Biological effects of insulin on murine melanoma cells and fish erythrophoroma cells: A comparative study

Insulin is the hormone that plays an essential role in metabolism and mitosis of normal and tumor cells, exerting its pleiotropic effects through binding to specific membrane receptors and promoting the phosphorylation of tyrosine residues of the receptor itself and of other components of the signaling pathway. The aim of this study was to investigate the effects of insulin on melanogenesis and cell growth in three different cell lines: the goldfish GEM-81 erythrophoroma cells (undifferentiated and differentiated with 1.5% dimethylsulfoxide-DMSO), and the murine B16F10 and Cloudman S91 melanoma cells. Undifferentiated GEM-81 and B16F10 cells responded to insulin with a small increase of cell proliferation, whereas S91 cells responded with a decrease of growth. In the two mammalian cell lines, and in DMSO-differentiated GEM-81 cells, the hormone strongly inhibited melanogenesis, by decreasing tyrosinase activity. In undifferentiated GEM-81 cells, insulin had no effect on tyrosinase activity. An increase in the tyrosine phosphorylation status of pp 185 (insulin receptor substrate 1 and 2-IRS-1/2) phosphorylation degree was observed in S91 mouse melanoma and in differentiated GEM-81 erythrophoroma cells, suggesting that this specific protein was maintained during transformation process and participates in insulin signaling. Our results imply an ancient and diverse history of the insulin signaling system in vertebrate pigment cells. (C) 2008 Elsevier Inc. All rights reserved.

Effect of Fish Oil Supplementation for Two Generations on Changes of Lymphocyte Function Induced by Walker 256 Cancer Cachexia in Rats

Fish oil supplementation has been shown to improve the cachectic state of tumor-bearing animals and humans. Our previous study showed that fish oil supplementation (1 g per kg body weight per day) for 2 generations had anticancer and anticachetic effects in Walker 256 tumor-bearing rats as demonstrated by reduced tumor growth and body weight loss and increased food intake and survival. In this study, the effect of fish oil supplementation for 2 generations on membrane integrity, proliferation capacity, and CD4/CD8 ratio of lymphocytes isolated from mesenteric lymph nodes, spleen, and thymus of Walker 256 tumor-bearing animals was investigated. We also determined fish oil effect on plasma concentration and ex vivo production of cytokines [tumor necrosis factor-alpha (TNF-alpha), interferon-gamma (IFN-gamma), interleukin-4 (IL-4), IL-6, and IL-10]. Lymphocytes from thymus of tumor-bearing rats presented lower viability, but this change was abolished by fish oil supplementation. Tumor growth increased proliferation of lymphocytes from all lymphoid organs, and fish oil supplementation abolished this effect. Ex vivo production of TNF-alpha and IL-6 was reduced in supplemented animals, but IL-4 and IL-10 secretion was stimulated in both nontumor and tumor-bearing rats. IL-10 and IFN-gamma plasma levels was also decreased in supplemented animals. These results suggest that the anticachetic effects of fish oil supplementation for a long period of time (2 generations) in Walker 256 tumor-bearing rats may be associated to a decrease in lymphocyte function as demonstrated by reduced viability, proliferation capacity, and cytokine production.

Glucose metabolism by lymphocytes, macrophages, and tumor cells from Walker 256 tumor-bearing rats supplemented with fish oil for one generation

