Effect of hypo- and hypersaline conditions on osmolality and Na+/K+-ATPase activity in juvenile shrimp (Litopenaeus vannamei) fed low- and high-HUFA diets.

Fatty acid composition of cellular membranes can modify permeability and can modulate the activity of Na(+)/K(+)-ATPase. Although highly unsaturated fatty acids (HUFA) improve survival and osmoregulatory capacity to low salinities in penaeid shrimp, the possible mechanisms have not been established. For this purpose the influence of HUFA supplementation in diet (2.9 vs. 34% HUFA proportion to total fatty acids) on osmoregulatory responses of juvenile Litopenaeus vannamei submitted to an acute (15 h) or chronic exposure (21 days), to low (5 g L(-1)) and high salinities (50 g L(-1)) was analyzed. Shrimp fed the high-HUFA diet, had higher concentration of main HUFA (20:5n-3 and 22:6n-3) in polar lipids of gills. Osmotic pressure in hemolymph was significantly affected by salinity in acute (640, 751, 847 mOsm/kg for 5, 30 and 50 g L(-1), respectively), and chronic exposure (645, 713, 814 mOsm/kg), but variations between them were small compared to environmental salinity (206, 832, 1547 mOsm/kg), indicating that osmoregulation was achieved in a matter of hours. An increase in Na(+)/K(+)-ATPase activity was observed only after a chronic exposure to low salinity. Free amino acids (FAA), mainly alanine and arginine, were higher at 30 (control) and 50 g L(-1) in accordance to their role as organic osmolites. Neither osmotic pressure, Na(+)/K(+)-ATPase activity, nor FAA was affected by HUFA supplementation. However, higher water content in gills of shrimp exposed to low salinities was counteracted by increased HUFA content, which could be a result of changes in water permeability of gills. The osmoregulatory capacity of penaeid shrimp to low and high salinities was achieved within 15 h of acclimation and did not depend on HUFA supplementation in the diet.

Comparative biochemical composition in gonad and adductor muscle of triploid and diploid catarina scallop (Argopecten ventricosus Sowerby II, 1842).

Biochemical components of gonad and adductor muscle for diploid and triploid catarina scallop, Argopecten ventricosus, were evaluated and compared at four periods in 1 year (January, April, June, and October). Two comparisons were done. The first one compared an untreated control (diploid) vs. a triploidy-treated group for which the percentage of triploids was 57%. The second comparison was done on a group derived from within the triploidy-treated group, separating diploids (internal control) from triploids ('putative triploids'). Regardless of which comparison, in the gonad diploid scallops had larger concentrations of proteins, carbohydrates, lipids, and acylglycerols than triploid scallops. This reflects the maturation processes in diploid scallops vs. the sterility seen in most triploid scallops, and it is particularly supported by the consistently larger concentration of acylglycerols in gonads of diploids than in triploids. The gonad index of the internal control (diploid) group was significantly larger than that seen in the putative triploid group at all sampling periods but October, when none of the gonad biochemical components were different between ploidy groups.Triploid scallops had a significantly larger muscle index than diploids from April to October. This can be caused by a larger gain in muscle tissue in triploids than diploids from January to June. However, there were no consistent differences in any of the biochemical components evaluated in adductor muscle of diploids and triploids. The use of freshly ingested food rather than reserve mobilization from muscle in diploids is suggested by these results. Nutrients derived from ingested food are apparently used for muscle growth in triploids, whereas in diploids those nutrients serve primarily for gonad development. The importance of freshly ingested food for maintenance and growth is suggested because the decline in biochemical components seen in October in both muscle and gonad was paired with a decline in weight of those two organs, especially when the control groups are considered, but a decrease was also evident for the triploid groups. This may have been caused by the presence of El Niño, with its characteristic high water temperatures and low productivity.

Metabolic responses of the white shrimp, Penaeus vannamei, to ambient ammonia.

Juvenile shrimp were individually exposed during 24 h to 0.007 (control), 0.36, 1.07, and 2.14 mmol/l total ammonia-N at 28 degrees C and 39 ppt salinity. After 22 h of ammonia-N exposure, oxygen consumption was measured for 2 h, and then hemolymph, hepatopancreas, and muscle tissues were sampled. Oxygen consumption, and levels of lactate and glycogen in the hepatopancreas increased significantly at the highest ammonia-N concentration. Concentration of oxyhemocyanin, acylglycerol, and cholesterol in hemolymph, and lactate in muscle decreased significantly in the group exposed to the highest ammonia levels. The changes observed in hemolymph and tissue metabolic fuels suggest a reduced use of carbohydrate through anaerobic metabolism and an increase in the use of lipids to satisfy the metabolic demand.

