A. Corriero

Life History and Stock Structure of Atlantic Bluefin Tuna (Thunnus thynnus)

Our understanding of the biology of Atlantic bluefin tuna (Thunnus thynnus) has increased profoundly in the last decade, and the progress is attributed to the development and application of a variety of novel tools. Here we provide a comprehensive examination of available data on the life history and stock structure of T. thynnus by re-examining current databases and literature and highlighting findings from recent studies using approaches such as archival tags and natural markers (e.g., genetics, otolith chemistry). The present review provides a detailed synthesis on the reproductive biology, feeding ecology, growth, mortality, migration, and stock structure of T. thynnus. In addition to characterizing key life history attributes and discussing stock-specific (east versus west) differences, the implication of trans-Atlantic movement and mixing are addressed. We also identify significant data needs that still exist and must be addressed to promote effective management and rapid recovery of T. thynnus populations.

Preparation and Administration of Gonadotropin-Releasing Hormone Agonist (GnRHa) Implants for the Artificial Control of Reproductive Maturation in Captive-Reared Atlantic Bluefin Tuna (Thunnus thynnus thynnus)

Abstract Mature migrating Atlantic bluefin tuna (Thunnus thynnus thynnus) were captured in the Mediterranean Sea with a purse seine and reared in floating cages for 2 to 3 years. During the natural spawning period (June–July) of two consecutive years, fish were randomly implanted underwater with a controlled-release delivery system (implant) loaded with gonadotropin-releasing hormone agonist (GnRHa), in order to induce final oocyte maturation (FOM), ovulation/spermiation, and spawning. At the time of sampling, males were significantly larger than females (ANOVA, P < 0.001), having a mean (± SE) fork length and body weight of 190 ± 3 cm and 122 ± 5 kg, compared to 176 ± 3 cm and 94 ± 4 kg of females, respectively. All fish were reproductively mature, with their age ranging between 5 and 12 years and males being a year older, on average. After GnRHa implantation, fish were monitored for spawning and the release of eggs, and were sacrificed at different times after hormone treatment in order to examine the progressive effect of the treatment on gonad maturation. The in vitro GnRHa release from the produced implants was maximal during the first 2 d, with a mean (± SE) release of 525 ± 166 μ g GnRHa implant−1 day−1. The plasma GnRHa profile in vivo reflected the release in vitro, and statistically significant elevations in plasma GnRHa levels were measured until 7 d after treatment (ANOVA, P < 0.01). The underwater implantation procedure was improved between 2004 and 2005, requiring an average (± SD) of 3.1 ± 1.4 min for each fish, and was 64 and 84% successful in 2004 and 2005, respectively. There were no differences between the histological appearance of the testes of GnRHa-treated and control males, and almost all of them contained intra-testicular spermatozoa. However, the proportion of spermiating control males (n = 17) was only 12% compared to 26% for the GnRHa-implanted males (n = 19). Also, there were no differences between controls and GnRHa-implanted fish in sperm concentration, initial spermatozoa motility, or duration of forward motility, which ranged between 29.02–48.54 × 1010 spermatozoa ml−1, 58–63% and 8–9 min, respectively. Final oocyte maturation (FOM) and post-ovulatory follicles (POFs) occurred in 63% and 88%, respectively, of the GnRHa implanted females (n = 16), compared to 0% and 21%, respectively, of the control females (n = 14). In addition, two GnRHa-implanted females in 2005 were found to be ovulated at the time of sacrifice, and their eggs were fertilized in vitro with sperm from spermiating males, which resulted in viable embryos and larvae. Finally, although spawning was not observed, fertilized eggs were collected from the cages. Larvae produced from these eggs were identified as Atlantic bluefin tuna, demonstrating that the present GnRHa implantation method can be used to induce FOM, ovulation/spermiation, and spawning in captive-reared Atlantic bluefin tuna.

