Use of neuroactive catecholamines to chemically induce metamorphosis of hatchery-reared flat oyster, Ostrea angasi, larvae

Low numbers and unreliable wild catch of the native flat oyster, Ostrea angasi , spat has resulted in the NSW flat oyster industry being reliant on hatchery-produced spat. The need to produce culchless spat in the hatchery stimulated investigation of several catecholamines to induce metamorphosis in O. angasi larvae. Larvae were treated with one of four neuroactive catecholamines (epinephrine, epinephrine bitartrate, l -Dopa and GABA) at one of four concentrations (10⁻³, 10⁻⁴, 10⁻⁵ or 10⁻⁶  m ) for one of three treatment durations (0.5, 1–2 h) to determine morphogenic action for culchless spat production. Epinephrine bitartrate at 10⁻³ and 10⁻⁴  m and epinephrine at 10⁻³, 10⁻⁴ and 10⁻⁵  m , for a treatment duration of 1–2 h, produced significantly greater numbers of spat and culchless spat, compared with any other treatment combination. The other catecholamines tested did not induce a significant increase in the total number of spat or culchless spat, over untreated controls. Separate trials found that long-term treatment (24 h) with epinephrine bitartrate and epinephrine at morphogenic concentrations inhibited metamorphosis. Consecutive daily use of epinephrine bitartrate increased the numbers of spat and culchless spat produced, but did not affect larval or short-term post-larval survival. Treatment with 10⁻³  m epinephrine bitartrate or 10⁻⁴  m epinephrine for 1 h is recommended for routine commercial production of culchless flat oyster spat.     

Gustatory receptor responses in marbled rockfish Sebastiscus marmoratus

Gustatory response of marbled rockfish Sebastiscus marmoratus was recorded from the facial nerve supplying the anterior palate. In amino acids, the lowest threshold was for L -proline at 10⁻⁶–10⁻⁵ M; in ATP-related substances, the value for inosine was approximately 10⁻⁵ M and in organic acids, the value for L -lactic acid was approximately 10⁻⁶ M. An almost complete self-adaptation was observed for stimulants after 5 s application. While a weak solution of L -lactic acid as adapting stimulus depressed the response to it completely, it did not appreciably affect the response to 10-times or more concentrated L -lactic acid solution. In contrast to a weak solution, a moderate solution caused a severe depression of the response to stronger solutions, resulting in a shift of the dose–response curve towards a high concentration range. Time for recovery from adaptation differed depending on chemicals and stimulus duration: for 10⁻⁴ M L -lactic acid, it took about 2.3 and 4.5 s for 1/2 recovery of response magnitude under unadapted condition from the adaptation after 1 s- and 5 s-stimulation, respectively. L -Lactic acid and L -alanine strongly depressed the response to each other, yielding a rightward shift of the dose–response curve. A similar profound cross-adaptation was also observed between inosine and 5'-inosinic acid. The possible role of gustation is discussed.     

Comparison of in vitro vitellogenin syntheses by three phenols in primary cultures of Chinese minnow, Rhynchocypris oxycephalus hepatocytes

The in vitro effects of bisphenol-A (BPA), nonylphenol (NP) and 4-tert-octylphenol (OP) on vitellogenin (VTG) synthesis were examined using primary cultures of Chinese minnow, Rhynchocypris oxycephalus hepatocytes. In vitro VTG syntheses were induced by exposure to 10^?5 M of BPA and 10^?4 M of both NP and OP, suggesting that BPA had the highest estrogenic potential among the three phenols.     

Perception of light intensity by Haliotis discus discus based on locomotor activity patterns

ABSTRACT:   An apparatus to measure the locomotor activity of aquatic benthic organisms at variable low light levels was developed and the diurnal behavioral pattern of the abalone Haliotis discus discus was measured at various low light intensities. During the experiment, abalone were exposed to 12 h light–dark cycles of complete darkness, 0 µmol/m²/s throughout the 12 h dark cycle and, during periods I (days 1–8) and III (days 19–26), the 12 h light cycles were set at 10 µmol/m²/s. During period II (days 10–17), abalone were exposed to a light level during the 12 h light cycles of 1 × 10⁻⁵, 1 × 10⁻⁶, 1 × 10⁻⁷ or 1 × 10⁻⁸ µmol/m²/s and the changes in locomotor activity assessed. At daytime levels of 1 × 10⁻⁵ µmol/m²/s, typical behavioral patterns were observed of high locomotory activity during the night-time cycle. However, at lower light intensities, the distinction between day and night activity patterns became less clear and, at intensities lower than 1 × 10⁻⁷ µmol/m²/s, the difference between activity during the light and dark cycles became negligible. Based on this, we conclude that the threshold of light level perception in relation to locomotor activity is approximately 1 × 10⁻⁷ µmol/m²/s. The significance of these results in relation to the entrainment of behavior in abalone is discussed.     

