Synthesis of a novel magnetic calcium-rich biochar nanocomposite for efficient removal of phosphate from aqueous solution.

Phosphorus (P) scarcity and eutrophication have triggered the development of new materials for P recovery. In this work, a novel magnetic calcium-rich biochar nanocomposite (MCRB) was prepared through co-precipitation of crab shell derived biochar, Fe2+ and Fe3+. Characteristics of the material demonstrated that the MCRB was rich in calcite and that the Fe3O4 NPs with a diameter range of 18-22 nanometers were uniformly adhered on the biochar surface by strong ether linking (C-O-Fe). Batch tests demonstrated that the removal of P was pH dependent with an optimal pH of 3-7. The MCRB exhibited a superior P removal performance, with a maximum removal capacity of 105.6 mg g-1, which was even higher than the majority lanthanum containing compounds. Study of the removal mechanisms revealed that the P removal by MCRB involved the formation of hydroxyapatite (HAP-Ca5(PO4)3OH), electrostatic attraction and ligand exchange. The recyclability test demonstrated that a certain level (approximately 60%) was still maintained even after the six adsorption-desorption process, suggesting that MCRB is a promising material for P removal from wastewater.

Ghost crab (Ocypode quadrata) modify their behavior during mirror exposure.

Recently, ghost crabs (Ocypode quadrata) were shown to exhibit behavior consistent with passing the mark test, which implies the capacity for self-recognition. Considering this, it seems reasonable to suggest these animals can also distinguish their mirror image from different social stimuli, such as live conspecifics. In the current research, ghost crabs were placed in an experimental tank bisected by a mirror, clear partition separating a conspecific, and non-reflective partition on separate trials. Aggressive threat displays, time spent near partitions, eye cleaning, and partition contact were recorded during each condition. Results showed more aggression in trials with a conspecific present (i.e. clear partition), though this effect was only marginally significant. In addition, eye cleaning was significantly lower in the clear, compared to mirror and non-reflective treatments. Significantly more contact occurred with non-reflective partitions and time near each barrier was similar. These findings might suggest the capacity to distinguish mirror reflections from conspecifics, and mirror self-recognition could promote such ability. This, and other alternatives, are discussed as potential explanations for the observed effects.

Identification of ETH receptor and its possible roles in the mud crab Scylla paramamosain.

Ecdysis-triggering hormone (ETH) is a neuropeptide hormone characterized by a conserved KxxKxxPRx amide structure widely identified in arthropods. While its involvement in the regulation of molting and reproduction in insects is well-established, its role in crustaceans has been overlooked. This study aimed to de-orphanise a receptor for ETH in the mud crab Scylla paramamosain and explore its potential impact on ovarian development. A 513-amino-acid G protein-coupled receptor for ETH (SpETHR) was identified in S. paramamosain, exhibiting a dose-dependent activation by SpETH with an EC50 value of 75.18 nM. Tissue distribution analysis revealed SpETH was in the cerebral ganglion and thoracic ganglion, while SpETHR was specifically expressed in the ovary, hepatopancreas, and Y-organ of female crabs. In vitro experiments demonstrated that synthetic SpETH (at a concentration of 10-8 M) significantly increased the expression of SpVgR in the ovary and induced ecdysone biosynthesis in the Y-organ. In vivo experiments showed a significant upregulation of SpEcR in the ovary and Disembodied and Shadow in the Y-organ after 12 h of SpETH injection. Furthermore, a 16-day administration of SpETH significantly increased 20E titers in hemolymph, gonadosomatic index (GSI) and oocyte size of S. paramamosain. In conclusion, our findings suggest that SpETH may play stimulatory roles in ovarian development and ecdysone biosynthesis by the Y-organ.

Infestation of Lithodes santolla by Eremitione tuberculata: spatial and temporal variations in parasite prevalence and effect on host growth.