Atlantic Bluefin Tuna (Thunnus Thynnus) Farming and Fattening in the Mediterranean Sea

The Atlantic bluefin tuna (Thunnus thynnus) is one of the tunas with the highest commercial value and it is supporting the capture-based tuna aquaculture industry in the Mediterranean Sea. This is a seasonal activity and it involves the capture of fish from the wild and their rearing in sea cages for periods ranging between 3 months to 2 years. Short-term rearing is done mainly to: (a) achieve a greater body fat percentage and (b) obtain a better price by not flooding the market in the brief fishing period. Due to the increasing fear of a collapse of the fishery, the International Commission for the Conservation of Atlantic Tunas currently reduced the total allowable catches for 2010 to 13,500 mtn from 32,000 mtn previously. Therefore, there is great interest in establishing a proper and sustainable tuna aquaculture industry. This necessitates the development of specific technologies for tuna aquaculture that will not rely on captured individuals from the wild, as it is practiced today. This article reviews the methods used for the farming and fattening of the species in the Mediterranean Sea, and the current status of the efforts at controlling reproduction in captivity.

Evidence of a high percentage of intersex in the Mediterranean swordfish (Xiphias gladius L.)

The first evidence of the presence of intersexuality in a wild population of Mediterranean swordfish (Xiphias gladius L.) is reported. Forty of 162 specimens (25%) macroscopically classified as males, showed the presence of female germ cells within the testes. In two specimens grouped previtellogenic oocytes were present; all the other specimens possessed single scattered previtellogenic oocytes. The presence of vitellogenin was demonstrated immunohistochemically in the liver of both intersex and normal males. These findings could be due to the exposure to oestrogen-mimicking substances.

Incidental captures of sea turtles by swordfish and albacore longlines in the Ionian sea

Incidental catches in pelagic longline fishing pose a serious threat to sea turtle populations throughout the world’s seas and oceans. In this work, carried out in the framework of the EC-DG-Fisheries 98/008 project, information on sea turtle catch rates from swordfish and albacore longline fishing activities observed in Italian waters off the Ionian Sea during 1999 and 2000, are reported. In addition, biometric data, health status, and tagging return rate of sea turtles captured are provided. A total of 200 sea turtles were caught (198 loggerhead turtles and 2 green turtles), comprising 0.5–15.7% of the total catch in number of individuals. The estimates of the sea turtles caught by the total fishing effort of both longlines in the whole study area were 1084 specimens in 1999 (95% Cl=667–1502) and 4447 specimens in 2000 (95% Cl=3189–5705). Although all sea turtles were released alive, nearly half of them had hooks that could not be removed and remained deeply embedded in the digestive tract.

GnRHa-mediated stimulation of the reproductive endocrine axis in captive Atlantic bluefin tuna, Thunnus thynnus.

A controlled-release implant loaded with GnRH agonist (GnRHa) was used to induce spawning in Atlantic bluefin tuna (Thunnus thynnus) during two consecutive reproductive seasons. The fish were implanted underwater and sampled between days 2 and 8 after treatment. At the time of GnRHa treatment, females were in full vitellogenesis and males in spermiation. There was a rapid burst of pituitary luteinizing hormone (LH) release at day 2 after treatment in GnRHa-treated fish, and circulating LH remained elevated up to day 8 after treatment. In contrast, control fish had significantly lower levels in the plasma, but higher LH content in the pituitary, as observed in many other cultured fishes that fail to undergo oocyte maturation, ovulation and spawning unless induced by an exogenous GnRHa. Plasma testosterone (T) and 17β-estradiol (E(2)) were elevated in response to the GnRHa treatment in females, while 11-ketotestosterone (11-KT) but not T was elevated in males. Even though oocyte maturation and ovulation did occur in GnRHa-induced fish, no significant elevations in 17,20β-dihydroxy-4-pregnen-3-one (17,20β-P) or 17,20β,21-trihydroxy-4-pregnen-3-one (20β-S), in either the free, conjugated or 5β-reduced,3α-hydroxylated forms was observed in fish sampled within 6 days after treatment. Interestingly, a significant peak in plasma free 17,20β-P levels occurred in both males and females at day 8 after treatment. Histological sections of the ovaries in these females contained oocytes at the migrating germinal vesicle stage, suggesting the role of this hormone as a maturation-inducing steroid in Atlantic bluefin tuna. In conclusion, the GnRHa implants activated effectively the reproductive endocrine axis in captive Atlantic bluefin tuna broodstocks, through stimulation of sustained elevations in plasma LH, which in turn evoked the synthesis and secretion of the relevant sex steroids leading to gamete maturation and release.