Underwater sound detection by cephalopod statocyst

Abstract:   The cephalopod receptor of particle motion was identified. In a previous study, it was suggested that statocysts served this function, but there was no direct supporting evidence, and epidermal hair cells had not been conclusively ruled out. Experiments on Octopus ocellatus were conducted using respiratory activity as an indicator of sound perception. Intact animals clearly responded to 141-Hz particle motion at particle accelerations below 1.3 × 10⁻³ m/s², and the mean perception threshold at this frequency was approximately 6.0 × 10⁻⁴ m/s². Specimens in which the statoliths had been surgically removed did not show any response for accelerations up to 3.9 × 10⁻³ m/s² at 141 Hz, which was approximately 16 dB greater than the mean perception threshold at this frequency. Specimens that had undergone a control operation in which the statoliths remained intact showed positive responses at 2.8 × 10⁻³ m/s² for the same frequency stimulus. This indicates that the statocyst, which is morphologically similar to the inner ear system in fish, is responsible for the observed responses to particle motion in O. ocellatus . This is the first direct evidence that cephalopods detect kinetic sound components using statocysts.     

Effect of Genistein on the Growth and Reproductive Function of Male and Female Yellow Perch Perca flavescens

We studied the effect of genistein, a weak estrogenic compound found in soybean products, on the growth and reproductive development of yellow perch Perca flavescens . Four diets were prepared: control, estradiol-17β (E₂) at 10 μg/g diet (E10), and genistein at 7.5 and 0.75 mg/g diet (G7.5 and G0.75). Fish (initial weight = 13–15.5 g and initial total length = 105–111 mm) were fed to excess for 63 d. The E10 diet promoted weight gain in yellow perch of both sexes, whereas the G7.5 diet decreased weight gain in females. The growth of the G0.75 group was not different from either the E10 group or the control group. Among females, fish fed the E10 diet had the lowest plasma concentration of E₂ 36 h after the last feeding of the study. The plasma concentration of E₂ did not differ among the males from the four treatments. The E10-treated fish of both sexes had increased plasma concentration of alkali-labile phosphoprotein phosphorous (ALPP) in both sexes, indicating that vitellogenesis had been induced. Neither genistein diet increased plasma (ALPP) concentration. Male but not female yellow perch fed the E10 diet had significantly lower gonadosomatic indices compared to control and genistein groups. These results suggest that genistein may have a positive effect on growth in yellow perch, but no apparent estrogenic effects on reproductive function.     

Larval settlement of the tropical abalone, Haliotis asinina Linnaeus, using natural and artificial chemical inducers

Many kinds of chemical and biological materials have been used as inducers of settlement of abalone larvae, as well as other species of marine gastropods, with responses being highly variable, even to the same chemical cue. The present study tested chemical inducers, γ-aminobutyric acid (GABA), δ-aminovaleric acid (5-AVA) and l -glutamic acid (GA) and the effects they have on larval settlement of Haliotis asinina . Additionally, a relatively inexpensive commercial substance, monosodium glutamate (MSG), was trialed. The datum provided shows all chemicals to be active inducers of settlement in this study, in order of effectiveness of 5-AVA, GABA, MSG to GA. Induction as adjudged from larval numbers settled was best at 6 h 62%, with 10⁻¹ mM 5-AVA. At 24 h, induction was the highest at 78% when exposed to 10⁻² mM 5-AVA. Larvae that were allowed to settle up to 72 h showed the highest numbers of settled larvae, and declined back to 60% when exposed to 10⁻² 5-AVA and 10⁻¹ mM GABA respectively. Monosodium glutamate, although third in settlement standings would bypass the other chemicals, with regard to cost versus yield. The assessment of settlement surface, rough or smooth proved to be irrelevant, which had no significant impact on larval settlement.     

The Annual Reproductive Cycle of the White Bass Morone chrysops

The gametogenic cycle of the white bass Moronc chrysops was characterized for one year at monthly intervals by measuring circulating levels of the sex steroid hormones testosterone (T) and 11-ketotestosterone (11-KT) in males, and 17β-estradiol (E₂) and T in females. Vitellogenin (VTG), the egg yolk precursor, was also measured in female blood plasma. Gonadal status of individual fish was assessed by histological evaluation, measurement of oocyte diameters, and spermintion response. At the onset of vitellogenesis (October), blood plasma levels of E₂ and T were low, but increased significantly as the spawning period (March-April) approached and oocytes attnined their maximum diameter. All stages of oocyte development were observed in mature females, indicating that the white bass is a multiple-clutch, groupsynchronous spawner. Androgen levels increased in males throughout the reproductive cycle and reached maximal levels during the spawning period. The increase in androgens coincided with sperm production and spermintion in males. The circannual endocrine and gonadal development patterns observed in white bass were similar to those of other members of genus Moronc .     

