Molt and reproduction enhancement together with hemolymph ecdysteroid elevation under eyestalk ablation in the female fiddler crab,Uca triangularis(Brachyura: Decapoda)

We present the results of eyestalk extirpation experiments performed on the fiddler crab, Uca triangularis at seasons of molting and reproduction, with a view to have a better understanding of the regulatory mechanisms of these two highly energy-demanding processes. Bilateral eyestalk ablation resulted in precocious acceleration of both molting and reproduction, irrespective of the season at which each experiment was conducted. The rate of accelerated ovarian growth, however, was maximum if the eyestalk ablation was conducted during August–January, the breeding season in the wild, or in February–May(molting-reproductive season), wherein a section of the wild population would be engaged in molting and another section in breeding. The highest degree of precocious molt acceleration, on the other hand, was obtained during June–July when the population was primarily engaged in molting, but with no reproductive activity. The precocious oocyte maturation(due to de-eyestalking) was minimal in June–July. Significantly, the eyestalk ablation also resulted in a dramatic increase in the hemolymph ecdysteroid titer, revealing that a high ecdysteroid titer would have no restraining influence on vitellogenesis. No spawning was, however, observed among de-eyestalked females, even though their final oocyte size surpassed the size of the normal mature oocytes, implying that spawning is not exclusively under the control of eyestalk hormones. A comparative study performed on the vitelline components of the experimentals and the controls revealed that the precociously incorporated yolk under eyestalk ablation was biochemically impoverished. These results indicate that throughout the annual cycle, both the somatic and the reproductive growth of U. triangularis are under the influence of inhibitory principles from the eyestalks. It is also revealing that mere deprival of the inhibitory principles does not culminate into successful vitellogenesis. Arguably, the inhibitory influence from the eyestalks could be a prerequisite for normal healthy maturation of the oocytes and spawning.     

Seasonal Gametogenesis of Host Sea Anemone (Entacmaea quadricolor) InhabitingHong Kong Waters

Studying gonadal development of annual cycle can reveal the process of gametogenesis and reproductive period, and evaluate fertility and source utilization of a species. Host sea anemones are conspicuous members of tropical and subtropical reef ecosystems, but little is known about its biology including reproductive seasonality. Here we reported a one-year study on the gametogenesis and reproduction of host sea anemone(Entacmaea quadricolor) inhabiting Hong Kong waters. E. quadricolor tissues were sampled in 12 occasions from 5 m and 15 m depths of water, respectively. Histological sectioning of the tissues showed that E. quadricolor was dioecious, and populational ratio of female to male was 1:1.6. The gonadal development was asynchronous within an annual cycle, which included proliferating, growing, maturing, spawning, and resting stages. The spawning occurred between August and October when surface seawater temperature reached the annual maximum(28℃), suggesting that temperature is an important factor modulating the gonadal development and mature of E. quadricolor.     

The effects of temperature and substrate on ontogenetic behavior of bastard halibut, Paralichthys olivaceus （T.et S.）

The effects of temperature and substrate on ontogenetic patterns and settlement behavior of bastard halibut, Paralichthys olivaceus （T. et S.） were studied in a series of laboratory experiments. Analysis on stage-based data revealed that except for settlement, and compared to the substrate, the ambient temperature dominantly controlled the hatch success and subsequent development of the species. The oosperm optimum survival rate of 60% during hatching occurred at 20℃, and survival rate of 29%, at 18℃ for larval rearing. The survival rate of larvae reared under variable temperature of 3.5-4.5℃ was higher than that of those reared under stable temperature. The maximum survival rate of 29%, corresponded to variation at temperature of 4℃. With increase in age newly settled juveniles preferred to lie on the fine substrate and gradually moved on the coarse substrate. GLM （general linear model） analysis showed that the combination of temperature and substrate had no significant impact on the survival of settling larvae, but definitely affected the duration of settlement.     

