Amazons Are Back: Absence of Males in a Praying Mantis from Uruguayan Savannas.

With a broad distribution throughout South America, Brunneria subaptera (Saussure) (Coptopterygidae) is the most abundant praying mantis species in Uruguay, mostly associated with grassland vegetation. Their body coloration can vary between green or brown sepia. The recent catalog of Uruguayan mantids showed that all specimens of B. subaptera from the collections in Uruguay were females, suggesting an absence of males in natural populations. The aim of this study was to determine the occurrence of female-only populations of B. subaptera in Uruguay. Moreover, we analyzed the genetic variation and estimated the time of the most recent common ancestor of the species. We performed bimonthly samplings in two localities in Uruguay for 1 year and we sexed the individuals. We extracted DNA from the abdominal tissue of adult females and sequenced a fragment of the mitochondrial cox1 gene to assess the genetic diversity and to estimate diversification times. We did not find any males in either of the studied localities. The age estimates revealed a recent origin of the species (2.33 Ma), and there was no genetic differentiation between the individuals from Uruguayan localities. This absence of males of B. subaptera documented in the sampled locations suggests that the species reproduces by thelytokous parthenogenesis. This study opens promising venues for future research into reproductive strategies and polymorphism in South-American praying mantises that inhabit one of the largest areas of grasslands in the world, currently under dramatic deterioration and reduction.

Ritualized ink use during visual courtship display by males of the sexually dimorphic cuttlefish Sepia andreana.

Visual display is a crucial aspect of courtship, and their success relies on both display quality and the surrounding environment, such as the visual background. Cephalopods may release ink when attacked by predators or during aggressive interactions with conspecifics. Here, we report that ink is used as a part of the courtship display by males of the cuttlefish species Sepia andreana. Males of this species engage in a highly ritualized multimodal courtship using a pair of markedly long sexually dimorphic arms. At the climax of the courtship, the male releases a diffuse backdrop of ink near himself and then performs the specific courtship display by extending his sexually dimorphic arms and altering his body pattern to pale in front of this ink backdrop, and then proceeds to mate. This novel use of cephalopod ink could make the surroundings darker and more homogeneous, potentially serving as a temporary modification of the visual environment for courtship display.

Separable Microneedle for Integrated Hyperglycemia Sensing and Photothermal Responsive Metformin Release.

Microdevices that offer hyperglycemia monitoring and controllable drug delivery are urgently needed for daily diabetes management. Herein, a theranostic separable double-layer microneedle (DLMN) patch consisting of a swellable GelMA supporting base layer for glycemia sensing and a phase-change material (PCM) arrowhead layer for hyperglycemia regulation has been fabricated. The Cu-TCPP(Fe)/glucose oxidase composite and 3,3',5,5'-tetramethylbenzidine coembedded in the supporting base layer permit a visible color shift at the base surface in the presence of glucose via a cascade reaction, allowing for the in situ detection of glucose in interstitial fluid. The PCM arrowhead layer is encapsulated with water monodispersity melanin nanoparticles from Sepia officinalis and metformin that is imparted with a near-infrared ray photothermal response feature, which is beneficial to the controllable release of metformin for suppression of hyperglycemia. By applying the DLMN patch to the streptozotocin-induced type 2 diabetic Sprague-Dawley rat model, the results demonstrated that it can effectively extract dermal interstitial fluid, read out glucose levels, and regulate hyperglycemia. This DLMN-integrated portable colorimetric sensor and self-regulated glucose level hold great promise for daily diabetes management.

Comparative transcriptomic analysis of the different developmental stages of ovary in the cuttlefish Sepia pharaonis.

