Search

Pufferfish is common in marine, brackish and freshwater habitats in Bangladesh. Pufferfish poisoning sporadically occurs due to the consumptions of toxic pufferfish throughout the country. A study was conducted for molecular characterization and phylogenetic analysis of Bangladeshi pufferfish using two markers, mitochondrial cytochrome c oxidase I (COI) and 16S ribosomal RNA (16S rRNA) genes. A total of 34 barcode sequences (19 COI & 15 16S) from 30 collected specimens representing six species and four genera. The mean length of the sequences were 644 bp and 613 bp for COI and 16S, respectively. The average %GC content was 50.06 for COI which was higher compared to the 16S rRNA (46.61). Average Kimura two parameter (K2P) distances within species was found as 0.29 ± 0.01 for COI and 0.70 ± 0.06 for 16S rRNA. The Maximum Likelihood (ML) phylogenetic analysis of COI and 16S rRNA gene sequences revealed that the corresponding species were clustered together in the same clade. Cluster sequence analyses result revealed five Operational Taxonomic Units (OTUs). DNA based identification of the rest puffer species of Bangladesh with multiple sequences and phylogenetic relationship remains to be established.

A previously unreported heterodetic cyclic peptide, homophymamide A (1), was isolated from a Homophymia sp. marine sponge. The structure of homophymamide A was determined to be a lower homologue of anabaenopeptins by spectroscopic analysis, chemical degradation, and chemical synthesis. Analysis of the acidic hydrolysate showed that the racemization of Lys took place, leading us to pose a cautionary note on the configurational assignment of peptides that contain a ureido bond.

Myrindole A, a bis-indole alkaloid, was isolated from the deep-sea sponge Myrmekioderma sp. The high degree of unsaturation of the molecule complicated the assignment of its structure by standard 2D-NMR experiments but was ultimately achieved by a combination of 1H-15N-HMBC and 1,n-ADEQUATE experiments as well as the comparison of measured and calculated CD spectra. Myrindole A showed antimicrobial activity against Gram-positive and Gram-negative bacteria.

Reliable information about degree of red coloration in fruit flesh is essential for grading and sorting of red-fleshed apples. We propose a spatially resolved interactance spectroscopy approach as a new rapid and non-destructive technique to estimate degree of red coloration in the flesh of a red-fleshed apple cultivar ‘Kurenainoyume’. A novel measurement system was developed to obtain spatially resolved interactance spectra (190–1070 nm) for apple fruits at eight different light source-detector separation (SDS) distances on fruit surface. Anthocyanins in apple were extracted using a solvent extraction technique, and their contents were quantified with a spectrophotometer. Partial least squares (PLS) regression analyses were performed to develop estimation models for anthocyanin content from spatially resolved interactance spectra. Results showed that the PLS models based on interactance spectra obtained at different SDS distances achieved different predictive accuracy. Further, the system demonstrated the possibility to detect the degree of red coloration in the flesh at specific depths by identifying an optimal SDS distance. This might contribute to provide a detailed profile of the red coloration (anthocyanins) that is unevenly distributed among different depths of the flesh. This new approach may be potentially applied to grading and sorting systems for red-fleshed apples in fruit industry.

Toxic crabs of the family Xanthidae contain saxitoxins (STXs) and/or tetrodotoxin (TTX), but the toxin ratio differs depending on their habitat. In the present study, to clarify within reef variations in the toxin profile of xanthid crabs, we collected specimens of the toxic xanthid crab Zosimus aeneus and their sampling location within a single reef (Yoshihara reef) on Ishigaki Island, Okinawa Prefecture, Japan, in 2018 and 2019. The STXs/TTX content within the appendages and viscera or stomach contents of each specimen was determined by instrumental analyses. Our findings revealed the existence of three zones in Yoshihara reef; one in which many individuals accumulate extremely high concentrations of STXs (northwestern part of the reef; NW zone), another in which individuals generally have small amounts of TTX but little STXs (central part of the reef; CTR zone), and a third in which individuals generally exhibit intermediate characteristics (southeastern part of the reef; SE zone). Furthermore, light microscopic observations of the stomach contents of crab specimens collected from the NW and CTR zones revealed that ascidian spicules of the genus Lissoclinum were dominant in the NW zone, whereas those of the genus Trididemnum were dominant in the CTR zone. Although the toxicity of these ascidians is unknown, Lissoclinum ascidians are considered good candidate source organisms of STXs harbored by toxic xanthid crabs., Marine Drugs, 19(12), art. no. 670; 2021