Here we investigated the effect of lifelong supplementation of the diet with coconut fat (CO, rich in saturated fatty acids) or fish oil (170, rich in n-3 polyunsaturated fatty acids) on tumor growth and lactate production from glucose in Walker 256 tumor cells, peritoneal macrophages, spleen, and gut-associated lymphocytes. Female Wistar rats were supplemented with CO or FO prior to mating and then throughout pregnancy and gestation and then the male offspring were supplemented from weaning until 90 days of age. Then they were inoculated subcutaneously with Walker 256 tumor cells. Tumor weight at 14 days in control rats (those fed standard chow) and CO supplemented was approximately 30 g. Supplementation of the diet with FO significantly reduced tumor growth by 76%. Lactate production (nmol h(-1) mg(-1) protein) from glucose by Walker 256 cells in the group fed regular chow (W) was 381.8 +/- 14.9. Supplementation with coconut fat (WCO) caused a significant reduction in lactate production by 1.6-fold and with fish oil (WFO) by 3.8-fold. Spleen lymphocytes obtained from W and WCO groups had markedly increased lactate production (553 +/- 70 and 635 +/- 150) when compared to non-tumor-bearing rats (similar to 260 +/- 30). FO supplementation reduced significantly the lactate production (297 +/- 50). Gut-associated lymphocytes obtained from W and WCO groups increased lactate production markedly (280 +/- 31 and 276 +/- 25) when compared to non-tumor-bearing rats (similar to 90 +/- 18). FO supplementation reduced significantly the lactate production (168 +/- 14). Lactate production by peritoneal macrophages was increased by tumor burden but there was no difference between the groups fed the various diets. Lifelong consumption of FO protects against tumor growth and modifies glucose metabolism in Walker tumor cells and lymphocytes but not in macrophages. Copyright (C) 2008 John Wiley & Sons, Ltd.

Growth, lipogenesis and body composition of piracanjuba (Brycon orbignyanus) fingerlings fed different dietary protein and lipid concentrations

Piracanjuba (Brycon orbignyanus) is a Brazilian migratory fast-growing omnivore, very appreciated as a sport fish, which is threatened to extinction in Southern Brazil due to stock over exploitation and dam building. Therefore, efforts have been made to raise this fish in captivity for reintroduction and aquaculture purposes. In the present study, the effects of different dietary protein and lipid concentrations on piracanjuba fingerlings growth performance, feed utilization, body composition, hepatosomatic index (HSI) and activity of the lipogenic enzymes fatty acid synthetase (FAS), glucose-6-phosphate dehydrogenase (G6PD) and malic enzyme (ME) were investigated using a 2 x 3 factorial experiment. Six casein-gelatin based diets were prepared combining two protein (30% and 32%) and three lipid concentrations (5.5%, 8.8% and 12.1%). Eleven fish, average weight 11.30 +/- 0.1 g, were held in each of 18 100-1 aquaria, supplied with recirculating freshwater. Each diet was randomly assigned to triplicate groups of fish and fed to apparent satiation, twice a day for 100 d. Piracanjuba fingerlings' daily weight gain (0.36-0.40 g), specific growth rate (1.43-1.51%), feed utilization and HSI were not influenced by dietary protein or lipid concentration. However, body composition was directly affected by dietary treatment. An increase in body fat and dry matter was observed as dietary lipid increased, for both dietary protein concentrations tested. The activity of FAS was depressed by increasing dietary fat levels but the G6PD activity did not differ among dietary treatments, although ME activity showed some regulation by dietary protein. These results indicate that an increase from 5.5% to 12.1% in the dietary lipid, at a dietary protein concentration of 30% or 32%, promotes body fat accumulation in piracanjuba fingerlings with no improvement in growth, suggesting that the lipid requirement for this species should be 5% or less, when raised for commercial purposes. However, the additional energy reserve from body fat accumulation could be desirable for piracanjuba fingerlings produced for stock enhancement. (C) 2003 Editions scientifiques et medicales Elsevier SAS and Ifremer/IRD/Inra/Cemagref. All rights reserved.

Free- and peptide-based dietary arginine supplementation for the South American fish pacu (Piaractus mesopotamicus)

