Improving photovoltaic cell parameter calculations through a puffer fish inspired optimization technique.

The precise estimation of solar PV cell parameters has become increasingly important as solar energy deployment expands. Due to the intricate and nonlinear characteristics of solar PV cells, meta-heuristic algorithms show greater promise than traditional ones for parameter estimation. This study utilizes the Puffer Fish (PF) meta-heuristic optimization method, inspired by male puffer fish's circular structures, to estimate parameters of a modified four-diode PV cell. The PF algorithm's performance is assessed against ten benchmark test functions, with results presented as mean and standard deviation for validation. Comparative analysis with Particle Swarm Optimization (PSO), Grey Wolf Optimization (GWO), Rat Search Algorithm (RAT), Heap Based Optimizer (HBO), and Cuckoo Search (CS) algorithms highlights PF's superior performance, achieving optimal solutions with minimal error of 7.8947E-08. Statistical tests, including Friedman Ranking (1st) and Wilcoxon's rank sum (3.8108E-07), confirm PF's superiority. The circular structures of male puffer fish serve as an effective model for optimization algorithms, enhancing parameter estimation. Benchmark tests and statistical analysis consistently underscore PF's superiority over other meta-heuristic algorithms. Future research should explore PF's potential applications in solar energy and beyond.

Effect of plasma-activated lactic acid on microbiota composition and quality of puffer fish (Takifugu obscurus) fillets during chilled storage.

Fresh puffer fish (Takifugu obscurus) are susceptible to microbial contamination and have a very short shelf-life of chilled storage. Hence, this study aimed to evaluate the effects of plasma-activated lactic acid (PALA) on microbiota composition and quality attributes of puffer fish fillets during chilled storage. The results showed that PALA treatment effectively reduced the growth of bacteria and attenuated changes in physicochemical indicators (total volatile basic nitrogen, pH value, K value, and biogenic amines) of puffer fish fillets. Additionally, insignificant changes were observed in lipid oxidation during the first 8 days (p > 0.05). Illumina-MiSeq high-throughput sequencing revealed that PALA effectively inhibited the growth of Pseudomonas in puffer fish fillets and maintained the diverse characteristics of the microbial community. In combination with sensory analysis, PALA extended the shelf life of puffer fish fillets for 4 days, suggesting that PALA could be considered a potential fish fillet preservation method.

Spikebot: A Multigait Tensegrity Robot with Linearly Extending Struts.

Numerous recent research efforts have leveraged networks of rigid struts and flexible cables, called tensegrity structures, to create highly resilient and packable mobile robots. However, the locomotion of existing tensegrity robots is limited in terms of both speed and number of distinct locomotion modes, restricting the environments that a robot is capable of exploring. In this study, we present a tensegrity robot inspired by the volumetric expansion of Tetraodontidae. The robot, referred to herein as Spikebot, employs pneumatically actuated rigid struts to expand its global structure and produce diverse gaits. Spikebot is composed of linear actuators that dually serve as rigid struts linked by elastic cables for stability. The linearly actuating struts can selectively protrude to initiate thrust- and instability-driven locomotion primitives. Such motion primitives allow Spikebot to reliably locomote, achieving rolling, lifting, and jumping. To highlight Spikebot's potential for robotic exploration, we demonstrate how it achieves multi-dimensional locomotion over varied terrestrial conditions.

Differential blood cells associated with parasitism in the wild puffer fish Lagocephalus laevigatus (Tetraodontiformes) of the Campeche Coast, southern Mexico.

In aquaculture conditions, severe parasitic infections cause negative impacts on fish health and economic losses. The parasite load has been associated with anemia, which reduces the number of erythrocytes in fish. Therefore, the evaluation of hematological parameters as a feasible tool for diagnosing and monitoring fish health allows us to determine the indirect effect of parasites on the health status of fish. Our aim was to evaluate changes in the blood cell parameters of Lagocephalus laevigatus associated with parasitism. A total of 99 puffer fish were collected from the coast of Seybaplaya, Campeche. Each fish had 20 µl of peripheral blood drawn, and blood smears were performed in triplicate. The smears were stained with Giemsa stain, and a quantitative analysis of blood cells (erythrocytes, leukocytes, and monocytes) was obtained with an optical microscope at 100 ×. The parasites recovered from each fish were fixed and identified, and the infection parameters were calculated. Through generalized additive model analysis (GAMLSS), we observed that the infection intensity of puffer fish influenced changes in hematological parameters, principally in erythrocytes, neutrophils, thrombocytes, the total fish length, and the condition factor of the fish. In conclusion, this is the first study that provides baseline data on the hematological parameter variations in uninfected and infected L. laevigatus, the tropical wild puffer fish, as well as the possible effects on fish health. It is necessary to establish reference hematological patterns in wild populations for diagnosis and timely management with emphasis on aquaculture fish.

