Acute toxicity, secondary metabolites, and antioxidant activity of Macaranga tanarius from post-coal mining and non-mining areas in East Kalimantan, Indonesia.

Coal plays a crucial role in Indonesia's foreign exchange and East Kalimantan's revenue sharing, yet its environmental impacts, including soil acidification, raises concerns. Reclamation measures involve revegetation with pioneer plants such as Macaranga sp., known for their medicinal properties. However, the pharmacological properties of these plants are influenced by secondary metabolites, which depend on soil parameters such as pH and nutrient levels. The aim of this study was to evaluate the acute toxicity, secondary metabolites, and antioxidant activities of Macaranga tanarius leaf extracts from post-coal mining area (MTPCMA) and non-mining area (MTNMA) alongside soil parameters. Acute toxicity of M. tanarius leaf extracts and soils were assessed using the brine shrimp lethality test (BSLT). Phytochemical screening was done using thin-layer chromatography (TLC), determining total phenolic (TPC) and flavonoid content (TFC). The DPPH radical scavenging assay was used to assess the antioxidant activity. A comparative analysis between MTPCMA and MTNMA was conducted using Student t-test. The data showed no significant difference in toxicity between MTPCMA and MTNMA leaf extracts (LC50 of 100-1000 µg/mL) (p=0.062), and soils from both areas were non-toxic (LC50 of >1000 µg/mL). Although heavy metal concentrations were higher in PCMA than in NMA soil (p<0.001), secondary metabolite compounds and TFC in both extracts were not significantly different (p=0.076). Both extracts contained flavonoids and polyphenols with antioxidant activity and terpenoids without antioxidant activities. The DPPH radical scavenging test suggested insignificant antioxidant activity between MTPCMA and MTNMA extracts (p=0.237). In conclusion, non-toxic soils in post-mining land and insignificant differences between MTPCMA and MTNMA extracts suggest good soil nutrient availability, highlighting the success of land recovery after 10 years of revegetation with M. tanarius.

Morphological, Toxicological, and Biochemical Characterization of Two Species of Gambierdiscus from Bahía de La Paz, Gulf of California.

We describe five new isolates of two Gambierdiscus species from Bahía de La Paz in the southern Gulf of California. Batch cultures of Gambierdiscus were established for morphological characterization using light microscopy (LM) and scanning electron microscopy (SEM). Pigment and amino acid profiles were also analyzed using high-performance liquid chromatography (HPLC-UV and HPLC-DAD). Finally, toxicity (CTX-like and MTX-like activity) was evaluated using the Artemia salina assay (ARTOX), mouse assay (MBA), marine fish assay (MFA), and fluorescent receptor binding assay (fRBA). These strains were identified as Gambierdiscus cf. caribaeus and Gambierdiscus cf. carpenteri. Toxicity for CTX-like and MTX-like activity was confirmed in all evaluated clones. Seven pigments were detected, with chlorophyll a, pyridine, Chl2, and diadinoxanthin being particularly noteworthy. For the first time, a screening of the amino acid profile of Gambierdiscus from the Pacific Ocean was conducted, which showed 14 amino acids for all strains except histidine, which was only present in G. cf. caribeaus. We report the presence of Gambierdiscus and Fukuyoa species in the Mexican Pacific, where ciguatera fish poisoning (CFP) cases have occurred.

Bioactive potential of Eugenia luschnathiana essential oil and extract: antifungal activity against Candida species isolated from oncological patients.