Food aversion learning induced by lithium chloride in the crayfish Procambarus clarkii.

The ability of the crayfish Procambarus clarkii to develop a food aversion learning was examined. For this purpose, in crayfishes fed with trout, a novel food (chicken) was associated in a single trial with a well-known toxic substance, lithium chloride (LiCl), and food intake was recorded for 10 days afterward using a two-choice protocol. All during the experimental period, a significant effect of treatment was observed that accounted for the lower chicken intake in the LiCl-treated group, compared with the control groups (NaCl-treated group and puncture group). In contrast, LiCl-treated animals had an increased trout intake during the entire period. In addition, it was observed that total food intake was similar among the three groups. These results indicate that the crayfish may develop a food aversion learning when illness is induced by LiCl after the animal accepts and eats a novel food.

Catecholamines in paraganglia associated with the hepatic branch of the vagus nerve: effects of 6-hydroxydopamine and reserpine.

Paraganglia are clusters of cells containing catecholamines (CA), mainly norepinephrine (NE) and dopamine (DA). The presence of epinephrine (E), on the other hand, has only been determined by indirect methods in retroperitoneal paraganglia of newborn and aged rats. Because their location, paraganglia associated with the hepatic branch of the vagus nerve may be a possible source of CA for the liver. The main purposes of the present study were to determine CA levels and whether E can be found in the omentum minus which includes paraganglia associated with the hepatic branch of the vagus nerve, and then to study the effects of 6-hydroxydopamine and reserpine on their CA content. Twenty-four female Wistar rats were randomly ascribed to three groups receiving two intraperitoneal injections of either 6-hydroxydopamine, reserpine or saline. Twenty-four hours after the last administration the rats were anesthetized and a portion of the omentum minus was obtained. Left adrenal medulla and a liver fragment were also collected as controls. The samples were processed to be analyzed by high performance liquid chromatography and catecholamine histofluorescence. The results confirm previous reports about the presence of considerable amounts of norepinephrine and dopamine in paraganglia. Norepinephrine and dopamine in the omentum like the adrenal medulla were significantly depleted by reserpine but not by 6-hydroxydopamine treatment, suggesting that some other sources in addition to sympathetic terminals are responsible for CA in the omentum. On the contrary, both drugs reduced liver NE, consistent with the localization of this amine mainly to hepatic sympathetic terminals. Histofluorescence of the omentum revealed 2-4 paraganglia per tissue fragment. Paraganglia associated with the hepatic branch of the vagus nerve contain also E. The presence of perihepatic sources of extra-adrenal CA, and more specifically E, could be of physiological significance.

Antiemetic inhibits conditioned taste aversion, but not the hypophagia induced by epinephrine.

It has been shown that relatively high doses of epinephrine (E) injected intraperitoneally (IP) produce hypophagia and conditioned taste aversion (CTA) in rats. We examined the possibility that E effects involve malaise. For this purpose, changes in saccharin preference induced by E injected IP (100 microg/kg) were determined after a previous administration of trimethobenzamide (TMB, 5 mg/kg), an antiemetic agent. E alone decreased saccharin preference by 54% (p < 0.01), but only by 16% (not significant) in the presence of TMB. In contrast, the injection of 75 or 100 microg/kg E reduced food intake by 50 and 85%, respectively (p < 0.01), regardless of previous injection of TMB. In conclusion, the results suggest that E-induced malaise is not the direct cause of the hypophagia it elicits.

Glucoprivation attenuates the hypophagia induced by epinephrine in mice.

It is well known that relatively high doses of epinephrine (E) injected intraperitoneally (IP) produce hypophagia, possibly by an action on liver metabolism. The purpose of the present experiment was to find out if lipoprivation with 2-mercaptoacetate (MA, 800 mumol/kg, IP) or glucoprivation with either 2-deoxy-D-glucose (2DG, 500 mg/kg, IP) or 2,5-anhydro-D-mannitol (2,5-AM, 400 mg/kg, IP) were able to modify the anorectic effect of E (300 micrograms/kg). At the onset of the dark period, mice received a first injection of saline (S) or one of the metabolic blockers mentioned above and, 30 min later, a second injection of S or E; then 30-min food intake was measured. E alone decreased feeding by 80% (p < 0.05); this effect was nearly the same when MA was previously injected. In contrast, in the presence of 2DG or 2,5-AM, E reduced food intake only by 22% and 24%, respectively (not significant). Attenuation of E-induced hypophagia by these blockers suggests the participation of glucose utilization pathways. Because it has been shown that 2,5-AM acts specifically on the liver, we could additionally suggest that E reduces feeding by an action on glucose hepatic metabolism.

