The intra-ovarian presence of ootids, i.e. female gametes that have completed meiosis, is considered exceptional in the animal kingdom. The present study explores the first such case to be reported in a sea cucumber (Echinodermata: Holothuroidea). In the overwhelming majority of animals, including holothuroids, oocytes (i.e. immature female gametes) that are developing in the ovary undergo a primary arrest at the prophase stage of meiosis, which may last from days to decades. In free-spawning taxa, this arrest is normally lifted only during or shortly before transit in the gonoduct, when gamete release (spawning) is imminent. However, oocytes of the holothuroid Chiridota laevis were discovered to have resumed the second meiotic division including the completion of germinal vesicle breakdown and polar-body expulsion inside the ovary, effectively reaching the ootid stage concomitantly with ovulation (i.e. escape from follicle cells) prior to spawning. The potential drivers and significance of this exceptionally rare case of full intra-ovarian oogenic maturation are discussed.

The reproductive cycle of Marphysa sanguinea is described for a population at Mount Edgecombe, Plymouth, near the type location in Southwest England, using a data set obtained previously (October 1999 to September 2000). The species is iteroparous without schizogamy, spawning prior to October 1999 and during a short breeding season in 2000 from end August through September. The sexes are separate with a sex ratio of 1:1. Mature oocytes and spawned eggs are 215 μm in diameter and spermatozoa of the ectaqua sperm type. Mature gametes of both sexes are discharged through paired coelomoducts, and the diploid chromosome number is 28. Proliferation of new coelomic gametes from paired gonads began within a month of spawning and continued for 8–9 months but ovulation was suppressed in June and July. Attempts to undertake fertilisation using spawned oocytes and active spermatozoa were unsuccessful. The size of discharged oocytes suggests a short pelagic larval duration of a few days. This is the first publication about the reproduction of this species, and our results suggest that M. sanguinea is restricted to intertidal areas in SW England, NW France and southern North Sea. The highly synchronised pattern of reproduction observed is not compatible with a quasi-cosmopolitan species range indicating that this species has been mistakenly reported from around the world. Future studies of the genus should combine rigorous taxonomy with observations of reproduction to facilitate comparison among Marphysa spp.

The present study described the gametogenesis and assessed the pattern of energy storage throughout the reproductive cycle of the golden carpet shell (Polititapes aureus) from the Ria Formosa lagoon (southern Portugal). Monthly sampling was performed for two years (March 2016–February 2018) and the study was based on gonad histology, complemented by the estimation of the mean gonadal index, body condition index and biochemical composition. The species’ reproductive cycle presented a seasonal pattern, with a resting period mainly between October and January and gametogenesis beginning around February–March. The spawning season of P. aureus was shorter in 2016 (June–September) than in 2017 (May–October). Furthermore, ripe individuals were very scarce (1.3%) in 2017 compared with the previous year (11.4%). Mean gonadal index (GI) reflected the species’ reproductive cycle and the body condition index (CI) and biochemical composition of the individuals exhibited high variation between years. Regarding the biochemical composition, proteins ranged between 190.6 and 595.2 μg mg−1, glycogen from 5.7 to 102.6 μg mg−1 and total lipids between 31.6 and 80.7 μg mg−1. The reproduction of P. aureus was strongly influenced by fluctuations in both seawater temperature and chlorophyll a, as reflected through the temporal variation in the gonadal cycle, condition index and biochemical composition. Overall, the information gathered in this study is valuable to propose scientifically based harvesting management measures for the long-term sustainable exploitation of this shellfish resource, further reinforcing the importance of implementing adaptive fishery management strategies to cope with global climate change.