Immunosuppressed individuals, including those undergoing cancer treatment, are more vulnerable to fungal infections, such as oral candidiasis, impacting their quality of life. Given the limitations of current therapies, the discovery of new antifungal agents, including those of natural origin, is crucial for the proper managing of these infections. We investigated the phytochemical profile and antifungal activity of both the essential oil and crude ethanolic extract (CEE) obtained from Eugenia luschnathiana against reference strains and clinical isolates of Candida from oncology patients. Toxicological characterization was also conducted. Gas chromatography coupled to mass spectrometry (GC-MS) and 1H Nuclear Magnetic Resonance (NMR) were used for phytochemical analysis. Antifungal evaluation was conducted to determine the Minimum Inhibitory Concentration (MIC) and Minimum Fungicidal Concentration (MFC); evaluation of potential mechanisms of action; activity on a fungal biofilm; evaluation of the cytotoxic effect on human keratinocytes of the HaCat lineage by the MTT method; determination of lethality for Artemia salina larvae. GC-MS identified a predominance of sesquiterpenes in the essential oil, notably (E)-Caryophyllene. The 1H NMR spectrum identified aliphatic, osidic, and aromatic compounds in the crude ethanolic extract. The essential oil showed no antifungal activity. However, the CEE exhibited fungicidal activity, with MIC and MFC ranging from 1.95 µg/mL to 3.90 µg/mL. The antifungal effect was affected by sorbitol, indicating a possible mechanism targeting fungal cell wall structures. At low concentration (19.5 µg/mL), the CEE inhibited 62,78% of C. albicans biofilm. The CEE demonstrated a promising toxicity profile, with an LC50 of 142.4 µg/mL against Artemia salina. In conclusion, the CEE from Eugenia luschnathiana exhibited potent antifungal activity, likely through cell wall disruption, biofilm inhibition, and a favorable toxicity profile for further exploration.

Novel Fabrication of Bio-polymeric Nanogel Loaded with Nalidixic Acid Formulations for the Effective Prevention of Bacterial Pathogens Leading Urinary Tract Infection and Its Biosafety Evaluation.

Management of urinary tract infections (UTI) is a highly challenging process due to the biofilm-forming ability of human-pathogenic bacteria. Here, we designed to fabricate an effective nanogel with a combination of chitosan bio-polymer and nalidixic acid to prevent biofilm-forming bacterial pathogens. Chitosan-coated nalidixic acid nanogel (NA@CS) exhibits outstanding inhibition potential against bacterial strains. In vitro, anti-bacterial analysis methods (well diffusion, colony-forming assay, and anti-biofilm assay) were performed to study the bacterial inhibition potential of prepared nanogel, which reveals that NA@CS nanogel have greater inhibition potential against selected pathogens. The combination of nalidixic acid with chitosan biopolymer decreases the virulence and pathogenicity of biofilm-forming pathogens due to their ability to membrane phospholipids penetration. Furthermore, the fabricated NA@CS nanogel showed reliable in vitro bio-compatibility on L929 fibroblast cells and in vivo compatibility with Artemia salina animal model. Overall, the results demonstrate that NA@CS nanogel could be an effective therapeutic for treating urinary tract infections and urine bladder wound healing.

Optimization of Rhizoclonium hieroglyphicum extract for enhanced synthesis of nickel oxide nanoparticles using response surface methodology and its potential exploration in biological application.

The study investigates the potential of Rhizoclonium hieroglyphicum as a novel source for synthesizing nickel oxide nanoparticles (RH-NiONPs) and evaluates its biological applications. Phytochemicals in the algal extract serve as capping, reducing and stabilizing agent for nickel oxide nanoparticles. The process variables were optimized using BBD based RSM to obtain maximum RH-NiONPs. Characterization of RH-NiONPs using UV-Vis and FT-IR spectroscopy reveals the plasmon resonance peak at 340 nm and the functional groups responsible for reduction and stabilization. XRD confirmed the crystalline nature while the stability and size of the RH-NiONPs were determined by DLS and zeta potential. Toxicity assessments demonstrated the effect of RH-NiONPs against Vigna radiata, Allium cepa and Artemia salina was low. RH-NiONPs revealed significant zone of inhibition against the selected bacteria and fungi. The results of larvicidal activity showed that RH-NiONPs are toxic to 4th instar larvae of Daphnis nerii. Also, RH-NiONPs efficiently decolorized Reactive Violet 13 (92%) under sunlight irradiation and the experimental data well fits to Langmuir isotherm along with pseudo second order kinetic model. The thermodynamic studies enunciate the exothermic and non-spontaneous photocatalytic decolorization of reactive violet 13. Thus, the current study assesses the eco-friendly and cost-effective nature of RH-NiONPs along with its biological applications.

