity and evolution of RNA viruses in crustaceans, especially in economic crustaceans, is still limited. Here, 106 batches of economic crustaceans including 13 species were collected from 24 locations in China during 2016-2021. We identified 90 RNA viruses, 69 of which were divergent from the known viruses. Viral transcripts were assigned to 18 different viral families/clades and three unclassified groups. Among the identified viruses, five were double-stranded RNA viruses, 74 were positive-sense single-stranded RNA (+ssRNA) viruses, nine were negative-sense single-stranded RNA (-ssRNA) viruses, and two belonged to an unclassified RNA virus group. Phylogenetic analyses showed that crustacean viruses were often clustered with viruses identified from invertebrates. Remarkably, most crustacean viruses were closely related to those from different host species along the same food chain or ecological aquatic niche. In addition, the genome structures of the newly discovered picornaviruses exhibited remarkable diversity. Our study significantly expands the diversity of viruses in important economic crustaceans and provides essential data for the risk assessment of the pathogens spreading in the global aquaculture industry. IMPORTANCE: The study delves into the largely uncharted territory of RNA viruses in crustaceans, which are not only vital for global food supply but also play a pivotal role in marine ecosystems. Focusing on economic crustaceans, the research uncovers 90 RNA viruses, with 69 being potentially new to science, highlighting the vast unknown viral diversity within these marine organisms. The findings reveal that these viruses are often related to those found in other invertebrates and tend to share close relationships with viruses from species within the same food web or habitat. This suggests that viruses may move between different marine species more frequently than previously thought. The discovery of such a wide variety of viruses, particularly the diverse genome structures of newly identified picornaviruses, is a significant leap forward in understanding the crustacean virology. This knowledge is crucial for managing disease risks in aquaculture and maintaining the balance of marine ecosystems.

Metazoan parasite fauna of grey mullet (Mugilidae) from the coastal waters of Vietnam.

A total of 201 fishes of the family Mugilidae caught off central Vietnam (Nha Trang Bay) were examined for parasites. Eight species of Mugilidae were identified based on cytb and 16S rRNA sequences. Protozoa were not revealed. The metazoan parasites belong to six higher taxa. Myxozoa Grassé, 1970 is represented by six genera, three of which were first found in grey mullets off Vietnam. The specimens of Monogenea Van Beneden, 1858 belong to two genera: Ligophorus Euzet et Suriano, 1977 and Metamicrocotyla Yamaguti, 1953. Twelve species of the former were recorded for the first time both in the coastal waters of Vietnam and in the East Sea. Trematoda Rudolphi, 1808 is represented by the members of the four families, of which only three species, Haplosplanchnus pachysoma (Eysenhardt, 1892), Pseudohaploporus planilizum Atopkin, Besprozvannykh, Ha, Nguyen, Nguyen & Chalenko, 2019 and Spiritestis arabii Nagaty, 1948, were identified. The latter was first recorded in the East Sea. Acanthocephala Rudolphi, 1808 specimens belong to the species Neoechinorhynchus dimorphospinus Amin & Sey, 1996. Crustacea Brünnich, 1772 are represented by five families, of which only the copepod Lernanthropsis mugilii (Shishido, 1898) and the isopod Nerocila orbignyi (Guérin, 1832) were identified. Hirudinea Savigny, 1822 were not determined to genus and species level. A total of 25 species of parasites were found in the Mugilidae fishes examined in Nha Trang Bay. In general, 96 species of parasites are currently documented in these fishes in the East Sea, 55 of which occur off Vietnam. As a result of this study, carried out off central Vietnam, 16 parasite species were recorded in grey mullet for the first time in Vietnamese coastal waters and 15 for the first time in the East Sea. Thus, extending the geographical range of the studies may result in new records of parasites in these fishes.

Sandy beach mole crabs (Decapoda: Hippidae: Emerita): A systematic review of the anthropic impacts, populations density, and conservation strategies.