Coccidia display a characteristic life cycle, where the parasites switch between asexual and sexual development, resulting in an environmental stage, the oocyst. The entero-pathogenic Cystoisospora suis, a coccidian parasite of swine and close relative to Toxoplasma gondii, undergoes development in one host-cycle. Despite the well-described intracellular development of Coccidia, the C. suis life cycle can progress in an in vitro, host cell-free system after initial intracellular development of merozoites. A novel host cell-free cultivation method was developed by transferring purified merozoites from cell culture supernatant (dpi 6) to culture medium and incubating them for 5 days to induce their progression to sexually differentiated stages. The development of sexual stages in the absence of host cells was verified by morphological studies, flow cytometry and the transcription analysis of three genes linked to sexual stages (HAP2, OWP and TyRP). The host cell-free culture permits the sexual development (and with this, the complete life cycle progression from sporozoites to oocysts) of C. suis in vitro and provides a new tool for detailed research on the development of C. suis and possibly other Coccidia. This will also be useful for the evaluation of novel drug or vaccine targets in these parasites.

Germ plasm-related structures (GPRS) are known to accompany meiotic cell differentiation but their dynamics are still poorly understood. In this study, we analyzed the ultrastructural mechanisms of GPRS transformation during oogenesis and spermatogenesis of the bivalve mollusc Ruditapes philippinarum (Manila clam), exploring patterns of GPRS activity occurring at meiosis onset, sex-specific difference/similarity of such patterns, and the involvement of mitochondria during GPRS-assigned events. In the two sexes, the zygotene–pachytene stage of meiosis is anticipated by three shared steps. First, the dispersion of germ plasm granules containing the germ line determinant VASA occurs. Second, the VASA protein deriving from germ plasm granules enters neighbouring mitochondria and appears to induce mitochondrial matter release, as supported by cytochrome B localization outside the mitochondria. Third, intranuclear VASA entrance occurs and the protein appears involved in chromatin reorganization, as supported by VASA localization in synaptonemal complexes. In spermatogenesis, these three steps are sufficient for the normal course of meiosis. In oogenesis, these are followed by the action of ‘germ plasm granule formation complex’, a novel type of structure that appears alternative to the Balbiani body. The possibility of germ plasm involvement in reproductive technologies is also suggested.

The life history traits of bathydraconids, deep-living fishes distributed all around the Antarctic continent, are poorly known. In particular, very few data are available on the relatively rare genera Akarotaxis and Bathydraco. With the aim to fill this gap, sagittal otoliths and gonads were analysed to assess individual age and reproductive features of Akarotaxis nudiceps (Waite, 1916), Bathydraco macrolepis Boulenger 1907 and Bathydraco marri Norman, 1938 collected in the Weddell Sea. Based on the annual growth increment patterns, age estimates ranged between 6–11, 5–11 and 8–11 years for A. nudiceps, B. macrolepis and B. marri, respectively. Most of the gametogenetic processes could be described based on gonad histology for both sexes. Females shared the reproductive features commonly reported in notothenioids, such as group-synchronous ovary development and prolonged gametogenesis. Total fecundity estimates were comparable between the two species of Bathydraco (1500–2500 eggs/female), whereas that of Akarotaxis was one order of magnitude smaller (200–250 eggs/female). Consistently, the mean size of late vitellogenic oocytes showed an opposite trend, being 1.6–1.8 mm in Bathydraco and 2.2 mm in Akarotaxis.

Glyphosate treatments to glyphosate-resistant (GR) cotton have been associated with poor pollination and increased boll abortion. Anatomical studies were conducted to characterize the effect of glyphosate treatments on the development of male and female reproductive organs of cotton flowers at anthesis. In comparison with nontreated plants, glyphosate applied at both the four-leaf stage postemergence (POST) and at the eight-leaf stage POST directed inhibited the elongation of the staminal column and filament, which increased the distance from the anthers to the receptive stigma tip by 4.9 to 5.7 mm during the first week of flowering. The increased distance from the anthers to the stigma resulted in 42% less pollen deposited on stigmas of glyphosate-treated plants than in nontreated plants. Moreover, pollen from glyphosate-treated plants showed numerous morphological abnormalities. Transmission electron microscopy showed the presence of large vacuoles, numerous starch grains, and less organized pockets of the endoplasmic reticulum containing fewer ribosomes in pollen from glyphosate-treated plants than from nontreated plants. Pollen development in glyphosate-treated plants is likely inhibited or aborted at the vacuolate microspore and vacuolate microgamete stages of microgametogenesis, resulting in immature pollen at anthesis. Although stigmas from glyphosate-treated plants were 1.2 to 1.4 mm longer than those from nontreated plants, no other anatomical differences in stigmas were visibly evident. The presence of the GR 5-enolpyruvylshikimate-3-phosphate synthase (CP4-EPSPS) enzyme from Agrobacterium sp. strain CP4 was quantified in reproductive and vegetative tissues using enzyme-linked immunosorbent assay. The content of CP4-EPSPS in the stigma, anther, preanthesis floral bud (square), and flower petals was significantly less than that in the vegetative leaf tissue. Glyphosate effects on the male reproductive development resulting in poor pollen deposition on the stigma, as well as production of aborted pollen with reduced viability, provide a likely explanation for reports of increased boll abortion and pollination problems in glyphosate-treated GR cotton.

