Atlantic bluefin tuna tagged off Norway show extensive annual migrations, high site-fidelity and dynamic behaviour in the Atlantic Ocean and Mediterranean Sea.

Atlantic bluefin tuna (ABFT; Thunnus thynnus) is a highly migratory species. To investigate the migrations and vertical behaviours of ABFT migrating to Nordic waters, we deployed pop-up satellite archival transmitting tags on 25 ABFT off Norway (curved fork length: 228-292 cm). We obtained 16 full-year migrations, which differed between individuals, and physically recovered 13 tags, which provided 4699 days of archival depth and temperature data. ABFT occupied waters from the Arctic Circle to as far south as Cabo Verde, Africa, and occupied depths down to 1190 m and temperatures from 0.5 to 27.8°C. During their annual migrations, ABFT spent, on average, 68 days in Norwegian waters, 65 days in the Newfoundland Basin, 35 days around the Canary Islands and 33 days in the West European Basin. Most ABFT entered the Mediterranean Sea with a mean entry date of 13 May and visited known spawning grounds, staying, on average, 44 days. All ABFT with full-year deployments returned to Norwegian waters. ABFT displayed high site-fidelity and dynamic vertical diving behaviours that varied between hotspots and seasons. These spatiotemporal data provide important ecological knowledge for sustainable management and the conservation of the recently recovered eastern ABFT stock.

Unidirectional trans-Atlantic gene flow and a mixed spawning area shape the genetic connectivity of Atlantic bluefin tuna.

The commercially important Atlantic bluefin tuna (Thunnus thynnus), a large migratory fish, has experienced notable recovery aided by accurate resource assessment and effective fisheries management efforts. Traditionally, this species has been perceived as consisting of eastern and western populations, spawning respectively in the Mediterranean Sea and the Gulf of Mexico, with mixing occurring throughout the Atlantic. However, recent studies have challenged this assumption by revealing weak genetic differentiation and identifying a previously unknown spawning ground in the Slope Sea used by Atlantic bluefin tuna of uncertain origin. To further understand the current and past population structure and connectivity of Atlantic bluefin tuna, we have assembled a unique dataset including thousands of genome-wide single-nucleotide polymorphisms (SNPs) from 500 larvae, young of the year and spawning adult samples covering the three spawning grounds and including individuals of other Thunnus species. Our analyses support two weakly differentiated but demographically connected ancestral populations that interbreed in the Slope Sea. Moreover, we also identified signatures of introgression from albacore (Thunnus alalunga) into the Atlantic bluefin tuna genome, exhibiting varied frequencies across spawning areas, indicating strong gene flow from the Mediterranean Sea towards the Slope Sea. We hypothesize that the observed genetic differentiation may be attributed to increased gene flow caused by a recent intensification of westward migration by the eastern population, which could have implications for the genetic diversity and conservation of western populations. Future conservation efforts should consider these findings to address potential genetic homogenization in the species.

A novel protocol for rapid deployment of heart rate data storage tags in Atlantic bluefin tuna Thunnus thynnus reveals cardiac responses to temperature and feeding.

The Atlantic bluefin tuna (ABFT) is a highly prized species of large pelagic fish. Studies of their environmental physiology may improve understanding and management of their populations, but this is difficult for mature adults because of their large size. Biologging of heart rate holds promise in investigating physiological responses to environmental conditions in free-swimming fishes but it is very challenging to anesthetize large ABFT for invasive surgery to place a tag in the body cavity near to the heart. We describe a novel method for rapid deployment of a commercially available heart-rate tag on ABFT, using an atraumatic trocar to implant it in the musculature associated with the cleithrum. We performed three sequential experiments to show that the tagging method (1) is consistently repeatable and reliable, (2) can be used successfully on commercial fishing boats and does not seem to affect fish survival, and (3) is effective for long-term deployments. In experiment 3, a tag logged heart rate over 80 days on a 60-kg ABFT held in a farm cage. The logged data showed that heart rate was sensitive to prevailing seasonal temperature and feeding events. At low temperatures, there were clear responses to feeding but these all disappeared above a threshold temperature of 25.5°C. Overall, the results show that our method is simple, rapid, and repeatable, and can be used for long-term experiments to investigate physiological responses by large ABFT to environmental conditions.