Effect of lipectomy and long-term dexamethasone on visceral fat and metabolic variables in rats.

Intraperitoneal (IP) fat accumulation in humans is a risk factor for a number of diseases. We tried to increase this particular adipose mass in rats by long-term administration of low-dose dexamethasone (Dex) and/or elimination of other fat depots. Male adult Wistar rats were lipectomized (Lip) or sham-operated (Sh). Bilateral lipectomy of retroperitoneal and inguinal fat pads was performed under anesthesia with Na pentobarbital 40 mg/kg supplemented with ether. After 8 days, half the animals of each group received Dex in their drinking water (0.1 microgram/mL) while the other half received water (W), for a total of four groups: Sh-W, Lip-W, Sh-Dex, and Lip-Dex. Body weight (BW) and food and water intake were measured throughout the treatment period. A glucose tolerance test was performed 34 days after starting Dex treatment, and then rats were killed, fat depots were weighed, and plasma and liver were obtained for metabolic determinations. Dex rats ate the same amount of food as W controls, but gained significantly less weight (2.02 +/- 0.18 v 3.82 +/- 0.10 g/d, P < .01). Mean daily W intake was approximately 40 mL/d in all groups, which means that Dex rats ingested approximately 4 micrograms/d Dex. Average glycemic values during the 180-minute glucose tolerance test were as follows: Sh-W, 162 +/- 13; Lip-W, 166 +/- 7; Sh-Dex, 118 +/- 6; and Lip-Dex, 229 +/- 27 mg/dL. These values show that glucose tolerance was improved by Dex treatment alone, but was impaired in Lip-Dex animals. The same trend was evident for the relative weights (percent of BW) of two intact adipose depots: IP and epididymal (EPI) (Sh-W, 2.08 +/- 0.13 and 1.35 +/- 0.11, respectively; Lip-W, 1.67 +/- 0.15 and 1.17 +/- 0.11; Sh-Dex, 1.66 +/- 0.10 and 1.28 +/- 0.07; Lip-Dex, 2.41 +/- 0.11 and 1.53 +/- 0.09). Average glycemia for all rats was significantly correlated with IP (r = .55, P < .01) but not with EPI; moreover it was correlated in the Sh-W control group (r = .81, P < .05), suggesting a normal relation between these variables. Liver triglycerides (LTG), which were elevated in Dex rats, were also correlated with IP (r = .51, P < .02 for all rats and r = .82, P < .05 for Sh-W rats). The results show that long-term administration of low-dose Dex has some different effects in normal versus Lip rats concerning mainly the IP fat depot, the relative mass of which seems to significantly affect glucose tolerance.

Metabolic and hormone-related responses to caffeine in rats.

The time course of effects of caffeine on plasma glucose and non-esterified fatty acids (NEFA) were measured and related to various hormonal responses associated with substrate mobilization and utilization. Participation of the sympatho-adrenal system (SAS) in the metabolic and hormonal actions of caffeine was also investigated by the use of ganglionic blockade. Following 50 mg kg-1 i.p. injections of caffeine in rats, plasma glucose increased 25% and NEFA 40%, and these actions were parallelled by an elevation of plasma insulin, ACTH and corticosterone, without changes in glucagon. It is suggested that the insulin response is related to the plasma glucose increase and possibly also to an action of cAMP. When caffeine was injected in rats previously treated with the ganglionic blocker, hexamethonium, none of the responses mentioned above were modified. These results show that the glucose and NEFA responses are independent of glucagon secretion and are due not only to SAS activation but also to other mechanisms such as the increased ACTH and corticosterone secretion. It is also suggested that the mobilization of substrates by caffeine is mediated, through these various mechanisms, by the activation of cAMP and by phosphodiesterase inhibition.

Dissociation of anorectic and affective responses to epinephrine and glucose in rats.