Glyphosate treatments to glyphosate-resistant (GR) cotton can cause increased fruit loss compared with untreated plants, likely due to reductions in pollen viability and alterations in floral morphology that may reduce pollination efficiency. This study was conducted to determine whether both stamen and pistil are affected by glyphosate treatments by measuring seed set from reciprocal reproductive crosses made between glyphosate-treated GR, untreated GR, and conventional nontransgenic cotton. Pollen viability was 51 and 38% lower for the first and second week of flowering, respectively, in GR plants treated with a four-leaf postemergence (POST) treatment and an eight-leaf POST-directed treatment of glyphosate than in GR plants that were not treated. Seed set per boll was significantly reduced when the pollen donor parent was glyphosate treated vs. untreated for the first 2 wk of flowering. There were no significant differences between treatments applied to male parents as measured by seed set at Weeks 3 and 4 of flowering. Seed set was not influenced by glyphosate treatments applied to female parents at any time. Retention of bolls resulting from crosses was reduced by glyphosate treatment of male parents during the first and third week of flowering but was not affected by glyphosate treatment of female parents. The application of gibberellic acid (GA), which has been shown to reverse male sterility in tomato (Lycopersicon esculentum L.) and to enhance boll retention in cotton, was investigated for similar effects in glyphosate-treated GR cotton. The GA treatments to glyphosate-treated plants increased the anther–stigma distance 12-fold, stigma height, and pollen viability in the second week of flowering but decreased the number of seeds in second-position bolls on Fruiting branches 1 through 3, decreased the number of first-position bolls per plant, and increased the number of squares in comparison with glyphosate-treated GR plants not receiving GA. Although GA applications to glyphosate-treated GR cotton have some remedial effect on pollen viability, the GA-induced increase in the anther–stigma difference exacerbates the increase in anther–stigma distance caused by glyphosate, resulting in low pollination.

Endocrine disruption has rarely been reported in field populations of the edible cockle and the context with the general health of the shellfish is unclear. This study examined the reproductive state of two Cerastoderma edule populations over a 6-month period to assess their reproductive condition, the incidence of intersex and presence of parasitic infection. A further seven native sites from south-west England were examined during the peak reproductive season to identify the presence of intersex within the region. Laboratory exposures of organisms collected from field populations showed a significantly female-biased sex ratio compared with controls when exposed to the endocrine disrupting chemicals, bisphenol-A (nominal concentration: 0.1 µg L−1) and 17β-oestradiol (nominal concentration: 0.1 µg L−1), but none of the chemical exposures induced intersex. Intersex was revealed in seven out of the nine native populations of C. edule sampled at peak reproductive season. The highest incidence and most severe case of intersex were reported at Lower Anderton on the River Tamer which also had a significantly female-biased sex ratio. Additionally, the dominant trematode family was the Bucephalaidae. Parasitic infection influences the maturity of C. edule by lowering both mean gonad index and condition index. These results suggest that endocrine disrupting chemicals could be contributing factors towards the development of intersex in C. edule.

