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The zebrafish is a valuable model organism that is widely used in studies of vertebrate development. In the laboratory, zebrafish embryonic development is normally carried out at 28.5°C. In this study, we sought to determine whether it was possible to modify the speed of embryonic development through the use of short- and long-term variations in incubation temperature. After incubation at 20°C-32°C, most early-stage embryos survived to the epiboly stage, whereas more than half of the embryos died at20°C or32°C. The rate of development differed between embryos incubated at the lowest (18°C) and highest (34°C) temperatures: a difference of 60 min was observed at the 2-cell stage and 290 min at the 1k-cell stage. When blastulae that had developed at 28°C were transferred to a temperature lower than 18°C for one or more hours, they developed normally after being returned to the original 28°C. Analyses using green fluorescent protein

Burst swimming velocity (Uburst) was compared between wild and hatchery-reared chum salmon fry. In the hatchery-reared fry, Uburst was significantly correlated with the condition factor, but not with the body mass and fork length. In the wild-reared fry, on the other hand, Uburst ranged widely and did not correlate with condition factor. These suggest that well-balanced growth under satisfactory nutrient condition at the early developmental stage improves Uburst, and in the wild-reared fry, their cautiousness and various experiences may override the condition factor dependency of Uburst.

Marine aquaculture activities in the Sangihe Islands, more specifically in the waters of Talengen Bay, are not yet optimal and still utilize seeds from catches in nature. This study aims to determine the effect of using the Oodev hormone and obtain the best dose to stimulate the gonadal maturity level (TKG) of red snapper (Lutjanus sp.). The red snapper used comes from the catch in the waters of Talengen Bay and is also domesticated in floating net cages. The sampled fish used were 60 individuals with average body weight (30.5 g). The Oodev hormone dose treatment, namely; 0 mL/kg, 0.5 mL/kg, 1 mL/kg and 1.5 mL/kg. Oodev hormone was applied to commercial feed specifically for marine fish with the trademark Megami GR-3 orally for 30 days of rearing. The results showed that the administration of the Oodev hormone had a positive effect on the gonadal maturity level of red snapper, with the development of different levels of gonadal maturity in each treatment. The 0.5 mL/kg dose treatment resulted in the highest gonadal maturity level (TKG III). The maturation stage was followed by 1 mL/kg and 1.5 mL/kg (TKG II) treatments. Meanwhile, compared with the 0 mL/kg treatment, the developmental stage was still TKG I (immature). These findings provide Oodev hormone with 0.5 mL/kg dose could stimulate the red snapper gonad maturity level to mature for 30 days of rearing.

The microbiome in gastrointestinal tracts play an important role in regulating nutrient utilization and absorption, gut immune function, and host growth or development. This study was conducted to investigate the composition and dynamic distribution of caecal microbiota in pullets during the first 16 weeks. Growth performance, immune organs index, and intestinal morphology of pullets were analyzed at 3, 6, 12 and 16 weeks of age. The caecal contents were collected for microbiota analysis by 16S rRNA gene sequencing method. With advancing ages in pullets, the gradually increased average daily feed intake (ADFI), feed conversion ratio (FCR) and intestinal villus height, but the gradually decreased organs index of thymus and bursa were determined. Meanwhile, more abundant caecal bacterial communities were determined from pullets at 12 and 16 weeks of age than those at 3 and 6 weeks of age. Furthermore, the dominant microflora of pullets from different weeks of age were analyzed by using LEfSe: The higher abundance of Blautia, Prevotella, Alistipes, and Eggerthella were found at 6 weeks; Anaerostipes, Oscillospira, Enterococcus and Methanobrevibacter were determined at 12 weeks; and the higher abundance of Parabacteroides, Anaerofustis, Lactobacillus and Butyricimonas were determined at 16 weeks. Further functional predicted analysis by PICRUSt revealed that the endocrine system and carbohydrate metabolism were significantly developed at 3 weeks. The development of the immune system was predicted to be mainly during 6 to 12 weeks, while cardiovascular diseases and circulatory system were during 12 to 16 weeks. In addition, the significantly negative correlation between Bacteroides and villus height, the significantly negative correlation between growth parameters (ADFI and FCR) and Bacteroides, Oscillospira and Alistipes; and the significantly positive relations between growth parameters (ADFI and FCR) and Bilophila, Lactobacillus, Rikenella and Anaerofustis were determined by using Pearson analyses. In conclusion, our data demonstrated that growth performance and intestinal morphology correlate well with caecal microbiota, which could provide new insights to establish or develop nutritional strategies to manage the intestinal health or development of laying pullets.

Megaselia spiracularis Schmitz (Diptera: Phoridae) is a scuttle fly species of forensic importance and has been recorded colonizing human corpses in Malaysia, Korea and China. The necrophagous activity of M. spiracularis larvae on human corpses can be referred to estimate minimum postmortem interval (PMImin) in death investigation. However, developmental data of M. spiracularis derived from ambient fluctuating temperature is scarce and taxonomic descriptions to larval instars for species identification are still lacking. This research was conducted to address these limitations by studying the development of M. spiracularis at fluctuating ambient temperatures. From each sampling occasion, larval body lengths were obtained and samples were mounted on slides to determine larval instars based on taxonomic characteristics of the cephalopharyngeal skeleton, anterior spiracles and posterior spiracles. At fluctuating temperatures of 25–37 °C and relative humidity of 47–93%, developmental duration of egg stage was 7–9.5 h; first instar larva, 9.5–17 h; second instar, 6–12 h, feeding third instar, 18–36 h; post-feeding third instar, 6–12 h; and pupa, 168–204 h. Accumulated degree hours for each developmental stage based on fluctuating ambient temperatures were also determined. The size of M. spiracularis larvae varied from 0.64–0.78 mm after eclosion to 5.20–5.95 mm before entering post feeding phase. The first, second and third instar larva could be distinguished mainly by the morphology of cephalopharyngeal skeletons, anterior spiracles and posterior spiracles. Developmental durations of M. spiracularis larvae and the larval instars can be used as references to estimate PMImin as this species is likely to be featured in future forensic cases.