Comparative study of liver vitellogenin gene expression and oocyte yolk accumulation in wild and captive Atlantic bluefin tuna (Thunnus thynnus L.).

The sequence of vitellogenin A (VgA) and vitellogenin B (VgB) cDNAs in Atlantic bluefin tuna (Thunnus thynnus L.) were determined, and vitellogenin expression levels in the liver and oocyte yolk accumulation were compared in wild and captive-reared individuals. Liver and ovary samples were taken from 31 individuals reared experimentally in three commercial Atlantic bluefin tuna fattening sites in the Mediterranean Sea and from 33 wild individuals caught by commercial traps during the fishs migration towards their Mediterranean spawning grounds. The total length of VgA cDNA was 5585 nucleotides and that of VgB was 5267 nucleotides. The identity and similarity between deduced amino acid sequences of VgA and VgB were 60% and 78%, respectively. The Atlantic bluefin tuna VgA and VgB amino acid sequences have high similarities with those of other teleost fishes. Relative levels of VgA and VgB mRNAs were low in April, increased significantly during the reproductive period in May and June, and declined in July. There was a trend towards higher relative levels of VgA and VgB mRNAs in captive fish compared to wild individuals during the reproductive period. The surface occupied by eosinophilic yolk granules in fully vitellogenic oocytes, as well as the frequency of oocytes in late vitellogenesis, was significantly higher in captive compared to wild individuals. The study suggests that the experimental conditions under which Atlantic bluefin tuna individuals were reared allowed the occurrence of normal vitellogenesis, based on gene expression of VgA and VgB in the liver and yolk accumulation in the oocytes. The higher yolk accumulation and frequency of vitellogenic oocytes observed in the ovaries of captive fish suggest that improvements in feeding practices may result in an improved vitellogenic process.

Histological and immunohistochemical investigation on ovarian development and plasma estradiol levels in the swordfish (Xiphias gladius L.)

The paper reports a histological and immunohistochemical description of oocyte growth and ultrastructural aspects of zona radiata (ZR) formation as well as the relationship between plasma estradiol-17beta, (E2) levels and ovarian development in swordfish (Xiphias gladius L.) from the Mediterranean Sea. Ovaries were inactive during March to mid April; maturation occurred during late April to June and spawning in June and July. Zona radiata formation starts, as Pas positive material, in oocytes at the lipid stage. In this stage a deposit of electrondense material between oolemma and follicular cells appears. In the cortical alveoli stage and through the early vitellogenic stage, the deposition of a moderately electrondense material occurred on the inner side of the ZR. Finally, in late vitellogenic oocytes a third layer, made of microfibrillar material, appeared. The immunohistochemical analyses revealed that the initial internalisation of hepatic zona radiata proteins (Zrp) in the swordfish oocyte starts before the uptake of vitellogenin (Vtg) and that it is associated with the low previtellogenic E2 plasma levels, while a significant E2 increase in plasma is associated with the beginning of Vtg uptake. This would appear to confirm the hypothesis that the differential and sequential induction of zonagenesis and vitellogenesis may reflect a general feature of teleost oogenesis.

Distribution of sialoglycoconjugates in the oviductal isthmus of the horse during anoestrus, oestrus and pregnancy: a lectin histochemistry study

