Essential Oils as Nematicides in Plant Protection-A Review.

By 2030, the European Commission intends to halve chemical pesticide use and its consequent risks. Among pesticides, nematicides are chemical agents used to control parasitic roundworms in agriculture. In recent decades, researchers have been looking for more sustainable alternatives with the same effectiveness but a limited impact on the environment and ecosystems. Essential oils (EOs) are similar bioactive compounds and potential substitutes. Different studies on the use of EOs as nematicides are available in the Scopus database in the scientific literature. These works show a wider exploration of EO effects in vitro than in vivo on different nematode populations. Nevertheless, a review of which EOs have been used on different target nematodes, and how, is still not available. The aim of this paper is to explore the extent of EO testing on nematodes and which of them have nematicidal effects (e.g., mortality, effects on motility, inhibition of egg production). Particularly, the review aims to identify which EOs have been used the most, on which nematodes, and which formulations have been applied. This study provides an overview of the available reports and data to date, downloaded from Scopus, through (a) network maps created by VOSviewer software (version 1.6.8, Nees Jan van Eck and Ludo Waltman, Leiden, The Netherlands) and (b) a systematic analysis of all scientific papers. VOSviewer created maps with keywords derived from co-occurrence analysis to understand the main keywords used and the countries and journals which have published most on the topic, while the systematic analysis investigated all the documents downloaded. The main goal is to offer a comprehensive understanding of the potential use of EOs in agriculture as well as which directions future research should move toward.

Taxonomic and functional nematode diversity in Maldivian coral degradation zones: patterns across reef typologies and depths.

The intensification of dredging and infilling activities in lagoons and on coral reefs are common practices in the Maldivian archipelago, and these activities alter the biodiversity of the bioconstructors and the functioning of the ecosystem. The alteration of environmental factors can also affect inconspicuous fauna, such as free-living nematodes. The implications of a reduction in biodiversity may transcend decreased taxonomic diversity, resulting in changes in functional diversity and redundancy; however, how the environmental conditions and human pressure affects the functionality of nematodes in Maldivian coral degradation zones (CDZs) remain poorly understood. In this paper, we examined changes in the taxonomic and functional diversity and the functional redundancy in nematode communities regarding the geographic location (atolls with various levels of human pressure), the exposure and topography of the reef (lagoon and ocean), the slope of the reef, and the depth. The functional diversity and redundancy were evaluated by considering two main biological traits of nematodes: i) the trophic strategy, and ii) the life strategy. The extremely high number of nematode genera observed in the Maldives is supported by the high complexity of the carbonate sediments. The reef exposure and depth were the most relevant environmental factors that influenced the taxonomic and functional diversity. The functional diversity, according to the trophic strategies, mirrored the taxonomic diversity because the adaptive plasticity of nematode buccal cavity structures is closely associated with the high biodiversity of the phylum. The high abundance of k-strategists in ocean reefs may indicate a higher ecological quality when compared to lagoon reefs; however, the absence of significant differences in life strategy functional diversity and functional redundancy indicates that a recovery process is underway. Analyses of nematode communities should be combined with standard investigations of reef bioconstructors during monitoring activities to assess the vulnerability of CDZ systems to future disturbances and facilitate the adoption of the most appropriate preventative actions.

Antioxidant and Anti-Inflammaging Ability of Prune (Prunus Spinosa L.) Extract Result in Improved Wound Healing Efficacy.

Prunus spinosa L. fruit (PSF) ethanol extract, showing a peculiar content of biologically active molecules (polyphenols), was investigated for its wound healing capacity, a typical feature that declines during aging and is negatively affected by the persistence of inflammation and oxidative stress. To this aim, first, PSF anti-inflammatory properties were tested on young and senescent LPS-treated human umbilical vein endothelial cells (HUVECs). As a result, PSF treatment increased miR-146a and decreased IRAK-1 and IL-6 expression levels. In addition, the PSF antioxidant effect was validated in vitro with DPPH assay and confirmed by in vivo treatments in C. elegans. Our findings showed beneficial effects on worms' lifespan and healthspan with positive outcomes on longevity markers (i.e., miR-124 upregulation and miR-39 downregulation) as well. The PSF effect on wound healing was tested using the same cells and experimental conditions employed to investigate PSF antioxidant and anti-inflammaging ability. PSF treatment resulted in a significant improvement of wound healing closure (ca. 70%), through cell migration, both in young and older cells, associated to a downregulation of inflammation markers. In conclusion, PSF extract antioxidant and anti-inflammaging abilities result in improved wound healing capacity, thus suggesting that PSF might be helpful to improve the quality of life for its beneficial health effects.

