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Abstract The study was carried out to evaluate wedge wire screen as a potential tool for collecting fish faeces from the tank outlet water. Apparent digestibility (AD) estimates of carbon (AD C), nitrogen (AD N), sulphur (AD S), organic material (AD ORG) and individual amino acids obtained by the wire screen were compared with stripping. Three diets, with fishmeal, soybean meal (SBM) or rapeseed meal (RSM) as main protein source were extruded and fed to triplicate groups of 120-g rainbow trout (Oncorhynchus mykiss). Faeces were obtained by gentle stripping from the distal abdomen, and by collection from a wedge wire screen 15, 30, 60, 120 and 240 min post feeding. AD ORG , AD N and AD S obtained by collection on the screen were significantly (P <.05) higher than those obtained by stripping, except for some values for AD C. Differences in AD between stripping and faeces collected with the wire screen were lower than reported for other methods of collection from water media. Most of the increase in AD of nutrients for wedge wire screen occurred during the first 15 min after defecation. The AD ORG , AD C , AD N and AD S were lowest in the RSM diet. The AD of amino acids followed the same pattern as AD N. AD S in the rapeseed diet was particularly low (54.7%), compared to the fishmeal (72.4%) and SBM diet (70.1%). In addition to AD of cysteine, low digestibility of glucosinolates, its degradation products and other sulfonated components are likely reasons for the low AD S. Faeces from the RSM based diet had higher dry matter (DM) concentration than the SBM based diet. Faecal DM recovery for SBM was lowest among the diets for both collection methods. The low DM recovery for the SBM faeces did not affect AD estimates of faeces at the different time intervals, possibly due to simultaneous leakage of nutrients and indigestible marker. In conclusion, the use of wedge wire screen for collection of faeces represents an interesting supplement to stripping. The AD estimates had low random variation and nutrient leaching during faeces collection were dependent on dietary composition. Highlights • The digestibility estimates from the wire screen had low variation within time points and dietary treatments. • Digestibility results obtained for faeces collected from water should not be used as indicator for fecal nutrient loss • Feed composition and characteristics play a significant role in nutrient loss from faeces. [ABSTRACT FROM AUTHOR]

Metabolic rates vary widely within species, but little is known about how variation in the 'floor' [i.e. standard metabolic rate (SMR) in ectotherms] and 'ceiling' [maximum metabolic rate (MMR)] for an individual's aerobic scope (AS) are linked with digestive and locomotor function. Any links among metabolic traits and aspects of physiological performance may also be modulated by fluctuations in food availability. This study followed changes in SMR, MMR, and digestive and locomotor capacity in southern catfish (Silurus meridionalis) throughout 15 days of food deprivation and 15 days of refeeding. Individuals downregulated SMR during food deprivation and showed only a 10% body mass decrease during this time. Whereas critical swim speed (Ucrit) was robust to food deprivation, digestive function decreased after fasting with a reduced peak oxygen uptake during specific dynamic action (SDA) and prolonged SDA duration. During refeeding, individuals displayed rapid growth and digestive function recovered to pre-fasting levels. However, refed fish showed a lower Ucrit than would be expected for their increased body length and in comparison to measures at the start of the study. Reduced swimming ability may be a consequence of compensatory growth: growth rate was negatively correlated with changes in Ucrit during refeeding. Southern catfish downregulate digestive function to reduce energy expenditure during food deprivation, but regain digestive capacity during refeeding, potentially at the cost of decreased swimming performance. The plasticity of maintenance requirements suggests that SMR is a key fitness trait for in this ambush predator. Shifts in trait correlations with food availability suggest that the potential for correlated selection may depend on context. [ABSTRACT FROM AUTHOR]

Assessing the activity of digestive enzymes is a common procedure in many biological, physiological and nutritional studies. After reviewing the available literature on fish digestive tract maturation and enzymatic activity (pancreatic and intestinal enzymes) published between 1994 and 2017, authors detected some possible methodological and/or interpretative inconsistencies in this kind of studies, and concluded that special attention should be paid on: i ) the time of conservation of frozen samples prior their analysis, ii ) the proper purification of the brush border of enterocytes by a double centrifugation step (Crane et al., 1979) when authors want to evaluate the activity of intestinal brush border enzymes in order to avoid the overestimation, particularly of alkaline phosphatase (AP), because it is present in other tissues; iii ) the use of the proper reaction conditions at the normal range of values in terms of ions, temperature and intestinal alkalinity for the species of interest, and AP unit calculation. The implementation of these recommendations will promote the standardization of actual analytical procedures, as well as improve the reliability of comparative studies between different fish species or rearing procedures. [ABSTRACT FROM AUTHOR]

