Ontogeny of digestive tract functionality in Japanese flounder, Paralichthys olivaceus studied by in vivo microinjection: pH and assimilation of free amino acids

To specifically study the functional ontogeny of the digestive tract, larvae of Japanese flounder at various developmental stages were injected with liquid solutions using tube feeding in vivo. The survival in the stages tested was on average 93%. The injected solutions were almost completely transferred from the presumptive stomach, or stomach to the midgut within 10 min of injection. The passage to the hindgut in some cases started 10 min after injection and over 90% of the solution had passed from the midgut to the hindgut after 1 h. In most cases the hindgut seemed to be completely empty after 3 h.Two different mixtures of pH indicators with sensitivities in the alkaline (7.5 to 9) and acid (4 to 6) ranges respectively were used for assessment of pH in the various gut segments. The pH of the stomach remained alkaline during the larval period, but had fallen close to 4 during late metamorphosis, an indication of active HCl secretion and progressive stomach differentiation. In mid and late metamorphosing fish a rapid colour change in the pH indicator was observed once it had passed the pyloric sphincter. This demonstrates that there was also active secretion of alkaline fluid, most likely HCO₃ ^–, from the pancreas into the pylorus lumen.A single injection of liquid solution of ¹⁴C-FAA showed that assimilation of FAA was high in all stages tested (79.5 ± 7.1%; SD; n=91). The presently reported data for Japanese flounder support earlier studies that FAA are absorbed with a high efficiency in the early stages of marine fish.     

Can fermented soybean meal and squid by‐product blend be used as fishmeal replacements for Japanese flounder (Paralichthys olivaceus)?

An experiment was carried out to evaluate fermented soybean meal and squid by‐product blend (1:1) (FP) as replacement of fishmeal (FM) for Japanese flounder, Paralichthys olivaceus. Five isocaloric (19 kJ g^?1) diets were prepared by replacing 0 (FP0), 12 (FP12), 24 (FP24), 36 (FP36) and 48% (FP48) FM protein with FP. Triplicate groups of juveniles (mean weight of 3.9 g) were delivered the test diets for 8 weeks in a flow‐through sea water system. The results showed that there were no significant differences (P > 0.05) among the growth rates of fish fed FP0, FP12, FP24 and FP36 diets. Growth and nutrient utilization parameters were significantly reduced in fish fed FP48 diet. Although, whole body proximate composition of fish were not significantly affected by the dietary treatments compared to the control; methionine and phenylalanine contents were significantly decreased in FP48 group. Protein retention was also significantly decreased in the similar group of fish. Dietary treatments did not alter most of the plasma metabolites, while some of the health parameters were improved in the replacement groups. Results suggested that FP is a potential candidate for alternative protein ingredient in aquafeed and can replace 36% FM protein in the diet of Japanese flounder.     

Characterization of the sex differentiation and gonadal development in small yellow croaker (Larimichthys polyactis) and its hybrid (L. polyactis ♀ × L. crocea ♂)

Fish Physiology and Biochemistry - Interspecific hybridization has been considered as a possible approach to improve biological traits and has been applied in aquaculture practices. In the present...     

Effects of dietary carnitine and lipid on growth and body composition of hybrid striped bass (Morone chrysops ♀× M. saxatilis ♂)