The coral pathogen Vibrio coralliilyticus uses a T6SS to secrete a group of novel anti-eukaryotic effectors that contribute to virulence.

Vibrio coralliilyticus is a pathogen of coral and shellfish, leading to devastating economic and ecological consequences worldwide. Although rising ocean temperatures correlate with increased V. coralliilyticus pathogenicity, the specific molecular mechanisms and determinants contributing to virulence remain poorly understood. Here, we systematically analyzed the type VI secretion system (T6SS), a contact-dependent toxin delivery apparatus, in V. coralliilyticus. We identified 2 omnipresent T6SSs that are activated at temperatures in which V. coralliilyticus becomes virulent; T6SS1 is an antibacterial system mediating interbacterial competition, whereas T6SS2 mediates anti-eukaryotic toxicity and contributes to mortality during infection of an aquatic model organism, Artemia salina. Using comparative proteomics, we identified the T6SS1 and T6SS2 toxin arsenals of 3 V. coralliilyticus strains with distinct disease etiologies. Remarkably, T6SS2 secretes at least 9 novel anti-eukaryotic toxins comprising core and accessory repertoires. We propose that T6SSs differently contribute to V. coralliilyticus's virulence: T6SS2 plays a direct role by targeting the host, while T6SS1 plays an indirect role by eliminating competitors.

Structural and functional alterations under stress conditions by contamination: A multi-species study in a non-forced multi-compartmented mesocosm.

Despite the existing connectivity and heterogeneity of aquatic habitats, the concept of interconnected landscapes has been frequently overlooked in ecotoxicological risk assessment studies. In this study, a novel mesocosm system, the HeMHAS (Heterogeneous Multi-Habitat Assay System), was constructed with the potential to assess structural and functional changes in a community resulting from exposure to contaminants, while also considering the complex ecological scenarios. Fish (Sparus aurata), shrimp (Palaemon varians) and three species of marine microalgae (Isochrysis galbana, Nannochloropsis gaditana and Tetraselmis chuii) were used as test organisms. Other species, such as Artemia sp. and macroalgae were also introduced into the system as environmental enrichment. All the species were distributed in five interconnected mesocosm compartments containing a copper gradient (0, 1, 10, 100 and 250 µg/L). The mobile fish avoided the copper contaminants from 1 µg/L (24 h-AC50: 4.88 µg/L), while the shrimp avoided from 50 µg/L (24 h-AC50: 136.58 µg/L). This finding suggests interspecies interactions influence habitat selection in contaminated environments, potentially jeopardizing population persistence. Among the non-motile organisms, the growth and chlorophyll content of the microalgae were concentration dependent. The growth of I. galbana was more sensitive (growth inhibition of 50 % at the highest concentration) in contrast to N. gaditana (30 % inhibition at the highest concentration) and T. chuii (25 % inhibition at the last two highest concentrations). In summary, the mesocosm HeMHAS showed how contamination-driven responses can be studied at landscape scales, enhancing the ecological relevance of ecotoxicological research.

Single-nucleus atlas of the Artemia female reproductive system suggests germline repression of the Z chromosome.

Our understanding of the molecular pathways that regulate oogenesis and define cellular identity in the Arthropod female reproductive system and the extent of their conservation is currently very limited. This is due to the focus on model systems, including Drosophila and Daphnia, which do not reflect the observed diversity of morphologies, reproductive modes, and sex chromosome systems. We use single-nucleus RNA and ATAC sequencing to produce a comprehensive single nucleus atlas of the adult Artemia franciscana female reproductive system. We map our data to the Fly Cell Atlas single-nucleus dataset of the Drosophila melanogaster ovary, shedding light on the conserved regulatory programs between the two distantly related Arthropod species. We identify the major cell types known to be present in the Artemia ovary, including germ cells, follicle cells, and ovarian muscle cells. Additionally, we use the germ cells to explore gene regulation and expression of the Z chromosome during meiosis, highlighting its unique regulatory dynamics and allowing us to explore the presence of meiotic sex chromosome silencing in this group.