Arginine was hypothesized to be a model compound in the present study on molecular forms of indispensable amino acid (IAA) dietary supplementation. Juvenile South American pacu (Piaractus mesopotamicus) were fed diets containing arginine in a protein base (casein-wheat gluten or casein-gelatin), or the casein-wheat gluten base supplemented with dipeptide or free arginine at two levels (5 and 10 g kg(-1)). Growth and protein efficiency ratios were significantly affected by diets, but not by arginine molecular form. Three free dispensable amino acids (DAA) and four IAA in plasma were affected by diet, but plasma arginine concentrations did not differ. Plasma urea concentrations, being very low in the pacu, and hepatic arginase activities, were not affected by diet (P = 0.10-0.11), but together with plasma ornithine, mirrored the growth data. Molecular form of arginine supplementation, free or dipeptide, significantly changed several free IAA (Phe, Leu, Ile, His) and urea, with a higher mean plasma concentration in dipeptide fed fish. The dietary treatments, or molecular form of the arginine supplementation, did not change proximate composition, except that calcium levels decreased with higher dietary arginine supplementation level. The present study indicates that dipeptides can provide IAA to pacu, and that arginine supplemented in this form is utilized as efficiently as in free form.

Growth performance and energetic metabolism of pacu, Piaractus mesopotamicus (Holmberg, 1887), fed diets supplemented with ammonium metavanadate

Vanadium compounds mimic most of the metabolic effects of insulin, suggesting that it might be useful to improve utilization of dietary carbohydrate. This work evaluated the effect of dietary ammonium metavanadate (H(4)NO(3)V) on the growth performance and energy metabolism of pacu, an omnivorous South America characin. Two hundred and eighty-eight fish were distributed into four blocks according to the body weight (21.8 +/- 1.7, 28.5 +/- 2.0, 28.4 +/- 1.9, 35.7 +/- 1.9 g), stocked in 24 plastic tanks and fed twice daily with isonitrogenous and isoenergetic diets containing six levels of H(4)NO(3)V (0, 10, 50, 100, 300 and 1000 mg kg(-1)) for 60 days. Increasing levels of dietary ammonium metavanadate did not improve growth (P > 0.05), and the highest level of inclusion (1000 mg kg(-1)) reduced performance (P < 0.05). Blood glucose levels decreased (P < 0.05) in fish fed 300 and 1000 mg kg(-1) H(4)NO(3)V, but no differences were observed in other blood metabolites. A slight increase in muscle lipid content was observed in fish fed a diet containing 300 mg kg(-1) H(4)NO(3)V. Based on the results of this study, there is no benefit in supplementing pacu diets with metavanadate.

Effects of photoperiod on reproduction of Siamese fighting fish Betta splendens

This study investigated the effect of different photoperiods (24 h of light (L):0 h of darkness (D); 20L:4D; 16L:8D; 12L:12D; and 8L:16D) on the reproduction and growth of Betta splendens. The results showed that spawning frequency was significantly higher in couples reared under 16L:8D and 12L:12D, in comparison with other treatments. The highest number of eggs per spawn was obtained under 16L:8D (544.76±375.23) and 12L:12D (471.13±261.52), and the lowest values were detected for 24L:0D (128.55±58.14) and 20L:4D (187.87±103.84). Fertility and fecundity also showed significantly higher average values in 16L:8D and 12L:12D when compared with 24L:0D and 20L:4D treatments. Egg volume and perivitelline space were significantly higher in 24L:0D treatments that showed the lowest numbers of eggs per spawn, while the vitelline volume did not show significant differences. Other variables such as breeders weight gain and condition factor (K) were not statistically different. Moreover, the final length varies according to photoperiod and gender. These results demonstrated a key role for the photoperiod upon B. splendens reproduction. The best reproductive performance is achieved under the photoperiods that best approached those that occur in spring and summer (16L:8D and 12L:12D), coinciding with their best seasons for reproduction.