A study was undertaken to investigate the effects of graded dietary levels and different types of carnitine on hybrid striped bass (Morone chrysops × M. saxatilis %) fed different levels of lipid. An incomplete factorial design was utilized in which diets containing lipid at either 5 or 10% were supplemented with l-carnitine at 0, 500, or 1000 mg kg^–1 diet, dl-carnitine at 1000 mg kg^–1 diet, or carnitine chloride to provide 1000 mg carnitine kg^–1 diet. Juvenile hybrid striped bass (3.3 g fish^–1) were stocked into individual 38-l aquaria connected as a brackish water (6), recirculating system and fed each diet in triplicate for 9 weeks.Supplementation of the diet with 1000 mg carnitine kg^–1 increased muscle carnitine from 35.5 to 47.7 g g^–1 tissue. Carnitine supplementation did not result in increased weight gain regardless of carnitine level or type; however, weight gain showed a significant (p<0.05) response to dietary lipid with fish fed diets containing 10% lipid growing 34% more than fish fed diets with 5% lipid. The hepatosomatic index also was unaffected by diet, but the intraperitoneal fat (IPF) ratio was significantly elevated (5.1 vs 3.2%) in fish fed diets with 10% lipid compared to those fed diets with 5% lipid. Fish fed diets containing 1000 mg carnitine kg^–1 had increased IPF ratio values at 4.7% compared to 3.9% for fish fed the basal diet. Liver lipid also was responsive to dietary treatment, increasing from 6.7 to 8.8% of wet weight as dietary lipid increased from 5 to 10%. The relative quantities of triglycerides, free fatty acids and phospholipids in muscle and liver were not influenced by carnitine level, carnitine type or dietary lipid level. Supplementation of carnitine does not appear to be beneficial to hybrid striped bass based on either growth performance or body composition.     

Effects of total replacement of dietary fish oil on growth performance and fatty acid compositions of hybrid tilapia (Oreochromis niloticus × O. aureus)

The aim of this study is to evaluate the effects of total replacement of fish oil (FO) with pork lard (PL), soybean oil (SO), or tea seed oil (TO) on growth and fatty acid compositions of hybrid tilapia (Oreochromis niloticus × O. aureus). Four diets were formulated to contain 4.5 % of each of the different lipid sources. Each diet was assigned to triplicate groups of 15 hybrid tilapia with the same initial body weight (4.6 ± 0.5 g). After a 10-week feeding, the FO, SO, and TO groups had similar final body weight. The mean hepatosomatic index in the FO group was significantly lower than the other groups (2.5 vs 2.9–3.0; P < 0.05). Whole-body lipid levels were highest in the TO group (5.2 ± 0.4 %) and lowest in the FO group (4.3 ± 0.5 %). Total n-3 polyunsaturated fatty acids (PUFA) concentrations were greater in the SO and FO groups (5.5 ± 0.1 and 7.1 ± 0.2, respectively) than in the PL and TO groups (1.7 ± 0.1 and 1.4 ± 0.1, respectively). The FO group had a 2- to 4-fold reduction in the n-6/n-3 PUFA ratio relative to the other groups. The concentrations of both docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) were significantly (P < 0.05) higher in the FO group than in the other groups. Our data demonstrate that total replacement of dietary FO with SO or TO has little effects on growth of hybrid tilapia but reduces the contents of health-benefiting DHA and EPA in fish body.     

Interactions of Carbohydrate and Lipid in Diets for Hybrid Striped Bass (Morone chrysops × M. saxatilis)

The effects of changing the proportion of non-protein energy sources in diets for hybrid striped bass (Morone chrysops × M. saxatilis, Walbaum) were evaluated. Lipid (menhaden fish oil) and carbohydrate (wheat flour/dextrin 1:l) were varied at two digestible energy (DE) levels (2.9 kcal/g and 3.3 kcal/g) using a constant protein level of 35%. Fish fed at the lower energy level had significantly higher weight gain and protein efficiency ratios (PER) when lipid was 15% with no added carbohydrate. Fish fed at the higher energy level. however, exhibited signifcantly higher weight gain, apparent protein retention, and PER at 10% lipid and 30% added carbohydrates. Growth on this diet was also significantly better than other diets at both energy levels.     

Effects of chronic exposure to γ-HCH (Lindane) on brain serotonergic and gabaergic systems,and serum cortisol and thyroxine levels of rainbow trout,Oncorhynchus mykiss

Rainbow trout (Oncorhynchus mykiss) were implanted intraperitoneally with 0.5 ml.100 g^–1 body weight of coconut oil alone (controls) or coconut oil contaning 0.05 mg of -HCH (Lindane). After 18 days, changes in brain serotonin and GABA metabolism, as well as in serum cortisol and thyroxine levels, were measured. A lower final body weight was observed in -HCH treated fish when compared with control fish. No significant differences were found for serum thyroxine levels between control and treated fish, but a significantly higher cortisol level was found in the -HCH-implanted trout. Although GABA levels did not differ significantly in any brain region in the two treatment groups, the activity of the serotonergic system was significantly altered by the pesticide in both the hypothalamus and the telencephalon.     

