Effect of rearing technique over skeletal deformities incidence and osteological development in red porgy Pagrus pagrus (Linnaeus, 1758) larvae

[EN]Red porgy is a candidate species for marine aquaculture diversification.The objective of the present study was to describe the osteological development and the occurrence of skeletal deformities in Pagrus pagrus larvae in relation to the intensification of the rearing system. Fish samples were periodically collected along the development from hatching to juveniles (95days after hatching). Osteological development and the presence of skeleton abnormalities were evaluated. X-ray studies revealed a high number of fish (Semi-intensive:38.8%; Intensive:46.5 %) with skeletal deformities. No significant interaction was found on the incidence of lordosis and fused vertebrae with the rearing tecnique. However, cranial abnormalities and kyphosis incidences were significantly higher in intensive system cultured red porgy. Also, the position of fused vertebrae in this fish was located mainly in the caudal area instead of pre-hemal area for semi-intensive system reared red porgy. Present results, suggest a relationship among feeding sequence, osteological development and deformity incidence and location in red porgy larvae

Sea bream (Sparus aurata) intensive larval rearing : effects of microalgae and phages addition on the performance of the larvae

Máster Oficial en Cultivos Marinos. Trabajo presentado como requisito parcial para la obtención del Título de Máster Oficial en Cultivos Marinos, otorgado por la Universidad de Las Palmas de Gran Canaria (ULPGC), el Instituto Canario de Ciencias Marinas (ICCM), y el Centro Internacional de Altos Estudios Agronómicos Mediterráneos de Zaragoza (CIHEAM)

Physiological and metabolic effects of nutrient enrichment on adult Artemia franciscana

[EN] Brine shrimp nauplii (Artemia sp.) are used in aquaculture as the major food source for many cultured marine larvae, and also used in the adult phase for many juvenile and adult fish. One artemia species, Artemia franciscana is most commonly preferred, due to the availability of its cysts and to its ease in hatching and biomass production. The problem with A. franciscana is that its nutritional quality is relatively poor in essential fatty acids, so that it is common practice to enrich it with emulsions like SELCO and ORIGO. This “bioencapsulation”, enrichment method permits the incorporation of different kinds of products into the artemia. This brine-shrimp’s non-selective particle-feeding habits, makes it particularly suitable for this enrichment process. The bioencapsulation is done just prior to feeding the artemia to a predator organism. This allows the delivery of different substances, not only for nutrient enrichment, but also for changing pigmentation and administering medicine. This is especially useful in culturing ornamental seahorses and tropical fish in marine aquaria In this study the objectives were to determine, the relative nutrient value of ORIGO and SELCO as well as the optimal exposure to these supplements prior to their use as food-organisms.

Effect of different live prey, Grapsus grapsus Linnaeus 1758 and Plagusia depressa Fabricius, 1775 zoeas,over histology and biochemical composition of common octopus, octopus vulgaris Cuvier, 1797 paralarvae

[EN] Since paralarval rearing is still the main bottleneck for the development of octopus culture, the aim of the present study was to obtain some information on the feeding strategy and nutritional requirements during paralarval stage. For that purpose just hatched out octopus paralarvae were fed with live preys in three different combinations, trying to match their natural food: Enriched Artemia metanauplii, Grapsus grapsus zoeas supplemented with enriched Artemia, and Plagusia depressa zoeas supplemented with enriched Artemia. Paralarval treatments were carried out during 28 days in triplicates; fibre glass 120 l tanks in flow through system were used. Growth, in terms of dry body weight, mantle length and width, was determined each seven days. A histological study of the paralarval development was carried out. Biochemical composition of preys and paralarvae were determined. Growth was significantly better in paralarvae fed with zoeas and Artemia than in those fed only with Artemia, from day 8 after hatching. Besides a clear effect on the digestive gland histology morphology was observed.

