Dissertations / Theses on the topic 'Sarbach'

Questa tesi si occupa della questione della stabilità lineare di wormholes (tunnel spaziotemporali) statici e a simmetria sferica, supportati da campi scalari di tipo fantasma autointeragenti, nel contesto della Relatività Generale per spazitempi di dimensione arbitraria. In letteratura, attraverso un'analisi gauge-invariante delle configurazioni di tipo wormhole, spesso si riesce a disaccoppiare le equazioni di campo linearizzate, ottenendo un'equazione delle onde (master equation) che, tuttavia, tipicamente è singolare dove il coefficiente radiale della metrica ha un punto critico, cioè nella gola del tunnel. Per risolvere questo problema, nei lavori passati è stato proposto un metodo di regolarizzazione che trasforma l'equazione delle onde singolare in una regolare; questo metodo è solitamente denominato "S-deformazione" (e spesso richiede parzialmente un'implementazione numerica, specialmente nel caso di campi scalari con un'autointerazione non banale). Il primo risultato del mio lavoro è la riduzione delle equazioni di campo linearizzate ad un sistema delle onde vincolato e completamente regolare, per due funzioni gauge-invarianti delle perturbazioni dei coefficienti della metrica e del campo scalare, opportunamente definite; il secondo risultato è una strategia per disaccoppiare questo sistema, ottenendo una sola master equation delle onde per un'altra quantità gauge-invariante. Nessun passaggio di questa costruzione determina l'apparizione di singolarità nella gola del tunnel o in altri punti (sempre che il campo scalare imperturbato non abbia punti critici, cosa che accade in moti esempi); quindi non è necessario regolarizzare a posteriori la master equation utilizzando il metodo di S-deformazione. Questo formalismo gauge-invariante e libero da singolarità, che generalizza a dimensione arbitraria l'approccio del mio articolo [1], è applicato ad alcune soluzioni di tipo wormhole statiche note (la maggior parte, ma non tutte, considerate in [1]). La più importante applicazione è ad un wormhole Anti-de Sitter (AdS), la cui stabilità lineare non pare sia mai stata analizzata da altri autori finora; utilizzando il presente metodo è possibile derivare una master equation completamente regolare che descrive le perturbazioni del wormhole AdS e quindi dimostrare che quest'ultimo è linearmente instabile, dopo aver dettagliatamente analizzato le proprietà spettrali di un operatore di tipo Schrödinger che compare nella master equation. Sulla stessa linea, è ottenuto un risultato parziale per l'analogo wormhole di tipo de Sitter (dS), caso tecnicamente più sottile a causa della presenza di orizzonti. Come ulteriore applicazione, ho riottenuto in maniera libera da singolarità le master equations per le perturbazioni di dei wormholes di Ellis-Bronnikov e di Torii-Shinkai. Ad integrazione, l'instabilità lineare dei wormholes AdS e di Torii-Shinkai sono riottenute utilizzando un metodo alternativo, privo di singolarità ma gauge-dipendente: in questo caso, si ottiene una master equation per la perturbazione della coordinata radiale, e l'indipendenza dal gauge del risultato di instabilità è testata a posteriori. Questo approccio alternativo e gauge-dipendente generalizza quello introdotto in [2] per il wormhole di Ellis-Bronnikov a simmetria riflessiva. Vorrei citare infine [3], dal quale ho riportato alcuni fatti sui wormholes appena menzionati in assenza di perturbazione. BIBLIOGRAFIA: [1] F. Cremona, L. Pizzocchero, and O. Sarbach. Gauge-invariant spherical linear perturbations of wormholes in einstein gravity minimally coupled to a self-interacting phantom scalar field. Physical Review D, 101, 05 2020. [2] F. Cremona, F. Pirotta, and L. Pizzocchero. On the linear instability of the Ellis-Bronnikov-Morris-Thorne wormhole. Gen. Relativ. Gravitat., 51:19, 2019. [3] F. Cremona. Geodesic structure and linear instability of some wormholes. Proceeding for the conference: Domoschool 2019 (submitted).
In this thesis I deal with the linear stability analysis of static, spherically symmetric wormholes supported by phantom self-interacting scalar fields, in the framework of General Relativity with arbitrary spacetime dimension. In the previous literature, a gauge-invariant stability analysis of wormhole configurations often succeeds in decoupling the linearized field equations, yielding a wave-type master equation which, however, is typically singular where the radial coefficient of the metric has a critical point, that is, at the wormhole throat. In order to overcome this problem a regularization method has been proposed in previous works, which transforms the singular wave equation to a regular one; this method is usually referred to as “S-deformation” (and sometimes requires a partly numerical implementation, especially, in the case of scalar fields with nontrivial self-interaction). The first result of my work is the reduction of the linearized field equations to a completely regular, constrained wave system for two suitably defined gauge-invariant functions of the perturbations in the metric coefficients and in the scalar field; the second result is a strategy for decoupling this system, obtaining a single wave-type master equation for another gauge-invariant quantity. No step of this construction causes the appearing of singularities at the wormhole throat or elsewhere (provided that the unperturbed scalar field has no critical points, which occurs in many examples); therefore, it is not necessary to regularize a posteriori the master equation via the S-deformation method. This gauge-invariant and singularity-free formalism, which generalizes to arbitrary spacetime dimensions the approach of my paper [1], is then applied to some known static wormhole solutions (most, but not all of them considered in [1]). The most relevant application is a certain Anti-de Sitter (AdS) wormhole, whose linear stability analysis does not seem to have been performed previously by other authors; by using the present method, it is possible to derive a completely regular master equation describing the perturbations of the AdS wormhole and prove that the latter is actually linearly unstable, after providing a detailed analysis of the spectral properties of the Schrödinger type operator appearing in the master equation. A partial instability result is derived along the same lines for the analogous de Sitter (dS) wormhole, a technically more subtle case due to the presence of horizons. As a further application, I rederive in a singularity-free fashion the master equations for the perturbed Ellis-Bronnikov and Torii-Shinkai wormholes. As a supplement, the linear instability results for the AdS and for the Torii-Shinkai wormholes are also recovered using an alternative, singularity free but gauge-dependent method: in this case a regular master equation is derived for the perturbed radial coordinate, and the gauge-independence of the instability result is tested a posteriori. This alternative, gauge-dependent approach generalizes that introduced in my paper [2] for the reflection symmetric Ellis-Bronnikov wormhole. Let me also cite [3], from which I report some facts about the previously mentioned wormholes in absence of perturbations. BIBLIOGRAPHY: [1] F. Cremona, L. Pizzocchero, and O. Sarbach. Gauge-invariant spherical linear perturbations of wormholes in einstein gravity minimally coupled to a self-interacting phantom scalar field. Physical Review D, 101, 05 2020. [2] F. Cremona, F. Pirotta, and L. Pizzocchero. On the linear instability of the Ellis-Bronnikov-Morris-Thorne wormhole. Gen. Relativ. Gravitat., 51:19, 2019. [3] F. Cremona. Geodesic structure and linear instability of some wormholes. Proceeding for the conference: Domoschool 2019 (submitted).

Various aspects of the biology of the tarwhine Rhabdosargus sarba and western yellowfin bream Acanthopagrus latus were studied. The studies on R. sarba have focused on populations in temperate coastal marine waters at ca 32 degrees S and the lower reaches of an estuary (Swan River Estuary) located at the same latitude and in a subtropical embayment (Shark Bay) at ca 26 degrees S, while those on A. latus were conducted on the population in the latter embayment. A combination of a macroscopic and histological examination of the gonads demonstrated that R. sarba is typically a rudimentary hermaphrodite in Western Australian waters, i.e. the juveniles develop into either a male or female in which the ovarian and testicular zones of the gonads, respectively, are macroscopically undetectable. This contrasts with the situation in the waters off Hong Kong and South Africa, in which R. sarba is reported to be a protandrous hermaphrodite. However, it is possible that a few of the fish that are above the size at first maturity and possess, during the spawning period, ovotestes with relatively substantial amounts of both mature testicular and immature ovarian tissue, could function as males early in adult life and then change to females. Although R. sarba spawns at some time between late winter and late spring in Western Australia, spawning peaks later in the Swan River Estuary than in coastal, marine waters at the same latitude and Shark Bay, in which salinities are always close to or above that of full strength sea water, i.e. 35 0/00. While the males and females attain sexual maturity at very similar lengths in the Swan River Estuary and Shark Bay, i.e. L50s all between 170 and 177 mm, they typically reach maturity at an earlier age in the former environment, i.e. 2 vs 3 years old. Thus, length and consequently growth rate influence the timing of maturity rather than age. During the spawning period, only 9 % of the fish caught between 180 and 260 mm in nearshore, shallow marine waters had become mature, whereas 91 % of those in this length range over reefs were mature, indicating that R. sarba tends to move offshore only when it has become physiologically ready to mature. The L50s at first maturity indicate that the current minimum legal length in Western Australia (230 mm) is appropriate for managing this species. Oocyte diameter frequency distributions, stages in oocyte development, duration of oocyte hydration and time of formation of post-ovulatory follicles in mature ovaries of Rhabdosargus sarba in the lower Swan River Estuary (32 degrees 03'S, 115 degrees 44'E) were used, in conjunction with data on tidal cycles, to elucidate specific aspects of the reproductive biology of this sparid in an estuarine environment. The results demonstrated the following. (i) Rhabdosargus sarba has indeterminate fecundity sensu Hunter et al. (1985). (ii) Oocyte hydration commences at about dusk (18:30 h) and is completed by ca 01:30-04:30 h, at which time ovulation, as revealed by the presence of hydrated oocytes in the ovarian duct and appearance of newlyformed post-ovulatory follicles, commences. (iii) The prevalence of spawning was positively correlated with tidal strength and was greatest on days when the tide changed from flood to ebb at ca 06:00 h, i.e. approximately when spawning ceases. Spawning just prior to strong ebb tides would lead to the transport of eggs out of the estuary and thus into salinities that remain at ca 35 0/00. The likelihood of eggs being transported downstream is further enhanced by R. sarba spawning in deeper waters in the estuary, where the flow is greatest. (iv) Although mature ovaries were found in R. sarba in the estuary between early July and December, the prevalence of atretic oocytes was high until September, when salinities started rising markedly from their winter minima. Batch fecundities ranged from 2,416 for a 188 mm fish to 53,707 for a 266 mm fish. The average daily prevalence of spawning amongst mature females during the spawning period of R. sarba caught in the lower estuary, i.e. July to end of October, was 36.5 %. Thus, individual female R. sarba spawned, on average, at intervals of ca 2.7 days in each spawning season. Female R. sarba with total lengths of 200, 250 and 300 mm were estimated to have a batch fecundity of 7,400, 20,100 and 54,800 eggs, respectively and annual fecundities of 332,000, 903,000 and 2,461,000 eggs, respectively. Rhabdosargus sarba is shown to undergo size-related movements in each of the three very different environments in which it was studied. In temperate coastal waters, R. sarba settles in unvegetated nearshore areas and then moves progressively firstly to nearby seagrass beds and then to exposed unvegetated nearshore areas and finally to areas around reefs where spawning occurs. Although R. sarba spawns in the lower Swan River Estuary, relatively few of its early 0+ recruits remain in the estuary and substantial numbers of this species do not start reappearing in the estuary until they are ca 140 mm. In Shark Bay, R. sarba uses nearshore mangroves as a nursery area and later moves into areas around reefs. The maximum ages recorded for R. sarba in coastal marine waters (11 years) and Shark Bay (13 years) were far greater than in the lower Swan River Estuary (6 years). However, the maximum lengths recorded in these three environments were all ca 350 mm. Due to the production by size-related movements of differences amongst the lengths of R. sarba at given ages in different habitats in coastal marine waters, the composite suite of lengths at age was not fully representative of the population of this species as a whole in this environment. A von Bertalanffy growth curve, which was adjusted to take into account size related changes in habitat type, significantly improved the fit to the lengths at age of individuals in the composite samples for the population beyond that provided by the unadjusted von Bertalanffy growth curve. This resulted in the maximum difference between the estimates of length at age from the two growth curves, relative to the L derived from the unadjusted von Bertalanffy curve, reaching a value equivalent to 8 %. However, the maximum differences for the corresponding curves for populations in the lower Swan River Estuary and Shark Bay were far less, i.e. 1.7 and 3.2 %, respectively, and thus not considered biologically significant. Rhabdosargus sarba grew slightly faster in the lower Swan River Estuary than in either coastal marine waters or Shark Bay, possibly reflecting the greater productivity of estuarine environments. Acanthopagrus latus is a protandrous hermaphrodite. Detailed macroscopic and histological examination of the gonads of a wide size range of fish, together with a quantification of how the prevalences of the different categories of gonad change with size and age and during the year, were used to elucidate the sequence of changes that occur in the ovotestes of A. latus during life. The scheme proposed in the present study for the protandrous changes in A. latus differed from those proposed for this species elsewhere, but was similar to that of Pollock (1985) for the congeneric Acanthopagrus australis. The ovotestes of functional males develop from gonads which, as in older juveniles, contain substantial amounts of testicular and ovarian tissue. Such ovotestes, and particularly their testicular component, regress markedly after spawning and then, during the next spawning season, either again become ovotestes in which the testicular zone predominates and contains spermatids and spermatozoa (functional males), or become ovotestes in which the ovarian zone predominates and contains vitellogenic oocytes (functional females). Once a fish has become a functional female, it remains a female throughout the rest of its life. The trends exhibited during the year by reproductive variables demonstrate that A. latus in Shark Bay typically spawns on a very limited number of occasions during a short period in August and September and has determinate fecundity. The potential annual fecundities of 24 A. latus ranged from 764,000 in a 600 g fish to 7,910,000 in a 2,050 g fish and produced a mean [plus-minus]1SE of 1,935,000 [plus-minus] 281,000. The total length at which 50 % of A. latus become identifiable as males (245 mm) is very similar to the current minimum legal length (MLL) of 250 mm, which corresponds to an age of 2.5 years less than the age at which 50 % of males become females. Current spawning potential ratios calculated over a range of alternative values for natural mortality (M) for A. latus in Shark Bay suggests that the present fishing pressure is sustainable, but that the current MLL should be reviewed if recreational fishing pressure continues to increase. The age composition and von Bertalanffy growth parameters for Acanthopagrus latus have been determined. The relevant parameters were inserted into the empirical equations of Pauly (1980) and Ralston (1987) for estimating natural mortality (M). Total mortality (Z) was calculated using Hoenig's (1983) equations, relative abundance analysis and a simulation based on maximum age and sample size.The two point estimates for M for A. latus, which were both 0.70 year-1, greatly exceeded all estimates for Z (range 0.18 to 0.30 year-1), which is clearly an erroneous result. To resolve this problem of inconsistent estimates, a Bayesian approach was developed, which, through combining the likelihood distributions of the various mortality estimates, produced integrated estimates for M and Z that are more consistent and precise than those produced for these two variables using the above methods individually. This approach now yielded lower values for M than Z and a measure of fishing mortality that appears to be consistent with the current status of the fishery. This approach is equally applicable to other fish species.

Various aspects of the biology of the tarwhine Rhabdosargus sarba and western yellowfin bream Acanthopagrus latus were studied. The studies on R. sarba have focused on populations in temperate coastal marine waters at ca 32 degrees S and the lower reaches of an estuary (Swan River Estuary) located at the same latitude and in a subtropical embayment (Shark Bay) at ca 26 degrees S, while those on A. latus were conducted on the population in the latter embayment. A combination of a macroscopic and histological examination of the gonads demonstrated that R. sarba is typically a rudimentary hermaphrodite in Western Australian waters, i.e. the juveniles develop into either a male or female in which the ovarian and testicular zones of the gonads, respectively, are macroscopically undetectable. This contrasts with the situation in the waters off Hong Kong and South Africa, in which R. sarba is reported to be a protandrous hermaphrodite. However, it is possible that a few of the fish that are above the size at first maturity and possess, during the spawning period, ovotestes with relatively substantial amounts of both mature testicular and immature ovarian tissue, could function as males early in adult life and then change to females. Although R. sarba spawns at some time between late winter and late spring in Western Australia, spawning peaks later in the Swan River Estuary than in coastal, marine waters at the same latitude and Shark Bay, in which salinities are always close to or above that of full strength sea water, i.e. 35 0/00. While the males and females attain sexual maturity at very similar lengths in the Swan River Estuary and Shark Bay, i.e. L50s all between 170 and 177 mm, they typically reach maturity at an earlier age in the former environment, i.e. 2 vs 3 years old. Thus, length and consequently growth rate influence the timing of maturity rather than age. During the spawning period, only 9 % of the fish caught between 180 and 260 mm in nearshore, shallow marine waters had become mature, whereas 91 % of those in this length range over reefs were mature, indicating that R. sarba tends to move offshore only when it has become physiologically ready to mature. The L50s at first maturity indicate that the current minimum legal length in Western Australia (230 mm) is appropriate for managing this species. Oocyte diameter frequency distributions, stages in oocyte development, duration of oocyte hydration and time of formation of post-ovulatory follicles in mature ovaries of Rhabdosargus sarba in the lower Swan River Estuary (32 degrees 03'S, 115 degrees 44'E) were used, in conjunction with data on tidal cycles, to elucidate specific aspects of the reproductive biology of this sparid in an estuarine environment. The results demonstrated the following. (i) Rhabdosargus sarba has indeterminate fecundity sensu Hunter et al. (1985). (ii) Oocyte hydration commences at about dusk (18:30 h) and is completed by ca 01:30-04:30 h, at which time ovulation, as revealed by the presence of hydrated oocytes in the ovarian duct and appearance of newlyformed post-ovulatory follicles, commences. (iii) The prevalence of spawning was positively correlated with tidal strength and was greatest on days when the tide changed from flood to ebb at ca 06:00 h, i.e. approximately when spawning ceases. Spawning just prior to strong ebb tides would lead to the transport of eggs out of the estuary and thus into salinities that remain at ca 35 0/00. The likelihood of eggs being transported downstream is further enhanced by R. sarba spawning in deeper waters in the estuary, where the flow is greatest. (iv) Although mature ovaries were found in R. sarba in the estuary between early July and December, the prevalence of atretic oocytes was high until September, when salinities started rising markedly from their winter minima. Batch fecundities ranged from 2,416 for a 188 mm fish to 53,707 for a 266 mm fish. The average daily prevalence of spawning amongst mature females during the spawning period of R. sarba caught in the lower estuary, i.e. July to end of October, was 36.5 %. Thus, individual female R. sarba spawned, on average, at intervals of ca 2.7 days in each spawning season. Female R. sarba with total lengths of 200, 250 and 300 mm were estimated to have a batch fecundity of 7,400, 20,100 and 54,800 eggs, respectively and annual fecundities of 332,000, 903,000 and 2,461,000 eggs, respectively. Rhabdosargus sarba is shown to undergo size-related movements in each of the three very different environments in which it was studied. In temperate coastal waters, R. sarba settles in unvegetated nearshore areas and then moves progressively firstly to nearby seagrass beds and then to exposed unvegetated nearshore areas and finally to areas around reefs where spawning occurs. Although R. sarba spawns in the lower Swan River Estuary, relatively few of its early 0+ recruits remain in the estuary and substantial numbers of this species do not start reappearing in the estuary until they are ca 140 mm. In Shark Bay, R. sarba uses nearshore mangroves as a nursery area and later moves into areas around reefs. The maximum ages recorded for R. sarba in coastal marine waters (11 years) and Shark Bay (13 years) were far greater than in the lower Swan River Estuary (6 years). However, the maximum lengths recorded in these three environments were all ca 350 mm. Due to the production by size-related movements of differences amongst the lengths of R. sarba at given ages in different habitats in coastal marine waters, the composite suite of lengths at age was not fully representative of the population of this species as a whole in this environment. A von Bertalanffy growth curve, which was adjusted to take into account size related changes in habitat type, significantly improved the fit to the lengths at age of individuals in the composite samples for the population beyond that provided by the unadjusted von Bertalanffy growth curve. This resulted in the maximum difference between the estimates of length at age from the two growth curves, relative to the L derived from the unadjusted von Bertalanffy curve, reaching a value equivalent to 8 %. However, the maximum differences for the corresponding curves for populations in the lower Swan River Estuary and Shark Bay were far less, i.e. 1.7 and 3.2 %, respectively, and thus not considered biologically significant. Rhabdosargus sarba grew slightly faster in the lower Swan River Estuary than in either coastal marine waters or Shark Bay, possibly reflecting the greater productivity of estuarine environments. Acanthopagrus latus is a protandrous hermaphrodite. Detailed macroscopic and histological examination of the gonads of a wide size range of fish, together with a quantification of how the prevalences of the different categories of gonad change with size and age and during the year, were used to elucidate the sequence of changes that occur in the ovotestes of A. latus during life. The scheme proposed in the present study for the protandrous changes in A. latus differed from those proposed for this species elsewhere, but was similar to that of Pollock (1985) for the congeneric Acanthopagrus australis. The ovotestes of functional males develop from gonads which, as in older juveniles, contain substantial amounts of testicular and ovarian tissue. Such ovotestes, and particularly their testicular component, regress markedly after spawning and then, during the next spawning season, either again become ovotestes in which the testicular zone predominates and contains spermatids and spermatozoa (functional males), or become ovotestes in which the ovarian zone predominates and contains vitellogenic oocytes (functional females). Once a fish has become a functional female, it remains a female throughout the rest of its life. The trends exhibited during the year by reproductive variables demonstrate that A. latus in Shark Bay typically spawns on a very limited number of occasions during a short period in August and September and has determinate fecundity. The potential annual fecundities of 24 A. latus ranged from 764,000 in a 600 g fish to 7,910,000 in a 2,050 g fish and produced a mean [plus-minus]1SE of 1,935,000 [plus-minus] 281,000. The total length at which 50 % of A. latus become identifiable as males (245 mm) is very similar to the current minimum legal length (MLL) of 250 mm, which corresponds to an age of 2.5 years less than the age at which 50 % of males become females. Current spawning potential ratios calculated over a range of alternative values for natural mortality (M) for A. latus in Shark Bay suggests that the present fishing pressure is sustainable, but that the current MLL should be reviewed if recreational fishing pressure continues to increase. The age composition and von Bertalanffy growth parameters for Acanthopagrus latus have been determined. The relevant parameters were inserted into the empirical equations of Pauly (1980) and Ralston (1987) for estimating natural mortality (M). Total mortality (Z) was calculated using Hoenig's (1983) equations, relative abundance analysis and a simulation based on maximum age and sample size.The two point estimates for M for A. latus, which were both 0.70 year-1, greatly exceeded all estimates for Z (range 0.18 to 0.30 year-1), which is clearly an erroneous result. To resolve this problem of inconsistent estimates, a Bayesian approach was developed, which, through combining the likelihood distributions of the various mortality estimates, produced integrated estimates for M and Z that are more consistent and precise than those produced for these two variables using the above methods individually. This approach now yielded lower values for M than Z and a measure of fishing mortality that appears to be consistent with the current status of the fishery. This approach is equally applicable to other fish species.