Correlates of growth in juvenile three-spined sticklebacks: potential predictors of growth rates in natural populations

Abstract - The possibility of using indices to quantitatively predict short-term growth rates of juvenile three-spined sticklebacks ( Gasterosteus aculeatus L.) was assessed. A range of weight-specific growth rates ( G _W) was obtained in experiments with individual sticklebacks fed enchtraeid worms at different daily rations over 21 days. There was a strong, positive correlation between G _W and the RNA : DNA ratio in white muscle ( r ²  = 0.90) and lipid concentration (as percentage dry weight) of the carcass ( r ²  = 0.92). There were smaller, but significant correlations between G _W and the percentage of dry matter in the carcass ( r ²  = 0.67), and the residuals from the weight–length relationship ( r ²  = 0.49). Regressions relating growth to RNA : DNA ratio and lipid concentration offer a means of estimating short-term growth rates in natural populations. A test of such predictions using the results from a 56-day experiment on stickleback growth suggested that predictions from percentage lipid were biased and inefficient, while predictions from the RNA : DNA ratio were unbiased but inefficient. Predictions from percentage lipid were higher than from the RNA : DNA ratio.     

Effects of dietary organic acids on growth, nutrient digestibility and gut microflora of red hybrid tilapia, Oreochromis sp., and subsequent survival during a challenge test with Streptococcus agalactiae

A 14-week feeding trial was conducted to determine the effects of dietary organic acids. The experimental diets were added with 0, 1, 2 or 3 g kg⁻¹ of a novel organic acid blend or with 2 g kg⁻¹ of potassium diformate and fed to triplicate groups of red hybrid tilapia ( Oreochromis sp.). Upon completion, tilapia were challenged by immersion with Streptococcus agalactiae . There was no significant difference ( P >0.05) in the growth, feed utilization and nutrient digestibility among treatment groups despite a trend towards improved results with fish fed organic acid-supplemented diets. Diet pH decreased, causing a reduction in the digesta pH of the stomach and gut. Total bacteria per gram of faeces were significantly ( P <0.05) reduced from 1.81 × 10⁸ colony-forming units (CFU) (control group) up to 0.67 × 10⁸ CFU in the fish fed organic acid diets. A similar trend was observed for adherent gut bacteria. Cumulative mortality of fish fed no organic acids was higher compared with fish fed organic acid-supplemented diets at 16 days post challenge. The data showed that dietary organic acids can exert strong anti-microbial effects and have the potential to exert beneficial effects on growth, nutrient utilization and disease resistance in tilapia.     

Detection of transgenic DNA in tilapias (Oreochromis niloticus, GIFT strain) fed genetically modified soybeans (Roundup Ready)

We used nested-polymerase chain reaction (PCR) to detect Roundup Ready soybean in aquatic feeds and feeding tilapias. A template concentration of 10⁻¹⁰ g μL⁻¹ DNA solution could be detected with a dilute degree of 0.01%. Most (90.6%) of the aquatic feeds containing soybean byproduct included exogenous DNA segments. We also compared genetically modified (GM) soybean with non-GM soybean diets in feeding tilapias ( Oreochromis niloticus , GIFT strain) and examined the residual fragments (254 bp) of GM soybeans. Tilapias receiving GM soybean diets had DNA fragments in different tissues and organs, indicating that exogenous GM genes were absorbed systemically and not completely degraded by the tilapia's alimentary canal.     

Effects of estradiol-17β and 17α-methyltestosterone on gonadal sex differentiation in the F2 hybrid sturgeon, the bester

The effects of giving oral estradiol-17β (E₂) and 17α-methyltestosterone (MT) on gonadal sex differentiation in the F₂ hybrid sturgeon, the bester ( Huso huso female ×  Acipenser ruthenus male), are investigated. Giving E₂ at 10 μg/g diet to fish from 14 months until 31 months of age induced incomplete feminization and resulted in approximately 40% abnormal ovary development in which oocytes were observed without ovarian lamellar structures and gonadal shape was similar to normal testis. Giving MT at 25 μg/g diet for the same duration failed to induce masculinization, and resulted in approximately 30% undeveloped gonads even at 30–37 months of age. In contrast, E₂ and MT at only 1 μg/g diet given from 3 to 18 months of age was sufficient to induce feminization and masculinization, respectively. In these fish, feminization and masculinization were observed at 9 months, when most putative ovaries and testes were histologically distinguishable by the shape of the gonadal surface. These results indicate that sex reversal can be induced in these fish by hormone treatment that is started at 3 months age, before morphological differentiation occurs on the stroma of the gonads.     