In silico mechanistic insights of ecofriendly synthesized AgNPs, SeNPs, rGO and Ag&SeNPs@rGONM's for biological applications and its toxicity evaluation using Artemia salina.

The present study focused on Rosmarinus officinalis Linn. leaves extract (ROE) mediated synthesis of silver nanoparticles (AgNPs), selenium nanoparticles (SeNPs), reduced graphene oxide (rGO) and silver and selenium nanoparticles decorated on rGO nanomaterials (Ag&SeNPs@rGONM's) for its antibacterial and antifungal in silico mechanistic insight applications. In addition, the toxicity of the synthesized nanomaterials was evaluated using Artemia salina. The formation of AgNPs, SeNPs, rGO and Ag&SeNPs@rGONM's was completed within 1.0, 140, 120 and 144 h, respectively. Various optical and microscopic examinations were evident in the nanomaterial's synthesis. Further, the average size and stability of the synthesized nanomaterials were conformed through dynamic light scattering (DLS) and zeta potential analyzer, respectively. The synthesized Ag&SeNPs@rGONM's were pronounced promising results against Gram-negative bacteria of Escherichia coli and the results achieved from the route of entry and action, reactive oxygen species (ROS), and antioxidant nature of nanoparticles were evidence of its properties. Computational studies further supported these findings, indicating much of the phytochemicals present in ROE well interact with the bacterial surface proteins. Similarly, the synthesized Ag&SeNPs@rGONM's was effective against Fusarium graminearum and Alternaria alternata in a dose dependent manner than its original nanomaterials. In addition, the docking study also confirmed that rosmarinic acid and caffeic acid prominently interacted with the fungal proteins. Interestingly, Ag&SeNPs@rGONM's pronounced less toxic effect compared to AgNPs and SeNPs against Artemia salina, which shows its biocompatibility.

Source and variation of the amazing live Sea-Monkey microbiome.

An embryonic diapause in unfavourable conditions has allowed brine shrimp to thrive in hypersaline environments and, unexpectedly, mail-order sachets and small, novelty tanks. Marketed as Sea-Monkeys®, each kit involves a 3-step process to generate adult Artemia within a matter of weeks. Whether these kits also allow for the maintenance of a host-associated microbiome is unclear. Therefore, comparing five replicate tanks under the same culture conditions, we sequenced the 16S ribosomal small subunit (SSU) gene to analyse bacterial community compositions in adults, their surrounding tank water, and their feed. Adult Sea-Monkeys® harboured a bacterial microbiome that was clearly distinguishable from the tank water and food. Furthermore, individual tanks had a notable effect on fine-scale microbiome variation. Several Sea-Monkey bacterial variants appeared absent in environmental samples and included genera (Leucobacter and Microbacterium) known to confer desiccation resistance in other hosts. Although Sea-Monkeys® taxonomy is unclear, phylogenetic inference of the cytochrome c oxidase I (COXI) gene from the host animal suggests Sea-Monkeys® belong to the Artemia franciscana 'superspecies'. Overall, Sea-Monkeys® kits appear to be a convenient and scalable mesocosm for the study of host-microbiome interactions and could serve as a useful tool for future invertebrate microbiome research, outreach, and education.

Using decapsulated local brine shrimp cysts on feeding common carp larvae in hatcheries.