Various aspects of the biology of the tarwhine Rhabdosargus sarba and western yellowfin bream Acanthopagrus latus were studied. The studies on R. sarba have focused on populations in temperate coastal marine waters at ca 32oS and the lower reaches of an estuary (Swan River Estuary) located at the same latitude and in a subtropical embayment (Shark Bay) at ca 26oS, while those on A. latus were conducted on the population in the latter embayment. A combination of a macroscopic and histological examination of the gonads demonstrated that R. sarba is typically a rudimentary hermaphrodite in Western Australian waters, i.e. the juveniles develop into either a male or female in which the ovarian and testicular zones of the gonads, respectively, are macroscopically undetectable. This contrasts with the situation in the waters off Hong Kong and South Africa, in which R. sarba is reported to be a protandrous hermaphrodite. However, it is possible that a few of the fish that are above the size at first maturity and possess, during the spawning period, ovotestes with relatively substantial amounts of both mature testicular and immature ovarian tissue, could function as males early in adult life and then change to females. Although R. sarba spawns at some time between late winter and late spring in Western Australia, spawning peaks later in the Swan River Estuary than in coastal, marine waters at the same latitude and Shark Bay, in which salinities are always close to or above that of full strength sea water, i.e. 35 ñ . While the males and females attain sexual maturity at very similar lengths in the Swan River Estuary and Shark Bay, i.e. L50s all between 170 and 177 mm, they typically reach maturity at an earlier age in the former environment, i.e. 2 vs 3 years old. Thus, length and consequently growth rate influence the timing of maturity rather than age. During the spawning period, only 9 % of the fish caught between 180 and 260 mm in nearshore, shallow marine waters had become mature, whereas 91 % of those in this length range over reefs were mature, indicating that R. sarba tends to move offshore only when it has become gphysiologically ready to mature. The L50s at first maturity indicate that the current minimum legal length in Western Australia (230 mm) is appropriate for managing this species. Oocyte diameter frequency distributions, stages in oocyte development, duration of oocyte hydration and time of formation of post-ovulatory follicles in mature ovaries of Rhabdosargus sarba in the lower Swan River Estuary (32o 03fS, 115o 44fE) were used, in conjunction with data on tidal cycles, to elucidate specific aspects of the reproductive biology of this sparid in an estuarine environment. The results demonstrated the following. (i) Rhabdosargus sarba has indeterminate fecundity sensu Hunter et al. (1985). (ii) Oocyte hydration commences at about dusk (18:30 h) and is completed by ca 01:30-04:30 h, at which time ovulation, as revealed by the presence of hydrated oocytes in the ovarian duct and appearance of newlyformed post-ovulatory follicles, commences. (iii) The prevalence of spawning was positively correlated with tidal strength and was greatest on days when the tide changed from flood to ebb at ca 06:00 h, i.e. approximately when spawning ceases. Spawning just prior to strong ebb tides would lead to the transport of eggs out of the estuary and thus into salinities that remain at ca 35 ñ . The likelihood of eggs being transported downstream is further enhanced by R. sarba spawning in deeper waters in the estuary, where the flow is greatest. (iv) Although mature ovaries were found in R. sarba in the estuary between early July and December, the prevalence of atretic oocytes was high until September, when salinities started rising markedly from their winter minima. Batch fecundities ranged from 2,416 for a 188 mm fish to 53,707 for a 266 mm fish. The average daily prevalence of spawning amongst mature females during the spawning period of R. sarba caught in the lower estuary, i.e. July to end of October, was 36.5 %. Thus, individual female R. sarba spawned, on average, at intervals of ca 2.7 days in each spawning season. Female R. sarba with total lengths of 200, 250 and 300 mm were estimated to have a batch fecundity of 7,400, 20,100 and 54,800 eggs, respectively and annual fecundities of 332,000, 903,000 and 2,461,000 eggs, respectively. Rhabdosargus sarba is shown to undergo size-related movements in each of the three very different environments in which it was studied. In temperate coastal waters, R. sarba settles in unvegetated nearshore areas and then moves progressively firstly to nearby seagrass beds and then to exposed unvegetated nearshore areas and finally to areas around reefs where spawning occurs. Although R. sarba spawns in the lower Swan River Estuary, relatively few of its early 0+ recruits remain in the estuary and substantial numbers of this species do not start reappearing in the estuary until they are ca 140 mm. In Shark Bay, R. sarba uses nearshore mangroves as a nursery area and later moves into areas around reefs. The maximum ages recorded for R. sarba in coastal marine waters (11 years) and Shark Bay (13 years) were far greater than in the lower Swan River Estuary (6 years). However, the maximum lengths recorded in these three environments were all ca 350 mm. Due to the production by size-related movements of differences amongst the lengths of R. sarba at given ages in different habitats in coastal marine waters, the composite suite of lengths at age was not fully representative of the population of this species as a whole in this environment. A von Bertalanffy growth curve, which was adjusted to take into account size related changes in habitat type, significantly improved the fit to the lengths at age of individuals in the composite samples for the population beyond that provided by the unadjusted von Bertalanffy growth curve. This resulted in the maximum difference between the estimates of length at age from the two growth curves, relative to the L‡ derived from the unadjusted von Bertalanffy curve, reaching a value equivalent to 8 %. However, the maximum differences for the corresponding curves for populations in the lower Swan River Estuary and Shark Bay were far less, i.e. 1.7 and 3.2 %, respectively, and thus not considered biologically significant. Rhabdosargus sarba grew slightly faster in the lower Swan River Estuary than in either coastal marine waters or Shark Bay, possibly reflecting the greater productivity of estuarine environments. Acanthopagrus latus is a protandrous hermaphrodite. Detailed macroscopic and histological examination of the gonads of a wide size range of fish, together with a quantification of how the prevalences of the different categories of gonad change with size and age and during the year, were used to elucidate the sequence of changes that occur in the ovotestes of A. latus during life. The scheme proposed in the present study for the protandrous changes in A. latus differed from those proposed for this species elsewhere, but was similar to that of Pollock (1985) for the congeneric Acanthopagrus australis. The ovotestes of functional males develop from gonads which, as in older juveniles, contain substantial amounts of testicular and ovarian tissue. Such ovotestes, and particularly their testicular component, regress markedly after spawning and then, during the next spawning season, either again become ovotestes in which the testicular zone predominates and contains spermatids and spermatozoa (functional males), or become ovotestes in which the ovarian zone predominates and contains vitellogenic oocytes (functional females). Once a fish has become a functional female, it remains a female throughout the rest of its life. The trends exhibited during the year by reproductive variables demonstrate that A. latus in Shark Bay typically spawns on a very limited number of occasions during a short period in August and September and has determinate fecundity. The potential annual fecundities of 24 A. latus ranged from 764,000 in a 600 g fish to 7,910,000 in a 2,050 g fish and produced a mean }1SE of 1,935,000 } 281,000. The total length at which 50 % of A. latus become identifiable as males (245 mm) is very similar to the current minimum legal length (MLL) of 250 mm, which corresponds to an age of 2.5 years less than the age at which 50 % of males become females. Current spawning potential ratios calculated over a range of alternative values for natural mortality (M) for A. latus in Shark Bay suggests that the present fishing pressure is sustainable, but that the current MLL should be reviewed if recreational fishing pressure continues to increase. The age composition and von Bertalanffy growth parameters for Acanthopagrus latus have been determined. The relevant parameters were inserted into the empirical equations of Pauly (1980) and Ralston (1987) for estimating natural mortality (M). Total mortality (Z) was calculated using Hoenigfs (1983) equations, relative abundance analysis and a simulation based on maximum age and sample size.The two point estimates for M for A. latus, which were both 0.70 year-1, greatly exceeded all estimates for Z (range 0.18 to 0.30 year-1), which is clearly an erroneous result. To resolve this problem of inconsistent estimates, a Bayesian approach was developed, which, through combining the likelihood distributions of the various mortality estimates, produced integrated estimates for M and Z that are more consistent and precise than those produced for these two variables using the above methods individually. This approach now yielded lower values for M than Z and a measure of fishing mortality that appears to be consistent with the current status of the fishery. This approach is equally applicable to other fish species.

碩士
國立臺北科技大學
土木與防災研究所
101
This study analyzed through numerical method to simulate materials in triaxial (including single pressure) mechanical behavior under test conditions, and for the microscopic parameters for changing the material circumstances, the corresponding macroscopic parameters as discussed 　　This study is based on the Distinct Element Method Numerical software PFC3D Simulation microscopic spherical particles between elements of the mechanical behavior of non-continuum, and to establish the elements of microscopic parameters and the overall macroscopic material parameter sensitivity analysis of interconnectedness. In triaxial tests, for example, elements through microscopic particles parameter settings for PFC3D the simulation results obtained under different microscopic parameters set corresponding overall macroscopic parameters, in respect material of micro and macro parameter checking investigate the association , That material macroscopic elastic modulus and uniaxial compressive strength, respectively, with the microscopic particle elements elastic modulus and bond strength between particles is proportional approximately,another material macroscopic parameters - Poisson''s ratio is positively related to particles of the elements of positive / tangential stiffness ratio. Used statistical analysis software-SPSS as analysis using regression methods, represent between the micro, macro parameter relationships by regression equation, in future PFC3D simulated particles and bonding material parameters such as the alignment of various microscopic and macroscopic materials to meet the desired simulated parameters reference direction of research work.

Leung Ling Yan.
Thesis (Ph.D.)--Chinese University of Hong Kong, 2009.
Includes bibliographical references (leaves 218-259).
Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
Abstract also in Chinese.

Man, Ka Yan.
Thesis (M.Phil.)--Chinese University of Hong Kong, 2008.
Includes bibliographical references (leaves 101-126).
Abstracts in English and Chinese.
Chapter I. --- Title page --- p.i
Chapter II. --- Thesis committee --- p.ii
Chapter III. --- Acknowledgements --- p.iii
Chapter IV. --- Abstract (English) --- p.iv
Chapter V. --- Abstract (Chinese) --- p.vi
Chapter VI. --- Table of contents --- p.vii
Chapter VII. --- List of abbreviations --- p.xiii
Chapter VIII. --- List of figures --- p.xv
General introduction --- p.1
Chapter Chapter 1: --- Literature review --- p.4
Chapter 1.1. --- Cadmium --- p.5
Chapter 1.1.1. --- Cadmium - Ways of uptake in human and aquatic life --- p.5
Chapter 1.1.2. --- Cadmium - Toxic effects in fish --- p.6
Chapter 1.2. --- Cortisol --- p.11
Chapter 1.2.1. --- Cortisol - General information and its regulations --- p.11
Chapter 1.2.2. --- Cortisol - Functions --- p.12
Chapter 1.3. --- Thyroid hormones --- p.14
Chapter 1.3.1. --- THs - General information and its regulations --- p.14
Chapter 1.3.2. --- THs - Functions --- p.15
Chapter 1.4. --- Growth hormone --- p.18
Chapter 1.4.1. --- GH - General information and its regulations --- p.18
Chapter 1.4.2. --- GH - Functions --- p.20
Chapter 1.5. --- Insulin-like growth factor --- p.22
Chapter 1.5.1. --- IGF-I - General information and its regulations --- p.22
Chapter 1.5.2. --- IGF-I - Functions --- p.24
Chapter 1.6 --- Metallothioneins --- p.26
Chapter 1.6.1. --- MTs - Definition and Classification --- p.26
Chapter 1.6.2. --- MTs - Functions --- p.27
Chapter 1.7. --- Glucose-6-phosphate dehydrogenase --- p.31
Chapter 1.7.1. --- G6PDH - General information and its regulations --- p.31
Chapter 1.7.2. --- G6PDH ´ؤ Functions --- p.32
Chapter Chapter 2: --- "Effects of cadmium exposure on the endocrine status of silver sea bream, Sparus sarba" --- p.34
Chapter 2.1. --- Introduction --- p.35
Chapter 2.2. --- Materials and methods --- p.37
Chapter 2.2.1. --- Overall experimental design --- p.37
Chapter 2.2.2. --- In vivo exposure to waterborne cadmium --- p.37
Chapter 2.2.2.1. --- Experimental animals --- p.37
Chapter 2.2.2.2. --- Adaptation --- p.37
Chapter 2.2.2.3. --- Tissue sampling --- p.38
Chapter 2.2.2.4. --- Enzyme-linked immunosorbent assay (ELISA) --- p.38
Chapter 2.2.2.4.1. --- Serum cortisol analysis --- p.39
Chapter 2.2.2.4.2. --- Serum triiodothyronine analysis --- p.39
Chapter 2.2.2.4.3. --- Serum thyroxine analysis --- p.39
Chapter 2.2.2.5. --- Protein extraction and Protein quantification --- p.40
Chapter 2.2.2.6. --- Protein gel electrophoresis and immunoblotting (Western blotting) --- p.40
Chapter 2.2.2.7. --- RNA extraction --- p.41
Chapter 2.2.2.8. --- Reverse transcription for first-strand cDNAs from total RNAs samples from liver --- p.42
Chapter 2.2.2.9. --- Real-time quantitative PCR assays of IGF-I mRNA expression --- p.43
Chapter 2.2.3. --- In vivo experiments involving cadmium injection --- p.44
Chapter 2.2.3.1. --- Experimental animals --- p.44
Chapter 2.2.3.2. --- Adaptation --- p.44
Chapter 2.2.3.3. --- Tissue sampling --- p.45
Chapter 2.2.3.4. --- Enzyme-linked immunosorbent assay (ELISA) --- p.45
Chapter 2.2.3.4.1. --- Serum cortisol analysis --- p.45
Chapter 2.2.3.4.2. --- Serum triiodothyronine analysis --- p.45
Chapter 2.2.3.4.3. --- Serum thyroxine analysis --- p.46
Chapter 2.2.3.5. --- Protein extraction and Protein quantification --- p.46
Chapter 2.2.3.6. --- Protein gel electrophoresis and immunoblotting (Western blotting) --- p.46
Chapter 2.2.3.7. --- RNA extraction --- p.46
Chapter 2.2.3.8. --- Reverse transcription for the first-strand cDNAs from total RNAs samples from liver --- p.46
Chapter 2.2.3.9. --- Real-time quantitative PCR of IGF-I mRNA expression --- p.46
Chapter 2.2.4. --- In vitro part of the project (Primary cell culture: hepatocytes exposed to cadmium medium) --- p.47
Chapter 2.2.4.1. --- Experimental animals --- p.47
Chapter 2.2.4.2. --- Primary hepatocytes culture preparation --- p.47
Chapter 2.2.4.3. --- Cadmium treatment and cell harvest --- p.48
Chapter 2.2.4.4. --- "RNA extraction, reverse transcription for the first-strand cDNAs from total RNAs samples from lysed cells and real-time quantitative PCR of IGF-I mRNA expression" --- p.48
Chapter 2.2.5. --- Statistical analysis --- p.48
Chapter 2.3. --- Results --- p.49
Chapter 2.3.1. --- Serum cortisol level --- p.49
Chapter 2.3.2. --- Serum triiodothyronine level --- p.49
Chapter 2.3.3. --- Serum thyroxine level --- p.49
Chapter 2.3.4. --- Pituitary growth hormone level --- p.50
Chapter 2.3.5. --- Hepatic insulin-like growth factor mRNA expression --- p.50
Chapter 2.4. --- Discussion --- p.58
Chapter 2.4.1. --- Serum cortisol level --- p.58
Chapter 2.4.2. --- Thyroid hormones --- p.61
Chapter 2.4.3. --- Growth hormone --- p.64
Chapter 2.4.4. --- Insulin-like growth factor-I --- p.67
Chapter 2.5. --- Conclusion --- p.70
Chapter Chapter 3: --- "Effects of cadmium exposure on MT and G6PDH mRNA expressions of silver sea bream, Sparus sarba" --- p.71
Chapter 3.1. --- Introduction --- p.72
Chapter 3.2. --- Materials and methods --- p.74
Chapter 3.2.1. --- Overall experimental design --- p.74
Chapter 3.2.2. --- In vivo experiments involving exposure to waterborne cadmium --- p.74
Chapter 3.2.2.1. --- Experimental animals --- p.74
Chapter 3.2.2.2. --- Adaptation --- p.74
Chapter 3.2.2.3. --- Tissue sampling --- p.74
Chapter 3.2.2.4. --- RNA extraction --- p.74
Chapter 3.2.2.5. --- Reverse transcription for first-strand cDNAs from total RNAs samples from gill and liver --- p.75
Chapter 3.2.2.6. --- Amplification of partial fragments of metallothionein (MT) --- p.75
Chapter 3.2.2.7. --- Rapid amplification of 5´ة and 3´ة cDNA ends of metallothionein (MT) --- p.76
Chapter 3.2.2.7.1. --- Amplification of 5´ة cDNA end --- p.76
Chapter 3.2.2.7.2. --- Amplification of 3´ة cDNA end --- p.78
Chapter 3.2.2.8. --- Real-time quantitative PCR of MT and G6PDH mRNA expressions --- p.79
Chapter 3.2.3. --- In vivo injection of cadmium --- p.80
Chapter 3.2.3.1. --- "Experimental animals, adaptation and tissue sampling" --- p.80
Chapter 3.2.3.2. --- RNA extraction --- p.80
Chapter 3.2.3.3. --- Reverse transcription for first-strand cDNAs from total RNAs samples from gill and liver --- p.81
Chapter 3.2.3.4. --- Real-time quantitative PCR of MT and G6PDH mRNA expression --- p.81
Chapter 3.2.4. --- In vitro exposure of primary hepatocyte culture to cadmium --- p.81
Chapter 3.2.4.1. --- Experimental animals --- p.81
Chapter 3.2.4.2. --- Preparation of the hepatocytes for cell culture --- p.81
Chapter 3.2.4.3. --- Cadmium treatment and cell harvest --- p.81
Chapter 3.2.4.4. --- "RNA extraction, reverse transcription for first-strand cDNAs from total RNAs samples from lysed cells and real-time quantitative PCR of MT and G6PDH mRNA expression" --- p.82
Chapter 3.2.5. --- Statistical analysis --- p.82
Chapter 3.3. --- Results --- p.83
Chapter 3.3.1. --- MT cloning and characterization --- p.83
Chapter 3.3.2. --- Metallothionein mRNA expression --- p.83
Chapter 3.3.3. --- Hepatic glucose-6-phosphate dehydrogenase mRNA expression --- p.84
Chapter 3.4. --- Discussion --- p.91
Chapter 3.4.1. --- MT cloning and characterization --- p.91
Chapter 3.4.2. --- Metallothioneins mRNA expression --- p.92
Chapter 3.4.3. --- Hepatic glucose-6-phosphate dehydrogenase mRNA expression --- p.95
Chapter 3.5. --- Conclusion --- p.98
General conclusion --- p.99
References --- p.101