Recent discovery of a previously unknown outcrop in the vicinity of the Zengővárkony lime-kilns (Mecsek Mountains, Hungary) provided a few identifiable upper Oxfordian brachiopods that exhibit a truly Mediterranean (Tethyan) character. Dating of the outcrop is based on a rich ammonite fauna: Benetticeras benettii; Trimarginites ex gr. trimarginatus; Orthosphinctes (Orthosphinctes) ex gr. tiziani clearly indicate the Late Oxfordian. The brachiopod fauna indicates a deep-water marine environment and well-oxygenated sea floor. Nucleata bouei and Pygope catulloi are recorded for the first time from the Mecsek Mountains. A pathologic specimen of Pygope catulloi is also recognized. Its ventral valve was injured in an early developmental stage that caused deformation of the left side, which was overgrown by the healthy right side and created an asymmetric adult shell shape. Cause of the injury is unclear but it provides further evidence for subsequent healing of brachiopods after being injured. This is the first description and illustration of Oxfordian brachiopods from the Mecsek Mountains, Hungary. The occurrence of Tethyan originating pygopid brachiopods in the Oxfordian strengthens earlier observations that from the Bathonian/Callovian Tethyan influence became overwhelming in the Mecsek Mountains fauna. Pygope catulloi strengthens records from Algeria that pygopid brachiopods may have occurred very early on the periphery of the Western Tethys.

Sticky layer of Clarias gariepinus eggs cause the clumping of eggs immediately after fertilization, this create low oxygen, and increase pathogenic infection, thereby reducing fertilization and hatching rate hence reduces in mass production of C. gariepinus seed. Although various substances are use in the removal of the sticky layer, no information is available about the use of watermelon. Therefore, the present study, aim in exploring the potential of using watermelon in removing the sticky layer of C. gariepinus to enhance production. Fertilized eggs were treated with watermelon juice (T1) for comparison with 14 g L-1 milk solution (T2 as de-adhesion reference) and water as control (T0 as adhesion reference). The treatments were agitated with aeration for 30 minutes, rinsed with water, and poured on respective netted hatching trough, with water recirculation for incubation at 27 0C for 24 h period. Except for T0, no clumping of eggs was observed in the remaining groups. Slight variation in attaining a developmental stage among eggs was observed in eggs treated with watermelon. Hatching and survival rate in T2 are higher than T1 and T0 (P0.05), more should be explored before adoption of watermelon for de-adhesion and incubation of C. gariepinus eggs.

Seedling emergence, survival, morphological and physiological traits, and oxidative stress resistance of southwestern white pine (Pinus strobiformis Engelm.) were studied in response to warming treatments applied during embryogenesis, germination, and early seedling growth. Daytime air temperature surrounding cones in tree canopies was warmed by +2.1°C during embryo development. Resulting seeds and seedlings were assigned to three thermal regimes in growth chambers, with each regime separated by 4°C to encompass the wide range of temperatures observed over space and time across the species’ range, plus the effect of heat waves coupled with a high carbon emissions scenario of climate warming. The embryo warming treatment reduced percent seedling emergence in all germination and growth environments and reduced mortality of seedlings grown in the warmest environment. Warm thermal regimes during early seedling growth increased subsequent seedling resistance to oxidative stress and transpirational water use. Experimental warming during seed development, germination, and seedling growth affected seedling emergence and survival. Oxidative stress resistance, morphology, and water relations were affected only by warming imposed during germination and seedling growth. This work explores potential outcomes of climate warming on multiple dimensions of seedling performance and uniquely illustrates that plant responses to heat vary with plant developmental stage in addition to the magnitude of temperature change.

Phytophthora diseases are serious and often fatal in arecanut causing huge losses to farmers (50–100%) if timely and proper management measures aren’t adopted. Among Phytophthora diseases, fruit rot disease predominantly exists in all the arecanut growing regions receiving heavy rainfall during the south-west monsoon season. The extended manifestation of pathogen causes crown and bud rot disease which prevails till November–December due to prolonged rains and congenial weather factors. The role of Phytophthora in causing these diseases of arecanut was established in the early 1900’s and later the Phytophthora meadii was identified as a causal agent. Though the diseases are sporadic, their management in the field situation is a challenging task due to intense rainfall and the variability in the pathogen. The continuous heavy rainfall during the tender arecanut developmental stage, non-availability of professional climbers coupled with the absence of machinery for powerful spraying, occurrence of aggressive pathotypes and lack of Phytophthora resistant/tolerant arecanut varieties are the major hindrance for taking up effective disease management measures. The present review has attempted to compile the earlier findings, present status of this century-old disease, and discusses future strategies for the development of effective remedies for this serious malady of the areca palm.

This article highlights the second stage of athletic development, the Developmental Stage. In this article we build awareness in growth and maturation, highlighting a 3-stage process through the use of the Khamis-Roche protocol. We covered an introduction to Strength Development for athletes in the developmental stage. Speed Development was addressed in a progressive way to teach critical positions and we introduced ways to assess a developing athlete from ages 12–15.

The large Japanese field mouse (Apodemus speciosus) is a small rodent species endemic to Japan. The genetic characteristics of A. speciosus include different chromosome numbers within the same species. Furthermore, A. speciosus has been used in radiation and genetic research. In the present study, a pregnant A. speciosus was obtained, and histochemical analysis of the implanted embryos was performed and compared with the developmental stages of the mouse (Mus musculus). Although there were some differences, the structures of the implanted embryos, including the primitive streak and placenta of A. speciosus were similar to those of mouse. Our study will be important for the construction of a developmental atlas of A. speciosus.