Effect of culture system and diet in early larval stages over skeletal deformities development in gilthead sea bream

[ES] Main deformities such as lordosis, opercular deformities and upper/lower jaws shortening are considered as quality descriptors in commercial marine fish fry production and seem to be related with larval culture conditions in early larval stages. The aim of this work was to obtain information about the contribution of the diet and rearing system to the apparition of these abnormalities in gilthead sea bream (Sparus aurata) larvae in semi-industrial scale facilities. For that purpose, two different larval rearing systems semi-intensive and intensive were compared; besides, two different rotifer enrichments, DHA Protein Selco, (Inve Aquaculture, Dendermonde, Belgium) (R1) and Red Pepper Paste, (Bernaqua bvba, Turnhout, Belgium) were tested in the intensive system. Biochemical composition of larvae, preys and commercial products was analysed. At 50 days post hatching six hundred fish per treatment were individually studied under stereoscope and deformity frequency recorded. Besides at 95 days post hatching fry were soft X ray monitored. Both rotifer enrichment and rearing system affected survival, growth and deformity frequency. Rearing system did not affect total larvae fatty acid content except at 20 dah, where DHA were significantly higher and EPA significantly lower in Semi-intensive system. A significantly lower percentage of deformity rates together with better survival and growth were obtained in the semi-intensive system. In dietary treatment, rotifer enrichment significantly affected larval survival. R1 rotifers enrichment significantly (P<0.05) improved survival when compared to fed R2 larvae. The content of DPA was significantly (P<0.05) higher in R2 fed larvae reflecting the R2 rotifers content of this fatty acid. The level of this FA tended to decrease in concordance with the rotifers replacement by artemia in the diet. The effects n-3-HUFA and DPA (22:5n-6) over larval survival and skeletal deformities development is discussed.

Effect of rearing techniques over, survival growth and skeletal abnormalities development in red porgy (Pagrus Pagrus) larvae

[EN] Red porgy is one of the most interested new species for Spanish and other Mediterranean countries. Although no industrial procedures for fry production has been yet developed. The aim of this work was to develop an industrial scale larval rearing protocols testing the viability of two different rearing techniques (semi-intensive vs intensive) in pilot scale facilities. The second objective was to obtain information about the contribution of rearing system to the apparition of morphological abnormalities such as lordosis, opercular deformities and upper/lower jaws shortening which are considered as quality descriptors in commercial marine fish fry production and seem to be related with larval culture conditions in early larval stages. For that purpose, two different larval rearing systems semi-intensive and intensive were compared using the same live feed enrichments. Biochemical composition of larvae, preys and commercial products was analysed. At 50 days post hatching six hundred fish per treatment was individually studied under stereoscope and abnormalities frequency recorded. At 95 days post hatching fry were soft X ray monitored as well. Survival and abnormalities frequency were similar between treatments although a better growth in terms of total length was obtained in the semi-intensive system.

Development of skeletal deformities in Gilthead sea bream (Sparus aurata) reared under different larval culture and dietary conditions

[EN] Main deformities such as lordosis, opercular deformities and upper/lower jaws shortening are considered as quality descriptors in commercial marine fish fry production and seem to be related at least with larval culture conditions in early larval stages. The aim of this work was to obtain information about the contribution of the diet and rearing system to the apparition of these abnormalities in gilthead sea bream (Sparus aurata) larvae in semi-industrial scale facilities. For that purpose, two different larval rearing systems semi-intensive and intensive were compared by duplicate and with the same live feed enrichments; besides, two different rotifer enrichments were tested in an intensive system. Biochemical composition of larvae, preys and commercial products was analysed. At 50 days post hatching six hundred fish per treatment were individually studied under stereoscope and abnormalities frequency recorded. At 95 days post hatching fry were soft X ray monitored as well. Survival and malformation frequency were significantly different between treatments, the effect of diet and system are discussed. A significantly lower percentage of deformity rates together with better survival and growth were obtained in the semi-intensive system, whereas the rotifer enrichment significantly affected larval survival.

Crecimiento y supervivencia de paralarvas de O. vulgaris cultivadas bajo diferentes intensidades de luz.