LABORATORY OBSERVATION ON SPAWNING,FECUNDITY AND LARVAL DEVELOPMENT OF AMPHIOXUS (BRANCHIOSTOMA BELCHERI TSINGTAUNESE)

Although amphioxus is widespread in temperate and tropical seas,its population is diminishing be-cause of environmental pollution.To keep the population of this evolutionarily important animal fromdiminishing,study on its reproduction and development is necessary.The main findings in this study onthe spawning and fecundity of the amphioxus reared in laboratory and its larval development are as fol-lows.1.Water temperature markedly affected the spawning.It spawned only when water temperature reached 21℃.2.Spawning of the amphioxus in laboratory was markedly extended.Initially,the amphioxus spawned at about 7:00 PM,but spawning time was postponed as spawning days went on.3.The number of eggs produced by a female ranged from 1400 to 12800,average of 5800.This al-so represents the fecundity of the amphioxus because it shedded all eggs within the ovary at a time.4.During the first few months of life of the amphioxus,its growth rate changed seasonally.Thegrowth rate in summer and fall was greater than that in     

Characterization and differential expression of three GnRH forms during reproductive development in cultured turbot Schophthalmus maximus

Turbots( Schophthalmus maximus), one of the most important economic marine flatfish species, fail to undergo final spawning and spermiation naturally under artificial farming conditions. In vertebrates, reproduction is regulated by the brain-pituitary-gonadal axis(BPG-axis), and gonadotropin releasing hormone(GnRH) is one of its key components. Therefore, to better understand the physiology of reproduction in the turbot, three of the genes encoding GnRH subtypes— sbGnRH, c GnRH-II and sGnRH —were cloned and sequenced by isolating the cDNA sequences. The localizations and patterns of expression of their mRNAs were also evaluated during seasonal gonadal development. All three mRNAs were expressed abundantly in the brain; sbGnRH and sGnRH mRNAs were also detected in the gonads and pituitary gland, and sbGnRH expression was much higher than that of sGnRH, indicating the critical role of sbGnRH in regulating the BPG-axis. Moreover, the brain expression patterns of sbGnRH and sGnRH mRNAs showed an increased trend during gonadal development, peaking in mature stages. This indicated the direct regulation of gonadal development by the GnRH system. In addition, c GnRH-II mRNA expression showed no significant variations, suggesting that c GnRH-II is not critically involved in the control of reproduction. Further, the mRNA abundances of the three GnRH forms in the breeding season were significantly higher than those in immature and post-breeding stages in all analyzed brain areas. Therefore, we propose that sbGnRH is the most important hormone for the regulation of reproduction in turbot via the BPG-axis. These results will help in better understanding the reproductive endocrine mechanisms of turbots and lay the groundwork for additional studies aimed at comparing the reproductive physiology of wild individuals with those raised under artificial conditions.     

Effect of diurnal temperature difference on lipid accumulation and development in Calanus sinicus(Copepoda:Calanoida)

Calanus sinicus,the dominant copepod in the Yellow Sea,develops a large oil sac in late spring to prepare for over-summering in the Yellow Sea Cold Water Mass(YSCWM).The lipid accumulation mechanism for the initiation of over-summering is unknown.Here,we cultured C3 copepodites at four constant temperatures(10,13,16,and 19℃) and at three temperature regimes that mimicked the temperature variations experienced during diurnal vertical migration(10-13℃,10-16℃,and 10-19℃) for 18 days to explore the effects of temperature differences on copepod development and lipid accumulation.C.sinicus stored more lipid at low than at high temperatures.A diurnal temperature difference(10-16℃ and 10-19℃)promoted greater lipid accumulation(1.9-2.1 times) than a constant temperature of either 16℃ or 19℃,by reducing the energy cost at colder temperatures and lengthening copepodite development.Thereafter,the lipid reserve supported gonad development after final molting.Only one male developed in these experiments.This highly female-skewed sex ratio may have been the result of the monotonous microalgae diet fed to the copepodites.This study provides the first evidence that diurnal temperature differences may promote lipid accumulation in C.sinicus,and provides a foundation for future investigations into the mechanisms involved in over-summering in the YSCWM.     

