Development of melanin-concentrating hormone-immunoreactive elements in the brain of gilthead seabream (Sparus auratus)

The development of the hypothalamic melanin-concentrating hormone (MCH) system of the teleost Sparus auratus has been studied by immunocytochemistry using an anti-salmon MCH serum. Immunoreactive perikarya and fibers are found in embryos, larvae, and juvenile specimens. In juveniles, most labeled neurons are present in the nucleus lateralis tuberis; some are dispersed in the nucleus recessus lateralis and nucleus periventricularis posterior. From the nucleus lateralis tuberis, MCH neurons project a conspicuous tract of fibers to the ventral hypothalamus; this penetrates the pituitary stalk and reaches the neurohypophysis. Most fibers end close to the cells of the pars intermedia, and some reach the adenohypophysial rostral pars distalis. Immunoreactive fibers can also be seen in extrahypophysial localizations, such as the preoptic region and the nucleus sacci vasculosi. In embryos, MCH-immunoreactive neurons first appear at 36 h post-fertilization in the ventrolateral margin of the developing hypothalamus. In larvae, at 4 days post-hatching, perikarya can be observed in the ventrolateral border of the hypothalamus and in the mid-hypothalamus, near the ventricle. At 26 days post-hatching, MCH perikarya are restricted to the nucleus lateralis tuberis. The neurohypophysis possesses MCH-immunoreactive fibers from the second day post-hatching. The results indicate that MCH plays a role in larval development with respect to skin melanophores and cells that secrete melanocyte-stimulating hormone. Received: 4 April 1995 / Accepted: 17 July 1995     

Reproduction and early development of a freshwater pipefish <Emphasis Type="Italic">Microphis leiaspis</Emphasis> in Okinawa-jima Island,Japan

We investigated the size at maturation, breeding season, and morphological development of larvae and juveniles of a freshwater pipefish Microphis leiaspis, which belongs to Gastrophori, collected from three rivers on the northern part of Okinawa-jima Island, Japan. The minimum size of brooding males was 105–123 mm in standard length (SL). The smallest mature female was estimated to be ca. 130 mm SL from the analysis of gonadosomatic index (GSI) and histological observations of gonads. The breeding season was estimated to be from June to December according to monthly changes in female GSI, histological observations of gonads, and monthly changes in the occurrence of brooding males. The number of eggs in the male brood pouch ranged from 75 to 241 (mean ± SD: 152 ± 52, n = 22). The male releases newly hatched larvae in freshwater areas. After newborns grow in the sea, they return to freshwater areas of the rivers and attain maturity. Microphis leiaspis was conformed to have an amphidromous life history. Notochord length of the released larvae was 6.1 mm, with a well-developed finfold. Larvae attained 11.1 mm SL, formation of the caudal and dorsal fin rays was complete, and the caudal fin became lozenge shaped at 30 days after the release, and juveniles reached 36.0 mm SL at 63 days after release. In the period between 30 and 63 days after the release, formation of all fins except the pectoral fins was completed, and caudal fin rays were extended and sector shaped with deep slits between each fin ray. The morphology of the released larvae of M. leiaspis is similar to that of Gastrophori species, and the morphology of juveniles similar to other species of Microphis.     

Carp (Cyprinus carpio L.) innate immune factors are present before hatching

Expression of the innate immune factors, complement factor 3 (C3), alpha2-macroglobulin (alpha2M), serum amyloid A (SAA) and a complement factor 1 r/s--mannose binding lectin associated serine protease-like molecule (C1/MASP2), was determined with Real Time Quantitative-PCR in carp (Cyprinus carpio L.) ontogeny around hatching. Furthermore, the expression of C3 mRNA and the presence of C3 protein were studied in carp embryos and larvae using In Situ Hybridisation, Western Blotting and Immunohistochemistry. C3, alpha2M, SAA and C1/MASP2 mRNA were produced by embryos from 12 h post-fertilisation, which is relatively long before hatching (2 days post-fertilisation (dpf)), indicating either involvement of these factors in development itself or more probably a preparation of the immune system for the post-hatching period. In addition, maternal mRNA of the aforementioned innate immune factors and maternal C3- and immunoglobulin protein was present in unfertilised eggs. Furthermore, C3 mRNA production was situated in the yolk syncytial layer in embryos from 24 h post-fertilisation to 5 dpf, followed by the liver in larvae, providing a new sequence of C3 production in teleost development.     