Emerita species (Scopoli, 1777; Crustacea: Decapoda: Pleocyemata: Anomura: Hippidae), commonly known as mole crabs, are specialized in burying and distributed in the swash zone of the Pacific, Atlantic, and Indian Ocean sandy beaches. This genus has received increasing attention in scientific studies owing to its ecological importance. It is used as a model species to evaluate the ecological state of communities and as a bioindicator of environmental health and quality. Here, we systematically reviewed the literature to understand how Emerita species are affected by anthropogenic activities and gathered records of its population densities. Two independent literature searches were conducted on anthropogenic impact and population density. Anthropogenic impacts were reviewed using 61 articles published between 1974 and 2023. The main anthropogenic impacts were physical changes (n = 19) and toxicity (n = 18). E. analoga, E. talpoida, and E. brasiliensis were the most studied species. Some anthropogenic impacts did not directly damage E. analoga and E. talpoida. E. analoga was more tolerant of certain types of impacts related to physical changes and toxicity. Lower densities of E. brasiliensis were associated with more urbanized beaches. Accessing data on annual cycles, density fluctuations, and population peaks is important to minimize the impacts on populations. In total, 42 articles reported surveys on population density or abundance in nine countries. E. brasiliensis exhibited the highest amount (n = 17) on the beaches in Brazil and Uruguay, followed by E. analoga (n = 13), in Chile and USA. A temporal reduction in the densities was observed for E. brasiliensis, primarily on beaches in Brazil, and E. talpoida. A global standardization of sampling and density and abundance data presentation is necessary for effective long-term assessments of Emerita species. This study synthesizes the impacts that have occurred on Emerita, highlighting potential threats to its conservation.

Parvoviruses of Aquatic Animals.

Family Parvoviridae consists of small, non-enveloped viruses with linear, single-stranded DNA genomes of approximately 4-6 kilobases, subdivided into three subfamilies, Parvovirinae, Densovirinae, and Hamaparvovirinae, and unassigned genus Metalloincertoparvovirus. Parvoviruses of aquatic animals infect crustaceans, mollusks, and finfish. This review describes these parvoviruses, which are highly host-specific and associated with mass morbidity and mortality in both farmed and wild aquatic animals. They include Cherax quadricarinatus densovirus (CqDV) in freshwater crayfish in Queensland, Australia; sea star-associated densovirus (SSaDV) in sunflower sea star on the Northeastern Pacific Coast; Clinch densovirus 1 in freshwater mussels in the Clinch River, Virginia, and Tennessee, USA, in subfamily Densovirinae; hepatopancreatic parvovirus (HPV) and infectious hypodermal and hematopoietic necrosis virus (IHHNV) in farmed shrimp worldwide; Syngnathid ichthamaparvovirus 1 in gulf pipefish in the Gulf of Mexico and parts of South America; tilapia parvovirus (TiPV) in farmed tilapia in China, Thailand, and India, in the subfamily Hamaparvovirinae; and Penaeus monodon metallodensovirus (PmMDV) in Vietnamese P. monodon, in unassigned genus Metalloincertoparvovirus. Also included in the family Parvoviridae are novel parvoviruses detected in both diseased and healthy animals using metagenomic sequencing, such as zander parvovirus from zander in Hungary and salmon parvovirus from sockeye salmon smolts in British Columbia, Canada.

Assessment of health risks associated with heavy metal contamination in selected fish and crustacean species from Temsah Lake, Suez Canal.