Background: Forecasting bud physiologic conditions can help ‘Fuji’ apple farmers manage their orchards more efficiently. Being able to determine the nature of a bud before bud burst is one such forecast that could be of use to these ‘Fuji’ growers. The aim of this research project was to determine if a device, a visible/near-infrared spectrometer, could be employed to determine whether a bud is a flower or non-flower bud without destroying the bud. Methods: Experiments were conducted on buds taken from a ‘Fuji’ apple tree, beginning on January 29 through to March 31, 2021, three days before bud burst. The data from the visible/near-infrared spectrometer clarified whether a bud was a flower or a non-flower bud. The Spectro data Classification Learner App proved to be an accurate classification method to analyze flower and non-flower bud Spectro data. Result: Three days before bud burst, the chlorophyll content levels of the non-flower buds were markedly higher (P≤0.05) than those of the flower bud, which explains why the visible border of the near-infrared spectrometer might have been changed by the chlorophyll content of buds. The visible and near-infrared bands of the buds showed that the Spectro data of the non-flower buds were higher than those of the flower buds when measurements were made three days before bud burst. Three days before bud burst Cubic KNN of KNN classifier analyzed flower and non-flower buds smoothly. Spectro data were labeled as accuracy 75.9%, sensitivity 86% and specificity 67%. The results that were obtained suggest that farmers could use a visible/near-infrared spectrometer to identify flower and non-flower buds in their orchards, without damaging the buds, three days before bud burst.

Being able to ascertain the physiological condition of the buds on a young apple tree before bud burst could help farmers manage their orchards more efficiently, especially if they could do so without destroying the buds in the process. The experiments carried out in this study were conducted with the aim of distinguishing shoot from non-shoot buds before bud burst using a visible/near-infrared spectrometer, a device that does not destroy the buds being tested. Tests on spring-planted (April 30, 2021) trees were conducted to check shoot and non-shoot bud physiology and the winter dormancy of young ‘Jonagold’, ‘Miyabi Fuji’ and ‘Orin’ apple trees. The light absorbance of the shoot buds before bud burst was much lower than the light absorbance of the non-shoot buds as checked on the visible/near-infrared spectrometer. The highest first factor effect was determined by a PCA test conducted on shoot and non-shoot ‘Jonagold’ buds (99.9%) at a range of 640-652 nm, ‘Miyabi Fuji’ buds (99.7%) at 654-680 nm and ‘Orin’ buds (99.6%) at 704-766 nm seven days before bud burst. We also found that the highest level of accuracy, using the Classifier analysis, between shoot and non-shoot ‘Jonagold’ buds (76.6%) was one day before bud burst, for ‘Miyabi Fuji’ buds (82.1%) it was three days before and for ‘Orin’ buds (76.3%) it was two days before. These findings suggest that growers can more effectively manage the development of the young trees in their orchards with a visible/near-infrared spectrometer.