Determining the optimum temperature and salinity for larval culture,and describing a culture protocol for the conservation aquaculture for European smelt Osmerus eperlanus (L.)

Populations of anadromous European smelt Osmerus eperlanus (L.) are declining across its geographical range in northern Europe, but no practical culture techniques exist to develop stock enhancement programmes for this species. In this study, a culture protocol is described to rear fish from fertilised eggs to mature adults in 2 years involving the use of ‘green water’, live feed and artificial diets. The sequence of embryonic development for eggs incubated at 10°C/0 ppt was described and photographed. To determine the optimum conditions for larval culture, fertilised eggs were reared at a range of salinities (0–20 ppt) and temperatures (5–18°C) until first feeding. Best hatching success (ca. 97%), size at hatch (ca. 0.8 mm) and survival to first feeding (ca. 96%) of larvae were achieved under combined conditions of low salinity (0–0 ppt) and temperature (5–10°C). No larvae survived a salinity of 20 ppt. The time taken from fertilisation to hatch (FtH) and hatching duration (HD) were temperature-dependent ranging from 42 days FtH and 10 days HD at 5ºC, to 10 days FtH and 2 days HD at 18°C irrespective of salinity. The results indicate that conservation programmes could utilise existing salmonid hatchery facilities (i.e. freshwater, ≤10°C water temperature) for stock enhancement. Since on-growing of smelt involves the logistical and technical problems of live feed production, it is recommended that smelt enhancement programme utilise freshwater hatchery facilities to rear fish until hatching, and then stock out onto known spawning grounds in rivers allowing hatched larvae to drift into estuaries to complete the larval and juvenile phases. This approach would minimise the time spent in the hatchery post-hatching, eliminate the need for live food production, prevent the development of predator-naïve fish, and hence would mimic the natural life cycle of the species as closely as possible.     

The development of the stomach in Clarias lazera and the intestinal absorption of protein macromolecules

Summary The development of the stomach of the teleost, Clarias lazera, during the early posthatching period, is described, and the developing stomach is compared with that of adult Clarias.The stomach develops in two distinct parts: the corpus, which differentiates first, and the pylorus. The corpus contains a mucous surface epithelium, arranged in folds, and a tubular gland system containing only one type of gland cell, to which the secretion of pepsinogen and HCl is attributed. The pyloric region does not contain tubular glands.From the ultrastructure of the gland cells, the ³H-thymidine labeling index, and the onset of acid production (as determined with pH indicators) it is concluded that a functional stomach is present in juveniles with a standard length of ± 11 mm (approximately 12 days after fertilization at 23–24° C).The ultrastructure of the intestinal epithelium has also been studied. The intestine consists of three segments, similar to those described for stomachless teleosts and a number of fish larvae. In larvae as well as in juveniles, the enterocytes of the second segment show pinocytosis of horseradish peroxidase, although in the juveniles the stomach has already developed. This second segment has the same relative length in all studied larvae and juveniles and is also present in adult Clarias.It is therefore concluded that the capacity to absorb protein macromolecules is not specifically related to the absence of a functional stomach in this teleost species.     

Early development of the endemic dace Telestes karsticus (Leuciscidae,Cypriniformes) in a Dinaric karst stream in Croatia

Early development of the endemic dace Telestes karsticus is described in a Dinaric karst stream in Croatia. Larvae were acquired from successful rearing in an aquarium. Basic morphometric and meristic measures of larvae were taken during 55 days of observation. The mean total length of larvae at the time of hatching was 6.45 mm. The yolk sac was completely absorbed in all larvae by day 9 post-hatching (PH), when the mouth also opened and independent feeding and intestinal function were observed. Flexion of the urostyle was completed between days 12–16 PH. The caudal fin became completely forked on day 23 PH. Formation of fin rays was complete on day 48 PH when fish fully resembled juvenile specimens. The number of soft ray fins corresponds to the adult. Understanding the early development is particularly valuable in assessing environmental impacts and recovering of this endangered species and it is essential for effective monitoring and its conservation. This is the first report of the early development for the endemic Telestes karsticus, pointing out the importance of the obtained results for the effective conservation of the species.     