The current study aims at using non-hatchable artemia eggs of local origin and making use of these eggs by decapsulating and presenting them as food for the larvae of the Cyprinus carpio as a source of animal protein with high nutritional value instead of throwing them away. The results showed that the second parameter (A2) was highly significant at the level (P≤0.05) in the growth rates of the larvae that were fed on decapsulated artemia eggs alone, and it was better than the two control parameters (A1), in which the larvae were fed with feed designated for Cyprinus carpio fish. It also outperformed the third parameter (A3), in which the feed was mixed with artemia eggs with 50% decapsulation, which also outperformed the control parameter with high significance at the same level (P≤0.05).

In vitro-antibacterial properties of ten medicinal plants against common uropathogenic organisms and toxicity determination using brine shrimp lethality assay.

BACKGROUND: In recent years, antibiotic resistance has emerged as a global health concern in bacterial infections such as urinary tract infections (UTIs). Uropathogenic Escherichia coli is the most frequent organism responsible for both simple and complex UTIs. Staphylococcus aureus and Pseudomonas aeruginosa are frequently associated with complicated UTIs. Sri Lanka has significant resources of medicinal plants used to cure UTIs in Ayurvedic and traditional medicine. METHODS: Agar well diffusion and broth microdilution methods were used to determine the antibacterial activity of the methanolic extract of ten medicinal plants against P. aeruginosa ATCC27853, S.aureus ATCC25923, E.coli ATCC25922 and their UTI positive strains extracted from positive culture plates. As a preliminary toxicity assay, the Brine Shrimp Lethality Assay (BSLA) was used to determine its cytotoxicity. RESULTS: The methanolic fruits extract of P. emblica demonstrated the highest antibacterial activity against both E. coli ATCC25922 and E. coli UTI-positive strains. B. diffusa roots extract exhibited the highest activity against S. aureus ATCC25923, while T. chebula fruits extract showed the highest activity against the S. aureus UTI-positive strain. T. involucrata roots extract displayed the highest activity against P. aeruginosa ATCC27853, and Z. officinale rhizomes extract showed the highest activity against the P. aeruginosa UTI-positive strain. Moreover, the plant mixture showed the most substantial antibacterial effect against P. aeruginosa ATCC27853. However, the methanolic seed extract of C. melo did not exhibit any antimicrobial effects against the selected organisms. All plant material, including the plant mixture, showed cytotoxicity according to the BSLA. CONCLUSION: All the methanolic extracts including P. emblica fruits, O. tenuiflorum whole plant, T. chebula fruits, Z. officinale rhizome, T. terrestris roots, T. involucrata roots, A. lanata whole plant. B. diffusa roots and A. falcatus roots showed antimicrobial effects against selected strains except C. melo seed extract. The results of the present study evidently supports the traditional and ayurvedic use of these plants for the treatment of UTIs. This paves the way for another praise for new plant-based therapeutic product development for the treatment of UTIs. However, further toxicity studies are needed for medicinal dose determination.

Ecotoxicological studies of direct and indirect genotoxicity with Artemia: a integrative review.

Artemia is a brine shrimp genus adapted to extreme habitats like ranges salinity from 5-25 g/L and in temperatures from 9 to 35 °C. It is widely distributed and used as an environmental quality biomarker. Artemia franciscana and Artemia salina species are commonly used in ecotoxicological studies and genotoxicity assays due to their short life cycle, high fecundity rate, easy culture, and availability. Thus, considering the importance of these tests in ecotoxicological studies, the present study aimed to present Artemia genus as a biological model in genotoxicity research. To this end, we reviewed the literature, analyzing data published until July 2023 in the Web of Science, SCOPUS, Embase, and PubMed databases. After screening, we selected 34 studies in which the genotoxicity of Artemia for various substances. This review presents the variability of the experimental planning of assays and biomarkers in genotoxicity using Artemia genus as a biological model for ecotoxicological studies and show the possibility of monitoring biochemical alterations and genetic damage effects. Also highlight innovative technologies such as transcriptomic and metabolomic analysis, as well as studies over successive generations to identify changes in DNA and consequently in gene expression.