Marine pollution caused by heavy metals has emerged as a significant environmental concern, garnering increased attention in recent years. The accumulation of heavy metals in the tissues of marine organisms poses substantial threats to both marine ecosystems and human populations that rely on seafood as a primary food source. Fish and crustaceans are effective biomonitors for assessing heavy metal contamination in aquatic environments. In this study, we determined the concentrations of several heavy metals, including cadmium (Cd), lead (Pb), nickel (Ni), mercury (Hg), and tin (Sn), in four fish species (Mugil cephalus, Mugil capito, L. aurata, and Morone labrax) and five crustacean species (S. rivulatus, Cerastoderma glaucum, Paratapes undulatus, R. decussatus, Callinectes sapidus, and Metapenaeus Stebbingi) from Temsah Lake during both winter and summer seasons. To evaluate the potential ecological and health risks associated with consuming these fish and crustacean species, we calculated the metal pollution index (MPI), weekly intake (EWI), target hazard quotient (THQ), and carcinogenic risk (CR) values. The results revealed a noticeable increase in metal levels during the summer compared to winter in the studied samples. Moreover, the concentration of heavy metals in the muscles of the species generally exceeded those in the liver and gills. The MPI values indicated that Morone labrax exhibited the highest values during winter, while L. aurata showed the highest values during summer. Mugil cephalus demonstrated the lowest MPI values in both seasons. The EWI values for the studied metals were found to be lower than the corresponding tolerable weekly intake (TWI) values. Additionally, under average exposure conditions, the THQ and HI data were generally below one for most study species in the area. The calculated CR values for investigated metals in the studied species indicated acceptable carcinogenic risk levels. Therefore, this suggests that consuming studied species within Temsah lake does not present any potential health hazards for consumers.

Application of the Biological Toxicity Effect Ratio (BER) Method for Advancing Water Quality Criteria Derivation.

Water quality criteria (WQC) serve as a scientific foundation for pollutant risk assessment and control in aquatic ecosystems. The development of regionally differentiated WQC tailored to specific regional characteristics has become an emerging trend. However, the current WQC is constrained by a lack of regional species toxicity data. To address these limitations, this study proposes the biological toxicity effect ratio (BER) method, which indirectly reflects the toxicity sensitivity of the overall aquatic ecosystem through the toxicity information on a limited number of species, enabling rapid WQC prediction. Using the established WQC in China and the USA as a case study, we combined mathematical derivation and data validation to evaluate the BER method. Among various species-taxon groups of freshwater organisms, planktonic crustaceans demonstrated the highest predictive accuracy. Our analysis further revealed that species toxicity sensitivity and regional variability jointly influence the prediction accuracy. Regardless of the evaluation indexes, planktonic crustaceans emerged as the most suitable species-taxon group for the BER method. Additionally, the BER method is particularly applicable to pollutants with conserved mechanisms across species. This study systematically explores the feasibility of using the BER method and offers new insights for deriving regionally differentiated WQC.

Holistic assessment of dimethoate toxicity in Carcinus aestuarii's muscle tissues.

Dimethoate (DMT) is one of the most harmful and commonly used organophosphate pesticides in agricultural lands to control different groups of parasitic insects. However, this pesticide is considered a dangerous pollutant for aquatic organisms following its infiltration in coastal ecosystems through leaching. Yet, our investigation aimed to gain new insights into the toxicity mechanism of DMT in the muscles of the green crab Carcinus aestuarii, regarding oxidative stress, neurotransmission impairment, histological aspects, and changes in lipid composition, assessed for the first time on the green crab's muscle. Specimens of C. aestuarii were exposed to 50, 100, and 200 µg DMT L-1 for 24 h. Compared to the negative control group, the higher the DMT concentration, the lower the saturated fatty acids (SFA), and the higher the monounsaturated fatty acids (MUFA). The significant increase in polyunsaturated fatty acid n-6 (PUFA n-6) was related to the high release, mainly, of linoleic acid (LA, C18: 2n6) and arachidonic acid (ARA, C20: 4n6) levels. Biochemical biomarkers showed that DMT exposure promoted oxidative stress, highlighted by increased levels of hydrogen peroxide (H2O2), malondialdehyde (MDA), advanced oxidation protein product levels (AOPP), and protein carbonyl (PCO). Furthermore, the antioxidant defense system was activated, as demonstrated by the significant changes in the enzymatic activity of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), and reduced glutathione (GSH) levels associated with an adaptation process of C. aestuarii to cope with the DMT exposure. This pesticide significantly impairs the neurotransmission process, as evidenced by the inhibition of acetylcholinesterase (AChE) activity. Finally, several histopathological changes were revealed in DMT-treated crabs, including vacuolation, and muscle bundle loss.This research offered new insights into the toxic mechanism of DMT, pointing to the usefulness of fatty acid (FA) composition as a sensitive biomarker in littoral crabs.