Yuen, Wing Sum.
Thesis (M.Phil.)--Chinese University of Hong Kong, 2009.
Includes bibliographical references (leaves 136-155).
Abstract also in Chinese.
Chapter I --- Title page --- p.i
Chapter II --- Acknowledgements --- p.ii
Chapter III --- Abstract --- p.iii
Chapter IV --- Abstract (Chinese version) --- p.vi
Chapter V --- Table of contents --- p.viii
Chapter VI --- List of abbreviations --- p.xv
Chapter VII --- List of figures --- p.xvi
Chapter Chapter 1 --- General introduction --- p.1
Chapter Chapter 2 --- Literature review --- p.5
Chapter 2.1 --- Cystic fibrosis transmembrane conductance regulator in human --- p.5
Chapter 2.1.1. --- Pathology of cystic fibrosis --- p.5
Chapter 2.1.2. --- CFTR gene and the encoded protein --- p.6
Chapter 2.1.3. --- Hypothetical model for CFTR function --- p.7
Chapter 2.1.4. --- Functions of CFTR --- p.7
Chapter 2.1.5. --- Regulation of CFTR gene expression --- p.8
Chapter 2.1.6 --- Regulation of CFTR protein --- p.9
Chapter 2.1.7. --- Discovery of CFTR homologues in other vertebrates --- p.10
Chapter 2.2 --- Cystic fibrosis transmembrane conductance regulator in teleosts --- p.10
Chapter 2.2.1. --- Evidence for the presence of CFTR homologue in teleosts --- p.10
Chapter 2.2.2. --- Molecular cloning of teleost CFTR genes --- p.11
Chapter 2.2.3. --- Role of teleost CFTR in osmoregulation --- p.13
Chapter 2.2.3.1. --- Tissue distribution of CFTR --- p.13
Chapter 2.2.3.2. --- Changes in CFTR expression in response to ambient salinities --- p.14
Chapter 2.2.3.3. --- Immunocytochemical studies of CFTR --- p.15
Chapter 2.2.3.4. --- Regulation of CFTR --- p.17
Chapter 2.3 --- Osmoregulation in teleosts --- p.19
Chapter 2.3.1. --- Importance of osmoregulation --- p.19
Chapter 2.3.2. --- Major components of chloride cells in marine teleosts --- p.20
Chapter 2.3.2.1. --- Overview --- p.20
Chapter 2.3.2.2. --- Sodium-potassium adenosine triphosphatase (Na+，K+-ATPase) --- p.21
Chapter 2.3.2.3. --- Cystic fibrosis transmembrane conductance regulator (CFTR) --- p.22
Chapter 2.3.2.4. --- Na+/K+/2Cr cotransporter (NKCC) --- p.23
Chapter 2.3.2.5. --- Potassium (K+) channel --- p.25
Chapter 2.4 --- Endocrine control of osmoregulation --- p.26
Chapter 2.4.1. --- Overview --- p.26
Chapter 2.4.2. --- Growth hormone (GH) and insulin-like growth factor I (IGF-I) --- p.27
Chapter 2.4.2.1. --- Role of GH in osmoregulation --- p.27
Chapter 2.4.2.2. --- Mediation through IGF-I --- p.29
Chapter 2.4.2.3. --- Synergic effect with cortisol --- p.30
Chapter 2.4.3. --- Prolactin (PRL) --- p.30
Chapter 2.4.3.1. --- Role of PRL in osmoregulation --- p.30
Chapter 2.4.3.2. --- Synergic effect with cortisol --- p.33
Chapter 2.4.4. --- Cortisol --- p.33
Chapter 2.4.4.1. --- Role of cortisol in osmoregulation --- p.33
Chapter 2.4.4.2. --- Dual functions of cortisol --- p.34
Chapter Chapter 3 --- Cloning and tissue distribution of silver sea bream CFTR gene --- p.36
Chapter 3.1 --- Introduction --- p.36
Chapter 3.2 --- Materials and methods --- p.38
Chapter 3.2.1. --- Part A: Cloning of silver sea bream CFTR gene --- p.38
Chapter 3.2.1.1. --- Fish and culture conditions --- p.38
Chapter 3.2.1.2. --- Sampling of fish --- p.38
Chapter 3.2.1.3. --- Preparation of first strand cDNA --- p.38
Chapter 3.2.1.4. --- Design of primers --- p.39
Chapter 3.2.1.5. --- Semi-quantitative reverse transcriptase (RT)-PCR --- p.40
Chapter 3.2.1.6 --- Cloning --- p.41
Chapter 3.2.2. --- Part B: Tissue distribution of CFTR in silver sea bream --- p.41
Chapter 3.2.2.1. --- Fish and culture conditions --- p.41
Chapter 3.2.2.2. --- Tissue sampling --- p.42
Chapter 3.2.2.3. --- Preparation of first strand cDNA --- p.42
Chapter 3.2.2.4 --- Design of primers --- p.42
Chapter 3.2.2.5. --- Semi-quantitative reverse transcriptase (RT)-PCR --- p.43
Chapter 3.3 --- Results --- p.44
Chapter 3.3.1. --- Part A: Cloning of silver sea bream CFTR gene --- p.44
Chapter 3.3.2. --- Part B: Tissue distribution of CFTR in silver sea bream --- p.60
Chapter 3.4 --- Discussion --- p.62
Chapter 3.4.1. --- Part A: Cloning of silver sea bream CFTR --- p.62
Chapter 3.4.2. --- Part B: Tissue distribution of CFTR in silver sea bream --- p.64
Chapter Chapter 4 --- Effect of salinity on CFTR mRNA expression in gill and posterior intestine of silver sea bream --- p.68
Chapter 4.1 --- Introduction --- p.68
Chapter 4.2 --- Materials and methods --- p.70
Chapter 4.2.1. --- Part A: Effect of long-term exposure to different salinities on CFTR expression --- p.70
Chapter 4.2.1.1. --- Experimental fish and salinity adaptation --- p.70
Chapter 4.2.1.2. --- Tissue sampling --- p.70
Chapter 4.2.1.3. --- Serum ion levels --- p.71
Chapter 4.2.1.4. --- Preparation of first strand cDNA --- p.71
Chapter 4.2.1.5. --- Design of primers --- p.71
Chapter 4.2.1.6. --- Semi-quantitative reverse transcriptase (RT)-PCR --- p.71
Chapter 4.2.1.7. --- Statistical analysis --- p.72
Chapter 4.2.2. --- Part B: Effect of abrupt transfer on CFTR expression --- p.73
Chapter 4.2.2.1. --- Experimental fish --- p.73
Chapter 4.2.2.2. --- Experimental design --- p.73
Chapter 4.2.2.2.1 --- Experiment 1: Abrupt transfer from seawater (SW) to 6 ppt --- p.73
Chapter 4.2.2.2.2. --- Experiment 2: Abrupt transfer from 6 ppt to SW --- p.73
Chapter 4.2.2.3. --- Tissue sampling --- p.74
Chapter 4.2.2.4. --- Serum ion levels --- p.74
Chapter 4.2.2.5. --- Preparation of first strand cDNA --- p.74
Chapter 4.2.2.6. --- Design of primers --- p.75
Chapter 4.2.2.7. --- Semi-quantitative reverse transcriptase (RT)-PCR --- p.75
Chapter 4.2.2.8. --- Statistical analysis --- p.75
Chapter 4.3 --- Results --- p.76
Chapter 4.3.1. --- Part A: Effect of long-term exposure to different salinities on CFTR expression --- p.76
Chapter 4.3.1.1. --- Serum ion levels --- p.76
Chapter 4.3.1.2. --- CFTR expression in gill --- p.76
Chapter 4.3.1.3. --- CFTR expression in posterior intestine --- p.76
Chapter 4.3.2. --- Part B: Effect of abrupt salinity transfer on CFTR expression --- p.83
Chapter 4.3.2.1. --- Experiment 1: Abrupt transfer from SW to 6 ppt --- p.83
Chapter 4.3.2.1.1. --- Serum ion levels --- p.83
Chapter 4.3.2.1.2. --- CFTR in gill --- p.83
Chapter 4.3.2.1.3. --- CFTR in posterior intestine --- p.83
Chapter 4.3.2.2. --- Experiment 2: Abrupt transfer from 6 ppt to SW --- p.89
Chapter 4.3.2.2.1. --- Serum ion levels --- p.89
Chapter 4.3.2.2.2. --- CFTR in gill --- p.89
Chapter 4.3.2.2.3. --- CFTR in posterior intestine --- p.89
Chapter 4.4 --- Discussion --- p.95
Chapter 4.4.1. --- Long-term exposure to various salinities --- p.95
Chapter 4.4.2. --- Abrupt salinity transfer --- p.98
Chapter 4.4.2.1. --- Abrupt hypo-osmotic transfer (33 ppt to 6 ppt) --- p.98
Chapter 4.4.2.2. --- Abrupt seawater transfer (6 ppt to 33 ppt) --- p.99
Chapter 4.4.3. --- CFTR mRNA expression in posterior intestine --- p.101
Chapter 4.4.4. --- Conclusion --- p.101
Chapter Chapter 5 --- Effect of hormones on CFTR expression in gill and posterior intestine of silver sea bream --- p.102
Chapter 5.1 --- Introduction --- p.102
Chapter 5.2 --- Materials and methods --- p.104
Chapter 5.2.1. --- Part A: In vivo effect of hormones on CFTR expression --- p.104
Chapter 5.2.1.1. --- Experimental fish and salinity adaptation --- p.104
Chapter 5.2.1.2. --- Hormone treatment --- p.104
Chapter 5.2.1.3. --- Tissue sampling --- p.105
Chapter 5.2.1.4. --- "Serum ion levels, preparation of first strand cDNA, design of primers and semi-quantitative reverse transcriptase (RT)-PCR" --- p.105
Chapter 5.2.1.5. --- Statistical analysis --- p.105
Chapter 5.2.2. --- Part B: In vitro effect of hormones on CFTR expression --- p.106
Chapter 5.2.2.1. --- Fish and culture conditions --- p.106
Chapter 5.2.2.2. --- Gill and posterior intestine preparations --- p.106
Chapter 5.2.2.3. --- Hormone treatment --- p.106
Chapter 5.2.2.4. --- "Preparation of first strand cDNA, design of primers and semi-quantitative reverse transcriptase (RT)-PCR" --- p.107
Chapter 5.2.2.5. --- Statistical analysis --- p.107
Chapter 5.3 --- Results --- p.108
Chapter 5.3.1. --- Part A: In vivo effect of hormones on CFTR expression --- p.108
Chapter 5.3.1.1. --- Serum ion levels --- p.108
Chapter 5.3.1.1.1. --- Serum [Na+] level --- p.108
Chapter 5.3.1.1.2. --- Serum [K+] level --- p.108
Chapter 5.3.1.1.3. --- Serum [Cl' ] level --- p.108
Chapter 5.3.1.2. --- CFTR expression in gill --- p.109
Chapter 5.3.1.3. --- CFTR expression in posterior intestine --- p.109
Chapter 5.3.2. --- Part B: In vitro effect of hormones on CFTR expression --- p.115
Chapter 5.3.2.1. --- CFTR expression in gill --- p.115
Chapter 5.3.2.2. --- CFTR expression in posterior intestine --- p.115
Chapter 5.4 --- Discussion --- p.122
Chapter 5.4.1. --- Effects of cortisol on CFTR expression --- p.122
Chapter 5.4.2. --- Effects of growth hormone on CFTR expression --- p.124
Chapter 5.4.3. --- Effects of prolactin on CFTR expression --- p.127
Chapter 5.4.4. --- "Overall effect of cortisol, growth hormone and prolactin on CFTR expression" --- p.128
Chapter 5.4.5 --- Conclusion --- p.130
Chapter Chapter 6 --- General discussion and conclusion --- p.132
References --- p.136

Branchial carbonic anhydrase was purified from silver seabream (Sparus sarba) and antibody against the enzyme was obtained by immunization in rabbits. An assay for quantifying the activity of carbonic anhydrase was developed. Using enzymatic and immunological techniques, the activity, expression and distribution of branchial carbonic anhydrase of silver seabream acclimated to different salinities were studied. Fish gill is one of the most important organs involved in various homeostatic processes. The ability of euryhaline fish to maintain constant internal ionic balance is crucial for the survival of the fish upon change in salinity. The presence of carbonic anhydrase in the chloride cells was suggested to be an important enzyme involved in ion regulation of fish.
In the present study, branchial carbonic anhydrase and erythrocyte carbonic anhydrase were purified from the gill cells of silver seabream with p-aminomethylbenzenesulfonamide-agarose affinity column. They were predominantly cytosolic with a molecular size of 26.6 k Da for branchial carbonic anhydrase and 28.6 k Da for erythrocyte carbonic anhydrase. Investigation of kinetic properties towards the inhibitor acetazolamide has helped determine the inhibition constants (Ki of branchial carbonic anhydrase: 0.54 x 10-9; Ki of erythrocyte carbonic anhydrase: 0.22 x 10-9). The difference in molecular size and inhibition constant towards acetazolamide supported the view that branchial carbonic anhydrase and erythrocyte carbonic anhydrase were two different isozymes. Polyclonal antibody specific to seabream branchial carbonic anhydrase was obtained by immunization in rabbit. The distribution of branchial carbonic anhydrase in the gill of seabream acclimated to different salinities was studied with immunohistochemical method. The enzyme was mainly located at the interlamellar region. The effect of salinity (0, 6, 12, 33, 50 and 70 ‰) acclimation on the expression and activities of branchial carbonic anhydrase has shown a U-shape pattern from freshwater to double-strength seawater on the quantity of seabream branchial carbonic anhydrase. Higher amount of branchial carbonic anhydrase in freshwater was consistent with the current view that the enzyme was actively involved in the ion uptake process through the hydration of carbon dioxide to produce bicarbonate ion and proton for the exchange of chloride and sodium ions, respectively. An interesting finding was obtained with elevated amount of branchial carbonic anhydrase in seabream acclimated to double-strength seawater and the possible role of the enzyme in such extreme environment was discussed.
This study has provided useful information on the properties, localizations and activities of branchial carbonic anhydrase in silver seabream for the understanding of the involvement of the enzyme in salinity adaptation of silver seabream.
Ma, Wing Chi Joyce.
Source: Dissertation Abstracts International, Volume: 70-06, Section: B, page: 3250.
Thesis (Ph.D.)--Chinese University of Hong Kong, 2008.
Includes bibliographical references (leaves 127-151).
Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
Abstracts in English and Chinese.
School code: 1307.

by Scott P. Kelly.
Thesis (Ph.D.)--Chinese University of Hong Kong, 1998.
Includes bibliographical references (p. 378-410).
Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
Mode of access: World Wide Web.

碩士
國立中山大學
企業管理學系研究所
95
Under the development trend of globalization, enterprises are forced to follow the vision of corporate governance. Enterprises must construct an effective internal controlling system in the basic running environment, and strengthen the enterprises risk management, follow the SOX norm to improve the corporate image then march toward the internationalization. This research adopts the case study, choosing the representative enterprise, the ASE Incorporated. By the way of depth interview and analysis in the relational papers, we have the main conclusions as the result as follows. First, Strength the stratagems of corporate governance: Raising the information transparency, establishing the internal management process and strengthen the enterprises risk management, etc. The ASE Incorporated combines the SOX and internal control norm, in order to make the result of resources integration. Second, balance the legal systems and practices of independent director: Shown in the secondary materials and analyze other information of this research, the legal systems and practices are not in opposing side. The considerations of setting up the independent director can improve the quality of decision and raise the effects, but do not influence the decision-making by way of full powers. However, it can intensify the governance efficiency. We make some suggestions for the enterprises. It is including place importance of internal control system and internal audit management and enhancement the information system. The good institutional framework can be expected the development goal of corporate governance.

Zhang, Chaoxiong.
Thesis (M.Phil.)--Chinese University of Hong Kong, 2010.
Includes bibliographical references (leaves 115-132).
Abstracts in English and Chinese.
Chapter I --- Title page --- p.i
Chapter II --- Thesis committee --- p.ii
Chapter III --- Abstract --- p.iii
Chapter IV --- Abstract (Chinese version) --- p.v
Chapter V --- Acknowledgement --- p.vii
Chapter VI --- Table of content --- p.viii
Chapter VII --- List of figure --- p.xiii
Chapter Chapter 1 --- General introduction --- p.1
Chapter Chapter 2 --- Literature review --- p.7
Chapter 2.1 --- An introduction to myostatin --- p.8
Chapter 2.1.1 --- A general introduction --- p.8
Chapter 2.1.2 --- Myostatin identification --- p.9
Chapter 2.1.3 --- Structural studies of myostatin --- p.10
Chapter 2.1.4 --- Phenotype of myostatin-null animals or transgenic animal --- p.10
Chapter 2.2 --- Regulation of myostatin --- p.12
Chapter 2.2.1 --- Biosynthesis of myostatin --- p.12
Chapter 2.2.2 --- Regulation of myostatin expression --- p.13
Chapter 2.2.3 --- Regulation of myostatin protein --- p.16
Chapter 2.3 --- Myostatin effect --- p.20
Chapter 2.3.1 --- Myostatin Signaling Pathway --- p.20
Chapter 2.3.2 --- Cellular Responses to Myostatin Signaling --- p.23
Chapter 2.4 --- Possible functions in tissues other than muscle --- p.26
Chapter 2.5 --- Myostatin in fishes --- p.27
Chapter 2.5.1 --- Introduction of silver sea bream --- p.27
Chapter 2.5.2 --- Studies carried out in fishes --- p.27
Chapter 2.5.3 --- Possible novel functions of myostatin in fishes --- p.30
Chapter Chapter 3 --- Characterization of myostatin gene in the silver seabream (Sparus sarba) --- p.31
Chapter 3.1 --- Abstract --- p.32
Chapter 3.2 --- Introduction --- p.33
Chapter 3.3 --- Materials and methods --- p.35
Chapter 3.3.1 --- Experimental fish --- p.35
Chapter 3.3.2 --- Total RNA extraction and cDNA cloning of myostatin-1 and myostatin-2 in silver sea bream --- p.35
Chapter 3.3.3 --- Multiple sequence alignment --- p.38
Chapter 3.3.4 --- Real-time PCR for quantification of myostatin-1 and myostatin-2 mRNA expression --- p.38
Chapter 3.3.5 --- 1 --- p.39
Chapter 3.3.6 --- Data processing and statistical analysis --- p.40
Chapter 3.4 --- Results --- p.40
Chapter 3.4.1 --- Cloning of myostatin-l and myostatin-2 cDNA --- p.40
Chapter 3.4.2 --- Myostatin tissue distribution and seasonal pattern --- p.42
Chapter 3.5 --- Discussion --- p.55
Chapter Chapter 4 --- "Effects of growth hormone, 11-ketotestosterone and cortisol on myostatin mRNA expression in silver sea bream (Sparus sarba)" --- p.61
Chapter 4.1 --- Abstract --- p.62
Chapter 4.2 --- Introduction --- p.63
Chapter 4.3 --- Materials and methods --- p.65
Chapter 4.3.1 --- Experimental fish --- p.65
Chapter 4.3.2 --- Growth hormone injection --- p.65
Chapter 4.3.3 --- 11-ketotestosterone and cortisol injection --- p.66
Chapter 4.3.4 --- Muscle explants culture and hormone exposure --- p.67
Chapter 4.3.5 --- Primary pituitary cell culture and cortisol exposure --- p.68
Chapter 4.3.6 --- Measurement of growth hormone secretion by ELISA --- p.69
Chapter 4.3.7 --- Data processing and statistical analysis --- p.70
Chapter 4.4 --- Results --- p.71
Chapter 4.4.1 --- Growth hormone injection --- p.71
Chapter 4.4.2 --- 11-ketotestosterone injection --- p.71
Chapter 4.4.3 --- Cortisol injection --- p.71
Chapter 4.4.4 --- "In vitro hormone treatment-growth hormone, 11-ketotestosterone and cortisol" --- p.72
Chapter 4.4.5 --- Pituitary cell growth hormone secretion under cortisol treatment --- p.72
Chapter 4.5 --- Discussion --- p.81
Chapter Chapter 5 --- Expression of myostatin mRNA in silver sea bream in different salinity --- p.87
Chapter 5.1 --- Abstract --- p.88
Chapter 5.2 --- Introduction --- p.89
Chapter 5.3 --- Materials and Methods --- p.91
Chapter 5.3.1 --- Experimental fish --- p.92
Chapter 5.3.2 --- Long term salinity adaptation --- p.92
Chapter 5.3.3 --- Abrupt transfer form seawater to freshwater --- p.92
Chapter 5.3.4 --- Data processing and statistical analysis --- p.93
Chapter 5.4 --- Results --- p.93
Chapter 5.4.1 --- Long term adaptation to different salinities --- p.93
Chapter 5.4.2 --- Abrupt transfer from 33ppt to 6ppt - 24 h --- p.93
Chapter 5.4.3 --- Abrupt transfer from 33ppt to 6ppt - 72 h --- p.94
Chapter 5.5 --- Discussion --- p.104
Chapter Chapter 6 --- General discussion and conclusion --- p.108
References --- p.115