The southern king crab (SKC) Lithodes santolla is a crustacean parasitised by the bopyrid Eremitione tuberculata. This study aimed to analyse spatial and temporal variations in E. tuberculata prevalence in the juvenile SKC population of San Jorge Gulf (SJG) and adjacent waters (Argentine Patagonia), and evaluate the effects of the parasite on SKC juveniles to improve our understanding of its impact as a disease on SKC health condition. Moult increment and body weight were compared between parasitised and unparasitised individuals. The prevalence of E. tuberculata in SKC juveniles varied both spatially and temporally. In the south of SJG, the prevalence was 54.5% (n = 11). Temporal prevalence analysis revealed values lower than 17.4% in mid SJG during May and September 2015. No significant differences were observed in E. tuberculata prevalence between sexes or among seasons. Eremitione tuberculata had a negative effect on SKC growth (lower body dry mass, moult increment and relative increment rate) in parasitised individuals. We hypothesised that the higher prevalence of E. tuberculata in the south SJG could be attributed to the retention of parasite larvae and the presence of the frontal system in this part of the gulf. The temporal variations could reflect host mortality. Our results suggest that bopyrid infestation may have a more important role than previously believed in the dynamics of the SKC population in mid-Patagonia.

Membrane-targeting, ultrashort lipopeptide acts as an antibiotic adjuvant and sensitizes MDR gram-negative pathogens toward narrow-spectrum antibiotics.

Globally, infections due to multi-drug resistant (MDR) Gram-negative bacterial (GNB) pathogens are on the rise, negatively impacting morbidity and mortality, necessitating urgent treatment alternatives. Herein, we report a detailed bio-evaluation of an ultrashort, cationic lipopeptide 'SVAP9I' that demonstrated potent antibiotic activity and acted as an adjuvant to potentiate existing antibiotic classes towards GNBs. Newly synthesized lipopeptides were screened against ESKAPE pathogens and cytotoxicity assays were performed to evaluate the selectivity index (SI). SVAP9I exhibited broad-spectrum antibacterial activity against critical MDR-GNB pathogens including members of Enterobacteriaceae (MIC 4-8 mg/L), with a favorable CC50 value of ≥100 mg/L and no detectable resistance even after 50th serial passage. It demonstrated fast concentration-dependent bactericidal action as determined via time-kill analysis and also retained full potency against polymyxin B-resistant E. coli, indicating distinct mode of action. SVAP9I targeted E. coli's outer and inner membranes by binding to LPS and phospholipids such as cardiolipin and phosphatidylglycerol. Membrane damage resulted in ROS generation, depleted intracellular ATP concentration and a concomitant increase in extracellular ATP. Checkerboard assays showed SVAP9I's synergism with narrow-spectrum antibiotics like vancomycin, fusidic acid and rifampicin, potentiating their efficacy against MDR-GNB pathogens, including carbapenem-resistant Acinetobacter baumannii (CRAB), a WHO critical priority pathogen. In a murine neutropenic thigh infection model, SVAP9I and rifampicin synergized to express excellent antibacterial efficacy against MDR-CRAB outcompeting polymyxin B. Taken together, SVAP9I's distinct membrane-targeting broad-spectrum action, lack of resistance and strong in vitro andin vivopotency in synergism with narrow spectrum antibiotics like rifampicin suggests its potential as a novel antibiotic adjuvant for the treatment of serious MDR-GNB infections.

Differences in environmental microbial community responses under rice-crab co-culture and crab monoculture models under cyanobacterial bloom.