An autofluorescence-based survey of late follicular atresia in ovaries of a teleost fish (Thunnus thynnus).

In this study, the authors examined late atretic follicles in the ovaries of Atlantic bluefin tuna, Thunnus thynnus (Linnaeus 1758), at different times of the year using transmitted light and epifluorescence microscopy. Atresia (degeneration and resorption of developing ovarian follicles) is a natural process involved in fecundity downregulation in teleosts and is substantially enhanced in stressful conditions. Early (α and ß) atretic stages of yolked oocytes have a relatively short duration in seasonally reproducing species, whereas later (γ and δ) atretic follicles (LAF) persist for longer time in the ovary, serving as a sign of previous vitellogenic activity. LAF can thus be used as reliable markers of maturity during non-reproductive periods. Lipofuscin granules accumulate in the cytoplasm of LAF cells as a result of lysosomal digestion of oocyte components. Taking advantage of the well-known autofluorescent properties of lipofuscins, LAF may be identified in unstained histological sections under fluorescence microscopy using appropriate excitation and emission wavelengths. The authors explore in this study the applicability of fluorescence microscopy to provide a fast and effective method to assess late atresia in fishes. This method may be particularly useful to determine sexual maturity in individuals sampled long after the spawning season, where LAF are difficult to detect in standard histological sections. Furthermore, LAF autofluorescence provides a rapid way to quantify late atresia in fishes using image analysis.

In vivo and in vitro assessment of host-parasite interaction between the parasitic copepod Brachiella thynni (Copepoda; Lernaeopodidae) and the farmed Atlantic bluefin tuna (Thunnus thynnus).

The Atlantic bluefin tuna (ABFT; Thunnus thynnus) today represents one of the economically most important species for Croatian fisheries industry. Although the most diverse and abundant parasitofauna is usually found in the largest specimens of wild ABFT, the opposite was observed in captivity where parasite populations significantly decline by the end of the farming cycle. Copepod Brachiella thynni, is a skin parasite frequently parasitizing tuna, whose population also decreases in number throughout the rearing process. In order to better understand the immunity mechanisms underlying ABFT reaction to B. thynni infection, we studied expression profiles of immunity related genes; interleukin 1ß (il1ß), tumour necrosis factors (tnfα1, tnfα2), complement component 4 (c4) and caspase 3 (casp3), in peripheral blood leukocytes (PBLs) during in vitro stimulation by B. thynni protein extracts (i.e. antigens) and in infected tissues at B. thynni parasitation site. Finally, a histopathological analysis of semi-thin and ultra-thin sections of tissues surrounding B. thynii attachment site was performed to evaluate the severity of parasite-induced lesions and identify involved cell lineages. In vitro stimulation of ABFT PBLs with B. thynii antigens caused a dose-depended upregulation of selected genes, among which tnfα1 showed the highest induction by both concentrations of B. thynni protein extract. However, targeted genes were not significantly upregulated in the infected tissue. Also, no significant alterations in ultrastructure of epithelial layers surrounding B. thynii attachment site were noticed, except local tissue erosion, necrosis of squamous epithelium and proliferation of rodlet and goblet cells. Our results suggest that B. thynii has evolved strategies to successfully bypass both innate immune response and the connective-tissue proliferation processes. Therefore, the observed disappearance of this copepod by the end of the rearing process is more likely related to its limited lifespan on the host and its inability to complete the life cycle in the rearing cages, rather than host's reaction.

Prevalence, intensity and pathology of the nasal parasite Nasicola hogansi in Atlantic bluefin tuna (Thunnus thynnus).

