This study investigated the spatiotemporal distribution of abundances of the fish eggs and Engraulid eggs in relation to environmental factors along the coastal waters off southwestern Taiwan. Moreover, the key to fish eggs under an optical microscope in winter(January) was revised. Samplings were carried out on a bimonthly basis. In total, 38 tows were conducted from January to November 2003.The samplings of eggs were collected by horizontal hauls with NORPAC and cyclindricalconic net at the water surface along Jiading, Linyuan and Fangliao. Sea surface temperature, sea surface salinity, dissolve oxygen, chlorophyll a concentration and density were measured simultaneously with a seabird CTD. The spatiotemporal distribution of the total eggs abundances of two kinds of nets were very similar, and more the abundance of cyclindricalconic net than the NORPAC net. Therefore, the total eggs abundance and species of cyclindricalconic net were adopted to investigate the spatiotemporal distribution of the eggs along the coastal waters off southwestern Taiwan. The total abundances of fish eggs demonstrated significant seasonal variations highest in July and lowest in November. The geographical difference was also observed that the highest abundances were recorded at Jiading, and followed by Linyuan and Fangliao. Based on the results of correlation analyses, it was revealed that the spatiotemporal fluctuations in the total abundances of fish eggs were significant positive correlated to the monthly variation of precipitation. 2,110 fish eggs were obtained from six samples in January, 2003, including Ophichthidae, Clupeidae, Synodontidae, Scaridae, Callionymidae, Soleidae , Cynoglossidae and Engraulidae. The fish eggs that couldn¡¦t be identified were firstly sorted into 28 groups by the egg shape, size, and membrane pattern. Furthermore, they were stored into 64 species based on the presence or absence, and the pattern of pigmentation. The group BIII, Engraulids eggs and unknown dead eggs were the top three dominant species. Due to the different composition of the dominant species, there were significant differences in the spatial distribution. The species composition of dominant group were the Callionymids eggs and unknown dead eggs in Jiading. The species composition of dominance group were group BIII , Engraulids eggs and unknown dead eggs in Linyuan, and Fangliao. In 2003, Engraulids eggs were very abundant in the coastal waters off southwestern Taiwan, representing 14% of the total fish egg abundances. Two peaks of Engraulids eggs abundances were found. One in the July to September 2003, and other one in January 2003. Spatial distribution in the abundances were also observed that the highest abundances were recorded at Fangliao, and followed by Linyuan and Jiading. The mortality rate of the Engraulids eggs was the reverse trends and spatial distribution of the Engraulids eggs abundances. The abundances of eggs were increased when precipitation was high and abundance of Amphipoda was low. The shape of Engraulids eggs was displayed regional differences. Engraulids eggs in Jiading and Fangliao were elongated shape, and oval shape in Linyuan. Seasonal variation in the egg shape was observed and also segregated into two groups representing elongated eggs group in May, July and September and oval eggs group in January, March and November, respectively. Engraulids eggs longitudinal diameter was significantly positive correlated with the SST and monthly precipitation, and were negatively correlated with sea surface salinity, dissolve oxygen and density. Maybe the group BIII was Mugilids eggs. Their spawning ground occurred in Linyuan. Furthermore, the spawning ground of both Callionymids eggs and Synodontids eggs were at Linyuan and Fangliao. The main spawning grounds of Engraulids eggs were at Linyuan and Fangliao along the coastal waters off southwestern Taiwan. The peak spawning season were in summer and winter 2003. The spawning time of Engraulids eggs was approximately at 00:00 to 01:00. The spatiotemporal variations of Engraulids eggs size were influenced by the alternation of the spawning grounds of Engraulids fish along the coastal waters off southwestern Taiwan.