Histological and ultrastructural characterization of the dorso-ventral skin of the juvenile and the adult starry puffer fish (Arothron stellatus, Anonymous 1798).

BACKGROUND: The starry puffer fish (Arothron stellatus, Anonymous, 1798) is a poisonous tetradontidae fish inhabiting the Red sea. The skin constitutes an important defense against any external effects. The study aims to characterize the dorso-ventral skin of the juvenile and the adult starry puffer fish using light and scanning electron microscopies. Twenty specimens of juvenile and adult fresh fishes were used. RESULTS: The scanning electron microarchitecture of the skin of the juvenile and adult fish showed delicate irregular-shaped protrusions, and well-defined bricks-like elevations on the dorsal side and interrupted folds as well as irregular-shaped protrusions on the ventral side. In adult fish, the patterned microridges of the superficial and deep epithelial cells (keratinocytes) were larger and well-defined in the dorsal skin than in the ventral side, the contrary was seen in the juvenile fish. The microridges were arranged in a fingerprint or honeycomb patterns. The openings of the mucous cells were more numerous in the dorsal skin in both age stages but more noticeable in adult. Furthermore, the sensory cells were more dominant in the juveniles than the adults. The odontic spines were only seen in adult. Histologically, few taste buds were observed in the epidermis of the dorsal skin surface of the adult fish. Both mucous and club cells were embedded in the epidermis of the juvenile and adult fish with different shapes and sizes. Melanophores were observed at the dorsal skin of both juvenile and adult fishes while fewer numbers were noticed at the ventral surfaces. Several dermal bony plates with different shapes and sizes were demonstrated in the skin of both adult and juvenile fishes. CONCLUSION: The structural variations of skin of the juvenile and adult fishes may reflect the various environmental difficulties that they confront.

Evaluation of Toxicity Equivalency Factors of Tetrodotoxin Analogues with a Neuro-2a Cell-Based Assay and Application to Puffer Fish from Greece.

Tetrodotoxin (TTX) is a potent marine neurotoxin involved in poisoning cases, especially through the consumption of puffer fish. Knowledge of the toxicity equivalency factors (TEFs) of TTX analogues is crucial in monitoring programs to estimate the toxicity of samples analyzed with instrumental analysis methods. In this work, TTX analogues were isolated from the liver of a Lagocephalus sceleratus individual caught on South Crete coasts. A cell-based assay (CBA) for TTXs was optimized and applied to the establishment of the TEFs of 5,11-dideoxyTTX, 11-norTTX-6(S)-ol, 11-deoxyTTX and 5,6,11-trideoxyTTX. Results showed that all TTX analogues were less toxic than the parent TTX, their TEFs being in the range of 0.75-0.011. Then, different tissues of three Lagocephalus sceleratus individuals were analyzed with CBA and liquid chromatography-tandem mass spectrometry (LC-MS/MS). The obtained TEFs were applied to the TTX analogues' concentrations obtained by LC-MS/MS analysis, providing an indication of the overall toxicity of the sample. Information about the TEFs of TTX analogues is valuable for food safety control, allowing the estimation of the risk of fish products to consumers.

A Case of Puffer Fish Poisoning from United Arab Emirates.

Puffer fish (Lagocephalus sceleratus) is a well-known source of deadly food poisoning due to a neurotoxin, called tetrodotoxin. Cases of tetrodotoxin poisoning are common along the coasts of East Asian countries but rare in the Arabian Gulf region. Here, we report a case of a 19-year-old man who presented with signs and symptoms suggestive of puffer fish poisoning. Dietary history was the key to diagnosis though laboratory investigations and imaging yielded normal results. Early diagnosis and proper supportive management are essential for survival.

