The biochemical composition of the pyloric caeca and its relation to DNA levels in the female starfishAsterias rubens during the reproductive cycle

Summary The biochemical composition of the pyloric caeca of femaleAsterias rubens was studied during the annual reproductive cycle. The amounts of nucleic acids, DNA and RNA were measured in addition to the amounts of lipids, glycogen, other reducing carbohydrates, free amino acids and proteins.A positive correlation of the pyloric caeca index to the biochemical constituents at the cellular level was established. The total DNA content of the pyloric caeca was found to increase with the pyloric caeca index.The relation of the biochemical composition of the pyloric caeca to the size changes of the organ is discussed. The hypothesis that the changes in size of the organ are due to changes both in the number and size of the cells is confirmed. The biochemical composition of the pyloric caeca and its relation to the reproductive cycle is discussed.Abbreviations GI gonad index - CPI pyloric caeca index     

Insectivore to frugivore: ontogenetic changes in gut morphology and digestive enzyme activity in the characid fish Brycon guatemalensis from Costa Rican rain forest streams

Brycon guatemalensis , a Neotropical characid fish, consumes an entirely terrestrial diet, shifting from eating insects as juveniles to fruits and leaves as adults. Juvenile and larger‐sized fish collected in the Rio Puerto Viejo at the La Selva Biological Station in Costa Rica were studied to test the hypotheses that, with ontogeny, (1) relative gut length increases, (2) pyloric caeca arrangement and number remain unchanged and (3) pepsin, trypsin and lipase activities decrease, while α‐amylase activity increases. These hypotheses were mainly supported in that larger fish had longer guts, unchanged pyloric caeca arrangement but fewer caeca, and, at both environmental and standard temperatures for the enzyme assays, lower pepsin and trypsin activities but higher α‐amylase activities than the juveniles. Only lipase, among the digestive enzymes, exhibited the unexpected outcome of either not differing significantly in activity (per g of tissue) between juveniles and larger fish or being significantly higher (per mg of protein) in larger fish. The overall results support the view that B. guatemalensis is specialized morphologically and biochemically to function first as a carnivore and then as a herbivore during its life history.     

An autoradiographic investigation of the proliferative activity of pyloric caeca and gondas of Asterias rubens

The proliferative activity of the pyloric caeca of Asterias rubens was investigated. Autoradiographic experiments using intracoelomically injected (methyl-³H)-thymidine were performed throughout the year and incorporation into pyloric caeca and into gonads was studied. Tritiated thymidine was found to be incorporated mainly in the coelomic lining of both organs. Cell divisions in the coelomic lining may be necessary for the growth of these organs, for the production of coelomocytes or, in the case of the pyloric caeca, for growth of the digestive epithelium. Proliferative activity of the digestive epithelium of the pyloric caeca was only observed in the median duct. It is hypothesized that new cells, arising from mitosis, grow from the median duct to the side lobes and differentiate into storage cells, for example. The existence of a mitosis-inducing or mitosis-stimulating substance is discussed. In the ovaries follicle cells were found to incorporate (methyl-³H)-thymidine; in the testis, proliferation of the germinal epithelium occurred simultaneously in all spermatogenic columns. First, the spermatogonia and then later the spermatocytes became labeled. Absorption of substances from the coelomic fluid is discussed.     

Lipolytic enzymes of the digestive organs of the crown-of-thorns starfish (Acanthaster planci): comparison of the stomach and pyloric caeca

1. Stomach and pyloric caeca homogenates from the crown-of-thorns starfish hydrolysed p-nitrophenyl esters, alpha-naphthyl esters, cholesteryl oleate and tributyrin. The pyloric caeca contained the highest activities. 2. The p-nitrophenyl acetate hydrolytic activity eluted at 0.23 M NaCl on ion exchange chromatography while the p-nitrophenyl palmitate hydrolytic activity eluted between 0.2 and 1.0 M NaCl. 3. Polyacrylamide gel zymograms for alpha-naphthyl acetate hydrolytic activity revealed one major band and several minor bands of activity for both tissues. 4. Isoelectric focusing zymograms revealed one major band with a pI = 4.2 for both tissues, with an additional band at pI = 3.5 for pyloric caeca. 5. The pyloric caeca contained twice as much lipid as the stomach. Lipid extracts contained mixtures of steroids and steroid-esters; a cholesterol-like sterol was tentatively identified.     