The synchrony of spawning is of paramount importance to successful coral reproduction. The precise timing of spawning is thought to be controlled by a set of interacting environmental factors, including regional wind field patterns, timing of the sunset, and sea surface temperatures (SST). Climate change is resulting in increased SST, which is causing physiological stress in corals and could also be altering spawning synchrony and timing. In this study, we examined the effect of increasing seawater temperature by 2°C for 1 month prior to the predicted spawning time on reproduction in the coral Acropora digitifera. This short period of elevated temperature caused spawning to advance by 1 day. In animals incubated at elevated temperature, egg number per egg bundle did not change, however, egg volume significantly decreased as did sperm number. Our results indicate that temperature is acting both as a proximate cue to accelerate timing and as a stressor on gametogenesis to reduce fecundity. This finding suggests that increasing SSTs could play a dramatic role in altering reproductive timing and the success of corals in an era of climate change.

Cliona delitrix is one of the most abundant and destructive coral-excavating sponges on Caribbean reefs. However, basic aspects of its reproductive biology, which largely determine the species propagation potential, remain unknown. A 2-year study (October 2009 to September 2011) was conducted to determine the reproductive cycle and gametogenesis of a C. delitrix population located in a shallow reef in Florida, USA. Mesohyl tissue collected from randomly chosen and tagged sponge individuals was sampled one to several times a month, and analysed by light and transmission electron microscopy (TEM). Cliona delitrix is oviparous and gonochoric, except for a few simultaneous hermaphroditic individuals. The C. delitrix reproductive cycle in Florida is from April to December, and is triggered by an increase in seawater temperature to 25°C. Oogenesis and spermatogenesis were asynchronous among individuals; with different cohorts of oocytes co-occurring in females, and spermatic cysts in males. Granulose cells acted as nurse cells, contributing to the growth and maturation of both female and male gametes. Spawning of gametes was not always synchronized with full moon phase. Unlike most other oviparous sponges, the reproductive cycle of C. delitrix is versatile and includes multiple spawning events during the summer of each year. This characteristic maximizes sponge propagation on coral reefs during the warmer months of the year, particularly when thermal stress induces coral mortality. This aspect, combined with its success on polluted areas, make C. delitrix a suitable bioindicator of coral reef health.

The annual gametogenic cycle in the blue mussel, Mytilus edulis L. was investigated from May 2010 to May 2011 in two locations in north-western Iceland. Food availability (chlorophyll-a) was high in early spring at both sites, with a summer maximum in July in Steingrimsfjördur but an autumn maximum in September in Patreksfjördur. Gonad development started in November at both sites, with ripe gametes still visible in the follicles, and continued throughout the year, except for a very short time prior to spawning in Steingrimsfjördur. Gametes developed mostly between November and May. Spawning was protracted, lasting from July to early winter at both sites. The main spawning events occurred from August (11.7°C) to October (9.4°C) in Patreksfjördur, but from July (9.8°C) to October (8.6°C) in Steingrimsfjördur

A one-year study of the reproductive biology of a population of Diopatra marocensis at the Villaviciosa estuary, northern Spain, was undertaken with emphasis on brooding behaviour, larval development and gametogenesis. Field observations together with a histological study of monthly collected individuals revealed that the population was iteroparous, reproducing annually during a short breeding season extending from March to June. The study demonstrated that all individuals of the population at Villaviciosa estuary were producing at the same time eggs and sperm providing for the first time evidence for the occurrence of simultaneous hermaphroditism in D. marocensis. Mature sperm was of the ent-aquasperm type and was stored in modified nephridial chambers in brooding individuals. In the latter, eggs in advanced cleavage phases were also observed inside the coelom suggesting that fertilization was internal. Eggs in late vitellogenesis presented micronucleoli in the periphery of the nucleus, a phenomenon reported previously for few polychaete species. Pure males and females were never found. It is here suggested that in D. marocensis population size may influence sex allocation and that under conditions of low population densities simultaneous hermaphrodites may be favoured. The study of the larval development confirmed the direct development reported previously for populations of Morocco and Portugal. Further results suggest consumption of nurse eggs and adelphophagy by developing larvae.