Development of a Proton-Enhanced ESI UPLC-MS/MS Method for the Determination of Tetrodotoxin.

Tetrodotoxin (TTX) is a kind of low-molecular-weight non-protein neurotoxin. It is one of the most potent neurotoxins found in nature, and it is found in puffer fish and various marine biota. The low sensitivity of previous analytical methods limited their application in puffer fish organ samples. This study established a method for the accurate and fast determination of TTX by reversed ultra-performance liquid chromatography coupled with proton-enhanced electron spray ionization-tandem mass spectrometry. The method yields good peak shapes, high sensitivity and low coeluted interferences. The method was successfully applied to determine TTX in puffer fish tissue samples of about 0.2 g.

Real-time qPCR for the detection of puffer fish components from Lagocephalus in food: L. inermis, L. lagocephalus, L. gloveri, L. lunaris, and L. spadiceus.

Puffer fish is a type of precious high-end aquatic product, is widely popular in Asia, especially in China and Japan, even though it naturally harbors a neurotoxin known as tetrodotoxin (TTX) that is poisonous to humans and causes food poisoning. With the increasing trade demand, which frequently exceeds existing supply capacities, fostering fraudulent practices, such as adulteration of processed products with non-certified farmed wild puffer fish species. To determine the authenticity of puffer fish processed food, we developed a real-time qPCR method to detect five common puffer fish species in aquatic products: Lagocephalus inermis, Lagocephalus lagocephalus, Lagocephalus gloveri, Lagocephalus lunaris, and Lagocephalus spadiceus. The specificity, cross-reactivity, detection limit, efficiency, and robustness of the primers and probes created for five species of puffer fish using TaqMan technology have been determined. No cross-reactivity was detected in the DNA of non-target sample materials, and no false-positive signal was detected; the aquatic products containing 0.1% of a small amount of wild puffer fish materials without certification can be reliably tracked; the statistical p-value for each method's Ct value was greater than 0.05. The developed qPCR method was sensitive, highly specific, robust, and reproducibility, which could be used to validate the authenticity of wild puffer fish in aquatic products sold for commercial purposes.

[Tetrodotoxin Intoxications in Nagasaki, Japan: Symptoms of Patients, Tetrodotoxin Levels in Leftover Food and Clinical Urine and Serum Samples].

Incidents of food poisoning associated with tetrodotoxin (TTX) contamination occur every year throughout Japan. Here, we determined TTX levels in leftover foods and serum and urine samples from eight food poisoning incidents associated with TTX contamination in Nagasaki, Japan, from 2011 to 2017.Seven food samples associated with four of these food poisoning incidents were classified as weakly toxic (four samples), moderate-to-strongly toxic (two samples), and strongly toxic (one sample).In comparison with previous reports, TTX was detected at harmful levels in the urine samples, but the grade of poisoning symptoms varied.The patients' time to maximum serum TTX levels (Tmax) was estimated to be 12-24 h after ingestion of TTX-containing foods. Additionally, serum TTX levels of 19.5 ng/mL or higher within 24 h after ingestion indicated Grade 3 poisoning associated with respiratory abnormalities.These conditions were considered indicators of severe symptoms, while TTX levels of 1-3 ng/mL relate to the onset and disappearance of symptoms. A negative correlation was found between the logarithm of serum TTX concentration and the time after ingestion for two patients, indicating that the TTX serum levels decreased logarithmically. Furthermore, the TTX serum half-lives (T1/2) were 17.5 and 23.7 h.The results of this study enhance our understanding of TTX food safety and contribute to TTX risk assessment.

Kisspeptin Exhibits Stimulatory Effects on Expression of the Genes for Kisspeptin Receptor, GnRH1 and GTH Subunits in a Gonadal Stage-Dependent Manner in the Grass Puffer, a Semilunar-Synchronized Spawner.

