Atresia of ovarian follicles in fishes, and implications and uses in aquaculture and fisheries

Atresia of ovarian follicles, that is the degenerative process of germ cells and their associated somatic cells, is a complex process involving apoptosis, autophagy and heterophagy. Follicular atresia is a normal component of fish oogenesis and it is observed throughout the ovarian cycle, although it is more frequent in regressing ovaries during the postspawning period. An increased occurrence of follicular atresia above physiological rates reduces fish fecundity and even causes reproductive failure in both wild and captive-reared fish stocks, and hence, this phenomenon has a wide range of implications in applied sciences such as fisheries and aquaculture. The present article reviews the available literature on both basic and applied traits of oocyte loss by atresia, including its morpho-physiological aspects and factors that cause a supraphysiological increase of follicular atresia. Finally, the review presents the use of early follicular atresia identification in the selection process of induced spawning in aquaculture and the implications of follicular atresia in fisheries management

Reproductive Biology Of The Shi Drum (Umbrina Cirrosa) In Captivity And Induction Of Spawning Using Gnrha

The reproductive biology of the shi drum (Umbrina cirrosa) in culture was histologically exam- ined and sperm quality was monitored during an entire reproductive period. Already in April, the ovary contained oocytes in all stages of maturation, from primary oocytes to full vitellogenesis, as expected from a group-synchronous multiple-batch spawning fish. Vitellogenesis of the first batch of oocytes occurred very rapidly and their mean diameter (500 μm) did not increase sig- nificantly (p>0.05) as the reproductive period proceeded. The spermiation index peaked in May- June, but fish never produced copious amounts of milt upon abdominal pressure. The sperma- tozoa motility percentage remained unchanged throughout the spawning season (80%) and a significant percentage (40%) maintained viability after overnight storage at 4°C. Sperm density and motility duration increased during the reproductive period and varied 13-26 x109 spermato- zoa/ml and 26-40 s, respectively. Spontaneous spawning was not observed during the two-year study. Injection of post-vitellogenic females with an agonist of gonadotropin-releasing hormone (GnRHa) was successful in inducing a single spawning after two days, with fertilization, hatch- ing and 4-day larval survival rates of 65%, 42-76% and 46-80%, respectively. The results under- line the failure of female shi drum in culture to undergo final oocyte maturation and, although GnRHa injection was effective in inducing spawning of viable eggs, multiple treatments did not induce multiple spawns, as was expected from fish with multiple-batch group-synchronous ovar- ian biology

Gamete quality and management for in vitro fertilisation in meagre (Argyrosomus regius)

The aquaculture of meagre (Argyrosomus regius) requires methods for the control of reproduction that enable the production of families from specific individuals for selective breeding programs. We experimentally determined the parameters required for an in vitro fertilisation protocol. A total of 14 females and 5 males (mean ± S.D. weights of 20.45 ± 6.22 and 15.94 ± 2.75 kg, respectively) were used. Selected females had vitellogenic oocytes >550 μm in diameter and males had fluid sperm upon application of abdominal pressure. Both sexes were treated with an injection of 15 μg kg−1 of gonadotropin-releasing hormone agonist (GnRHa) to induce oocyte maturation/ovulation and enhance sperm production. To determine the timing of ovulation and window of high egg viability, females were stripped serially every 2.5 h beginning 35 h after GnRHa treatment. Sperm was obtained 24 h after GnRHa treatment and was diluted 1/4 in modified Leibovitz for storage at 4 °C until use. Sperm quality parameters such as percentage initial spermatozoa motility, duration of motility, velocity and density were determined using computer assisted sperm analysis (CASA). In vitro inseminations were made in duplicate or triplicate batches of eggs from each spawn by mixing 0.5–1 mL of eggs, 20–40 μL diluted sperm (pooled from two males) and 100 mL of seawater. Fertilisation success was examined at spermatozoa (spz): egg ratios between ~2000 and 400,000 spz egg−1. The optimal time for stripping ovulated females was ≤3 h after ovulation, which was the window of optimal egg viability. Ovulation under the conditions of this study was close to 38 h after GnRHa treatment, with a range from 35 to 41 h. Beginning from 3 h after ovulation, egg viability declined probably due to overripening. Sperm diluted in Leibovitz maintained motility and velocity for as long as 7 h after collection. Spermatozoa motility (%) and average path velocity (VAP, μm/s) of sperm samples obtained from males before GnRHa injection declined rapidly after activation compared to the samples obtained 24 h post-injection, with significant decreases respectively after 75 and 45 s. A minimum ratio of 150,000 spermatozoa egg−1 was necessary to ensure high fertilisation success. The acquired knowledge of the present study will aid the aquaculture industry and future research on selective breeding programs for meagre.info:eu-repo/semantics/acceptedVersio

Induction of ovulation and spawning in the Mediterranean red porgy, Pagrus pagrus, by controlled delivery and acute injection of GnRHa

