Embryo Volume Research Articles

Influence of the shortened warming protocol on human blastocyst viability: an in vitro experimental study

Research QuestionDoes the shortened warming protocol impact the cell viability and outgrowth competence of human vitrified blastocysts warmed with or without fatty acids (FAs)? DesignIn this study, 326 discarded vitrified human blastocysts donated for research by consenting couples were used. The blastocysts were randomly allocated to five groups depending on the warming solutions, protocols, and recovery culture media: the control-conventional, control-shortened, FA-conventional, FA-shortened, and FA-shortened/recovery culture with FA (RF) groups. The blastocysts were warmed with or without FAs following the manufacturer's instruction (conventional method) or using the shortened method, in which blastocysts were immersed in a thawing solution for 1 min and then cultured in the recovery medium for 2 h. The embryo volume recovery, cell viability, intracytoplasmic lipid droplets, and outgrowth competence were evaluated. ResultsThe degree of blastocyst volume recovery was significantly higher after shortened warming than after conventional warming. Cell membrane collapse was observed during the shortened warming. The blastocyst survival rate and expansion status after the recovery culture were comparable among the five groups. However, the proportion of necrotic cells was increased in the control-shortened, FA-shortened, and FA-shortened/RF groups. The adhesion rates were comparable among all the groups; however, the outgrowth area was significantly higher in the FA-conventional group than in the other groups. ConclusionsThe efficacy of the shortened warming protocol depended on the warming solutions used. The shortened protocol effectively improves the daily workflow; however, each laboratory should examine its impact thoroughly before introducing it to prioritise clinical outcomes.

Reproductive traits of Gnathophylloides mineri Schmitt, 1933, a minute ectosymbiotic shrimp associated with the sea urchin Tripneustes ventricosus

Abstract Numerous studies on the reproduction of decapods have revealed that the female energy investment in embryo production is generally around 10% of the female weight. In decapods living together with other organisms, however, the reproductive output (RO) may be substantially higher. Here we tested the hypothesis that decapods living in association with other organisms allocate more energy in embryo production than free-living ones. We studied the shrimp Gnathophylloides mineri, which lives among the spines of the sea urchin Tripneustes ventricosus. Shrimps were collected between October 2011 and April 2013 along the southern Caribbean coast of Costa Rica. A total of 45 ovigerous females were analyzed; female size ranged from 1.44 to 2.14 mm carapace length. Females carried on average 37 ± 16 SD embryos. The embryo volume increased during the incubation period by 200% (from 0.04 ± 0.01 to 0.08 ± 0.01 mm3, mean ± SD). The RO was not related to female size, and comprised on average 27.5 ± 16.0 SD % of the dry weight of the females. In summary, G. mineri produces few and small embryos, but these embryos are relatively large when compared to similar-sized caridean species. Moreover, RO of this ectosymbiotic species was higher than in free-living species, but lower than in endosymbiotic decapods. These results support the hypothesis that energy investment in embryo production of decapods increases with the level of protection associated with the symbiosis.

Three-dimensional morphology of bovine blastocysts hatched against lipopolysaccharide exposure in vitro

Embryo transfer in cattle is globally becoming more ubiquitous, but the pregnancy rate is lower than that of artificial insemination. The uterus contains its own bacteria, and concentrations of lipopolysaccharides (LPS) from gram-negative bacteria are higher in uteri affected by endometritis than in healthy uteri and they suppress embryogenesis. The purpose of this study was to investigate the morphological characteristics of bovine embryos with a higher viability and implantability, by analyzing the morphology of bovine blastocysts that successfully hatched under challenge of LPS, using an optical coherence tomography (OCT) system. Developing embryos produced by in vitro fertilization that had reached the blastocyst stage on Day 7 were three-dimensionally scanned using an OCT system, then were continued to culture with or without LPS until Day 9, when the presence or absence of hatching was determined. The OCT-captured three-dimensional images were used to quantify 20 different metrics, including inner cell mass (ICM), trophectoderm, blastocoel, and total embryo volume; each of the parameters was compared between the hatched and unhatched embryos. Under the LPS challenge, hatched embryos had higher ICM thickness and volume, and lower trophectoderm thickness than unhatched embryos. Furthermore, hatched embryos under LPS challenge had higher ICM thickness and ICM volume than hatched embryos without LPS challenge. The present results suggest the possibility that ICM thickness and ICM volume calculated by OCT system could be indices for good quality bovine embryos.