Adapting bioenergetic factorial modelling to understand the implications of heat stress on barramundi (Lates calcarifer) growth,feed utilisation and optimal protein and energy requirements – potential strategies for dealing with climate change?

The implications of temperature on bioenergetics for barramundi (Lates calcarifer) were defined in an improved factorial model that encompassed revised parameters accounting for effects over the temperature range of 16–39 °C and size range of 10–3000 g. A revised growth function describing weight gain by barramundi as a function of fish weight and temperature was derived from farm and laboratory data and included a term for a shift in optimal temperature with fish size: Gain (g fish^?1 day^?1) = (K + xT + yT ² + zT³) * (weight)^ax+b. Maintenance energy and protein demand functions were also derived on a similar form, and all three functions combined to form the basis of a factorial model for energy and protein demand. Using this model, optimal iterative feed specifications were defined for a range of fish sizes at temperatures of 25, 30 and 35 °C. A feed demand model was also developed based on the demand for digestible energy (DE) at each of these temperatures. The model shows that at high temperatures (35 °C), there is an increase in digestible protein (DP) to DE demand, and that with increasing size, there is a decrease in the DP to DE demand.     

Distinct expression of three estrogen receptors in response to bisphenol A and nonylphenol in male Nile tilapias (Oreochromis niloticus)

Environmental estrogens, such as bisphenol A (BisA) and nonylphenol (NP), have been shown to affect the estrogen receptor (ER) expression and induce male reproductive abnormalities. To elucidate molecular mechanisms of action of xenoestrogenic chemicals on the expression of estrogen receptors in the testes of Nile tilapia (Oreochromis niloticus), three full-length cDNAs respectively encoding ntERα, ntERβ1 and ntERβ2 were cloned from testes. The amino acid sequences of ntERα, ntERβ1 and ntERβ2 showed a high degree of similarity to the relevant fish species. Tissue-specific expression study showed that three receptors were highly expressed in pituitary, liver, testis, kidney and intestine tissues. The ntERα, ntERβ1 and ntERβ2 mRNA expressions were significantly higher at the sexual early recrudescing stage than at other recrudesced stages. After being exposed to xenoestrogens from weeks 2 to 4, the ntERα mRNA levels were increased significantly in testes after NP treatment at all sampling times or after 4 weeks of exposure to BPA. The ntERβ1 mRNA levels remained unchanged, while a significant decrease of the ntERβ2 mRNA level was observed in testes after exposure to NP and BPA. The present study demonstrates that the regulation of all three ntER subtypes in testes may act via different molecular mechanisms of exposure to NP and BPA.     

Effects of Cottonseed and Alfalfa Meals as Fertilizers on Production of Palmetto Bass (Morone saxatilis × M. chrysops) Fingerlings in Plastic-Lined Ponds

Effects of cottonseed meal versus alfalfa meal fertilization on water quality, zooplankton density, and production of phase I palmetto bass, Morone saxatilis × M. chrysops, were compared in plastic-lined ponds. Cottonseed meal promoted better (P < 0.05) water quality, timing of high zooplankton density development, and growth and yield of palmetto bass. Cost of cottonseed meal toward fish production was $0.75/kg of fish, while that of alfalfa meal was $1.74/kg of fish.     