Comparison of horse mackerel and tilapia surimi gel based on rheological and <Superscript>1</Superscript>H NMR relaxation properties

ABSTRACT:   Horse mackerel and tilapia surimi were subjected to six different heat and pressure treatments in order to compare gelation characteristics of easy- and difficult-setting gels, that is, temperature dependence, by observing rheological properties and microscopic molecular mobility. The stress–relaxation and proton spin–spin relaxation time (¹H T ₂) of water were measured for all treated gels. Horse mackerel gel demonstrated higher elasticity, large distribution of the stress–relaxation process, and smaller water ¹H T ₂ than tilapia in both heat and pressure treatments. The water ¹H T ₂ was steeply increased in the pressure treatment at around 294 MPa for both fishes. In contrast, the ¹H T ₂ rarely changed in the heat treatment in spite of the considerable change in rheological properties. From the experimental results, it is considered that the gelation of horse mackerel (easy setting) surimi is induced by highly unfolding and re-aggregation of protein, which contributes to the formation of a strong network structure compared with tilapia in both heat and pressure treatments, and that pressure treatments hardly improve the gel strength of tilapia (difficult setting) surimi. The water ¹H T ₂ measurement was used effectively in order to study gelation characteristics of easy- and difficult-setting fish through observing its molecular dynamics.     

Seasonality of serum levels of vitellogenin in male Japanese whiting, Sillago japonica, reared under natural temperature and photoperiod

ABSTRACT:   Because blood vitellogenin (Vg) has been considered a biomarker for environmental estrogens, the basal levels of Vg and 17β-estradiol (E₂) were determined in male Japanese whiting reared under natural conditions. Serum levels of Vg and E₂ were measured and gonadal development was assessed by gonadosomatic index (GSI) and histological observation in 8–10 male fish at monthly intervals throughout the annual reproductive cycle. Serum E₂ was <60 pg/mL throughout the study period. In contrast, serum Vg exhibited seasonal changes: serum levels of Vg gradually increased from April to May (mean 63 ± 13 ng/mL and 124 ± 48 ng/mL in April and May, respectively), and then reached a peak value (mean 352 ± 68 ng/mL) in June. Thereafter, serum Vg gradually decreased, reaching undetectable levels (<50 ng/mL) in October. Serum levels of Vg tended to increase in the male fish in which the GSI was >1%. Histological observation revealed that testes in such male fish were in active spermatogenesis and then all of the testes of male fish in which serum Vg decreased to ND levels were regressed. These results suggest that Vg productive potency (sensitivity to estrogens) may increase in the spermatogenic stage, resulting in production of Vg in response to very low levels of natural or xenobiotic estrogens.     

Continuous exposure to infectious pancreatic necrosis virus during early life stages of rainbow trout, Oncorhynchus mykiss (Walbaum)

Rainbow trout, Oncorhynchus mykiss (Walbaum), were exposed continuously to infectious pancreatic necrosis virus (IPNV) at 0, 10¹, 10³ or 10⁵ plaque forming units (pfu) L⁻¹ of water to estimate the effects of chronic IPNV exposure on early life stages. Fish density averaged 35 fish L⁻¹ (low density) or 140 fish L⁻¹ (high density), and the tank flow rate was 250 mL⁻¹ min. Virus exposure began at 6 days before hatch and continued until fish were 44 days old. Cumulative per cent mortality, analysis of survival and hazard functions, and discrete-time event analysis were used to explore the patterns of survival and mortality. In eggs and fish exposed to IPNV, mortality significantly greater than in the 0 pfu L⁻¹ exposure did not occur until IPNV concentration was 10⁵ pfu L⁻¹ at low fish density and 10³ pfu IPNV L⁻¹ at high fish density. These results suggest that in the natural aquatic environment, where rainbow trout densities are likely to be considerably lower than in this study, mortality resulting from infection with IPNV will very likely not occur when ambient concentrations of virus are ≤10³ pfu IPNV L⁻¹. In aquaculture rearing units, trout density is likely to be as high or higher than the densities used in this study. Therefore, continuous inputs of virus at concentrations greater than 10¹ pfu L⁻¹ may result in IPN epidemics in aquaculture facilities.     