Reproductive biology aspects with a specific focus on histological analysis of oogenesis in Heterocarpus woodmasoni Alcock, 1901 from Southeastern Arabian Sea, Indian Ocean

The expansion of deep-sea fisheries in India has underscored the necessity for comprehensive data on the reproductive characteristics of targeted species to facilitate effective resource management. Heterocarpus woodmasoni Alcock, 1901 plays a pivotal role in supporting India’s deep-sea shrimp fishery sector. In this pioneering study, we provide novel insights into the reproductive dynamics of H. woodmasoni, utilizing specimens collected from 2019 to 2022 at the primary deep-sea shrimp landings site of Sakthikulangara fisheries harbour, Kerala, India. The research revealed that H. woodmasoni exhibits a continuous breeding pattern characterized by three prominent spawning peaks: two occurring in January and April with an additional extended peak observed in September. Moreover, the study observed a consistently high proportion of ovigerous females throughout all seasons, suggesting year-round reproductive activity in H. woodmasoni. Size (CL) at the onset of sexual maturity (SOM) for males and females measured 16 mm and 17 mm, respectively. In terms of brood size, the mean absolute brood size estimated in H. woodmasoni was 5438 ± 2687. Interestingly, while brood size increased as a power function of carapace length, the correlation between reproductive output and carapace length was found to be statistically not significant. Embryo volume varied significantly between the three developmental stages (Stage I: 0.03 ± 0.02 mm3, stage II: 0.05 ± 0.04 mm3 and stage III: 0.11 ± 0.03 mm3). The gonadosomatic index showed increasing trend corresponding ovary maturation. However, the index declined in the final maturity stage characterized by a black coloured ovary. The vivid colour changes observed in mature ovaries, spanning early vitellogenesis, late vitellogenesis and maturation phases offer valuable insights into the dynamic cellular transformations occurring during ovarian development. The present study regarding the various reproductive traits of H. woodmasoni with histological examination of oogenesis will help to facilitate sustainable exploitation and adequate conservation management of this species.

The relationship between grade and metabolism for in vivo and ICSI derived embryos

The objective of this study was to describe the metabolism of glucose, pyruvate and lactate by in vitro- (ICSI derived) and in vivo-derived equine embryos and the relationship with embryo grade and sex. During the course of this study, a simplified embryo grading system was developed for both in vivo- and in vitro-derived embryos that proved to be significantly associated with both embryo metabolism and survival rates following transfer into a recipient mare. A total of 15 in vivo derived embryos and 32 ICSI-derived embryos were cultured in protein free synthetic oviduct fluid (SOF) for 24h between day 8 and day 9 of development. Aliquots of 10µL of media before culture (t=0) and spent media after culture (t=24h) were snap frozen for subsequent analysis by GC-Mass Spectrometry for glucose, lactate and pyruvate. The embryos were photographed and classified as grade 1, 2 or 3 based on the percentage of extruded debris between the trophoblast and the zona pellucida. In this study the sex of the embryos (in vivo n = 5, ICSI n = 25) was determined by PCR. At day 8 of development, the embryo metabolism of glucose, pyruvate and lactate in relation to embryo origin (in vivo vs in vitro), grade and sex wasevaluated following 24h of culture in protein free media. Overall, there was a positive correlation between embryo volume and lactate peak area (r2 = 88.2%, p ≤ 0.001) and a negative correlation with both glucose peak area (r2 = 29.8%, p =0.023) and pyruvate peak area (r2 = 46.2%, p =0.003) in the spent media. These results illustrated that as embryo volume increased, the amount of glucose and pyruvate in the media decreased and the production of lactate increased. After accounting for embryo volume, general linear model analysis revealed an effect of embryo grade (p =0.031), but no significant effect of embryo sex or origin on lactate consumption over 24h. Grade 2 embryos, however, consumed more lactate than Grade 1 embryos (p = 0.029). In conclusion, the metabolic relationships between glucose, lactate and pyruvate were described for both in vivo- and in vitro-derived equine embryos in this study. Understanding the optimal metabolic conditions for in vitro-derived embryos in comparison with in vivo-derived embryos will enable the development of models to investigate early embryonic death and to optimize pregnancy rates in the mare.

