Uncovering developmental diversity in the field.

Many developmental biologists seldom leave the lab for research, relying instead on establishing colonies of traditional and emerging model systems. However, to fully understand the mechanisms and principles of development and evolution, including the role of ecology and the environment, it is important to study a diverse range of organisms in context. In this Perspective, we hear from five research teams from around the world about the importance and challenges of going into the field to study their organisms of interest. We also invite you to share your own fieldwork stories on the Node.

Chemical and Sensory Properties of Waffles Supplemented with Almond Skins.

Almonds are one of the most produced nuts worldwide and numerous studies have shown that they have nutritional and medicinal characteristics, which gives them the possibility of being applied in various products. However, several by-products are generated during their production, which have characteristics of interest but remain underutilised, namely, the almond skins. This work aimed to study samples of waffles supplemented with almond skins. The waffles were evaluated for their total polyphenol content, antioxidant capacity, total flavonoids, ortho-diphenols, soluble sugars, starch, texture, and colour. They were also sensorially evaluated using a panel of tasters specialised in this type of evaluation and a quantitative descriptive analysis test (QDA) sensory test. The results showed that the waffles with the highest levels of phenolic compounds as well as the highest antioxidant activity (by the ABTS, DPPH, and FRAP methods) were the waffles supplemented with 10% almond skin. The total phenol contents obtained for the prepared extracts varied between 0.127 mg GAE/g and 0.415 mg GAE/g, the flavonoid contents ranged from 0.067 mg CAE/g to 0.339 mg CAE/g and the ortho-diphenol contents varied between 0.163 mg ACE /g and 0.303 mg ACE/g. Regarding the quantification of soluble sugars, the values were presented in percentage of fresh weight, and ranged from 30.148 to 38.054%; regarding the quantification of starch, the percentages varied from 14.488 to 21.982%. Sensorially, we verified that the samples were statistically different in terms of the descriptors "colour", "roasted aroma", and "dissolubility", with a higher score in these descriptors for the waffles with 10% of almond skin. This process of obtaining waffles, which can be industrialised, is interesting from both a nutritional point of view and for the possibility of creating new, differentiated, and innovative products.

Myoanatomy of the velvet worm leg revealed by laboratory-based nanofocus X-ray source tomography.

X-ray computed tomography (CT) is a powerful noninvasive technique for investigating the inner structure of objects and organisms. However, the resolution of laboratory CT systems is typically limited to the micrometer range. In this paper, we present a table-top nanoCT system in conjunction with standard processing tools that is able to routinely reach resolutions down to 100 nm without using X-ray optics. We demonstrate its potential for biological investigations by imaging a walking appendage of Euperipatoides rowelli, a representative of Onychophora-an invertebrate group pivotal for understanding animal evolution. Comparative analyses proved that the nanoCT can depict the external morphology of the limb with an image quality similar to scanning electron microscopy, while simultaneously visualizing internal muscular structures at higher resolutions than confocal laser scanning microscopy. The obtained nanoCT data revealed hitherto unknown aspects of the onychophoran limb musculature, enabling the 3D reconstruction of individual muscle fibers, which was previously impossible using any laboratory-based imaging technique.

Application of chemometric tools for the comparison of volatile profile from raw and roasted regional and foreign almond cultivars (Prunus dulcis).

In almonds, volatile compounds are major contributors to flavour, being scarce the current knowledge about their volatile profile. Hence, this work intended to characterize the volatile profile, using headspace solid-phase microextraction and gas chromatography-mass spectrometry, in raw and roasted almond cultivars (regional cvs. Amendoão, Bonita, Casanova, Molar and Pegarinhos and foreign cvs. Ferragnès and Glorieta). Overall, 35 compounds were identified, with major chemical classes being alcohols and aldehydes. In raw fruits, benzaldehyde and 3-methyl-1-butanol were key compounds, with roasting changing volatile profiles, increasing release of compounds, with predominance of hexanal and benzaldehyde. Cultivars Glorieta and Molar didn't show significant increase in aldehyde content after roasting, which may indicate higher resistance to heat-caused oxidation. The use of linear discriminant analysis and principal components analysis permitted the recognition of patterns in the volatile profiles, that can be useful for cultivars identification. This work allowed the characterization and monitoring changes caused by roasting of volatile components of less studied almond cultivars, identifying some that can withstand roasting procedures with reduced formation of compounds associated with off-flavours.