[EN] Paralarval rearing constitutes the main bottleneck to the complete development of octopus culture. This paper try to optimise the larval rearing conditions of Octopus Paralarvae, testing the effect of light intensity (1000, 3000 and 6000 lux ) over growth and paralarval survival. Four fibber glass tanks, 100 litter capacity with an initial density of 20 ind./litter were used for each treatment. All light treatments were fed with the same protocol, just hatching Maja Squinado Zoeas and Artemia metanaulios (72 hours)enriched with a commercial emulsion (Red Pepper Paste, Bernaqua, bva; Belgium). Better growth and survival were found with the highest light intensity treatment.

Comparación entre la calidad de puestas naturales e inducidas, mediante inyección e implante de GnRHa, del pez de limón (Seriola dumerili).

[EN]For this study, a total of 19 greater amberjack (Seriola dumerili) broodstock were used. Females with oocites bigger than 600μ, and fluent males with a mean weight of 10.72±1.22 kg, fameles, and 10.77±2.33 kg males. The experimental tanks were circular tanks of 40 m3 capacity. One of the tanks were not induced and the other two were with GnRHa, one with injections and the other by implants. Best results, in all parameters studied were with natural spawns, with a 84.37% of fertilization, 92.21% of viable eggs at 24 hours of spawn, 96.60% of hatching, 69.91% of larvae one day after hatching and 10.78% of 5 day life larvae.

Efecto del tipo de hormona y del metodo de aplicación, en las producciones de reproductores de corvina (Argyrosomus regius).

[EN]In the present study, the effect of three different hormonal treatments were tested (GnRHA and hCG injected and GnRHa implanted), in meagre (Argyrosomus regius) broodstock born in captivity, and their production. The highest productions per kg of female and spawn: number of eggs, fertilized eggs, hatched larvae, and larvae 3 days old after hatching, corresponds to the treatment of GnRHa injected. The number of viable eggs, hatched eggs and larvae 3 days with GnRha injected was significantly different from those treated with hGC. No significant differences were observed in production per kg female and induction in the different hormonal treatments,

Inducción de reproductores de corvina (Argyrosomus regius) nacidos en cautividad, mediante inyección con GCH y con GnRHa, esta aplicada además mediante implante. Efecto sobre la la calidad de las puestas

[EN]The best results in all quality parameters, were obtained in the spawns of the broodstock injected with GnRHa, having 98,7 + 1,4% of fertilized eggs, 62,24 ± 27,47 % viable eggs, 92,56 ± 17,19 % hatching and 92,73 ± 10,52 % 3 day old larvae after hatching. Percentage of viable eggs was lower in the spawns of the broomstick injected with hCG, and had significant differences when compared with GnRHa broodstock treatments. 3 day old larvae and hatchings, obtained from the broomstock injected with GnRHa showed significant differences to the rest of the hormonal treatments.

Morphological, histological, histochemical and behavioral aspects during early development of red porgy Pagrus pagrus L. reared in mesocosm.

[EN]The present study provides a description of the morphological, histological and histochemical development, and the ecological implications of red porgy larvae reared in mesocosm, from hatching to 30 days after hatching (DAH).