Expert consensus on the morphological evaluation of human cleavage-stage embryos and blastocysts.

Expert consensus on the morphological evaluation of human cleavage-stage embryos and blastocysts.

Variation in body size and reproductive characteristics among chromosomal races of the Sceloporus grammicus complex in México

Abstract Populations and chromosomal races of a species often show variation in life-history characteristics due to phylogeny and environmental factors. We analysed variation in snout–vent length (SVL) and reproductive variables, such as minimum SVL at sexual maturity, litter size and embryo volume in females of the Sceloporus grammicus lizard complex, which includes seven chromosomal races as well as hybrid zones among different combinations of races (F5, F5 + 6, potential but unverified F5 + 6, F6, FM1, FM2, FM2 × F5 hybrid zone, FM3, LS and unverified SGG). The SGG, F5 and LS females were larger than F6 and FM3 females. Females with non-vitellogenic follicles and vitellogenic follicles were smaller than females with embryos. The SVL of females containing embryos varied among chromosomal races, the largest being the same as indicated above, but the smallest was the FM2 × F5 hybrid zone. Litter size was similar among chromosomal races (5.8 embryos), while embryo volumes varied among them. Two patterns of reproductive activity were found: seasonal and continuous reproduction. Our results showed the influence of environmental factors of each locality, which supports the ecological hypothesis. The use of chromosomal races did not reveal a significant phylogenetic effect, because allozyme and molecular data show only small pairwise genetic distances among the S. grammicus populations on the Mexican Plateau.

DNA barcoding, dwelling morphology, and fecundity of the gall-forming shrimp Paratypton siebenrocki Balss, 1914 (Caridea: Palaemonidae)

ABSTRACT Tropical coral reefs offer a wide variety of habitats to countless invertebrate species. Sessile host organisms especially are inhabited by small taxa, of which decapod crustaceans form one of the most diverse communities. Symbiotic palaemonid shrimp species associate with marine invertebrate hosts from multiple phyla, including cnidarians such as stony corals (Scleractinia). The intriguing gall-forming shrimp Paratypton siebenrocki, a symbiont of Acropora corals in the Indo-Pacific, was collected in the Saudi Arabian Red Sea, Kenya, and the Maldives. Based on morphology P. siebenrocki has been considered to be most closely related to the genera Anapontonia and Metapontonia; however, no clear clustering with either palaemonid genus was observed in a phylogenetic reconstruction based on 16S and COI mtDNA. Here we photo-document the dwellings of P. siebenrocki in Acropora spp. for the first time, and furthermore we report on the reproductive output of this species. The number of eggs ranged from 345 to 909 (n = 6), and embryo volume differed strongly between early- and late-stage embryos. The carapace length ranged from 2.58 to 4.55 mm for the females and 1.51 to 2.5 mm for the males (n = 5). The number and size of the embryos, combined with their specialised, secluded lifestyle, suggest that P. siebenrocki allocates higher energy towards embryo production than free-living confamilials do.

Retracted: The Clinical Value of 3D Ultrasonic Measurement of the Ratio of Gestational Sac Volume to Embryo Volume in IoT-Based Prediction of Pregnancy Outcome.

[This retracts the article DOI: 10.1155/2021/6421025.].

Microcystin-LR inhibits early pregnancy by impairing the vascular network of luteum: Involvement of the MEK/ERK/SP1/VEGFR2 axis.