Breeding and mass scale rearing of clownfish Amphiprion percula: feeding and rearing in brackishwater

Breeding and mass scale larval rearing of clownfish Amphiprion percula is very limited in brackishwater. We designed an indoor program of A. percula culture in brackishwater with a salinity of 24±1, during which the impacts of feed type, water temperature, and light intensity, on the efficiency of its reproduction, were revealed. The fish were accommodated along with sea anemones in fibre glass tanks to determine the influence of brooder diet on breeding efficiency. Higher reproductive efficiency [number of eggs laid (276 22.3 eggs)] was observed when fish were fed live Acetes sp. rather than clam (204 16.4 eggs), trash fish (155 12 eggs) and formulated feed (110 10 eggs). The spawning rate was increased during September and October (water temperature, 28.74 0.55°C) on average of 2.4 spawning per month; and low spawning rate was in January (water temperature, 24.55 0.45°C) on average of 1 spawning per month. Among three light intensities (100, 500, and 900 lx) set to evaluate larval survival rate, larvae showed the highest survival rate (65.5%) at 900 lx. The breeding method specifically in brackishwater developed in the present study is a new approach, will help the people from the regions of estuary and backwater to enhance their livelihood and it will lead to reduce the exploitation from the wild habitat.     

Early development of Costaria costata (C. Agardh) Saunders and cultivation trials

Costaria costata(C.Agardh) Saunders is one of common kelps distributed in many coastal areas worldwide;however,in China,no reports have been made on cultivation of the genus.To investigate potential cultivation of the species in the northern part of China,trials on isolation and preservation of the gametophytes were conducted using C.costata from Korea;growth and development of the gametophytes were observed.We showed that at 10±1°C,60 μmol m-2s-1 and 12:12 h(L:D),freshly released zoospores settled down wit...     

Effects of temperature and salinity on the development of embryos and larvae of the veined rapa whelk Rapana venosa （Valenciennes, 1846）

The major population of the veined rapa whelk Rapana venosa(Valenciennes, 1846), which is an important fishery resource, is facing a large decline in China. We studied the effects of incubation temperature(16–34℃ at salinity 30) and salinity(5–45 at 25℃) on the incubation period and subsequent larval development. In the temperature experiment, the shortest incubation period was 12 days at 34℃, the lower temperature limit was 16℃, the longest mean shell length(1 193±17 μm) occurred at 25℃ and the highest survival rate 72.28%±5.62% was observed at 28℃. In the salinity experiment, the shortest incubation period was 15 days at 25. The salinity tolerance range was 15–40, the longest mean shell length(855±9 μm) and the highest survival rate 72.93%±4.85% were both observed at 35. This study demonstrated that, during the egg–mass stage, temperature and salinity regimes infl uence later growth and survival of larvae. These observations deepen our understanding of the ecology and conservation of natural populations of Rapana venosa.     

Effects of temperature and irradiance on early development of Chondrus ocellatus Holm(Gigartinaceae, Rhodophyta)

Chondrus is a type of commercially produced red seaweed that widely used for food and carrageen extraction. Although the natural life history of the alga had been well understood, the factors influencing development of the tetraspore and carpospore remain poorly understood. In the perspective of seedling resources, the regulation of early development is crucial for the seedling nursing; therefore, it is necessary to understand the physiological influences during its early development. In this study, we studied the effects of temperature and irradiance on the early development of Chondrus ocellatus Holm under laboratory conditions. The released tetraspores and carpospores were cultivated at different temperatures (10–28°C) and irradiances (10, 60 μmol photons m-2s-1) with a photoperiod of 12L:12D. The results indicate that both tetraspores and carpospores are tolerant to temperatures of 10–25°C, and have the highest relative growth rate at 20°C. Irradiance variances influenced the growth of the discoid crusts, and the influence was more significant with increasing temperature; 60 μmol photons m-2s-1 was more suitable than 10 μmol photons m-2s-1. The optimum temperature and irradiance for the development of seedlings was 20°C and 60 μmol photons m-2s-1, respectively.     