Diversely evolved xibalbin variants from remipede venom inhibit potassium channels and activate PKA-II and Erk1/2 signaling.

BACKGROUND: The identification of novel toxins from overlooked and taxonomically exceptional species bears potential for various pharmacological applications. The remipede Xibalbanus tulumensis, an underwater cave-dwelling crustacean, is the only crustacean for which a venom system has been described. Its venom contains several xibalbin peptides that have an inhibitor cysteine knot (ICK) scaffold. RESULTS: Our screenings revealed that all tested xibalbin variants particularly inhibit potassium channels. Xib1 and xib13 with their eight-cysteine domain similar to spider knottins also inhibit voltage-gated sodium channels. No activity was noted on calcium channels. Expanding the functional testing, we demonstrate that xib1 and xib13 increase PKA-II and Erk1/2 sensitization signaling in nociceptive neurons, which may initiate pain sensitization. Our phylogenetic analysis suggests that xib13 either originates from the common ancestor of pancrustaceans or earlier while xib1 is more restricted to remipedes. The ten-cysteine scaffolded xib2 emerged from xib1, a result that is supported by our phylogenetic and machine learning-based analyses. CONCLUSIONS: Our functional characterization of synthesized variants of xib1, xib2, and xib13 elucidates their potential as inhibitors of potassium channels in mammalian systems. The specific interaction of xib2 with Kv1.6 channels, which are relevant to treating variants of epilepsy, shows potential for further studies. At higher concentrations, xib1 and xib13 activate the kinases PKA-II and ERK1/2 in mammalian sensory neurons, suggesting pain sensitization and potential applications related to pain research and therapy. While tested insect channels suggest that all probably act as neurotoxins, the biological function of xib1, xib2, and xib13 requires further elucidation. A novel finding on their evolutionary origin is the apparent emergence of X. tulumensis-specific xib2 from xib1. Our study is an important cornerstone for future studies to untangle the origin and function of these enigmatic proteins as important components of remipede but also other pancrustacean and arthropod venoms.

Effects of glyphosate on sperm quality of the estuarine crab Neohelice granulata, under the "One Health" perspective.

This study was aimed at evaluating the effects of the commercial formulation Roundup Ultramax® on sperm mortality and viability, in terms of the capacity of spermatozoids (spz) to trigger the acrosome reaction (AR), using the estuarine crab Neohelice granulata as a model. To this, an in vivo assay comprising 100 days duration was carried out, on a control group and two groups exposed to the formulation (0.01 mg/L and 0.2 mg/L of glyphosate) under controlled conditions of photoperiod, feeding, and temperature. At the end of the assay, the right vas deferens (VD, proximal and middle portion) was dissected, and after homogenizing it in calcium-free saline solution, the acrosome reaction was induced in the phase containing the spz. In each treatment, the percentage of spz with total and partial AR was calculated, as well as that of dead spz. Compared to the control, crabs exposed to the herbicide showed a significant decrease in spz with full AR, together with an increase in the percentage of spz with partial AR. Furthermore, spz mortality was significantly higher in both glyphosate concentrations compared to the control, in a concentration-dependent manner. On the other hand, abnormal spermatophores, showing expanded walls and coalescence, were observed in a significant percentage in the left VD of the groups treated with the herbicide. The results obtained are compared with those from other studies on several invertebrate and vertebrate species that found inhibition of the AR and abnormal sperm, together with inhibition of spermatogenesis, endocrine disruption, and reduced sperm motility by effect of pure glyphosate and/or different glyphosate formulations. In summary, the available evidence highlights the possible impact of glyphosate on sperm quality, in a wide variety of species.

The mucosal immunity in crustaceans: Inferences from other species.