Microcystin-leucine-arginine (MC-LR) reduces the fertility of female mice, but the mechanism is unknown. We studied the effect of MC-LR on early pregnancy and elucidated its possible mechanism. The number of embryo beds and embryo volume decreased in pregnant mice at 6 or 8 days after fertilization after acute exposure to MC-LR. The corpus luteum secretes estrogen and progesterone, which are involved in embryo implantation and maintenance of early pregnancy. MC-LR exposure reduced luteal blood vessel branches and inhibited hormone synthesis. Functional blood vessels are essential to the maintenance of luteal structure and function. Reduced migration and tube-forming were also detected in human umbilical vascular endothelial cells (HUVECs) treated with MC-LR. MC-LR significantly decreased the expression of vascular endothelial growth factor receptor 2 (VEGFR2) in vivo and in vitro, which was responsible for the inhibited construction of the vascular network. The MEK/ERK/SP1 signal pathway mediated the decrease in VEGFR2 expression, and the agonists of phosphorylated extracellular regulated protein kinases (p-ERK) alleviated the anti-angiogenic effect of MC-LR. In conclusion, we demonstrated the toxicity of MC-LR on construction of vascular network in corpus luteum, which could provide a new perspective on female infertility or miscarriage caused by environmental factors.

Vanadium Modulates Proteolytic Activities and MMP-14-Like Levels during Paracentrotus lividus Embryogenesis.

The increasing industrial use of vanadium (V), as well as its recent medical use in various pathologies has intensified its environmental release, making it an emerging pollutant. The sea urchin embryo has long been used to study the effects induced by metals, including V. In this study we used an integrated approach that correlates the biological effects on embryo development with proteolytic activities of gelatinases that could better reflect any metal-induced imbalances. V-exposure caused morphological/morphometric aberrations, mainly concerning the correct distribution of embryonic cells, the development of the skeleton, and the embryo volume. Moreover, V induced a concentration change in all the gelatinases expressed during embryo development and a reduction in their total proteolytic activity. The presence of three MMP-like gelatinases (MMP-2, -9, and -14) was also demonstrated and their levels depended on V-concentration. In particular, the MMP-14-like protein modified its expression level during embryo development in a time- and dose-dependent manner. This enzyme also showed a specific localization on filopodia, suggesting that primary mesenchyme cells (PMCs) could be responsible for its synthesis. In conclusion, these results indicate that an integrated study among morphology/morphometry, proteolytic activity, and MMP-14 expression constitutes an important response profile to V-action.

Real-time computer assisted measurement of oocyte and embryo volume for assessment of transport parameters

Cryopreservation of gametes has revolutionized both animal agriculture and human reproductive medicine. Although many new technologies have tremendously improved the cryopreservation of oocytes and embryos, osmotic stress encountered during the equilibration process can cause their loss of function. Rational cryoprotective agent (CPA) equilibration strategies can be used to minimize this stress but require trained personnel to monitor the process in individual oocytes or embryos or require the use of suboptimal average transport parameter values in mathematically guided protocols. To enable individually optimized equilibration of CPAs in individual cells, here we establish experimental and computational techniques to track the osmotic behavior of individual bovine oocytes and embryos during CPA equilibration in real time. We designed a microfluidic device to provide a controlled flow of CPA and modified standard image analysis techniques to estimate real-time cell volume changes. In particular, we used a level-set method to define a boundary within a contour plot which could automate the image analysis process. A colour based level set algorithm coupled with contour smoothing not only provided the best fit but also reduced the segmentation time to well under a second per image. The accuracy of the automated method was comparable to human segmented images for both oocytes and embryos. This technology should enable both rapid evaluation of key biophysical parameters in oocytes and embryos undergoing CPA equilibration and the development of real-time feedback-control of CPA equilibration, enabling individual oocyte- and embryo-specific optimal protocols.

A molecular dynamics study on the mechanism of heterogeneous bubble nucleation of mixed liquid

The nucleate boiling of mixed boiling is widely utilized in natural gas, hydrogen liquefaction process, and other fields. In this paper, the molecular dynamics study of heterogeneous bubble nucleation of pure argon liquid and the argon‑krypton mixed liquid on the platinum surface was conducted. The nucleate boiling characteristics were presented and the variations of the density, temperature, nucleation embryo volume, and heat flux were analyzed. The difference in the nucleation mechanism of pure liquid and mixed liquid was explained based on the competition of atomic potential energy and atomic kinetic energy. The effect of the component mole fraction on the bubble nucleation of mixed fluid was also discussed. The results showed that the onset superheat of boiling of argon‑krypton mixed liquid is higher than that of pure argon. The higher the mole fraction of the component with a lower saturation temperature, the lower the onset superheat of the liquid nucleate boiling. When x < 0.5, the growth rate of the nucleation embryo is dominated by the x value, when x > 0.5, the growth rate is dominated by wall temperature. Moreover, the difference in diffusion coefficients of each component is one of the direct causes of determining the growth rate of nucleation embryos. The findings provide insights into the mechanisms of differences in boiling nucleation between mixed liquid and pure liquid, as well as guide the reasonable utilization of mixed liquid.