Elasmobranchii shows particular characteristics that make them susceptible to the impacts caused by fishing pressure, especially bottom trawling, which mostly affect rays, as they have demersal habits. Based on food availability and feeding ability, Polychaeta and Sipunculiforme are the bases of Brazilian electric ray Narcine brasiliensis diet. Morphologically, the digestion starts in the oesophagus with acid mucines action. The existence of acid and neutral mucines in the posterior intestine prevents lesions in the mucosa and favours the passage of faeces and shell residues. Thus, this study shows the interaction between their habitat, the organisms and their morphophysiology. [ABSTRACT FROM AUTHOR]

We describe the use of the fluorescent reporter compound CDr10b to label mid-intestinal structures in zebrafish larvae after simple immersion. CDr10b is deposited into the gut where it initially fills the lumen and is excreted. Using laser-mediated injury of the intestine, we show that CDr10b provides a useful readout of the integrity and repair of the epithelial cell barrier. In addition, CDr10b specifically labels the absorptive mid-intestine segment that is analogous to the mammalian small intestine. By perturbing retinoic acid signaling, which regulates the size of the mid-intestine segment, we show that CDr10b is a valuable tool to rapidly assess developmental malformations of the intestine in live animals. [ABSTRACT FROM AUTHOR]

An increase in salinity of the aquatic habitat can be an environmental stress factor for fresh aquatic organism, including fish. The present study investigated the impact of salinity and diet composition on digestibility and intestinal morphology of Nile tilapia. Triplicate groups of 35 fish weighing 35 g were fed a “Control” and “Test” diet and kept at salinity of 0‰ and 15‰ for 8 weeks. The diets were formulated using a different soybean meal content to create a contrast in alterations in intestinal morphology. Six fish per treatment were sampled for intestinal morphology analysis at the end of weeks 1, 4 and 8. The proximal, middle and distal intestine were processed for quantitative histology, in order to count the number of goblet cells and eosinophilic granulocytes; and to measure the thickness of lamina propria and sub-epithelia mucosa. The study showed that a salinity of 15‰ increased the nutrient digestibility; however this enhanced digestibility diminished over time. The intestinal morphology was influenced by both the salinity as well as the dietary soybean meal content. For soybean meal, the impact on morphology was largest in the proximal region of the intestine, whereas for salinity the largest impact was in the distal region. The negative effect of increased soybean meal on the lamina propria thickness was enhanced at a salinity of 15‰ and aggravated in time. Difference in nutrient digestibility induced by the salinity seemed not to be related to alterations in the intestinal morphology. [ABSTRACT FROM AUTHOR]

Carbohydrate utilization by fish varies with feeding habits and feed processing. Extruded aquafeeds have increased digestibility of carbohydrate sources resulting from the cooking and gelatinization of this nutrient, making it readily available for all aquatic species. This study aimed at determining the effects of extrusion cooking of feeds in the digestibility of different dietary carbohydrate sources for dourado. Isonitrogenous and isoenergetic diets (530 g kg−1 crude protein and 20.80 MJ kg−1 gross energy) containing either corn, toasted cassava flour, wheat flour, broken rice, cassava starch, or corn starch, in extruded or pelletized form and added of chromic oxide as inert marker, were fed to groups of juvenile dourado (73.97 ± 12.0 g; n = 40) stocked in 500-L polyethylene tanks. Extrusion cooking improved the digestibility of energy and/or starch of corn, cassava starch, wheat flour, rice and corn starch ( P < 0.05), but did not affect digestibility of toasted cassava flour ( P > 0.05). It is thus fair to infer that dourado, despite being a carnivorous species, is properly fit to digest wheat flour, rice and corn starch in natural form. [ABSTRACT FROM AUTHOR]

Fish remains are abundant and easily recognizable in many archaeological sites. The origin of such assemblages is generally poorly known. The ability to create small fish bone accumulations is restricted to a small number of predators (including raptors, carnivores and humans). In order to recognize the agents responsible for these concentrations in archaeological sites, we have investigated the taphonomy of the fish remains from otter spraints to determine the potential role of the Eurasian otter, Lutralutra, in the formation of fossil assemblages as a piscivorous mammal. Currently, few data are available to characterize the accumulation of otter remains in natural or archaeological sites. We analyzed a collection of 29 otter spraints from Bugeat (Corrèze) in central France. We identified the remains of eight fish species: brown trout (Salmotrutta, Salmonidae), perch (Percafluviatilis, Percidae), pumpkinseed (Lepomisgibbosus, Centrarchidae), and several Cyprinidae, rudd (Scardiniuserythrophthalmus), minnow (Phoxinusphoxinus), bullhead (Cottusgobio), and the indeterminate pair chub/dace (Squaliuscephalus/Leuciscusleuciscus). The species and the body parts present, along with the reconstruction of the fish weight and observations of any modifications, such as deformation, rounding and polishing, give us insight into otter prey categories and their transformation after digestion. Among the ca. 200 bones constituting the whole fish skeleton, only between six and 39 bones survived depending on the different species. Cyprinids display a high-moderate digestion grade and seem to be the taxon most affected by digestion (64.67%). Based on surface modifications, such as breakage and digestion traces, as well as bone element representation, we produced a set of criteria as a new tool to be applied to the fossil record. [ABSTRACT FROM AUTHOR]