Chau, Kai Ming.
Thesis (M.Phil.)--Chinese University of Hong Kong, 2009.
Includes bibliographical references (leaves 136-159).
Abstract also in Chinese.
Chapter I --- Abstract --- p.i
Chapter II --- Acknowledgements --- p.vi
Chapter III --- Table of Contents --- p.vii
Chapter IV --- List of Figures --- p.xv
Chapter Chapter 1: --- Introduction --- p.1
Chapter Chapter 2: --- Literature review --- p.7
Chapter 2.1 --- Na+-K+ ATPase --- p.7
Chapter 2.1.1 --- Introduction
Chapter 2.1.2 --- Structure of Na+-K+ ATPase --- p.9
Chapter 2.1.1.2 --- Na+-K+ ATPase a subunit --- p.9
Chapter 2.1.1.3 --- Na+-K+ ATPase β subunit --- p.11
Chapter 2.1.1.4 --- Composition of the a subunit and β subunit --- p.12
Chapter 2.1.1.5 --- Isomers of Na+-K+ ATPase --- p.13
Chapter 2.1.1.6 --- Mechanism of ion exchange --- p.15
Chapter 2.2 --- Aquaporins --- p.17
Chapter 2.2.1 --- Introduction --- p.17
Chapter 2.2.2 --- Structure of AQP-1 --- p.18
Chapter 2.2.3 --- Distribution and function of AQP-1 --- p.19
Chapter 2.3 --- Hormone --- p.22
Chapter 2.3.1 --- Prolactin --- p.22
Chapter 2.3.1.1 --- Structure of prolactin --- p.22
Chapter 2.3.1.2. --- Functions of prolactin --- p.24
Chapter 2.3.2 --- Growth hormone --- p.27
Chapter 2.3.2.1 --- Structure --- p.27
Chapter 2.3.2.2 --- Function of growth hormone --- p.28
Chapter 2.3.3 --- Cortisol --- p.30
Chapter 2.3.3.1 --- Structure --- p.30
Chapter 2.3.3.2 --- Functions of cortisol --- p.31
Chapter 2.4 --- Sparus sarba --- p.34
Chapter 2.5 --- Urinary bladder of fish --- p.36
Chapter Chapter 3: --- Effect of salinity on the expression of Na+-K+ ATPase and aquaporin-1 in the urinary bladder of silver sea bream Sparus sarba --- p.38
Chapter 3.1 --- Introduction --- p.38
Chapter 3.2 --- Chronic effect of salinity on the expression of Na+-K+ ATPase and aquaporin-1 in the urinary bladder --- p.40
Chapter 3.2.1 --- Materials and Methods --- p.40
Chapter 3.2.1.1 --- Fish --- p.40
Chapter 3.2.1.2 --- Tissue sampling --- p.41
Chapter 3.2.1.3 --- Protein extraction and quantification --- p.41
Chapter 3.2.1.4 --- Na+-K+ ATPase ATPase activity --- p.42
Chapter 3.2.1.5 --- RNA extraction and first strand cDNA synthesis --- p.43
Chapter 3.2.1.6 --- Validation of semi-quantitative RT-PCR --- p.45
Chapter 3.2.1.7 --- Semi-quantification of expression of Na+-K+ ATPase and aquaporin-1 in the urinary bladder --- p.47
Chapter 3.2.1.8 --- Statistical analysis --- p.47
Chapter 3.2.2 --- Results --- p.48
Chapter 3.2.2.1 --- Na+-K+ ATPase activity --- p.48
Chapter 3.2.2.2 --- Relative expression of Na+-K+ ATPase and aquaporin-1 in urinary bladder --- p.48
Chapter 3.2.3 --- Discussion --- p.54
Chapter 3.2.3.1 --- Chronic effect of salinity on Na+-K+ ATPase in urinary bladder --- p.54
Chapter 3.2.3.2 --- Chronic effect of salinity on AQP-1 expression in urinary bladder --- p.59
Chapter 3.3 --- Effect of abrupt transfer on the expression of Na+-K+ ATPase and aquaporin-1 in the urinary bladder --- p.61
Chapter 3.3.1. --- Materials and Methods --- p.61
Chapter 3.3.1.1 --- Fish --- p.61
Chapter 3.3.1.2 --- Tissue sampling --- p.62
Chapter 3.3.1.3 --- "RNA extraction, first strand cDNA synthesis and RT-PCR" --- p.62
Chapter 3.3.1.4 --- Statistical analysis --- p.63
Chapter 3.3.2 --- Results --- p.64
Chapter 3.3.2.1 --- Effect of abrupt hypo-osmotic transfer on the expression of Na+-K+ ATPase and aquaporin-1 in the urinary bladder --- p.64
Chapter 3.3.2.2 --- Effect of abrupt hyper-osmotic transfer on the expression of Na+-K+ ATPase and aquaporin-1 in the urinary bladder --- p.65
Chapter 3.3.3 --- Discussion --- p.73
Chapter 3.4 --- Effect of in vitro salinity on the expression of Na+-K+ ATPase and aquaporin-1 in the urinary bladder --- p.78
Chapter 3.4.1 --- Materials and Methods --- p.78
Chapter 3.4.1.1 --- Fish --- p.78
Chapter 3.4.1.2 --- Tissue sampling --- p.78
Chapter 3.4.1.3 --- Preparation of culture medium --- p.79
Chapter 3.4.1.4 --- "RNA extraction, first strand cDNA synthesis and RT-PCR" --- p.79
Chapter 3.4.1.5 --- Statistical analysis --- p.80
Chapter 3.4.2 --- Results --- p.81
Chapter 3.4.3 --- Discussion --- p.85
Chapter 3.5 --- Conclusion --- p.86
Chapter Chapter 4: --- Effect of hormones on the expression of Na+-K+ ATPase and aquaporin-1 in the urinary bladder of silver sea bream Sparus sarba --- p.88
Chapter 4.1 --- Introduction --- p.88
Chapter 4.2 --- In vivo effect of hormones on the expression of Na+-K+ ATPase and aquaporin-1 in the urinary bladder of silver sea bream Sparus sarba --- p.91
Chapter 4.2.1 --- Material and method --- p.91
Chapter 4.2.1.1 --- Fish --- p.91
Chapter 4.2.1.2 --- Tissue sampling --- p.92
Chapter 4.2.1.3 --- "RNA extraction, first strand cDNA synthesis and RT-PCR" --- p.92
Chapter 4.2.1.4 --- Statistical analysis --- p.92
Chapter 4.2.2 --- Results
Chapter 4.2.2.1 --- Hormonal effect on mRNA expression of Na+-K+ ATPase and AQP-1 in urinary bladder of sea water adapted fish --- p.93
Chapter 4.2.2.2 --- Hormonal effect on mRNA expression of Na+-K+ ATPase and AQP-1 in urinary bladder of brackish water adapted fish --- p.97
Chapter 4.2.3 --- Discussion --- p.101
Chapter 4.2.3.1 --- Effect of prolactin on mRNA expression of Na+-K+ ATPase and AQP-1 in urinary bladder --- p.101
Chapter 4.2.3.2 --- Effect of growth hormone on mRNA expression of Na+-K+ ATPase and AQP-1 in urinary bladder --- p.104
Chapter 4.2.3.3 --- Effect of cortisol on mRNA expression of Na+-K+ ATPase and AQP-1 in urinary bladder --- p.106
Chapter 4.3 --- In vitro effect of hormone on the expression of Na+-K+ ATPase and aquaporin-1 in the urinary bladder of silver sea bream Sparus sarba --- p.109
Chapter 4.3.1 --- Materials and methods --- p.109
Chapter 4.3.1.1 --- Fish --- p.109
Chapter 4.3.1.2 --- Tissue sampling --- p.110
Chapter 4.3.1.3 --- Preparation of culture medium --- p.110
Chapter 4.3.1.4 --- "RNA extraction, first strand cDNA synthesis and RT-PCR" --- p.111
Chapter 4.3.1.5 --- Statistical analysis --- p.111
Chapter 4.3.2 --- Results --- p.112
Chapter 4.3.2.1 --- Effect of prolactin on the expression of Na+-K+ ATPase and aquaporin-1 in the urinary bladder --- p.112
Chapter 4.3.2.2 --- Effect of growth hormone on the expression of Na+-K+ ATPase and aquaporin-1 in the urinary bladder --- p.113
Chapter 4.3.2.3 --- Effect of cortisol on the expression of Na+-K+ ATPase and aquaporin-1 in the urinary bladder --- p.123
Chapter 4.3.3 --- Discussion --- p.124
Chapter 4.3.3.1 --- Effect of prolactin on the expression of Na+-K+ ATPase and aquaporin-1 in the urinary bladder --- p.124
Chapter 4.3.3.2 --- Effect of growth hormone on the expression of Na+-K+ ATPase and aquaporin-1 in the urinary bladder --- p.125
Chapter 4.3.3.3 --- Effect of cortisol on the expression of Na+-K+ ATPase and aquaporin-1 in the urinary bladder --- p.127
Chapter 4.4 --- Conclusion --- p.129
Chapter Chapter 5 --- General Conclusions --- p.131
References --- p.136

All these findings can help us to elucidate the mechanisms for the fish to detect changing osmotic conditions and transform signals to osmoregulatory responses.
In the first part of the study, PRL and PRL-releasing peptide (PrRP) cDNAs have been isolated from euryhaline silver sea bream. The PRL cDNA consists of 1360 bp encoding 212 amino acids whereas the PrRP cDNA contains 631 bp encoding prepro-PrRP with 122 amino acids. PRL mRNA was uniquely expressed in sea bream pituitary but PrRP mRNA was expressed in a variety of tissues. Expression levels of both PRL and PrRP mRNA have been examined in sea bream adapted to different salinities (0, 6, 12, 33 and 50 ppt). In pituitary, both PRL and PrRP mRNA were synchronized in their expression, being significantly higher in fish adapted to low salinities (0 and 6 ppt), but the expression profile of hypothalamic PrRP in different salinities was different. These data suggested that PrRP may possibly act as a local modulator in pituitary rather than a hypothalamic factor for regulating pituitary PRL expression in silver sea bream.
In the second part of the study, silver sea bream abruptly transferred from 33 to 6 ppt exhibited a remarkable pituitary PRL secretion following closely with the temporal changes in serum osmolality and ion levels. In order to investigate the direct effect of extracellular osmolality to pituitary PRL secretion, sea bream pituitary cells were dispersed and exposed to a medium with reduced ion levels and osmolality. PRL released from pituitary cells was found to be significantly elevated. In hyposmotic exposed anterior pituitary cells, cell volume exhibited a 20% increase when exposed to a medium with a 20% decrease in osmolality. These enlarged pituitary cells did not shrink until the surrounding hyposmotic medium was replaced, a phenomenon suggesting an osmosensing ability of silver sea bream PRL cells for PRL secretion in response to a change in extracellular osmotic pressure.
In the third part, olfactory rosette in the nasal cavity was surgically removed from silver sea bream adapted to 6 ppt and 33 ppt and mRNA expression of PRL and PrRP in silver sea bream were measured. The elevated pituitary PRL and PrRP mRNA expression levels as seen in 6 ppt-adapted fish were abolished by this olfactory lamellectomy. On the other hand, hypothalamic PrRP mRNA expression in 6 ppt-adapted fish did not change but those in 33 ppt-adapted fish increase significantly after olfactory lamellectomy. These data suggest a possible osmosensing role of the olfactory system for regulation of PRL expression during hypo-osmotic acclimation of the fish. Besides, calcium-sensing receptor (CaSR) was cloned and its mRNA expression in olfactory system, as shown in other fish species previously, was investigated. However, no CaSR expression could be detected in olfactory rosette and nerve but its expression was demonstrated in osmoregulatory tissues and brain. There was no significant difference in CaSR mRNA expression in pituitary, kidney and anterior intestine of fish adapted to different salinities. These studies could not provide conclusive evidence to correlate CaSR with osmosensing in silver sea bream.
The present study used silver sea bream (Sparus sarba ) as a euryhaline fish model to investigate the regulation of prolactin (PRL) expression and secretion in fish adapted to different salinities.
Kwong, Ka Yee.
Adviser: Norman Y. S. Woo.
Source: Dissertation Abstracts International, Volume: 70-06, Section: B, page: 3248.
Thesis (Ph.D.)--Chinese University of Hong Kong, 2008.
Includes bibliographical references (leaves 154-184).
Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
Abstracts in English and Chinese.
School code: 1307.

碩士
國立臺灣海洋大學
環境生物與漁業科學學系
106
The goldlined seabream (Rhabdosargus sarba) is an important economic species on the western coast of Taiwan, and it is also one of the species for release program in recent years. The main purpose of this study is to examine the reproductive biology of goldlined seabream. A total of 802 specimens (443 females and 232 males) caught by otter trawl, longline, pole and lines boots, bottom gill net or gill net between Jan. 2016 and Dec. 2016 were collected in Penghu fishing port. The relation between body weight (BW) and fork length (FL) was BW=0.0280FL^2.9234 (sexes combined)(n = 675, p<0.05). By observing the tissue sections of the oocyte development, can be divided into eight stages: Chromatin-nucleolus stage, Peri-nucleolus stage, Yolk vesicle stage, pre-yolk stage, post-yolk stage, Migratory nucleus stage, Ripe egg stage and Atretic oocytes stage by the histological examination and ovarian development measurements. By the macroscopic appearance of ovarian, histological examination of ovaries, oocyte diameter measurements, GSI, and monthly percentage in each maturity stage, R. sarba is asynchronous and batch spawner type species, which the spawning season is from December to March in the waters of Penghu. The fecundity of this species ranged from 231,493 to 679,372 eggs, with a mean of 427,166 ± 229,229 eggs. Batch fecundity of this species ranged from 105,872 to 297,557 eggs, with a mean of 179,767 ± 103,103 eggs. Both fecundity and batch fecundity increased with the fork length, body weight, and gonad weight. Size at 50% maturity was estimated to be 23.68 cm FL and 20.89 cm FL for females and males, respectively based on the relation between proportion of maturity and fork length.

Also, the branchial hsp70 levels in fish following chronic salinity acclimation and abrupt hypo-osmotic exposure to 6 ppt were assessed by Western blotting. Upon chronic salinity acclimation, the lowest branchial hsp70 level was found in fish cultured in an iso-osmotic salinity of 12 ppt and the highest was in 50 ppt and 6 ppt environments. Freshwater acclimation resulted in return to lower hsp70 level. The results indicated that iso-osmotic salinity would bring about the least stress level while 50 ppt and 6 ppt were the most stressful salinities to Sparus sarba as indicated by using hsp70 expression as a biomarker of stress. Compared to 50 ppt and 6 ppt, the stress level of fish in fresh water was lower. On the other hand, Sparus sarba exhibited a significant increase in branchial hsp70 level immediately after abrupt hypo-osmotic exposure to 6 ppt when compared with seawater fish sampled at the same time point and increased hsp70 level was sustained throughout the sampling period, indicating the exposure was stressful to the fish.
In the present study, pituitary and serum levels of prolactin in a marine teleost, Sparus sarba, chronically acclimated to various salinities: fresh water (0 ppt), hypo-osmotic (6 ppt), iso-osmotic (12 ppt), normal seawater (33 ppt) and hypersaline (50 ppt) or abruptly exposed to a hypo-osmotic environment of 6 ppt were quantified by the developed peptide-based indirect ELISAs. Progressive increases in pituitary and serum prolactin were found as chronic salinity acclimation progressed from seawater to fresh water. Also, prolactin secretion was immediately induced by abrupt hypo-osmotic exposure to 6 ppt and remained significantly elevated up to 5 days post-exposure to 6 ppt. The results underline the importance of prolactin in marine teleosts kept in fresh water or waters of low salinity. However, there was no significant difference in pituitary prolactin during the course of the abrupt hypo-osmotic exposure experiment. The results may indicate that prolactin might be secreted rapidly from pituitary in large quantities to cope with abrupt exposure to a low-salinity environment.
In the present study, the effects of pharmacological drugs on prolactin levels in pituitary and serum of Sparus sarba were investigated. An increase in prolactin synthesis and release but a decrease in branchial hsp70 expression were found after treatment with sulpiride, a DA-D2 receptor antagonist. In contrast, a reduction in prolactin levels in pituitary and serum but an elevation in hsp70 level in gill were observed following administration of bromocriptine, a DA-D2 receptor agonist. Since hsp70 expression indicates the stress levels, the results of these studies supported the notion that increased prolactin synthesis and release might be related to a reduced stress state and prolactin might have a protective effect on stress tolerance in fish.
Lastly, the role of prolactin in regulating apoptosis in Sparus sarba branchial cells was examined. Successful induction of apoptosis was indicated by an increase in the apoptotic parameter caspase-3 activity in primary cultures of Sparus sarba branchial cells treated with camptothecin, a specific inducer of apoptosis. In this study, prolactin was shown to be anti-apoptotic in Sparus sarba branchial cells as co-treatment with ovine prolactin (oPRL) and camptothecin has been observed to attenuate the elevated caspase-3 activity in gill cell primary cultures. Also, prolactin was found to protect the branchial cells from apoptosis by maintaining the hsp70 level in the cells treated with camptothecin.
The objectives of the present study were to investigate the roles of prolactin in salinity adaptation, hsp70 expression and apoptosis in silver sea bream (Spaurs sarba). Firstly, specific peptide-based indirect ELISAs were developed for pituitary and serum prolactin of Sparus sarba. These assays had been validated by parallelism between the dilution response curves using serially diluted pituitary homogenate and serum sample with the standard curves of the synthetic peptide derived from the amino acid sequence of black sea bream (Acanthopagrus schlegelii ) prolactin.
Ng, Ho Yuen Andus.
"September 2007."
Adviser: N. Y. S. Woo.
Source: Dissertation Abstracts International, Volume: 69-08, Section: B, page: 4567.
Thesis (Ph.D.)--Chinese University of Hong Kong, 2007.
Includes bibliographical references (p. 143-189).
Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
Abstracts in English and Chinese.
School code: 1307.

Luk Chun-yin.
Thesis (M.Phil.)--Chinese University of Hong Kong, 2001.
Includes bibliographical references (leaves 119-131).
Abstracts in English and Chinese.
ABSTRACT --- p.i
ACKNOWLEDGEMENTS --- p.iv
LIST OF FIGURES --- p.x
LIST OF TABLES --- p.xii
Chapter CHAPTER 1 --- GENERAL INTRODUCTION --- p.1
Chapter CHAPTER 2 --- LITERATURE REVIEW --- p.6
Chapter 2.1 --- Introduction --- p.7
Chapter 2.2 --- Ammonia chemistry --- p.10
Chapter 2.3 --- Ammonia metabolism and excretion --- p.11
Chapter 2.3.1 --- Ammonia production --- p.11
Chapter 2.3.2 --- Blood levels of ammonia --- p.12
Chapter 2.3.3 --- Ammonia Excretion --- p.17
Chapter 2.4 --- Urea metabolism and excretion --- p.23
Chapter 2.4.1 --- Urea Chemistry --- p.23
Chapter 2.4.2 --- Urea production in fishes --- p.24
Chapter 2.4.3 --- Argininolysis --- p.25
Chapter 2.4.4 --- Uricolysis --- p.26
Chapter 2.4.5 --- Ornithine-urea Cycle (OUC) --- p.28
Chapter 2.4.5.1 --- Tilapia inhabiting the highly alkaline Lake Magadi --- p.32
Chapter 2.4.5.2 --- High Ambient Ammonia --- p.33
Chapter 2.4.5.3 --- Air Exposure --- p.34
Chapter 2.4.5.4 --- Toadfishes --- p.34
Chapter 2.4.6 --- Blood urea concentration --- p.35
Chapter 2.4.7 --- Urea excretion in fishes --- p.37
Chapter 2.4.7.1 --- Branchial urea excretion in fishes --- p.37
Chapter 2.4.7.2 --- Mechanisms of renal excretion in fishes --- p.40
Chapter 2.5 --- Influence of environmental salinity on nitrogen excretion in teleosts --- p.42
Chapter CHAPTER 3 --- BODY COMPOSITION AND UREA BIOSYNTHESIS OF SPAR US SARBA IN DIFFERENT SALINITIES --- p.46
Chapter 3.1 --- Introduction --- p.47
Chapter 3.2 --- Materials and Methods --- p.49
Chapter 3.2.1 --- Experimental animals --- p.49
Chapter 3.2.2 --- Tissue sampling --- p.49
Chapter 3.2.3 --- Water chemistry analysis --- p.50
Chapter 3.2.4 --- Hematological parameters --- p.50
Chapter 3.2.5 --- Metabolite and electrolyte contents --- p.51
Chapter 3.2.6 --- Hepatic enzymes activities --- p.51
Chapter 3.2.6.1 --- Tissue preparation --- p.51
Chapter 3.2.6.2 --- Carbamyl phosphate synthetases (CPSases; E.C. 2.7.2.5) --- p.52
Chapter 3.2.6.3 --- Ornithine carbamoyl transferase (OCTase; E.C. 2.1.3.3) --- p.53
Chapter 3.2.6.4 --- Argininosuccinate synthetase (ASS; E.C. 6.3.4.5) --- p.54
Chapter 3.2.6.5 --- Argininosuccinate lyase (ASL; E.C. 4.3.2.1) --- p.54
Chapter 3.2.6.6 --- Arginase (ARG; 3.5.3.1) --- p.55
Chapter 3.2.6.7 --- Glutamate dehydrogenase (EC 1.4.1.3) --- p.55
Chapter 3.2.6.8 --- Uricase (E.C. 1.7.3.3) --- p.56
Chapter 3.2.6.9 --- Allantoinase --- p.57
Chapter 3.2.6.10 --- Allantoicase --- p.57
Chapter 3.2.7 --- Statistical analysis --- p.58
Chapter 3.3 --- Results --- p.59
Chapter 3.3.1 --- "Changes in hepatosmatic index, renal somatic index, muscle water and lipid content and hematological parametersin response to different salinity acclimation" --- p.59
Chapter 3.3.2 --- Changes in serum chemistry in response to different salinity acclimation --- p.60
Chapter 3.3.3 --- Changes in hepatic ornithine-urea cycle enzyme activitiesin response to different salinity acclimation --- p.61
Chapter 3.3.4 --- Changes in GDHase and uricolytic enzyme activitiesin response to different salinity acclimation --- p.62
Chapter 3.4 --- Discussion --- p.71
Chapter 3.4.1 --- Hematological responses --- p.72
Chapter 3.4.2 --- Muscle moisture content --- p.74
Chapter 3.4.3 --- Circulating electrolyte levels --- p.75
Chapter 3.4.4 --- Circulating metabolites levels --- p.77
Chapter 3.4.5 --- Urea metabolism --- p.80
Chapter 3.4.5.1 --- Ornithine-urea cycle enzymes --- p.80
Chapter 3.4.5.2 --- Carbamoyl phosphate synthetase isozymes --- p.81
Chapter 3.4.5.3 --- Uricolytic pathway and argininolysis --- p.85
Chapter 3.4.5.4 --- Influence of salinity on urea metabolism --- p.86
Chapter 3.4.6 --- Conclusion --- p.87
Chapter CHAPTER 4 --- EFFECT OF SALINITY ON NITROGEN EXCRETION OF SPARUS SARBA --- p.88
Chapter 4.1 --- Introduction --- p.89
Chapter 4.2 --- Materials and Methods --- p.91
Chapter 4.2.1 --- Experimental animals --- p.91
Chapter 4.2.2 --- Experimental protocol --- p.92
Chapter 4.2.3 --- Determination of net ammonia and urea excretion rates --- p.94
Chapter 4.2.4 --- Statistical analysis --- p.94
Chapter 4.3 --- Results --- p.95
Chapter 4.3.1 --- Net ammonia-N and urea-N excretion rates --- p.95
Chapter 4.3.2 --- Changes in net ammonia-N and urea-N excretion ratesin response to abrupt hyposmotic exposure --- p.95
Chapter 4.3.3 --- Changes in net ammonia-N and urea-N excretion rates after exposure to amiloride for 3 hours --- p.96
Chapter 4.3.4 --- Changes in net urea-N excretion rates in response to elevated body urea levels --- p.96
Chapter 4.3.5 --- Changes in net ammonia-N excretion rates in response to elevated body ammonia levels --- p.97
Chapter 4.4 --- Discussion --- p.106
Chapter 4.4.1 --- Influence of environmental salinity on net ammonia-N and urea-N excretion rates --- p.106
Chapter 4.4.2 --- Effects of amiloride on nitrogen excretion --- p.109
Chapter 4.4.3 --- Effect of increased body ammonia on ammonia excretion --- p.113
Chapter 4.4.4 --- Changes in net urea-N excretion rates in response to elevated body urea levels --- p.113
Chapter 4.5 --- Conclusion --- p.114
Chapter CHAPTER 5 --- GENERAL CONCLUSION --- p.115
references --- p.119

碩士
國立海洋大學
水產養殖學系
88
英文摘要 The objective of the present study is to produce fast growing sliver sea bream (Sparus sarba) for intensive aquaculture by employing various mass gene transfer technologies. We have transferred a "all-fish" growth hormone (GH) gene construct into silver sea bream embryos by electroporating the transgene construct into sperm. Ranging from 50 to 70% of the survived animals developed from eggs fertilized with electroporated sperm were found PCR positive. Striping gametes from sliver sea bream broods can cause severe stress to fish, frequently leading to serious mortality, and most female broods can only be striped once. To eliminate handling stresses in the brood fish, an alternative mass gene transfer method is developed. We employed lipid vesicles made of lipospermine L-dioleoyl phosphatidylethanolamine (DOPE) to deliver the GH transgene into testis. Each male fish received 0.7 ml of liposome/DNA mixture. Forty-eight hours after gonadal lipofection, treated male fish were mated to untreated females and fertilized eggs were collected every day. Ranging from 17% to 50% of the resulting larvae were tested positive for the transgene by PCR analysis. The percentage (58-90%) of PCR positive fry was significantly improved if multiple gonadal lipofection was carried out in the same male fish. Furthermore, preliminary analysis showed that a significant number of PCR positive GH transgenic silver sea bream can promote the sea bream growth. Body component analysis of transgenic fish indicated that the fat content of transgenic sea bream muscle was lower than the control sea bream in muscle tissue. Transgene expression also has been demonstrated in PCR positive fish by RT-PCR analysis. These results suggest that faster growing silver sea bream can be produced by various mass gene transfer technologies.