Changes in marine turbot (Scophthalmus maximus) epidermis and skin mucus composition during development from bilateral larvae to juvenile flat fish

Alike other flat fish, marine turbot has the particularity that changes from larvae with bilateral symmetry to adult with asymmetry, in terms of the position of the eyes. As expected, the skin configuration of this species is also affected by the development and transformation suffered by fish during metamorphosis. In this context, changes in the epidermis of marine turbot were studied using conventional staining and histochemical techniques using six lectins (UEA-I, PNA, RCA-I, WGA, Con A and SBA). During development from larvae to juvenile (3–300 days post-hatching), the epidermis increased in both thickness and the number of cell layers. In fact, the simple cuboidal epithelium observed in larvae at day 3 already became stratified at days 10–12, which sequentially increase in thickness with fish development. Turbot epidermis is composed basically of four cell types: epithelial and mucous or secretory cells that are present through the development, and pigmented cells and a type that the authors described as club-like cells that appear during and post-metamorphosis. The Alcian blue-periodic acid Schiff (AB-PAS) histochemical method revealed the presence of neutral glycoconjugates in mucous and club-like cells at post-metamorphic stages of fish. Accordingly, lectin analysis showed mucous cells containing glycoproteins rich in fucose (UEA-I labelling) and glycoconjugates rich in the sequence galactose-N-acetyl galactosamine (PNA and RCA-I labelling) when this cell type appears. Interestingly, melanophores were observed in the dorsal epidermis of post-metamorphic juveniles. This type of cell contains a black-to-brown pigment that provides the skin the typical colour of this fish species. Changes in mucous coat composition were observed during fish development, which was attributed to different roles of the glycoconjugates.     

Molecular cloning and expression analysis of a F-type lectin gene from Japanese sea perch (<Emphasis Type="Italic">Lateolabrax japonicus</Emphasis>)

The techniques of homology cloning and anchored PCR were used to clone the fucose-binding lectin (F-type lectin) gene from Japanese sea perch (Lateolabrax Japonicus). The full-length cDNA of sea perch F-lectin (JspFL) contained a 5′ untranslated region (UTR) of 39 bp, an ORF of 933 bp encoding a polypeptide of 310 amino acids with an estimated molecular mass of 10.82 kDa and a 3′ UTR of 332 bp. The searches for nucleotides and protein sequence similarities with BLAST analysis indicated that the deduced amino acid sequence of JspFL was homological to the Fucose-binding lectin in other fish species. In the JspFL deduced amino acid sequence, two tandem domains that exhibit the eel carbohydrate-recognition sequence motif were found. The temporal expressions of gene in the different tissues were measured by real-time PCR. And the mRNA expressions of the gene were constitutively expressed in tissues including spleen, head-kidney, liver, gill, and heart. The JspFL expression in spleen was different during the stimulated time point, 2 h later the expression level became up-regulated, and 6 h later the expression level became down-regulated. The result indicated that JspFL was constitutive and inducible expressed and could play a critical role in the host-pathogen interaction.     