Evaluation of contrasting techniques for X-ray imaging of velvet worms (Onychophora).

Non-invasive imaging techniques like X-ray computed tomography have become very popular in zoology, as they allow for simultaneous imaging of the internal and external morphology of organisms. Nevertheless, the effect of different staining approaches required for this method on samples lacking mineralized tissues, such as soft-bodied invertebrates, remains understudied. Herein, we used synchrotron radiation-based X-ray micro-computed tomography to compare the effects of commonly used contrasting approaches on onychophorans - soft-bodied invertebrates important for studying animal evolution. Representatives of Euperipatoides rowelli were stained with osmium tetroxide (vapour or solution), ruthenium red, phosphotungstic acid, or iodine. Unstained specimens were imaged using both standard attenuation-based and differential phase-contrast setups to simulate analyses with museum material. Our comparative qualitative analyses of several tissue types demonstrate that osmium tetroxide provides the best overall tissue contrast in onychophorans, whereas the remaining staining agents rather favour the visualisation of specific tissues and/or structures. Quantitative analyses using signal-to-noise ratio measurements show that the level of image noise may vary according to the staining agent and scanning medium selected. Furthermore, box-and-whisker plots revealed substantial overlap in grey values among structures in all datasets, suggesting that a combination of semiautomatic and manual segmentation of structures is required for comprehensive 3D reconstructions of Onychophora, irrespective of the approach selected. Our results show that X-ray micro-computed tomography is a promising technique for studying onychophorans and, despite the benefits and disadvantages of different staining agents for specific tissues/structures, this method retrieves informative data that may eventually help address evolutionary questions long associated with Onychophora.

Morphological, mechanical and antioxidant properties of Portuguese almond cultivars.

The aim of this study was to evaluate morphological (of fruit and kernel), mechanical (namely shell rupture force) and antioxidant properties (including phenolics and flavonoid content) of five Portuguese almond cultivars, comparing them with two commercial cultivars (Glorieta and Ferragnès). Of the analyzed traits, nut and kernel dimensions varied substantially and were used to describe cultivars. However, some traditional cultivars recorded similar (Pegarinhos), or even higher (Amendoão, Casanova and Refêgo) nut and kernel weight than commercial cultivars. Furthermore, shelling percentage of traditional cultivar (Bonita) was higher than commercial cultivars. Rupture force necessary to break fruits of all traditional cultivars was higher than commercial ones, and was correlated to nut weight cultivars. The phenolics, flavonoids content and antioxidants were higher for Casanova. Parameters like high kernel weight, low percentages of double kernels or losses during shelling and considerable higher phenolics and flavonoids content may be considered by industry during selection of almond.

The nervous and visual systems of onychophorans and tardigrades: learning about arthropod evolution from their closest relatives.

Understanding the origin and evolution of arthropods requires examining their closest outgroups, the tardigrades (water bears) and onychophorans (velvet worms). Despite the rise of molecular techniques, the phylogenetic positions of tardigrades and onychophorans in the panarthropod tree (onychophorans + tardigrades + arthropods) remain unresolved. Hence, these methods alone are currently insufficient for clarifying the panarthropod topology. Therefore, the evolution of different morphological traits, such as one of the most intriguing features of panarthropods-their nervous system-becomes essential for shedding light on the origin and evolution of arthropods and their relatives within the Panarthropoda. In this review, we summarise current knowledge of the evolution of panarthropod nervous and visual systems. In particular, we focus on the evolution of segmental ganglia, the segmental identity of brain regions, and the visual system from morphological and developmental perspectives. In so doing, we address some of the many controversies surrounding these topics, such as the homology of the onychophoran eyes to those of arthropods as well as the segmentation of the tardigrade brain. Finally, we attempt to reconstruct the most likely state of these systems in the last common ancestors of arthropods and panarthropods based on what is currently known about tardigrades and onychophorans.