Growth, fat content and fatty acid profile of South American catfish, surubim (Pseudoplatystoma fasciatum) juveniles fed live, commercial and formulated diets

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Effects of different dietary lipid sources on the survival, growth, and fatty acid composition of South American catfish, Pseudoplatystoma fasciatum, Surubim, juveniles

The present study examines the effect of four semi-purified diets (casein-gelatin based) where the source of fatty acids was free (esterified) oleic acid and linoleic acid (LA) (LOA diet), linseed and olive oil (predominantly LA and linolenic acid) (LO diet), cod liver oil (rich in highly unsaturated fatty acids) (CLO diet), and soybean lecithin (phospholipids; mostly LA) (LE diet) on the growth of juvenile South American catfish (surubim, Pseudoplatystoma fasciatum, Pimelodidae) (0.98 +/- 0.04 g individual weight). Fish were fed at a restricted-readjusted feeding rate for 8 wk. At the end of the experiment, LE-diet-fed fish grew significantly larger than those of the other three groups (P < 0.05). Considerable cannibalism was observed in all the treatments. It is suggested that the quantitative growth performance may possibly change under other conditions, with less or no cannibalism. Survival did not differ significantly among the fish fed four different diets. Muscle and liver lipid contents did not vary among dietary treatments (P > 0.05), but whole-body lipid concentrations were affected by dietary treatments. Fish fed LE diet contained significantly lower lipid level than those fed three other diets (P < 0.05). Muscle and liver fatty acid profiles reflected dietary fatty acid composition. Arachidonic acid level was significantly higher in muscle and liver of fish fed LOA and LE diets than in those fed LO and CLO diets. The results suggest that the efficiency of elongation and desaturation of 18C fatty acids depends on the dietary lipid source, and South American catfish has considerable capacity to transform linoleate to arachidonate.

Live and formulated diet evaluation through initial growth and survival of jundiá larvae, Rhamdia quelen

A larvicultura da maioria das espécies de peixes enfrenta o desafio da dependência do alimento vivo (AL) e da falta de dietas formuladas (DF) que atendam plenamente às necessidades das larvas. A baixa digestibilidade e a qualidade nutricional das DFs são alguns dos fatores que explicam o insucesso quando as larvas recebem apenas FD. Para avaliar o efeito da combinação da DF com o AL no crescimento e na sobrevivência de larvas de jundiá (Rhamdia quelen), comparando com o uso separado da DF ou do AL, larvas recém eclodidas (5,57 mm; 1,41 mg) foram estocadas inicialmente em 12 aquários de 10 L (100 larvas por aquário). Quatro réplicas foram alimentadas ad libitum com uma das três dietas por 20 (para DF) ou 48 dias (para AL ou a combinação DF + AL). As larvas alimentadas com apenas DF apresentaram crescimento e sobrevivência reduzidos quando comparadas àquelas alimentadas com AL ou a combinação DF + AL. Adicionalmente, as larvas do tratamento DF + AL apresentaram maior crescimento em peso (170 mg) que aquelas alimentadas apenas com AL (110 mg). O melhor desempenho das larvas alimentadas com DF + AL mostra que a maioria dos nutrientes exigidos pelas larvas é fornecida mais adequadamente quando ambas as dietas são fornecidas juntamente. Contudo, trabalhos sobre nutrição larval poderão contribuir ainda mais sobre a elucidação deste tema quando feitas comparações com o uso combinado de DF + AL, do que apenas testando isoladamente novos ingredientes e fontes protéicas normalmente utilizadas na elaboração de dietas para juvenis e adultos.

Haematological response and growth performance of Nile tilapia (Oreochromis niloticus L.) fed diets containing folic acid

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Growth performance and metabolic response of Nile tilapia fed rations supplemented with autolized yeast and zinc

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effects of incubation temperature on muscle morphology and growth in the pacu (Piaractus mesopotamicus)