Effect of temperature on gene expression in the pearl oyster Pinctada fucata

In this study, we examined the effect of elevated temperature on the expression patterns of genes, i.e., nacrein, irr, n16, n19, and hsp70 in the pearl oyster Pinctada fucata. The experiment was carried out at 4 temperatures, i.e., 20℃(ambient, control), 24, 28℃, and 32℃. The expression levels of target genes in P. fucata were assayed at 0, 6, 24, 48, and 96 h via real-time polymerase chain reaction. Results showed that the expression levels of nacrein and irr had no significant variations among different time points below 28℃, but significantly increased over time at 32℃. The expression levels of n16 and n19 did not change markedly at 20℃. The former increased significantly at 6 h and 24 h while the latter substantially decreased during 6–96 h at 24, 28 and 32℃. Among different temperatures, the level of n16 was significantly lower at 20℃ than at other temperatures during 6–96 h, and the level of n19 significantly varied among different temperatures at 48 h and 96 h. The expression level of hsp70 was significantly higher at 32℃ than at 20, 24 and 28℃ at 24 h. These results demonstrated that elevated temperature impacted the physiological processes of P. fucata and potentially influenced its adaptability to thermal stress.     

Seasonal variation of biochemical components in clam (<Emphasis Type="Italic">Saxidomus purpuratus</Emphasis> Sowerby 1852) in relation to its reproductive cycle and the environmental condition of Sanggou Bay,China

Seasonal variation of biochemical components in clam (Saxidomus purpuratus Sowerby 1852) was investigated from March 2012 to February 2013 in relation to environmental condition of Sanggou Bay and the reproductive cycle of clam. According to the histological analysis, the reproductive cycle of S. purpuratus includes two distinctive phases: a total spent and inactive stage from November to January, and a gametogenesis stage, including ripeness and spawning, during the rest of the year. Gametes were generated at a low temperature (2.1°C) in February. Spawning took place once a year from June to October. The massive spawning occurred in August when the highest water temperature and chlorophyll a level could be observed. The key biochemical components (glycogen, protein and lipid) in five tissues (gonad, foot, mantle, siphon and adductor muscle) were analyzed. The glycogen content was high before gametogenesis, and decreased significantly during the gonad development in the gonad, mantle and foot of both females and males, suggesting that glycogen was an important energy source for gonad development. The protein and lipid contents increased in the ovary during the gonad development, demonstrating that they are the major organic components of oocytes. The lipid and protein contents decreased in the testis, implying that they can provide energy and material for spermatogenesis. The results also showed that protein stored in the mantle and foot could support the reproduction after the glycogen was depleted.     

Dietary vitamin A,ascorbic acid and α-tocopherol affect the gonad development and reproductive performance of starry flounder Platichthys stellatus broodstock

The present trial was conducted with starry fl ounder Platichthys stellatus broodstock to evaluate the effects of dietary vitamin A,ascorbic acid,and α-tocopherol on the gonadal development and reproductive performance.8 000 IU/kg diet vitamin A(VA group),500 mg/kg diet ascorbic acid(Vcpp group),or 250 mg/kg diet α-tocopherol(α-TA group)was added into basal diet to create 3 vitamin experimental diets,respectively.Each diet was fed to 450 starry fl ounder broodstock for 104 days.Samples were collected weekly.The gonadosomatic index(GSI)of 3 vitamin experimental groups fi rst decreased and then increased.Maximum GSI of Vcpp group was higher than that of α-TA group but lower than that of VA group.The spawning periods of 3 vitamin experimental groups lasted 49,56,and 45 days,respectively.No mature eggs were observed in the control group during the trial.The absolute fecundity(AF)and relative fecundity(RF)of α-TA group was higher than that of Vcpp group but lower than that of VA group.The results suggest that different vitamins play different roles in the fi sh reproductive process.Vitamin A stimulated the maturation of the ovary,ascorbic acid prolonged the spawning period,and α-tocopherol affected the development of the eggs.     