Nitric oxide (NO) synthase immunoreactivity in the starfish Marthasterias glacialis

The neuroendocrine system of the starfish Marthasterias glacialis was investigated immunocytochemically using antisera specific for rat neuronal, bovine aortic endothelial, and mouse macrophage, nitric oxide (NO) synthases. Immunoreactivity was detected only with the antibodies specific for the neural enzyme, in the ectoneural and hyponeural tissues of the radial nerve cords and in the basiepithelial plexus and endocrine cells of the digestive tract. The pyloric stomach showed more immunoreactive structures than the other digestive organs, with the rectal caeca showing the least activity. Immunoreactive endocrine cells were located in the cardiac and pyloric stomachs and in the pyloric caeca. Co-localization of the enzyme immunoreactivity, and the staining for NADPH-diaphorase, demonstrate the presence of NO synthase in echinoderms. These results provide further evidence that NO is a neuronal messenger of early phylogenetic origin which has been conserved throughout evolution.     

Adipokinetic hormones control amylase activity in the cockroach (Periplaneta americana) gut

This study examined the biochemical characteristics of α‐amylase and hormonal (adipokinetic hormone: AKH) stimulation of α‐amylase activity in the cockroach (Periplaneta americana) midgut. We applied two AKHs in vivo and in vitro, then measured resultant amylase activity and gene expression, as well as the expression of AKH receptor (AKHR). The results revealed that optimal amylase activity is characterized by the following: pH: 5.7, temperature: 38.4 °C, K_m (Michaelis–Menten constant): 2.54 mg starch/mL, and V_max (maximum reaction velocity): 0.185 μmol maltose/mL/min. In vivo application of AKHs resulted in significant increase of amylase activity: by two‐fold in the gastric caeca and 4–7 fold in the rest of the midgut. In vitro experiments supported results seen in vivo: a 24‐h incubation with the hormones resulted in the increase of amylase activity by 1.4 times in the caeca and 4–9 times in the midgut. Further, gene expression analyses reveal that AKHR is expressed in both the caeca and the rest of the midgut, although expression levels in the former were 23 times higher than levels in the latter. A similar pattern was found for the amylase (AMY) gene. Hormonal treatment did not affect the expression of either gene. This study is the first to provide evidence indicating direct AKH stimulation of digestive enzyme activity in the insect midgut, supported by specific AKHR gene expression in this organ.     

Retrograde peristalsis; a possible mechanism for filling the pyloric caeca?

A marked, relatively rapid retrograde peristaltic activity was observed in the pyloric region of Atlantic halibut Hippoglossus hippoglossus in vitro and in vivo. This could provide a mechanism for filling the pyloric caeca, and the retrograde contraction could also be involved in mixing the chyme with digestive secretions in the region of the pyloric caeca.     

Physiological correlates of growth and condition in the yellow perch (Perca flavescens)

This study on yellow perch (Perca flavescens) examines a series of enzymatic markers and the relative weights of pyloric caeca and visceral lipids, their response to changes in feeding regime and their potential use to infer recent changes in growth rate and fish condition. Fish were exposed to four different feeding regimes for 12 weeks resulting in specific growth rates ranging from 0.3% to 3.5% (%/day). Growth and condition responded rapidly to changes in ration and the weight of pyloric caeca and visceral lipids reflected increased feed intake. Growth rate was correlated with muscle citrate synthase and caecal nucleoside-diphosphate kinase activities, whereas condition was correlated with muscle citrate synthase and lactate dehydrogenase activities and with caecal glucose-6-phosphate dehydrogenase activity. Results showed that enzyme activities and biometric parameters responded rapidly to increased feed intake, but the response was slower when food intake decreased. Plateaus were attained for both condition and visceral lipid index, but the relative weight of pyloric caeca continued to increase throughout the experimental period. Results from this study could, in principle, be used to infer recent growth and energy status in wild yellow perch and thus provide an indicator of food availability in their environment.     

Triloborhynchus psilastericola n. sp., a parasitic turbellarian (Fam. Pterastericolidae) from the starfish Psilaster andromeda (Müller and Troschel)

The turbellarian Triloborhynchus psilastericola n.sp. (family Pterastericolidae) occurs in the starfish Psilaster andromeda. On approaching maturation it moves from the host's coelom into the glandular pockets of the pyloric caeca with simultaneous loss of almost the entire ciliation. Mature turbellarians feed on the tissue of the pyloric caeca.     