Reproductive Variability in Hippolytid Shrimp Shape Morphotypes

Shape morph-specific studies in hippolytid shrimps revealed significant results on their ecomorphology and evolutionary adaptations. Among the species of the genus Hippolyte, only one exhibits an unusual, sharp rostral dimorphism and has been used as an animal model for the investigation of mechanisms of the morph-specific adaptation: the intertidal Hippolyte sapphica. The species is endemic of the Central/Eastern Mediterranean basin and exhibits morph-A with a long dentate rostrum and morph-B with a short, juvenile-like one. The two morphotypes were recently confirmed to be conspecific, while offspring and morphological studies showed significant microevolutionary adaptations, which balance the disadvantage of the “rostral loss.” The present study aims to investigate the effect of such phenotypic variation on the reproductive traits of the species. We collected ovigerous females of H. sapphica in mixed (morph-A and morph-B) and unmixed populations (morph-A) along the species geographical range. We measured seven morphometric and maternal investment traits: carapace length, fecundity, embryo volume, egg density, female dry weight, brood dry weight, and reproductive output. Our results showed that ovigerous females were bigger in morph-A than in morph-B, whereas fecundity did not show any significant differences between the two morphotypes. High egg volume might be attributed to the latitudinal differences of our sampling sites compared to congenerics. Interestingly, the reproductive output was found to be bigger in morph-A specimens, suggesting that the maternal energy investment is selectively determined from the rostral presence/absence and the morphotype’s higher viability in the species populations.

P-190 The first cleavage plane in human embryos is dictated by the topology of pronuclei, not by the polar body position

Abstract Study question Is the first cleavage plane pre-determined in human embryos? Summary answer Unlike in mice, the human zygotes tend to divide at the right angle to the longitudinal pronuclear axis, irrespective of polar body position. What is known already In lower species, the orientation of the first cleavage plane defines the development of individual blastomeres and the three-dimensional (3D) body plan. The existence of embryonic pre-patterning in mammals remains controversial. Mouse studies suggested that the juxtaposed pronuclei (PN) are aligned with the polar body (PB) and cleavage furrow bisects the zygote along the polar axis thus ensuring symmetrical distribution of maternal resources to arising blastomeres. However, whether the same spatial control applies to humans is unclear. Study design, size, duration Retrospective observational analysis of high-resolution footage from 300 human embryos that gave rise to live births after a single embryo transfer. Participants/materials, setting, methods Embryo monitoring has been performed by the GERI time-lapse incubator. Image acquisition of individual embryo development lasted 6 days, starting within 10 minutes after intracytoplasmic sperm injection (ICSI). Transmitted light images have been automatically captured in 5-minute intervals and 11 focal planes covering the whole embryo volume. Analysis of 3D image data was performed at the time frame corresponding to the onset of the first mitosis. Main results and the role of chance The systematic analysis of the zygotic division pattern showed that the PN axis (defined by apposed pronuclei) was not stereotypically aligned with the animal-vegetal (AV) axis (with animal pole defined by the PB). Instead, PN/AV arrangement categories were equally represented in our cohort (PN/AV angle 0-30° - 34.7% vs. 30-60° - 36.0% and 60-90° - 29.3%; chi-square test p=.91351). Remarkably, we observed that the direction of the first cleavage was preferentially perpendicular to the PN axis (65.3%), while only 2.7% of embryos divided along the longitudinal PN axis. The remaining 32% embryos showed intermediated phenotype with cleavage plane/PN axis angle 30-60o. The results of this human embryo analysis contrast with the previous reports from mice. We hypothesize that this discrepancy might be attributed to the difference in centrosome inheritance and configuration of the zygotic spindle apparatus. Limitations, reasons for caution All analyzed embryos were produced by ICSI. Deposition of spermatozoa might affect the pronuclei positioning. Nevertheless, preliminary data from conventional IVF cycles are in agreement with the ICSI group results. Wider implications of the findings Our observations challenge the textbook view of the first division’s geometry, which has been derived from experiments in mice. The analysis of clinical time-lapse records improves our understanding of symmetry breaking during normal human development and creates scientific ground for evidence-based (de)selection of embryos for infertility treatment. Trial registration number not applicable