This study aimed to investigate the optimal dietary vitamin B 12 requirement of fingerling blunt snout bream Megalobrama amblycephala based on the results of growth performance, intestinal digestive and absorptive capability and responses of the growth hormone (GH)-insulin-like growth factor-I (IGF-I) axis. Fish (initial body weight: 0.71 ± 0.03 g) were randomly assigned to six purified diets containing graded levels of vitamin B 12 (0, 0.028, 0.051, 0.12, 0.23 and 0.41 mg/kg) for 12 weeks. Both weight gain (WG) and specific growth rate increased significantly ( P < 0.05) as dietary vitamin B 12 levels increased from 0 to 0.051 mg/kg, then plateaued with further increasing levels. Feed conversion ratio decreased as dietary vitamin B 12 levels increased from 0 to 0.12 mg/kg, but no statistical difference was observed. Whole-body composition and intestinal brush border enzymes activities also exerted little difference within the vitamin B 12 range tested, whereas hepatic vitamin B 12 contents increased remarkably ( P < 0.01) with increasing dietary vitamin B 12 levels. Intestinal protease, amylase and lipase activities all increased significantly ( P < 0.05) as dietary vitamin B 12 levels increased from 0 to 0.12 mg/kg, but decreased remarkably ( P < 0.05) with further increasing vitamin B 12 levels. Similar results were also observed in hepatic IGF-I expressions. Furthermore, the mRNA expressions of GH in the brain decreased significantly ( P < 0.05) with increasing dietary vitamin B 12 levels. Based on the broken-line regression analysis of WG, the optimal dietary vitamin B 12 requirement of fingerling blunt snout bream was estimated to be 0.06 mg/kg. In addition, a supplementation of 0.12 mg/kg vitamin B 12 boosted intestinal digestive capability and hepatic IGF-I expression. Statement of relevance This study investigated the optimal dietary vitamin B 12 requirement of fingerling blunt snout bream based on growth performance. The underlying mechanisms were also investigated with emphasis on intestinal digestive and absorptive capability and modulation of the GH-IGF-I axis. Data obtained here might facilitate the development of low-cost formulated feed for aquatic species. [ABSTRACT FROM AUTHOR]

The serial replacement of fish meal (anchovetta) by canola meal (CM) (100, 200, 300 g kg − 1 as either solvent extracted (SE) CM or expeller extracted (EX) CM was undertaken to investigate the effects of increasing dietary CM levels on feed intake, growth, protein and energy retention, plasma biochemistry and the expression of a suite of hepatic genes in barramundi (Asian seabass; Lates calcarifer ) over an eight week feeding trial. An additional diet using lupin kernel meal (LM) to replace the fish meal was also included as a comparative reference. Eight iso-digestible nitrogenous (423 ± 29 g kg − 1 ) and iso-digestible energetic (14.6 ± 8 MJ kg − 1 DM) diets were formulated. Each diet was randomly allocated to triplicate groups of fish in seawater tanks (600 L), and each tank was stocked with 15 fish (53.4 ± 7.0 g). Fish were fed once daily (9:00–10:00) to apparent satiation, and uneaten feed was collected to determine feed consumption. The results showed that the survival, feed intake, growth, FCR, energy and protein retention of fish fed the diet containing SE CM were similar or even higher to those of fish fed the fish meal reference diet (FM) and the LM diet. Fish fed with the diet containing 300 g kg − 1 SE CM did not show any changes in biochemistry and gene expression in a suite of detoxification genes. However, the diet with 300 g kg − 1 EX CM depressed feed intake, growth performance and increased feed conversion ratio (FCR). Transcription of genes involving in fatty acid synthesis and the TCA cycle were not changed by different diets. The down regulation of gene expression in certain detoxification genes ( Lc CYP1A1 , Lc CYP3A , Lc CYP2N and Lc GST ) was observed in fish fed with the diet containing 300 g kg − 1 EX CM compared to the FM control diet and other experimental diets. In general, the SE CM can be used up to 300 g kg − 1 diet without negative performance effects or signs of clinical plasma biochemistry. By contrast the maximum acceptable level of the EX CM for barramundi was only 200 g kg − 1 . Higher inclusion level of the EX CM induced negative effects on growth performance, feed utilisation, plasma biochemistry and gene expression in relation to detoxification. Statement of relevance Previous research has demonstrated that canola meal is a potential plant protein source for fish meal replacement in diets for many fish species. The present work is designed to be the first to assess both two types of Australian canola meal in regard to different processing methods (solvent extraction and expeller extraction) in use for barramundi (Asian seabass) diets. The research not only assesses effects of canola meal on growth performance and feed utilisation but also investigate effects of canola meal use on fish health status using broad approach including plasma chemistry, histology and gene expression analysis. The present findings provide practical information for barramundi's diet formulation using canola meal as an additional plant ingredient. These also provide implications of potential health effects relating to using canola meal for barramundi. [ABSTRACT FROM AUTHOR]