Kisspeptin has an important role in the regulation of reproduction by directly stimulating the secretion of gonadotropin-releasing hormone (GnRH) in mammals. In non-mammalian vertebrates, there are multiple kisspeptins (Kiss1 and Kiss2) and kisspeptin receptor types, and the two kisspeptins in teleosts have different effects depending on fish species and reproductive stages, serving reproductive and non-reproductive functions. In the grass puffer, Takifugu alboplumbeus, which has only a single pair of kiss2 and kissr2, both genes display seasonal, diurnal, and circadian oscillations in expression in association with the periodic changes in reproductive functions. To elucidate the role of kisspeptin in this species, homologous kisspeptin peptide (gpKiss2) was administered at different reproductive stages (immature, mature and regressed) and the expression levels of the genes that constitute hypothalamo-pituitary-gonadal axis were examined in male grass puffer. gpKiss2 significantly elevated the expression levels of kissr2 and gnrh1 in the brain and kissr2, fshb and lhb in the pituitary of the immature and mature fish. No noticeable effect was observed for kiss2, gnih, gnihr, gnrh2 and gnrh3 in the brain and gpa in the pituitary. In the regressed fish, gpKiss2 was ineffective in stimulating the expression of the gnrh1 and GTH subunit genes, while it stimulated and downregulated the kissr2 expression in the brain and pituitary, respectively. The present results indicate that Kiss2 has a stimulatory role in the expression of GnRH1/GTH subunit genes by upregulating the kissr2 expression in the brain and pituitary at both immature and mature stages, but this role is mostly ineffective at regressed stage in the grass puffer.

Metabolism and pharmacokinetics of mebendazole in Japanese pufferfish (Takifugu rubripes).

As a typical and broad-spectrum benzimidazole, mebendazole (MBZ) has long been used in human and veterinary medicine to treat parasitic infestations, and is widely employed in the aquaculture of Japanese pufferfish (Takifugu rubripes). However, there have been no studies examining the pharmacokinetic characteristics of MBZ in Japanese pufferfish. Furthermore, the presence of MBZ and its metabolites in animal-derived raw food represents a notable safety concern. Here, we investigated the metabolism of MBZ using a UPLC-Q-TOF system. Additionally, we evaluated the pharmacokinetics of MBZ and two metabolites, 2-amino-5(6)-benzoylbenzimidazole (MBZ-NH2) and 5-hydroxymebendazole (MBZ-OH), in Japanese pufferfish following intramuscular injection of 20 mg/kg MBZ. We detected three metabolites of MBZ (M1-M3), among which, 2-amino-5(6)-(a-hydroxybenzyl) benzimidazole (M3) was detected in an aquatic animal for the first time. The plasma dispositions of MBZ, MBZ-NH2, and MBZ-OH were characterized by low plasma clearance, medium distribution volume, and long terminal half-life. Moreover, these compounds were widely distributed in the muscle, from which they were rapidly cleared. The pharmacokinetics and metabolism of mebendazole in Japanese pufferfish are described for the first time in this study. Our findings provide a basis for the rational application of MBZ in Japanese pufferfish farming and contribute to our understanding of the metabolism of MBZ in cultured fish.

Morphology and molecular phylogeny of Heterobothrium lamothei (Monogenea: Diclidophoridae), a gill parasite infecting the tiger puffer fish Lagocephalus sceleratus (Tetraodontidae) / Morfologia e filogenia molecular de Heterobothrium lamothei (Monogenea: Diclidophoridae), um parasita das guelras que infecta o peixe tigre pufferLagocephalus sceleratus (Tetraodontidae)