Oxygen consumption and ammonia excretion of black carp (Mylopharyngdon piceus Richardson) and allogynogenetic crucian carp (Carassius auratus gibelio ♀?×?Cyprinus carpio ♂) fed different carbohydrate diets

Oxygen consumption and ammonia excretion of black carp (Mylopharyngdon piceus Richardson) (4.6 ± 0.3 g) and allogynogenetic crucian carp (Carassius auratus gibelio ♀ × Cyprinus carpio ♂) (5.7 ± 0.5 g) were examined when fish fed two types of carbohydrate (dextrin and glucose) at two levels (20 and 40%) each. The diets were isonitrogenous (40% dry matter) and isocaloric at 18.5 kJ g⁻¹ (dry matter) by adjusting the oil content to 10.1 and 1.5%, respectively. In black carp, the interactions between the carbohydrate type and level were found in oxygen consumption at 3 and 6 h and in ammonia excretion at 6 h after feeding. At 20% carbohydrate, no significant difference was observed between dextrin and glucose in oxygen consumption. However, at 40% carbohydrate, oxygen consumption in fish fed glucose was significantly higher than that in fish fed dextrin at 3 and 6 h after feeding. Within the dextrin diets, no significant differences in both oxygen consumption and ammonia excretion were detected between the two carbohydrate levels. Within the glucose diets, however, fish fed 40% glucose showed significantly higher oxygen consumption than those fed 20% glucose at 3 and 6 h after feeding. Ammonia excretion in black carp fed 40% glucose was higher than that in black carp fed 40% dextrin at 6 h and also found higher than those in the other three treatments at 24 h after feeding. The postprandial oxygen consumption and the ammonia excretion in crucian carp fed 40% glucose were the highest, but no significant differences were observed. Our data indicate that the escalation of glucose to 40% in a fish diet results in high oxygen consumption and ammonia excretion in black carp, suggesting that the efficiency of glucose as an energy source for this fish is compromised by the high metabolic expenditure after feeding. Crucian carp, on the other hand, have a better ability to cope with dietary carbohydrates.     

Histological changes in insulin-immunoreactive pancreatic β-cells,and suppression of insulin secretion and somatotrope activity in brook trout (Salvelinus fontinalis) maintained on reduced food intake or exposed to acidic environment

Immature brook trout (Salvelinus fontinalis) were randomly divided into a pH control, a pH and food control and an acid-stressed group. Fish in the first two groups were held at neutral pH and those in the last group were maintained at pH 4.2 for up to two months. The food supply to the pH and food control group was restricted to simulate the reduction in food intake demonstrated for acid-stressed trout. Plasma insulin levels were significantly decreased from 5–20 ng/ml to 1–2 ng/ml and plasma cortisol levels were significantly increased from 5–10 ng/ml to as high as 70 ng/ml in the acid-stressed brook trout. Concomitantly, a significant decrease of 21–39% in the proportion (volume density) of insulin immunoreactive -cells was observed within the principal pancreatic islets. Somatic growth was stunted and ultrastructural morphometry revealed the suppression of somatotrope secretory activity in the acid-stressed fish. Restriction of food supply induced a smaller but still significant decrease in circulating levels of insulin which was however not accompanied by a reduction in insulin immunoreactive -cells. The rise in plasma cortisol levels was not significant, and the plasma levels of glucose and protein were unaffected. Nevertheless, somatotrope secretory activity was suppressed and somatic growth was stunted. This study demonstrates for the first time the complexity of the endocrine response to acid stress and that some of the response to acid stress can be attributed to the lowering of food intake.     

Growth performance of hybrid striped bass (Morone chrysops × M. saxatilis) fed with commercial pike perch and trout diets

Two commercial trout diets (Oncorhynchus mykiss) and one commercial pike perch diet (Sander lucioperca) were fed to hybrid striped bass (Morone chrysops × M. saxatilis) (mean initial weight ± SD of 60.7 g ± 12.1; mean initial length SD of 17.2 cm ± 1.1) for 69 days at rations of approximately 1% average body weight. While final body weight (FBW), final length (FBL) and condition factor (Cf) were not significantly influenced by diets, specific growth rate (SGR) in hybrid striped bass fed with the pike perch diet (1.15) was significantly higher than those fed with either of the two trout diets (1.04 and 1.07). The feed conversion ratio (FCR) in hybrid striped bass fed with the pike perch diet (1.0) was significantly lower than the FCR in hybrid striped bass fed with either of the two trout diets (1.1 and 1.2). When hybrid striped bass (mean initial body weight: 65.7 ± 4.5 and 127.7 ± 2.9 g) were fed with the pike perch diet twice per day until satiation for 52 days, the SGR was 1.7 and 1.15% d^?1 in fishes with an average body weight of 116 and 183 g, respectively. Present results demonstrate that growth performance in hybrid striped bass can be improved when fishes are fed with commercial pike perch diets rather than using commercial trout diets as is the current practice.