碩士
中興大學
分子生物學研究所
95
Gonadotropins, including follicle-stimulating hormone (FSH) and luteinizing hormone (LH), are the key regulators of vertebrate reproduction, and they are synthesized and secreted from the gonadotrophs in the anterior pituitary. FSH and LH exert their effects on ovarian or testicular functions through the membrane receptors (FSH receptor, FSHR; LH receptor, LHR) on the gonadal cells. Most studies on the reproduction of mammals and fish, but scant about of commercial fish – silver sea bream (Sparus sarba) in Taiwan. Here we report on the first cloning of full length FSHR and LHR sequences of silver sea bream, and on the characterization and expression of these two genes. Besides, the effect of spawning was also examined. Silver sea bream FSHR and LHR have been cloned by 5´RACE and 3´RACE, and their expression was analysis by semi-quantitative RT-PCR, RNA in situ hybridization and immunohistochemistry. The cloned FSHR cDNA sequence has 2,402 bp, which composed of a coding region of 704 a.a. (GenBank accession no. DQ522160), and LHR cDNA sequence has 2,514 bp, whose coding region has 691 a.a. (GenBank accession no. DQ522161). Amino acid sequences of the coding region of silver sea bream FSHR has a 51.2% identities with that of mammals, 50.5% with bird, 69.5% with fish, 50.6% with amphibian and 48.5% with reptiles; and amino acid sequence of the coding region of LHR has a 50.7% identities with that of mammals, 52.1% with bird and 59.5% with fish. The expression of FSHR 2.1 kb fragment in nonspawning (April to November) and spawning (December to next March) seasons was detected by RT-PCR, which was mainly expressed in the brain and gonads of both genders; the expression level in testes in spawning season was 1.1 fold higher than that in nonspawning season, but no difference existed between those of ovaries in the seasons. LHR 2.0 kb fragment in nonspawning season of male silver sea bream was detected in brain, liver and testis, while in brain, intestines, kidney and ovary in female, in spawning season male silver sea bream detected in brain, heart, liver and testis, while in brain, heart and ovary in female. The LHR expression level in testes and ovaries in spawning season was 0.7 and 1.8 fold, respectively, higher than that in nonspawning season. These data indicated that the expression of gonadotropin receptors could be affected by seasons. Expression of the FSHR and LHR RNA and proteins were also detected by in situ hybridization and immunohistochemistry in the tissues mentioned before. These results are consistent with those analyzed by RT-PCR. These basic data could be applied in the studies of reproductive regulation and dysfunction, and of biomedicine, and further, it is also an alternative in the development of biocontrol of transgenic animals.

Li Jun.
"March 2002."
Thesis (Ph.D.)--Chinese University of Hong Kong, 2002.
Includes bibliographical references (p. 189-216).
Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
Mode of access: World Wide Web.
Abstracts in English and Chinese.

The present study provided information for the role of the RAS in seabream osmoregulatory responses. The structure of angiotensinogen suggested that flounder type Ang II was the prevalent form in seabream. However, HPLC analysis suggested that different forms of angiotensins were present in seabream adapted to different salinities. The status of RAS was revealed in seabream adapted to different salinities and a higher status was found in hypersaline environment. Local renal RAS was identified and it may be activated in hyposmotic media and associated with an increase in glomerular and tubular function to excrete excess water. In general, the RAS in seabream displays differential status, both at systemic and local levels, which modulates osmoregulatory functions under acute and chronic salinity perturbation.
The renin angiotensin system (RAS) is involved in the control of body fluid homeostasis in silver seabream. Seabream angiotensinogen was cloned and sequenced in the present study. The sequence alignment showed that the angiotensinogen of seabream is most similar to that of pufferfish. Differential status of RAS was found among different salinities, with relatively higher RAS activity among hyperosmotic adapted seabream. Circulating angiotensin II (Ang II) was higher in hyperosmotic adapted seabream, with the highest value observed in seabream adapted to double-strength seawater. Although the level of immunoreactive angiotensins in freshwater adapted seabream was higher than that of brackish-water, Ang III, but not Ang II, was the prevalent circulating form in freshwater adapted seabream according to HPLC analysis. Hepatic angiotensinogen expression, however, did not show any statistical difference among different salinities. A positive feedback control for angiotensinogen by Ang II is present in the hepatic tissue of seabream as Ang II increased the expression of angiotensinogen in isolated hepatocyte but captopril lowered the angiotensinogen expression in intact fish. Branchial Na-K-ATPase activities were elevated by Ang II and the activities among different salinities showed a pattern similar to that of circulating angiotensins. However, upon abrupt hyposmotic transfer, branchial Na-K-ATPase elevated along with a decrease in circulating Ang II, an observation implying that the relationship between Na-K-ATPase and Ang II may only be causal. Captopril blockade not only lowered not only circulating Ang II levels but also that of cortisol, indicating RAS activity may limit cortisol secretion. An elevation in the circulating cortisol may be related to the increase in branchial Na-K-ATPase activities after abrupt hyposmotic transfer. The stimulatory effect on branchial Na-K-ATPase activity and the vasopressor effect of Ang II were more potent in hyposmotic than hyperosmotic adapted seabream, which indicates hyposmotic adapted seabream is more sensitive to RAS activation. The renal RAS in silver seabream functions independently from the systemic RAS as the pattern of renal angiotensins was dissimilar to that of systemic angiotensins. The renal RAS was activated in brackish water conditions and abrupt hyposmotic transfer significantly increased renal RAS activities. Kidney morphometrics also indicated that hyposmotic adaptation increase the filtering capacity of seabream nephrons. The number and diameter of glomeruli increase significantly in freshwater adapted seabream, which may vastly increase the filtering surface of the nephrons. Collecting tubules were more prevalent in the kidney of hyposmotic adapted seabream, with higher number, diameter and thickness, suggesting a lower water permeability of collecting tubules is essential for the formation of copious and diluted urine in hyposmotic environment.
Wong Kwok Shing.
"December 2005."
Adviser: Norman Y. S. Woo.
Source: Dissertation Abstracts International, Volume: 67-11, Section: B, page: 6144.
Thesis (Ph.D.)--Chinese University of Hong Kong, 2005.
Includes bibliographical references (p. 130-145).
Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
Abstracts in English and Chinese.
School code: 1307.

碩士
國立臺灣海洋大學
環境生物與漁業科學學系
107
The goldlined seabream (Rhabdosargus sarba) is a common species caught by the Taiwanese coastal fisheries. However, its biological information on spawning ground, migration route, reproductive biology, and age and growth is little known. In total, 593 specimens ranged from 12.3 to 44.0 cm in total length (TL) and 28.9 to 1832.0 g in body weight (BW) were collected from the coastal waters off Penghu from September 2015 to August 2016. The maximum likelihood ratio test indicated the sex-specific relationship between body weight and length is not significantly different and can be expressed as: BW= 0.01511 TL3.0346 (r2= 0.950; n= 593). The edge analysis indicated the growth rings on otolith were formed once per year, and the opaque zones were deposited in December. Age estimate was made by counting opaque zones in sagittal otolith sections of 447 specimens under transmitted light. Three growth functions namely the VBGF, the Robertson and the Gompertz function were used in this study. Based on AICc and w_i, VBGF was chosen to be the best growth function for R. sarba. The maximum likelihood ratio test indicated that no significant difference on growth between females and males and the parameters of VBGF for R. sarba (sexes combined) were L∞=53.94 cm TL, k=0.217 year-1, t0=-0.182 year. The oldest ages were 8.17 and 8.00 years old for females and males, respectively.

碩士
國立海洋大學
漁業研究所
77
1.By the marphological and ecological characteristics, the development it larval Sparus sarba were divided into thrse stages: (1)Prelarvae stage(TL:3-4mm)(2)Postlarvae stage(TL:4-11mm) (3)Juvenils stage(TL:11-25mm) 2.The survival rate of the starved larvae was decreased with age after their mouths opened. All larvae died at the 9th day, and their PHK(point of no return) was between the 6th and the 7th day. Loconotion duration of the fed and unfed groups began to show the differences at the end of yolk-sac stage. Total lengths of these two groups showed significant differences after their oil gloubles were absorbed. Start feeding at the 3rd day had the highest survival rate. 3.Before the netanorphosis it was pure-cone retina. (Rods were not identified until the 40th day). The larvae showed negative phototactis at 28 days old. The density ratio of total photoreceptor nuclei to the cone numbers in the was greater than 1 at the age of 35 days old. It is likely that the rods appeared before metamorphosis. The newly hatched larvae have unpigmented and presumably functionlass retinae. On the 3rd day there was sparse pigment distributed. The retinomotor responses of the pigment aigration was synchronized with the period of their netanorphosis. The acuity of larvae depended on the focal length (which increases linearly with age) and on the density of cones. During the metamorphosis, the resolving angle decrease significantly. As the larvae grew, the acuity improved. 4.The effects of light intensity on feeding behavior as follows: (1)The higher of light intensity, the higher feeding rations in larvae and juveniles, (2)Under the natural photoperiod, continuous illumination and reverse photoperiod (illumination at night). The juveniles(TL:12-15 mm) fed artenias, the young (TL:30- 40mm) fed shriap meat. Both of their feeding amount were higher in light-period, and the feeding period was changed by light. It indicated that the larvae was "visual feeder", and the activity pattern was diurnal. (3)The growth rate reared in long photoperiods (L/D=16/8) was greater than those in the short photoperiods(L/D). (4)The juveniles(TL:12-15mm) under dark adaption, feeding artemias by chance and the young(TL:28-39mm) feeding without vision was not by chance alone.

催乳素是一種負責廣鹽性硬骨魚低滲環境適應的重要激素。最近發現的催乳素釋放肽被報導在魚類生理活動中參與了刺激催乳素釋放和滲透調節過程。本研究以黃錫鯛為實驗動物，旨在詮釋催乳素和催乳素釋放肽在廣鹽性硬骨魚中的滲透調節作用和調控機制。
實驗第一部份將黃錫鯛催乳素在大腸桿菌中過量表達並用鎳離子親和層析法將之純化。此重組黃錫鯛催乳素可被抗黃錫鯛催乳素抗體特異識別。將此重組催乳素注射入活體紅箭魚可導致其血漿鈉離子濃度和滲透壓顯著提高，從而證明了此催乳素的生物活性。在第二部份的研究中，黃錫鯛催乳素受體和催乳素釋放肽受體的全長編碼基因被克隆和分析。黃錫鯛催乳素受體和硬骨魚二型催乳素受體蛋白質相似性較高（可達47%至94%），但同魚類一型催乳素受體及其他高等脊椎動物催乳素受體同源性較低，相似性只有22%至37%。此催乳素受體廣泛表達在各種組織器官中，包括腦組織、腮、心臟、腸、腎、肌肉及性腺。黃錫鯛催乳素釋放肽受體屬於G-蛋白偶聯受體家族，它具備該家族成員的各種保守序列特徵。並且它含有同其他催乳素釋放肽受體一樣的PDZ域結合位點。組織表達分佈結果顯示此受體在中樞神經系統各部份及腮、心臟、肝臟和性腺中有表達。本研究最後一部份以黃錫鯛原代培養腮細胞為實驗對象，分別探究了腮細胞對於不同滲透壓力、催乳素和催乳素釋放肽處理的反應。在這些實驗中，我們檢測了腮細胞的死亡及凋亡狀況，并用實時定量PCR分析了一些滲透調節相關基因的表達狀況。另外，我們還採用了蛋白免疫印跡法檢測了腮細胞處於不同滲透壓條件下，其熱休克蛋白70的蛋白質豐度變化。我們的結果顯示：低滲壓力會引起腮細胞死亡包括誘導凋亡。催乳素和催乳素釋放肽處理均可以顯著降低低滲壓力所誘導的細胞死亡，並且催乳素還可以阻止一部份細胞進行凋亡。腮細胞中鈉鉀泵兩個亞基、熱休克同族蛋白70及催乳素受體的基因表達均被催乳素和催乳素釋放肽處理上調。但是水通道蛋白3的基因表達并未出現明顯的變化。當腮細胞暴露于高滲透壓力環境時，水通道蛋白3、囊性纤维化跨膜转运调节因子、鈉鉀泵兩個亞基和熱休克同族蛋白70的基因表達都明顯上調。與之相反，這些基因的表達在處於低滲環境的腮細胞中均被下調。
本研究提供了許多關於催乳素和催乳素釋放肽在魚類腮滲透調節過程中的詳細作用，并且首次報導了催乳素釋放肽可以直接作用于魚類滲透調節組織發揮功能從而參與滲透調節過程，而不只是如以往被認為的通過旁分泌或者自分泌方式刺激催乳素的表達來發揮生理功能。本研究的探索和發現使我們更加深刻地理解催乳素和催乳素釋放肽在廣鹽性硬骨魚鹽度適應過程中的調控機制。
Prolactin (PRL) is well known as a crucial hormone responsible for fresh (hypoosmotic) water acclimation in euryhaline teleosts. The recently discovered prolactin-releasing peptide (PrRP) has been reported to take part in stimulating PRL release and affecting osmoregulatory processes in fish. This study aims to investigate the osmoregulatory effects and regulatory mechanisms of PRL and PrRP in a euryhaline teleost, silver sea bream (Sparus sarba).
First, silver sea bream prolactin (ssPRL) was over-expressed in E. coli by IPTG induction and purified by Ni-based immobilized metal ion affinity chromatography. This recombinant silver sea bream prolactin (rssPRL) was recognized by specific antibodies against ssPRL. Subsequently, its bioactivity was confirmed by in vivo injection to swordtails, which resulted in significant increase in plasma Na⁺ level and osmolality. In the second part, full-length cDNAs of silver sea bream PRL receptor (ssPRLR) and PrRP receptor (ssPrRPR) were cloned and characterized. ssPRLR shares high amino acid identities (47% to 94%) with teleost PRLR2s but low identities (22% to 37%) with piscine PRLR1s and higher vertebrate PRLRs. It is widely distributed in brain, gill, heart, gut, kidney, muscle and gonad. ssPrRPR belongs to G protein-coupled receptor (GPCR) family with all the conserved features of GPCRs, and possesses the special PDZ domain-binding motifs of other PrRPRs. Its expression was detected in the central nervous system, gill, heart, liver and gonad. Lastly, a primary gill cell culture of silver sea bream was developed and used as a tool for studying responses of gill cells following exposure to media of different osmotic stress, rssPRL and ssPrRP, respectively. Then, cytotoxicity and apoptosis assays were performed, and effects of these treatments on expression profiles of osmoregulatory genes were analyzed by real time PCR. Influence of osmotic stresses on protein abundance of heat shock protein 70s was examined by Western blot. The results showed that hypoosmotic challenge could induce cell death including apoptosis. Both PRL and PrRP treatment markedly decreased the induced cell death, and PRL treatment prevented some gill cells from apoptosis. Expression levels of Na⁺-K⁺-ATPase alpha (NKA-α) and beta (NKA-β) subunits, heat shock cognate 70 (HSC70) and PRLR were up-regulated in PRL- and PrRP- treated gill cells, however no significant effect on expression of aquaporin3 (AQP3) mRNA was apparent. After hyperosmotic exposure, expression levels of AQP3, cystic fibrosis transmembrane conductance regulator (CFTR), NKA-α, NKA-β, and HSC70 were significantly increased. In contrast, hypoosmotic exposure considerably down-regulated expression levels of these genes.
The present study represented considerable addition of details about the precise actions of PRL and PrRP in branchial osmoregulation. Furthermore, it first reported that PrRP can exert direct effects on fish osmoregulatory epithelia, instead of its oft presumed autocrine or paracrine action. All these efforts provide new insights into the control mechanisms of PRL and PrRP in euryhaline teleost during salinity acclimation.
Detailed summary in vernacular field only.
Detailed summary in vernacular field only.
Detailed summary in vernacular field only.
Qu, Zhe.
Thesis (M.Phil.)--Chinese University of Hong Kong, 2012.
Includes bibliographical references (leaves 108-142).
Abstracts also Chinese.
Title page --- p.i
Thesis committee --- p.ii
Acknowledgement --- p.iii
Abstract --- p.v
Abstract-Chinese version --- p.viii
Table of contents --- p.x
List of figures --- p.xiv
List of abbreviations --- p.xvi
Chapter Chapter 1 --- General introduction --- p.1
Chapter Chapter 2 --- Literature review --- p.6
Chapter 2.1 --- The role of gill in teleost osmoregulation --- p.7
Chapter 2.1.1 --- An overview of fish osmoregulation --- p.7
Chapter 2.1.2 --- The gill structure and epithelial ionocytes --- p.8
Chapter 2.1.3 --- Osmoregulatory molecules in gill cells --- p.10
Chapter 2.2 --- Piscine prolactin --- p.16
Chapter 2.2.1 --- General features and distribution --- p.16
Chapter 2.2.2 --- Prolactin receptor --- p.17
Chapter 2.2.3 --- Prolactin and teleost osmoregulation --- p.18
Chapter 2.3 --- Prolactin-releasing peptide in teleost --- p.21
Chapter 2.3.1 --- Discovery and characteristics --- p.21
Chapter 2.3.2 --- Distribution and its receptor --- p.22
Chapter 2.3.3 --- Functions of PrRP --- p.23
Chapter A. --- PrRP and prolactin release --- p.23
Chapter B. --- PrRP and neuroendocrine stress response --- p.24
Chapter C. --- Effects on food intake and energy metabolism --- p.26
Chapter D. --- PrRP and cardiovascular system --- p.27
Chapter Chapter 3 --- Production and characterization of recombinant silver sea bream (Sparus sarba) prolactin --- p.29
Chapter Abstract --- p.30
Chapter 3.1 --- Introduction --- p.31
Chapter 3.2 --- Materials and methods --- p.33
Chapter 3.2.1 --- Experimental animals --- p.33
Chapter 3.2.2 --- Construction of recombinant silver sea bream prolactin expression vector --- p.33
Chapter 3.2.3 --- Large-scale production and purification of rssPRL --- p.34
Chapter 3.2.4 --- SDS polyacrylamide gel electrophoresis and Western blotting analysis of rssPRL --- p.35
Chapter 3.2.5 --- In vivo bioassay of rssPRL --- p.36
Chapter 3.2.6 --- Statistical analysis --- p.37
Chapter 3.3 --- Results --- p.38
Chapter 3.3.1 --- Overexpression and purification of rssPRL --- p.38
Chapter 3.3.2 --- Biological activity assay of rssPRL in vivo --- p.38
Chapter 3.4 --- Discussion --- p.43
Chapter Chapter 4 --- Molecular cloning and characterization of prolactin receptor (PRLR) and prolactin-releasing peptide receptor (PrRPR) in silver sea bream (Sparus sarba) --- p.46
Chapter Abstract --- p.47
Chapter 4.1 --- Introduction --- p.48
Chapter 4.2 --- Materials and methods --- p.51
Chapter 4.2.1 --- Fish and tissue samples --- p.51
Chapter 4.2.2 --- Molecular cloning of full-length ssPRLR and ssPrRPR cDNA --- p.51
Chapter 4.2.3 --- Multiple sequence alignments and phylogenetic analysis --- p.53
Chapter 4.2.4 --- Tissue distribution of ssPRLR and ssPrRPR --- p.53
Chapter 4.3 --- Results --- p.54
Chapter 4.3.1 --- Cloning and characterization of silver sea bream PRLR and PrRPR cDNA --- p.54
Chapter 4.3.2 --- Tissue distribution of ssPRLR and ssPrRPR mRNA --- p.56
Chapter 4.4 --- Discussion --- p.65
Chapter Chapter 5 --- Influences of osmotic stress and hormones on gill cell of silver sea bream (Sparus sarba) --- p.69
Chapter Abstract --- p.70
Chapter 5.1 --- Introduction --- p.72
Chapter 5.2 --- Methods and materials --- p.77
Chapter 5.2.1 --- Fish and primary gill cell culture --- p.77
Chapter 5.2.2 --- Experiments using gill cell culture --- p.78
Chapter 5.2.3 --- Cell survival assay and apoptosis relevant detections --- p.79
Chapter 5.2.4 --- Real time PCR analysis --- p.80
Chapter 5.2.5 --- Western blot analysis of HSP70 --- p.82
Chapter 5.2.6 --- Data processing and statistical analysis --- p.82
Chapter 5.3 --- Results --- p.83
Chapter 5.3.1 --- Hypoosmotic stress induced cell death --- p.83
Chapter 5.3.2 --- Influences of PRL and PrRP on hypoosmotic challenge induced cell death --- p.84
Chapter 5.3.3 --- Effects of hormones and osmotic stresses on expression levels of branchial genes --- p.84
Chapter 5.3.4 --- Osmotic stresses induced changes in branchial protein amount of HSP70s --- p.85
Chapter 5.4 --- Discussion --- p.94
Chapter Chapter 6 --- General discussion and conclusions --- p.102
References --- p.108