Prunus spinosa Extract Loaded in Biomimetic Nanoparticles Evokes In Vitro Anti-Inflammatory and Wound Healing Activities.

Prunus spinosa fruits (PSF) contain different phenolic compounds showing antioxidant and anti-inflammatory activities. Innovative drug delivery systems such as biomimetic nanoparticles could improve the activity of PSF extract by promoting (i) the protection of payload into the lipidic bilayer, (ii) increased accumulation to the diseased tissue due to specific targeting properties, (iii) improved biocompatibility, (iv) low toxicity and increased bioavailability. Using membrane proteins extracted from human monocyte cell line THP-1 cells and a mixture of phospholipids, we formulated two types of PSF-extract-loaded biomimetic vesicles differing from each other for the presence of either 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) or 1,2-dioleoyl-sn-glycero-3-phospho-(1'-rac-glycerol) (DOPG). The biological activity of free extract (PSF), compared to both types of extract-loaded vesicles (PSF-DOPCs and PSF-DOPGs) and empty vesicles (DOPCs and DOPGs), was evaluated in vitro on HUVEC cells. PSF-DOPCs showed preferential incorporation of the extract. When enriched into the nanovesicles, the extract showed a significantly increased anti-inflammatory activity, and a pronounced wound-healing effect (with PSF-DOPCs more efficient than PSF-DOPG) compared to free PSF. This innovative drug delivery system, combining nutraceutical active ingredients into a biomimetic formulation, represents a possible adjuvant therapy for the treatment of wound healing. This nanoplatform could be useful for the encapsulation/enrichment of other nutraceutical products with short stability and low bioavailability.

Development of long circulating magnetic particle imaging tracers: use of novel magnetic nanoparticles and entrapment into human erythrocytes.

Aim: Magnetic particle imaging (MPI) is highly promising for biomedical applications, but optimal tracers for MPI, namely superparamagnetic iron oxide-based contrast agents, are still lacking. Materials & methods: The encapsulation of commercially available nanoparticles, specifically synomag®-D and perimag®, into human red blood cells (RBCs) was performed by a hypotonic dialysis and isotonic resealing procedure. The amounts of superparamagnetic iron oxide incorporated into RBCs were determined by Fe quantification using nuclear magnetic resonance and magnetic particle spectroscopy. Results: Perimag-COOH nanoparticles were identified as the best nanomaterial for encapsulation in RBCs. Perimag-COOH-loaded RBCs proved to be viable cells showing a good magnetic particle spectroscopy performance, while the magnetic signal of synomag-D-COOH-loaded RBCs dropped sharply. Conclusion: Perimag-COOH-loaded RBCs could be a potential tool for MPI diagnostic applications.

The curious and neglected soft-bodied meiofauna: Rouphozoa (Gastrotricha and Platyhelminthes).

Gastrotricha and Platyhelminthes form a clade called Rouphozoa. Representatives of both taxa are main components of meiofaunal communities, but their role in the trophic ecology of marine and freshwater communities is not sufficiently studied. Traditional collection methods for meiofauna are optimized for Ecdysozoa, and include the use of fixatives or flotation techniques that are unsuitable for the preservation and identification of soft-bodied meiofauna. As a result, rouphozoans are usually underestimated in conventional biodiversity surveys and ecological studies. Here, we give an updated outline of their diversity and taxonomy, with some phylogenetic considerations. We describe successfully tested techniques for their recovery and study, and emphasize current knowledge on the ecology, distribution and dispersal of freshwater gastrotrichs and microturbellarians. We also discuss the opportunities and pitfalls of (meta)barcoding studies as a means of overcoming the taxonomic impediment. Finally, we discuss the importance of rouphozoans in aquatic ecosystems and provide future research directions to fill in crucial gaps in the biology of these organisms needed for understanding their basic role in the ecology of benthos and their place in the trophic networks linking micro-, meio- and macrofauna of freshwater ecosystems.

Secosterol-B affects endoplasmic reticulum structure in endothelial cells.

Oxysterols, oxidized derivatives of cholesterol found in LDL and atherosclerotic plaques, trigger several biological responses involved in the initiation and progression of atherosclerosis. Endothelial dysfunction, which occurs when vascular homeostasis is altered, plays a key role in the pathogenesis of several metabolic diseases. The contribution of endoplasmic reticulum (ER) stress to endothelial disfunction is a relatively recent area of investigation. There is a well-established link between LDL oxidation and ER stress but the role played by specific products of lipid oxidation into this interaction is still to be defined. The present study shows that secosterol-B (SEC-B), 3ß-hydroxy-5ß-hydroxy-B-norcholestane-6ßcarboxaldehyde, a cholesterol autoxidation product recently identified in the atherosclerotic plaque, is able to induce ER stress in HUVEC cells, as revealed by significant expansion and change of structure. At low doses, i.e. 1 and 5 µM, cells try to cope with this stress by activating autophagy and the ubiquitin proteasome system in the attempt to restore ER function. However, at higher doses, i.e. 20 µM, cell apoptosis occurs in a pathway that involves early phosphorylation of eIF2α and NF-kB activation, suggesting that the adaptive program fails and the cell activates the apoptotic program. These findings provide additional insight about the role of oxysterols in endothelial dysfunction and its potential involvement in atherosclerotic pathophysiology.