     

The effects of gonadal development and sex steroids on growth hormone secretion in the male tilapia hybrid (Oreochromis niloticus × O. aureus)

Profiles of plasma growth hormone (GH) in male tilapia hybrid (Oreochromis niloticus x O. aureus) were measured and compared at different times of the year. The profiles did not appear to be repetitive, however, differences in their nature were observed at the different seasons; the most erratic profiles were seen in the height of the reproductive season (July), while the peaks were more subdued in the spring and disappeared in the autumn. Peaks in male fish were more prominent than in the females when measured in July. Perifused pituitary fragments from fish with a high GSI responded to salmon gonadotropin-releasing hormone (sGnRH) analog (10 nM-1 M), while those from fish with a low GSI barely responded to even the highest dose. Exposure of perifused pituitary fragments from sexually-regressed fish to carp growth hormone-releasing hormone (cGHRH; 0.1 M) or sGnRH (I M) stimulated GH release only after injection of the fish with methyl testosterone (MT; 3 injections of 0.4 mg kg ¹). The same MT pretreatment did not alter the response to dopamine (DA; 1 or 10 M). GH pituitary content in MT-treated fish was lower than in control fish, which may be explained by the higher circulating GH levels in these fish, but does not account for the increased response to the releasing hormones. Castration abolished the response of cultured pituitary cells to sGnRH (I fM-100 nM) without altering either their basal rate of secretion or circulating GH levels. Addition of steroids to the culture medium (MT or estradiol at 10 nM for 2 days) enabled a GH response to sGnRH stimulation in cells from sexually regressed fish. Pituitary cells which had not been exposed to steroids failed to respond to sGnRH, although their response to forskolin or TPA was similar to that of steroid-exposed cells. It would appear, therefore, that at least one of the effects of the sex steroids on the response to GnRH is exerted proximally to the formation of cAMP, or PKC, presumably at the level of the receptor. An increase in the number of receptors to the GH-releasing hormones, following steroid exposure, would explain also the changing nature of the GH secretory profile in different stages of the reproductive season.     

Effect of starvation on activities and mRNA expression of lipoprotein lipase and hormone-sensitive lipase in tilapia (<Emphasis Type="Italic">Oreochromis niloticus</Emphasis> × <Emphasis Type="Italic">O. areus</Emphasis>)

A 4-week study was conducted to determine the effect of starvation on activities and mRNA expression of lipoprotein lipase (LPL) and hormone-sensitive lipase (HSL) in hybrid tilapia (Oreochromis niloticus × O. areus). The tissue samples were sampled once a week. Results showed that body weight (BW) and hepatosomatic index (HSI) were decreased significantly (P < 0.05) during starvation. The percentages of crude fat and crude protein in the whole body and the fat content in muscle decreased significantly (P < 0.05), while the rate of moisture and crude ash increased significantly (P < 0.05). The response of LPL, HSL activities and mRNA expression in tissues was tissue dependent. The activities of LPL and HSL in muscle at day 7 were elevated by 2.5 times (P < 0.05) and 11.8 times (P < 0.05) of the value at day 0, respectively, and both then decreased to pre-starvation levels at day 14 and finally stabilized at a certain level afterward. LPL and HSL mRNA abundance in muscle remained relatively stable between 0 and 14 day; then, a significant increase was seen after 14 days. In the liver, LPL activity maintained a significantly increasing trend during starvation, while HSL activity rose dramatically at day 7 of starvation by 2.35 times (P < 0.05) and finally stabilized at a certain level. The mRNA abundance of liver LPL increased significantly during the whole process of starvation (P < 0.05), whereas the mRNA abundance of liver HSL decreased significantly at day 7 of starvation, elevating significantly afterward (P < 0.05).