This study describes the influence of incubation temperature during initial development phase on the morphology and muscle growth characteristics in the pacu (Piaractus mesopotamicus). Pacu eggs were incubated at 25, 27, and 29 degreesC until hatching. After day 5, fish from each temperature were transferred to 5001 tanks. At hatching and after 5, 25, and 60 days, muscle samples were collected, some were frozen in liquid nitrogen and others fixed in 4% paraformaldehyde or 2.5% glutaraldehyde. These samples were used for morphological, histochemical, immunohistochemical, and morphometric analysis. At hatching, we observed a superficial monolayer of small diameter fibers, lying just beneath the skin surrounding several round cells. From day 5, we observed two distinct populations of muscle fibers distributed in two layers: (1) red-in a superficial region with aerobic activity, and following acid preincubation, high mATPase activity, and 2) white-with anaerobic activity, and following alkaline preincubation, high mATPase activity. Twenty-five days after hatching, an intermediate layer and cell proliferating zones could be seen in the dorsal fin muscle region, with intermediate characteristics. Throughout the experimental period, there was an increase in muscle mass due to new fiber recruitment in the cell proliferating zones and between the more differentiated fibers in red, intermediate, and white muscles. This was more obvious from day 25, and at 29 degreesC than at 25 and 27 degreesC. Fiber hypertrophy occurred from hatching to 60 days and was more evident from 5 to 25 days. The number of proliferating nuclei (PCNA-labelling) increased from hatching to 60 days, and was more obvious in the 29 degreesC group at 60 days. Our results show that at incubation temperatures of 25, 27 and 29 degreesC, hypertrophy was predominantly from hatching to 25 days, after that muscle growth by hyperplastic mechanism increased. The interaction of muscle hypertrophic and hyperplastic growth processes in the 29 degreesC group produced the largest fish at the end of the experiment. (C) 2004 Elsevier B.V. All rights reserved.

Morphological and histochemical study of the myotomal muscle in pacu (Piaractus mesopotamicus Holmberg, 1887) during the initial growth phases

Morphological and histochemical methods were used to evaluate the myotomal muscle characteristics of pacu (Piaractus mesopotamicus) from hatching to 40 days old. During the larval period, the musculature consisted predominantly of white muscle. White and red muscle mass increased at 10, 20, 30 and 40 days after hatching. The larvae had round muscle fibers with a moderate degree of maturation and central nuclei. In subsequent phases, small and immature fibers were visible near larger and more differentiated fibers. Undifferentiated cells or presumptive myoblasts located in embryogenic zones were visible in the dorsal and ventral regions, and were more evident at 30 and 40 days. The red muscle fibers located in the subdermal region, had oxidative metabolism and slow contraction, whereas the more predominant white muscle fibers had glycolytic metabolism and fast contraction. Our findings indicate that during the initial phases, myotomal muscle growth in pacu occurs by both, muscle fiber hypertrophy and hyperplasia. The analysis of frequency of red and white muscle fibers shows that hyperplastic growth is intense in this period. As the growth rate in adult fish is related to the number of muscle fibers in young fish, extrinsic factors could change the muscle fiber phenotype and influence their ultimate size.

Growth characteristics of skeletal muscle tissue in Oreochromis niloticus larvae fed on a lysine supplemented diet

The effect of lysine amino acid supplementation on the growth characteristics and morphological pattern of skeletal muscle tissue in Nile tilapia Oreochromis niloticus larvae was evaluated. There were four treatments (T) with increasing levels of lysine supplement (T1 = 0.0%: T2 =1.1%; T3 = 1.7%; T4 = 4.0%) and one treatment with a commercial diet (T5). In all treatments, morphological and histochemical muscle tissue analyses were similar. Two distinct layers were identified: a superficial red layer, more developed in the lateral line region, formed by fibres with intense to moderate NADH-TR reaction and strong acid-stable mATPase activity, and a deep white one, most of the Muscle mass, formed by fibres with weak NADH-TR reaction and strong alkali-stable mATPase activity. There was an intermediate layer between these two layers with fibres exhibiting either weak acid-stable or acid-labile mATPase activity. Body mass increase was significantly higher in T5 than in the lysine treatments (T1-T4). There was no difference in number and diameters of muscle fibres between lysine treatments. In T5, muscle fibre diameter and number were higher. The frequency of red fibres with diameters <= 8 mu m was higher in the lysine treatments, and with diameters between 16 and 24 mu m, was higher in T5. Most white fibre diameters in T5 were significantly larger than 24 mu m and in T1-T4 were between 8 and 16 mu m. Cell proliferation was higher in the lysine treatments and muscle growth in T5 was mainly by fibre hypertrophy. (c) 2005 the Fisheries Society of the British