Almond By-Products Substrates as Sustainable Amendments for Green Bean Cultivation.

Almond processing generates a high quantity of by-products, presenting the untapped potential for alternative applications and improved sustainability in production. This study aimed to evaluate whether the incorporation of almond by-products (hulls/shells) can improve the biochemical characteristics of green bean pods when used as an alternative to traditional growing media in green bean plants. Four substrates were prepared: the Control substrate (C): 70% peat + 30% perlite; substrate (AS): 70% peat + 30% shells; substrate (AH): 70% peat + 30% perlite + 1 cm hulls as mulch; substrate (MIX): 70% peat + 15% shells + 15% hulls. Plants were grown in each of these substrates and subjected to two irrigation levels, 100% and 50% of their water-holding capacity. Biochemical parameters (photosynthetic pigments, total phenolics, flavonoids, ortho-diphenols, soluble proteins, antioxidant capacity) and color were evaluated in the harvested pods. Results showed that pods from plants growing in AH substrate presented statistically significant higher values in their total phenolic content, while AS and MIX substrates did not reveal significant benefits. Summarily, this study highlights the potential of almond hulls as a promising medium for green bean cultivation, particularly when employed as mulch. Further research is recommended to gain a more comprehensive understanding of the application of almond by-products as natural fertilizers/mulch.

Impacts of Climate Change and Mitigation Strategies for Some Abiotic and Biotic Constraints Influencing Fruit Growth and Quality.

Factors such as extreme temperatures, light radiation, and nutritional condition influence the physiological, biochemical, and molecular processes associated with fruit development and its quality. Besides abiotic stresses, biotic constraints can also affect fruit growth and quality. Moreover, there can be interactions between stressful conditions. However, it is challenging to predict and generalize the risks of climate change scenarios on seasonal patterns of growth, development, yield, and quality of fruit species because their responses are often highly complex and involve changes at multiple levels. Advancements in genetic editing technologies hold great potential for the agricultural sector, particularly in enhancing fruit crop traits. These improvements can be tailored to meet consumer preferences, which is crucial for commercial success. Canopy management and innovative training systems are also key factors that contribute to maximizing yield efficiency and improving fruit quality, which are essential for the competitiveness of orchards. Moreover, the creation of habitats that support pollinators is a critical aspect of sustainable agriculture, as they play a significant role in the production of many crops, including fruits. Incorporating these strategies allows fruit growers to adapt to changing climate conditions, which is increasingly important for the stability of food production. By investing in these areas, fruit growers can stay ahead of challenges and opportunities in the industry, ultimately leading to increased success and profitability. In this review, we aim to provide an updated overview of the current knowledge on this important topic. We also provide recommendations for future research.

The role of ventral and preventral organs as attachment sites for segmental limb muscles in Onychophora.