Most of the yeast bypasses the developmental stage from simple unicellular yeast to elongated structure like hyphae. Regulation of this transition is governed by various quorum sensing and signalling molecules produced under different conditions of growth, that differ significantly, both physiologically and chemically. The evidence of fungal quorum sensing was uncovered ten years ago after the discovery of farnesol as first eukaryotic quorum sensing molecules in Candida albicans. In addition to farnesol, tyrosol was identified as second quorum sensing molecules in C. albicans controlling physiological activities. After the discovery of farnesol and tyrosol, regulation of morphogenesis through the production of chemical signalling molecules such as isoamyl alcohol, 2-phenylethyl alcohol, 1-dodecanol, E-nerolidol, etc. is reported in C. albicans. Some of the evidence suggests that the budding yeast Saccharomyces cerevisiae exhibits this type of regulation and the signals are regulated by aromatic alcohols which are the end product of amino acid metabolism. The effects of these molecules on morphogenesis are not similar in both yeasts, making comparisons hard. It is hypothesized that these signals works in microorganisms to derive a competitive advantage. Here, we present an example for utilization of competitive strategy by C. albicans and S. cerevisiae over other microorganisms.

Schistosomiasis is a chronic and painful disease of poverty caused by the flatworm parasite Schistosoma. Drug discovery for antischistosomal compounds predominantly employs in vitro whole organism (phenotypic) screens against two developmental stages of Schistosoma mansoni, post-infective larvae (somules) and adults. We generated two rule books and associated scoring systems to normalize 3898 phenotypic data points to enable machine learning. The data were used to generate eight Bayesian machine learning models with the Assay Central software according to parasite's developmental stage and experimental time point (≤24, 48, 72, and >72 h). The models helped predict 56 active and nonactive compounds from commercial compound libraries for testing. When these were screened against S. mansoni in vitro, the prediction accuracy for active and inactives was 61% and 56% for somules and adults, respectively; also, hit rates were 48% and 34%, respectively, far exceeding the typical 1-2% hit rate for traditional high throughput screens.

Accurate characterization of oocyte development is essential to understanding foundational aspects of reproductive biology and successful management of Pacific halibut (Hippoglossus stenolepis). Here this study provides complete histological descriptions for eight oocyte developmental stages in addition to postovulatory follicles and demonstrates the potential for oocyte size frequency distribution to act as a proxy for ovarian developmental stage and future maturity assessments. Importantly, it provides the first histological evidence that Pacific halibut have a group‐synchronous ovarian developmental pattern with determinate fecundity and support for their batch‐spawning strategy.

BACKGROUND The vertebrate jaw is thought to have evolved through developmental modification of the mandibular arch. An extant jawless vertebrate, the lamprey, possesses a structure called "velum"-a mandibular arch derivative-in addition to the oral apparatus. This leads us to assess the velum's possible contribution to the evolution of jaws. RESULTS The velar muscles develop from progenitor cells distinct from those from which the oral muscles develop. In addition, the oral and velar regions originate from the different sub-population of the trigeminal neural crest cells (NCCs): the former region receives NCCs from the midbrain, whereas the latter region receives NCCs from the anterior hindbrain. The expression of patterning genes (eg, DlxA and MsxA) is activated at a later developmental stage in the velum compared to the oral region, and more importantly, in different cells from those in the oral region. CONCLUSION The lamprey mandibular arch consists of two developmental units: the anterior oral unit and the posterior velar unit. Because structural elements of the lamprey velum may be homologous to the jaw, the evolution of vertebrate jaws may have occurred by the velum being released from its functional roles in feeding or respiration in jawless vertebrates.

Parasitic castration of bivalves by trematodes is common, and may significantly reduce the reproductive capacity of ecologically important species. Understanding the intensity of infection is desirable, as it can indicate the time that has passed since infection, and influence the host's physiological and reproductive response. In addition, it is useful to know the developmental stage of the trematode, to understand trematode population trends and reproductive success. However, most existing methods (e.g. visually estimating the degree of infection) to assess intensity are approximate only and not reproducible. Here, we present a method to accurately quantify the percentage of bivalve gonad filled with digenean trematode tissue, based on small squashes of gonad tissue rapidly photographed under light microscopy. A maximum of 15 photographs is required to determine the percentage of the whole gonad occupied by trematodes with a minimum of 90% confidence, with smaller mussels requiring fewer. In addition, the stage of trematode infection can be assessed because full sporocysts, spent sporocysts and free cercariae are clearly distinguishable. Although variation exists in the distribution of trematodes in gonad tissue, and thus in the estimate of percentage of the gonad filled with trematodes, this method represents a marked improvement on current coarse assessments of infection which typically focus on binary presence/absence measures. This technique can be used to facilitate a more sophisticated understanding of host–parasite interactions in bivalves, and can inform the conservation and reproductive biology of environmentally crucial species.

Variability in the contents of representative secondary metabolites in Gentiana pneumonanthe individuals was investigated in relation to population origin, plant parts and developmental stage. Chem...

Enterovirus A71 (EV71) is one of the major causative agents of hand, foot and mouth disease (HFMD), and is sometimes associated with severe central nervous system syndromes. Vaccines against EV71 infection have been developed or are in development in several countries and few have been licensed in China. In response to requests from some of these countries, WHO convened a working group meeting in Shanghai China from 11 to 12 September 2019 to develop WHO Recommendations to assure the quality, safety and efficacy of EV71 vaccines. Meeting participants included members of the drafting group, experts from vaccine developers, manufacturers, regulators and academia. The epidemiology of EV71, as well as the development, regulation and standardization of EV71 vaccines were reviewed in the meeting. Information on R&D, manufacturing, quality control and standardization of EV71 vaccines was presented by vaccine developers, manufacturers and regulators. Based on their experience, the working group discussed the main principles that would determine WHO’s position on quality, safety and efficacy of EV71 vaccines. The working group agreed to develop WHO Recommendations to assure the quality, safety and efficacy of inactivated EV71 vaccines with a scope covering only whole virus inactivated vaccines. Other type of vaccines, such as EV71 virus-like particles (VLPs) will not be covered as they are still at the developmental stage. The outline of the document was agreed and will follow the usual style of WHO recommendations. It was also agreed to submit the draft Recommendations for review and adoption to the WHO ECBS in 2020 following discussion at a WHO informal consultation, which will include NRAs and vaccine manufacturers.