Bioeffects of Prunus spinosa L. fruit ethanol extract on reproduction and phenotypic plasticity of Trichoplax adhaerens Schulze, 1883 (Placozoa).

The aim of this work was to test and analyse the bioeffects of Prunus spinosa L. (Rosacaee) fruit ethanol extract on Trichoplax adhaerens Schulze, 1883 (Placozoa) laboratory cultures which-for the first time-were employed as in vivo biological model to assess the bioactivity of a natural extract. The ethanol extract of P. spinosa was administrated during a 46 day experimental period; ultrastructural (by optical, confocal, TEM and SEM microscopy) and morphometric analyses indicated that treated Trichoplax adhaerens showed significant differences in viability, reproductive modalities, body shape and colour with respect to the control group. Finally, P. spinosa bioactive compounds seem to exert profound protective effects on T. adhaerens reproduction and phenotype. Our results may support additional investigations related to other bioactive compounds properties useful for nutraceutical preparations to be used as food supplements.

Integrative taxonomy of a new Redudasys species (Gastrotricha: Macrodasyida) sheds light on the invasion of fresh water habitats by macrodasyids.

The order Macrodasyida (Gastrotricha) includes over 350 marine species, and only 3 freshwater species (Marinellina flagellata, Redudasys fornerise, R. neotemperatus). Herein we describe a new freshwater species of Macrodasyida, Redudasys brasiliensis sp. nov., from Brazil through an integrative taxonomic approach. The external morphology and internal anatomy were investigated using differential interference contrast microscopy, confocal microscopy, scanning and transmission electron microscopy. The systematization of the new taxon was inferred by nuclear (18S and 28S) and mitochondrial (COI) genes, and its intra-order relationships were assessed using data from most of available macrodasyids. Phylogenetic analyses yielded congruent trees, in which the new taxon is nested within the family Redudasyidae, but it was genetically distinct from the other species of the genus Redudasys. The new species shares the gross morphology and reproductive traits with other Redudasyidae and the presence of only 1 anterior adhesive tube per side with Redudasys neotemperatus, but it has a specific pattern of ventral ciliation and muscle organization. Results support the hypothesis that dispersion into fresh water habitats by Macrodasyida and Chaetonotida taxa occurred independently and that within Macrodasyida a single lineage invaded the freshwater environment only once. Furthermore, the Neotropical region seems to be peculiar for the evolution of the freshwater macrodasyid clade.

Some epibiont suctorian ciliates from meiofaunal organisms of Maldivian archipelago with description of a new ciliate species.

Five species of suctorian ciliates belonging to four different genera attached to meiofaunal organisms of Maldivian archipelago (Suvadiva Atoll) were documented. Three rare species (Acineta sp., Thecacineta urceolata Liao & Dovgal, 2015 and Limnoricus ceter Jankowski, 1981) and one common suctorian ciliate Thecacineta calix (Schröder, 1907) are re-described from nematodes (Croconema cinctum Cobb, 1920, Desmodorella tenuispiculum Allgén, 1928 and Desmodora sp.) and harpacticoid copepod basibionts. In addition, Trophogemma colantonii n. sp. colonizing the abdomen and legs of the harpacticoid copepod Stenhelia sp. is described. The new species differs from the relative species for the less flattened cell body, the absence of both lateral bundles of tentacles and separate lateral tentacles as well as for the presence of longitudinal ribs along the cuticle. The data obtained in this survey give not only new important information on the taxonomy and distribution of Suctorea, but also new insights on their relationship with meiofaunal organisms in one of the most diverse and productive marine micro-habitat of carbonate sediments.

Fauna Europaea: Gastrotricha.