Chronic dietary exposure to a glyphosate-based herbicide results in reversible increase early embryo mortality in chicken

Glyphosate (Gly) is the active molecule of non-selective herbicides used in conventional agriculture. Some evidence shows that exposure to Glyphosate-Based Herbicides (GBH) can affect both male and female fertility in animal models. However, few data exist on birds that can be easily exposed through their cereal-based diet. To our knowledge, there are no current studies on the effects of chronic dietary exposure to GBH and the potential reversibility on the fertility and embryo development in chickens. In our protocol, hens (32 weeks-old) were exposed to GBH (47 mg kg-1/day-1 glyphosate equivalent corresponding to half of the No-Observed-Adverse-Effect-Level (NOAEL) as defined by European Food Safety Authority in birds, GBH group (GBH), n = 75) or not (Control group (CT), n = 75) for 6 weeks. Then, both CT and GBH groups were fed for 5 more weeks without GBH exposure. During these two periods, we investigated the consequences on the egg performance and quality, fertilization rate, embryo development, and viability of offspring. Despite the accumulation of Gly and its metabolite aminomethylphosphonic acid (AMPA) in the hen blood plasma, the body weight and laying rate were similar in GBH and CT animals. We observed from the 4th day of exposure an accumulation of Gly (but not AMPA) only in the yolk of the eggs produced by the exposed hens. After artificial insemination of the hens followed by eggs incubation, we showed a strong significant early embryonic mortality level in GBH compared to CT animals (78 ± 2 % vs 2.5 ± 0.3 %, p < 0.0001) with embryo death mainly occurring on the third day of incubation. By using computed tomography (CT) and magnetic resonance imaging (MRI) tools, we noted a significant delay in the embryo development of GBH survivors at 15 days with a reduction by half of the embryo volume and some disturbances in the calculated volumes of the embryonic annexes. At 20 days of incubation, we showed a reduction in the length of the tibia and in the volume of the soft tissues whereas the skeleton volume was increased in GBH chicks. The vast majority of these phenotypes disappeared two weeks after an arrest of the GBH maternal dietary exposure. Taken together, the dietary chronic exposure of broiler hens to GBH at a Gly equivalent concentration lower than NOAEL induces an accumulation of Gly in the egg yolk resulting in severe early embryonic mortality and a delayed embryonic development in survivors that were abolished two weeks after the end of GBH exposure.

How Egg Storage Duration Prior to Incubation Impairs Egg Quality and Chicken Embryonic Development: Contribution of Imaging Technologies.

Storing fertilised eggs prior to incubation is a frequent practice in commercial hatcheries to coordinate activities and synchronise hatchings. However, the conditions used to store eggs can have major impacts on egg quality and the subsequent viability of chicken embryos. While storage temperatures of 16–18°C are classically used in hatcheries, the duration of storage varies from three to more than 10 days. We explored the effect of storage duration (zero, three or 10 days; D0, D3 and D10, respectively) at 16°C, 80% relative humidity (RH) on egg quality (Broiler, Ross 308), using computed tomography (CT) and classical measurements (egg weight, eggshell strength, egg white pH, Haugh units, yolk index and colour). The results revealed that a storage duration of up to 10 days negatively affected some egg quality traits (yolk index and volume, air chamber volume and egg white pH). Eggs stored for three or 10 days were further incubated for 11, 13 or 15 days (37.8°C, 55% RH). Eggs were analysed by magnetic resonance imaging (MRI) and CT to assess the development of the embryo and internal egg changes occurring during incubation. First, data showed that the fertility and sex ratio of eggs were not affected by storage duration. However, the mortality of viable eggs was increased in the D10 group compared to the D3 group. Results of non-invasive imaging technologies revealed that the storage of eggs for 10 days impaired embryo growth as early as 11 days of incubation (decrease in brain and embryo volumes). Collectively, these data provide new evidence that the duration of egg storage negatively affects embryonic growth. They further corroborate that this parameter is likely to be crucial to synchronising embryonic stages and maybe reducing the hatching window, hence limiting the time spent by newborn chicks in hatchers. In addition, our results highlight that CT and MRI imaging technologies are useful non-invasive tools to evaluate egg quality prior to incubation and the impact of storage (or incubation) practices on developmental growth of the embryo.