Ectoparasitic flatworms of Nasicola (Monogenoidea: Capsalidae), which infect nasal epithelium of true tunas (Thunnus spp.), are not well studied, nor have their impacts on the host's olfactory organ been evaluated. Infections of Nasicola hogansi on Atlantic bluefin tuna, Thunnus thynnus, were investigated with emphasis on the relationship between infection prevalence, abundance and mean intensity with bluefin tuna size, sex, body condition and capture month, as well as histopathological effects. Commercially caught Atlantic bluefin tuna (n = 161, 185-305 cm curved fork length) from the Gulf of Maine were sampled during June through August 2009 for infections by N. hogansi. A total of 247 specimens of N. hogansi were collected, with a prevalence of 45.3%, mean abundance of 1.57 (CI: 1.21-2.03) and mean intensity of 3.45 (CI: 2.91-4.22). Neither fish sex nor landing month had a significant effect on parasite parameters. Larger and better-conditioned Atlantic bluefin tuna had a higher mean intensity of infection. Pathology associated with infection by N. hogansi included extensive necrosis, sloughing of the nasal epithelium and associated inflammation of underlying connective tissues. Further epidemiological and pathological study of this host-parasite system is warranted since impaired olfaction, if present, could adversely affect spawning and migration of this top ocean predator.

First estimates of metabolic rate in Atlantic bluefin tuna larvae.

Atlantic bluefin tuna is an iconic scombrid species with a high commercial and ecological value. Despite their importance, many physiological aspects, especially during the larval stages, are still unknown. Metabolic rates are one of the understudied aspects in scombrid larvae, likely due to challenges associated to larval handling before and during respirometry trials. Gaining reliable estimates of metabolic rates is essential to understand how larvae balance their high growth needs and activity and other physiological functions, which can be very useful for fisheries ecology and aquaculture. This is the first study to (a) estimate the relationship between routine metabolic rate (RMR) and larval dry weight (DW) (mass scaling exponent) at a constant temperature of 26°C, (b) measure the RMR under light and darkness and (c) test whether the interindividual differences in the RMR are related to larval nutritional status (RNA/DNA and DNA/DW). The RMR scaled nearly isometrically with body size (b = 0.99, 0.60-31.56 mg DW) in contrast to the allometric relationship observed in most fish larvae (average b = 0.87). The results show no significant differences in larval RMR under light and darkness, suggesting similar larval activity levels in both conditions. The size explained most of the variability in RMR (97%), and nutritional condition was unrelated to the interindividual differences in routine metabolism. This is the first study to report the metabolic rates of Atlantic bluefin tuna larvae and discuss the challenges of performing bioenergetic studies with early life stages of scombrids.

Determination of Hg in Farmed and Wild Atlantic Bluefin Tuna (Thunnus thynnus L.) Muscle.

Mercury (Hg) is a well-known toxic element, diffused in the environment, especially in the Mediterranean Sea which is rich in cinnabar deposits. Mercury bioaccumulation in fish is of great concern, especially for top-level aquatic predators (e.g., shark, tuna, swordfish) and above all for species of large human consumption and high nutritional value. This work aimed to determine Hg concentrations in farmed and wild Atlantic Bluefin tuna (Thunnus thynnus) caught in the Mediterranean area in order to evaluate the level of Hg bioaccumulation. selenium (Se) content was also determined, since this element is an antagonist of mercury toxicity. Mercury and Se were analysed by atomic absorption spectrometry after microwave digestion of the samples. Hg content in farmed tuna was below the legal limit (1 mg/kg, wet weight, w.w.) for all specimens (0.6 ± 0.2 mg/kg), whereas the wild ones had a content over the limit (1.7 ± 0.6 mg/kg); Se concentration was higher in farmed specimens (1.1 ± 0.9 mg/kg) compared to wild ones (0.6 ± 0.3 mg/kg). A safe seafood could show a Se/Hg ratio >1 and a health benefit value (HBVSe) > 0: farmed tuna had higher values than the wild specimens (Se/Hg 5.48 vs. 1.32; HBVSe 11.16 vs. 0.29). These results demonstrate that for Hg, there is a better risk/benefit ratio in farmed T. thynnus. making it safer than wild tuna.