The cuttlefish, Sepia pharaonis, is characterized by rapid growth and strong disease resistance, making it an important commercially farmed cephalopod species in the southeastern coastal regions of China. However, in the reproductive process of S. pharaonis, there are challenges such as a low output of eggs, poor quality, and low survival rates of newly hatched juveniles. Therefore, there is an urgent need to study the molecular mechanisms underlying ovarian development in this species. In this study, we conducted the first transcriptomic analysis of the ovary at four developmental stages: the undeveloped stage, developing stage, nearly-ripe stage, and ripe stage, and compared the transcriptomics among these four stages using Illumina sequencing technology. The total numbers of clean reads of the four stages ranged from 40,890,772 to 52,055,714 reads. A total of 136,829 DEGs were obtained, GC base ratios of raw data were between 38.44 and 44.59%, and the number of uniquely mapped reads spanned from 88.08 to 95.90%. The Pearson correlation coefficient demonstrated a strong correlation among different samples within the same group, PCA and Anosim analysis also revealed that the grouping of these four stages was feasible, and each stage could be distinguished from the others. GO enrichment analysis demonstrated that ovarian follicle growth, sex differentiation, and transforming growth factor beta receptor, played a foreshadowing role at the early ovarian development stage, and the terms of small molecule metabolic process, peptide metabolic process, and catalytic activity were prominent at the mature stage. Meanwhile, KEGG analysis showed that the early ovarian development of S. pharaonis was mainly associated with the cell cycle, DNA replication, and carbon metabolism, while the mid-late ovarian development was involved with the signal transduction, endocrine system, and reproduction pathway. RT-qPCR further confirmed the consistent expression patterns of genes such as 17ß-HSD, GH, VGS, NFR, and NYR in the ovaries of S. pharaonis, exhibiting elevated levels of expression during the maturation stage. Conversely, ER and OM exhibited high expression levels during the early stages of ovarian development. These transcriptomic data provide insights into the molecular mechanisms of S. pharaonis ovarian development. The findings of this study will contribute to improving the reproduction and development of cuttlefish and enriching the bioinformatics knowledge of cephalopods.

Comparison of Cd, Cu, Se, and Zn Concentration in the Muscle and Hepatopancreas of Sepia pharaonis and Uroteuthis duvauceli in the North of Persian Gulf (Iran).

The objective of this research was to examine and contrast the levels of cadmium (Cd), copper (Cu), zinc (Zn), and selenium (Se) in the muscle and hepatopancreas tissues of two species, namely pharaoh cuttlefish (Sepia pharaonis) and Indian squid (Uroteuthis duvauceli), from the Persian Gulf. A total of thirty individuals of each species were gathered in January 2009 from the northern waters of the Persian Gulf. The metal concentrations were significantly higher in muscle tissue (p < 0.05) than in other tissues. S. pharaonis had higher metal concentrations than U. duvauceli. In the muscle and hepatopancreas samples of S. pharaonis, the highest mean concentrations were found to be for Zn (58.45 ± 0.96 µg/g dw) and Cu (1541.47 ± 192.15 µg/g dw), respectively. In U. duvauceli, the highest concentration of measured elements was seen for Zn in both muscle (36.52 ± 0.56 µg/g dw) and hepatopancreas (60.94 ± 2.65 µg/g dw). Se had the lowest concentration among the elements measured in both species. There was a negative and significant correlation between Cu and biometrical factors (total body length and weight) in both muscle and hepatopancreas samples of S. pharaonic and only in the muscle samples of U. duvauceli (p < 0.01, R2 = - 052; p < 0.01, R2 = - 0.055). However, there was a strong correlation between Zn and biometrical factors in hepatopancreas samples of both species. The comparison of metal concentrations with standards revealed that only Cd levels in S. pharaonis exceeded the ESFA and WHO standards, whereas other metals were below the standards.

Comparative proteomic analysis of the protein profile in the cuttlefish Sepia pharaonis associated with skin ulceration syndrome.

Skin ulceration syndrome (SUS) is becoming a severe problem in the breeding and culturing process of the cuttlefish Sepia pharaonis. However, limited knowledge is available about the occurrence of this devastating disease. In this study, proteomic analysis was used to identify the differentially expressed proteins (DEPs) and the biological pathways enriched in SUS-diseased S. pharaonis. Both the healthy group and diseased group were analyzed in triplicate, with 4 cuttlefish in each replicate. The results showed that 85 DEPs were identified between the two groups, including 36 upregulated proteins and 49 downregulated proteins in the diseased group compared to the healthy group. GO enrichment analysis revealed that the DEPs were mainly enriched in cellular component organization or biogenesis, nucleus and ion binding processes. The results of the KEGG pathway analysis indicated that extracellular matrix (ECM)-receptor interaction was the most enriched upregulated pathway. Real-time reverse transcriptase PCR was used to identify the expression of two differentially expressed matrix metalloproteinases (MMPs), and the results showed that the mRNA expression of MMP14 and MMP19 was significantly upregulated in the skin tissue of the diseased group. Furthermore, the protease activity of the diseased group was higher than that of the healthy group. Our results offer basic knowledge on the changes in protein profiles during the occurrence of SUS in the cuttlefish S. pharaonis.