This experiment was conducted to identify the morphological characteristics of elevated atmospheric CO2 concentration of Kalanchoe blossfeldiana cultivars under different daylength treatments, in order to apply the carbon reduction method. CO2 treatment took place between 22:00 and 04:00 with an average of 400 μmol·mol-1 (control) and 800 μmol·mol-1 on Kalanchoe blossfeldiana ‘Lion’ and ‘Fikalrudak’. The photoperiod was controlled using an 8 h (08:00 to 16:00), 12 h (06:00 - 18:00), and 16 h (04:00 - 20:00) light-dark cycle. The number of leaves increased more in plants under the 800 μmol·mol-1 treatment than those in the control, with a photoperiod of 8 h and 12 h in both K. ‘Lion’ and 8 h in K. ‘Fikalrudak’. Plant height was lower in plants treated with 800 μmol·mol-1 CO2 than under the control treatment with an 8-h photoperiod of 8 h, regardless of K. blossfeldiana cultivars. In K. ‘Lion’, immature leaf length and width increased more in plants treated with 800 μmol·mol-1 CO2 than in the control, with a photoperiod of 8 h and 12 h. However, the leaf length and width of K. ‘Fikalrudak’ was not affected by CO2 concentration. The nodes were produced primarily by increasing CO2 concentration to 800 μmol·mol-1, regardless of daylength and K. blossfeldiana cultivars. Flowering time was not affected by daylength, however, K. ‘Lion’ bloomed within the photoperiod of 8 h and 12 h and K. ‘Fikalrudak’ bloomed only under a photoperiod of 8 h. K. blossfeldiana cultivars bloomed effectively under less light availability, regardless of CO2 concentration. These results showed that a CO2-rich enrichment promotes leaf and node production of Kalanchoe blossfeldiana cultivars under short-day conditions. Since high concentration of CO2 enrichment can have an adverse effect on flowering quality, appropriate CO2 enrichment methods for the developmental stage and cultivars should be applied on a case-by-case basis.

Summary Introduction: Bastard balm grows in forests, in central and southeastern part of Europe. The herb of this species is rich in phenolics, mainly flavonoids, phenolic acids and coumarins. The plant is used in traditional European medicine, in digestive problems and for aromatizing tobacco and alcohol products. Objective: The purpose of this study was to determine the influence of shading on bastard balm development and the accumulation of phenolics in its herb, with special respect to coumarin as a quality marker of this raw material. Methods: The plants were cultivated in full sunlight, in 30% and 50% shade provided by shading nets. The herb was harvested from plants in the third year of vegetation, at four subsequent developmental stages and then subjected to chemical evaluation. In the raw material, the total contents of flavonoids, phenolic acids and coumarins was determined. The content of coumarin was analyzed using HPLC-DAD. Results: Plants grown in 30% shade produced the highest number of flowers and seeds. They produced the highest mass of herb at the beginning of the seed-setting stage. The plants grown in full sunlight revealed the highest content of flavonoids and phenolic acids, especially during flowering and at the beginning of the seed-setting stage. The mass of herb obtained by plants cultivated at deep (50%) shade was the lowest, however, the content of coumarin in these plants was the highest. Conclusion: The influence of shade on bastard balm was expressed by the herb mass and coumarin content increment. The plants thrived best in 30% shade, both in terms of flowering abundance and the mass of herb, whereas those from 50% shade were the richest in coumarin.

Volatile organic compounds (VOC) are characteristic for different fungal species. However, little is known about VOC changes during development and their biological role. Therefore, we established a laboratory cultivation system in modified crystallizing dishes for analyzing VOC during fruiting body development of the dikaryotic strain Cyclocybe aegerita AAE-3 as well as four monokaryotic offspring siblings exhibiting different fruiting phenotypes. From these, VOC were extracted directly from the headspace (HS) and analyzed by means of gas chromatography-mass spectrometry (GC-MS). For all tested strains, alcohols and ketones, including oct-1-en-3-ol, 2-methylbutan-1-ol and cyclopentanone, were the dominant substances in the HS of early developmental stages. In the dikaryon, the composition of the VOC altered with ongoing fruiting body development and, even more drastically, during sporulation. At the latter stage, sesquiterpenes, especially Δ6-protoilludene, α-cubebene and δ-cadinene, were the dominant substances. After sporulation, the amount of sesquiterpenes decreased, while additional VOC, mainly octan-3-one, appeared. In the HS of the monokaryons, less VOC were present of which all were detectable in the HS of the dikaryon C. aegerita AAE-3. The results of the present study show that the volatilome of C. aegerita changes considerably depending on the developmental stage of the fruiting body.

Growing seedlings and transplants (plugs and liners) in plant factories with electric lighting enables control of environments to produce consistent and uniform plants. In addition, as light emitting diodes (LEDs) become common light sources for sole-source lighting in plant factories, the ability to manipulate the photon spectrum makes it possible to elicit desirable quality attributes of transplants. However, the effects of specific wavelengths on plant traits can depend on other wavelengths, light quantity, photoperiod, species, developmental stage, and other environmental parameters. In addition, there can be trade-offs between crop growth (yield) and quality attributes. Therefore, it is challenging to identify and optimize LED lighting for transplant and seedling production, and likely, there is not a “one-size-fits-all” photon spectrum. In this chapter, we discuss recent research findings on how the quality, quantity, and daily duration of sole-source LED lighting influence growth and development characteristics of floriculture and vegetable transplants produced from seed or stem cuttings, as well as microgreens.

The mechanisms supporting regeneration and successful recovery of function have fascinated scientists and the general public for quite some time, with the earliest description of regeneration occurring in the 8th century BC through the Greek mythological story of Prometheus. While most animals demonstrate the capacity for wound-healing, the ability to initiate a developmental process that leads to a partial or complete replacement of a lost structure varies widely among animal taxa. Variation also occurs within single species based on the nature and location of the wound and the developmental stage or age of the individual. Comparative studies of cellular and molecular changes that occur both during, and following, wound healing may point to conserved genomic pathways among animals of different regenerative capacity. Such insights could revolutionize studies within the field of regenerative medicine. In this review, we focus on several closely related species of Lumbriculus (Clitellata: Lumbriculidae), as we present a case for revisiting the use of an annelid model system for the study of regeneration. We hope that this review will provide a primer to Lumbriculus biology not only for regeneration researchers but also for STEM teachers and their students.