Performance, feed utilization, and hepatic metabolic response of weaned juvenile Atlantic bluefin tuna (Thunnus thynnus L.): effects of dietary lipid level and source.

Two trials were performed using extruded diets as on-growing feeds for weaned Atlantic bluefin tuna (Thunnus thynnus; ABT) to establish adequate dietary levels of both lipid and omega-3 long-chain polyunsaturated fatty acids (LC-PUFAs), and impacts on lipid metabolism via liver gene expression. In trial A, ABT were fed with either a commercial feed (Magokoro®; MGK) as a reference diet or two experimental feeds differing in lipid levels (15 or 20%) using krill oil (KO) as the single lipid source in order to estimate suitable lipid content. Fish fed MGK displayed the highest growth, followed by 15KO, and therefore a dietary lipid content of 15% was considered preferable to 20% at this stage. In trial B, fish were fed MGK, 15KO, or a feed containing 15% lipid with a blend of KO and rapeseed oil (RO) (1:1, v/v; 15KORO). Fish fed 15KO and 15KORO showed no difference in weight gain, specific growth rate, and fork length. Increasing dietary lipid level or including vegetable oil, RO, in the feeds did not increase liver lipid content. Liver fatty acid compositions largely reflected dietary profiles confirming very limited endogenous LC-PUFA biosynthesis. Liver of ABT fed 15KO and 20KO displayed the highest contents of docosahexaenoic acid (DHA). The hepatic expression of genes encoding enzymes and transcription factors involved in lipid and fatty acid metabolism, as well as genes encoding antioxidant enzymes, showed that many of these genes were regulated by dietary lipid and LC-PUFA content. Results suggested that ABT juveniles can be on-grown on inert dry feeds that support good fish growth and the accumulation of DHA.

Effects of age on growth in Atlantic bluefin tuna (Thunnus thynnus).

Atlantic Bluefin Tuna Thunnus thynnus (ABFT) is considered one of the most important socio-economic species but there is a lack of information on the physiological and molecular processes regulating its growth and metabolism. In the present study, we focused on key molecules involved in growth process. The aim of the present study was to associate molecular markers related to growth with canonical procedures like morphological measurements such as curved fork length (CFL) and round weight (RWT). The ABFT specimens (n = 41) were organized into three different groups A, B and C according to their age. The molecular analysis of liver samples revealed that igf1, igf1r and mTOR genes, involved in growth process, were differentially expressed in relation to the age of the fish. In addition, during the analyzed period, faster growth was evident from 5 to 8 years of age, after that, the growth rate decreased in terms of length yet increased in terms of adipose tissue storage, as supported by the higher fat content in the liver. These results are useful in expanding basic knowledge about the metabolic system of ABFT and provide new knowledge for the aquaculture industry.

Expression analysis of the Atlantic bluefin tuna (Thunnus thynnus) pro-inflammatory cytokines, IL-1ß, TNFα1 and TNFα2 in response to parasites Pseudocycnus appendiculatus (Copepoda) and Didymosulcus katsuwonicola (Digenea).

Pro-inflammatory cytokines play an important role in teleost defence against numerous types of pathogens, therefore are often used as biomarkers during various infections. In order to evaluate Atlantic bluefin tuna IL-1ß, TNFα1 and TNFα2 induction by PAMPs, we quantified their expression during in vitro stimulation of peripheral blood leukocytes by LPS and Poly I:C. Furthermore, their role in acute and chronic parasitic infection was examined during natural infection of Pseudocycnus appendiculatus (Copepoda) and Didymosulcus katsuwonicola (Digenea), as well as during leukocyte exposure to total protein extracts isolated from two parasite species. Induction of ABT IL-1ß and TNFα2 by PAMPs and protein extracts from D. katsuwonicola and P. appendiculatus, as well as during natural infection with two parasites, suggests these cytokines play an important role in inflammation, being engaged in controlling parasite infections, in contrast to ABT TNFα1. Cellular innate response to the digenean D. katsuwonicola showed rather chronic character, resulting with parasite encapsulation in connective tissue. Mast cells, eosinophils, goblet cells, and occasional rodlet cells found at the site of infection, along with the induction of TNFα2, suggest the presence of a moderate inflammatory reaction that fails to seriously endanger digenean existence. In contrast, copepod P. appendiculatus, attached to the gill epithelium by clamping, caused direct tissue disruption with undergoing necrotic or apoptotic processes, and extensive proliferation of rodlet and goblet cells. Differential expression patterns of target cytokines in tissue surrounding two parasites and in vitro PBL model suggest that quality and quantity of tuna immune response is conditioned by parasite adaptive mechanisms and pathogenicity.