Adults of the ectoparasitic copepod Caligus fugu found on tetrodotoxin (TTX)-bearing pufferfish such as Takifugu alboplumbeus and Takifugu flavipterus are known to accumulate TTX in body tissues and parts other than the ovaries, oviducts, eggs, and cuticles. This study aimed to demonstrate, using immunoenzymatic staining techniques, that the TTX-free planktonic/infective copepodid stage of C. fugu could accumulate TTX in the tissues after molting into the parasitic stage (chalimus I) and then fed on mucus of host puffers. All the tissues of the planktonic copepodids were completely TTX-free, whereas chalimus I copepods accumulated TTX in parts other than the cuticles, guts, and some muscles. Chalimus IV and adult copepods retained TTX in these body parts but not in the reproductive organs, which were TTX-resistant, indicating that TTX was not vertically transmitted via eggs. Non-cellular TTX-positive contents found in the guts of some chalimi and adults indicated that the copepods potentially accumulated TTX by feeding on host mucus rather than skin tissues and blood. This study revealed that the presence or absence of TTX in some body parts differed among individuals of the parasite.

We tested whether tetrodotoxin (TTX) functions as a stress relieving substance in puffer fish. We orally administered TTX to the juveniles of hatchery-reared non-toxic tiger puffer Takifugu rubripes and measured the effects of TTX on brain corticotropin-releasing hormone (CRH) mRNA expression and plasma cortisol levels in comparison with effects in non-toxic juveniles. Firstly, the reciprocal connections of CRH and adrenocorticotropic hormone (ACTH) were confirmed by dual-label immunohistochemistry. CRH-immunoreactive (ir) cell bodies were detected in the hypothalamus and CRH-ir fibers were observed to project to ACTH-ir cells in the rostral pars distalis of the pituitary. Next, a TTX-containing diet (2.35 mouse units (517?ng)/g diet) or a non-toxic diet were fed to the fish for 28 days under a recirculating system. Standard length and body weight became significantly larger in the TTX-treated group. The degree of loss of the caudal fin, which is an indicator of the degree of agonistic interactions, where high values show a higher loss of caudal fin of a fish due to nipping by other individuals, was significantly lower in the TTX-treated group. Relative CRH mRNA expression levels in the brain and cortisol levels in the plasma were significantly lower in the TTX-treated group. These results indicate that TTX functions as a stress relieving substance by affecting the CRH-ACTH-cortisol axis and reducing agonistic interactions in tiger puffer juveniles., Toxicon, 171, pp.54-61; 2019

This study estimates future changes in the early summer precipitation characteristics around Japan using changes in the large-scale environment, by combining Global Precipitation Measurement precipitation radar observations and phase 5 of the Coupled Models Intercomparison Project climate model large-scale projections. Analyzing satellite-based data, we first relate precipitation in three types of rain events (small, organized, and midlatitude), which are identified via their characteristics, to the large-scale environment. Two environmental fields are chosen to determine the large-scale conditions of the precipitation: the sea surface temperature and the midlevel large-scale vertical velocity. The former is related to the lower-tropospheric thermal instability, while the latter affects precipitation via moistening/drying of the midtroposphere. Consequently, favorable conditions differ between the three types in terms of these two environmental fields. Using these precipitation–environment relationships, we then reconstruct the precipitation distributions for each type with reference to the two environmental indices in climate models for the present and future climates. Future changes in the reconstructed precipitation are found to vary widely between the three types in association with the large-scale environment. In more than 90% of models, the region affected by organized-type precipitation will expand northward, leading to a substantial increase in this type of precipitation near Japan along the Sea of Japan, and in northern and eastern Japan on the Pacific side, where its present amount is relatively small. This result suggests an elevated risk of heavy rainfall in those regions because the maximum precipitation intensity is more intense in organized-type precipitation than in the other two types.

The red color of sweet cherries (Prunus avium L.) is important for their marketability. Anthocyanin synthesis in ‘Satohnishiki’ is highly dependent on light conditions, and ultraviolet (UV) light from 280 to 320 nm (UV-B) reportedly has a strong effect on anthocyanin synthesis in the cultivar ‘Satohnishiki’. The effect of visible light, such as blue light, on anthocyanin accumulation in sweet cherries has not yet been studied. This research examined the effect of blue light on exocarp color development and anthocyanin accumulation in ‘Satohnishiki’ and ‘Napoleon’ yellow flesh, blush sweet cherry cultivars. Using a light emitting diode, the effect of blue light on anthocyanin accumulation was compared with that of UV-B. The blue light of 450 nm induced anthocyanin accumulation and red color development in both cultivars, while its effect was lower than that of UV-B light. The efficiency of the blue light wavelength on anthocyanin accumulation in ‘Satohnishiki’ was examined with using 430 to 490 nm blue lights, and 450 nm was more effective than other blue lights. Blue light may play an important role in red color development in the fruits of both cultivars under a tree canopy.