Cyanobacterial blooms (CBs) present significant challenges to Chinese mitten crab (CMC) culture, posing hazards to the aquatic microbial ecology. However, the current focus on the microbial ecological changes within the CMC culture system under the influence of CBs is somewhat insufficient. There's an urgent need to analyze the microbial ecosystem of the CMC culture system under CBs. This study employed 16S rRNA gene amplicon sequencing to investigate the dynamics of the environmental microbial community in both the rice-crab co-culture (RC) and crab monoculture (CM) models. The results revealed that cyanobacteria reached high levels in the CM water in July, while they began to increase in the RC water in August. Notably, OTU147 (uncultured bacterium g_Planktothrix NIVA-CYA 15), identified as the dominant taxon associated with CBs, showed a significant linear relationship with TP, NO2 --N, and the N:P ratio. TP, TN, NO2 --N, and CODMn had a more pronounced impact on the structure of bacterial communities and cyanobacterial taxa in the water. The bacterial community structure involved in carbon metabolism displayed temporal succession in the water. The co-occurrence network of the bacterial community primarily consisted of Chloroflexi, Proteobacteria, and Firnicutes in the sediment, and Actinobacteria, Proteobacteria, Chloroflexi, and Bacteroidota in the water. In contrast, the co-occurrence network included different peripheral species in the sediment and water. Keystone species were predominantly represented by OTU22 (uncultured actinobacterium g_ hgcI clade) and OTU12 (uncultured Opitutae bacterium g_ norank) in the RC water, and by OTU25 (uncultured bacterium g_ Limnohabitans) in the CM water. TP, TN, NO2 --N, and CODMn were identified as the primary environmental factors influencing these keystone taxa within the culture water. In conclusion, this study on the microbial ecology of the CMC culture system under the influence of CBs provides valuable insights that can be instrumental in subsequent management efforts.

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.

Ih Block Reveals Separation of Timescales in Pyloric Rhythm Response to Temperature Changes in Cancer borealis.

Motor systems operate over a range of frequencies and relative timing (phase). We studied the contribution of the hyperpolarization-activated inward current (Ih) to frequency and phase in the pyloric rhythm of the stomatogastric ganglion (STG) of the crab, Cancer borealis as temperature was altered from 11°C to 21°C. Under control conditions, the frequency of the rhythm increased monotonically with temperature, while the phases of the pyloric dilator (PD), lateral pyloric (LP), and pyloric (PY) neurons remained constant. When we blocked Ih with cesium (Cs+) PD offset, LP onset, and LP offset were all phase advanced in Cs+ at 11°C, and the latter two further advanced as temperature increased. In Cs+ the steady state increase in pyloric frequency with temperature diminished and the Q10 of the pyloric frequency dropped from ~1.75 to ~1.35. Unexpectedly in Cs+, the frequency displayed non-monotonic dynamics during temperature transitions; the frequency initially dropped as temperature increased, then rose once temperature stabilized, creating a characteristic "jag". Interestingly, these jags were still present during temperature transitions in Cs+ when the pacemaker was isolated by picrotoxin, although the temperature-induced change in frequency recovered to control levels. Overall, these data suggest that Ih plays an important role in the ability of this circuit to produce smooth transitory responses and persistent frequency increases by different mechanisms during temperature fluctuations.

The potential range of west Asian apple species Malus orientalis Uglitzk. under climate change.

The wild relatives of cultivated apples would be an ideal source of diversity for breeding new varieties, which could potentially grow in diverse habitats shaped by climate change. However, there is still a lack of knowledge about the potential distribution of these species. The aim of the presented work was the understand the impacts of climate change on the potential distribution and habitat fragmentation of Caucasian crab apple (Malus orientalis Uglitzk.) and the designation of areas of high interest according to climatic conditions. We used the MaxEnt models and Morphological-Spatial Analysis (MSPA) to evaluate the potential distribution, suitability changes, habitat fragmentation, and connectivity throughout the species range in Turkey, Armenia, Georgia, Russia, and Iran. The results revealed that the potentially suitable range of M. orientalis encompasses 858,877 km², 635,279 km² and 456,795 km² under the present, RCP4.5 and RCP8.5 scenario, respectively. The range fragmentation analysis demonstrated a notable shift in the edge/core ratio, which increased from 50.95% in the current scenario to even 67.70% in the future. The northern part of the range (Armenia, northern Georgia, southern Russia), as well as the central and western parts of Hyrcania will be a core of the species range with suitable habitats and a high connectivity between M. orientalis populations and could work as major refugia for the studied species. However, in the Zagros and central Turkey, the potential range will shrink due to the lack of suitable climatic conditions, and the edge/core ratio will grow. In the southern part of the range, a decline of M. orientalis habitats is expected due to changing climatic conditions. The future outlook suggests that the Hyrcanian forest and the Caucasus region could serve as important refuges for M. orientalis. This study helps to understand spatial changes in species' range in response to climate change and can help develop conservation strategies. This is all the more important given the species' potential use in future breeding programs aimed at enriching the gene pool of cultivated apple varieties.