The morphological and functional development of the digestive system in meagre ( Argyrosomus regius, Asso, 1801) was described by means of histological and enzymatic approaches in order to provide insight into the digestive physiology of the species at early life stages of development and evaluate whether the acquisition of digestive capacities matched the morphological development of digestive organs. For this purpose, A. regius were reared from hatching to the juvenile stage at 18 °C using a standard feeding sequence of enriched rotifers (2–15 days posthatching, dph), Artemia metanauplii (14–51 dph) and microdiet (35–51 dph). Histological and enzymatic analyses indicated that A. regius larvae had at the onset of exogenous feeding (2 dph, 3.2 ± 0.1 mm in standard length, SL) a well differentiated exocrine pancreas, and bile salt-activated lipase and alkaline proteases being the main pancreatic enzymes involved in food digestion. Ontogenic changes in pancreatic enzymes occurred between 3.2 and 3.9 mm in SL, coinciding with the resorption of the oil globule and the complete transition to exogenous feeding, and between 4.2 and 5.8 mm in SL (20–25 dph). Regarding acid digestion, pepsin was not detected in A. regius until 6.0 and 6.8 mm in SL (31 dph), coinciding with notochord flexion and the progressive decrease in the activity of alkaline protease and leucine-alanine peptidase (intestinal cytosolic enzyme) activities, indicating a shift in the mode of digestion. The detection of pepsin activity in A. regius occurred after the appearance of the first gastric glands (4.6 and 5.8 mm SL). Thus, authors need to be cautious when extracting conclusions about the beginning of acid digestion and the onset of weaning of larvae just using histological data, since it has been shown that morphology does not match functionality with regards to the stomach. The comparison of present results with other studies conducted in this species using different rearing conditions ( e.g. mesocosm technique, larval density, water temperature, feeding sequence) indicated that the functional development of the digestive system in A. regius assessed by the activity of alkaline and acid proteases is a well-conserved process that generally occurs within a range of body size regardless of larval age and rearing conditions. Thus, authors recommend using morphometric and/or morphological variables ( e.g. length, developmental stage) together with the age of larvae expressed in days after hatching for comparative purposes among different studies. Statement of relevance The morphological and functional development of the digestive system in meagre ( Argyrosomus regius ) was described by means of histological and enzymatic approaches in order to provide insight into the digestive physiology of the species at early life stages of development. Combining histological and enzymatic analytical procedures we evidenced that stomach's functionality did not match its morphological organization, since pepsin activity was not detected until some days after the appearance of the first gastric glands. This indicates that authors need to be cautious when extracting conclusions about the beginning of acid digestion and the onset of weaning of larvae just using histological data. [ABSTRACT FROM AUTHOR]

Digestive metabolism is considered key to resilience of fish populations as it determines energy and nutrient availability for growth and survival. In cleaner fishes, digestion performance also influences the amount and the rate at which parasites can be removed from co-operating fishes, called hosts. Therefore, understanding the effect of temperature on digestive metabolic scope ( i.e. the energy allocated to digestive processes) is crucial to predicting responses of fish communities to ocean warming. Body size can affect many physiologic processes and is thought to decrease with increasing temperature; therefore, we examined the effect of body mass and warming on digestive metabolic scopes in two sister species of cleaner gobies of the genus Elacatinus that reach different adult sizes. The dwarf-size Elacatinus lobeli increased digestive metabolic rates and scope while the larger Elacatinus oceanops decreased digestive metabolic scope with warming. Intra-specifically, larger E. lobeli also showed a decreased scope when compared to smaller individuals. Results from this study suggest that perhaps smaller fishes may have a digestive and metabolic advantage at higher temperatures and may be more resilient under warming temperatures. [ABSTRACT FROM AUTHOR]