The distribution of sialic acid residues as well as other glycosidic sugars has been investigated in the horse oviductal isthmus during anoestrus, oestrus and pregnancy by means of lectin and pre-lectin methods. Ciliated cells and non-ciliated (secretory) cells exhibited different lectin binding profiles that were found to change during the investigated stages. Ciliated cells did not show any reactivity in the basal cytoplasm, while the supra-nuclear cytoplasm displayed a few of oligosaccharides with terminal and internal alphamannose (Man) and/or alphaglucose (Glc) during oestrus and pregnancy and a moderate presence of oligosaccharides terminating in alphafucose (Fuc) during oestrus; cilia exhibited a more complex glycoconjugate pattern for the presence of oligosaccharides terminating in N-acetylgalactosamine (GalNAc), GalNAcalpha1,3 GalNAcalpha1,3galactose(Gal)beta1,4Galbeta1,4N-acetylglucosamine(GlcNAc), Fuc, sialic acid (Neu5Ac)-aGalNAc belonging or not to the GalNAca1,3GalNAca1,3 Galb1,4 Galb1, 4GlcNAc sequence, and. alphaGalNAc and Neu5Aca 2,6Gal/GalNAc increased during oestrus. Cilia displayed terminal Galbeta1,3 GalNAc in pregnancy, terminal alphaGal in anoestrus and pregnancy and terminal or internal D-GlcNAc during anoestrus and pregnancy, respectively. The whole cytoplasm of non-ciliated cells showed oligosaccharides terminating with alphaGalNAc, Neu5Aca2,6Gal/GalNAc, Neu5Ac GalNAca 1,3GalNAcalpha1,3Galbeta1,4Galbeta1,4GlcNAc during the investigated stages, as well as GlcNAc in anoestrus and pregnancy. The supra-nuclear zone of non-ciliated cells exhibited oligosaccharides with terminal Galbeta1,4GlcNAc and internal Man during oestrus and pregnancy as well as terminal alphaGal and Fuc in oestrus and Neu5Ac-Galbeta1,3GalNAc in pregnancy. The luminal surface of non-ciliated cells showed glycans terminating with alphaGalNAc and/or Neu5Ac GalNAcalpha1,3 GalNAcalpha1,3Galbeta1,4Galbeta1,4GlcNAc in all specimens, oligosaccharides with terminal Galbeta1,4GlcNAc and internal Man during oestrus and pregnancy, Neu5Ac alpha2,6Gal/GalNAc in anoestrus and oestrus, and glycans terminating with Galbeta1,3GalNAc, Neu5A acalpha2,3 Galbeta1, 4GlcNac, Neu5ac-Galbeta1,3GalNAc, Neu5Ac-Galbeta1,4 GlcNAc in pregnancy. These findings show the presence of sialoglycoconjugates in the oviductal isthmus of the mare as well as the existence of great modifications in the glycoconjugates linked to different physiological conditions.

Expression of vitellogenin receptor gene in the ovary of wild and captive Atlantic bluefin tuna (Thunnus thynnus)

The cDNA sequences of vitellogenin receptor proteins (VgR(+) and VgR(-)), containing or lacking the O-linked sugar domain, were determined in Atlantic bluefin tuna (Thunnus thynnus L.). VgR(-) gene expression in the ovary was compared in captive-reared and wild Atlantic bluefin tuna during the reproductive cycle. Gonad samples from adult fish were sampled from 2008 to 2010 from stocks reared in captivity at different commercial fattening operations in the Mediterranean Sea and from wild individuals caught either by traditional tuna traps during their migration towards the spawning grounds in the Mediterranean Sea or by the long-line artisanal fishery. In addition, juvenile male and female Atlantic bluefin tuna were sampled from a farming facility, to obtain baseline information and pre-adulthood amounts of VgR(-). The total length of VgR(+) cDNA was 4006 nucleotides (nt) and that of VgR(-) was 3946 nt. Relative amounts of VgR(-) were greater in juvenile females and in those adults having only previtellogenic oocytes (119 ± 55 and 146 ± 26 folds more than juvenile males, respectively). Amounts of VgR(-) were less in individuals with yolked oocytes (ripening stage, May-June) and increased after spawning in July (92 ± 20 and 113 ± 13 folds more than juvenile males in ripening and post-spawning fish, respectively). These data suggest that regulation of VgR(-) is not under oestrogen control. During the ripening period, greater VgR(-) gene expression was observed in wild fish than in fish reared in captivity, possibly because of (a) differences in water temperature exposure and/or energy storage, and/or (b) an inadequate diet in reared Atlantic bluefin tuna.