Endophytic bacteria play potentially important roles in the processes of plant adaptation to the environment. Understanding the composition and dynamics of endophytic bacterial communities under heavy metal (HM) stress can reveal their impacts on host development and stress tolerance. In this study, we investigated root endophytic bacterial communities of different rice cultivars grown in a cadmium (Cd)-contaminated paddy field. These rice cultivars are classified into low (RBQ, 728B, and NX1B) and high (BB and S95B) levels of Cd-accumulating capacity. Our metagenomic analysis targeting 16S rRNA gene sequence data reveals thatProteobacteria,Firmicutes,Actinobacteria,Acidobacteria,Bacteroidetes, andSpirochaetesare predominant root endophytic bacterial phyla of the five rice cultivars that we studied. Principal coordinate analysis shows that the developmental stage of rice governs a larger source of variation in the bacterial communities compared to that of any specific rice cultivar or of the root Cd content. Endophytic bacterial communities during the reproductive stage of rice form a more highly interconnected network and exhibit higher operational taxonomic unit numbers, diversities, and abundance than those during the vegetative stage. Forty-five genera are significantly correlated with Cd content in rice root, notably including positive-correlatingGeobacterandHaliangium; and negative-correlatingPseudomonasandStreptacidiphilus. Furthermore, Phylogenetic Investigation of Communities by Reconstruction of Unobserved States analysis shows that functional pathways, such as biosynthesis of siderophore and type II polyketide products, are significantly enhanced during the reproductive stage compared to those during the vegetative stage under Cd stress. The isolated endophytic bacteria from the Cd-contaminated rice roots display high Cd resistance and multiple traits that may promote plant growth, suggesting their potential application in alleviating HM stress on plants. This study describes in detail for the first time the assemblage of the bacterial endophytomes of rice roots under Cd stress and may provide insights into the interactions among endophytes, plants, and HM contamination.

Epigenome is susceptible to modulation by environmental pressures—namely, through alterations in global DNA methylation, impacting the organism condition and, ultimately, reverberating on the phenotype of the subsequent generations. Hence, an intergenerational study was conducted, aiming to clarify the influence of genotoxicants on global DNA methylation of the crayfish Procambarus clarkii. Two subsequent generations were exposed to the herbicide penoxsulam (Px, 23 µg·L−1) and to the genotoxicant model ethyl methanesulfonate (EMS, 5 mg·L−1). Px did not induce changes in DNA methylation of adult crayfish (F0). However, the hypomethylation occurring in unexposed F1 juveniles demonstrated that the history of exposure per se can modulate epigenome. In F1 descendants of the Px-exposed group, methylome (hypermethylated) was more affected in males than in females. EMS-induced hypomethylation in adult females (F0), also showed gender specificity. In addition, hypomethylation was also observed in the unexposed F1 crayfish, indicating an intergenerational epigenetic effect. The modulatory role of past exposure to penoxsulam or to EMS also showed a dependency on the crayfish developmental stage. Overall, this research revealed that indirect experiences (events occurring in a predecessor generation) can have an impact even greater than direct experiences (present events) on the epigenetic dynamics.

Foxtail millet (Setaria italica) is rich in nutrients and extremely beneficial to human health. We identified and comprehensively analyzed 89 MADS-box genes in the foxtail millet genome. According to the classification of MADS-box genes in Arabidopsis thaliana and rice, the SiMADS-box genes were divided into M-type (37) and MIKC-type (52). During evolution, the differentiation of MIKC-type MADS-box genes occurred before that of monocotyledons and dicotyledons. The SiMADS-box gene structure has undergone much differentiation, and the number of exons in the MIKC-type subfamily is much greater than that in the M-type subfamily. Analysis of gene duplication events revealed that MIKC-type MADS-box gene fragment duplication accounted for the vast majority of gene duplication events, and MIKC-type MADS-box genes played a major role in the amplification of SiMADS-box genes. Collinearity analysis showed highest collinearity between foxtail millet and maize MADS-box genes. Analysis of tissue-specific expression showed that SiMADS-box genes are highly expressed throughout the grain-filling process. Expression analysis of SiMADS-box genes under eight different abiotic stresses revealed many stress-tolerant genes, with induced expression of SiMADS33 and SiMADS78 under various stresses warranting further attention. Further, some SiMADS-box proteins may interact to cope with external stress. This study provides insights for MADS-box gene mining and molecular breeding of foxtail millet in the future.

Coleoptera are currently considered a fundamental tool to help solve criminal investigations, allowing forensic entomologists to estimate post-mortem intervals and obtain other ecology-related information. Thanatophilus rugosus (Linnaeus, 1758) is an important necrophagous beetle distributed through most of the Palaearctic region, where it is readily found on human bodies and animal carcasses. In this study, the new thermal summation models for all the developmental stages of Thanatophilus rugosus are provided. Beetles were reared at six different constant and ecologically relevant temperatures (12, 14, 16, 18, 20, and 22 °C), and their developmental times were measured. Thermal summation constants were calculated for each developmental stage (egg, three larval instars, post-feeding stage, and pupa).

The use of whole animal models in toxicological studies is essential for understanding the physiological responses caused by chemical exposures. However, such studies can face reproducibility challenges due to unaccounted experimental parameters that can have a marked influence on toxicological outcomes. Zebrafish embryos and larvae are a popular vertebrate animal model for studying cellular, tissue, and organ responses to toxicant exposures. Despite the popularity of this system, standardized protocols that control for the influence of various experimental parameters and culture conditions on the toxicological response in these animals have not been widely adopted, making it difficult to compare findings from different laboratories. Here, we describe a detailed approach for designing and optimizing protocols to assess the impact of chemical exposures on the development and survival of zebrafish embryos and larvae. We first describe our standard procedure to determine two key toxicological thresholds, the maximum tolerable concentration (MTC) and the lethal concentration (LC50 , defined as that in which 50% of larvae die), in response to an exposure that persists from early development through larval maturation. We then describe two protocols to systematically test how key experimental parameters, including genetic background, culture media, animal density, volume, plate material, and developmental stage in which the embryos are exposed, alter the MTC and LC50 . Finally, we provide a step-by-step guide to assess the interaction between two chemicals using this model. These protocols will guide the standardization of toxicological studies using zebrafish and maximize reproducibility. © 2021 Wiley Periodicals LLC. Basic Protocol 1: Zebrafish embryo collection and culture, and establishment of the MTC and LC50 Basic Protocol 2: Evaluation of the impact of culture conditions on toxicant responses of zebrafish embryo and larvae Basic Protocol 3: Identification of the developmental window of sensitivity to toxicant exposure Basic Protocol 4: Testing interaction between multiple toxicants.