The expression and digestive activity of pike silverside Chirostoma estor endogenous chitinases were analysed in samples from four life stages: whole eggs; larvae; juvenile intestine and hepatopancreas and adult intestine and hepatopancreas. A chitinase cDNA was cloned and partially sequenced ( GenBank accession number: FJ785521). It was highly homologous to non-acidic chitinase sequences from other fish species, suggesting that it is a chitotriosidase. Quantitative PCR showed that this chitinase was expressed throughout the life span of C. estor, with maximum expression in the hepatopancreas of juveniles. Chitotriosidase and chitobiosidase activities were found at all life stages, along with a very high level of N-acetyl glucosaminidase ( NAGase). The chitotriosidase activity could be encoded by the cloned complementary (c) DNA, although additional chitinase genes may be present. The chitotriosidase activity appeared to be transcriptionally regulated only at the juvenile stage. The expression and activity of chitinases tended to increase from the early to juvenile stages, suggesting that these variables are stimulated by chitin-rich live food. Nevertheless, the feeding of juvenile and adult fish with both live food and a balanced commercial diet seemed to provoke significant reductions in pancreatic NAGase secretion and/or synthesis in the gut. Moreover, all chitinase activities were lower in adults, probably reflecting a higher intake and use of the balanced diet. The observation of chitotriosidase and chitobiosidase activities together with a very high NAGase activity suggest the presence of a complete and compensatory chitinolytic chitinase system that enables this stomachless short-gut fish species to use chitin as an energy substrate. These novel findings suggest that dietary inclusions of chitin-rich ingredients or by-products might reduce the farming costs of C. estor without impairing performance. [ABSTRACT FROM AUTHOR]

The prevalence of extreme, short-term temperature spikes in coastal regions during summer months is predicted to increase with ongoing climate change. In tropical systems, these changes are predicted to increase the metabolic demand of coral reef fish larvae while also altering the plankton communities upon which the larvae feed during their pelagic phase. The consequences of these predictions remain speculative in the absence of empirical data on the interactive effects of warm temperatures on the metabolism, postprandial processes and growth responses of coral reef fish larvae. Here, we tested the effect of increased temperature on the metabolism, postprandial performance and fine-scale growth patterns of a coral reef fish (Amphiprion percula) in the latter half of its ~11-d larval phase. First, we measured the length and weight of fed versus fasted larvae (N = 340; mean body mass 4.1±0.05 mg) across fine temporal scales at a typical current summer temperature (28.5°C) and a temperature that is likely be encountered during warm summer periods later this century (31.5°C). Second, we measured routine metabolic rate (Mo2 routine) and the energetics of the postprandial processes (i.e., digestion, absorption and assimilation of a meal; termed specific dynamic action (SDA)) at both temperatures. Larvae fed voraciously when provided with food for a 12-hour period and displayed a temperature-independent increase in mass of 40.1% (28.5°C) and 42.6% (31.5°C), which was largely associated with the mass of prey in the gut. A subsequent 12-h fasting period revealed that the larvae had grown 21.2±4.8% (28.5°C) and 22.8±8.8% (31.5°C) in mass and 10.3±2.0% (28.5°C) and 7.8±2.6% (31.5°C) in length compared with pre-feeding values (no significant temperature effect). Mo2 routine was 55±16% higher at 31.5°C and peak Mo2 during the postprandial period was 28±11% higher at 31.5°C, yet elevated temperature had no significant effect on SDA (0.51±0.06 J at 28.5°C vs. 0.53±0.07 J at 31.5°C), SDA duration (6.0±0.6 h vs. 6.5±0.5 h), or the percent of total meal energy used for SDA (SDA coefficient: 10.1±1.3% vs. 13.0±1.7%). Our findings of higher Mo2 routine but similar SDA coefficient at high temperature provide the first empirical evidence that coral reef fish larvae may have to secure more food to attain similar growth rates during warm summer periods, and perhaps with chronically warmer conditions associated with climate change. [ABSTRACT FROM AUTHOR]

Concentrations of the paralytic shellfish toxins C1+2, C3+4, GTX5, GTX6, dcGTX2+3, dcSTX, dcNEO, GTX2+3, GTX1+4, STX and NEO were determined by LC-FLD in composite samples of digestive glands and gills of Cerastoderma edule cockle. The specimens were sampled in Aveiro lagoon, Portugal, under natural depuration conditions (days 0, 8, 12, 14, 19, 21 and 25) after exposure to a bloom of Gymnodinium catenatum . Individual paralytic shellfish toxins indicated different pathways of elimination and biotransformation in digestive gland and gills. Toxin concentrations in gills were lower than in digestive gland. Most of the quantified toxins in digestive gland decreased during the 25 days of observation according to negative exponential curves, and only GTX5, GTX6 and NEO showed slight irregularities with time. Concentrations of C1+2, C3+4 and dcGTX2+3 in gills decreased progressively, however GTX5, GTX6 and dcSTX showed pronounced increases. Higher concentrations of those toxins in days 8 and 12 in comparison to the initial value (day 0) indicate conversion of other toxins into GTX5, GTX6 and dcSTX during those periods. It appears that inter-conversion of toxins occurs as G . catenatum cells are retained in gills before being transferred to other compartments. [ABSTRACT FROM AUTHOR]