<p>Natural and constructed rocky reef habitats constitute important areas for lithophilic spawning fishes and their embryonic and larval offspring. Interstitial spaces created by the structure of rocky reefs create micro-environments where incubating embryos and juvenile fishes are potentially protected from predators. However, if interstitial spaces are filled or blocked by sediment deposition or biofouling, the reef structure may lose the protective benefits for embryonic and larval fish survival. Lake whitefish (<i>Coregonus clupeaformis</i>) and walleye (<i>Sander vitreus</i>) are native Great Lake lithophilic broadcast spawning fish that use rocky spawning habitats that are vulnerable to degradation caused by deposition of suspended sediments. To restore degraded rocky reef habitat, common practices include addition of material to existing reef structures or construction of new reefs, but both of these practices can be costly and time intensive. In this study, we measured the effect of different types and amounts of sediment cover on hatching success of walleye eggs and assessed if differences in female walleye (female length and egg size) account for tolerance to sediment cover. Additionally, we explored an alternative approach for reef restoration, custodial maintenance, in which we created two novel devices to potentially clean rocky reef habitat. We carried out two laboratory experiments in 2018 and 2019 to test the effect of sediment cover on hatching success of walleye eggs (2018) and to test how female identity and female length or egg size may interact with sediment cover to influence hatching success (2019). We exposed walleye eggs to instantaneous sediment cover (0 mm – 7mm) of either sand (course) or silt (fine) sediments from fertilization until day 15 of incubation. Our results indicated that walleye eggs were sensitive to silt cover (71% mortality- 2 mm cover silt) but not sand (47% mortality- 7mm cover sand). While there was an indication that hatching success was marginally related to female length and egg size, we concluded that sediment cover seemed to have similar effects on eggs, regardless of female length or egg size. The susceptibility of walleye eggs to mortality caused by sediment cover underscores the need for non-degraded spawning habitat. Our two cleaning devices used either propulsion or pressurized water jets to clean sediments from the rocky structure as they were towed behind a small vessel (i.e., did not require the use of SCUBA divers). We used devices to clean two natural rocky reefs in Saginaw Bay, Lake Huron in 2018 and 2019. We measured relative hardness before and after use of devices on cleaned and uncleaned study plots to determine effectiveness of devices. In addition, we measured egg deposition by fall (lake whitefish) and spring (walleye) lithophilic spawners on study plots to determine potential differences in fish usage of cleaned and uncleaned areas. We found that cleaning devices contributed to changes in relative hardness among study plots. Egg deposition was also variable on study plots but in general, egg deposition was consistently highest on treatment plots cleaned by our device that used propulsion. The practicality of cleaning devices was seemingly related to the magnitude of degradation of rocky reefs, nevertheless, our results show that the use of these or similar devices may potentially increase egg deposition by creating areas of higher-quality habitat. While more testing is necessary to fully understand the potential of our reef cleaning devices, this two-year study suggests that these devices may be capable of restoring degraded rocky spawning habitat which could potentially minimize the negative effects associated with sediment degradation on lithophilic spawning fish.</p>