Antioxidant, antibacterial, and anti-biofilm activities of selected indigenous plant species against nosocomial bacterial pathogens.

Biofilms are responsible for over 60% of nosocomial infections. The focus of this study was to investigate the antioxidant, antibacterial, antibiofilm, and anti-motility activities of Gardenia volkensii, Carissa bispinosa, Peltophorum africanum, and Senna petersiana. Antioxidant activity was evaluated using free radical (DPPH) scavenging and ferric reducing power assays. Antibacterial and antibiofilm activities were evaluated using the broth micro-dilution and the crystal violet assays, respectively. Anti-motility was evaluated using anti-swarming activities, and the brine shrimp lethality assay was used for cytotoxicity. Gardenia volkensii and C. bispinosa acetone extracts had low EC50 values of 9.59 and 9.99 µg ml-1on the free-radical scavenging activity, respectively. All the plant extracts demonstrated broad-spectrum antibacterial activity against Klebsiella pneumoniae, Pseudomonasa aeruginosa, Escherichia coli, Enterococcus faecalis, and Staphylococcus aureus [minimum inhibitory concentration (MIC) < 0.63 mg ml-1]. The initial cell adherence stage of P. aeruginosa and E. coli was the most susceptible stage where sub-MICs resulted in inhibitions >50%. Peltophorum africanum had the least cytotoxic effects. All extracts had anti-motility activity against P. aeruginosa and E. coli. This study showed that not only do the plants have strong antibacterial activity but had noteworthy inhibition (>50%) of initial cell adherence and may be suitable candidates for the treatment of nosocomial pathogens.

Integrating ATAC-Seq and RNA-Seq Reveals the Signal Regulation Involved in the Artemia Embryonic Reactivation Process.

Embryonic diapause is a common evolutionary adaptation observed across a wide range of organisms. Artemia is one of the classic animal models for diapause research. The current studies of Artemia diapause mainly focus on the induction and maintenance of the embryonic diapause, with little research on the molecular regulatory mechanism of Artemia embryonic reactivation. The first 5 h after embryonic diapause breaking has been proved to be most important for embryonic reactivation in Artemia. In this work, two high-throughput sequencing methods, ATAC-seq and RNA-seq, were integrated to study the signal regulation process in embryonic reactivation of Artemia at 5 h after diapause breaking. Through the GO and KEGG enrichment analysis of the high-throughput datasets, it was showed that after 5 h of diapause breaking, the metabolism and regulation of Artemia cyst were quite active. Several signal transduction pathways were identified in the embryonic reactivation process, such as G-protein-coupled receptor (GPCR) signaling pathway, cell surface receptor signaling pathway, hormone-mediated signaling pathway, Wnt, Notch, mTOR signaling pathways, etc. It indicates that embryonic reactivation is a complex process regulated by multiple signaling pathways. With the further protein structure analysis and RT-qPCR verification, 11 GPCR genes were identified, in which 5 genes function in the embryonic reactivation stage and the other 6 genes contribute to the diapause stage. The results of this work reveal the signal transduction pathways and GPCRs involved in the embryonic reactivation process of Artemia cysts. These findings offer significant clues for in-depth research on the signal regulatory mechanisms of the embryonic reactivation process and valuable insights into the mechanism of animal embryonic diapause.

Clinacanthus nutans crude polysaccharide extract as a green platform for microwave-assisted synthesis of silver nanoparticles: Optimization, characterization, and evaluation of bioactivities.