The annual pyloric cycle of Asterias rubens L. (echinodermata: Asteroidea)

Observations of biochemical changes in the pyloric caeca of Asterias rubens L. in different nutritional states and at different stages of the reproductive cycle show the existence of an annual cycle in the pyloric caeca. This pyloric cycle is closely related to that of the gonads. During the gonadal rest (May to September, when the gonads are barely visible) the pyloric caeca grow and store glycogen. During gonadal activity (October to April), they progressively autolyse and first accumulate, then liberate, lipid reserves. This winter pyloric autolysis provides the gonads with the structural material necessary for their annual growth. The summer pyloric growth must be interpreted as the accumulation of the structural material. Under normal conditions, during gonadal rest, the pyloric glycogen reserves ensure the survival of the animal. Pyloric lipid reserves appear only at the beginning of gonadal activity. Their synthesis is intimately related to the start of the gonadal activity of which they are the principal energy source. We believe that the pyloric cycle is under control of hormones that could be transported through the haemal system.The pyloric caeca are the only reserve organs of starfish. Neither the other digestive organs nor the body wall accumulate energy reserves. During starvation of medium duration (2 to 4 weeks) the pyloric caeca suffice to ensure the survival of the animal by liberating their reserve material and simultaneously starting to autolyse. During prolonged starvation, general autolysis occurs and the size of the starfish diminishes.     

The Effect of 17 β-Estradiol and Estrone on Intermediary Metabolism of the Pyloric Caeca of the Asteroid Luidia clathrata (Say) Maintained under Different Nutritional Regimes

Luidia clathrata were fed either a high or low level of food and injected with estradiol, estrone, or seawater for 16 days. In individuals maintained at a low level of food, the weight of the pyloric caeca decreased in all individuals, regardless of hormone injections. Activities of all enzymes in these individuals were lower than in individuals maintained at a high level of food. In the individuals maintained at a low level of food, the activity of GPDH was higher in the pyloric caeca of individuals injected with estradiol and estrone than in those receiving seawater injections; the activity of 6-PGDH decreased in those injected with estrone. In individuals maintained at a high level of food, the activities of glucose-6-phosphate dehydrogenase (G–6–PDH) and 6-phosphogluconate dehydrogenase (6–PGDH) were higher in individuals injected with estradiol than in those receiving seawater injections. Activities of glycerol phosphate dehydrogenase (GPDH) and pyruvate kinase (PK) were not affected. A slight decrease (P<0.1) in the activity of 6–PGDH was seen with injection of estrone. These results suggest that steroids have a role in regulating cellular metabolism in the pyloric caeca, and that nutritional condition influences the response of the pyloric caeca to steroids. In addition, these results indicate that the nutritional condition of the individual directly influences enzyme activities.     

Allocation of nutrients during the reproductive cycle of Ophidiaster ophidianus (Echinodermata: Asteroidea)

The reproductive cycle of Ophidiaster ophidianus (strictly protected status) from São Miguel Island, in the Azorean Archipelago was studied. The reproductive strategy; the energy allocation of each sex during the reproductive cycle and the nutritional condition of the population were analyzed. Gonadal index (GI) showed a clear seasonal pattern with spawning between August and October but histological examination revealed that gamete release can occur throughout the entire year. The pyloric caeca index (PCI) showed little annual variation but with an inverse relationship with the GI. Allocation of energy to the gonads and to the pyloric caeca reflected the seasonal reproductive strategy of this species. Individuals were able to simultaneously develop gonads, pyloric caeca, and quickly regenerate lost arms. There was a major expenditure of energy by females compared to males but, sexual size dimorphism was not observed. The reproductive pattern observed in O. ophidianus combining rich food availability and seawater temperatures characteristic of a temperate zone may be the key to the success of this species in the Azorean oceanic Island.     