Crustaceans such as shrimps and crabs, hold significant ecological significance and substantial economic value within marine ecosystems. However, their susceptibility to disease outbreaks and pathogenic infections has posed major challenges to production in recent decades. As invertebrate, crustaceans primarily rely on their innate immune system for defense, lacking the adaptive immune system found in vertebrates. Mucosal immunity, acting as the frontline defense against a myriad of pathogenic microorganisms, is a crucial aspect of their immune repertoire. This review synthesizes insights from comparative immunology, highlighting parallels between mucosal immunity in vertebrates and innate immune mechanisms in invertebrates. Despite lacking classical adaptive immunity, invertebrates, including crustaceans, exhibit immune memory and rely on inherent "innate immunity factors" to combat invading pathogens. Drawing on parallels from mammalian and piscine systems, this paper meticulously explores the complex role of mucosal immunity in regulating immune responses in crustaceans. Through the extrapolation from well-studied models like mammals and fish, this review infers the potential mechanisms of mucosal immunity in crustaceans and provides insights for research on mucosal immunity in crustaceans.

Back to the future: Forgotten protocols for optimizing the isolation of arthropod haemocytes.

Consideration is given to previous and more recent protocols for harvesting arthropod haemocytes from Galleria, Drosophila, mosquitoes, Limulus and crustaceans. The optimal harvesting of these cells is essential for meaningful studies of invertebrate immunity in vitro. The results of such experiments, however, have often been flawed due to a lack of understanding of the fragile nature of arthropod haemocytes on exposure to bacterial lipopolysaccharides, resulting in the aggregation and loss of cell types during haemolymph clotting. This article emphasizes that although there are similarities between mammalian neutrophils and arthropod haemocytes, the protocols required for the successful harvesting of these cells vary significantly. The various stages for the successful harvesting of arthropod haemocytes are described in detail and should provide invaluable advice to those requiring both high cell viability and recovery of the different cell types for subsequent experimentation.

Quantification of Allergic Crustacean Tropomyosin Using Shared Signature Peptides in Processed Foods with a Mass Spectrometry-Based Proteomic Strategy.

Crustacean shellfish are major allergens in East Asia. In the present study, a major allergic protein in crustaceans, tropomyosin, was detected accurately using multiple reaction monitoring mode-based mass spectrometry, with shared signature peptides identified through proteomic analysis. The peptides were deliberately screened through thermal stability and enzymatic digestion efficiency to improve the suitability and accuracy of the developed method. Finally, the proposed method demonstrated a linear range of 0.15 to 30 mgTM/kgfood (R2 > 0.99), with a limit of detection of 0.15 mgTM/kg food and a limit of quantification of 0.5mgTM/kgfood and successfully applied to commercially processed foods, such as potato chips, biscuits, surimi, and hot pot seasonings, which evidenced the applicability of proteomics-based methodology for food allergen analysis.

Effects of high temperature and LPS injections on the hemocytes of the crab Neohelice granulata.

Temperature fluctuations, particularly elevated temperatures, can significantly affect immune responses. These fluctuations can influence the immune system and alter its response to infection signals, such as lipopolysaccharide (LPS). Therefore, this study was designed to investigate how high temperatures and LPS injections collectively influence the immune system of the crab Neohelice granulata. Two groups were exposed to 20 °C (control) or 33 °C for four days. Subsequently, half were injected with 10 µL of physiological crustacean (PS), while the rest received 10 µL of LPS [0.1 mg.kg-1]. After 30 min, the hemolymph samples were collected. Hemocytes were then isolated and assessed for various parameters using flow cytometry, including cell integrity, DNA fragmentation, total hemocyte count (THC), differential hemocyte count (DHC), reactive oxygen species (ROS) level, lipid peroxidation (LPO), and phagocytosis. Results showed lower cell viability at 20 °C, with more DNA damage in the same LPS-injected animals. There was no significant difference in THC, but DHC indicated a decrease in hyaline cells (HC) at 20 °C following LPS administration. In granular cells (GC), an increase was observed after both PS and LPS were injected at the same temperature. In semi-granular cells (SGC), there was a decrease at 20 °C with the injection of LPS, while at a temperature of 33 °C, the SGC there was a decrease only in SGC injected with LPS. Crabs injected with PS and LPS at 20 °C exhibited higher levels of ROS in GC and SGC, while at 33 °C, the increase was observed only in GC and SGC cells injected with LPS. A significant increase in LPO was observed only in SGC cells injected with PS and LPS at 20 °C and 33 °C. Phagocytosis decreased in animals at 20 °C with both injections and exposed to 33 °C only in those injected with LPS. These results suggest that elevated temperatures induce changes in immune system parameters and attenuate the immune responses triggered by LPS.

