Alien plant species distribution in Romania: a nationwide survey following the implementation of the EU Regulation on Invasive Alien Species.

Background: Biological invasions pose an increasing risk to nature, social security and the economy, being ranked amongst the top five threats to biodiversity. Managing alien and invasive species is a priority for the European Union, as outlined in the EU Biodiversity Strategy for 2030 and the Kunming-Montreal Global Biodiversity Framework. Alien plant species are acknowledged to impact the economy and biodiversity; thus, analysing the distribution of such species provides valuable inputs for the management and decision-making processes. The database presented in the current study is the first consolidated checklist of alien plant species that are present in Romania, both of European Union concern and of national interest. This database complements a prior published distribution, based only on records from literature, bringing new information regarding the occurrence of alien plants in Romania, as revealed by a nationwide field survey. We consider this database a valuable instrument for managing biological invasions at both national and regional levels, as it can be utilised in further research studies and in drafting management and action plans, assisting stakeholders in making informed decisions and implementing management actions. New information: We present the results of the first nationwide survey of alien plant species in Romania, conducted between 2019 and 2022, in the framework of a national project coordinated by the Ministry of Environment, Waters and Forests and the University of Bucharest. The present database complements and updates the database published by Sirbu et. al (2022), which included occurrence records published until 2019. The new database includes 98323 occurrence records for 396 alien plant species in 77 families, with most species belonging to the Asteraceae family. One alien plant species in our database, the black locust Robiniapseudoacacia L., had more than 10,000 occurrence records. The distribution database also includes information on newly-reported invasive alien plant species of European Union concern in Romania (i.e. the floating primrose-willow Ludwigiapeploides (Kunth) P.H.Raven) and documents the presence of plants in 44 additional families compared to Sirbu et al. (2022). Each entry includes information on species taxonomy, location, year, person who recorded and identified the alien plant, geographical coordinates and taxon rank.

Evaluation of the Morphological, Physiological and Biochemical Effects Induced by Coragen 20 SC in Some Non-Target Species.

Coragen 20 SC is an insecticide based on chlorantraniliprole that is applied on many crops. Considered an effective product with an incremental cost-benefit ratio, it has been widely used globally. Residual pesticides affect non-target organisms, so it is necessary to explore the possible effects induced by these xenobiotics on different species. This work aimed to assess some morphological, physiological and biochemical effects induced by Coragen 20 SC on two non-target species: Perca fluviatilis (Linné, 1758) and Triticum aestivum L. The concentrations used were the same for all tested species (0.0125, 0.025 and 0.05 mL L-1), and the experiments were of the acute, subchronic and chronic type. The toxicological effects of Coragen 20 SC on perch recorded behavioral changes, a decrease in respiratory rate and oxygen consumption, an increase in blood glucose levels and a decrease in the number of erythrocytes and leukocytes. The results obtained from the evaluation of Coragen 20 SC toxicity using the Triticum test indicate a weak to moderate phytotoxicity for the considered parameters at the applied doses. Only the assimilatory pigments were significantly modified at the concentration of 0.025 mL L-1 for the growth of the axial organs and the wet and dry weight, with the changes obtained not being statistically significant.

Ecotoxicological Risk Assessment of Actellic 50 EC Insecticide on Non-Target Organisms in Parallel with the Application of Standardized Tests.

This paper contributes to the ecotoxicological risk assessment of the Actellic 50 EC insecticide (with 50% pirimiphos-methyl as the active substance) tested on non-target organisms. The insecticide concentrations tested were the same for all organisms (0.1, 0.01, and 0.001 mg L-1 of Actellic 50 EC), with an exposure of 3-5-21 days for plants and 4-5-14 days for animals. The non-target organisms tested were both plants (wheat and two ferns) and animals (the Prussian carp and marsh frog tadpoles). The tested insecticide significantly inhibited the growth of roots in wheat, a result that was also confirmed by a microbiotest application (62% root growth inhibition in sorghum and 100% germination inhibition in white mustard and garden cress). In ferns, even for the lowest concentration, the percentage of germinated spores was inhibited by 40% for Asplenium scolopendrium. The recorded toxicological effects of Actellic 50 EC upon the Prussian carp included a decrease in the respiratory rate and oxygen consumption, an increase in the number of erythrocytes and leukocytes, and an increase in blood glucose levels. The highest concentration (0.1 mg L-1 of Actellic 50 EC) caused a 50% decrease in the survival rate of marsh frog tadpoles after 5 days of exposure, negatively affecting body volume and length. Given the high degree of toxicity of the insecticide Actellic 50 EC, we recommend continuing investigations on non-target species, including both plants and animals, as the sub-chronic effects are quite little known in the scientific literature.