Lead and cadmium in blood and tissues of Atlantic bluefin tuna (Thunnus thynnus L., 1758).

Cadmium (Cd) and lead (Pb) levels in blood and tissues of Atlantic bluefin tuna were analysed to gather information regarding their distribution, accumulation and inter-relationships, as well as to examine how sex affects them. In the whole population, the concentration range was from below the detection limit (bone) to 8.512 µg g-1 (liver) for Cd, and from below detection limit (bone and gills) to 0.063 µg g-1 (kidney) for Pb. The median concentration in the muscles (0.008 and 0.029 µg g-1 for Cd and Pb, respectively) was 10 times less than the maximum permitted for consumption. Sex was shown to be an important variable affecting concentrations of Cd in both liver and kidneys, so taking into account sex when interpreting results is highly recommended. The importance of Cd and Pb bioaccumulation in fishery by-products, increasingly important in commercial circuits, is also highlighted.

Otolith-temperature estimates in Atlantic bluefin tuna (Thunnus thynnus) from the Mediterranean Sea: Insights from clumped isotope measurements.

The subpopulation and/or contingent structure of Atlantic bluefin tuna (Thunnus thynnus) within the Mediterranean Sea is undefined, leading to uncertainty regarding the best strategy for an effective assessment and management of this highly exploited stock. This study aimed to reconstruct temperatures experienced by Atlantic bluefin tuna during the early life period (<3.5 months) using clumped isotope temperature proxy, an innovative geothermometer for carbonates, that does not require previous knowledge of other environmental parameters such as water oxygen composition. We examined otolith chemistry in fish captured from 3 different areas of the Mediterranean Sea and adjacent waters. We found that mean seasonal temperature estimates from clumped isotopes did not differ significantly from satellite derived and otolith oxygen stable isotopic ratios derived temperatures, except for the central Mediterranean Sea, were clumped isotopes derived temperatures were significantly higher than satellite derived temperatures. However, the sensitivity of the clumped isotope thermometer was found to be lower than that based on oxygen fractionation equation, with high variance observed in the clumped isotopes derived temperature estimates. We also observed that clumped isotope derived temperatures were undistinguishable among bluefin tuna captured in the Gibraltar Strait, the central, and eastern Mediterranean Sea. In this paper, we discuss the major sources of uncertainty in temperature reconstructions using bluefin tuna otoliths.

Maternal Effects and Trophodynamics Drive Interannual Larval Growth Variability of Atlantic Bluefin Tuna (Thunnus thynnus) from the Gulf of Mexico.

Two cohorts of Atlantic bluefin tuna (Thunnus thynnus) larvae were sampled in 2017 and 2018 during the peak of spawning in the Gulf of Mexico (GOM). We examined environmental variables, daily growth, otolith biometry and stable isotopes and found that the GOM18 cohort grew at faster rates, with larger and wider otoliths. Inter and intra-population analyses (deficient vs. optimal growth groups) were carried out for pre- and post-flexion developmental stages to determine maternal and trophodynamic influences on larval growth variability based on larval isotopic signatures, trophic niche sizes and their overlaps. For the pre-flexion stages in both years, the optimal growth groups had significantly lower δ15N, implying a direct relationship between growth potential and maternal inheritance. Optimal growth groups and stages for both years showed lower C:N ratios, reflecting a greater energy investment in growth. The results of this study illustrate the interannual transgenerational trophic plasticity of a spawning stock and its linkages to growth potential of their offsprings in the GOM.