Symptoms indicating digestive malfunction have been observed in cultivated salmon over the last 10–15 years, at least partially as a consequence of use of new feed ingredients. The present knowledge on many aspects of digestive functions in fish is limited, in particular for Atlantic salmon. Strengthening is needed to understand the mechanisms underlying challenges to gastrointestinal health and to find means to prevent their development. The present paper supplies information regarding digestive enzyme optima and stability, chyme pH and enzyme activities along the digestive tract of Atlantic salmon. The data presented includes new data from four studies on Atlantic salmon as well as a review of comparative literature from various other fish studies. The four experiments comprised salmon of different sizes, kept in salt or fresh water, and fed high fish meal diets with different protein to energy ratios (P/E). These studies showed an average pH in the stomach chyme, i.e. across diets and experiments, of 4.8 (SD = 0.7), whereas the observed pH optimum for pepsin was 3.0. In the intestinal chyme, pH increased throughout the intestine from a mean of 8.1 (SD = 0.23) in the proximal intestine to 8.4 in the mid intestine (SD = 0.27) and distal intestine (SD = 0.29). Total proteolytic activity of chyme in the proximal intestine, measured as the sum of the work of all proteolytic enzymes on casein as substrate, showed its maximum at pH 8.8, i.e. clearly above prevailing chyme pH. For the individual proteolytic enzymes, pH optimum varied from 7.10 for elastase 1 to 8.98 for trypsin. Chyme pH did not appear to be influenced by fish size, but was significantly higher in the fish raised in fresh water than in those raised in salt water. In the intestinal chyme pH showed a significant negative regression on diet P/E. For the investigated pancreatic enzymes in the intestinal chyme, the activity in the chyme decreased from proximal to distal regions and faecal activities were very low compared to the activities in the proximal intestine. The activity profile for leucine aminopeptidase (LAP), however, trended to increase from mid to distal intestine. Chyme enzyme activities along the intestinal tract did not differ clearly between the experiments and no clear relationship between P/E on enzyme activities along the intestinal tract was observed. Remarkable in vitro stability was observed for the proteolytic enzyme activities. [ABSTRACT FROM AUTHOR]

The aim of this study was to determine the impact of diets with different plant protein ingredients on proximate composition, tissue metabolites and tissue fatty acid composition, as well as hepatic and intestinal histological modifications in Senegalese sole ( Solea senegalensis). Fish (21.5 ± 2.8 g body weight) were fed six isonitrogenous and isoenergetic diets during 11 weeks. A control diet containing fish meal as the main protein source was compared with five experimental diets replacing 30% fish meal protein by different plant protein sources: soybean meal ( SBM), soybean protein concentrate ( SPC), soybean protein isolate ( SPI), wheat gluten meal ( WGM) or pea protein concentrate ( PPC). The inclusion of different plant protein did not significantly affect growth and proximate composition of fish. The hepatosomatic index was not significantly different to the control group; however, utilization of WGM significantly increased hepatocyte size. Plasma protein values and muscle triglycerides were influenced by the inclusion of SBM and WGM in the diets respectively. Feeding fish on SBM, WGM and PPC diets significantly enhanced n-6 fatty acid levels in muscle, particularly linoleic acid. None of the plant protein ingredient used in the diets decreased arachidonic, eicosapentaenoic as well as docosahexaenoic acid values in liver and muscle. Overall, histological studies did not reveal the existence of any intestinal alterations induced by the inclusion of different plant proteins. Despite moderate changes produced by SBM, SPC and WGM, inclusion of dietary plant protein ingredients has no major impact on growth, tissue and blood biochemistry, fatty acid profile and gut integrity of Senegalese sole juveniles. [ABSTRACT FROM AUTHOR]