Grapevine Plants Management Using Natural Extracts and Phytosynthesized Silver Nanoparticles.

Starting from the well-known antimicrobial properties of silver nanoparticles, the goal of this study is to evaluate the influence of two "green" recipes, namely an alcoholic extract of Dryopteris filix-mas (L.) Schott and a dispersion of silver nanoparticles phytosynthesized using the extract on grapevine pathogens. The influence of some grapevine parameters (pith/wood rapport, soluble sugars, starch, total sugars, total water content, length of young shoots, number of grapes) in field experiments was also studied. The study was conducted on four clones (Feteasca alba 97 St., Feteasca neagra 6 St., Feteasca regala 72 St., and Cabernet Sauvignon 131 St.) located in vegetation pots inside a greenhouse. For the phytosynthesis of the silver nanoparticles (AgNPs) we used a scaled-up technology, allowing us to obtain large quantities of nanoparticles-containing solution. The AgNPs analysis by X-ray diffraction and transmission electron microscopy confirmed the synthesis of spherical and quasi-spherical nanoparticles of 17 nm average diameter and 6.72 nm crystallite size. The field experiments registered different responses of the four clones to the treatment, using both the natural extracts and phytosynthesized nanoparticles solution. Both recipes exhibited a protective effect against the Uncinula necator pathogen. For the treatment using phytosynthesized nanoparticles, significant increases in the pith/wood ratio for white wine clones (Feteasca alba 97 St. and Feteasca regala 72 St.) were observed. The biochemical analyses revealed other significant increases of soluble sugars (red wine clones-Feteasca neagra and Cabernet Sauvignon/second year), starch (Feteasca alba and Cabernet Sauvignon in 2021 for both clones), total sugars (Feteasca alba and Feteasca neagra in 2021 for both clones), and of total water content (Feteasca alba and Feteasca neagra in 2021 for both clones), respectively. The applied treatments also led to an increase of young shoots length and grape numbers for all clones as compared to the control (chemical pesticide), which would suggest a potential biostimulant effect of the recipes.

Ecotoxicological Studies on the Action of Actara 25 WG Insecticide on Prussian Carp (Carassius gibelio) and Marsh Frog (Pelophylax ridibundus).

The toxic action of the Actara 25 WG insecticide (it contains 25% thiamethoxam as an active substance) in non-lethal doses was studied in two species of aquatic organisms-the Prussian carp (Carassius gibelio) and the marsh frog (Pelophylax ridibundus)-at two thermal levels, 6-8 °C (low temperature) and 18-20 °C (room temperature), respectively. In the Prussian carp, we recorded decreases in oxygen consumption and stimulation of the respiratory rhythm, changes that were more pronounced in the case of intoxicated fish and when the species were kept at room temperature. The histopathology of the lung in the frog illustrated the thickening of the conjunctival septum, an increase in the number of mucous cells, and an increase in the ratio between the diameter of the nucleus and the diameter of the pneumocyte. All of these changes were more pronounced in the animals kept at higher temperature. Our study looks at the extent to which temperature changes can influence the ability of poikilothermic organisms to withstand the presence of toxic substances in the environment as a result of the impact of the use of insecticides in agriculture. The two tested organisms are a common presence for the study area, which was affected in the last decade by climate change.

Hericium erinaceus (Bull.) Pers. Ethanolic Extract with Antioxidant Properties on Scopolamine-Induced Memory Deficits in a Zebrafish Model of Cognitive Impairment.

Hericium erinaceus (H. erinaceus) is a rare and appreciated fungal species belonging to the division Basidiomycota used for centuries in traditional Chinese medicine for its medicinal value. This species of mushrooms brings the most diverse benefits for the human body, and can have beneficial effects for treating Alzheimer's disease (AD). This study investigated whether ethanolic extract from the fungal biomass of H. erinaceus enhances cognitive function via the action on cholinergic neurons using the scopolamine (SCOP)-induced zebrafish (Danio rerio) model of memory impairment. The ethanolic extract from the fungal biomass of H. erinaceus was previously obtained using an ultrasonic extraction method (UE). The administration of H. erinaceus extract to zebrafish, with a pattern of AD induced by scopolamine, showed an improvement in memory evaluated by behavioral and biochemical tests on brain tissue. These results suggest that H. erinaceus has preventive and therapeutic potentials in managing memory deficits and brain oxidative stress in zebrafish with AD.