As the demand for drought hardy tree seedlings rises alongside global temperatures, there is a need to optimize nursery drought preconditioning methods to improve field performance of planted seedlings. This perspective article advocates for a more holistic approach to drought preconditioning research that considers the moderating role of plant developmental stage on the effects of drought preconditioning. We identify discrepancies in past studies of root growth potential (RGP) responses to drought preconditioning and highlight studies that suggest such discrepancies may result from inconsistencies among studies in the timing of drought preconditioning implementation. We then illustrate our perspective by presenting original research from an aeroponic RGP trial of 1st-year western larch (Larix occidentalis Nutt.) seedlings exposed to three soil moisture contents for 6months. We evaluated whether drought preconditioning could be used to increase the ratio of root: foliar tissue mass or enhance seedling physiological vigor during a subsequent growth period. Drought preconditioning was found to increase the ratio of root: foliar tissue mass and enhance seedling physiological vigor. Specifically, soil moisture content related negatively with new root biomass, positively with new foliar biomass, and negatively with the length and number of new roots (p

Buds usually possess mechanical or chemical protection and may also have secretory structures. We discovered an intricate secretory system in Ouratea castaneifolia (Ochnaceae) related to the protection of buds and young leaves. We studied this system, focusing on the distribution, morphology, histochemistry, and ultrastructure of glands during sprouting. Samples of buds and leaves were processed following the usual procedures for light and electron microscopy. Overlapping bud scales protect dormant buds, and each young leaf is covered with a pair of stipules. Stipules and scales possess a resin gland, while the former also possess an extrafloral nectary. Despite their distinct secretions, these glands are similar and comprise secreting palisade epidermis. Young leaves also possess marginal colleters. All the studied glands shared some structural traits, including palisade secretory epidermis and the absence of stomata. Secretory activity is carried out by epidermal cells. Functionally, the activity of these glands is synchronous with the young and vulnerable stage of vegetative organs. This is the first report of colleters and resin glands for O. castaneifolia. We found evidence that these glands are correlated with protection against herbivores and/or abiotic agents during a developmental stage that precedes the establishment of mechanical defenses.

The use of repellents is a unique measure of personal protection, which can avoid tick attachment and thus reduce the risk of tick-borne infections. In the European Union, the efficacy of the repellents coming onto the market has to be evaluated according to the guidelines published by the European Chemical Agency before registration. The United States Environmental Protection Agency has a similar role. Despite obvious differences in morphology and behavior, both these guidelines allow the use of nymph or adult female ticks for laboratory testing. Here, we provide evidence that sensitivity to diethyltoluamide (deet) (P < 0.0001) of Ixodes ricinus nymphs within the in vitro trial was significantly higher than in adult females. In the experiment, we also observed that feral ticks were less sensitive to repellent than were laboratory-reared ticks (P < 0.01) and that mobility decreased when the trial was repeated (P < 0.05). This study showed that the results of efficacy tests could vary significantly even when the protocol was conducted in accordance with the recommended methods. To refine the results of efficacy tests, we recommend a reevaluation of the guidelines, with emphasis on the developmental stage and origin of ticks.

Embedding and immobilisation of living cells and microorganisms is used in a variety of research and commercial applications. Here we report the successful extended immobilisation of coral larvae in a low-gelling temperature agarose. Embryos and larvae of five broadcast-spawning Scleractinian species were immobilised in agarose gel and tested in a series of exploratory survival and settlement assays. The optimal developmental stage for immobilisation was after ciliation at approximately 24 hours post-fertilisation, after which, survival of immobilised larvae of all species was nearly 100%. In long-term assays, 50% of Montipora digitata larvae survived immobilised for 89 days. Furthermore, immobilised larvae of multiple species, that were released from the agarose, generally remained capable of settlement. These results demonstrate that the immobilisation of the early life-history stages of corals is possible for a variety of applications in basic and applied science.

We determined the effect of acute and chronic toxicity of cadmium (Cd) on Pseudodiaptomus annandalei, according to their developmental and reproductive stages. Firstly, to estimate the 50% lethal concentration (LC50), acute exposure of nauplii and copepodids to 20, 40, 80, 150, and 300 μg/L of Cd was tested, and the effects of 5 μg/L and 40 μg/L of Cd on copepod developmental rate was done. Female lifespan and number of nauplii produced were compared. Secondly, one generation of copepod was exposed to dissolved (WCd) and dietary (DCd) Cd, and sex-specific Cd uptake and population density were estimated. 96 h LC50 was 40 μg/L Cd for nauplii and 120 μg/L Cd for copepodids. Duration of copepod development was 3.5 days and 5.5 days longer than the control when exposed to 5 μg/L and 40 μg/L of Cd, respectively. Female lifespan in both treatments were 9 and 8 days shorter than in the control, respectively. Total number of nauplii produced per female lifespan was 440 (control), 450 (5 μg/L Cd), and 365 (40 μg/L Cd). Cd uptake in copepods increased from nauplii to adults when exposed to dietary Cd and decreased when exposed to dissolved Cd. Dietary uptake of Cd was significantly higher in males than in females and Cd uptake from water was higher in males than in females, but not statistically significant. The total population of copepods were significantly affected by Cd. The toxic effects of Cd in copepods appear to depend on developmental stage, sex, duration and uptake route.