Effect of water temperature on digestive enzyme activity and gut mass in sea cucumber Apostichopus japonicus （Selenka）, with special reference to aestivation

The effect of water temperature on gut mass and digestive enzyme activity in sea cucumber Apostichopus japonicus, including relative gut mass (RGM), amylase, lipase, pepsin and trypsin activities were studied at temperatures of 7, 14, 21, and 28°C over a period of 40 days. Results show that RGM significantly decreased after 40 days at 21°C and markedly decreased over the whole experiment period at 28°C; however, no significant effect of duration was observed at 7 or 14°C. At 14°C, trypsin activity significantly decreased over 10 and 20 days, then increased; amylase and trypsin activity significantly decreased after 40 days at 28°C. However, no significant effect of duration was found on amylase, pepsin or trypsin activities in the other temperature treatment groups. At 28°C, lipase activity peaked in 20 days and then markedly decreased to a minimum at the end of the experiment. On the other hand, pepsin activity at 28°C continuously increased over the whole experimental period. Principle component analysis showed that sea cucumbers on day 40 in the 21°C group and in the previous 20 days in the 28°C group were in the prophase of aestivation. At 28°C, sea cucumbers aestivated at 30–40 days after the start of the experiment. It is concluded that the effect of temperature on the digestion of A. japonicus is comparatively weak within a specific range of water temperatures and aestivation behavior is accompanied by significant changes in RGM and digestive enzyme activities.     

Pharmacokinetics and tissue behavior of enrofloxacin and its metabolite ciprofloxacin in turbot Scophthalmus maximus at two water temperatures

Turbot Scophthalmus maximus, an important aquaculture species in China, currently suffers from epizootic diseases because of high density aquaculture. Enrofloxacin has been used to treat various systemic bacterial fish infections. However, studies concerning the pharmacokinetics of enrofloxacin in turbot are limited. In this study, the pharmacokinetics of enrofloxacin and its metabolite ciprofloxacin, were investigated in the turbot following intravenous and oral administration at 10 mg enrofloxacin/kg body weight, at 16°C and 10°C water temperatures. The concentrations of enrofloxacin and ciprofloxacin in the main tissues (plasma, muscle, liver and kidney) were detected by HPLC. The results show that the plasma concentration-time data for enrofloxacin were best described as a two-compartment open model after intravenous and oral administration. Three pharmacokinetic equations were established between the concentrations and temperatures. The kinetic profile of enrofloxacin was temperature dependent. The absorption half-life of enrofloxacin was 1.99 h and 2.17 h after oral administration, whereas the elimination half-life of the drug was 98.63 h and 136.59 h at 16°C and 10°C, respectively. The peak concentration of enrofloxacin in plasma and tissues was higher at 16°C than that at 10°C, and the peak plasma concentration time in the liver was the shortest at both temperatures among those of other tissues. The plasma C max /MIC ratio varied between 11.08 and 5 540.00 at 16°C; and between 7.92 and 3 960.00 at 10°C. The AUC/MIC ratio was 467.82-280 690.00 at 16°C, and 359.48-215 690.00 at 10°C. These ratios indicate that it is possible to obtain therapeutic efficacy. Very low levels of ciprofloxacin were detected. The AUC ratios of ciprofloxacin and enrofloxacin in plasma suggest that plasma ciprofloxacin might play a minor role in enrofloxacin treatment for turbot.     