Clinacanthus nutans (C. nutans) is a plant in tropical Asia with proven biological activities. The optimized extraction method of C. nutans crude polysaccharide (CNP) uses water in the presence of an ultrasound-assisted mechanical method (UL_CNP). However, the use of UL_CNP for the synthesis and optimization of silver nanoparticles (AgNP), particularly their anticancer and photocatalytic properties, remains unexplored. Hence, this research aimed to employ a green method using UL_CNP and silver nitrate to produce AgNP (UL_AgNP) with a small size and assess its potential toxicity, anticancer, and photocatalytic activities. The synthesis condition was optimized using the Box-Behnken design method. The synthesized UL_AgNP showed the surface plasmon resonance peak at 458 nm. The optimized synthesis condition produced spherically shaped UL_AgNP with a size of 5.21 ± 1.92 nm and a zeta potential of -26.33 ± 0.93 mV. An X-ray diffraction analysis exhibited intense Bragg's reflection peaks at (111), (200), (220), and (311), having a face-centered cubic structure of AgNP. Attenuated total reflectance-Fourier-transform infrared spectroscopy and energy-dispersive X-ray spectroscopy further confirmed the presence of silver in the synthesized UL_AgNP. The brine shrimp lethality test of UL_AgNP reported a lethal concentration 50 value of <7.8 µg/mL after 24 h. The UL_AgNP exhibited antiproliferative activity against MCF-7 cells with a half-maximal inhibitory concentration value of 4.96 ± 0.31 µg/mL by inducing S-phase cell cycle arrest, apoptotic effect, and reduction of cell migration. Furthermore, UL_AgNP proved its efficient photocatalytic activity against methylene blue dye (50.22 % ± 0.06 %, after 10 min at a concentration of 50 µg/mL). Therefore, the UL_AgNP exhibited promising antiproliferative activity against MCF-7 cells, highlighting their potential as a therapeutic agent. Further investigations are needed to elucidate the precise mechanism of their action.

In vivo visualization of environmentally relevant microplastics and evaluation of gut barrier damages in Artemia franciscana.

Although irregularly-shaped label-free microplastics (MPs) are predominantly distributed in the environment, non-destructive analysis of environmentally relevant MPs in organisms is still challenging. The purpose of the study is to suggest in vivo visual evidence of the uptake and effect of environmentally relevant MPs in organism. Transparent irregularly-shaped high-density polyethylene was selected as an environmentally relevant model MP and exposed to brine shrimp (Artemia franciscana). As a result, we suggest the application of SEM/EDX and coherent anti-Stokes Raman scattering (CARS) microspectroscopy as complementary tools to secure in vivo visual evidence of irregularly-shaped unlabeled MPs in living organisms without chemical digestion for biodistribution observations. Biological transmission electron microscopy also provides how ingested MPs physically affects the digestive tract in the brine shrimp which is rarely reported. In terms of environmental implications, this study would advance ecotoxicological research on microplastic pollution by providing a cutting-edge tool for investigating the bioavailability and ecotoxicity of environmentally relevant MPs in ecosystems.

Induction and comparative resuscitation of viable but nonculturable state on Vibrio parahaemolyticus serotypes O3:K6 and O1:K25.

Vibrio parahaemolyticus, an important food-borne pathogens found to be associated with seafoods and marine environs. It has been a topic of debate for many decades that most pathogens are known to enter a viable but nonculturable (VBNC) state under cold temperature and nutrient limited conditions. The present study examined the time required for the induction of VBNC state and the revival strategies of both the endemic O3:K6 and O1:K25 sporadic strains of V. parahaemolyticus. The results revealed that V. parahaemolyticus survived even after 55 days of incubation in nutrient starved media such as phosphate buffered saline (PBS) and Coastal Water (CW) and could be recovered by temperature upshift method, and compared the resuscitation using Dulbecco's Modified Eagle Medium (DMEM), sheep blood serum, chitin flakes with live Artemia salina, and the results suggests that chitin plays a significant role in regulating the VBNC state. It was also confirmed by Confocal Laser Scanning Microscopy (CLSM) and Scanning Electron Microscope (SEM) analysis that VBNC cells can alter their morphology to coccoid forms in order to survive in most extreme nutrient limited environment. Further data on the promoting factors and the exact mechanism that resuscitate VBNC V. parahaemolyticus in cold natural environments and frozen foods are needed to perform a robust risk assessment.

Rearing condition influences gene expression in postlarval American lobster (Homarus americanus).