Gonadotropin-releasing hormone analogue (GnRHa) in the form of saline injections or sustained-release microspheres was used to induce oocyte maturation, ovulation, and spawning in captive red porgy (Pagrus pagrus). Individually tagged vitellogenic females (n = 9 or 10) were treated at the beginning of the spawning season (March) with 20 μg/kg body weight (bw) GnRHa-loaded microspheres, a single injection of 20 μg/kg bw dissolved in saline, or physiological saline (control). Females were placed in tanks (one tank per treatment) connected to overflow egg collectors and monitored for 11 days. In addition to the eggs collected from the tank overflow, eggs were stripped from the fish on a daily basis. Only one spawn was obtained from the control fish, probably from a single female, given the small relative fecundity (700 eggs/kg bw). On the contrary, treatment with a GnRHa injection produced two spawns (9 and 11 days after treatment) and 50% of the fish ovulated. Treatment with GnRHa microspheres induced seven spawns (3 and 6-11 days after treatment) and 100% of the females ovulated. Females did not spawn all the eggs ovulated on a particular day, evident from the significant number of eggs obtained by manual stripping. Egg quality did not significantly differ among treatments, whereas number of spawned eggs and total relative fecundity were significantly higher in fish treated with GnRHa microspheres (ANOVA, p<0.05). The results demonstrate the potential of GnRHaloaded microspheres to induce spawning in red porgy, as a method of overcoming spawning failures in commercial hatcheries

Developmental expression of DAX1 in the European sea bass, Dicentrarchus labrax: lack of evidence for sexual dimorphism during sex differentiation

DAX1 (NR0B1), a member of the nuclear receptors super family, has been shown to be involved in the genetic sex determination and in gonadal differentiation in several vertebrate species. In the aquaculture fish European sea bass, Dicentrarchus labrax, and in the generality of fish species, the mechanisms of sex determination and differentiation have not been elucidated. The present study aimed at characterizing the European DAX1 gene and its developmental expression at the mRNA level

Spawning induction of first-generation (F1) greater amberjack Seriola dumerili in the Canary Islands, Spain using GnRHa delivery systems

The development of a sustainable aquaculture industry requires the closing of the life-cycle of a potential species in captivity, and the establishment of breeding selection programs using hatchery-produced breeding stocks. The greater amberjack Seriola dumerili is a cosmopolitan pelagic species that has been considered as a good candidate for the species diversification of aquaculture production in the Mediterranean region. However, commercial production is still very limited due to bottlenecks in reproduction, larval rearing and management control during grow out. The aim of the present study was to examine the reproductive development of hatchery-produced greater amberjack and to develop a spawning induction protocol based on the use of gonadotropin releasing hormone agonist (GnRHa) in a controlled-release delivery system. The results showed that hatchery-produced greater amberjack undergo normal gametogenesis and can be induced to undergo maturation, ovulation and spawning after multiple administrations of GnRHa implants, over an extended spawning period lasting from May to September in the Canary Islands, Spain. The use of GnRHa-delivery systems resulted in multiple spawns of fertilized and viable eggs. Egg production was high and egg quality adequate for the implementation of larval rearing for commercial purposes. The handling required to administer the GnRHa implants during the prolonged spawning season did not result in any negative effect on the welfare and reproductive performance of the fish based on evaluation of several biochemical parameters. The developed reproduction control method shows great potential to advance the commercial production of greater amberjack, by enabling the use of hatchery-produced broodstocks for further breeding selection.Postprin

Spontaneous spawning of Atlantic bluefin tuna Thunnus thynnus kept in captivity

Beginning 48-72 h after GnRHa treatment in 2009, massive spawning occurred for 17 days, with a daily maximum of 34 million eggs collected (SELFDOTT, 2010). On 17th June 2010 spontaneous spawning started in cage R2. From this day onwards and more or less daily viable eggs were obtained from both cages so no hormonal implants were used. Fish spawned intermittently for a period of 34 days in both cages, with a total of 58 million eggs collected. Cage R1 spawned for 26 days and produced 48 million eggs and cage R2 spawned for 28 days and produced 10 million eggs (Figure 1). As happened in 2009 (SELFDOTT, 2010), due to the almost non-existence of currents throughout the spawning period, almost all the eggs were collected at the surface, held by the plastic curtain. Only a small amount of eggs were captured by the egg collectors, which were designed to operate under current. Eggs were transported to the IEO facilities in Mazarrón (SE- Spain) where they were incubated. On average, the hatching rate was above 90%. No difference in hatching success was observed between 2009 (from hormonally treated cage) and 2010 (from spontaneously spawning fish).Commission of the European Communities, specific RTD program of Framework Program 7, SELFDOTT, “From capture based to self-sustained aquaculture and domestication of bluefin tuna, Thunnus thynnus” (GA 212797)