The complete mitochondrial genome of the sand bubbler crab Scopimera longidactyla Shen 1932 (Decapoda: Dotillidae) and its phylogenetic analysis.

The sand bubbler crab, Scopimera longidactyla Shen, 1932 (Arthropoda: Malacostraca: Decapoda: Thoracotremata: Dotillidae), is commonly found along tropical and subtropical sandy shores of China, Korea, and Taiwan. Ecologically, it plays an important role in the productivity of sandy shores through their feeding and burrowing activities. In this study, the first complete mitochondrial genome (mitogenome) of S. longidactyla was analyzed using next-generation sequencer. Its mitogenome, circular in structure, spans 15,965 bp with a GC content of 29.97%, consisting of 13 protein-coding genes, two ribosomal RNA genes, 22 transfer RNA genes, and one putative control region. Its mitogenome arrangement and composition are identical to its two congeners, S. globosa and S. intermedia. Phylogenetic analysis fully supports for the monophyly of the genus Scopimera and the sister relationship between S. longidactyla and S. globosa. The complete mitogenome of S. longidactyla and its phylogenetic implications will provide valuable insights for further studies in phylogenetic and evolutionary biology.

Chitin quantitation (as glucosamine) in food raw materials by HPLC-C18-DAD with off-line derivatization.

This HPLC method is suitable for chitin quantitation (reported as glucosamine) in food raw materials like insects (mealworm larvae, crickets), shrimps, mushrooms and fungi in a research (non-routine) laboratory using a C18 column with HPLC system <600 bar with UV detection capability (at 265 nm). To remove interferences, the sample is defatted (Soxhlet) and deproteinized (by alkali) prior to acid hydrolysis in 6 M HCl. A five-point linear calibration (5-100 µg/mL) is used. The use of fluorescence detection (λex = 260 nm, λem = 350 nm) is also possible with this method [1].•18 min HPLC run time•LOD = 0.05 µg/mL and LOQ = 5 µg/mL.

Statocyst Ultrastructure in the Norwegian Lobster (Nephrops norvegicus).

Statocyst anatomy and fine morphology in Norwegian lobster (Nephrops norvegicus) are studied for the first time using scanning and transmission electron microscopy. N. norvegicus exhibits sensory setae projecting from the statocyst inner cavity floor into a mass of sand granules (statoconia) embedded in a gelatinous substance. The setae are distributed in four areas: a curved field made up of an inner single row and an outer double row that run on a circle around the medial and lateral rim of the central depression, a small setal field in the posterior part, a large setal field, opposite to the small field, and a short row, running internally and lying parallel to the inner single row, next to the small setal field. A study of the fine morphology of the statocyst sensory setae shows that the structure of the setae in the different areas is similar, with a bulb (the proximal portion of the sensillum), a setal shaft, a tooth (the smooth portion of the bulb), a fulcrum (a transverse fold), and filamentous hairs. The hair cells are firmly implanted within the cuticular layer. Although the type of innervation of the statocyst was not determined in the present study, the close taxonomic position of the lobster to that of the crayfish and crab would suggest that the setae in N. norvegicus are pure mechanoreceptors rather than sensory cells.

Environmental Transmission of Symbionts in the Mangrove Crabs Aratus pisonii and Minuca rapax: Acquisition of the Bacterial Community through Larval Development to Juvenile Stage.