This study investigates the effects of gradual or abrupt changes in rearing salinity on food transit time and digestive enzymes activity of Caspian kutum (Rutilus kutum) larvae. The larvae (532 ± 0.05 mg) were supplied and randomly allocated into 12 tanks at a density of 45 fish per tank. Experimental treatments were fresh water (salinity 0) [FW] as control, exposure to salinity 5 [T1], and gradual transfer to salinity 10 in two steps of first to 5 h, then and after 12 h to a salinity of 10 [T2], and abrupt change (direct transfer to a salinity of 10 [T3]). Results showed at 8 h after start of feeding that the larvae intestine was filled with food pellets except in treatment T1. Enzyme activity responded to salinity change as follows: the highest trypsin, amylase, and chymotrypsin activities were observed in T1; however, these were not significantly different to treatment T3 (P > 0.05). Trypsin activity peaks in the FW and T2 groups occurred 8 h after feeding, and in T3 and T1 groups 5 h after feeding. Peak chymotrypsin and alkaline phosphatase activity was observed 5 and 8 h after feeding in all experiments, respectively. The highest a amylase activity in FW and T2 groups occurred 5 h after feeding, while in T3 and T1 these peaks were observed 8 h after feeding. These results indicate that salinity had some noticeable effects on the activities of digestive enzymes after feeding. [ABSTRACT FROM AUTHOR]

The structure and histochemical properties of the intestine in bronze corydoras ( Corydoras aeneus), a stomach-containing teleost, are described, with emphasis on goblet cells and mucin types. The proximal intestine displayed a normal structure for teleosts, whereas the distal intestine was wide, translucent, thin-walled, richly vascularized and constantly filled with air, suggesting an important respiratory role. Goblet cells were common throughout the entire intestine and displayed a variable, but mainly faint metachromatic colour after toluidine blue. They were moderately coloured by alcian blue at both pH 2.5 and 0.2 and displayed no colour after periodic acid followed by Schiff's solution (PAS), but a distinct purple-brown colour after high iron diamine followed by alcian blue (p H 2.5). Together, these results suggest that the mucin in the intestine goblet cells consists mainly of sulphated proteoglycans. Further, the results from the present lectin and neuraminidase tests suggest that these mucins contain much N-acetylglucoseamines and some N-acetylgalactosamines and sialic acid, but seem to lack glucose and mannose. They also contain some galactose- N-acetylgalactosamines sequences, normally hidden by sialic acid. The distinct brush border and mucus layer on the epithelial cells in the respiratory intestine may indicate some digestive roles, such as absorption of water, ions and simple carbohydrates. As sulphated proteoglycans are tough and attract much water, this mucus may play important roles in the protection against mechanical and chemical damages and in the defence against micro-organisms throughout the entire intestine, but in the respiratory intestine it may impede significantly the oxygen uptake. However, as this part of the intestine usually contains no digesta, but is completely filled with air, frequently renewed by dry air from the atmosphere, and the main function of the mucus may be to protect the respiratory epithelium against a destroying and dangerous desiccation. [ABSTRACT FROM AUTHOR]

The present study was conducted to elucidate the effects of copper exposure on the immune system and lipid metabolism of the Japanese scallop, Mizuhopecten yessoensis . Transcriptional levels of differentially expressed genes (DEGs)in M. yessoensis digestive gland tissue were analyzed using the deep-sequencing platform Illumina HiSeq™ 2000. In total, 841 and 877 genes were identified as significantly up- or down-regulated, respectively. In addition, significant enrichment analysis identified 3 gene ontology terms and 15 pathways involved in the response to copper exposure. Analysis of transcripts related to the immune response revealed a complex pattern of innate recognition receptors, including toll-like receptors, NOD-like receptors and downstream pathway effectors, including those involved in apoptosis. Furthermore, genomic analysis revealed that genes involved in extracellular matrix (ECM)-receptor interactions were enriched in Cu-exposed scallop glands. These results will provide a resource for subsequent gene expression studies regarding heavy metal exposure and the identification of copper-sensitive biomarkers for the aquaculture of M. yessoensis . [ABSTRACT FROM AUTHOR]

Trypsin is an important serine protease that is considered to be involved in digestion of protein in teleost fish. Nevertheless, studies on trypsin/trypsinogen in fish embryos are very limited. In this study, the trypsinogen of turbot ( Scophthalmus maximus ) ( tTG ) was identified and the expression patterns and activity of trypsinogen/trypsin were investigated. The results showed that the tTG mRNA was evenly distributed in the oocytes and was also expressed along the yolk periphery in early embryos. At later embryo stages and 1 days after hatching (dph), the tTG mRNA concentrated at the alimentary tract and head. Quantitative expression analysis showed that the tTG transcripts decreased after fertilization until the gastrula stage, then increased with the embryo and larvae development. This result was also confirmed by the specific activity analysis of trypsin and in-situ-hybridization (ISH). All of the results indicated that tTG in early embryo stages was maternally derived and expressed by itself after gastrula stages. Additionally, location of tTG mRNA in embryos and larvae was investigated; we considered that trypsin may have multiple functions during the embryo development process. Based on our results regarding trypsinogen in embryos and early development, we concluded that the trypsin/trypsinogen in turbot embryos was inherited from a maternal source and we suggested that trypsin in early development has multiple functions in the process of development. [ABSTRACT FROM AUTHOR]