Expansion of the genetic toolbox for manipulation of the global crop pest Drosophila suzukii: Isolation and assessment of eye colour mutant strains.

Drosophila suzukii (Matsumura) (Diptera: Drosophilidae), commonly called spotted wing Drosophila, is an important agricultural pest recognised worldwide. D. suzukii is a pest of soft-skinned fruits as females can lay eggs in ripening fruit before harvest. While strains for genetic biocontrol of D. suzukii have been made, the development of transgenic D. suzukii strains and their further screening remain a challenge partly due to the lack of phenotypically trackable genetic-markers, such as those widely used with the model genetic organism D. melanogaster. Here, we have used CRISPR/Cas9 to introduce heritable mutations in the eye colour genes white, cinnabar and sepia, which are located on the X, second and third chromosomes, respectively. Strains were obtained, which were homozygous for a single mutation. Genotyping of the established strains showed insertion and/or deletions (indels) at the targeted sites. A strain homozygous for mutations in cinnabar and sepia showed a pale-yellow eye colour at eclosion but darkened to a sepia colour after a week. The fecundity and fertility of some of the cinnabar and sepia strains were comparable with the wild type. Although white mutant males were previously reported to be sterile, we found that sterility is not fully penetrant and we have been able to maintain white-eyed strains for over a year. The cinnabar, sepia and white mutant strains developed in this study should facilitate future genetic studies in D. suzukii and the development of strains for genetic control of this pest.

Homeopathic Medicines in the Management of Dermatophytosis (Tinea Infections): A Clinico-epidemiological Study with Pre-post Comparison Design.

BACKGROUND: Dermatophytosis is a common fungal infection of the skin and nails. Insufficient data exist regarding the clinico-epidemiological profile of dermatophytosis and the usefulness of individualized homeopathic medicines (IHMs) for patients visiting a homeopathy outpatient department (OPD). OBJECTIVES: This article undertakes a clinico-epidemiological profiling of dermatophytosis and the usefulness of IHMs in its management. METHODS: This open-label, pre-post, comparative observational study was conducted in a homeopathy OPD from November 2018 to February 2020. IHMs were prescribed based on symptom totality and repertorization. A numeric rating scale (NRS) and the Dermatology Life Quality Index (DLQI) patient questionnaires were used, and results were analyzed using SPSS-IBM version 20. RESULTS: Data from a total of 103 patients, mean age 29.65 ± 15.40 years, were analyzed. Tinea cruris was the most common infection (29.1%), followed by tinea corporis (13.6%). After 3 months of treatment, significant reductions in NRS and DLQI scores were observed (8.51 ± 1.24 to 0.59 ± 0.83, p < 0.001, and 16.28 ± 5.30 to 1.44 ± 1.56, p < 0.001, respectively), with Sepia (15.5%), Sulphur (14.6%), Calcarea carbonica (11.7%), Natrum muriaticum (9.7%) and Bacillinum (8.7%) being the most frequently prescribed medicines. There was no significant correlation between occupation, sex, home location or marital status and the clinical types of dermatophytosis. No adverse events were reported. CONCLUSION: T. cruris and T. corporis were prevalent dermatophytic infections. The decrease in NRS and DLQI scores associated with homeopathy indicates its usefulness as an integrative treatment option for dermatophytosis. Further research in larger and more diverse population samples is needed.

Antioxidant Activity of an Aqueous Extract of Cuttlefish Ink during Fish Muscle Heating.