The effect of adipokinetic hormones on the activity of digestive enzymes

The role of the adipokinetic hormone (AKH) in the control of protease, amylase and lipase activities is examined using the cockroach Periplaneta americana and the fruit fly Drosophila melanogaster as model species. The effects of Peram‐CAH‐I and ‐II on the activity of cockroach digestive enzymes in the gastric caeca and midgut are measured both in vivo and in vitro. The results show the activity of proteases, amylases and lipases in both parts of the gut: amylase activity is higher in the gastric caeca than in the midgut; lipase activity presents the opposite trend; and protease activity is similar in both organs. The applied hormones stimulate the activity of all digestive enzymes, although this stimulation is not uniform; AKHs affect enzymes selectively, and in some cases unequally, in the gastric caeca and midgut. No substantial differences between Peram‐CAH‐I and ‐II stimulation are recorded. The in vitro results demonstrate that AKH stimulates digestive enzyme activity directly. In agreement with the cockroach results, enzymatic activity in D. melanogaster larvae producing nonfunctional AKH is lower than that in the larvae with ectopically expressed Akh gene, where enzyme activity reaches or even exceeds that of the controls. Overall, the results demonstrate the active role of AKHs in the stimulation of digestive enzyme activity in insects.     

Phenotypic flexibility of digestive system in Atlantic cod (Gadus morhua)

This study examined the restoration of the digestive capacity of Atlantic cod (Gadus morhua Linnaeus) following a long period of food deprivation. Fifty cod (48 cm, 1 kg) were food-deprived for 68 days and then fed in excess with capelin (Mallotus villosus Müller) on alternate days. Ten fish were sampled after 0, 2, 6, 14 and 28 days and the mass of the pyloric caeca, intestine and carcass determined. Two metabolic enzymes (cytochrome c oxidase and citrate synthase) were assayed in white muscle, pyloric caeca and intestine, and trypsin activity was measured in the pyloric caeca. A delay of 14 days was required before body mass started to increase markedly, whereas most of the increase in mass of both the pyloric caeca and intestine relative to fish length occurred earlier in the experiment. By day 14, the activities of trypsin and citrate synthase in the pyloric caeca as well as citrate synthase in the intestine had reached maxima. The growth of the digestive tissues and restoration of their metabolic capacities thus occur early upon refeeding and are likely required for recovery growth to take place. The phenotypic flexibility of the cod digestive system is therefore remarkable: increases in trypsin activity and size of pyloric caeca resulted in a combined 29-fold increase in digestive capacity of the fish during the refeeding period. Our study suggests that Atlantic cod are able to cope with marked fluctuations in food availability in their environment by making a rapid adjustment of their digestive capacity as soon as food availability increases.     

The effects of various diets on chitinase and ß-glucosidase activities and the condition of cod, Gadus morhua (L.)

Groups of cod, Gadus morhua (L.), were fed exclusively on fish, crustaceans, or crustacean shells for a period of 3 weeks. Chitinase and ß-glucosidase activities were measured in enzyme extracts of stomach contents, stomach tissue, pyloric caeca, intestinal contents, and intestine tissue, and compared to the enzyme activities of control fish starved over the same period. Fulton's condition factor K , liver lipid content and liver water content were determined to estimate the effects of the diets on the condition of cod.
general, the highest chitinase activities were measured in samples of cod that had been fed on whole crustaceans. In this feeding group, there was also a remarkable increase of activity by a factor of four to eight in the pyloric caeca, compared to the group fed on fish and the control group, respectively.
Measurements of ß-glucosidase activity revealed no similar dependence on food quality, ß-glucosidase occurrence seemed to be mainly restricted to the pyloric caeca and the intestine of cod.     

Histochemical observations on the pyloric caeca of Asterias rubens (Echinodermata,asteroidea) in relation to the reproductive cycle

The pyloric caeca of the starfish Asterias rubens were investigated histochemically during the reproductive cycle. The median duct and the side lobes reacted differently. The median duct reacted positively for acid phosphatases and glucose-6-phosphate dehydrogenase, whereas the side lobes reacted positively for alkaline phosphatases, neutral lipids, and fatty acids. In the transition zone between the median duct and the side lobes, the reaction for alkaline phosphatases and neutral lipids increased toward the side lobes. The function of the enzymes and the histochemical results are discussed in relation to the function of the pyloric caeca and to the reproductive cycle.     