BACKGROUND: The so-called ventral organs are amongst the most enigmatic structures in Onychophora (velvet worms). They were described as segmental, ectodermal thickenings in the onychophoran embryo, but the same term has also been applied to mid-ventral, cuticular structures in adults, although the relationship between the embryonic and adult ventral organs is controversial. In the embryo, these structures have been regarded as anlagen of segmental ganglia, but recent studies suggest that they are not associated with neural development. Hence, their function remains obscure. Moreover, their relationship to the anteriorly located preventral organs, described from several onychophoran species, is also unclear. To clarify these issues, we studied the anatomy and development of the ventral and preventral organs in several species of Onychophora. RESULTS: Our anatomical data, based on histology, and light, confocal and scanning electron microscopy in five species of Peripatidae and three species of Peripatopsidae, revealed that the ventral and preventral organs are present in all species studied. These structures are covered externally with cuticle that forms an internal, longitudinal, apodeme-like ridge. Moreover, phalloidin-rhodamine labelling for f-actin revealed that the anterior and posterior limb depressor muscles in each trunk and the slime papilla segment attach to the preventral and ventral organs, respectively. During embryonic development, the ventral and preventral organs arise as large segmental, paired ectodermal thickenings that decrease in size and are subdivided into the smaller, anterior anlagen of the preventral organs and the larger, posterior anlagen of the ventral organs, both of which persist as paired, medially-fused structures in adults. Our expression data of the genes Delta and Notch from embryos of Euperipatoides rowelli revealed that these genes are expressed in two, paired domains in each body segment, corresponding in number, position and size with the anlagen of the ventral and preventral organs. CONCLUSIONS: Our findings suggest that the ventral and preventral organs are a common feature of onychophorans that serve as attachment sites for segmental limb depressor muscles. The origin of these structures can be traced back in the embryo as latero-ventral segmental, ectodermal thickenings, previously suggested to be associated with the development of the nervous system.

An updated world checklist of velvet worms (Onychophora) with notes on nomenclature and status of names.

More than a decade has passed since the publication of the only world checklist available for Onychophora. During this period, numerous nomenclatural acts and taxonomic changes have been suggested within the group and a wealth of novel data has been published on many taxa. Herein, the up-to-date taxonomic scenario within Onychophora is presented, with appraisal of name status. This checklist covers both extant (Peripatidae and Peripatopsidae) and fossil taxa, and each species is accompanied by information on synonyms, type designation, holotype location, type locality, and language of original description. Additional remarks include nomenclatural inconsistencies, synonymizations, name misspellings, conflicting collecting event data, availability of taxonomically informative molecular data, etc. According to the data, 237 species are currently assigned to Onychophora: 140 of Peripatopsidae, 92 of Peripatidae, and five fossil species with unclear relationship to extant taxa. Since the previous checklist, 37 species have been added to Onychophora, representing an increase of 18.5% in the diversity described for the group. Yet, taxonomic descriptions seem slow-paced, with an average of 3.6 onychophoran species being described annually. From the taxonomic standpoint, 216 species are valid, although many of them require morphological revision and molecular characterization; 21 species exhibit major taxonomic ambiguities and have been regarded as nomina dubia. Recurrent taxonomic issues identified in the literature include inaccurate collecting event data, doubtful taxonomic assignment of molecular sequences, and non-observance of nomenclatural rules. These and other taxonomic aspects are addressed herein in the light of the directives established by the International Code of Zoological Nomenclature.

Degradation of Agro-Industrial Wastewater Model Compound by UV-A-Fenton Process: Batch vs. Continuous Mode.

The degradation of a model agro-industrial wastewater phenolic compound (caffeic acid, CA) by a UV-A-Fenton system was investigated in this work. Experiments were carried out in order to compare batch and continuous mode. Initially, batch experiments showed that UV-A-Fenton at pH 3.0 (pH of CA solution) achieved a higher generation of HO•, leading to high CA degradation (>99.5%). The influence of different operational conditions, such as H2O2 and Fe2+ concentrations, were evaluated. The results fit a pseudo first-order (PFO) kinetic model, and a high kinetic rate of CA removal was observed, with a [CA] = 5.5 × 10−4 mol/L, [H2O2] = 2.2 × 10−3 mol/L and [Fe2+] = 1.1 × 10−4 mol/L (kCA = 0.694 min−1), with an electric energy per order (EEO) of 7.23 kWh m−3 order−1. Under the same operational conditions, experiments in continuous mode were performed under different flow rates. The results showed that CA achieved a steady state with higher space-times (θ = 0.04) in comparison to dissolved organic carbon (DOC) removal (θ = 0−0.020). The results showed that by increasing the flow rate (F) from 1 to 4 mL min−1, the CA and DOC removal rate increased significantly (kCA = 0.468 min−1; kDOC = 0.00896 min−1). It is concluded that continuous modes are advantageous systems that can be adapted to wastewater treatment plants for the treatment of real agro-industrial wastewaters.