Testes and ovaries of Enteroctopus megalocyathus collected along the Patagonian Atlantic coast were analysed histologically to validate the macroscopic maturity scales adopted for this species. Changes through the course of development of the seminiferous tubules and of the oocyte/follicular cell complexes were characterized and these were classified into five and six microscopic categories of development respectively. A histological maturity index, based on the frequencies of microscopic categories, was used to assess the correspondence between macroscopic maturation stages and the microscopic level of development of the gonadal tissue. Seminiferous tubules showed a regular and progressive pattern of microscopic development within each macroscopic stage and between consecutive macroscopic stages. However, a minority of males exhibiting seminiferous tubule with sperm did not display macroscopic characteristics of the mature-spawning stage. In females, an overlapping of microscopic categories was observed in maturing macroscopic stages. Previtellogenic oocytes were not present at mature-spawning or spent stages. Significant changes in the histological maturity index were observed between consecutive macroscopic stages, confirming the validity of macroscopic maturity scales of both sexes. In addition, by considering both macroscopic and microscopic criteria, it was possible to determine the overall state of development and functioning of the reproductive system during sexual maturation of this species.

A technique was developed for dissection and isolation of male germ cells in the oyster Crassostrea gigas. This procedure can provide cells for the exploration of processes involved in the reproductive physiology of bivalves. Spermatogonia were chosen because of their essential role in spermatogenesis and the impact of gonia proliferation on reproductive effort. A non lethal method for determining sex and reproductive cycle stage was first validated in oysters. This first step was essential in order to constitute a homogeneous pool of oysters at the same stages of gametogenesis. Germ cell fractions were then obtained from a density gradient, and enrichment of each fraction was ratified by electron microscopy and by means of a 2-parameter flow cytometry procedure (DNA and mitochondrial staining). A significant enrichment in spermatogonia and spermatocytes was confirmed by the increased expression of markers of proliferative cells (proliferative cell nuclear antigen, PCNA) and early germ cells (oyster vasa-like gene). A preliminary cell sorting procedure is also reported, which was applied to fractions enriched in spermatogonia.

We examined the density and reproductive characteristics of the Israeli population of the cryptic species complex Dendropoma petraeum—a Mediterranean endemic reef-building gastropod. The population is gonochoristic, with a peak of gonad activity from October–November to May–June. We present here for the first time histological evidence of its reproductive cycles, hermaphroditism and gonadal parasitic infestation. The characteristics studied (gonad activity period, number of egg capsules per female, capsule size and number of eggs/embryos per capsule) were markedly different from those reported for the D. petraeum-complex population in south-east Spain. Our findings on the population's reproductive characteristics provide new information on the differences between the Mediterranean populations of D. petraeum-complex.

We conducted a histological investigation of the ovaries and testes during the gametogenesis and spawning in the giant jellyfish Nemopilema nomurai, which has bloomed in East Asian marginal seas almost annually since 2002. Oocytes arising from the ovarian epithelium make intimate contact with special epithelial cells, called trophocytes, which have microvilli on the subgenital sinus side, Golgi complexes and vesicles in the cytoplasm. In the early vitellogenic stage, yolk bodies occur in the ooplasm adjacent to the trophocytes, suggesting that the trophocytes transfer nutrients from the subgenital sinus to the oocyte. Later, yolk bodies are formed by the Golgi complexes in the entire ooplasm and accumulate until the oocyte matures. In the late vitellogenic stage, the oocyte separates from the trophocytes and forms microvilli on its surface, indicating nutrient uptake from the surrounding mesoglea. Nutrient support from the trophocytes in the early vitellogenic stage may make the oocytes mature rapidly after medusae are physically damaged. Microvilli-rich epithelial cells also associate with sperm follicles where spermatocytes arise from the follicle cells and accumulate, but their function in nutrient uptake is possibly less than that of trophocytes according to their morphology. During ovulation, which takes 1.5 hours after light exposure, trophocytes separate from each other and make an ovulation pit where the oocyte passes out to the subgenital sinus with the surrounding basal lamina. Spermiation occurs 5–20 minutes after light exposure, and spermatozoa are liberated through the spermiation pit that was formed by which the microvilli-rich cells dissociate. The trophocytes in ovaries and microvilli-rich cells in testes have important roles not only in the gametogenesis but also in the spawning of N. nomurai.