Preliminary studies on variation in the micromorphology of the seed coat inHabenaria s.l.(Orchidaceae) and relatives

AbstractThere are many studies on the biology, ecology and infrageneric classification of the cosmopolitan orchid genus Habenaria. However, there are few data on the seed morphology of species in this genus. The aim of this study was to analyse the seed morphology of Habenaria and related taxa (111 species in total). The species of Habenaria s.l. studied are mostly characterized by a scobiform seed, but the form of individual seed coat cells varies between and among some species. In most of the species analysed, the cell walls are arranged mostly along the main axis of a seed. The size of the embryos and the volume of seeds and embryos also varied in Habenaria. The periclinal wall of the cells in the seed coat of most of the taxa examined is smooth, but there are several species with distinct ornamentation on this wall (slanting ridges, transverse ridges, reticulate ridges). The periclinal wall could be used in taxonomic delimitations, especially when combined with additional features such as seed size, shape and arrangement of the cells in the seed coat. Although many of the morphological groups in Habenaria s.l. were included in this study, no qualitative seed characters that could be helpful in establishing an alternative infrageneric division of this taxon were previously recognized.

Application of X-ray computed tomography to analyze the structure of sorghum grain

BackgroundThe structural characteristics of whole sorghum kernels are known to affect end-use quality, but traditional evaluation of this structure is two-dimensional (i.e., cross section of a kernel). Current technology offers the potential to consider three-dimensional structural characteristics of grain. X-ray computed tomography (CT) presents one such opportunity to nondestructively extract quantitative data from grain caryopses which can then be related to end-use quality.ResultsPhenotypic measurements were extracted from CT scans of grain sorghum caryopses. Extensive phenotypic variation was found for embryo volume, endosperm hardness, endosperm texture, endosperm volume, pericarp volume, and kernel volume. CT derived estimates were strongly correlated with ground truth measurements enabling the identification of genotypes with superior structural characteristics.ConclusionsPresented herein is a phenotyping pipeline developed to quantify three-dimensional structural characteristics from grain sorghum caryopses which increases the throughput efficiency of previously difficult to measure traits. Adaptation of this workflow to other small-seeded crops is possible providing new and unique opportunities for scientists to study grain in a nondestructive manner which will ultimately lead to improvements end-use quality.

New Insights into the Life-history Traits of the Night Shrimp Processa hemphilli (Caridea: Processidae) in the Western Atlantic.

We analyzed sexual dimorphism, sex ratio, relative growth, morphological sexual maturity, fecundity, embryo volume, and reproductive output of a population of the night shrimp Processa hemphilli from the Southeast coast of Brazil. Sampling was seasonally conducted, from June 2016 to April 2019 in Ubatuba, São Paulo. Our results showed the presence of sexual dimorphism in the body size, with females being larger than males. In addition, there was no difference in the overall sex ratio. However, it was possible to observe a skewed sex ratio to males in the smaller size classes, whereas females were more abundant in the larger size classes. The estimated carapace length at the onset of morphological sexual maturity of females and males was 3.6 mm and 3.2 mm, respectively. The mean fecundity of females was 285 ± 142 embryos (mean ± SD) embryos and was positively correlated with the carapace length. The embryo volume differed between the developmental stages with a constant increase in volume throughout the development between stages I and III. The mean reproductive output of females was 52.4%. Our results suggest that P. hemphilli have a pure search mating system, show sex allocation in the growth of body structures, and that females have high reproductive investment. All of these results represent the first insights into the life-history traits of the species in the western Atlantic and contribute to the knowledge about processid species.