The American lobster (Homarus americanus) is an economically important species in the western Atlantic and its climate-driven range shift northward along the east coast of the United States is well documented. The thermal tolerance of lab-reared postlarvae of this species has been extensively investigated to better understand settlement and recruitment dynamics. However, there have been few studies focused on wild-caught postlarvae, and even fewer that have analyzed lab-rearing conditions in context of diet. In this study, we investigated gene transcriptional changes in postlarvae caught in the wild, as well as postlarvae reared in the laboratory on a brine shrimp diet or a wild-sourced zooplankton diet. We found between wild-caught and brine shrimp-reared larvae 3,682 differentially expressed genes, and between wild and zooplankton-reared postlarvae 3,939 differentially expressed genes. Between the two lab-reared groups fed different diets 2,603 genes were differentially expressed. We also exposed individuals in all rearing groups to chronic temperature treatments of 8°C and 26°C and found that both temperature extremes elicit 68-95% fewer transcriptional changes in wild postlarvae compared to either lab-reared group. In wild postlarvae, we identified differential expression of transcripts within the FoxO signaling pathway, a signaling pathway with a central role in cellular physiology, as potential molecular markers for cold tolerance in the American lobster. These findings contextualize the current literature on lab-reared postlarvae as containing conservative estimates for in situ organisms and can be used to inform population distribution modeling efforts. They also provide evidence for the need to adjust lab-rearing techniques or source wild larval crustaceans to augment studies of larval biology.

Analysis of Chemical Composition, Antioxidant Activity, and Toxicity of Essential Oil from Virola sebifera Aubl (Myristicaceae).

Volatile oils or essential oils (EOs) were extracted from three V. sebifera samples (labeled as A, B, and C) in September 2018 and February 2019; the extraction process involved hydrodistillation of the leaves. The chemical compositions of the EOs were analyzed using gas chromatography-mass spectrometry (GC/MS). The volatile components were identified by comparing their retention indices and mass spectra with standard substances documented in the literature (ADAMS). The antioxidant activity of the EOs was evaluated using 2, 2-diphenyl-1-picrylhydrazyl (DPPH), while their toxicity was assessed using Artemia salina Leach. Molecular docking was utilized to examine the interaction between the major constituents of V. sebifera EO and acetylcholinesterase (AChE), a molecular target linked to toxicity in A. salina models. The EO obtained from specimen A, collected in September 2018, was characterized by being primarily composed of (E,E)-α-farnesene (47.57%), (E)-caryophyllene (12.26%), and α-pinene (6.93%). Conversely, the EO from specimen A, collected in February 2019, was predominantly composed of (E,E)-α-farnesene (42.82%), (E)-caryophyllene (16.02%), and bicyclogermacrene (8.85%), the EO from specimen B, collected in September 2018, primarily contained (E,E)-α-farnesene (47.65%), (E)-caryophyllene (19.67%), and α-pinene (11.95%), and the EO from the leaves collected in February 2019 was characterized by (E,E)-α-farnesene (23.57%), (E)-caryophyllene (19.34%), and germacrene D (7.33%). The EO from the leaves collected in September 2018 contained (E,E)-α-farnesene (26.65%), (E)-caryophyllene (15.7%), and germacrene D (7.72%), while the EO from the leaves collected in February 2019 was primarily characterized by (E,E)-α-farnesene (37.43%), (E)-caryophyllene (21.4%), and α-pinene (16.91%). Among these EOs, sample B collected in February 2019 demonstrated the highest potential for inhibiting free radicals, with an inhibition rate of 34.74%. Conversely, the EOs from specimen A exhibited the highest toxic potentials, with an lethal concentration 50 (LC50) value of 57.62 ± 1.53 µg/mL, while specimen B had an LC50 value of 74.72 ± 2.86 µg/mL. Molecular docking results suggested that hydrophobic interactions significantly contributed to the binding of the major compounds in the EO from sample B to the binding pocket of AChE.