Phytochemical Composition and Antimicrobial Efficacy of Salvadora persica Root Extracts Against Carbapenem-Resistant Acinetobacter baumannii.

Background Carbapenem-resistant Acinetobacter baumannii (CRAB) are difficult to eradicate from the environment and are virtually immune to all antibiotics. Consequently, CRAB may culminate in severe outbreaks and fatal infections among people attending hospitals and nursing homes. Salvadora persica has been used as an herbal remedy and chewing sticks for dental cleansing. Evaluating S. persica's efficacy against CRAB may provide an alternative approach to treating CRAB infections in healthcare environments, considering its traditional application in dental hygiene. Employing S. persica as an herbal remedy could be a part of a more sustainable approach to control CRAB infections. Aim To investigate the phytochemical composition of S. persica and evaluate its antimicrobial properties. Materials and methods The roots were extracted by Soxhlet apparatus using n-hexane, chloroform, and methanol. Each extract was analyzed using gas chromatography-mass spectrometry (GCMS) and characterized using WN908.L and National Institute of Standards and Technology (NIST) libraries. The antimicrobial activity of each extract against CRAB was evaluated using a broth microdilution assay to determine the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). Results The GCMS analysis of different solvent extracts of S. persica roots showed the presence of various phytochemical compounds such as steroids, phenolic compounds, fatty acids, alcohols, terpenoids, and vitamin E. Both chloroform and hexane extracts showed the most effective antimicrobial activity with a MIC value of 3.13 mg/mL and an MBC value of 12.50 mg/mL, respectively. Benzoic acid was the major phytochemical compound identified from S. persica extract. N-hexane, chloroform, and methanol extracts exhibited maximum antimicrobial activity due to the presence of active compounds in them. Conclusion Chloroform and hexane extracts showed the most potent antibacterial activities against CRAB.

Microplastics and 17α Ethinylestradiol: How Do Different Aquatic Invertebrates Respond to This Combination of Contaminants?

The synthetic hormone 17α ethinyl estradiol (EE2) is a molecule widely used in female contraceptives and recognized as a contaminant of attention (Watch List) in the European Union due to its high consumption, endocrine effects and occurrence in aquatic environments. Its main source of introduction is domestic sewage where it can be associated with other contaminants such as microplastics (MPs). Due to their characteristics, they can combine with each other and exacerbate their isolated effects on biota. This study evaluated the combined effects of microplastics (MPs) and 17α ethinylestradiol (EE2) on two tropical estuarine invertebrate species: Crassostrea gasar and Ucides cordatus. Polyethylene particles were spiked with EE2 and organisms were exposed to three treatments, categorized into three groups: control group (C), virgin microplastics (MPs), and spiked microplastics with EE2 (MPEs). All treatments were evaluated after 3 and 7 days of exposure. Oysters exhibited changes in phase 2 enzymes and the antioxidant system, oxidative stress in the gills, and reduced lysosomal membrane stability after exposure to MPs and MPEs. Crabs exposed to MPs and MPEs after seven days showed changes in phase 1 enzymes in the gills and changes in phases 1 and 2 enzymes in the hepatopancreas, such as disturbed cellular health. The combined effects of microplastics and EE2 increased the toxicity experienced by organisms, which may trigger effects at higher levels of biological organization, leading to ecological disturbances in tropical coastal ecosystems.

Allergenicity and Linear Epitope Analysis of Scy p 8, an Allergen from Mud Crab.