Aratus pisonii and Minuca rapax are two brachyuran crabs living with bacterial ectosymbionts located on gill lamellae. One previous study has shown that several rod-shaped bacterial morphotypes are present and the community is dominated by Alphaproteobacteria and Bacteroidota. This study aims to identify the mode of transmission of the symbionts to the new host generations and to identify the bacterial community colonizing the gills of juveniles. We tested for the presence of bacteria using PCR with universal primers targeting the 16S rRNA encoding gene from gonads, eggs, and different larval stages either obtained in laboratory conditions or from the field. The presence of bacteria on juvenile gills was also characterized by scanning electron microscopy, and subsequently identified by metabarcoding analysis. Gonads, eggs, and larvae were negative to PCR tests, suggesting that bacteria are not present at these stages in significant densities. On the other hand, juveniles of both species display three rod-shaped bacterial morphotypes on gill lamellae, and sequencing revealed that the community is dominated by Bacteroidota and Alphaproteobacteria on A. pisonii juveniles, and by Alphaprotobacteria, Bacteroidota, and Acidimicrobia on M. rapax juveniles. Despite the fact that juveniles of both species co-occur in the same biotope, no shared bacterial phylotype was identified. However, some of the most abundant bacteria present in adults are also present in juveniles of the same species, suggesting that juvenile-associated communities resemble those of adults. Because some of these bacteria were also found in crab burrow water, we hypothesize that the bacterial community is established gradually during the life of the crab starting from the megalopa stage and involves epibiosis-competent bacteria that occur in the environment.

Effects of Temperature and Salinity on Perfluorooctane Sulfonate (PFOS) Toxicity in Larval Estuarine Organisms.

Perfluorooctane sulfonate (PFOS) is a persistent contaminant that has been found globally within the environment. Key data gaps exist in the toxicity of PFOS to marine organisms, especially estuarine species that are crucial to the food web: fish, shrimp, and mollusks. This study developed toxicity thresholds for larval estuarine species, including grass shrimp (Palaemon pugio), sheepshead minnows (Cyprinodon variegatus), mysids (Americamysis bahia), and Eastern mud snails (Tritia obsoleta). Multiple abiotic stressors (salinity and temperature) were included as variables in testing the toxicity of PFOS. Acute 96 h toxicity testing under standard test conditions of 25 °C and 20 ppt seawater yielded LC50 values of 0.919 mg/L for C. variegatus, 1.375 mg/L for A. bahia, 1.559 mg/L for T. obsoleta, and 2.011 mg/L for P. pugio. The effects of increased temperature (32 °C) and decreased salinity (10 ppt) varied with test species. PFOS toxicity for the sheepshead minnows increased with temperature but was not altered by decreased salinity. For grass shrimp and mud snails, PFOS toxicity was greater under lower salinity. The combination of higher temperature and lower salinity was observed to lower the toxicity thresholds for all species. These data demonstrate that expanding toxicity testing to include a wider range of parameters will improve the environmental risk assessment of chemical contaminants, especially for species inhabiting dynamic estuarine ecosystems.

Chitin quantitation (as glucosamine) in food raw materials by spectrophotometry.

Chitin is a water insoluble nitrogen-containing polysaccharide made from N-acetyl-D-glucosamine containing ß-(1→4)-linkages. In food, chitin is considered as a source of fiber with prebiotic properties to gut microflora. Chitin content varies widely in nature from 1% (yeasts) up to 64% (butterfly cuticles) and is mostly found in filamentous or mushroom forming fungi, insects and crustaceans. This spectrophotometric method is suitable for chitin quantitation (reported as glucosamine) in food raw materials like insects (mealworm larvae, crickets), shrimps, mushrooms and fungi in a research (non-routine) laboratory. To remove interferences, the sample is defatted (Soxhlet) prior to acid hydrolysis in 6 M HCl. The color complex is developed after the addition of Katano's reagent (a mix of 0.05 mol/L sodium metasilicate, 0.6 mol/L sodium molybdate, 30% dimethyl sulfoxide and 1.42 mol/L acetic acid) at 70 °C for 30 min and measured at 750 nm against blank. A five-point linear calibration (5-100 µg/mL) is used. Limit of detection is 3 µg GLCN/mL. The correlation (R2) with an HPLC method for chitin analysis is at least 0.93.•a reliable alternative to an HPLC method•does not require expensive equipment•deproteination by alkali is not necessary for most matrices - saves about 30% of time.