The antioxidant effect of cuttlefish (Sepia officinalis) ink (CFI) was analysed in the present study. A model system consisting of minced seabream (Sparus aurata) muscle and different concentrations of an aqueous extract of CFI was subjected to a heat (50 °C) treatment for 12 days. The effects of the CFI content and the heating time on lipid oxidation (conjugated diene (CD), conjugated triene (CT), and peroxide values and fluorescent compound formation), hydrolysis (free fatty acid content) development, and changes in the fatty acid (FA) profile (polyene index (PI), unsaturated FA content, ω3/ω6 ratio) were determined. The addition of the aqueous extract of CFI led to a lower (p < 0.05) development of lipid oxidation (CD, CT, and fluorescent compound determination) and to a higher (p < 0.05) retention of unsaturated FAs (PI determination). More important effects were found with increased CFI concentrations and at advanced heating times. However, a definite effect on lipid hydrolysis development (FFA value) could not be inferred. A new approach for the beneficial use of cuttlefish ink is presented. According to the direct relationship between rancidity stability and nutritional and sensory values, the present study provides a new strategy for the quality enhancement of thermally treated seafood.

Comparative Trophic Levels of Phragmocone-Bearing Cephalopods (Nautiloids, Ammonoids, and Sepiids).

Cephalopods are among many marine animals that through some combination of habit and/or habitat have proven difficult to study, especially understanding their trophic positions in marine communities. Stable isotope analyses have provided powerful tools for discovering quantitative aspects about the ecology and food sources of many cephalopod species. Here, we present new gut content and isotopic data (carbon and nitrogen isotopes) from phragmocone-bearing cephalopods (both ectocochleates, as well as those with internal, hard part buoyancy maintenance apparatuses). To this, we also include observations from baited remote underwater video systems to describe feeding habits and potential prey types to correlate with gut contents analyses. These data come from extant Allonautilus, Nautilus, and Sepia species, as well as from extinct nautiloids and ammonites. Extant nautiloids occupy a different isotopic niche than all other cephalopod groups where such data have been published to date. We conclude that these species are obligate scavengers rather than predators on any living species in their environments. Extant Nautilus and Allonautilus also demonstrate different patterns of functional allometry of nitrogen isotope values over ontogeny than do most other cephalopods (or animals in general), by showing decreasing nitrogen isotope levels during ontogeny. This pattern is shown to be different in Sepia and the yet small number of ammonite cephalopods studied to date, supporting the increasingly accepted view that ammonites were far closer to coleoids in basic biology than nautiloids. Overall, phragmocone-bearing cephalopods appear fundamentally different ecologically than cephalopods without this kind of buoyancy system. Of these groups, nautiloids appear to live a low-energy existence that allows them to subsist on energy-poor food sources, such as crustacean molts, as well as being able to scavenge in low oxygen basins where rare food falls, such as dead fish, remain unobtainable by most other animals.

Gene network analyses of Sepia esculenta larvae exposed to copper and cadmium: A comprehensive investigation of oxidative stress, immune response, and toxicological mechanisms.

Copper (Cu) and Cadmium (Cd), prevalent heavy metals in marine environments, have known implications in oxidative stress, immune response, and toxicity in marine organisms. Sepia esculenta, a cephalopod of significant economic value along China's eastern coastline, experiences alterations in growth, mobility, and reproduction when subjected to these heavy metals. However, the specific mechanisms resulting from heavy metal exposure in S. esculenta remain largely uncharted. In this study, we utilized transcriptome and four oxidative, immunity, and toxicity indicators to assess the toxicological mechanism in S. esculenta larvae exposed to Cu and Cd. The measurements of Superoxide Dismutase (SOD), Malondialdehyde (MDA), Glutathione S-Transferase (GST), and Metallothioneins (MTs) revealed that Cu and Cd trigger substantial oxidative stress, immune response, and metal toxicity. Further, we performed an analysis on the transcriptome data through Weighted Gene Co-expression Network Analysis (WGCNA) and Protein-Protein Interaction (PPI) network analysis. Our findings indicate that exposure methods and duration influence the type and the extent of toxicity and oxidative stress within the S. esculenta larvae. We took an innovative approach in this research by integrating WGCNA and PPI network analysis with four significant physiological indicators to closely examine the toxicity and oxidative stress profiles of S. esculenta upon exposure to Cu and Cd. This investigation is vital in decoding the toxicological, immunological, and oxidative stress mechanisms within S. esculenta when subjected to heavy metals. It provides foundational insights capable of advancing invertebrate environmental toxicology and informs S. esculenta artificial breeding practices.

Investigating the Mechanism of Low-Salinity Environmental Adaptation in Sepia esculenta Larvae through Transcriptome Profiling.