Effects of the Hydroethanolic Extract of Lycopodium selago L. on Scopolamine-Induced Memory Deficits in Zebrafish.

This scientific research focused on the production of hydroethanolic extract of the plant species Lycopodium selago L. (L. selago) by the ultrasound-assisted extraction (USAE) and the identification of biocompounds with high antioxidant activity is of interest for possible phytotherapeutic treatment against Alzheimer's disease (AD). The extract was phytochemically analyzed to investigate polyphenols, flavonoids, and identify the sesquiterpenoid alkaloid huperzine A (HupA), which is known in the literature for its great relevance in AD. Evaluation and comparison of the antioxidant activity of the extract were performed by four complementary spectrophotometric methods (DPPH, FRAP, ABTS, ORAC). In vitro tests of the extract showed an excellent reciprocal link between the concentration of polyphenols and the measurement of the antioxidant activity of the extract with the sesquiterpenoid HupA. To confirm the antioxidant activity, L. selago hydroethanolic extract was administered in vivo to zebrafish (Danio rerio) with a pattern of scopolamine-induced cognitive impairment. Moreover, this study explored a possible correlation between the expression of oxidative stress markers in the brain tissue with the behavior of the scopolamine zebrafish model. In vivo tests showed that this fern could be used as a nutritional supply and as a phytotherapeutic method to prevent or treat various neurodegenerative diseases that call for high-nutritive-value medications.

Application of Polypodiopsida Class in Nanotechnology-Potential towards Development of More Effective Bioactive Solutions.

The area of phytosynthesized nanomaterials is rapidly developing, with numerous studies being published yearly. The use of plant extracts is an alternative method to reduce the toxic potential of the nanomaterials and the interest in obtaining phytosynthesized nanoparticles is usually directed towards accessible and common plant species, ferns not being explored to their real potential in this field. The developed nanoparticles could benefit from their superior antimicrobial and antioxidant properties (compared with the nanoparticles obtained by other routes), thus proposing an important alternative against health care-associated and drug-resistant infections, as well as in other types of applications. The present review aims to summarize the explored application of ferns in nanotechnology and related areas, as well as the current bottlenecks and future perspectives, as emerging from the literature data.

Optimization of Ultrasonic Extraction to Obtain Erinacine A and Polyphenols with Antioxidant Activity from the Fungal Biomass of Hericium erinaceus.

Hericium erinaceus is a medicinal fungal species that produces the active biological metabolite erinacine A with strong antioxidant activity. The classical extraction techniques used to date to obtain metabolites from this fungal species require high consumption of resources and energy and, in the end, prove to be expensive and inefficient, especially on a biomedical scale. The aim of this research is based on the development of an ultrasonic extraction (UE) method for the identification and extraction of biological compounds with high antioxidant activity from the mycelia of H. erinaceus biomass developed through a solid cultivation process. The extraction process was optimized by varying parameters to determine the best extraction yield of metabolites involved in such antioxidant activity, using the response surface methodology (RSM). The physicochemical analyses were oriented towards the investigation of polyphenols, flavonoids, and the diterpenoid erinacine A. It is highlighted that there is a very good mutual connection between the concentration of polyphenols and flavonoids in the extracts studied and the diterpenoid erinacine A. Also, this study describes an efficient and qualitative extraction method for extracting natural antioxidants from the H. erinaceus mushroom, since toxic solvents were not used in the developed extraction procedure. This biomass can be used both as a food source and as a possible phytotherapeutic tool in the prevention or treatment of various neurodegenerative disorders that require drugs with strong antioxidant activity.

Phytosynthesis of gold and silver nanoparticles enhance in vitro antioxidant and mitostimulatory activity of Aconitum toxicum Reichenb. rhizomes alcoholic extracts.