Ng Ho Yuen Andus.
Thesis (M.Phil.)--Chinese University of Hong Kong, 2001.
Includes bibliographical references (leaves 105-131).
Abstracts in English and Chinese.
Chapter I --- Title page --- p.i
Chapter II --- Thesis committee --- p.ii
Chapter III --- Acknowledgement --- p.iii
Chapter IV --- Abstract --- p.v
Chapter V --- Abstract (Chinese version) --- p.vii
Chapter V --- Table of contents --- p.ix
Chapter VI --- List of abbreviations --- p.xv
Chapter VII --- List of figures --- p.xviii
General introduction --- p.1
Chapter Chapter 1: --- Literature review --- p.5
Chapter 1.1. --- Heat shock proteins (HSPs) --- p.6
Chapter 1.1.1. --- Introduction --- p.6
Chapter 1.1.2. --- The various heat shock proteins --- p.8
Chapter 1.1.2.1. --- HSP100s --- p.8
Chapter 1.1.2.2. --- HSP90s --- p.9
Chapter 1.1.2.3. --- HSP70s --- p.12
Chapter 1.1.2.3.1. --- ATPase reaction cycle of HSP70 and protein folding --- p.13
Chapter 1.1.2.3.2. --- Protein translocation --- p.14
Chapter 1.1.2.3.3. --- Selective lysosomal proteolysis --- p.16
Chapter 1.1.2.4. --- HSP60s --- p.16
Chapter 1.1.2.5. --- Small HSPs --- p.17
Chapter 1.1.2.6. --- Ubiquitin --- p.19
Chapter 1.1.3. --- HSP studies in fish --- p.21
Chapter 1.1.3.1. --- In vivo works --- p.21
Chapter 1.1.3.2. --- In vitro works --- p.23
Chapter 1.2. --- Growth hormone / prolactin family in teleostean fishes --- p.26
Chapter 1.2.1. --- Introduction --- p.26
Chapter 1.2.2. --- Growth hormone (GH; somatotropin) --- p.29
Chapter 1.2.2.1. --- Structure --- p.29
Chapter 1.2.2.2. --- Actions --- p.29
Chapter 1.2.2.3. --- Insulin-like Growth Factors (IGFs; somatomedins) --- p.31
Chapter 1.2.3. --- Prolactin (PRL) --- p.34
Chapter 1.2.3.1. --- Structure --- p.34
Chapter 1.2.3.2. --- Actions --- p.35
Chapter 1.2.4. --- Somatolactin (SL) --- p.37
Chapter 1.2.4.1. --- Structure --- p.37
Chapter 1.2.4.2. --- Actions --- p.38
Chapter 1.2.5. --- Growth hormone receptor (GH-R) and prolactin receptor (PRL-R) --- p.39
Chapter 1.3. --- Cortisol in teleostean fishes --- p.41
Chapter 1.4. --- Salinity adaptation in teleosts --- p.44
Chapter Chapter 2: --- Effect of in vitro thermal shock on HSP70 expression in whole blood of Sparus sarba --- p.46
Chapter 2.1. --- Introduction --- p.47
Chapter 2.2. --- Materials and methods --- p.49
Chapter 2.2.1. --- Overall experimental design --- p.49
Chapter 2.2.2. --- Experimental fish --- p.49
Chapter 2.2.3. --- Blood sampling and preparation --- p.49
Chapter 2.2.4. --- Thermal stress regimes --- p.50
Chapter 2.2.5. --- Protein extraction --- p.51
Chapter 2.2.6. --- Protein quantification --- p.51
Chapter 2.2.7. --- Indirect enzyme-linked immunosorbent assay (ELISA) --- p.52
Chapter 2.2.8. --- Protein gel electrophoresis and immunoblotting (Western blotting) --- p.54
Chapter 2.2.9. --- Statistical analysis --- p.55
Chapter 2.3. --- Results --- p.56
Chapter 2.3.1. --- Validation of indirect ELISA --- p.56
Chapter 2.3.2. --- Effect of in vitro thermal shock on HSP70 expression in whole blood of Sparus sarba --- p.56
Chapter 2.4. --- Discussion --- p.60
Chapter 2.5. --- Conclusion --- p.64
Chapter Chapter 3: --- Effects of hormones on HSP70 expression in whole blood of Sparus sarba in vitro --- p.65
Chapter 3.1. --- Introduction --- p.66
Chapter 3.2. --- Materials and methods --- p.68
Chapter 3.2.1. --- Overall experimental design and experimental fish --- p.68
Chapter 3.2.2. --- Hormone treatments --- p.59
Chapter 3.2.3. --- "Protein extraction and quantification, indirect ELISA，gel electrophoresis, and immunoblotting (Western blotting)" --- p.70
Chapter 3.2.4. --- Statistical analysis --- p.70
Chapter 3.3. --- Results --- p.71
Chapter 3.3.1. --- Effect of Cortisol on HSP70 levels in whole Blood --- p.71
Chapter 3.3.2. --- Effect of recombinant bream growth hormone on HSP70 levels in whole blood --- p.71
Chapter 3.3.3. --- Effect of recombinant bream insulin-like growth factor-I on HSP70 levels in whole blood --- p.71
Chapter 3.3.4. --- Effect of ovine prolactin on HSP70 levels in whole blood --- p.72
Chapter 3.4. --- Discussion --- p.81
Chapter 3.4.1. --- Effect of Cortisol on HSP70 levels in whole Blood --- p.81
Chapter 3.4.2. --- Effect of recombinant bream growth hormone on HSP70 levels in whole blood --- p.83
Chapter 3.4.3. --- Effect of recombinant bream insulin-like growth factor-I on HSP70 levels in whole blood --- p.85
Chapter 3.4.4. --- Effect of ovine prolactin on HSP70 levels in whole blood --- p.86
Chapter 3.5. --- Conclusion --- p.88
Chapter Chapter 4: --- Effect on HSP70 expression in whole blood of Sparus sarba acclimated to various salinities --- p.89
Chapter 4.1. --- Introduction --- p.90
Chapter 4.2. --- Materials and methods --- p.92
Chapter 4.2.1. --- Overall experimental design and experimental fish --- p.92
Chapter 4.2.2. --- "Protein extraction and quantification, indirect ELISA, gel electrophoresis, and immunoblotting (Western blotting)" --- p.92
Chapter 4.2.3. --- Statistical analysis --- p.93
Chapter 4.3. --- Results --- p.94
Chapter 4.4. --- Discussion --- p.97
Chapter 4.5. --- Conclusion --- p.100
Chapter Chapter 5: --- General discussion and conclusion --- p.101
References --- p.105

Leung Ling Yan.
Thesis (M.Phil.)--Chinese University of Hong Kong, 2004.
Includes bibliographical references (leaves 117-138).
Abstracts in English and Chinese.
Abstract (English) --- p.ii
Abstract (Chinese) --- p.iv
Acknowledgements --- p.vi
List of Abbreviation --- p.vii
List of Figures and Tables --- p.viii
Chapter Chapter 1 --- General Introduction --- p.1
Chapter Chapter 2 --- The effect of salinity on pentose phosphate pathway activity and red blood cells resistance to oxidative stress in silver seabream (Sparus sarba) --- p.4
Abstract --- p.7
Chapter 2.1 --- Literature review --- p.8
Chapter 2.2 --- Materials and Methods --- p.17
Chapter 2.3 --- Results --- p.32
Chapter 2.4 --- Discussion --- p.54
Chapter 2.5 --- Conclusion --- p.63
Chapter Chapter 3 --- The effect of dietary carbohydrate level on pentose phosphate pathway activity and red blood cells resistance to oxidative stress in of silver seabream (Sparus sarba) --- p.65
Abstract --- p.67
Chapter 3.1 --- Literature review --- p.68
Chapter 3.2 --- Materials and Methods --- p.77
Chapter 3.3 --- Results --- p.80
Chapter 3.4 --- Discussion --- p.101
Chapter 3.5 --- Conclusion --- p.112
Chapter Chapter 4 --- Summary --- p.113
References --- p.117

Hui Fong Fong Liza.
Thesis submitted in: December 2002.
Thesis (M.Phil.)--Chinese University of Hong Kong, 2003.
Includes bibliographical references (leaves 130-182).
Abstracts in English and Chinese.
Chapter I --- Title page --- p.I
Chapter II --- Thesis committee --- p.II
Chapter III --- Acknowledgements --- p.III
Chapter IV --- Abstract (Chinese version) --- p.IV
Chapter V --- Abstract (English version) --- p.VII
Chapter VI --- Table of contents --- p.X
Chapter VII --- List of figures --- p.XIV
Chapter VIII --- List of table --- p.XVIII
Chapter Chapter 1: --- General introduction --- p.1
Chapter Chapter 2: --- Literature review --- p.5
Chapter 2.1. --- Gill --- p.5
Chapter 2.2. --- Chloride cells (Mitochondria-rich cells) --- p.6
Chapter 2.2.1. --- Ion extrusion by fish in seawater --- p.9
Chapter 2.2.2. --- Ion uptake by fish in hypo-osmotic condition --- p.12
Chapter 2.3. --- Sparus sarba (Silver seabream) --- p.14
Chapter 2.4. --- Sodium-potassium adenosinetriphosphatase (Na+-K+-ATPase) --- p.15
Chapter 2.4.1. --- Na+-K+-ATPase α-subunit --- p.17
Chapter 2.4.2. --- Na+-K+-ATPase β-subunit --- p.18
Chapter 2.4.3. --- Regulation of Na+-K+-ATPase --- p.20
Chapter 2.5. --- Hormones --- p.21
Chapter 2.5.1. --- Growth hormone-prolactin family --- p.21
Chapter 2.5.2. --- Structure of hormones --- p.22
Chapter 2.5.2.1. --- Structure of growth hormone and prolactin in fish --- p.22
Chapter 2.5.2.2. --- Structure of insulin-like growth factors in fish --- p.26
Chapter 2.5.2.3. --- Structure of Cortisol in fish --- p.27
Chapter 2.5.3. --- Regulation of hormones --- p.28
Chapter 2.5.3.1. --- Regulation of growth hormone in fish --- p.28
Chapter 2.5.3.2. --- Regulation of prolactin in fish --- p.32
Chapter 2.5.3.3. --- Regulation of insulin-like growth factor-I in fish --- p.33
Chapter 2.5.3.4. --- Regulation of Cortisol in fish --- p.33
Chapter 2.5.4. --- Functions of hormones --- p.33
Chapter 2.5.4.1. --- Functions of growth hormone in fish --- p.33
Chapter 2.5.4.2. --- Functions of prolactin in fish --- p.39
Chapter 2.5.4.3. --- Functions of insulin-like growth factor-I in fish --- p.44
Chapter 2.5.4.4. --- Functions of Cortisol in fish --- p.45
Chapter 2.5.4.5. --- "Combined effects of GH, IGF-I, PRL and Cortisol" --- p.49
Chapter 2.6. --- Salinity effects on Na+-K+-ATPase expression --- p.52
Chapter Chapter 3: --- In vitro effect of hormones on branchial Na+-K+- ATPase expression in marine teleost Sparus sarba --- p.58
Chapter 3.1. --- Abstract --- p.58
Chapter 3.2. --- Introduction --- p.60
Chapter 3.3. --- Materials and methods --- p.62
Chapter 3.3.1. --- Overall experimental design --- p.62
Chapter 3.3.2. --- Fish preparation --- p.62
Chapter 3.3.3. --- Tissue sampling --- p.62
Chapter 3.3.4. --- RNA extraction and dot blot analysis --- p.63
Chapter 3.3.5. --- Protein extraction --- p.65
Chapter 3.3.6. --- Protein quantification --- p.65
Chapter 3.3.7. --- Na+-K+-ATPase activity --- p.65
Chapter 3.3.8. --- Protein gel electrophoresis and immunoblotting (Western blotting) --- p.66
Chapter 3.3.9. --- Statistical analysis --- p.67
Chapter 3.4. --- Results --- p.69
Chapter 3.4.1. --- Dot blot analysis of Na+-K+-ATPase mRNA subunits --- p.69
Chapter 3.4.2. --- Analysis of Na+-K+-ATPase protein α-subunit --- p.81
Chapter 3.4.3. --- Analysis of Na+-K+-ATPase activity --- p.87
Chapter 3.5. --- Discussion --- p.92
Chapter 3.5.1. --- Effects of rbGH and rbIGF-I on Na+-K+-ATPase expression --- p.92
Chapter 3.5.2. --- Effects of oPRL on Na+-K+-ATPase expression --- p.102
Chapter 3.5.3 --- Effects of Cortisol on Na+-K+-ATPase expression --- p.104
Chapter 3.6. --- Conclusion --- p.108
Chapter Chapter 4: --- In vivo effect of salinity on branchial Na+-K+-ATPase expression in marine teleost Sparus sarba --- p.109
Chapter 4.1. --- Abstract --- p.109
Chapter 4.2. --- Introduction --- p.110
Chapter 4.3. --- Materials and methods --- p.112
Chapter 4.3.1. --- Overall experimental design --- p.112
Chapter 4.3.2. --- Fish preparation --- p.112
Chapter 4.3.3. --- Tissue sampling --- p.113
Chapter 4.3.4. --- "RNA extraction, dot blot analysis, protein extraction, quantification, Na+-K+-ATPase activity, protein gel electrophoresis and immunoblotting (Western blotting)" --- p.113
Chapter 4.3.5. --- Statistical analysis --- p.114
Chapter 4.4. --- Results --- p.114
Chapter 4.4.1. --- Dot blot analysis of Na+-K+-ATPase mRNA subunits --- p.114
Chapter 4.4.2. --- Analysis of Na+-K+-ATPase protein a-subunit --- p.114
Chapter 4.4.3. --- Analysis of Na+-K+-ATPase activity --- p.115
Chapter 4.5. --- Discussion --- p.120
Chapter 4.6. --- Conclusion --- p.125
Chapter Chapter 5: --- General discussion and conclusion --- p.126
References --- p.130

In the second part of our study, cDNA of AQP-1 and pro-vasotocin were cloned from the silver sea bream. An AQP-1 full clone was isolated from kidney and intestine and it consists of 904 bp with an open reading frame of 774 bp. The deduced amino sequence of sea bream AQP-1 shares highest identity with AQP-1a of gilthead sea bream (97.7%) and AQP-1a of other fish species (83.6% to 95.8%), however, considerably low identity was found between the silver sea bream AQP-1 and AQP-1b of gilthead sea bream (56%). The silver sea bream AQP-1 possesses basic features of a functional aquaporin and AQP-1, which includes two channel-forming asparagine-proline-alanine (NPA) signature motifs, six transmembrane domains, residues of the pore-forming region and a potential mercurial inhibiting site (Cys-178). The water channel was ubiquitously expressed in gills, liver, intestine, rectum, kidney, heart, urinary bladder and blood cells. A partial fragment of pro-vasotocin was isolated from hypothalamus of silver sea bream and consists of 184 bp, including encoding regions for the processing and amidation signal, vasotocin hormone and part of the neurophysin.
Lastly, single doses of cortisol (50 microg/g tissue) or vasotocin (1 microg/g tissue) were administered to seawater-acclimated sea bream with further three-day stabilizing period in seawater followed by an abrupt 6‰ exposure or administered to seawater transfer controls for three days. Cortisol markedly stimulated intestinal expression of AQP-1 in both the seawater transfer control and abrupt 6‰ transfer groups. Vasotocin treatment did not significantly modify AQP-1 expression in all tested organs. Hypothalamic pro-vasotocin expression levels were similar among different treatment groups.
Semi-quantitative RT-PCR analysis was used for studying the effect of salinity and hormones on expression of AQP-1 and pro-vasotocin. In the long-term salinity acclimation experiment, the sea bream were acclimated to six different salinity regimes (0‰, 6‰, 12‰, 33‰, 50‰, 70‰) for four weeks. The abundance of AQP-1 transcript was the highest in intestine of 70‰-acclimated fish among different salinity groups and there was also a statistically significant increase in 12‰-acclimated fish. Branchial AQP-1 expression was significantly upregulated in sea bream acclimated to freshwater. In contrast, the hypothalamic pro-vasotocin expression was significantly downregulated during freshwater acclimation. In addition, the sea bream were also subjected to an abrupt 6%o transfer at different time intervals (2, 6, 12, 72 and 168 hours). RT-PCR analysis revealed there was a transient decrease in branchial AQP-1 expression two hours after abrupt hypo-osmotic exposure and the expression levels subsequently returned to the seawater control levels. The expression levels of hypothalamic pro-vasotocin were not significantly altered by the abrupt exposure treatment.
The present experiments investigated the effects of salinity and hormones on the relative expression of hypothalamic pro-vasotocin, and aquaporin-1 (AQP-1) in intestine, gills and kidney of the silver sea bream Sparus sarba. With the use of immunohistochemical techniques, immunoreactivity of AQP-1 was detected at the basal side of enterocytes and gill chloride cells, and at the apical brush border of kidney tubules whereas AQP-3 was only localized in similar positions in the gills and intestines. AQP-1 was relatively more ubiquitous than AQP-3 and was localized with same cell types as the electrogenic Na+-K+-ATPase in gills and kidney.
The present study had demonstrated the responsiveness of intestinal and branchial AQP-1 expressions of the silver sea bream to environmental salinity perturbations. Further to this, cortisol was observed to upregulate the transcription of AQP-1 in the intestine. Pro-vasotocin expression was altered by long-term salinity adaptation, however, the linkage of this alteration to AQP-1 functioning in different osmoregulatory organs is yet to be elucidated.
Luk, Chun Yin.
Adviser: Norman Y. S. Woo.
Source: Dissertation Abstracts International, Volume: 72-04, Section: B, page: .
Thesis (Ph.D.)--Chinese University of Hong Kong, 2010.
Includes bibliographical references (leaves 200-222).
Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
Electronic reproduction. Ann Arbor, MI : ProQuest Information and Learning Company, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
Abstract also in Chinese.

Ma Wing-chi, Joyce.
Thesis (M.Phil.)--Chinese University of Hong Kong, 2002.
Includes bibliographical references (leaves 131-148).
Abstracts in English and Chinese.
Abstract (in English) --- p.i
Abstract (in Chinese) --- p.iv
Acknowledgement --- p.vi
Contents --- p.viii
Abbreviation --- p.xii
List of Tables --- p.xiii
List of Figures --- p.xv
Chapter 1. --- Introduction --- p.1
Chapter 2. --- Literature review --- p.5
Chapter 2.1 --- Fisheries in Hong Kong --- p.5
Chapter 2.2 --- Flavor of seafood --- p.6
Chapter 2.2.1 --- Lipid-derived volatile aroma compounds --- p.7
Chapter 2.2.2 --- "Alcohols, aldehydes and ketones" --- p.8
Chapter 2.2.3 --- Enzymatic conversion of sulfur- and nitrogen-containing precursors --- p.9
Chapter 2.2.4 --- Thermally generated compounds --- p.9
Chapter 2.2.5 --- Bromophenols --- p.10
Chapter 2.2.5.1 --- General properties of bromophenols --- p.11
Chapter 2.2.5.2 --- Threshold of bromophenols --- p.14
Chapter 2.2.5.3 --- Toxicity of bromophenols --- p.17
Chapter 2.2.5.4 --- Previous studies about bromophenols --- p.19
Chapter 2.2.5.5 --- Bromophenols in aquacultured seafood --- p.20
Chapter 2.2.5.6 --- Possible dietary sources of bromophenols --- p.20
Chapter 2.2.5.7 --- Possibility of increasing bromophenol content in aquacultured fish --- p.23
Chapter 2.3 --- Criteria for selecting experimental fish model --- p.24
Chapter 3. --- Distribution of Bromophenols in selected Hong Kong seafoods --- p.27
Chapter 3.1 --- Introduction --- p.27
Chapter 3.2 --- Materials and methods --- p.28
Chapter 3.2.1 --- Sample collection and preparation --- p.28
Chapter 3.2.2 --- Simultaneous steam distillation-solvent extraction (SDE) --- p.30
Chapter 3.2.3 --- Gas chromatography / mass spectrometry (GC/MS) --- p.30
Chapter 3.2.4 --- Compound identification and quantification --- p.31
Chapter 3.2.5 --- Recoveries --- p.33
Chapter 3.2.6 --- Moisture determination --- p.34
Chapter 3.2.7 --- Statistical analysis --- p.34
Chapter 3.3 --- Results and discussion --- p.34
Chapter 3.3.1 --- Distribution of bromophenols in seafoods --- p.34
Chapter 3.3.1.1 --- Bromophenols in marine fishes --- p.49
Chapter 3.3.1.2 --- Bromophenols in mollusks --- p.49
Chapter 3.3.1.3 --- Bromophenols in crustaceans --- p.50
Chapter 3.3.2 --- Seasonal variations of TBCs --- p.51
Chapter 3.3.3 --- Bromophenols in diet contents --- p.52
Chapter 3.3.4 --- Bromophenol contents of freshwater fish --- p.53
Chapter 3.3.5 --- Relationship between the living habitats and bromophenol contents --- p.56
Chapter 3.3.6 --- Bromophenols as flavor compounds in seafoods --- p.58
Chapter 3.4 --- Conclusion --- p.59
Chapter 4. --- Distribution of Bromophenols in selected Hong Kong seaweeds --- p.61
Chapter 4.1 --- Introduction --- p.61
Chapter 4.2 --- Materials and methods --- p.62
Chapter 4.2.1 --- Sample collection and preparation --- p.62
Chapter 4.2.2 --- Simultaneous steam distillation-solvent extraction (SDE) --- p.63
Chapter 4.2.3 --- Gas chromatography / mass spectrometry (GC/MS) --- p.64
Chapter 4.2.4 --- Compound identification and quantification --- p.65
Chapter 4.2.5 --- Recoveries --- p.66
Chapter 4.2.6 --- Moisture determination --- p.67
Chapter 4.3 --- Results and discussion --- p.67
Chapter 4.3.1 --- Distribution of bromophenols in marine algae --- p.67
Chapter 4.3.2 --- Seasonal variations --- p.76
Chapter 4.3.3 --- Functions of bromophenols in marine algae --- p.79
Chapter 4.3.4 --- Marine algae as sources of bromophenols in marine environment --- p.80
Chapter 4.4 --- Conclusion --- p.81
Chapter 5. --- Enhancement of bromophenol contents in aquacultured fish by the development of bromophenol-rich fish feeds --- p.83
Chapter 5.1 --- Introduction --- p.83
Chapter 5.2 --- Materials and methods --- p.85
Chapter 5.2.1 --- Preparation of fish feeds --- p.85
Chapter 5.2.2 --- Storage conditions of fish feeds --- p.88
Chapter 5.2.3 --- Experimental animals --- p.88
Chapter 5.2.4 --- Solvent and chemicals --- p.90
Chapter 5.2.5 --- Extraction and quantification of bromophenols --- p.90
Chapter 5.2.5.1 --- Simultaneous steam distillation-solvent extraction (SDE) --- p.90
Chapter 5.2.5.2 --- Gas chromatography / mass spectrometry (GC/MS) --- p.91
Chapter 5.2.5.3 --- Compound identification and quantification --- p.92
Chapter 5.2.5.4 --- Recoveries --- p.93
Chapter 5.2.6 --- Moisture determination --- p.94
Chapter 5.2.7 --- Statistical analysis --- p.94
Chapter 5.2.8 --- Sensory test --- p.95
Chapter 5.3 --- Results and discussion --- p.96
Chapter 5.3.1 --- Bromophenol contents in wild-harvested and aquacultured fish --- p.96
Chapter 5.3.2 --- Development of bromophenol-rich fish feed --- p.99
Chapter 5.3.3 --- Effect of feeding the fish with the fish feed developed --- p.105
Chapter 5.3.4 --- Sensory evaluation on the flesh of the fish fed with different fish feeds --- p.121
Chapter 5.3.5 --- Growth of the fish fed with different fish feeds --- p.124
Chapter 5.4 --- Conclusion --- p.126
Chapter 6. --- General conclusion and significance of the study --- p.128
References --- p.131