A previously unreported heterodetic cyclic peptide, homophymamide A (

Myrindole A, a bis-indole alkaloid, was isolated from the deep-sea sponge

The species classification of Cambodian freshwater pufferfish is incomplete and confusing, and scientific information on their toxicity and toxin profile is limited. In the present study, to accumulate information on the phylogeny and toxin profile of freshwater pufferfish, and to contribute to food safety in Cambodia, we conducted simultaneous genetic-based phylogenetic and toxin analyses using freshwater pufferfish individuals collected from Phnom Penh and Kratie (designated PNH and KTI, respectively).Phylogenetic analysis of partial sequences of three mitochondrial genes (cytochrome b, 16S rRNA, and cytochrome c oxidase subunit I) determined for each fish revealed that PNH and KTI are different species in the genus Pao (designated Pao sp. A and Pao sp. B, respectively). A partial sequence of the nuclear tributyltin-binding protein type 2 (TBT-bp2) gene differentiated the species at the amino acid level. Instrumental analysis of the toxin profile revealed that both Pao sp. A and Pao sp. B possess saxitoxins (STXs), comprising STX as the main component. In Pao sp. A, the toxin concentration in each tissue was extremely high, far exceeding the regulatory limit for STXs set by the Codex Committee,whereas in Pao sp. B, only the skin contained high toxin concentrations. The difference in the STX accumulation ability between the two species with different TBT-bp2 sequences suggests that TBT-bp2 is involved in STX accumulation in freshwater pufferfish., Toxins, 12(11), art.no.689; 2020

Pufferfish of the family Tetraodontidae possess tetrodotoxin (TTX) and/or saxitoxins (STXs), but the toxin ratio differs, depending on the genus or species. In the present study, to clarify the distribution profile of TTX and STXs in Tetraodontidae, we investigated the composition and intra-body distribution of the toxins in Canthigaster valentini. C. valentini specimens (four male and six female) were collected from Amami-Oshima Island, Kagoshima Prefecture, Japan, and the toxins were extracted from the muscle, liver, intestine, gallbladder, gonads, and skin. Analysis of the extracts for TTX by liquid chromatography tandem mass spectrometry and of STXs by high-performance liquid chromatography with post-column fluorescence derivatization revealed TTX, as well as a large amount of STXs, with neoSTX as the main component and dicarbamoylSTX and STX itself as minor components, in the skin and ovary. The toxins were also detected in the other tissues, but in much lower amounts than in the skin and ovary. The TTX/STX ratio varied greatly, depending on the tissue, but TTX was the major toxin component in the whole body, and STXs accounted for 25% and 13% of the total toxin amount in males and females, respectively. Like the marine pufferfish of the genus Arothron, C. valentini should be considered a pufferfish with considerable amounts of both TTX and STXs present simultaneously., Toxins, 12(7), art.no.436; 2020

The inactivation and damage of histamine-forming bacteria (HFB), Enterobacter aerogenes and Staphylococcus capitis, in a 0.1 M potassium phosphate buffer (pH 6.8) and marlin meat slurry by high hydrostatic pressure (HHP) treatments were studied using viability measurement and scanning electron microscopy (SEM). HHP treatments showed first order destruction kinetics to E. aerogenes and S. capitis during the pressure holding period. HFB in marlin meat slurry had higher D values and were more resistant to HHP treatments than in phosphate buffer. In phosphate buffer, E. aerogenes had higher D values than S. capitis at &gt, 380 MPa of pressure, whereas the reverse trend was noticed at lower pressures (&lt, 380 MPa). In marlin meat slurry, S. capitis had a higher D value than E. aerogenes at the same treatment pressure, indicating that S. capitis was more resistant to HHP treatment. To our knowledge, this is the first report to demonstrate that HHP can be used to inactivate HFB, E. aerogenes, and S. capitis, by causing disruption to bacterial cell membrane and cell wall as demonstrated by SEM micrographs.