Extracts obtained from different plant species proved to be a valuable tool in various biomedical applications. In the same time, the phytosynthesis of noble metal nanoparticles represents an already well-established route for obtaining nanoparticles with biological activity. The present paper studies the antioxidant activity and the cytogenetic effects of the alcoholic extracts from rhizomes of Aconitum toxicum Rchb., before and after the phytosynthesis of gold and silver nanoparticles, on the meristematic root cells of Allium cepa L., and on the general mitotic index and the progression rate through the mitotic phases, respectively, as well as on the genetic material organized in chromosomes. The extracts were characterized in terms of total polyphenolics content (1.49% and, respectively, 2.29%) and aconitine content (by HPLC - 4.891 mg/L and, respectively, 18.211 mg/L), while the phytosynthesis of metallic nanoparticles was monitored by UV-Vis spectrometry, X-ray diffraction, X-ray fluorescence and electron microscopy. Both the extracts and the obtained nanoparticles were evaluated for antioxidant potential (the antioxidant activity ranging between 78% and 84.32%) and cytogenetic effects. The obtained results prove the phytosynthesis of AgNPs and AuNPs with dimensions ranging from 9 nm to 15 nm for AuNPs, respectively from 53 nm to 67 nm for AgNPs. The extracts obtained from rhizomes of A. toxicum Rchb. induced mitotic stress, as well as a series of nuclear and mitotic aberrations. The biosynthesis of AgNPs and AuNPs intensified the antioxidant and mitostimulatory activity of the extracts.

Lead-Induced Physiological, Biochemical and Enzymatic Changes in Asplenium scolopendrium L.

The paper aims to determine the lead-induced physiological, biochemical and enzymatic changes in Asplenium scolopendrium, which could represent biomarkers used in environmental assessment. Of all the physiological processes, photosynthesis and respiration were analyzed and the enzymatic and biochemical determinations focused on catalase activity, assimilatory pigment concentration, polyphenol content and lead presence in tissues.The stress induced by the exposure to Pb of the species Asplenium scolopendrium determined an increase in the carotenoid content, the catalase activity, the total polyphenol content and also enhanced the respiration potential. No significant changes were recorded regarding the chlorophyll content and the photosynthetic activity. The recorded changes may be used as non-specific markers in the assessment of the impact of Pb on plants (Asplenium scolopendrium).

The Influence of Six Pesticides on Physiological Indices of Pelophylax Ridibundus (Pallas, 1771).

The objective of the study is to screen for morphological, biochemical and histological changes induced by six widely used pesticides (Reldan 40EC, Actara 25WG, Tilt 250EC, Champion 50WG, Fusilade Forte, Dual Gold 960EC) in the amphibian species Pelophylax ridibundus (Pallas, 1771). Highly degenerative changes were observed in animals cultured at 22-24°C, compared to those cultured at 4-6°C. The hepatosomatic index increased upon exposure to almost all of the pesticides, the erythrocyte number decreased upon exposure to all pesticides except Reldan 40EC, while leucopenia was observed only for Reldan 40EC and Actara 25WG. Hyperglycemia was observed upon administration of pesticides (except Champion 50WG and Fusilade Forte, for which hypoglycemia is registered), while a decrease in cholesterol levels was induced by nearly all pesticides. Triglycerides varied only slightly. The results suggest that chronic pesticides exposure can lead to alteration of various indices, as well as to hepatic lesions in amphibians.

The Protective Role of Thiourea on Leuciscus cephalus Exposed to Sublethal Doses of Pendigan 330EC (Pendimethalin) Herbicide.

The aim of the study was to investigate the protective role of thiourea on the physiological, hematological, biochemical and histopathological parameters of Leuciscus cephalus exposed to sublethal concentration of Pendigan 330 EC herbicide. The animals were divided in four experimental groups (control, animals subjected to 1 ‰ thiourea, animals subjected to 4 × 10(-4) mL/L herbicide and, respectively, animals subjected to 4 × 10(-4) mL/L herbicide and 1 ‰ thiourea). Exposure of European chub to herbicide administered in water for 2 weeks determined installation of pathological changes in the liver and gills tissues. Also, were observed a decrease in the number of white blood cells and oxygen consumption, breathing frequency, and an increase in the number of red blood cells and glycaemia values. Thiourea counteracts the toxic action, describing itself as normal liver parenchyma and normal gills in animals intoxicated with herbicide, without lesion, and a return to normal values of the studied markers.