Members of the family Diclidophoridae are potentially dangerous species for the puffer fish aquaculture worldwide. They are parasitic polyopisthocotyleans, with a posterior haptor equipped with clamps for attachment to the host's surface, allowing the worm to resist the flow of water to maintain its position on gills. The anterior body of the worm is deformable, allows the worm to feed on blood sucked from fish gills. The present study is the first description of a Heterobothrium species from the gills of the tiger puffer Lagocephalus sceleratus (Tetraodontidae) from the coasts of the Arabian Gulf at Jubail, Saudi Arabia morphologically by light microscopy as well as by molecular analysis of the parasite partial 28S rRNA through multiple sequence alignments and phylogeny by maximum likelihood analysis which is provided for the first time for the described species. Seventeen tiger puffer fish were captured alive from marine water off Saudi Arabia; gills were separated and further examined for parasitic infection. Nine fish were found infected with a monogenean parasite which was robust, equipped by two buccal organs at the tapered anterior end; the posterior haptor was rectangular with four symmetrically arranged clamps, with no isthmus. Marginal hooks absent. Ovary elongated, U-shaped, testes numerous, irregularly shaped and extended from the posterior part of the ovary to the anterior margin of the haptor. Copulatory organ muscular, as a spherical cup armed with 12 to 15 genital hooks. The molecular analysis of the parasite 28s rRNA and phylogeny revealed a percentage of identities between 87.47-89.09%, with Diclidophoridae species within the monophyletic clade of Mazocraeidea where a maximum percentage of 89.09% were obtained for the morphologically different sister taxon H. okamotoi. The results obtained from molecular analysis are consistent with the conclusions drawn from morphological classification where that the parasite recorded was morphologically similar to H. lamothei which was not characterized by molecular analysis before. The recovered sequences were deposited into the GeneBank under accession number MT322610.(AU)

Os membros da família Diclidophoridae são espécies potencialmente perigosas para a aquicultura de peixes puffer em todo o mundo. Eles são parasitas poliopisthocotyleans, com uma hélice posterior equipada com pinças para fixação na superfície do hospedeiro, permitindo que o verme resista ao fluxo de água para manter sua posição nas brânquias. O corpo anterior do verme é deformável, e permite que o verme se alimente de sangue sugado das guelras dos peixes. O presente estudo é a primeira descrição de uma espécie Heterobothrium das guelras do tigre Lagocephalus sceleratus (Tetraodontidae) das costas do Golfo Arábico em Jubail, Arábia Saudita, usando morfologia por microscopia leve, bem como análise molecular do rRNA parcial do parasita 28S através de alinhamentos de sequências múltiplas e filogenia por análise de máxima verossimilhança que é fornecida pela primeira vez para as espécies descritas. Dezessete peixes tigre puffer foram capturados vivos da água marinha da Arábia Saudita; as brânquias foram separadas e mais tarde examinadas para detecção de infecção parasitária. Nove peixes foram encontrados infectados por um parasita monogênio robusto, equipado por dois órgãos bucais na extremidade anterior afilada; o hortetor posterior era retangular com quatro pinças dispostas simetricamente, sem istmo. Ausência de ganchos marginais. Ovário alongado, em forma de U, testículos numerosos, de forma irregular e estendido desde a parte posterior do ovário até a margem anterior do hortelino. Órgão copulatório muscular, como um copo esférico armado com 12 a 15 ganchos genitais. A análise molecular do parasita 28s rRNA e filogenia revelou uma porcentagem de identidades entre 87,47-89,09%, com espécies Diclidophoridae dentro do clade monofilético de Mazocraeidea onde uma porcentagem máxima de 89,09% foi obtida para o táxon-irmão morfologicamente diferente H. okamotoi. Os resultados obtidos da análise molecular são consistentes com as conclusões tiradas da classificação morfológica onde o parasita registrado era morfologicamente semelhante ao H. lamothei que não era caracterizado pela análise molecular antes. As sequências recuperadas foram depositadas no GeneBank sob o número de acesso MT322610.(AU)

Expression dynamics of the genes for the hypothalamo-pituitary-gonadal axis in tiger puffer (Takifugu rubripes) at different reproductive stages.

Tiger puffer, Takifugu rubripes, a commercially important long-distance migratory fish, return to specific spawning grounds for reproduction. To clarify reproductive neuroendocrine system of the tiger puffer, the changes in the expression levels of the genes encoding three gonadotropin-releasing hormones (GnRHs), gonadotropin-inhibitory hormone (GnIH), GnIH receptor (GnIH-R), kisspeptin and kisspeptin receptor in the brain and gonadotropin (GTH) subunits, growth hormone (GH) and prolactin (PRL) in the pituitary were examined in the tiger puffer captured in the wild at different reproductive stages, namely immature and mature fish of both sexes, and post-ovulatory females that were obtained by hormonal treatment. The amounts of three gnrh mRNAs, gnih, gnih-r, fshb and lhb were substantially increased in the mature fish compared to the immature fish, especially in the females, and these augmented expressions were drastically decreased in the post-ovulatory females. gh expression showed a slight increase in the mature males. In contrast, kiss2, kiss2r and prl did not show significant changes in the males but significantly decreased in the post-ovulatory females. The present results demonstrate the expression dynamics of the hypothalamo-pituitary-gonadal axis genes associated with the reproductive conditions and the possible involvement of the GnRH/GnIH/GTH system in the regulation of the sexual maturation and spawning in the wild tiger puffer.