长江口水域夏季鱼卵和仔稚鱼年间变化

基于2005年、2008年、2009年和2011年8月(夏季)在长江口水域(30°30'—31°45'N,121°15'—123°10'E)4个航次的浮游生物拖网资料,分析了长江口水域鱼卵和仔稚鱼的种类组成、数量分布特征及其年间变化。结果表明:4个航次采集的鱼卵和仔稚鱼鉴定到种的种类有17种,隶属于8目13科,以鲈形目种类最多,11种,其次是鲱形目,5种,其他各目种类均小于5种;种类数存在明显年间差异,2005年种类数最多(鱼卵3种,仔稚鱼8种),其次是2009年和2011年,2008年种类数最少(鱼卵1种,仔稚鱼5种)。优势种年间更替明显,长蛇鲻(Saurida elongata)、虾虎鱼(Gobiidae spp.)和中华小公鱼(Stolephorus chinensis)在2005年是优势种,2008年优势种为鳀鱼(Engraulis japonicus),2009年优势种为鳀鱼、寡鳞飘鱼(Pseudolaubuca engraulis)、虾虎鱼等,2011年虾虎鱼和小公鱼(包括小公鱼属未定种Stolephorus spp.和中华小公鱼Stolephorus chinensis)成为优势种。2005年鱼卵和仔稚鱼数量分布的密集区在嵊泗列岛附近水域,2008年鱼卵和仔稚鱼出现较少,未出现明显的数量密集区;2009年鱼卵数量较少,仔稚鱼数量较多,密集区主要分布在在长江口以外123°E附近水域;2011年鱼卵主要分布在在长江北支口门外附近水域,仔稚鱼在调查区内分布相对均匀。     

Ontogenetic behavior of Kootenai River White Sturgeon, <Emphasis Type="Italic">Acipenser transmontanus</Emphasis>, with a note on body color: A laboratory study

Laboratory studies indicated the following ontogenetic behavior and body color of wild Kootenai River White Sturgeon, Acipenser transmontanus, (hereafter, Kootenai Sturgeon), a landlocked population in the Kootenai River, a major tributary of the Columbia River (United States) and Kootenay Lake (Canada). Hatchling free embryos (hereafter, embryos) are photonegative and hide under cover at a spawning site, and have a grey body. Late-embryos are photopositive and weakly prefer white substrate, use cover less with age, and develop a black tail. Day 13 larvae forage in the day on the open bottom, use cover less with age, prefer bright habitat, have a light-grey body and black tail, and initiate a mostly nocturnal dispersal for about 21 days, and then, continue a weaker dispersal. As they age, the entire body and tail of larvae is a dark-grey color when they develop into juveniles (about 66 days). The common body and tail color of larvae from the Kootenai, Columbia, and Sacramento rivers indicate a common adaptation to signal conspecifics or avoid predators. Juveniles are variable for foraging height, do not hide in bottom cover, and continue a weak nocturnal downstream movement. Movement of larvae and juveniles in the artificial stream suggests wild Kootenai Sturgeon have a long slow dispersal style (disperse for months). The long dispersal style of young Kootenai Sturgeon may adapt larvae to dispersing all summer in a 100–200 km long reach with a low abundance of food. The final destination of Kootenai Sturgeon during their first rearing season is unknown, but the long dispersal suggests fish could easily move to the lower river or to Kootenay Lake. Ontogenetic behavior of Kootenai Sturgeon is slightly different from Columbia River White Sturgeon, which has a weak embryo dispersal, but both populations have a similar major dispersal by larvae. However, both of these populations differ qualitatively from Sacramento River White Sturgeon, in which juveniles initiate the major dispersal. Thus, major geographic behavioral variation exists among populations and should be considered in restoration programs.     

Comparative survival and growth of embryos,larvae, and juveniles of pejerrey Odontesthes bonariensis and O. hatcheri* at different salinities

The hatching of fertilized eggs and the survival and growth of larvae and juveniles of the inland‐water atherinids Odontesthes bonariensis (Valenciennes 1835) and O. hatcheri (Eigenmann 1909) were examined at salinities of 0, 5, 10, 20, and 30 ppt. In addition, a limited study compared the salinity responses of O. bonariensis eggs and larvae from different origins. Overall, embryos, larvae, and juveniles of both species were euryhaline, although best survival and growth rates were obtained at the intermediate salinities. Survival of O. bonariensis at 0 ppt varied from very good to very poor. Comparison of the salinity responses of eggs and larvae of O. bonariensis from the current Japanese strain with newly introduced strains from three locations in Argentina did not reveal a clearly superior strain for freshwater culture. In general, O. hatcheri showed higher survival and growth rates and better adaptability to fresh water compared with O. bonariensis. Although both species are commonly regarded as freshwater species, the results of this study emphasize the importance of millimolar quantities of salts in the rearing water for improved survival and growth.     