Effects of temperature and irradiance on early development of Chondrus ocellatus Holm (Gigartinaceae, Rhodophyta)

Chondrus is a type of commercially produced red seaweed that widely used for food and carrageen extraction. Although the natural life history of the alga had been well understood, the factors influencing development of the tetraspore and carpospore remain poorly understood. In the perspective of seedling resources, the regulation of early development is crucial for the seedling nursing; therefore, it is necessary to understand the physiological influences during its early development. In this study, we studied the effects of temperature and irradiance on the early development of Chondrus ocellatus Holm under laboratory conditions. The released tetraspores and carpospores were cultivated at different temperatures (10–28°C) and irradiances (10, 60 μmol photons m⁻²s⁻¹) with a photoperiod of 12L:12D. The results indicate that both tetraspores and carpospores are tolerant to temperatures of 10–25°C, and have the highest relative growth rate at 20°C. Irradiance variances influenced the growth of the discoid crusts, and the influence was more significant with increasing temperature; 60 μmol photons m⁻²s⁻¹ was more suitable than 10 μmol photons m⁻²s⁻¹. The optimum temperature and irradiance for the development of seedlings was 20°C and 60 μmol photons m⁻²s⁻¹, respectively.     

Diurnal and seasonal variation of the elevation gradient of air temperature in the northern flank of the western Qinling Mountain range,China

The typically sparse or lacking distribution of meteorological stations in mountainous areas inadequately resolves temperature elevation variability. This study presented the diurnal and seasonal variations of the elevation gradient of air temperature in the northern flank of the western Qinling Mountain range,which has not been thoroughly evaluated. The measurements were conducted at 9 different elevations between 1710 and 2500 m from August 2014 to August 2015 with HOBO Data loggers. The results showed that the annual temperature lapse rates(TLRs) for Tmean,Tmin and Tmax were 0.45?C/100 m,0.44?C/100 m and 0.40?C/100 m,respectively,which are substantially smaller than the often used value of 0.60°C/100 m to 0.65°C/100 m. The TLRs showed no obvious seasonal variations,except for the maximum temperature lapse rate,which was steeper in winter and shallower in spring. Additionally,the TLRs showed significant diurnal variations,with the steepest TLR in forenoon and the shallowest in early morning or late-afternoon,and the TLRs changed more severely during the daytime than night time. The accumulated temperature above 0°C,5°C and 10°C(AT0,AT5 and AT10) decreased at a lapse rate of 112.8?C days/100 m,104.5?C days/100 m and 137.0?C days/100 m,respectively. The monthly and annual mean diurnal range of temperatures(MDRT and ADRT) demonstrated unimodal curves along the elevation gradients,while the annual range of temperature(ART) showed no significant elevation differences. Our results strongly suggest that the extrapolated regional TLR may not be a good representative for an individual mountainside,in particular,where there are only sparse meteorological stations at high elevations.     

Reproductive rate of a marine planktonic copepodLabidocera euchaet a Giesbrecht in Xiamen Harbor

Reproductive rate of a marine planktonic copepodLabidocera euchaeta Giesbrecht in Xiamen Harbor bor was studied during the period from April 1986 to August 1987. Results showed that rate of egg production (F, eggs/(♀, d)) was positively correlated with temperature (T,^oC) and body weight of spawning female (Wc, μgC) as:F=0.0637 (T−7)^0.7445 Wc. Each month a maximum rate of egg production occurred at one of the five experimental temperatures (10°, 15°, 25°, and 30°C) near the monthly mean field temperature. The available food density (P, μgC/ml) had significant effect on specific rate of egg production (Fc, %C ♀/d) as:Fc=0.1732 (P-0.0179)/P. The gross efficiency of egg production of ca 62% in carbon was independent of food density snd ingestion rate. Observations on in situ egg production and hatching rate demonstrated that this species reproduces continuously all the year round, with summer as its high reproductive season and winter a low one. Ten generations a year is also suggested based on the number of the peaks of average population egg production rate. This agreed well with the result obtained from the generation-time equations. Egg-laying pattern was also observed and discussed.