碩士
國立臺灣海洋大學
生物科技研究所
93
Gonadotropin-releasing hormone (GnRH), which is an essential decapeptide, plays an important role in the control of reproductive function in vertebrates. Three forms of GnRH had been demonstrated in the brains of perciforms including, sea bream GnRH (sbGnRH or GnRH1), chicken GnRHII (cGnRH-II or GnRH2), and salmon GnRH (sGnRH or GnRH3). In this study, complementary as well as genomic DNA, encoding the three forms of GnRH from silver seabream (Sparus sarba), were cloned and sequenced. The genomic structure of GnRH is highly conserved, with 4 exons and 3 introns. The 5’ untranslated region (5’UTR) is encoded by exon I, whereas the signal peptide, GnRH decapeptide, proteolytic cleavage site (Gly-Lys-Arg) and the N-terminus of the GnRH associated peptide (GAP) are encoded by exon II. The exon III encodes the mid region of GAP and exon IV encodes C-terminus of GAP and 3’UTR. Using genome walking strategy, we cloned the promoter regions of sbGnRH (2.6 kb), cGnRH-II (1.9 kb), and sGnRH (3.2 kb) and identified several putative DNA-binding sites of transcription factors such as, Brn-2, progesterone response element (PRE), octamer-binding factor 1 (Oct-1), and GATA binding factor 1 (GATA-1). Furthermore, functional activities of these three promoters were analyzed using a reporter gene in zebrafish (Danio rerio). The expression patterns of sGnRH promoter shown significant specifity of GFP in olfactory bulb-terminal nerve region (OB-TN) and ventral telencephalon-proptic area (VT-POA). However, sbGnRH promoter showed no significant expression and a nonspecific expression was produced by cGnRH-II promoter. These results suggest that cloning of the upstream regulatory region of this gene would provide an opportunity to demonstrate its role in silver seabream and possibility of attempting to ‘knock down’ the expression of this gene to develop a sterile transgenic fish.

碩士
國立臺灣海洋大學
環境生物與漁業科學學系
93
Abstract For understanding the influence of the fish lives along the coastal rocks area and sea Farming-the yellow sea breem（Rhabdosargus sarba）on breed to expect to grow and hatch from eggs by the Tributyltin. The study will research the amount of egg and conceives the rate towards the growth of fish, functions of hatching from eggs, such as rate,genital gland organization variety,G.S.I. value and the H.S.I. value.…..etc., studying the result summary as follows: The yellow sea breem experiment set(2.5μg/l,5μg/l,10μg/l) grows the fish after breeding to expect to was soaked by three stage partition TBTs, in producing the egg for a week start collecting to prognosticate to go to breed the period the last phase, the experiment result is as follows.(1)Breeds to expect each stage to grow the fish individual to equally produce the amount of egg and grow the fish a fertilizied egg quantity with average an Individual, make the data covariance examination result with the ANOVA and the Fisher LSD test,the set of 2.5μg/l of is anti- to its obvious difference(p<0.05), the set of 5μg/l and 10μg/l opposite also all has the obvious difference in the matched control(p<0.05)；the set of 10μg/l after second stage namely stop producing the egg, wasing found by dissection the female and male grows the fish genital gland to all present to atrophy the form.5 μg/l at the third stage after that is stop producing, the egg and genital gland all present to atrophy the form.(2)Breed to expect the matched control and the experimen set of equally conceive the rate, matched control:75.5%(whole periods), the set of 2.5 μg/l:74.9%(whole periods),the set of 5μg/l:68.7%( the first and second stages), the set of 10 μg/l:72.3%(only first stage), statistics the examination with the ANOVA and the Fisher LSD test to all have no obvious difference.(3)Breed to expect the matched control and the experiment set of equally hatch from eggs the rate, matched control:71.5%,(whole periods), the set of 2.5 μg/l:67.3%(whole periods), the set of 5 μg/l:49.6%( the first and second stages), the set of 10 μg/l:32.9%(first stage), depend on to soak the density more high it hatches from eggs the rate more low, statistics the examination with the ANOVA and the Fisher LSD test, at the first stage the set of 10 μg/l contain obvious difference(p<0.05), second stage the set of 5μg/l contain obvious difference(p<0.05), the set of 2.5μg/l then have no obvious difference with the matched control.(4)G.S.I. value first stage ends, statisticsing the examination with the ANOVA and the Fisher LSD test, only the set of 10μg/l of female and male growth the fish contain obvious difference(p<0.05).Another G.S.I. is worth the matched control and experiment a female and male before fish breed expect stage variety the difference was examined by covariance, the growth of male fish in the matched control, the set of 2.5μg/l、5μg/l、10μg/l in front and back the stage all has the obvious difference(p<0.05)；The growth of female fish only the set of 10μg/l in front and back the stage contain obvious difference(p<0.05).(5)The first stage ends and after female and male grow before fish breed expect the stage H.S.I. is worth to change the difference, statisticsing examination through the ANOVA and the Fisher LSD test to all have no obvious difference.(6)After the first stage sample hatch from eggs its fish fry, its matched control and the experiment a value with average S.A.I., was only hads by the covariance examination the set of 10 μg/l contain obvious difference(p<0.05)；Breed the whole period matched control, the set of 2.5 μg/l、5μg/l a covarianceses examine the result to have no obvious difference.(7) the set of 10 μg/l and 5 μg/l adopt to grow the fish genital gland on 24th day after soaking for the second time, find the genital gland all has already atrophied; the set of 2μg/l are breeding the testis of last phase of season to have already presented the degeneration phenomenon, but ovary inside it is thus clear that cumulate ovary cell, show the growing of fish returns to be keeping on the yolk to pile up and producing the egg。

碩士
國立臺灣大學
海洋研究所
99
Silver sea bream (Rhabdosargus sarba) is one of the mariculture sea breams in Taiwan, which grows fast and has high disease resistance. Fishmeal is traditionally the main protein source in mariculture feeds, but a slump in fishmeal production and an increasing demand for formula feeds for mariculture has resulted in the rising costs of fishmeal. It is necessary to replace fishmeal in feeds by using other protein sources. This study was divided into two trials. In the first trial, the control diet contained 60% fishmeal as protein source in the diet and other five experimental diets contained 20-25% soybean meal, 15-20% corn protein, 10% krill meal and 1-5% blood meal and feather meal each. After 42 days of feeding, the experimental group fed diet contained 25% soybean meal, 20% corn protein, 10% krill meal and 1% blood meal and feather meal each had no significant differences in weight gain and feed efficiency rate with the control group. Therefore, the content of fishmeal in diet for silver sea bream could be lowered from 60% to 15%. In the second trial, we used the 60% fishmeal diet in trial 1 as the control diet and test the 15% fishmeal diet in trial 1 once more to check its effect on growth performance of fish. Furthermore, we reduced the content of fishmeal in diet from 15% to 0% by supplying blood meal, feather meal, fish oil and Ca(H2PO4)2. After 42 days of feeding, the experimental group fed non-fishmeal diet showed no significant differences in weight gain and feed efficiency rate with the control group. Therefore, in this study it is suggested that the content of fishmeal in diet for silver sea bream can be reduced to zero with no decrease in growth.

Wong, Sze Ki.
Thesis (M.Phil.)--Chinese University of Hong Kong, 2011.
Includes bibliographical references (leaves 87-106).
Abstracts in English and Chinese.
Acknowledgements --- p.i
Abstract --- p.ii
Abstract in Chinese --- p.iv
Table of contents --- p.V
List of figures --- p.ix
List of abbreviations --- p.X
Chapter Chapter 1. --- General introduction --- p.1
Chapter Chapter 2. --- Literature review --- p.6
Chapter 2.1. --- Pathogenic mechanisms of Vibrio species in fish --- p.7
Chapter 2.1.1. --- Introduction --- p.7
Chapter 2.1.2. --- Adhesion --- p.7
Chapter 2.1.3. --- Invasion --- p.8
Chapter 2.1.4. --- Proliferation --- p.9
Chapter 2.2. --- Vibrio virulence factors --- p.12
Chapter 2.2.1. --- Introduction --- p.12
Chapter 2.2.2. --- Hemolysin --- p.12
Chapter 2.2.3. --- Protease --- p.14
Chapter 2.2.4. --- Siderophore --- p.15
Chapter 2.2.5. --- Lipopolysaccharide --- p.15
Chapter 2.3. --- General apoptotic pathways --- p.17
Chapter 2.3.1. --- Introduction --- p.17
Chapter 2.3.2. --- Extrinsic apoptotic pathway --- p.17
Chapter 2.3.2.1. --- Death receptor signaling apoptosis --- p.17
Chapter 2.3.2.1.1. --- Fas signaling pathway --- p.18
Chapter 2.3.2.1.2. --- TNF-R1 signaling pathway --- p.19
Chapter 2.3.2.1.3. --- TRAIL receptors signaling pathway --- p.20
Chapter 2.3.2.2. --- Growth factor receptor signaling apoptosis --- p.21
Chapter 2.3.3. --- Intrinsic apoptotic pathway --- p.21
Chapter 2.3.3.1. --- Mitochondrial apoptotic pathway --- p.21
Chapter 2.3.3.1.1. --- Cyto c --- p.22
Chapter 2.3.3.1.2. --- Smac/DIABLO --- p.22
Chapter 2.3.3.1.3. --- Omi/HtrA2 --- p.22
Chapter 2.3.3.1.4. --- AIF and endo G --- p.23
Chapter 2.3.3.1.5. --- Bcl-2 family --- p.23
Chapter 2.3.3.1.6. --- Mitochondrial membrane permeabilization (MMP) --- p.23
Chapter 2.3.3.2. --- p53-regulated apoptotic pathway --- p.24
Chapter 2.3.3.3. --- Endoplasmic reticulum (ER) stress-induced apoptotic pathway --- p.25
Chapter 2.4. --- Membrane vesiculation in erythrocytes --- p.26
Chapter 2.4.1. --- Introduction --- p.26
Chapter 2.4.2. --- Induction of vesiculation --- p.26
Chapter 2.4.3. --- Contents of vesicles --- p.28
Chapter 2.4.4. --- Mechanisms involved during vesiculation --- p.29
Chapter 2.4.5. --- Correlation between apoptosis and membrane vesiculation in erythrocytes --- p.31
Chapter 2.4.6. --- Reasons for vesiculation --- p.31
Chapter Chapter 3. --- "Induction of apoptosis by Vibrio alginolyticus thermolabile hemolysin (TLH) in blood cells of silver sea bream, Sparus sarba" --- p.33
Chapter 3.1. --- Abstract --- p.34
Chapter 3.2. --- Introduction --- p.34
Chapter 3.3. --- Materials and methods --- p.36
Chapter 3.3.1. --- Experimental fish --- p.36
Chapter 3.3.2. --- Whole blood preparation --- p.37
Chapter 3.3.3. --- Preparation of V. alginolyticus TLH --- p.37
Chapter 3.3.4. --- "Caspase-3, -8, -9/6 fluorescent assay" --- p.38
Chapter 3.3.5. --- TUNEL assay --- p.39
Chapter 3.3.6. --- Apoptotic DNA ladder assay --- p.40
Chapter 3.3.7. --- Statistical analysis --- p.41
Chapter 3.4. --- Results --- p.42
Chapter 3.4.1. --- "Increase of caspase-3, -8, -9/6 activities" --- p.42
Chapter 3.4.2. --- Detection of DNA fragmentation by TUNEL assay --- p.44
Chapter 3.4.3. --- Detection of DNA fragmentation by apoptotic DNA ladder assay --- p.44
Chapter 3.5. --- Discussion --- p.46
Chapter Chapter 4. --- "Occurrence of membrane vesiculation, apoptosis and post-apoptotic necrosis after exposure to Vibrio alginolyticus thermolabile hemolysin (TLH) in erythrocytes of silver sea bream, Sparus sarba" --- p.51
Chapter 4.1. --- Abstract --- p.52
Chapter 4.2. --- Introduction --- p.52
Chapter 4.3. --- Materials and methods --- p.54
Chapter 4.3.1. --- Experimental fish --- p.54
Chapter 4.3.2. --- Whole blood preparation --- p.54
Chapter 4.3.3. --- Preparation of V. alginolyticus TLH --- p.55
Chapter 4.3.4. --- Light microscopy --- p.55
Chapter 4.3.5. --- Transmission electron microscopy (TEM) --- p.56
Chapter 4.3.6. --- Measurement of membrane vesiculation - acetylcholinesterase (AChE) assay --- p.56
Chapter 4.3.7. --- Measurement of necrosis - hemoglobin colorimetric assay --- p.57
Chapter 4.3.8. --- Apoptotic DNA ladder assay --- p.58
Chapter 4.3.9. --- Flow cytometry --- p.59
Chapter 4.3.10. --- Statistical analysis --- p.59
Chapter 4.4. --- Results --- p.60
Chapter 4.4.1. --- Ultrastructural changes in red blood cells after exposure to TLH --- p.60
Chapter 4.4.2. --- Changes of cell population in size and granularity after exposure of TLH --- p.67
Chapter 4.4.3. --- Effect of TLH dosage on necrosis and DNA fragmentation --- p.72
Chapter 4.4.4. --- "Occurrence of membrane vesiculation, necrosis and DNA fragmentation in cells exposed to TLH" --- p.72
Chapter 4.5. --- Discussion --- p.76
Chapter Chapter 5. --- General conclusions --- p.82
References --- p.87

by Lo Ka-Man.
Thesis (M.Phil.)--Chinese University of Hong Kong, 1997.
Includes bibliographical references (leaves 175-197).
Chapter I --- Title page --- p.i
Chapter II --- Thesis committee --- p.ii
Chapter II --- Acknowledgment --- p.iii-iv
Chapter III --- Abstract --- p.v-vi
Chapter IV --- Table of content --- p.vii-xiv
Chapter V --- List of figures --- p.xv-xviii
Chapter VI --- List of tables --- p.xix-xx
Forewords:
Overall objectives --- p.1
Introduction on the fish used in this research study --- p.2
Chapter Chapter 1: --- Literature Review on Biomarkers of Stress in Teleosts --- p.5
Chapter 1.1 --- Definition of stress --- p.7
Chapter 1.2 --- Classification of stress indicators --- p.8
Chapter 1.2.1 --- Primary stress indicators --- p.8
Chapter 1.2.1.1 --- Molecular stress indicators --- p.8
Chapter 1.2.1.2 --- Hormonal stress indicators --- p.9
Chapter (I) --- Corticosteroid --- p.9
Chapter (II) --- Catecholamines --- p.11
Chapter 1.2.2 --- Secondary stress indicators --- p.12
Chapter 1.2.2.1 --- Metabolic changes --- p.12
Chapter (I) --- Glucose metabolism --- p.13
Chapter (II) --- Lactic acid --- p.14
Chapter 1.2.2.2 --- Osmoregulatory changes --- p.15
Chapter 1.2.2.3 --- Haematological changes --- p.16
Chapter 1.2.2.4 --- Reproductive changes --- p.17
Chapter 1.2.3 --- Tertiary stress indicators --- p.18
Chapter 1.2.3.1 --- Histopathological indicators --- p.18
Chapter 1.2.3.2 --- Ecological indicators --- p.19
Chapter 1.3 --- Recent trends on the study of biomarkers --- p.20
Chapter 1.3.1 --- Use of detoxification enzymes for specific indication of toxic pollutants in aquatic environment --- p.20
Chapter 1.3.1.1 --- Metallothioneins (MTs) --- p.20
Chapter 1.3.1.2 --- Cytochrome P450 monoxygenase (CYP450) --- p.21
Chapter 1.3.2 --- Use of HSP 70 as a biomarker in teleost --- p.22
Chapter 1.4 --- Future perspectives on the study of biomarkers in fish --- p.24
Chapter Chapter 2: --- Literature Review on Heat Shock Proteins (HSPs) --- p.28
Chapter 2.1 --- General Characteristics of HSPs --- p.29
Chapter 2.1.1 --- HSP90 family --- p.30
Chapter 2.1.2 --- HSP70 family --- p.31
Chapter 2.1.3 --- HSP60 family (Chaperonin-60) --- p.32
Chapter 2.1.4 --- Low-molecular weight HSPs (HSP20) --- p.33
Chapter 2.2 --- Structure of HSP70 encoding gene --- p.33
Chapter 2.2.1 --- General characteristics of HSP70-encoding gene --- p.33
Chapter 2.2.2 --- Heat shock transcription factor (HSF) --- p.35
Chapter 2.2.3 --- Heat shock elements (HSE) --- p.35
Chapter 2.3 --- Stress-mediated control of HSP70 transcription --- p.36
Chapter 2.4 --- Characterization of HSP70 expression in teleost --- p.38
Chapter 2.4.1 --- Tissues-specific expression of HSP70 in teleost --- p.39
Chapter 2.4.2 --- Inter-relationship of HSP70 expression with seasonal variation and thermotolerance of teleost --- p.40
Chapter 2.4.3 --- Induction of HSP70 in teleost upon acute thermal stress --- p.41
Chapter 2.4.4 --- Induction of HSP70 in teleost by non-thermal stressors --- p.43
Chapter 2.4.4.1 --- Heavy metal-induced HSP70 expression --- p.43
Chapter 2.4.4.2 --- Handling stress-induced HSP70 expression --- p.43
Chapter Chapter 3: --- "Induction of HSP70 in blood cells of seabream, Sparus sarba subjected to in vivo and in vitro thermal stress" --- p.48
Chapter 3.1 --- Introduction --- p.49
Chapter 3.2 --- Materials and Methods --- p.52
Chapter 3.2.1 --- Overall experimental design --- p.52
Chapter 3.2.2 --- Fish --- p.53
Chapter 3.2.3 --- Blood sampling --- p.53
Chapter 3.2.4 --- Preparation of blood cells --- p.54
Chapter 3.2.5 --- Thermal stress regimes --- p.54
Chapter 3.2.5.1 --- Time couse of HSP70 induction profile in blood cells after in vitro exposure to thermal stress --- p.54
Chapter 3.2.5.2 --- HSP70 synthesis in blood cells taken from fish subjected to in vivo hyper- thermic stress --- p.55
Chapter 3.2.5.3 --- Transcriptional inhibitory effect of actinomycin D on the synthesis of HSP70 in blood cells subjected to in vitro thermal stress --- p.55
Chapter 3.2.6 --- Protein analysis --- p.56
Chapter 3.2.7 --- Gel electrophoresis --- p.57
Chapter 3.2.8 --- Immunoblotting (Western blot analysis) --- p.57
Chapter 3.2.9 --- Autroradiography --- p.58
Chapter 3.3 --- Results --- p.59
Chapter 3.3.1 --- Time course of HSP70 induction profile in blood cells subjected to in vitro thermal treatments --- p.59
Chapter 3.3.1.1 --- Results of immunoblotting from blood cells of fish acclimated to 26°C --- p.59
Chapter 3.3.1.2 --- Results of immunoblotting in blood cells from 18°C-acclimated fish --- p.60
Chapter 3.3.1.3 --- Results of immunoblotting in blood cells from fish acclimated to 20°C --- p.60
Chapter 3.3.1.4 --- 35S-methionine labelling of de novo protein synthesis in blood cells of fish acclimated to 15 and 20°C --- p.61
Chapter 3.3.2 --- Blood cell HSP70 levels in 20°C-acclimated fish subjected to in vivo hyperthermic stress --- p.61
Chapter 3.3.3 --- Transcriptional inhibitory effect of actinomycin D on HSP70 induction in blood cells subjected to in vitro thermal stress --- p.62
Chapter 3.4 --- Discussions --- p.60
Chapter 3.4.1 --- Characteristics of HSP70 induction in blood cells of seabream subjected to in vitro temperature stress --- p.72
Chapter 3.4.1.1 --- Induction profile of HSP70 in blood cells --- p.72
Chapter 3.4.1.2 --- Time course ofHSP70 induction in blood cells --- p.75
Chapter 3.4.1.3 --- Effect of acclimation temperature of fish on the induction of HSP70 --- p.76
Chapter 3.4.2 --- Comparison of HSP70 induction in in vitro and in vivo thermal treatment on blood cells --- p.78
Chapter 3.4.3 --- "Effect of transcriptional inhibitor, actinomycin D, on the de novo synthesis of HSP70" --- p.80
Chapter 3.5 --- Conclusions --- p.70
Chapter Chapter 4: --- "Effects of seasonal variation and transportation stress on level of HSP70, serum glucose and serum Cortisol in seabream, Sparus sarba" --- p.86
Chapter 4.1 --- Introduction --- p.87
Chapter 4.2 --- Materials and methods --- p.90
Chapter 4.2.1 --- Overall experimental design --- p.90
Chapter 4.2.2 --- Fish and blood sampling --- p.91
Chapter 4.2.3 --- Preparation of blood samples --- p.92
Chapter 4.2.4 --- Determination of HSP70 levels in blood cells sampled from seabream upon different seasons --- p.92
Chapter 4.2.5 --- Immunoblotting analysis --- p.92
Chapter 4.2.6 --- Enzyme-linked Immnosorbent Assay (ELISA) --- p.93
Chapter 4.2.7 --- Measurement of serum parameter in seabream --- p.95
Chapter 4.2.7.1 --- Serum glucose --- p.95
Chapter 4.2.7.2 --- Serum Cortisol --- p.96
Chapter 4.3 --- Results --- p.96
Chapter 4.3.1 --- Determination of HSP70 levels in blood cells sampled from seabream in different seasons --- p.96
Chapter 4.3.1.1 --- Immunoblotting analysis --- p.96
Chapter 4.3.1.2 --- Enzyme-linked immunosorbent assay (ELISA) --- p.96
Chapter 4.3.2 --- Serum analysis of seabream sampled from fish farm in different seasons --- p.98
Chapter 4.3.2.1 --- Serum glucose --- p.98
Chapter 4.3.2.2 --- Serum Cortisol --- p.99
Chapter 4.4 --- Discussions --- p.117
Chapter 4.4.1 --- Characterization of HSP70 expression in blood cells of seabream --- p.117
Chapter 4.4.2 --- Dynamicity of HSP70 content and thermo- tolerance of fish in different seasons --- p.118
Chapter 4.4.3 --- Effect of transportation stress on HSP70 induction in blood cells of seabream --- p.121
Chapter 4.4.4 --- Dynamicity of serum glucose level in seabream subjected to seasonal variations --- p.123
Chapter 4.4.5 --- Effect of transportation stress on the serum glucose level of seabream in different seasons --- p.124
Chapter 4.4.6 --- Dynamicity of senam Cortisol level in seabream subjected to seasonal variations --- p.125
Chapter 4.4.7 --- Effect of transportation stress on the serum Cortisol level of seabream in different seasons --- p.126
Chapter 4.4.8 --- "Comments on the use of HSP70, serum Cortisol and serum glucose as biomarkersin environmental supervision" --- p.126
Chapter 4.5 --- Conclusions --- p.129
Chapter Chapter 5: --- "In vitro and in vivo effects of Cortisol, dexamethasone and adrenaline on the induction of HSP70 in seabream, Sparus sarba" --- p.131
Chapter 5.1 --- Introduction --- p.132
Chapter 5.2 --- Materials and methods --- p.133
Chapter 5.2.1 --- Overall experimental design --- p.133
Chapter 5.2.2 --- Acclimation of fish and regimes of treatment --- p.133
Chapter 5.2.3 --- Serum Cortisol and adrenaline analysis --- p.135
Chapter 5.2.4 --- "Protein analysis, gel electrophoresis, immuno- blotting and ELISA analysis" --- p.136
Chapter 5.3 --- Results --- p.137
Chapter 5.3.1 --- "HSP70 level in blood cells treated with Cortisol, dexamethasone and adrenaline in vitro" --- p.137
Chapter 5.3.2 --- "Serum hormones and HSP70 level in tissues of fish injected with Cortisol, adrenaline and dexamethasone invivo" --- p.137
Chapter 5.3.2.1 --- Serum Cortisol and adrenaline level of fish after hormone injections --- p.137
Chapter 5.3.2.2 --- "HSP70 level in blood cells, brain and liver tissue of fish after hormone injections" --- p.138
Chapter (I) --- Level of HSP70 in blood cells of fish after hormone injections --- p.138
Chapter 5.4 --- Discussions --- p.156
Chapter 5.4.1 --- In vitro and in vivo study of the hormonal effect on HSP70 level in blood cells of seabream --- p.156
Chapter 5.4.2 --- Hypothetical mechanism of hormone-receptor mediated HSP70 regulation --- p.158
Chapter 5.4.3 --- In vivo study of the hormonal effect on HSP70 level in blood cells of seabream --- p.160
Chapter 5.4.4 --- In vivo study on the hormonal effect of HSP70 synthesis in liver of seabream --- p.163
Chapter 5.4.5 --- In vivo study on the hormonal effect of HSP70 synthesis in brain tissue of seabream --- p.165
Chapter 5.4.6 --- HSP70 level in different tissues of fish in relation to the induction and sensitivity against stress --- p.166
Chapter 5.5 --- Conclusions --- p.169
Chapter Chapter 6: --- Summary --- p.171
References --- p.175