The purpose of this study was to determine whether the anorexia following epinephrine and glucose IP injections is due to the activation of mechanisms of satiety. Epinephrine (100 micrograms.kg-1) and glucose (4 g.kg-1) were injected IP in rats. In control sessions for epinephrine test, rats received IP saline, and IM epinephrine. In control sessions for the glucose test, rats received IP NaCl, isoosmotic to the glucose solution. Food intake or taste reactivity to a sucrose solution was recorded after these treatments. Epinephrine and glucose decreased food intake by 75% (p < 0.001), and 49% (p < 0.01), compared to their controls. No change of taste reactivity responses was observed with any of these treatments. Twelve-hour fasting did not modify the general taste reactivity responses when compared to the responses evoked in rats fed ad lib. These results might be explained by the fact that anorexia could be obtained by a suppression of hunger without the activation of the mechanisms of satiety. This in turn would imply a possible dissociation between the signals and physiological pathways normally involved in hunger and satiety.

[Effect of adrenaline on food consumption and C-fos expression in the central nervous system]. / El efecto de la adrenalina sobre el consumo de alimento y la expresión de C-fos en el sistema nervioso central.

Intraperitoneal (IP) or intraportal epinephrine (E) administration produces strong hypophagia in rats whereas intravenous or intramuscular (IM) injection does not. These results suggest that E acts on liver to control food intake through vagal afferents to the brain. In the present work, immediate-early gene c-fos expression was used as an index of neuronal activity, comparing the respective effects of IP and IM E on food intake and on activation of brain areas that receive vagal information. Male Wistar rats were IP or IM injected with saline or E (100 micrograms/kg). In a first experiment, food intake was measured. In a second experiment, c-fos expression in different brain areas was assessed immunohistochemically. IP E administration reduced food intake by 75% (p < 0.01) whereas IM E had no effect. C-fos expression results showed that those solitary tract nucleus/area postrema regions receiving gastrointestinal and hepatic vagal afferents were specifically activated by IP E administration. These results support the possibility that E decreases food intake through a hepatic action involving vagal sensory neurons. However, higher integration levels of vagal information, such as lateral parabrachial nucleus or paraventricular nucleus, do not seem to be implicated in IP E effect on food intake.

The role of conditioned taste aversion in the hypophagia induced by intraperitoneal epinephrine and glucose.

It has been repeatedly shown that relatively high doses of epinephrine (E) and glucose (G) injected intraperitoneally (ip) produce hypophagia in fasted rats. In the present work we used a conditioned taste aversion (CTA) paradigm in order to test whether this effect could be due to "malaise." We determined the effect on food intake and saccharin preference with the following treatments: (a) E ip 100 and 250 micrograms/kg; (b) E ip 250 micrograms/kg with or without previous alpha 1 plus beta adrenergic blockade; (c) G ip 3.5 and 4 g/kg. Both doses of E significantly reduced food intake more than 75% but only the high dose produced a significant (50%) reduction in saccharin preference. Blockade of alpha 1 and beta adrenergic receptors completely suppressed the E-induced hypophagia but attenuated only slightly the taste aversivon effect. Both doses of G decreased food intake but only the high dose reduced saccharin preference; part of these effects would appear to be due to the high osmolarity of the solution. The present results indicate that E and G may induce CTA in our experimental conditions. However, their hypophagic and aversive effects seem to be elicited by different mechanisms.

The effect of caffeine on food intake in rats: involvement of corticotropin-releasing factor and the sympatho-adrenal system.

The involvement of CRH and the sympatho-adrenal system in the effects of caffeine on food intake and body weight gain has been investigated in rats. Food intake and body weight gain were measured in male rats after the treatment with caffeine in combination with either an injection of the CRH antagonist alpha-helical CRH(9-41), a surgical adrenal demedullation (medullectomy), or a ganglionic blockade. Alpha-helical CRH(9-41) was injected in the lateral ventricle of the brain and hexamethonium was used to chemically block the ganglionic transmission. From 4 to 24 h following a caffeine injection, spontaneous food intake, which was cumulated from the time caffeine was injected, was significantly (p < 0.01) lower in caffeine- than in saline-treated rats. In food-deprived rats, the anorectic effect of caffeine was biphasic, being significant at 0.5 and 1 h after the caffeine administration, then vanishing for 3 h, and becoming significant again 6 h after the caffeine administration. In both the spontaneously fed and food-deprived rats, caffeine reduced the rate of weight gain, which was measured at the end of a 12- or a 24-h period following the caffeine injection. A significant (p < 0.05) interaction effect of caffeine and alpha-helical-CRH(9-41) was found on the cumulative food intake at 1, 6, and 8 h, and on the amount of food eaten between the 4-6-h interval following the injection of caffeine; the effects of caffeine on food intake and body weight gain seem largely prevented by the use of a CRH antagonist.(ABSTRACT TRUNCATED AT 250 WORDS)