Treatment of Winery Wastewater by Combined Almond Skin Coagulant and Sulfate Radicals: Assessment of HSO5- Activators.

The large production of wine and almonds leads to the generation of sub-products, such as winery wastewater (WW) and almond skin. WW is characterized by its high content of recalcitrant organic matter (biodegradability index < 0.30). Therefore, the aim of this work was to (1) apply the coagulation-flocculation-decantation (CFD) process with an organic coagulant based on almond skin extract (ASE), (2) treat the organic recalcitrant matter through sulfate radical advanced oxidation processes (SR-AOPs) and (3) evaluate the efficiency of combined CFD with UV-A, UV-C and ultrasound (US) reactors. The CFD process was applied with variation in the ASE concentration vs. pH, with results showing a chemical oxygen demand (COD) removal of 61.2% (0.5 g/L ASE, pH = 3.0). After CFD, the germination index (GI) of cucumber and corn seeds was ≥80%; thus, the sludge can be recycled as fertilizer. The SR-AOP initial conditions were achieved by the application of a Box-Behnken response surface methodology, which described the relationship between three independent variables (peroxymonosulfate (PMS) concentration, cobalt (Co2+) concentration and UV-A radiation intensity). Afterwards, the SR-AOPs were optimized by varying the pH, temperature, catalyst type and reagent addition manner. With the application of CFD as a pre-treatment followed by SR-AOP under optimal conditions (pH = 6.0, [PMS] = 5.88 mM, [Co2+] = 5 mM, T = 343 K, reaction time 240 min), the COD removal increased to 85.9, 82.6 and 80.2%, respectively, for UV-A, UV-C and US reactors. All treated wastewater met the Portuguese legislation for discharge in a municipal sewage network (COD ≤ 1000 mg O2/L). As a final remark, the combination of CFD with SR-AOPs is a sustainable, safe and clean strategy for WW treatment and subproduct valorization.

Composition of Nuts and Their Potential Health Benefits-An Overview.

The possibility that nut intake may defend human health is an interesting point of view and has been investigated worldwide. Consequently, nuts are commonly promoted as healthy. In recent decades, the number of investigations proposing a correlation between nut consumption and a decrease in the risk of key chronic diseases has continued to increase. Nuts are a source of intake of fiber, and dietary fiber is associated with a reduced occurrence of obesity and cardiovascular diseases. Nuts likewise provide minerals and vitamins to the diet and supply phytochemicals that function as antioxidant, anti-inflammatory, and phytoestrogens agents and other protective mechanisms. Therefore, the main goal of this overview is to summarize current information and to describe the utmost new investigation concerning the health benefits of certain nuts.

Genome size and chromosome number in velvet worms (Onychophora).

The Onychophora (velvet worms) represents a small group of invertebrates (~180 valid species), which is commonly united with Tardigrada and Arthropoda in a clade called Panarthropoda. As with the majority of invertebrate taxa, genome size data are very limited for the Onychophora, with only one previously published estimate. Here we use both flow cytometry and Feulgen image analysis densitometry to provide genome size estimates for seven species of velvet worms from both major subgroups, Peripatidae and Peripatopsidae, along with karyotype data for each species. Genome sizes in these species range from roughly 5-19 pg, with densitometric estimates being slightly larger than those obtained by flow cytometry for all species. Chromosome numbers range from 2n = 8 to 2n = 54. No relationship is evident between genome size, chromosome number, or reproductive mode. Various avenues for future genomic research are presented based on these results.