Scy p 8 (triosephosphate isomerase) as a crab allergen in inducing distinct T-helper (Th) cell differentiation and a linear epitope associated with allergenicity remain elusive. In this study, mice sensitized with Scy p 8 exhibited significantly upregulated levels of IgE, IgG1, and IL-4 release, inducing a Th2 immune response. Moreover, the release of IFN-γ (Th1) and the levels of Treg cells were downregulated, while IL-17A (Th17) was upregulated, indicating that Scy p 8 disrupted the Th1/Th2 balance and Th17/Treg balance in mice. Furthermore, bioinformatics prediction and serum samples from crab-allergic patients and mice enabled the discovery of 8 linear epitopes of Scy p 8. Meanwhile, the analysis of peptide similarity and tertiary superposition revealed that 8 epitopes of Scy p 8 exhibited conservation across various species, potentially resulting in cross-reactivity. These findings possess the potential to enhance the comprehension of crab allergens, thereby establishing a foundation for investigating cross-reactivity.

Characterization and efficacy against carbapenem-resistant Acinetobacter baumannii of a novel Friunavirus phage from sewage.

Carbapenem-resistant Acinetobacter baumannii (CRAB) has become a new threat in recent years, owing to its rapidly increasing resistance to antibiotics and new effective therapies are needed to combat this pathogen. Phage therapy is considered to be the most promising alternative for treating CRAB infections. In this study, a novel phage, Ab_WF01, which can lyse clinical CRAB, was isolated and characterized from hospital sewage. The multiplicity of infection, morphology, one-step growth curve, stability, sensitivity, and lytic activity of the phage were also investigated. The genome of phage Ab_WF01 was 41, 317 bp in size with a GC content of 39.12% and encoded 51 open reading frames (ORFs). tRNA, virulence, and antibiotic resistance genes were not detected in the phage genome. Comparative genomic and phylogenetic analyses suggest that phage Ab_WF01 is a novel species of the genus Friunavirus, subfamily Beijerinckvirinae, and family Autographiviridae. The in vivo results showed that phage Ab_WF01 significantly increased the survival rate of CRAB-infected Galleria mellonella (from 0% to 70% at 48 h) and mice (from 0% to 60% for 7 days). Moreover, after day 3 post-infection, phage Ab_WF01 reduced inflammatory response, with strongly ameliorated histological damage and bacterial clearance in infected tissue organs (lungs, liver, and spleen) in mouse CRAB infection model. Taken together, these results show that phage Ab_WF01 holds great promise as a potential alternative agent with excellent stability for against CRAB infections.

Phylogenetic inferences based on distinct molecular markers reveals a novel Babesia (Babesia pantanalensis nov. sp.) and a Hepatozoon americanum-related genotype in crab-eating foxes (Cerdocyon thous).

Piroplasmids and Hepatozoon spp. Are apicomplexan protozoa that may cause disease in several canid species. The present study aimed to expand the knowledge on the diversity of piroplasmids and Hepatozoon in crab-eating foxes (Cerdocyon thous; n = 12) sampled in the Pantanal of Mato Grosso do Sul State, central-western Brazil. PCR assays based on the 18S rRNA were used as screening. Three (25%) and 11 (91.7%) were positive for piroplasmids and Hepatozoon spp., respectively. Co-infection was found in three C. thous. Phylogenetic analyses based on the near-complete 18S rRNA, cox-1 and hsp70 genes evidenced the occurrence of a novel of Babesia spp. (namely Babesia pantanalensis nov. sp.) closely related to Rangelia vitalii and Babesia sp. 'Coco'. This finding was supported by the genetic divergence analysis which showed (i) high divergence, ranging from 4.17 to 5.62% for 18 S rRNA, 6.16% for hps70 and 4.91-9.25% for cox-1 and (ii) the genotype network (which displayed sequences separated from the previously described Piroplasmida species by median vectors and several mutational events). Also, phylogenetic analysis based on the 18S rRNA gene of Hepatozoon spp. positioned the sequences obtained herein in a clade phylogenetically related to Hepatozoon sp. 'Curupira 2', Hepatozoon sp. detected in domestic and wild canids from Uruguay and Hepatozoon americanum. The present study described Babesia pantanalensis nov sp. and Hepatozoon closely related to H. americanum in crab-eating foxes from Brazil. Moreover, the coinfection by piroplasmids and Hepatozoon sp. for the first time in crab-eating foxes strongly suggesting that this wild canid species potentially acts as a bio-accumulate of hemoprotozoan in wild environment.