The early life history of kurosoi,Sebastes schlegeli (Scorpaenidae), in the Sea of Japan

Larval and juvenile stages of kurosoi,Sebastes schlegeli, are described and illustrated from wild specimens. Some ecological aspects of larvae and juveniles are also described. Notochord flexion occurred between 5.6–7.5 mm SL. Transformation occurred between 13–20 mm SL. Preflexion and flexion larvae ofS. schlegeli can be distinguished from similar larvae by the pigmentation of the dorsal and ventral midlines of the tail and absence of pigmentation on the ventral portion of the rectum. After notochord flexion, the dorsal and lateral regions in both larvae and pelagic juveniles were heavily pigmented, suggesting adaptation for neustonic life style. Larvae and juveniles were caught at many coastal stations, but did not occur in cooler offshore waters. Larvae smaller than 20 mm SL inhabited surface waters. Until ca. 40 mm SL, juveniles inhabited mainly surface waters (without drifting seaweed), but also used other habitats, such as the drifting seaweed, and near the sea bed. Small larvae (<7 mm SL) fed mainly on copepod nauplii. Larger larvae fed on calanoid copepodites andEvadne nordmanni. Pelagic juveniles fed mainly on fish eggs, with fish larvae also being important food items for some individuals. Most food items taken by juveniles that were associated with drifting seaweed were eggs with attaching filaments (Cololabis saira andHyporhamphus sajori), suggesting that the high density of such food items both attracts and keeps juveniles around drifting seaweed.     

High stocking density during larviculture and effect of size and diet on production of juvenile Lophiosilurus alexandri Steindachner, 1876 (Siluriformes: Pseudopimelodidae)

Different stocking densities were investigated in larviculture and feeding of Lophiosilurus alexandri, as well as analyses of the effects on juveniles of two size‐classes and two different commercial formulated diets. The first experiment was two‐phased: (a) larvae stocked at densities of 60, 120, 180, 240, and 300 larvae L⁻¹ fed with Artemia nauplii and reared for 15 days; (b) in phase 2, densities of 5, 10, 15, 30, and 40 juveniles L⁻¹ were evaluated during feed training (20 days). Mean water temperature in both phases was 28°C. In the first phase of experiment 1, the different stocking densities did not affect fish growth or survival. In phase 2, growth was similar in all densities; however, survival was lower at higher densities. The increased density provided a rise in biomass and number of individuals produced in both phases. In the second experiment, two size‐classes of feed‐trained juveniles (30.22 ± 1.84 and 34.66 ± 2.41 mm) were given pellets of two different diameters (1.2 and 2.6 mm) for 20 days. The largest juveniles fed the 1.2 mm inert diet had higher final weights and lengths. Larviculture and feed training of L. alexandri can thus be performed successfully at high stocking densities of 300 larvae L⁻¹ during the first 15 days of feeding, and at densities of up to 40 juveniles L⁻¹ during the 20 days of feed training, respectively.     

Embryonic development and larval behavior of the kissing loach (<Emphasis Type="Italic">Parabotia curta</Emphasis>): adaptations to an ephemeral,hypoxic environment

Many teleost fishes in lowland fresh waters spawn in ephemeral flooded areas, the bottoms of which are prone to hypoxia. Little is known about how embryos and larvae deal with these potentially hostile environments. This study examines the functional and behavioral ontogeny of one such species, the kissing loach (Parabotia curta). Kissing loach eggs are demersal and adhesive. Hatching occurs at 24.8 ± 0.1 h post-fertilization at 25°C, much earlier than most fish species. The newly hatched larvae are precocious with no functional mouth, fins or eye pigmentation. Swimbladder inflation normally occurs at about 4 days posthatch, even before which the hatched larvae moved immediately toward the water surface to hang from water moss. Experiments with larvae 20 h after hatching showed that they spent significantly less time on the bottom in hypoxic water (2 mg/l) than in normoxic water, and suggest that hypoxia is a major directive factor in eliciting surfacing behavior. For the kissing loach, we have previously reported short-term spawning after the formation of flood areas as well as wide scattering of the spawned eggs in the temporal flooded areas. These traits with the present results of hatching at an early stage and the immediate upward movement of larvae are considered to be effective strategies for using ephemeral, hypoxic flooded areas for reproduction.     