Nuove tecniche per l'allevamento massale di Ditteri del genere Coenosia Meigen

The species of the genus Coenosia Meigen are polyphagous predators both in the larval and in the adult stage. In Europe five Coenosia species of the ‘tigrina group’ are naturally present in greenhouses, where they can establish for long periods. As their wide preys range includes important pests of protected crops, as Aleyrodidae, Sciaridae and Agromyzidae, Coenosia species are considered promising potential biological control agents. A method for rearing Coenosia species in vivo was developed for the first time in 1993 in Germany, where C. attenuata, C. strigipes and C. humilis were bred on Bradysia paupera (Diptera Sciaridae), reared on Fusarium spp. cultivated on wood fibre. Although this method was partially simplified afterwards, it is still too complex and expensive for a mass production. This research aimed at simplifying this rearing procedure and making it cheaper, in the perspective of an eventual mass production. Studies on potential preys were conducted, to determine their suitability for C. attenuata larvae and adults and to develop rearing methods. Biology and rearing methods of Bradysia paupera Tuomikoski, Scatella stagnalis Fallén and Drosophila melanogaster Meigen (Diptera: Sciaridae, Ephydridae, Drosophilidae) were compared. B. paupera resulted the most suitable prey for rearing C. attenuata in vivo. The Sciarid fly was effectively reared on damp coconut fibre with fresh Agaricus bisporus (J.E. Lange) Pilát, thus simplifying the existing method. After preliminary trials with different potential preys, attempts to rear C. attenuata in vivo on B. paupera and D. melanogaster were made. The best results were obtained with B. paupera, reared on coconut fibre and A. bisporus, but the method needs further improvement. Trials of in vitro rearing of C. attenuata were also made: as no specific diet for Coenosia species is reported in literature, different potentially suitable media were tested. Among these, a specific diet for Diptera Tachinidae resulted a good starting point for further studies and improvements. The biology of C. attenuata adults captured in greenhouses was also studied, by observing both groups and isolated individuals. Data on lifespan, daily number of preys per adult, daily number of laid eggs and hatching rate were recorded, and the effects of different foods on these parameters were analyzed. The following foods were compared: D. melanogaster adults only, as preys for C. attenuata; D. melanogaster adults and a water-honey solution; the water-honey solution only. Honey resulted an effective food integration for C. attenuata, increasing lifespan and the number of egg laying females. It is possible that in greenhouses Coenosia adults complete the preys diet with nectar and/or honeydew. Moreover, the integration with honey reduced the daily preys consumption. This may allow to prevent cannibalism among Coenosia adults in the rearing conditions, where high population densities are required. A survey of the Coenosia species naturally present in Lombardy greenhouses was conducted. The species C. attenuata, C. strigipes, C. tigrina and C. atra were detected. C. attenuata resulted the most common, recorded in most greenhouses and for consecutive years. Besides, the presence of potential preys, weeds and the crops were recorded in each greenhouse. Nevertheless, it is difficult to determine the relation between these parameters and the presence of Coenosia species.

L’allevamento della sogliola comune (Solea solea L.): riproduzione controllata e nuovi sviluppi di nutrizione e alimentazione in fase larvale e giovanile

In questa tesi sono illustrate alcune sperimentazioni finalizzate alla standardizzazione del ciclo produttivo della sogliola comune (Solea solea) in cattività. E’ stato creato un parco di riproduttori selvatici ed è stata standardizzata la riproduzione ad un livello compatibile con la realtà produttiva del settore. Indagini genetiche di assegnazione parentale hanno evidenziato come alcuni esemplari siano stati predominanti negli accoppiamenti e nel conseguente contributo alla generazione della prole. Ciò ha determinato una diminuzione della variabilità genetica dei discendenti. La composizione quali-quantitativa degli acidi grassi delle uova è stata correlata con la sopravvivenza larvale nel corso di un’intera stagione riproduttiva. Tale composizione non ha subito importanti variazioni su scala temporale e sembra essere stata influenzata dall’alimentazione somministrata ai riproduttori nel periodo precedente alla riproduzione. Le analisi di interazione tra momento riproduttivo e qualità delle uova hanno confermato che è stato possibile ottenere uova di buona qualità in termini di sopravvivenza larvale nel corso di tutta la stagione riproduttiva. Larve di sogliola sono state svezzate precocemente 13 giorni dopo la schiusa riducendo l’impiego di cibo vivo a favore di micro diete commerciali. Tale svezzamento ha ridotto le performance di accrescimento, ma non la sopravvivenza e lo sviluppo della metamorfosi quando comparati ad un trattamento standard. La riduzione del cibo vivo ha ottimizzato i costi di produzione e migliorato l’igiene in vasca. L’ontogenesi di precursori di enzimi digestivi è stata determinata tramite PCR quantitativa. I risultati di espressione di tripsinogeno, chimotripsinogeno e amilasi hanno mostrato come tali enzimi rivestano un ruolo chiave nei processi digestivi delle prime fasi larvali. Esemplari giovanili hanno ottenuto un significativo maggiore indice di accrescimento e migliore indice di conversione quando alimentati con diete sperimentali contenenti un elevato tenore proteico. Un aumento dell’incidenza di vacuoli lipidici a livello epatico è stato osservato all’aumentare del tenore proteico della dieta.