A pharmacokinetic study of flumequine in sea bass, Dicentrarchus labrax (L.), after a single intravascular injection

The pharmacokinetic properties of flumequine following a single intravascular injection (10 mg kg^–1 fish) were studied in sea bass, Dicentrarchus labrax (L.), 120 g held at 18 °C. The absorption half life ( t _1/2α) and the elimination half life ( t _1/2β) of the drug were calculated to be 1.05 and 10.71 h, respectively. Tissue penetration of flumequine seemed to be moderate because both the apparent volume of distribution of the drug at steady-state ( V _d(ss)) and the extensive apparent volume of the central compartment ( V _c) were found to be small (1.51 and 0.626 L kg^–1). The mean residence time ( MRT ) was short (09.73 h) and the total clearance (CL_T) of the drug was rapid (0.156 L kg^–1 h^–1).     

Nutritional evaluation of waste date fruit as partial substitute for soybean meal in practical diets of juvenile Nile tilapia, Oreochromis niloticus L.

The potential of waste date meal (WDM; low-quality date palm, Phoenix dactylifera L.) as a carbohydrate source in formulated diets for Nile tilapia was evaluated. Four isocaloric-practical diets (15.7 kJ g⁻¹) were formulated incorporating WDM at 0, 100, 200 and 300 g kg⁻¹ levels as partial substitutes for soybean meal (SBM). These were designated D₀ [284 g crude protein (CP) and 383 g carbohydrate (CHO) kg⁻¹ diet], D₁ (279 g CP and 446 g CHO kg⁻¹ diet), D₂ (207 g CP and 495 g CHO kg⁻¹ diet) and D₃ (175 g CP and 578 g CHO kg⁻¹ diet). Each diet was fed to three replicate groups of 30 fish [20.20 ± 0.09 g (±SE)] for 75 days. No feed-related mortality was observed during the entire experimental period. Final body weight (FBW) and specific growth rate (SGR) in the different treatments were statistically not significantly different ( P  > 0.05). Protein efficiency rate (PER) was lowest in diet D₀ and increased with decrease of SBM content (D₁–D₃). A significant increase in whole body lipid content was recorded in fish fed diets D₂ and D₃. Results showed that WDM could be a substitute for SBM up to 300 g kg⁻¹ in practical Nile tilapia diets without compromising growth.     

Migration of anadromous white-spotted charr <Emphasis Type="Italic">Salvelinus leucomaenis</Emphasis>, as determined by otolith strontium: calcium ratios

ABSTRACT:   The migratory history of anadromous white-spotted charr Salvelinus leucomaenis collected from Japanese coastal waters, was examined in terms of strontium (Sr) and calcium (Ca) uptake in the otolith, by means of wavelength dispersive X-ray spectrometry using an electron microprobe. Otolith Sr concentration or Sr : Ca ratios of anadromous S. leucomaenis , fluctuated strongly along the life history transect in accordance with the migration (habitat) pattern from sea to fresh water. The anadrmous S. leucomaenis showed phase L (low Sr : Ca ratio) from the core to the point 1000–2500 µm distant, averaging 1.3 × 10⁻³ to 2.7 × 10⁻³ and thereafter, the ratios increased sharply, being higher than 5.0 × 10⁻³ to 10.0 × 10⁻³. These findings indicated that otolith Sr : Ca ratios reflected individual life histories, enabling a sea habitat to be identified from a freshwater habitat in this species.     

The assessment of the chemical composition of fishmeal by near infrared reflectance spectroscopy

The use of near infrared reflectance spectroscopy (NIRS) was investigated as an alternative method for predicting moisture (M), oil, crude protein (CP), ash, salt as NaCl, total volatile nitrogen (TVN) and buffer capacity in fishmeal. The NIRS calibration models were developed using the modified partial least squares (MPLS) regression technique. One thousand and ten ( n =1010) fishmeal samples were used to predict chemical composition for quality control in the fishmeal industry. Equations were selected based on the lowest cross validation errors (SECV). The coefficient of determination in calibration ( R ²) and SECV were 0.93 and 3.9 g kg^–1 dry matter (DM); 0.85 and 5.7 g kg^–1 DM; 0.92 and 3.7 g kg^–1 DM; 0.91 and 4.7 g kg^–1 DM; 0.88 and 6.7 g kg^–1 DM; 0.94 and 1.8 g kg^–1 DM; for M, CP, oil, ash, TVN and NaCl, respectively. It was concluded that NIRS can be used as a method to monitor the quality of fishmeal under industrial conditions.