Parmi les œufs de poisson, qui sont une ressource aquatique nutritionnelle intéressante, ceux de la truite arc-en-ciel (Oncorhynchus mykiss) contiennent une quantité élevée de protéines et une huile riche en acides gras polyinsaturés (AGPI), avec une proportion très importante de phospholipides. Cependant, l’œuf de poisson présente une capacité élevée d’auto-protection contre les contraintes extérieures, qui limite la destructuration de son réseau protéique par attaque enzymatique. Ainsi, le degré d’hydrolyse des œufs de la truite l’Alcalase®, la Neutrase® et la Protamex® varie entre 3 et 7 %, ce qui est très faible (20 % dans la majorité des protéines animales). L’extraction des lipides après protéolyse partielle est incomplète, probablement en raison d’interactions fortes avec les protéines faiblement hydrolysées. Ils contiennent une teneur élevée en phospholipides (53 % des lipides totaux) et les acides gras polyinsaturés entrent pour 42 % des acides gras totaux. Les AGPI, notamment le DHA, sont situés préférentiellement en position sn-2 sur la molécule de glycérol ce qui est particulièrement intéressant du point de vue nutritionnel. La stabilité à l’oxydation de l’huile a été étudiée par diverses méthodes, dont la spectrométrie infrarouge à transformée de Fourier. Cette méthode s’est avérée extrêmement intéressante pour une analyse structurale de la dégradation de l’huile en cours d’oxydation. Il peut être conclu que les lipides tirés des œufs de la truite arc-en-ciel ou de poisson en général, ont un réel avenir en matière de complément alimentaire ou nutraceutique, à condition de lever l’obstacle de l’hydrolyse enzymatique des protéines du chorion et du vitellus / Fish eggs, especially those of the rainbow trout (Oncorhynchus mykiss) in the present study, are an interesting nutritional aquatic source. They contain proteins of high value, as well as an oil rich in polyunsaturated fatty acids (PUFA) with a large percentage of phospholipids. However, they exhibit a high auto-protection capacity against environmental constraints and thus, the degree of hydrolysis of rainbow trout eggs by Alcalase®, Neutrase® and Protamex® proteases varied solely within 3-7 %. This value was low compared with the 20 % obtained in most animal proteins. The phospholipid content was high (53 % of total lipids) and PUFA accounted for 42 % of total fatty acids. Among PUFA, DHA was found preferably at the sn-2 position of the glycerol backbone, which is of special interest about nutritional properties. The oil release by enzymatic hydrolysis was found limited compared with chemical methods, probably because of the strong interactions engaged with the incomplete destructured protein network. The oxidative stability of the oil was studied through several methods in which the infrared Fourier transform appeared as the best tool for structural analysis along the oxidation process. As a conclusion, lipids from fish eggs, especially from rainbow trout, could be a nutritional breakthrough, as far the enzymatic hydrolysis of the vitellus and of the chorion proteins is achieved

In marine teleosts that spawn pelagic eggs (pelagophils), the process of oocyte hydration that occurs during meiosis resumption is a key physiological process for the survival of the eggs in the ocean. Previous studies have discovered the role of a teleost-specific aquaporin water channel (Aqp1b) during fish oocyte hydration, but direct experimental evidence for the function of Aqp1b in oocytes is still lacking. In addition, the molecular regulation of the Aqp1b-mediated mechanism remains poorly understood. In this context, the main objectives of the present thesis were to investigate the evolutionary origin of aqp1ab in teleosts, to provide functional evidence of the role of Aqp1b during oocyte hydration, and to begin to dissect the molecular mechanisms involved in the transcriptional regulation of aqp1b in the oocyte of marine teleosts. By integrating the molecular phylogeny with synteny and structural analyses we show that the teleost aqp1aa and -1ab paralogs (previously annotated as aqp1a and -1b, respectively) arose by tandem duplication, and that the Aqp1ab C-terminus is the most rapidly evolving subdomain within the vertebrate aquaporin superfamily. The functional role of Aqp1ab was investigated in Atlantic halibut (Hippoglossus hippoglossus), a marine acanthomorph teleost that spawns one of the largest pelagic eggs known. Using immunological inhibition of Aqp1ab in halibut oocytes and artificial expression of the halibut paralog Aqp1aa, we demonstrate that Aqp1ab is required for full hydration of oocytes undergoing meiotic maturation. To investigate the aqp1ab transcriptional regulation in oocytes, we isolated the 5’-flanking region of the gilthead seabream (Sparus aurata) aqp1ab gene which contains regulatory cis-elements for the nuclear progestin receptor (Pgr) and SOX transcription factors. The Pgr, as well as sox3 and -8b transcripts, are co-expressed in seabream oo-gonia, whereas in primary growth oocytes, when aqp1ab mRNA and protein are synthesized, the Pgr is translocated into the nucleus. In