Level,content, and caloric equivalents of the lipid,carbohydrate, and protein in the body components of Luidia clathrata (echinodermata: asteroidea: platyasterida) in Tampa bay

The body components of the primitive platyasterid starfish, Luidia clathrata (Say) have been characterized according to season, reproductive, and nutritional condition. The animal has been ‘reconstructed’ in terms of the wet and dry weights, the weight of organic matter, and the caloric equivalents of the body components in early summer (when there are no gonads and the pyloric caeca are small), in autumn (just prior to initiation of gonadal development and at maximal caeca size), and in early spring (at maximal gonad size). The body wall is always the most significant component in terms of wet and dry weights. In terms of organic matter and calories, the pyloric caeca are the most significant compartment in the fall and, together with the gonads, in the spring. On starvation for one month, the decrease in the size of the pyloric caeca is calculated to have produced 2.048 kcal. The energy requirement over the period, calculated from Q_o2 values, is 2.112 kcal. There was no change in biochemical composition of the pyloric caeca with starvation, suggesting that energy production came about through cell destruction rather than preferential utilization of cellular nutrient stores. Lack of extreme change in the composition of the pyloric caeca with size through the year also suggests that energy deposition in the organ is primarily by change in cell number and not by cell size.     

The response of digestive proteases to abrupt salinity decrease in the euryhaline sparid Sparus aurata L

The response of the digestive proteases to abrupt salinity change was studied in juvenile gilthead sea bream (Sparus aurata) for 15 days after transfer from 33 per thousand to 21 per thousand. Salinity decrease affected significantly neither the activity of total acid proteases in stomach, nor the activities of total alkaline proteases and major serine proteases--trypsin and chymotrypsin--in the alkaline part of the intestine. The activity of the major proteases was significantly different between the alkaline segments of the intestine, with the posterior intestine presenting the highest activities followed by the pyloric caeca. This distribution pattern remained unaffected by salinity decrease. Notably, salinity change led to significant alterations in elastase and carboxypeptidase activity. The changes were more prominent in the upper part of the intestine (pyloric caeca and anterior intestine) than in the posterior intestine. In pyloric caeca significant alteration of carboxypeptidase A and B activities was observed, elastase changes were confined to anterior intestine together with alterations in carboxypeptidase B activity, while in posterior intestine the changes were restricted to carboxypeptidase A activity. The results are discussed in relation to the osmoregulatory action of the intestinal segments and dietary protein digestion.     

Carbohydrase activity in the digestive system of some teleost fishes.

The activity of carbohydrases in Puntius sophore (Ham.), Channa gachua (Ham.) and Cirrhinus mrigala (Ham.) has been studied. The carbohydrases have been found in the stomach, intestinal bulb, intestine, pyloric caeca and the hepato-pancreas. The hepatopancreas is the main site of production of these enzymes and it is in this organ and the intestine that their activity is highest. Their pH optimum lies between 5.4 and 6.4. The enzyme equipment in the teleost is adapted to their respective food and feeding habits both qualitatively and quantitatively. In Puntius (omnivorous) and Cirrhinus (herbivorous) all three carbohydrases, namely amylase (EC 3.2.1.1.), sucrase (EC 3.2.1.26.) and raffinase, while in Channa (carnivorous) only amylase and sucrase have been found to be active. In Cirrhinus mrigala, which is predominantly a herbivorous species, the concentration of carbohydrases is higher than those in the other two fishes.     

Morphology of the digestive tract of the Whitemouth croaker Micropogonias furnieri (Desmarest, 1823) (Perciformes: Sciaenidae)

Studies have shown the feeding diversity of teleostean fish, which is conditioned by environmental characteristics or the biology of the different species. Analysis on the morphology of the digestive system (DS) of fish made it possible for researchers to know the food regimen of several species at different stages of life. On the other hand, it is known that food habits may lead to morphological changes in the DS the same way that different food habits may be imposed by morphological limitations of this system. Among the species of greater commercial importance in Brazil, Micropogonias furnieri is highly representative of fish in south‐eastern Brazil. Therefore, the objective of this study was to analyse the morphology of the DS of M. furnieri. Results on the morphology of the DS observed in this study were similar to the patterns determined for the morphology of teleostean fish. In this study, it was observed that the DS of the Whitemouth croaker is directly related with the shape of the peritoneal cavity; these fish showed oesophagus, stomach, pyloric caeca and intestines. The pyloric caeca showed morphological adaptations in relation to sexual maturation, with well‐developed caeca found in sexually mature animals.