by Scott P. Kelly.
Thesis (M.Phil.)--Chinese University of Hong Kong, 1994.
Includes bibliographical references (leaves 168-190).
Acknowledgements --- p.iv
Chapter Chapter 1 : --- Introduction --- p.1
Chapter Chapter 2 : --- Review of the literature
Part A : Nutritional requirements of fish and performance assessment
Chapter A2.1 --- Introduction --- p.5
Chapter A2.2 --- Dietary protein requirement --- p.6
Chapter A2.2.1 --- Factors influencing dietary protein requirement
Chapter A2.2.1.1 --- Protein quality
Chapter A2.2.1.2 --- Nonpotein energy
Chapter A2.2.1.3 --- Protein energy to total energy ratio
Chapter A2.2.1.4 --- Fish size/age
Chapter A2.2.1.5 --- Feeding rate
Chapter A2.2.1.6 --- Natural food
Chapter A2.2.1.7 --- Economics
Chapter A2.2.1.8 --- Environmental factors
Chapter A2.3 --- Dietary lipid --- p.14
Chapter A2.3.1 --- Dietary lipid requirement
Chapter A2.4 --- Dietary carbohydrate --- p.16
Chapter A2.4.1 --- Dietary carbohydrate requirement
Chapter A2.5 --- Dietary vitamins
Chapter A2.5.1 --- Dietary vitamin requirements
Chapter A2.6 --- Dietary minerals --- p.19
Chapter A2.6.1 --- Dietary mineral requirements
Chapter A2.7 --- Evaluation criteria --- p.21
Chapter A2.7.1 --- Introduction
Chapter A2.7.2 --- Growth and conversion efficiencies
Chapter A2.8 --- Digestion --- p.22
Chapter A2.9 --- Metabolism in relation to nutritional status --- p.23
Chapter A2.9.1 --- Nitrogen excretion
Chapter A2.9.2 --- Metabolic rate (Oxygen consumption)
Chapter A2.10 --- Biochemical indices of metabolic performance --- p.25
Chapter A2.10.1 --- Tissue composition
Chapter A2.10.1.1 --- Proximate composition and organ indices
Chapter A2.10.1.2 --- Essential amino acid (EAA) profile
Chapter A2.10.2.3 --- Lipid/essential fatty acid (EFA) profile
Chapter A2.10.2.4 --- Tissue RNA/DNA ratio
Chapter A2.11 --- Haematological characteristics --- p.27
Chapter A2.12 --- Enzymes --- p.28
Chapter A2.12.1 --- Enzymes of the intermediate metabolism
Chapter A2.12.2 --- Intestinal enzymes
Chapter A2.13 --- Thyroid hormones --- p.30
Part B : Teleostean salinity adaptation
Chapter B2.1 --- Introduction --- p.31
Chapter B2.2 --- Influence of salinity on growth --- p.32
Chapter B2.3 --- Influence of salinity on weight loss during starvation --- p.37
Chapter B2.4 --- Metabolic rate (oxygen consumption) and salinity adaptation --- p.38
Chapter B2.5 --- Biochemical indices of performance during salinity adaptation --- p.41
Chapter B2.5.1 --- Carcass and tissue composition
Chapter B2.5.2 --- Haematological characteristics
Chapter B2.6 --- Influence of salinity on protein requirements of fish --- p.46
Chapter B2.7 --- Effect of salinity on digestion and digestive enzymes --- p.48
Chapter B2.8 --- Cortisol and osmotic adjustment --- p.50
Chapter B2.9 --- Conclusion --- p.52
Chapter Chapter 3: --- Materials and methods
Chapter 3.1 --- Culture conditions --- p.54
Chapter 3.2 --- Composition of experimental diets --- p.55
Chapter 3.2.1 --- Dietary protein
Chapter 3.2.2 --- Dietary carbohydrate
Chapter 3.2.3 --- Dietary lipid
Chapter 3.2.4 --- Dietary vitamins
Chapter 3.2.5 --- Dietary minerals
Chapter 3.2.6 --- Chromic oxide
Chapter 3.2.7 --- Binder
Chapter 3.3 --- Proximate analysis of experimental diets --- p.61
Chapter 3.3.1 --- Moisture
Chapter 3.3.2 --- Lipid
Chapter 3.3.3 --- Protein
Chapter 3.3.4 --- Ash
Chapter 3.3.5 --- Nitrogen free extractives
Chapter 3.3.6 --- Metabolizable energy
Chapter 3.4 --- Dietary evaluation criteria --- p.65
Chapter 3.4.1 --- Growth rate
Chapter 3.4.2 --- Protein efficiency ratio
Chapter 3.4.3 --- Food conversion ratio
Chapter 3.4.4 --- Feed conversion efficiency
Chapter 3.5 --- Digestibility --- p.66
Chapter 3.6 --- Metabolism --- p.67
Chapter 3.6.1 --- Ammonia excretion
Chapter 3.6.2 --- Oxygen consumption
Chapter 3.7 --- Biochemical analysis
Chapter 3.7.1 --- Organ indices
Chapter 3.7.2 --- Serum metabolites
Chapter 3.7.2.1 --- Collection of serum
Chapter 3.7.2.2 --- Serum ions
Chapter 3.7.2.3 --- Serum protein
Chapter 3.7.2.4 --- Serum α-amino acids
Chapter 3.7.2.5 --- Serum ammonia
Chapter 3.7.2.6 --- Serum glucose
Chapter 3.7.2.7 --- Serum lipids
Chapter 3.7.2.8 --- Serum hormones
Chapter 3.7.3 --- Proximate analysis of white muscle --- p.74
Chapter 3.7.4 --- Analysis of hepatic tissue --- p.75
Chapter 3.7.4.1 --- Proximate analysis
Chapter 3.7.4.2 --- Liver glycogen
Chapter 3.7.4.3 --- Hepatic enzymes
Chapter 3.7.4.3. --- a Glucose-6-phosphatase
Chapter 3.7.4.3. --- b Glucose-6-phosphate dehydrogenase
Chapter 3.7.4.3. --- C Hexokinase
Chapter 3.7.5 --- Intestinal enzymes --- p.80
Chapter 3.7.5.1 --- γ-Glutamyltranspeptidase
Chapter 3.7.5.2 --- Trypsin
Chapter 3.7.5.3 --- α-Amylase
Chapter 3.7.6 --- Statistical analysis --- p.83
Chapter Chapter 4: --- Results
Chapter 4.1 --- Growth and conversion efficiencies --- p.84
Chapter 4.2 --- Metabolism --- p.90
Chapter 4.2.1 --- Ammonia excretion
Chapter 4.2.2 --- Oxygen consumption
Chapter 4.3 --- Biochemical analysis --- p.93
Chapter 4.3.1 --- Organ indices --- p.93
Chapter 4.3.2 --- Serum metabolites --- p.98
Chapter 4.3.2.1 --- Serum ions
Chapter 4.3.2.2 --- "Serum protein, α-amino acids and ammonia"
Chapter 4.3.2.3 --- Serum glucose
Chapter 4.3.2.4 --- Serum lipids
Chapter 4.3.2.5 --- Serum hormones
Chapter 4.3.3 --- Proximate composition of white muscle --- p.113
Chapter 4.3.4 --- Composition of hepatic tissue --- p.117
Chapter 4.3.4.1 --- Proximate composition and liver glycogen
Chapter 4.3.4.2 --- Hepatic enzymes
Chapter 4.3.5 --- Total digestibility --- p.126
Chapter 4.3.6 --- Intestinal enzymes --- p.126
Chapter Chapter 5: --- Discussion
Chapter 5.1 --- Growth and conversion efficiencies --- p.132
Chapter 5.2 --- Metabolism --- p.135
Chapter 5.2.1 --- Ammonia excretion
Chapter 5.2.2 --- Oxygen consumption
Chapter 5.3 --- Organ indices --- p.137
Chapter 5.4 --- Serum metabolites --- p.140
Chapter 5.4.1 --- Serum ions
Chapter 5.4.2 --- "Serum protein, α-amino acids and ammonia"
Chapter 5.4.3 --- Serum glucose
Chapter 5.4.4 --- Serum lipids
Chapter 5.4.5 --- Serum hormones
Chapter 5.5 --- Proximate composition of white muscle --- p.150
Chapter 5.6 --- Proximate composition of liver --- p.152
Chapter 5.7 --- Hepatic enzymes
Chapter 5.7.1 --- Glucose-6-phosphate dehydrogenase (G6P-DH)
Chapter 5.7.2 --- Hexokinase
Chapter 5.7.3 --- Glucose-6-phosphatase (G-6-Pase)
Chapter 5.8 --- Total digestibility --- p.159
Chapter 5.9 --- Intestinal enzymes
Chapter 5.9.1 --- Trypsin
Chapter 5.9.2 --- γ-Glutamyltranspeptidase (γ-GT)
Chapter 5.9.3 --- α-Amylase
Chapter Chapter 6: --- Conclusion --- p.164
References --- p.168

碩士
國立澎湖科技大學
水產養殖系水產資源與養殖碩士班
105
Marine fish larvae often consume rotifers as their initial food organisms when they begin feeding, but many species are not able to feed until they find rotifers of a suitable size. The salinity for optimal rotifer proliferation is 14 psu (Koiso and Hino,2001), which is considerably different from the seawater conditions at the time of larval fish production. Therefore, the aim of the present study was to determine the most suitable experimental salinity for rotifer culture, and obtain smaller rotifers using different methods; and to observe the effects of feeding with the smaller rotifers on the growth of fish larvae. Rotifers were initially cultured in seawater with salinities of 32, 26, 20, and 14 psu, and were then stirred vigorously, after stirred with blender, the shakedown parthenogenesis eggs were collected to hatch and then to collect the newly hatched rotifer, or using plankton net filtering can be applied to collect the small-size rotifer larvae which is below 150 μm. Unscreened rotifers were used as the control group. The rotifers were fed to larvae of two marine fish (Rhabdosargus sarba and Epinephelus coioides) of different sizes up to 240 h after hatching. Fish larvae (n = 30) were sampled randomly every 3–6 h, fixed in 3% formalin, and measured. The results indicated the proliferation rate, carrying egg rate, and tolerance to environmental changes of rotifers cultivated at a salinity of 20 psu were higher than those of other salinity groups. When the carrying egg rate was higher than 80%, it was suggested that the rotifer eggs should be collected after vigorous shaking 6 h after hatching; and when the carrying egg rate was low, fewer small-sized rotifers should be collected using a filtering method more frequently, and the duration out of water should be decreased where possible. There was no significant difference with respect to the growth of R. sarba larvae between the unscreened and small-sized rotifer feeding groups, whereas the growth of E. coioides larvae fed small-sized rotifers was significantly higher than that of the unscreened group from 168 h after hatching.

Wong Kwok-Shing.
Thesis (M.Phil.)--Chinese University of Hong Kong, 2001.
Includes bibliographical references (leaves 89-107).
Abstracts in English and Chinese.
Title --- p.i
Abstract (English) --- p.ii
Abstract (Chinese) --- p.v
Content --- p.vii
Acknowledgement --- p.x
Abbreviation --- p.xii
Lists of tables and figures --- p.xiii
Chapter Chapter 1 --- General introduction --- p.1
Chapter Chapter 2 --- "Effects of salinity on the cardiovascular responses and dipsogenic behaviors and silver seabream, Sparus sarba."
Chapter 2.1 --- Literature review
Chapter 2.1.1 --- Teleost euryhalinity --- p.5
Chapter 2.1.2 --- Salinity and blood respiratory properties --- p.7
Chapter 2.1.3 --- Salinity and blood volume --- p.8
Chapter 2.1.4 --- Salinity and blood pressure --- p.10
Chapter 2.1.5 --- Intestine physiology --- p.12
Chapter 2.1.6 --- Summary --- p.14
Chapter 2.2 --- Materials and methods
Chapter 2.2.1 --- Experimental animals --- p.19
Chapter 2.2.2 --- Salinity adaptation --- p.19
Chapter 2.2.3 --- Drinking rate measurement --- p.19
Chapter 2.2.4 --- Respiratory characteristics --- p.20
Chapter 2.2.5 --- Blood volume measurement --- p.21
Chapter 2.2.6 --- Blood pressure experiment --- p.23
Chapter 2.2.7 --- Statistical analysis --- p.23
Chapter 2.3 --- Results
Chapter 2.3.1 --- Drinking rate --- p.25
Chapter 2.3.2 --- Oxygen dissociation curves --- p.27
Chapter 2.3.3 --- Blood volume --- p.29
Chapter 2.3.4 --- Blood pressure --- p.31
Chapter 2.4 --- Discussion
Chapter 2.4.1 --- Drinking rate --- p.36
Chapter 2.4.2 --- Oxygen dissociation curves --- p.37
Chapter 2.4.3 --- Blood volume --- p.38
Chapter 2.4.4 --- Blood pressure --- p.40
Chapter Chapter 3 --- "Manipulation of renin-angiotensin system in relation to the cardiovascular responses and dipsogenic behaviors of silver seabream, Sparus sarba."
Chapter 3.1 --- Literature review
Chapter 3.1.1 --- Renin angiotensin system (RAS) --- p.41
Chapter 3.1.2 --- RAS and blood pressure --- p.47
Chapter 3.1.3 --- RAS and drinking --- p.53
Chapter 3.1.4 --- RAS and Cortisol --- p.55
Chapter 3.1.5 --- RAS and kidney --- p.58
Chapter 3.1.6 --- Summary --- p.58
Chapter 3.2 --- Materials and methods
Chapter 3.2.1 --- Experimental animals --- p.61
Chapter 3.2.2 --- Salinity adaptation --- p.61
Chapter 3.2.3 --- Drinking rate measurement --- p.61
Chapter 3.2.4 --- Determination of angiotensin converting enzyme (ACE) activity --- p.61
Chapter 3.2.5 --- Blood pressure experiment --- p.62
Chapter 3.2.6 --- Statistical analysis --- p.63
Chapter 3.3 --- Results
Chapter 3.3.1 --- Drinking rate --- p.64
Chapter 3.3.2 --- ACE activity --- p.69
Chapter 3.3.3 --- Blood pressure --- p.71
Chapter 3.4 --- Discussion
Chapter 3.4.1 --- Drinking rate --- p.77
Chapter 3.4.2 --- ACE activity --- p.81
Chapter 3.4.3 --- Blood pressure --- p.83
Chapter Chapter 4 --- General conclusion --- p.86
Reference --- p.89

碩士
國立臺灣海洋大學
水產養殖學系
94
Abstract P450 aromatase (P450arom, CYP19), a cyp19 gene product, is an important enzyme in the steroidogenic pathway and belongs to the cytochrome P450 superfamily. Aromatase is responsible for the conversion of androgen into estrogen and is found throughout the vertebrate phylum. Most teleosts have two isoforms of the cyp19 gene, ovary type and brain type, termed cyp19a and cyp19b. In the present study, the reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA end (RACE) were applied to clone the silver sea bream (Rhabdosargus sabar) cyp19a and cyp19b genes. Full length cDNA of silver sea bream cyp19a is 1842 bp, containing 50 bp in 5’-untranslation region, 232 bp in 3’-untranslation region and 1560 bp in coding region which can be deduced 519 amino acids. The full length cDNA of silver sea bream cyp19b is 1916 bp, containing 276 bp in 5’-untranslation region, 137 bp in 3’-untranslation region and 1503 bp in coding region which can be deduced 500 amino acids. The 3D structure of CYP19a is comprised of membrane-targeting segment, transmembrane helix, I-helix, steroid binding helix (Ozol’s peptide region) and heme-binding helix. Comparison of the deduced amino acid sequence of silver sea bream cyp19a to that of the cyp19b revealed only a 59% similarity. The real-time quantitative PCR was performed to study tissue-specific expression of both cyp19a and cyp19b gene in adult silver sea bream. Cyp19a was mainly expressed in silver sea bream gonads, both ovary and testis, but was also found at low levels in the kidney. In contrast, cyp19b was expressed at higher levels in brain of silver sea bream than that of female silver sea bream. Zebrafish and tilapia during the period of gonadal differentiation were treated with the non-steroidal aromatase inhibitor (formestane) causing masculinization. The gonad development, oocyte growth and spermatogenesis is suppressed with aromatase inhibitor in a dose-dependent manner.