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.

[Investigation of the population diversity of intermediate host snails and crabs of Paragonimus along Jiulong River, Zhangjiang River and Dongxi River basins in southern Fujian Province].

OBJECTIVE: To investigate the population distribution of intermediate host snails and crabs of Paragonimus along the Jiulongjiang River, Zhangjiang River, and Dongxi River basins in Bopingling Mountain, southern Fujian Province, so as to provide baseline data for researches on parasitic disease prevention and control and enlargement of samples in the parasitic resource bank. METHODS: A total of 23 villages in 8 counties (districts) along the Jiulong River, Zhangjiang River, and Dongxi River basins in Zhangzhou City, Fujian Province were selected as survey sites during the period from November 2020 through March 2023, and snail and freshwater crabs were sampled from 1 to 3 streams and ditches neighboring residential areas in each village. Morphological identification of snails was performed according to the external morphological characteristics of collected snail shells, and the unidentified snail species sampled from the natural foci of paragonimiasis in Yunxiao County were subjected to se-quence analysis of the mitochondrial cytochrome oxidase 1 (CO1) gene. The crab species was identified by observing the morphological characteristics of the terminal segment of the first pleopod of male crabs, and Paragonimus cercariae and metacercariae were detected in collected snails. RESULTS: The shells of the unidentified snails sampled from the natural foci of paragonimiasis in Yunxiao County were approximately 50 mm in height and 18 mm in width, thick and solid, long tower cone-shaped, and had 8 to 10 whorls. CO1 gene sequence analysis identified the snail species as Sulcospira hainanensis. A total of 6 freshwater snail species belonging to 5 genera within 3 families, identified 23 survey sites, including Semisulcospira libertina, Paludomus zhangchouensis and S. hainanensis that belonged to the Family Pleurceridae, Tricula fujianensis and T. huaanensis that belonged to the subfamily Triculinae, Family Pomatiopsidae, and Melanoides tuberculata (Family Thiaridae), and 11 species of freshwater crabs belonging to 5 genera within 2 families were identified, including Sinopotamon genus of S. jianglense, S. pinheense, and S. zhangzhouense, Huananpotamon genus of H. planopodum and H. zhangzhouense, Nanhaipotamon genus of N. huaanense and N. longhaiense, and Minpotamon genus of M. nasicum and M. auritum that belonged to the Family Potamidae, and Somanniathelphusa genus of S. huaanensis and S. zhangpuensis (Family Parathelphusidae). In addition, the prevalence of P. westermani cercariae infections was 0.08% (2/2 317) in P. zhangchouensis from Danyan Village in Changtai District and 0.09% (1/1 039) in S. hainanensis from Jinkeng Village in Yunxiao County, and the prevalence of P. westermani metacercariae infections was 25.81% (8/31) in S. jianglense from Danyan Village in Changtai District, and 26.31% (5/19) in S. zhangzhouense from Jinkeng Village in Yunxiao County, respectively. CONCLUSIONS: There is a population diversity in the intermediate host snails and crabs along the Jiulongjiang River, Zhangjiang River, and Dongxi River basins in Bopingling Mountain, southern Fujian Province, and P. zhangzhouensis and S. hainanensis are, for the first time, confirmed as the first intermediate hosts of P. westermani.

Potential Food Inclination of Crab-Eating Macaques in Laboratory Environments: Enhancing Positive Reinforcement Training and Health Optimization.