Behavior of early life intervals of Klamath River green sturgeon, <Emphasis Type="Italic">Acipenser</Emphasis><Emphasis Type="Italic">medirostris</Emphasis>, with a note on body color

We studied ontogenetic behavior, migration, and wintering behavior of young Klamath River green sturgeon, Acipenser medirostris, in the laboratory to provide insight into likely behavior of wild sturgeon. Hatchling free embryos preferred cover but were poor swimmers and could not move farther than a few centimeters to cover. The poor swimming ability and cover preference of hatchlings suggests evolution for habitat selection of females to place eggs in habitat with cover for eggs (and hatchlings), and for egg characteristics (large, dense, and weakly adhesive) to cause rapid sinking into cover without drifting. A day or so after fish developed into larvae (first life interval feeding exogenously), day-12 larvae initiated a 12-day downstream nocturnal migration. A totally nocturnal migration is unlike other Acipenser migrants yet studied. Migrant larvae had a dark-colored body typical of other Acipenser species that migrate as larvae. Tail color was a dark black (black-tail phenotype) only during the early larva period, suggesting a morphological adaptation for migration, foraging, or both. Post-migrant larvae and early juveniles to day 84 foraged diurnally with a nocturnal activity peak. Day 110–181 juveniles moved downstream at night until water temperature decreased to about 8°C, indicating wild juveniles migrate downstream to wintering habitat. Habitat preference of month 9–10 wintering juveniles suggests wild juveniles are in deep pools with low light and some rock structure. Wintering juveniles were only active at night. Initiation and cessation of daily activity was at dusk and dawn during illumination changes of <1.0lx. This sensitivity to illumination has not been found before in sturgeons. During the first 10months of life, nocturnal activity of early life intervals is a dominant feature of migration, foraging, and wintering.     

Investigating the developmental onset of regenerative potential in the annelid Capitella teleta

An animal's ability to regrow lost tissues or structures can vary greatly during its life cycle. The annelid Capitella teleta exhibits posterior, but not anterior, regeneration as juveniles and adults. In contrast, embryos display only limited replacement of specific tissues. To investigate when during development individuals of C. teleta become capable of regeneration, we assessed the extent to which larvae can regenerate. We hypothesized that larvae exhibit intermediate regeneration potential and demonstrate some features of juvenile regeneration, but do not successfully replace all lost structures. Both anterior and posterior regeneration potential of larvae were evaluated following amputation. We used several methods to analyze wound sites: EdU incorporation to assess cell proliferation; in situ hybridization to assess stem cell and differentiation marker expression; immunohistochemistry and phalloidin staining to determine presence of neurites and muscle fibers, respectively; and observation to assess re-epithelialization and determine regrowth of structures. Wound healing occurred within 6 h of amputation for both anterior and posterior amputations. Cell proliferation at both wound sites was observed for up to 7 days following amputation. In addition, the stem cell marker vasa was expressed at anterior and posterior wound sites. However, growth of new tissue was observed only in posterior amputations. Neurites from the ventral nerve cord were also observed at posterior wound sites. De novo ash expression in the ectoderm of anterior wound sites indicated neuronal cell specification, although the absence of elav expression indicated an inability to progress to neuronal differentiation. In rare instances, cilia and eyes re-formed. Both amputations induced expanded expression of the myogenesis gene MyoD in preexisting tissues. Our results indicate that amputated larvae complete early, but not late, stages of regeneration, which indicates a gradual acquisition of regenerative ability in C. teleta. Furthermore, amputated larvae can metamorphose into burrowing juveniles, including those missing brain and anterior sensory structures. To our knowledge, this is the first study to assess regenerative potential of annelid larvae.     