Male germ cell proliferation and apoptosis in sexually immature meagre Argyrosomus regius (Asso, 1801) treated with recombinant follicle stimulating hormone

The meagre Argyrosomus regius (Asso, 1801) is a marine fish species that has an increasing aquaculture production in Europe. Lowering the age at maturity of hatchery-produced juveniles would support meagre aquaculture by reducing time between generations in selective breeding programs and reducing industrial costs for broodstock maintenance. The aim of this work was to assess the effects of a treatment with recombinant follicle stimulating hormone (rFsh), produced in ovarian cells of Chinese hamsters, on male germ cell proliferation and apoptosis in sexually immature meagre. The rFsh-treated fish had higher gonadosomatic index, larger seminiferous tubules, more abundant luminal spermatozoa, a lower density of anti-PCNA positive single A spermatogonia, a higher density of anti-PCNA positive spermatocysts and a lower incidence of germ cell apoptosis than control groups. The present study demonstrated the effectiveness of the produced rFsh in stimulating testis development and spermatogenesis in pre-pubertal meagre. Moreover, the rFsh treatment proved to be highly efficient in removing the apoptotic block of spermatogenesis observed in juvenile meagre, allowing spermatogonial survival and progress towards meiosis. The administration of rFsh did not stimulate spermatogonial self-renewal, a process whose control still needs to be elucidated.info:eu-repo/semantics/publishedVersio

Spawning kinetics and parentage contribution of European sea bass (Dicentrarchus labrax) broodstocks, and influence of GnRHa-induced spawning

Abstract Increasing parentage contribution in aquaculture broodstocks is important, in order to take full advantage of the available genetic makeup of the chosen fish, and to avoid inbreeding and loss of allele diversity over subsequent production generations. European sea bass (Dicentrarchus labrax) broodstocks were evaluated over two reproductive seasons to examine spawning kinetics, egg production, and parentage contribution during spontaneous/volitional spawning. In addition, we obtained preliminary results on the potential of a hormonal therapy to synchronize spawning and increase parentage contribution. Spawning lasted between 25 and 66 days in January-March and consisted of 12–21 daily spawns per broodstock, with individual females spawning 1–5 times and males participating in 1–8 spawns during each reproductive season. Daily fecundity was variable during the season, without any trend, and so were all the examined egg/larval quality parameters. Parentage assignment of the produced families indicated that the majority of progeny from a whole season may belong to a very small number of breeders, with four females producing up to 80 % of the analyzed eggs, while a single male may sire up to 57 % of the fertilized eggs. No significant improvement in the overall parentage contribution was obtained with the hormonal treatment, using gonadotropin releasing hormone agonist (GnRHa). Nevertheless, the daily fecundity was higher, and parentage of the eggs from the first spawn after GnRHa treatment was more equally distributed to multiple males/females, compared to any volitional spawns. The study demonstrates the need to further improve parentage contribution in European sea bass aquaculture, through synchronization of maturation and spawning. Although the GnRHa induction experiment was not replicated in the present preliminary study, the results suggest that hormonally-induced synchronization of maturation may have the potential of producing a larger number of progenies from more families, from where to select the next generation of breeders for a breeding program

Endocrine regulation of long-term enhancement of spermiation in meagre (Argyrosomus regius) with GnRHa controlled-delivery systems.

Abstract Meagre (Argyrosomus regius) undergo spermatogenesis and spermiation when reared in captivity, but often produce low milt volumes, sometimes with reduced quality and for a limited time period. In the present study we a) compared the efficiency of gonadotropin releasing hormone agonist (GnRHa) implants versus injections on testicular stimulation and spermiation enhancement, b) investigated the effect of GnRHa on the endocrine spermiation regulation (sex steroid hormones), and c) evaluated a commercial induced spawning simulation scenario. Firstly, males (n = 5) were injected with 15.0 ± 0.2 μg GnRHa kg−1 (Injections) or implanted with 51.0 ± 5.1 μg GnRHa kg−1 (Implant) and compared their sperm production response. Secondly, the best hormonal treatment (Implant) was tested treating males (n = 8) with 57.5 ± 7.5 μg GnRHa kg−1 every 3 weeks for a period of 70 days. Milt production was improved by the GnRHa implants with only minor sperm quality alterations (improved sperm motility percentage). Elevated plasma testosterone (T) and 11-ketotestosterone (11-KT) levels were recorded in response to GnRHa implants, while no significant difference for 17,20β-dihydroxy-4-pregnen-3-one (17,20β-P) was observed. In the commercial induced spawning simulation, it was shown that meagre females are capable of on-demand induction of spawning at random intervals (5–21 days) using GnRHa injections, over a period of at least 2.5 months. During this period, spermiation enhancement was achieved with GnRHa implants every 3 weeks, producing sperm with stable, in general, quality and motility parameters. Percentage of motile cells, motility duration and density fluctuated significantly, but remained within levels that are considered appropriate for high fertilization success in this species