Positive reinforcement and training for health optimization are pivotal for successful studies with monkeys. Potential food inclination is important for studies on crab-eating macaques in laboratory environments, but evaluations remain scarce. We explored crab-eating macaques' potential food inclination to establish a reward system for future behavioral assessments. Twelve male and three female monkeys underwent a food inclination assessment in which they were offered four food categories-fruits, vegetables, proteins, and nuts. The monkeys exhibited a higher inclination for plant-based foods, particularly fruits and vegetables, over animal-based proteins like chicken and tuna (p < 0.0001), with a notable inclination for nuts (eaten/provided = 100%). Additionally, the consistency of potential food inclination after repeated offerings was investigated, revealing a time-dependent increase in inclination for protein items. Food consumption ratios correlated positively with caloric intake (r = 0.59, p = 0.02), implying that individuals with a regular high caloric intake and increased body weight are more likely to accept food during positive reinforcement training. Our findings suggest fruits, vegetables, protein-rich foods, and nuts can help with health optimization. However, animal-based protein-rich foods initially had a low preference, which may increase over time. Our study can provide guidelines for positive reinforcement training and health optimization.

When Nature Requires a Resource to Be Used-The Case of Callinectes sapidus: Distribution, Aggregation Patterns, and Spatial Structure in Northwest Europe, the Mediterranean Sea, and Adjacent Waters.

The Atlantic blue crab Callinectes sapidus, which is native to the western Atlantic coast and listed among the 100 most invasive alien species in the Mediterranean Sea, is attracting a great deal of interest because of its rapid colonisation of new areas, the significant increase in its population, and the impacts it may have on ecosystems and ecosystem services. Outside its natural distribution range, the species was first found on European Atlantic coasts in the early 1900s and was introduced into the Mediterranean Sea a few decades later, probably through ballast water. Currently, it is found in almost the entire Mediterranean Basin and is also expanding into the Black Sea and along the north African and Iberian Atlantic coasts. Based on a systematic review of C. sapidus occurrences, this study describes its distribution, aggregation patterns, and spatial structure in Northwest Europe, the Mediterranean Sea, and adjacent waters through a series of ecological indicators elaborated using GIS spatial-temporal statistics. The main results highlight that the species is expanding in the Mediterranean and adjacent waters, while in northern Europe, the population remains confined in some areas. Furthermore, the main species detection methods are analysed, finding that traps and nets are the most frequently used methods, and management suggestions are provided.

Targeting TtVgR via siRNA Knockdown Elicits Ovarian Cell Death in the Tri-spine Horseshoe Crab.

The vitellogenin present in the bloodstream undergoes internalization into developing oocytes through the vitellogenin receptor (VgR), a process mediated by receptor-mediated endocytosis. VgR plays a crucial role in facilitating the accumulation of vitellogenin and the maturation of oocytes. In this study, we characterized a Tachypleus tridentatus vitellogenin receptor (TtVgR) gene from the tri-spine horseshoe crab, revealing a length of 1956 bp and encoding 652 amino acid residues with 12 exons. TtVgR has a molecular weight of 64.26 kDa and an isoelectric point of 5.95. Predictions indicate 85 phosphorylation sites and 7 glycosylation sites within TtVgR. Transcriptional analysis demonstrated specific expression of TtVgR in the ovary and yellow connective tissue. TtVgR was identified and distributed in the plasma membrane of oocytes. The siRNA-mediated TtVgR knockdown significantly reduced the transcriptional activity of TtVgR. This depletion induced excessive ROS production, resulting in DNA damage in ovarian primary cells. TUNEL and flow cytometry analyses confirmed ovarian cell apoptosis following TtVgR knockdown, indicating DNA damage in ovarian primary cells. These findings underscore the importance of TtVgR in ovarian cell development, suggesting its potential involvement in vitellogenesis and oocyte maturation. This knowledge may inform innovative breeding strategies and contribute to the sustainable management and conservation of the tri-spine horseshoe crab.