Occurrence and morphology of larvae and juveniles of six <Emphasis Type="Italic">Luciogobius</Emphasis> species from Aritsu Beach,Okinawa Island

Larvae and juveniles of six species of Luciogobius were collected at Aritsu Beach on Okinawa Island using a small seine. Postflexion larvae were dominant during sampling and were collected when they approached the shoreline adjacent to or at the entrances to their adult habitats prior to settlement. Standard lengths of postflexion larvae ranged from 5.4 to 14.4 mm and varied depending on the species. The larvae occurred mainly from January to April, but some larvae were caught in October and November. Their pelagic larval durations were estimated to range from 17 to 36 days and varied depending on the species. Morphologies of field-caught larvae and juveniles and laboratory-reared juveniles were described. Six species were clearly distinguished based on fin ray and vertebral counts, proportions, body size, and pigment patterns. Although their taxonomic statuses could not be determined, it is thought that they have independent relatives in other regions.     

Ontogeny of enzymatic activities in fed and fasting turbot, Scophthalmus maximus L.

The presence and localization of acid and alkaline phosphatase, non-specific proteases, aminopeptidase, amylase, non-specific esterase and lipase was investigated by histoenzymologic methods in fed and fasting turbot from day 1 to day 40 post-hatching and compared with published data. Alkaline phosphatase and aminopeptidase activities were delected at day 1 in the distal region of the developing digestive tube. At day 3 (opening of the mouth) aminopeptidase and alkaline phosphatase activities were found all along the intestine. Sites of non-specific esterase and protease activities became apparent in the digestive tract at days 2 and 3 respectively. Amylase was present in the exocrine pancreas at day 3 and in the lumen of the intestine at day 4. Acid phosphatase was active in the cellular structure surrounding the yolk stores and in the lipid droplets at day 1 and in the intestinal epithelium at day 3. Lipase was found at day 15 when the larvae metamorphose into juveniles.
All the investigated enzymes were detected in fasting animals, except for lipase. However, the intensities of the enzymatic activities were weaker in the fasting specimens relative to the fed specimens between days 7 and 10.     

Isolation and characterization of a fish F-type lectin from gilt head bream (Sparus aurata) serum

A novel fucose-binding lectin, designated SauFBP32, was purified by affinity chromatography on fucose–agarose, from the serum of the gilt head bream Sparus aurata. Electrophoretic mobility of the subunit revealed apparent molecular weights of 35 and 30 kDa under reducing and non-reducing conditions, respectively. Size exclusion analysis suggests that the native lectin is a monomer under the selected experimental conditions. Agglutinating activity towards rabbit erythrocytes was not significantly modified by addition of calcium or EDTA; activity was optimal at 37 °C, retained partial activity by treatment at 70 °C, and was fully inactivated at 90 °C. On western blot analysis, SauFBP showed intense cross-reactivity with antibodies specific for a sea bass (Dicentrarchus labrax) fucose-binding lectin. In addition, the similarity of the N-terminal sequence and a partial coding domain to teleost F-type lectins suggests that SauFBP32 is a member of this emerging family of lectins.     

Gonadogenesis in early developmental stages of Acipenser naccarii and influence of estrogen immersion on feminization

Gonad development processes and the effects of a single 8‐hour immersion treatment with 17β‐estradiol (E2, 400 μg L⁻¹) on sex differentation in the Adriatic sturgeon, Acipenser naccarii, were investigated. After migration of germ cells, gonadal ridges appeared in 16‐ to 18‐day old larvae and undifferentiated gonads in 55‐ to 60‐day old larvae. Putative ovaries with notches in the germinal epithelium and presumed testes with smooth germinal epithelium appeared in 180–185‐day old juveniles. Ovaries with proliferating oogonia and early meiotic oocytes clusters were observed in 292‐day old juveniles. Testes did not exhibit germ cell mitosis until 430 days of age. Developmental stages in E₂‐treated animals closely followed those of controls up to 430 days. The treatment significantly increased the percentage of ovaries when administered to embryos about 1.5 day before hatching, while did not significantly altered the normal 1/1 sex ratio when administered to 1.5‐day old pre‐larvae and 10‐day old larvae. It is likely that in A. naccarii exogenous E₂ administration may act through a feedback mechanism of self‐supporting steroid production and that steroids are the physiological inducers of sex differentiation, as in most teleosts. The E₂‐immersion treatment, easier than time‐consuming administration through food, could be a good approach to control sex differentiation and caviar production.