Sepia esculenta is an economically important mollusk distributed in the coastal waters of China. Juveniles are more susceptible to stimulation by the external environment than mature individuals. The ocean salinity fluctuates due to environmental changes. However, there is a lack of research on the salinity adaptations of S. esculenta. Therefore, in this study, we investigated the differential expression of genes in S. esculenta larvae after stimulation by low salinity. RNA samples were sequenced and 1039 differentially expressed genes (DEGs) were identified. Then, enrichment analysis was performed using the Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases. Finally, a protein-protein interaction network (PPI) was constructed, and the functions of key genes in S. esculenta larvae after low-salinity stimulation were explored. We suggest that low salinity leads to an excess proliferation of cells in S. esculenta larvae that, in turn, affects normal physiological activities. The results of this study can aid in the artificial incubation of S. esculenta and reduce the mortality of larvae.

The hydrodynamics of jet propulsion swimming in hatchling and juvenile European common cuttlefish, Sepia officinalis.

Cuttlefish swim using jet propulsion, taking a small volume of fluid into the mantle cavity before it is expelled through the siphon to generate thrust. Jet propulsion swimming has been shown to be more metabolically expensive than undulatory swimming, which has been suggested to be due to the lower efficiency of jet propulsion. The whole-cycle propulsive efficiency of cephalopod molluscs ranges from 38 to 76%, indicating that in some instances jet propulsion can be relatively efficient. Here, we determined the hydrodynamics of hatchling and juvenile cuttlefish during jet propulsion swimming to understand the characteristics of their jets, and whether their whole-cycle propulsive efficiency changes during development. Cuttlefish were found to utilise two jet types: isolated jet vortices (termed jet mode I) and elongated jets (leading edge vortex ring followed by a trailing jet; termed jet mode II). The use of these jet modes differed between the age classes, with newly hatched animals nearly exclusively utilising mode I jets, while juveniles showed no strong preferences. Whole-cycle propulsive efficiency was found to be high, ranging from 72 to 80%, and did not differ between age classes. During development, Strouhal number decreased as Reynolds number increased, which is consistent with animals adjusting their jetting behaviour in order to maximise whole-cycle propulsive efficiency and locomotor performance. Although jet propulsion swimming can have a relatively high energetic cost, in cuttlefish and nautilus, both neutrally buoyant species, the whole-cycle propulsive efficiency is actually relatively high.

A nanoscale study of the structure and electrical response of Sepia eumelanin.

Eumelanin, the brown-black member of the melanin biopigment family, is a prototype material for sustainable (green) organic electronics. Sepia eumelanin (Sepia) is a type of biosourced eumelanin extracted from the ink sac of cuttlefish. Electron microscopy and scanning probe microscopy images of Sepia show distinguishable near spherical granules with diameters of about 150-200 nm. We have recently reported on predominant electronic transport in printed films of Sepia formulated inks including the (insulating) binder Polyvinyl-butyral (PVB). In that work, we proposed that inter-granular percolative transport, observed for micrometric interelectrode distances, is promoted by the confining action of the PVB binder on the Sepia granules. Considering that inter-granular transport implies intra-granular transport, in this work we proceeded to a nanoscale study of Sepia granules by High Resolution Atomic Force Microscopy (HR-AFM) and Conductive-AFM (c-AFM). We have observed protrusions on the surface of the Sepia granules, suggesting sub-granular structures compatible with the hierarchical development of Sepia, as proposed elsewhere. For films of Sepia formulated inks deposited on gold-coated substrates, c-AFM revealed, for the very first time, a nanoscale electrical response. Nanoscale studies provide the key to structure-property relationships in biosourced materials strategic for sustainable organic electronics.

Preliminary study on toxicological mechanism of golden cuttlefish (Sepia esculenta) larvae exposed to cd.