Aflatoxins are carcinogenic secondary metabolites produced mainly by two species of Aspergillus fungi, (A. flavus and A. parasiticus). The toxin contaminates maize grains during the developmental stage as well as in storage. Grains with contamination levels above 20 ng/g are usually destroyed in the USA while the European Union and Japan allow 2–4 ng/g and nil respectively. Ghana lacks regulatory infrastructure for monitoring fungal contamination and detection of aflatoxin in grains prior to market. Therefore, this study assessed the extent of A. flavus contamination together with other mould on maize and total aflatoxin accumulation among stored maize grains across 34 towns and villages under three agro-ecologies in Ghana during the minor crop season of 2015–2016. A. flavus, was identified as the most predominant contaminant and recorded average percentage contamination of 56.7%, 30.6%, 53.5% and 45.6% on grains sampled across four communities (Fumesua, Wenchi, Ejura and Akomadan), respectively. Total aflatoxins recorded in the samples per community were in the range of below limit of detection (LOD) to 692 ng/g, 23 ng/g, 945 ng/g and 112 ng/g for Fumesua, Wenchi, Ejura and Akomadan, respectively. These dangerous upper limits demand urgent attention in the area of aflatoxin resistance breeding in maize to help address the aflatoxin menace in Ghana.

Acanthopagruslatus and A.schlegelii (Sparidae) are closely related species distributed in northern tropical to temperate coastal waters of eastern Asia. Early life histories of the two species were investigated by seine net sampling in the Tien Yen estuary, northern Vietnam, which is probably the southernmost co-occurrence locality for these species. In northern Vietnam, early juveniles of both the Acanthopagrus species occurred in the Tien Yen estuary from December to March (A.schlegelii) or December to April (A.latus) when temperatures ranged from ca. 16 to 25 °C. The given temperature range is observed twice (spring and autumn) a year in Japan, and A.latus and A.schlegelii early juveniles are abundant respectively in each season. Thus, a thermoperiodicity of early juveniles of the two Acanthopagrus is common Asia-wide. The two species occurred as the same developmental stage, being markedly more abundant and having larger sizes in A.latus. The Acanthopagrus early juveniles inhabited bank waters of the estuary. Although waters continued to be mixed horizontally by temperatures but to be stratified by salinities during the study period, there was little variation in developmental stage and size among stations for both species. The two Acanthopagrus species from Vietnam spawn for the same season and their early juveniles simultaneously use the same nursery ground.

Anther culture is an important in vitro culture technique for production of double haploids. It is largely species and genotype-specific. Asian cultivated rice (O. sativa, ssp. indica) is recalcitrant to anther culture which limits its practical application in rice breeding. Several researchers tried to optimize the media recipes and culture techniques for callus induction and plantlet regeneration. Negligible response to callus induction and recovery of higher frequency of albino plants are the major hindrance for use of the technique in crop improvement. Shortening the culture period and sexual hybridization of rice sub-groups (japonica/indica) may be adopted to improve green plant regeneration from anther-derived callus in indica rice. However, genetic transformation technology using individual genes related to different aspects of anther culture could be a more direct approach. Besides, the role of genotypes, physiological status of donor plant, developmental stage of pollen, pre-treatment, culture media, phytohormones and culture conditions for successful anther culture have been discussed. Key words: Anther culture, factors for anther culture, double haploids, genetic improvement, rice.

The dichloromethane extraction was applied to extracted volatile compounds of the six developmental stages of caps and stipes of an Amanita rubescens mushroom and the relative contents were measured with the gas chromatography-mass spectrometry. The number of identified compounds ranged between 53 and 52, respectively, with a high ratio of alkane volatiles. The significant differences between the aroma compounds were determined in caps to identify their stages of development. The fully mature stage caps were characterized by 4,6-dimethyl-dodecane (7.69 ± 1.15%), 2-hexyl-1-decanol (11.8 ± 1.61%), 1,3-di-tert-butylbenzene (11.4 ± 1.25%), heptadecyl pentadecafluorooctanoate (2.16 ± 0.31%), and 2-hexyl-1-dodecanol (13.5 ± 1.33%). Niacinamide (3.90 ± 0.07%) and glycerol (3.62 ± 1.27%) was present in the caps in the early-stage of the rotting mushroom, which represented the 10th–12th day of fructification. The caps and stipes from the 12th–15th day of fructification were characterized by 2,3-butanediol (11.7 ± 0.13% and 8.00 ± 0.10%, respectively). Moreover, the caps from this developmental stage were characterized by 2-methyl- and 3-methyl butanoic acids (0.18 ± 0.03% and 0.33 ± 0.02%, respectively) which are typical for the rotting stage. In this study, we confirmed the effect of A. rubescens developmental stages on the aroma profile.

Thousands of microbial taxa in the soil form symbioses with host plants, and due to their contribution to plant performance, these microbes are often considered an extension of the host genome. Given microbial effects on host performance, it is important to understand factors that govern microbial community assembly. Host developmental stage could affect rhizosphere microbial diversity while, alternatively, microbial assemblages could change simply as a consequence of time and the opportunity for microbial succession. Previous studies suggest that rhizosphere microbial assemblages shift across plant developmental stages, but time since germination is confounded with developmental stage. We asked how elapsed time and potential microbial succession relative to host development affected microbial diversity in the rhizosphere using monogenic flowering-time mutants of Arabidopsis thaliana. Under our experimental design, different developmental stages were present among host genotypes after the same amount of time following germination, e.g. at 76 days following germination some host genotypes were flowering while others were fruiting or senescing. We found that elapsed time was a strong predictor of microbial diversity whereas there were few differences among developmental stages. Our results support the idea that time and, likely, microbial succession more strongly affect microbial community assembly than host developmental stage.