The intoxication mechanism of xanthid crabs and edible shore crabs in Japan by ‘paralytic shellfish toxin’ (PST) and/or ‘tetrodotoxin’ (TTX) was investigated. Three xanthid crabs were toxic and their toxin consisted of PST and/or TTX. Since great individual and regional variation in toxicity of each specimen, and toxin variety with exposure of toxic crab to each toxin were confirmed, their intoxication mechanism was suggested to be a food chain feeding toxic plankton. However, as they are omnivorous, the conclusion remains uncertain. Two edible shore crabs including Telmessus acutidents were contaminated with PST around 2003, in the Tohoku district of Japan. The intoxication mechanism of the crab was experimentally shown to be through the food chain by feeding on bivalves infested with PST-producing Alexandrium sp. Many food poisoning cases due to ingestion of xanthid crabs have occurred so far, but there are no reports in the case of this shore crab.

Stellatolide H (1) was isolated from a deep-sea sponge Discodermia sp. as the cytotoxic constituent. The planar structure of 1 was elucidated on the basis of the NMR spectroscopic and mass spectrometric data. The absolute configurations of the constituent amino acid residues were determined by the Marfey’s method. Stellatolide H (1) is a peptide lactone of the callipeltin class with its N-terminus blocked by 3-hydroxy-6,8-dimethyldeca-(4Z,6E)-dienoic acid (Hdda).

The present study evaluated differences in the tetrodotoxin (TTX)/saxitoxins (STXs) selectivity between marine and freshwater pufferfish by performing in vivo and in vitro experiments. In the in vivo experiment, artificially reared nontoxic euryhaline freshwater pufferfish Dichotomyctere fluviatilis were intrarectally administered a mixture of TTX (24 nmol/fish) and STX (20 nmol/fish). The amount of toxin in the intestine, liver, muscle, gonads, and skin was quantified at 24, 48, and 72 h. STX was detected in the intestine over a long period of time, with some (2.7–6.1% of the given dose) being absorbed into the body and temporarily located in the liver. Very little TTX was retained in the body. In the in vitro experiments, slices of intestine, liver, and skin tissue prepared from artificially reared nontoxic D. fluviatilis and the marine pufferfish Takifugu rubripes were incubated in buffer containing TTX and STXs (20 nmol/mL each) for up to 24 or 72 h, and the amount of toxin taken up in the tissue was quantified over time. In contrast to T. rubripes, the intestine, liver, and skin tissues of D. fluviatilis selectively took up only STXs. These findings indicate that the TTX/STXs selectivity differs between freshwater and marine pufferfish., Toxins, 13(10), art. no. 731; 2021

Tetrodotoxin (TTX), also known as pufferfish toxin, has been detected in marine edible bivalves worldwide. In this study, several bivalve species, Azumapecten farreri subsp. akazara, Patinopecten yessoensis and Mytilus galloprovincialis, collected from the Pacific side of the northern Japanese Islands, were studied for the accumulation of TTX in the presence of toxic planocerid larvae. LC-MS/MS analysis demonstrated that TTX was detected only in the midgut gland of A. farreri subsp. akazara. Toxic flatworm-specific PCR and direct sequencing of the amplicons showed that the DNA fragments of the Planocera multitentaculata COI gene were detected in the gut contents of the toxified bivalves. The planocerid larvae were also detected in the environmental seawaters. Toxification experiments in the aquarium demonstrated that the mussel M. galloprovincialis was also toxified by feeding on the toxic flatworm larvae. These results suggest that the source of TTX accumulation in edible bivalves is toxic flatworm larvae.