Fungal diversity associated to the olive moth, Prays Oleae bernard: a survey for potential entomopathogenic fungi.

Olive production is one of the main agricultural activities in Portugal. In the region of Trás-os-Montes, this crop has been considerably affected by Prays oleae. In order to evaluate the diversity of fungi on Prays oleae population of Trás-os-Montes olive orchards, larvae and pupae of the three annual generations (phyllophagous, antophagous and carpophagous) were collected and evaluated for fungal growth on their surface. From the 3,828 larvae and pupae, a high percentage of individuals exhibited growth of a fungal agent (40.6%), particularly those from the phyllophagous generation. From all the moth generations, a total of 43 species from 24 genera were identified, but the diversity and abundance of fungal species differed between the three generations. Higher diversity was found in the carpophagous generation, followed by the antophagous and phyllophagous generations. The presence of fungi displaying entomopathogenic features was highest in the phyllophagous larvae and pupae, with Beauveria bassiana as the most abundant taxa. The first report of Beauveria bassiana presence on Prays oleae could open new strategies for the biocontrol of this major pest in olive groves since the use of an already adapted species increases the guarantee of success of a biocontrol approach. The identification of antagonistic fungi able to control agents that cause major olive diseases, such as Verticillium dahliae, will benefit future biological control approaches for limiting this increasingly spreading pathogen.

Tolerance and bioaccumulation of copper by the entomopathogen Beauveria bassiana (Bals.-Criv.) Vuill. exposed to various copper-based fungicides.

This work evaluates for the first time the relationships between copper-tolerance, -solubilization and -bioaccumulation in the entomopathogen Beauveria bassiana exposed to Bordeaux mixture, copper oxychloride or copper hydroxide. Bordeaux mixture was highly detrimental to fungus, by inhibiting the growth totally at the recommended dose (RD) and 2×RD. Copper hydroxide and copper oxychloride were found to be less toxic, reducing fungus growth, sporulation and conidial germination in an average of 29  %, 30 % and 58 %, respectively. These two copper forms were the easiest to solubilize, to precipitate and the most accumulated by B. bassiana, suggesting the involvement of all these processes on fungus copper-tolerance.

Preliminary Evaluation of the Application of Algae-Based Biostimulants on Almond.

To improve almond performance under water limitations, the use of algae-based biostimulants may become a useful tool to reduce drought stress. However, besides possible effects on plant behavior, changes in fruit characteristics must also be considered. In this work, a preliminary study on the effect of two levels of an Ascophyllum nodosum-based biostimulant in the chemical characteristics of fruits from rain-fed cv. Marinada almond trees was carried out. The use of the recommended manufacturer's dosage resulted in a decreased content of soluble sugars and proteins when compared to the use of half the recommended dosage and the control assays (water only). Similarly, and although no significant differences were recorded, the content of bioactive compounds (ortho-diphenols, total phenolics, and flavonoids) tended to increase in non-treated trees. Finally, sensory analysis of treated and non-treated fruits did not record any changes in the perceived attributes, showing that no negative effects on consumers' acceptance will be caused by the application of this product. Long-term studies are needed to further confirm these results, also aimed at the monitoring of leaf gas exchange and water status parameters of trees.

Red Fruits Composition and Their Health Benefits-A Review.

The probability that fruit ingestion may protect human health is an intriguing vision and has been studied around the world. Therefore, fruits are universally promoted as healthy. Over the past few decades, the number of studies proposing a relationship between fruit intake and reduced risk of major chronic diseases has continued to grow. Fruits supply dietary fiber, and fiber intake is linked to a lower incidence of cardiovascular disease and obesity. Fruits also supply vitamins and minerals to the diet and are sources of phytochemicals that function as phytoestrogens, antioxidant and anti-inflammatory agents, and other protective mechanisms. So, this review aims to summarize recent knowledge and describe the most recent research regarding the health benefits of some selected red fruits.