Densities of eggs and larvae of Engraulis anchoita and the nutritional condition of larvae were analyzed in the fixed sampling station (EPEA) in the period 2000-2017. These variables were analyzed seasonally and trends over time were determined. Ichthyoplankton samples were collected by oblique trawls using Bongo nets with 300 μm of pore mesh and fixed with formaldehyde 5% in seawater. Six morphometric variables were measured to estimate the nutritional condition of larvae. The developmental stage of each specimen was also determined. The highest mean value of egg density was detected in the October-December period, with a secondary mode in August, followed by one in March. Larval density presented a mode in October. Autumn and spring were the most favorable seasons for larval condition while winter proved to be the least favorable one. An increase in food availability during autumn and spring due to primary and secondary production peaks could explain the high nutritional condition and growth values observed in these seasons at the EPEA. No changes in trends of variables over time were detected. The integrated study of density and nutritional condition of E. anchoita larvae allows the determination of favorable breeding periods for the species, while the continuation of the long term study will allow evaluating possible effects of climate change in the early life stages of this species.

For intertidal marine species, salinity and temperature are important environmental factors with conditions ranging from oceanic salinity and temperature at high tide to almost freshwater conditions due to rain accompanied by aerial temperature exposure at low tide. The interactive effects of these stressors across development to the juvenile is not known. We investigated the impacts of salinity (29–41 ppt) and temperature (20°–24 ℃) on holobenthic intertidal development to the five-armed juvenile asterinid seastar Parvulastra exigua. There was an interaction between the stressors with highest mortality in the high salinity and temperature (38 ppt, 41 ppt, 24 ℃) treatments. Developmental stage progression was enhanced by increased temperature. On day 1, there was an interaction between salinity and temperature on developmental stage progression, where high salinity (41 ppt) retarded development at the control temperature (20 ℃). The juveniles reared at 24 ℃ were smaller than those at cooler temperatures, suggesting application of the temperature size rule. The low (29 ppt) and high (38 ppt, 41 ppt) salinity treatments produced the smallest and largest juveniles, respectively. In the 35 ppt and 38 ppt salinity conditions at 20 ℃, most juveniles had the normal five-armed profile. Body plan development was perturbed with deviations from pentamery at 29 ppt, 32 ppt and 41 ppt and at 22 ℃ and 24 ℃ in formation of no armed juveniles and ones with abnormal arm number (4, 6, 7 arms). Normal development at 35–38 ppt and 20–22 ℃ reflects breeding period conditions. Prolonged exposure to low and high salinity and increased temperature is deleterious to development in P. exigua and alters body morphology with implications for function and survival.

Onion thrips, Thrips tabaci Lindeman (Thysanoptera: Thripidae) is one of the most damaging insect pests of onions, Allium cepa L., which is an economically important agricultural crop cultivated worldwide. In this study, the combined application of entomopathogenic nematodes with entomopathogenic fungi against different soil dwelling stages of T. tabaci was evaluated. The nematodes included Heterorhabditis bacteriophora (VS strain) and Steinernema feltiae (SN strain), and fungi included Beauveria bassiana (WG-11) and Metarhizium anisopliae (WG-02), all four paired combinations (nematode + fungus) were included. In a small cup bioassay, only the combined application of H. bacteriophora and B. bassiana (WG-11) caused a synergistic interaction against pre-pupae, while all other combinations were compatible in an additive manner against pupae and late second instars. In a larger arena, a potted soil bioassay, again, combined applications of both pathogens produced greater mortality compared to single applications of each pathogen, all the combinations exhibited additive interactions, with the highest mortality observed in pre-pupae, followed by pupae and late second instar larvae using H. bacteriophora and B. bassiana (WG-11). Additionally, in the potted plant bioassay, lower adult emergence was observed from treated groups compared to control groups. Under field conditions, lower numbers of adults and larvae were found in treated groups relative to controls. Overall, the pre-pupal stage was more susceptible to the pathogen treatments, followed by pupae and late second instar larvae, and also combined applications of both pathogens suppressed the adult population. Combined application of entomopathogenic nematodes and fungi could be used for integrated pest management (IPM) of T. tabaci in onion production systems.

Chloris elata is an important weed for grain and sugarcane crops. In addition to its aggressiveness, it may show resistance to glyphosate herbicide. In this context, the aim of this study was to assess the effectiveness of glyphosate formulations, isolated or in association with ACCase inhibitors, in controlling C. elata (putatively resistant to glyphosate) at different developmental stages. Four experiments were conducted in a completely randomized design. Treatments consisted of glyphosate application under different formulations with ACCase inhibitors, isolated or in mixtures. Applications were carried out at the stages of four fully expanded leaves in Experiment I, four tillers in Experiment II, and at regrowth of the four-tiller plants in Experiments III and IV. Applications of glyphosate isopropylamine salt associated with sethoxydim or clethodim showed to be among the best treatments in Experiments I, II, and III, presenting control scores equal to or greater than 90%. However, not even these treatments could provide successful control in Experiment IV. Moreover, and regardless of the formulation, isolated glyphosate, showed 85% (Experiment III) and 50% (Experiment IV) maximum controls. Associations between glyphosate and ACCase-inhibiting herbicides showed to be effective in controlling C. elata, especially at early developmental stages. In general, isolated herbicides provided lower percentages of control, as well as higher values of dry matter. Sole herbicide applications were not effective in controlling C. elata (putatively resistant to glyphosate), regardless of the developmental stage. Highlights: Mixtures between glyphosate and ACCase-inhibiting have been shown to be effective in C. elata control, especially in the early development stages. Isolated herbicide applications were not effective in controlling C. elata, regardless of the development stage. Mixtures between glyphosate and ACCase-inhibiting is more important in the control than the glyphosate formulation or ACCase-inhibiting used.

Palm weevil ( Rhynchophorus phoenicis ) is an edible insect that is highly exploited as food. This study investigated variability in nutritional content during three developmental stages: early larva (ESL), late larva (LSL) and adult stage (ADS) and application of early larva stage in the formulation of complementary foods from the blend of broken rice and soybean flour at different proportions: A (100:0), B (94.5:5.5), C (89.5:10:5), (85.3:14.7), E (75.5:24.5) and F (70:30). Samples were assessed for proximate and mineral content while anti-nutritional factors and sensory analysis were conducted on the complementary food samples. Standard methods of analysis were used in all determinations. Results of proximate analysis indicated protein and lipid content were significantly different (p