Fauna Europaea provides a public web-service with an index of scientific names (including important synonyms) of all living European land and freshwater animals, their geographical distribution at country level (up to the Urals, excluding the Caucasus region), and some additional information. The Fauna Europaea project covers about 230,000 taxonomic names, including 130,000 accepted species and 14,000 accepted subspecies, which is much more than the originally projected number of 100,000 species. This represents a huge effort by more than 400 contributing specialists throughout Europe and is a unique (standard) reference suitable for many users in science, government, industry, nature conservation and education. Gastrotricha are a meiobenthic phylum composed of 813 species known so far (2 orders, 17 families) of free-living microinvertebrates commonly present and actively moving on and into sediments of aquatic ecosystems, 339 of which live in fresh and brackish waters. The Fauna Europaea database includes 214 species of Chaetonotida (4 families) plus a single species of Macrodasyida incertae sedis. This paper deals with the 224 European freshwater species known so far, 9 of which, all of Chaetonotida, have been described subsequently and will be included in the next database version. Basic information on their biology and ecology are summarized, and a list of selected, main references is given. As a general conclusion the gastrotrich fauna from Europe is the best known compared with that of other continents, but shows some important gaps of knowledge in Eastern and Southern regions.

New insights into placozoan sexual reproduction and development.

Unraveling animal life cycles and embryonic development is basic to understanding animal biology and often sheds light on phylogenetic relationships. A key group for understanding the evolution of the Metazoa is the early branching phylum Placozoa, which has attracted rapidly increasing attention. Despite over a hundred years of placozoan research the life cycle of this enigmatic phylum remains unknown. Placozoa are a unique model system for which the nuclear genome was published before the basic biology (i.e. life cycle and development) has been unraveled. Four organismal studies have reported the development of oocytes and one genetic study has nourished the hypothesis of sexual reproduction in natural populations at least in the past. Here we report new observations on sexual reproduction and embryonic development in the Placozoa and support the hypothesis of current sexual reproduction. The regular observation of oocytes and expressed sperm markers provide support that placozoans reproduce sexually in the field. Using whole genome and EST sequences and additional cDNA cloning we identified five conserved sperm markers, characteristic for different stages in spermatogenesis. We also report details on the embryonic development up to a 128-cell stage and new ultrastructural features occurring during early development. These results suggest that sperm and oocyte generation and maturation occur in different placozoans and that clonal lineages reproduce bisexually in addition to the standard mode of vegetative reproduction. The sum of observations is best congruent with the hypothesis of a simple life cycle with an alternation of reproductive modes between bisexual and vegetative reproduction.

Ultrastructural analyses support different morphological lineages in the phylum Placozoa Grell, 1971.

The morphology and ultrastructure of 10 clonal placozoan lineages were studied. We scored several morphological characters at a cellular and intracellular level and identified a number of morphological differences among clones. Some differences appear clone specific and allow recognizing five distinct lineages based on morphological criteria only. These data will be crucial for a yet to be established placozoan systematics. Furthermore, we here describe three new diagnostic morphological characters for Placozoa: a new structure in the upper epithelium, called "concave disc," two distinct subpopulations of fiber cells, and especially small cells in the body margin. Besides the fiber cells appear to be arranged in several layers forming a complex, three-dimensional net not previously described. We also describe the marginal cells as the formerly suggested potential stem-cell type. The basic morphology is revised.

Glucose-6-phosphate dehydrogenase expression associated with NADPH-dependent reactions in cerebellar neurons.

This review describes the variation of glucose-6-phosphate dehydrogenase (G6PD) activity in the main neurons of the molecular and granular layers as well as in the deep nuclei of the cerebellum as observed so far by optical and electron microscopy studies. Light microscopy and semiquantitative microphotometry of histochemical staining showed that the highest G6PD activity was expressed by Purkinje cells and neurons of the deep cerebellar nuclei; the elements of the molecular layer showed a diffuse G6PD staining, while the granular layer displayed only scattered G6PD activity. Electron microscopy analysis showed that the basket and stellate cells, as well as the Golgi cells, have a remarkable G6PD activity, while in the granule cells the enzyme was barely detectable. The results show that cerebellar G6PD activity changes with different neuron types as a function of its role in sustaining NADPH dependent pathways in these cells.

Effects of thiocolchicine on axonal cytoskeleton of the rat peroneus nerve.

Thiocolchicine is a colchicine-derivative used in the therapy of some diseases and extensively studied in the field of oncological research as antimitotic agent. Here we studied the activity of thiocolchicine on the cytoskeleton of the peroneus nerve, performing a histological and ultrastructural analysis. We observed a decrease in mean myelinated fiber area in thiocolchicine-treated rats in comparison to controls; this was due to a decrease in mean axoplasm area, while myelin thickness was constant. In the ultrastructural analysis a decrease in microtubule density and an increase in neurofilaments were found; moreover, the myelinated fibers seemed to be more affected in comparison to the unmyelinated axons. These findings are in agreement with the capability of binding to microtubule skeleton shared by all the colchicinoids.