Puffer Fish Gut Microbiota Studies Revealed Unique Bacterial Co-Occurrence Patterns and New Insights on Tetrodotoxin Producers.

Tetrodotoxin (TTX) is a potent neurotoxin isolated mainly from toxic puffer fish. To date, the TTX biosynthetic mechanism inside its hosts remains unresolved. Here, we hypothesize the TTX synthesis relies on the host gut microbiota, including the neglected non-culturable bacteria. In these studies, we collected the gut contents from 5 puffer fish species of the genus Takifugu including one suspected hybrid species for gut microbiota study by 16S rRNA amplicon metagenomics approach. Their gut samples were divided into toxic and non-toxic groups based on the TTX concentrations in the livers detected by LC-MS/MS. Bacterial diversity studies showed that gut microbiota structures were significantly different between toxic and non-toxic species. Vibrio and Cyanobacteria centered at the gut bacterial co-occurrence network, suggesting their importance in TTX biosynthesis. The results of PICRUSt2 metagenomic prediction and gene set enrichment analysis provided new support of arginine-precursor required in TTX biosynthesis. This is the first study to profile the gut microbiota in toxic and non-toxic puffer fish species by 16S rRNA amplicon metagenomic approach, defining significant microbial co-occurrence patterns in their gut environment. Our data supported the proposed biosynthesis of TTX inside the hosts by their gut bacterial symbionts using arginine as a precursor.

Protein biomarkers associated with frozen Japanese puffer fish (Takifugu rubripes) quality traits.

This study was designed to investigate proteome changes in Japanese puffer fish (Takifugu rubripes) during short- and long-term frozen storage. In total, 1484 proteins were quantified, and 164 proteins were identified as differential abundance proteins (DAPs) in Japanese puffer fish from two frozen storage treatment groups (14 days and 60 days) compared with the fresh control group. Correlation analysis between the DAPs and quality traits of the puffer fish muscle showed that 106 proteins were correlated closely with colour and texture (hardness, elasticity, and chewiness). Bioinformatics analysis revealed and Western blot analysis verified that Putative prothymosin alpha species, Bridging integrator 3, NADH: the ubiquinone oxidoreductase subunit and Mx species are candidate biomarkers for puffer fish properties. This study offers valuable evidence to improve the quality control and monitoring of Japanese puffer fish during transportation and storage.

The paraneuronal gill neuroendocrine system in ocellated puffer fish Leiodon cutcutia.

Pseudobranchial neurosecretory system (PSNS) is the third Neuroendocrine (NE) system found in the gill region of fishes in close association with pseudobranch/carotid labyrinth/carotid gland and can suitably be placed under the category of "Diffused NE system (DNES)." The cells belonging to this system fall under the category of "Paraneurons," a concept proposed by Fujita and coworkers. It is found uniformly in all the catfish species and some other noncatfish group of teleosts as Atheriniformes, Channiformes, Perciformes, and Clupeiformes. The fishes, in which the PSNS is present, belong to different breathing habits. Most of these have the capacity to tolerate low O2 conditions. Leiodon cutcutia although not an air-breathing fish, is known to retain air in its stomach for varied periods when threatened. In an attempt to verify the veracity of this system in a fish of another peculiar breathing habit, ocellated puffer fish L. cutcutia (order Tetradontiformes) was investigated. The histological observations undertaken on L. cutcutia revealed the presence of a well-developed extrabranchial NE system. The findings are discussed in the light of the association of PSNS with chemosensory system and its evolution in fishes, especially in the view of the transition from aquatic to terrestrial life.

Tetrodotoxin and saxitoxin in two native species of puffer fish, Sphoeroides marmoratus and Lagocephalus lagocephalus, from NE Atlantic Ocean (Madeira Island, Portugal).