contrast, sox9b is highly expressed in more advanced oocytes showing the depletion of aqp1ab transcripts. In the seabream, four different pgr transcript variants are expressed in primary growth ovaries which are generated by alternative pre-mRNA splicing. Seabream wild-type Pgr shows the highest transactivation response to progestins such as 17α,20β-dihydroxy-4-pregnen-3-one (17,20β-P) and 17α,20β,21-trihydroxy-4-pregnen-3-one (17,20β,21-P), whereas two of the N-terminally truncated Pgr isoforms regulate novel nuclear and cytosolic mechanisms of dominant-negative repression of Pgr-mediated transcription. Transactivation assays on the aqp1ab promoter demonstrated that aqp1ab transcription is dependent on wild-type Pgr, with Sox3 and -8b acting synergistically, while Sox9b acts as a repressor. Incubation of primary ovarian explants in vitro with 17,20β-P, followed by chromatin immunoprecipitation, confirmed that 17,20β-P-activated Pgr enhanced aqp1ab promoter activation. The production of 17,20β-P in seabream primary growth ovaries in vivo was consistent with the expression of P450c17-II (Cyp17a2) and 20β-hydroxysteroid dehydrogenase (Cbr1), enzymes needed for progestins synthesis, in granulosa cells associated with primary growth oocytes, and with a high concentration of 17,20β-P. Incubation of primary ovarian explants with recombinant piscine follicle-stimulating hormone (rFsh) in vitro stimulated 17,20β-P synthesis, which was reduced in the presence of Cbr1 inhibitors. The rFsh-mediated production of 17,20β-P correlated with the up-regulation of cyp17a2 and cbr1 transcription, as well as of wild-type pgr mRNA and protein levels. Altogether, these data suggest that aqp1ab transcription in seabream primary growth oocytes is under Fsh regulation through the synthesis of progestins. The results of this thesis show that the Aqp1ab mediated mechanism for oocyte hy-dration is likely conserved in marine teleosts. In addition, the tight transcriptional reg-ulation of Aqp1ab during oogenesis highlights the essential physiological role of this water channel and opens new research avenues for understanding the molecular basis of egg formation in marine fish. / La hidratación de los oocitos de teleósteos marinos que producen huevos pelágicos es clave para la supervivencia de los embriones en el océano. Estudios previos han descu-bierto el papel de una acuaporina específica de teleósteos (Aqp1b) durante este proceso, pero se carece todavía de evidencias experimentales directas, así como información sobre la regulación molecular de la Aqp1ab. En la presente tesis, estudios iniciales con el fletan Atlántico (Hippoglossus hippoglossus) confirman que los parálogos aqp1aa y -1ab de teleósteos han surgido probablemente por duplicación génica en tándem, y han demostrado el papel esencial de la Aqp1ab durante la hidratación del oocito. Para investigar el control transcripcional del gen aqp1ab en los oocitos de la dorada (Sparus aurata) se ha aislado su región promotora, la cual contiene elementos cis-reguladores de unión al receptor nuclear de progestinas (Pgr), como la 17α,20β-dihidroxi-4-pregnen-3-ona (17,20β-P), y factores de transcripción Sox. Estudios de localización subcelular indican que el Pgr aparece en el citoplasma de las oogonias, así como en el núcleo de oocitos en crecimiento primario (pre-vitelogénicos) coincidiendo con la activación de la expresión de aqp1ab. En este estadio también se expresan cuatro isoformas diferentes del Pgr, dos de las cuales pueden inhibir la transcripción mediada por el Pgr de forma dominante negativa. Las oogonias también expresan sox3 y -8b, mientras que el sox9b aparece en el estadio de alveolo cortical, cuando se reduce la expresión de aqp1ab. Ex-perimentos de transactivación indican que el Pgr activa la transcripción de aqp1ab, con Sox3 y -8b actuando de forma sinérgica, mientras que el Sox9b reprime este mecanismo. El papel del Pgr se ha investigado sobre explantes ováricos pre-vitelogénicos incubados in vitro, lo cual ha demostrado que la 17,20β-P, producida en las células de la granulosa en respuesta a la hormona folículo estimulante, activa el promotor de aqp1ab en el oocito induciendo la síntesis de Aqp1ab. Los resultados de esta tesis revelan por primera vez una estricta regulación transcripcional del gen aqp1ab durante la oogénesis de teleósteos marinos, lo cual remarca la función fisiológica esencial de este canal de agua durante la formación de los huevos pelágicos.