BACKGROUND: Cadmium (Cd) flows into the ocean with industrial and agricultural pollution and significantly affects the growth and development of economic cephalopods such as Sepia esculenta, Amphioctopus fangsiao, and Loligo japonica. As of now, the reasons why Cd affects the growth and development of S. esculenta are not yet clear. RESULTS: In this study, transcriptome and four oxidation and toxicity indicators are used to analyze the toxicological mechanism of Cd-exposed S. esculenta larvae. Indicator results indicate that Cd induces oxidative stress and metal toxicity. Functional enrichment analysis results suggest that larval ion transport, cell adhesion, and some digestion and absorption processes are inhibited, and the cell function is damaged. Comprehensive analysis of protein-protein interaction network and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis was used to explore S. esculenta larval toxicological mechanisms, and we find that among the 20 identified key genes, 14 genes are associated with neurotoxicity. Most of them are down-regulated and enriched to the neuroactive ligand-receptor interaction signaling pathway, suggesting that larval nervous system might be destroyed, and the growth, development, and movement process are significantly affected after Cd exposure. CONCLUSIONS: S. esculenta larvae suffered severe oxidative damage after Cd exposure, which may inhibit digestion and absorption functions, and disrupt the stability of the nervous system. Our results lay a function for understanding larval toxicological mechanisms exposed to heavy metals, promoting the development of invertebrate environmental toxicology, and providing theoretical support for S. esculenta artificial culture.

Transcriptomic analysis of oxidative stress mechanisms induced by acute nanoplastic exposure in Sepia esculenta larvae.

Nanoplastics (NPs), as a new type of pollutant with a size small than 1 µm, are ubiquitous and harmful to organisms. There has been an increasing amount of research concerning the effects of NPs on organisms over recent years, especially on aquatic animals. However, there is a limited study on the impact of NPs on mollusk cephalopods. In this research, Sepia esculenta, belonging to Cephalopoda, Coleoidea, Sepioidea, was selected to explore the effects caused by NPs exposure. The S. esculenta larvae were exposed to polystyrene NPs (PS-NPs) with diameter 50 nm (100 mg/L) for 4 h. The detection of oxidative stress biomarkers displayed an obvious increase in SOD (superoxide dismutase) activity and MDA (malondialdehyde) level. Then, RNA-Seq was performed to explore the oxidative stress response at mRNA level. The transcriptome analysis demonstrated that the expression of 2,570 genes was affected by PS-NPs. Besides, the signaling pathways of ribosome, ribosome biogenesis in eukaryotes, proteasome, and MAPK were enriched. This study not only provides novel references for understanding the mechanisms of oxidative stress response induced by NPs, but also reminds us to follow with interest the influence of acute exposure to NPs.

Variation of 210-polonium in the cephalopod community from the Bay of Biscay, North-East Atlantic.

Among natural radionuclides, 210Po is the major contributor to the radiation dose received by marine organisms. In cephalopods, 210Po is concentrated in the digestive gland, which contains over 90% of the whole-body burden of the nuclide. Although previous studies showed that 210Po was taken up independently of 210Pb, its parent nuclide, very little is known about the factors influencing its levels in cephalopods. To the best of our knowledge, no studies investigated 210Po levels in different species at the same time. In the present study, 210Po was analysed in the digestive gland of 62 individuals from 11 species representing a large range of feeding ecologies and habitats, including squids, cuttlefish and octopus species from coastal to deep-oceanic habitats. Among species, the highest activity was measured in Loligo vulgaris (5720 ± 3606 Bq/kg) and the lowest in T. megalops (188 Bq/kg). However, considering the habitats (benthic vs pelagic and neritic vs oceanic), no significant differences appeared. At the species level, no differences between sexes were found so both sexes were plotted together to test the size effect for species with at least 8 individuals (i.e., Eledone cirrhosa, L. vulgaris, L. forbesi and Sepia officinalis). In the first three species, 210Po levels decreased significantly with increasing size or weight but not in S. officinalis. In squid, this could be related to ontogenetic changes in diet from a high proportion of crustaceans (high Po content) in small individuals to fish (low Po content) in larger individuals, while the high dietary plasticity of S. officinalis at all stages of its life cycle could explain the lack of decrease in 210Po with size. In comparison to the few data from the literature, the levels of 210Po concentrations in the cephalopod community of the Bay of Biscay were overall in the same range than those reported in other cephalopods, varying across 4 orders of magnitude. Further studies are needed to understand the mechanism of retention in the cephalopod digestive gland.

Understanding Why Homeopathic Medicines are Used for Menopause: Searching for Insights into Neuroendocrine Features.