The presence in EU waters of invasive tetrodotoxin (TTX) -harbouring puffer fishes has been receiving increasingly attention due to potential new threats posed by this potent neurotoxin. The present study investigates the occurrence of tetrodotoxin, saxitoxin (STX), and their analogues in two native puffer fish species from the NE Atlantic. High TTX content was detected by LC-MS/MS in several tissues of the Guinean puffer Sphoeroides marmoratus from Madeira Island (Portugal), reaching concentrations as high as 15 mg TTX kg-1 in the digestive tract of a male specimen and 7.4 mg TTX kg-1 in gonads of a female specimen. Several TTX analogues were also detected, including the 4-epi-TTX, 4,9-Anhydro-TTX, 5- 11- deoxyTTX and 6,11-dideoxyTTX. Although at low levels, STX was detected in liver of the Oceanic puffer Lagocephalus lagocephalus. Trace levels of decarbamoylsaxitoxin (dcSTX) were also observed in L. lagocephalus. This study reports the presence of TTX and STX in native fish from EU waters, highlighting the need for a proper understating of the origin, distribution and fate of these toxins in NE Atlantic.

Comparison of Physicochemical Characteristics and Fibril Formation Ability of Collagens Extracted from the Skin of Farmed River Puffer (Takifugu obscurus) and Tiger Puffer (Takifugu rubripes).

Acid-soluble collagen (ASC) and pepsin-soluble collagen (PSC) from the skin of river puffer (ASC-RP and PSC-RP) and tiger puffer (ASC-TP and PSC-TP) were extracted and physicochemically examined. Denaturation temperature (Td) for all the collagens was found to be 25.5-29.5 °C, which was lower than that of calf skin collagen (35.9 °C). Electrophoretic patterns indicated all four samples were type I collagen with molecular form of (α1)2α2. FTIR spectra confirmed the extracted collagens had a triple-helical structure, and that the degree of hydrogen bonding in ASC was higher than PSC. All the extracted collagens could aggregate into fibrils with D-periodicity. The fibril formation rate of ASC-RP and PSC-RP was slightly higher than ASC-TP and PSC-TP. Turbidity analysis revealed an increase in fibril formation rate when adding a low concentration of NaCl (less than 300 mM). The fibril formation ability was suppressed with further increasing of NaCl concentration, as illustrated by a reduction in the turbidity and formation degree. SEM analysis confirmed the well-formed interwoven structure of collagen fibrils after 24 h of incubation. Summarizing the experimental results suggested that the extracted collagens from the skin of river puffer and tiger puffer could be considered a viable substitute to mammalian-derived collagens for further use in biomaterial applications.

Effects of high and low temperature on expression of GnIH, GnIH receptor, GH and PRL genes in the male grass puffer during breeding season.

Gonadotropin-inhibitory hormone (GnIH) is a multifunctional hypophysiotropic neurohormone and has a stimulatory role in the control of reproduction in the grass puffer. To clarify the neuroendocrine mechanisms underlying the effect of changes in water temperature on reproduction in fish, we previously revealed that, in parallel to gonadal regression, both low and high temperature significantly decreased the expressions of the genes encoding kisspeptin (kiss2), kisspeptin receptor (kiss2r), gonadotropin-releasing hormone 1 (gnrh1) in the brain and gonadotropin (GTH) subunits (fshb and lhb) in the pituitary of sexually mature male grass puffer. In this study, we examined the changes in expression of gnih and GnIH receptor gene (gnihr) in the brain and pituitary along with the genes for growth hormone (gh) and prolactin (prl) in the pituitary of male grass puffer exposed to low temperature (14 °C), normal temperature (21 °C, as initial control) and high temperature (28 °C) conditions for 7 days. The levels of gnih and gnihr mRNAs were significantly decreased in both low and high temperature conditions compared to normal temperature in the brain and pituitary. Similarly, the gh mRNA levels were significantly decreased in both low and high temperature conditions. The prl mRNAs showed no significant changes at high temperature, whereas drastically decreased at low temperature possibly by dysfunctional cold stress. Taken together, the present results suggest that, in addition to the inhibitory effect of temperature changes on the Kiss2/GnRH1/GTH system, the suppression of GnIH/GH system may also be involved in the termination of reproduction by high temperature at the end of breeding season.