Intra-pituitary relationship of follicle stimulating hormone and luteinizing hormone during pubertal development in Atlantic bluefin tuna (Thunnus thynnus).

As part of the endeavor aiming at the domestication of Atlantic bluefin tuna (BFT; Thunnus thynnus), first sexual maturity in captivity was studied by documenting its occurrence and by characterizing the key hormones of the reproductive axis: follicle stimulating hormone (FSH) and luteinizing hormone (LH). The full length sequence encoding for the related hormone ß-subunits, bftFSHß and bftLHß, were determined, revealing two bftFSHß mRNA variants, differing in their 5' untranslated region. A quantitative immuno-dot-blot assay to measure pituitary FSH content in BFT was developed and validated enabling, for the first time in this species, data sets for both LH and FSH to be compared. The expression and accumulation patterns of LH in the pituitary showed a steady increase of this hormone, concomitant with fish age, reaching higher levels in adult females compared to males of the same age class. Conversely, the pituitary FSH levels were elevated only in 2Y and adult fish. The pituitary FSH to LH ratio was consistently higher (>1) in immature than in maturing or pubertal fish, resembling the situation in mammals. Nevertheless, the results suggest that a rise in the LH storage level above a minimum threshold may be an indicator of the onset of puberty in BFT females. The higher pituitary LH levels in adult females over males may further support this notion. In contrast three year-old (3Y) males were pubertal while cognate females were still immature. However, it is not yet clear whether the advanced puberty in the 3Y males was a general feature typifying wild BFT populations or was induced by the culture conditions. Future studies testing the effects of captivity and hormonal treatments on precocious maturity may allow for improved handling of this species in a controlled environment which would lead to more cost-efficient farming.

Atlantic bluefin tuna diet variability in the southern Gulf of St. Lawrence, Canada.

The abundance of top predators in the southern Gulf of St. Lawrence, Canada, has fluctuated dramatically in recent decades. The associated increase in predation and its effect on the lack of recovery of many fish stocks in the system generates the need for a better understanding of predator-prey relationships and the implementation of an ecosystem approach to fisheries management. This study used stomach content analysis to further describe the diet of Atlantic bluefin tuna in the southern Gulf of St. Lawrence. Teleost fish largely dominated the stomach contents in all years. Previous studies established that Atlantic herring was the main component of the diet by weight, whereas herring was almost absent from the diet in this study. A shift in the diet of Atlantic bluefin tuna has been observed, as it now feeds almost exclusively on Atlantic mackerel. The estimated daily meal varied between years, ranging from 1026 g per day in 2019 to 2360 g per day in 2018. Daily meals and daily rations were calculated and showed substantial year-to-year variation.

Effects of Age, Fulton's Condition Index (K) and Muscle Fat on Total Mercury Content in Raw, Pre-Canning and Canned Samples of Atlantic Bluefin Tuna (Thunnus thynnus).

A total of 30 samples of Atlantic bluefin tuna were analysed for total mercury concentration. Relationships between total mercury content and age, Fulton's condition index (K) and fat content were statistically evaluated. The effect of muscle status (raw, pre-canning, canned) on mercury content was also investigated. The average total mercury content was: 1.185 ± 0.662 mg/kg in raw, 1.481 ± 0.893 mg/kg in pre-canning and 1.393 ± 0.882 mg/kg in canned samples, respectively. Canning promotes a statistically significant increase in the concentration of mercury. The weight of fish, K and fat content are useful tools to estimate the mercury accumulation in Atlantic bluefin tuna. The results of the present study represent a contribution to the assessment of the EU mercury levels in Atlantic bluefin tuna.

Evidence of isotopic maternal transmission influence on bluefin tuna (Thunnus thynnus) larval growth.