Funktionelle und strukturelle Charakterisierung des Ovastacins in vivo und in vitro

Die Metalloprotease Ovastacin, ein Vertreter der Astacin-Familie, wurde erstmals 2004 beschrieben. Im Ovar von Säugetieren ist Ovastacin-mRNA im Zeitfenster vom Stadium der Sekundärfollikel bis kurz nach der Befruchtung der Eizelle zu finden. Der Expressionsort und -zeitpunkt sowie die Sequenzähnlichkeit von über 60% mit sogenannten „Schlüpfenzymen“ (engl. hatching enzymes), die man in den Eizellen und Zygoten niederer Wirbeltiere und Wirbelloser gefunden hatte, ließen die Vermutung aufkommen, es könnte sich hier um das Säugerhomolog dieser Proteasen handeln. Generell lösen hatching Enzyme die derben embryonalen Hüllstrukturen (bei Säugern die Zona pellucida, ZP) beim Schlüpfvorgang auf. Die essentielle Bedeutung des Ovastacins für die Befruchtung wird durch die um ca. 30% reduzierte Fruchtbarkeit von Ovastacin defizienten Mäusen belegt. Hochinteressant war in diesem Zusammenhang die Entdeckung des Ovastacins in den Cortikalgranula der Oocyten sowie seine Fähigkeit, das Zona pellucida Protein 2 zu schneiden. Die dadurch bewirkte Verhärtung der Zona pellucida verhindert das Eindringen weiterer Spermien, das heißt sie baut eine Barriere gegen Polyspermie auf. Ziel dieser Arbeit war es, Belege für die physiologische Funktion des Ovastacins zu finden. Vor allem galt es, potentielle Aktivatoren zu identifizieren, da das Enzym wie alle Astacine als inaktive Vorstufe gebildet wird, die proteolytisch aktiviert werden muss. Zu diesem Zweck exprimierte ich rekombinantes Pro-Ovastacin in Insektenzellen. Aktivierungsstudien in vitro zeigten, dass ein saures Milieu zu einer Aktivierung führt, ohne die Abspaltung des Propeptids zu bewirken. Sequenzalignments und ein homologes Strukturmodell des Ovastacins wiesen auf Trypsin- oder Elastase-ähnliche Serinproteasen als potentielle Aktivierungsenzyme hin. Tatsächlich konnte mit diesen beiden Proteasetypen zum ersten Mal aktives Ovastacin aus Pro-Ovastacin erzeugt werden. Trypsin kommt als physiologischer Aktivator allerdings nicht in Betracht, da es bisher in keinem der Gewebe nachgewiesen werden konnte, in dem Ovastacin exprimiert wird. Die neutrophile Elastase dagegen konnte in der Leber, im Herz sowie im Blutplasma nachgewiesen werden. Mit Hilfe spezifischer Antikörper konnte das Herz als Expressionsort für Ovastacin bestätigt werden. Somit wäre Elastase ein potentieller physiologischer Aktivator von Ovastacin. Die Identifikation des Ovastacins in Geweben wie Leber, Herz, Nabelschnur und im Blutplasma weist auf eine Rolle der Protease in proteolytischen Netzwerken außerhalb der Spermien-Ei-Interaktion hin. Die Bedeutung der biologischen Kontrolle des Ovastacins bei der Befruchtung der Säugereizelle wird durch die Beobachtung untermauert, dass das Leberprotein Fetuin B als physiologischer Ovastacininhibitor fungiert und dadurch eine vorzeitige Verhärtung der Zona pellucida verhindert, die andernfalls die Penetration von Spermien prinzipiell verhindern würde.