BACKGROUND: Menopause is a physiological event that marks the end of a woman's reproductive stage in life. Vasomotor symptoms and changes in mood are among its most important effects. Homeopathy has been used for many years in treating menopausal complaints, though clinical and pre-clinical research in this field is limited. Homeopathy often bases its prescription on neuropsychiatric symptoms, but it is unknown if homeopathic medicines (HMs) exert a neuroendocrine effect that causes an improvement in vasomotor symptoms and mood during menopause. OBJECTIVES: The study's objectives were to address the pathophysiological changes of menopause that could help in the understanding of the possible effect of HMs at a neuroendocrine level, to review the current evidence for two of the most frequently prescribed HMs for menopause (Lachesis mutus and Sepia officinalis), and to discuss the future directions of research in this field. METHODS: An extensive literature search for the pathophysiologic events of menopause and depression, as well as for the current evidence for HMs in menopause and depression, was performed. RESULTS: Neuroendocrine changes are involved in the pathophysiology of vasomotor symptoms and changes in mood during menopause. Gonadal hormones modulate neurotransmitter systems. Both play a role in mood disorders and temperature regulation. It has been demonstrated that Gelsemium sempervirens, Ignatia amara and Chamomilla matricaria exert anxiolytic effects in rodent models. Lachesis mutus and Sepia officinalis are frequently prescribed for important neuropsychiatric and vasomotor symptoms. Dopamine, a neurotransmitter involved in mood, is among the constituents of the ink of the common cuttlefish, Sepia officinalis. CONCLUSION: Based on all the pathophysiologic events of menopause and the improvement in menopausal complaints that certain HMs show in daily practice, these medicines might have a direct or indirect neuroendocrine effect in the body, possibly triggered via an as-yet unidentified biological mechanism. Many unanswered questions in this field require further pre-clinical and clinical research.

Accidental discovery of a Tetraodontidae (Sphoeroides marmoratus) within a cuttlefish (Sepia officinalis) bought in a fish shop in Italy: risk assessment associated with the presence of Tetrodotoxin.

The discovery of a pufferfish specimen (Tetraodontidae) inside a frozen cuttlefish, purchased by a fishmonger, and caught in the Eastern Central Atlantic (FAO 34) is reported. The consumer, who reported this case to FishLab (Department of Veterinary Sciences, University of Pisa) for investigation, was a student of Veterinary Medicine at the University of Pisa. He recognized the Tetraodontidae because he attended practical lessons on fish morphological identification during the course of food inspection and was aware of the risks to human health linked to the Tetrodotoxin (TTX). In this study, the pufferfish was identified morphologically, using the FAO morphological keys, and molecularly, analyzing two markers, the cytochrome oxidase I (COI) and the cytochrome b genes, by DNA barcoding. The pufferfish was identified morphologically as Sphoeroides spp., and molecularly as Sphoeroides marmoratus using the COI gene (99-100% identity values). Literature reports that S. marmoratus from the Eastern Atlantic contains high concentrations of TTX in the gonads and the digestive tract. However, the possible passage of TTX from fish to other organisms linked to contact or ingestion has never been reported. This represents the first case of a potentially toxic pufferfish entering the market inside another organism. The fact that a student observed this occurrence highlights the key role of citizen science in the management of emerging risks.

Microplastics in Sepia officinalis caught on the central Adriatic coast: preliminary results.

Microplastics (µPs) represent an emerging problem for the marine environment given their wide bioavailability for all aquatic organisms, from zooplankton to top predators. This work aimed to evaluate a method of extracting microplastics from the gastrointestinal tract of 122 Sepia officinalis caught from the Adriatic Sea (along Abruzzo region coasts) to measure its quantity in a poorly investigated species. The extraction method used for gastrointestinal content was performed using 10% potassium hydroxide. In 98/122 (80.32-95% CI=73.27-87.37%) wild animals microplastics were detected with a mean concentration of 6.82±5.52 µPs/subject. Among the fragments, as supported by various authors, those of black color were the most represented ones; however, also blue fibers and transparent spheres were isolated. This study, in agreement with previous ones, poses further attention to the wide microplastic diffusion in the marine environment (surface, columns, sediments, and animals). The obtained results provide the basis for future investigations on this public health concern.