Pre-flexion stages of Atlantic bluefin tuna (Thunnus thynnus) larvae were collected in 2014 during the peak of spawning in the two main spawning areas: Gulf of Mexico (GOM) and Mediterranean Sea (MED). We examined daily growth, otolith biometry, and stable isotopes and found that the GOM grew at a faster rate, had larger otoliths, wider daily increments, and significantly lower values of δ15N when compared to the MED. In addition, an intra-population comparative analysis between slow- and fast-growing individuals (deficient vs. optimal growth groups, respectively) showed that optimal growth groups had significantly lower δ15N within each spawning area, implying a direct relationship between growth potential, development, and maternal transmission of isotopic signatures. A third pre-flexion larval group that was aquaculture-reared also exhibited the same pattern to the wild larval groups. In addition, for the first time, we estimated the maternal trophic niches using models developed with field-captured pre-flexion larvae. The estimated maternal trophic niches for the GOM were narrower than the MED, implying differences in the maternal trophodynamics from each nursery area. Overall, the inter-population (GOM vs. MED) and intra-population growth groups (deficient vs. optimal) grew faster and had narrower maternal niches. This study shows the advantages that larval SIA research can aid in the understanding of the trophodynamics of their breeders by examining the trophic relationship of a spawning stock jointly with the development of growth potential in offspring within the same breeding season.

Histogenesis and cell differentiation in the retina of Thunnus thynnus: A morphological and immunohistochemical study.

This study examines the anatomical development of the visual system of Atlantic bluefin tuna, Thunnus thynnus, during the first 15 days of life at histological level, with emphasis in the immunohistochemical characterization of different cell types. As an altricial fish species, the retina was not developed at hatching. The appearance of eye pigmentation and the transformation of the retina from an undifferentiated neuroblastic layer into a laminated structure occurred during the first two days of life. At 16 days after hatching (DAH), the ganglion cells were arranged in a single row in the central region of the retina and the outer segments of the photoreceptors were morphologically developed. Furthermore, at this age, all the retinal cell types were immunohistochemically characterized. The presence of ganglion cell axons was confirmed with the TUJ1 antibody and the existence of functional synapses in the plexiform layers with antibodies against SV2. Cone opsins were immunostained with antibodies against visinin and CERN-922 immunoreactive rods were also identified. Different subpopulations of amacrine cells were immunostained with antibodies against αTH and PV. Highly GS-immunoreactive Müller cells were also detected at this age. These observations suggested that the T. thynnus retina was fully functional at the end of the second week of life. Basic studies on early morphology of the visual system and larval behavior are necessary to support applied research on larval rearing. Furthermore, they may have implications for understanding larval ecology in the wild.

Chemical signatures in fin spine edge of Atlantic bluefin tuna (Thunnus thynnus) can serve as habitat markers of geographically distinct marine environments.

Chemical fingerprints in otoliths are commonly used as natural habitat markers in fishes. Alternatively, the first dorsal fin spine can provide valuable chemical information and may be more suitable for studying (i) endangered fish species that cannot be sacrificed for their otoliths or (ii) fishes for which otoliths might not be available because of management or commercial reasons. Here, we studied multi-element chemistry of fin spine edges collected from Atlantic bluefin tuna (ABFT; Thunnus thynnus) (Linnaeus, 1758) to investigate the utility of the fin spine edge as a natural habitat marker. We determined stable isotopic δ18O and δ13C ratios, as well as concentrations of the tracer elements Mg, Mn, Li, Ba, and Sr, at the edge of ABFT fin spines, and then we used these measures to discriminate ABFT individuals among capture regions (i.e., the eastern Atlantic Ocean or Mediterranean Sea). Isotope ratios and tracer element concentrations, and especially a combined multi-element approach, were able to effectively discriminate individuals by capture region. The Mg, Mn, Li, and δ18O concentrations were the strongest variables driving this discrimination. Overall, our results demonstrate that chemical signatures are consistently retained in the ABFT fin spine edge and support the use of fin spine edges for discerning habitat use. The fin spine chemistry as a minimally invasive sampling method, combined with otolith chemistry, genetic markers, and tagging efforts can help us to reconstruct fish movements, providing a deeper understanding of the spatial population dynamics of this iconic fish species.