The digestive tract of Trachelyopterus striatulus (Steindachner, 1877) was studied using morphological, histological, and histochemical techniques. The barbels, lips and tongue had stratified squamous epithelium with mucous, claviform cells, and taste buds. Trachelyopterus striatulus had a dental plaque with villiform teeth. The bucopharyngeal cavity was formed by the gill apparatus and pharyngeal teeth with vilifform denticles. The oesophagus presented stratified squamous epithelium with mucous cells and taste buds. The stomach included cardiac, fundic, and pyloric regions and a simple prismatic epithelium with prismatic cells that reacted positively to periodic acid-Schiff (PAS) and amylase+PAS. Only the cardiac and pyloric regions responded positively to alcian blue pH 2.5 (Ab pH 2.5) and alcian blue pH 0.5 (Ab pH 0.5). The cardiac and fundic regions exhibited tubular gastric glands. The intestine was 118.90 ± 22.49 mm long with an intestinal coefficient (CO) of 0.83 ± 0.13. The epithelium was simple prismatic with a brush border and goblet cells, and a greater number of goblet cells were found in the caudal region of the intestine. The mucous cells and goblet cells reacted positively to PAS, amylase+PAS, Ab pH 2.5, and Ab pH 0.5. We analysed if the function of the mucosubstances and morphological characteristics of the digestive tract of T. striatulus are compatible with omnivorous feeding habit. [ABSTRACT FROM AUTHOR]

A 90-day feeding trial was conducted to determine the effects of dietary organic selenium ( OS) supplementation on immune responses, total selenium accumulation in various tissues and the digestive system health of marron Cherax cainii. A group of marron (initial weight 3.65 ± 0.05 g) were fed 0.2 g kg−1 of Sel-Plex® as a source of OS and were compared to a control group of marron that were not fed OS. After 90 days of feeding OS, total haemocyte counts ( THC) and the percentages of granular and semigranular cells in marron were significantly higher than in marron fed a control diet. However, dietary OS did not affect the proportion of hyaline cells. The marron fed OS had significantly lower bacteraemia and longer neutral red retention times than the control group. Total selenium levels in the haemolymph, hepatopancreas and muscle tissues of the OS-fed marron group were significantly higher ( P < 0.05) than in control diet group. The highest total soluble selenium levels were found in the muscle tissues of OS-fed marron on the final day of the feeding trial. Amylase and protease activity, soluble protein, and numbers of microvilli were significantly higher ( P < 0.05) in marron fed OS. These findings demonstrate the benefits of including 0.2 g kg−1 OS in marron diet to enhance immune parameters, increase total selenium accumulation in muscle and improve marron digestive system health. [ABSTRACT FROM AUTHOR]

This study was undertaken to address a lack of understanding of the early developmental stages of the tropical elasmobranch whitespotted bamboo shark,Chiloscyllium plagiosum, to assist with advancement of its aquaculture. It is the first report of the histological and ultrastructural characteristics of its developing digestive tract. The study was carried out from hatching to 60 days after hatching (DAH). Embryos for this study were maintained in the laboratory. Histological investigations yielded the following data. At hatching, the gut of an embryo consisted of an undifferentiated straight tube that lay over the yolk sac. The digestive tract was differentiated into buccopharynx, esophagus, stomach, and intestine by 8 DAH. During the preflexion phase (within 16 DAH), the mucous cells of the esophagus were differentiated. Gastric glands were present at 60 DAH, indicating the transition from the embryonic to the juvenile stage and the acquisition of an adult mode of digestion. The lumen of the intestine appeared on 6 DAH; the intestinal valve appeared and divided the intestine into anterior and posterior sections on 20 DAH. The liver and the pancreas were formed on 8 DAH. The yolk sac was partially depleted by 80 DAH. In terms of ultrastructure, on 30 DAH, the anterior section of the esophagus was lined by a stratified epithelium, whereas the posterior section was lined by simple tall columnar epithelial cells. Goblet cells and oxynticopeptic cells were found in the stomach. The pancreatic acinar cells were strongly elongated and of two types: one with a pale and the other with a more opaque cytoplasm. [ABSTRACT FROM AUTHOR]

In this study, we investigated the effects of enzyme-producing probiotic bacteria isolated from the gastrointestinal tract of rainbow trout on the growth performance, feed conversion ratio, and digestive enzyme activity, of fry (Oncorhynchus mykiss). Three isolates (G8/2013, T7/2013 and U5/2013) of candidate bacteria elicited the highest protease, lipase, and amylase activities, respectively. Isolates were identified as Aeromonas sp., Bacillus sp. and Citrobacter braakii by morphological, physiological, biochemical characterizations as well as 16S rRNA gene sequence analysis. The fry basal diet was supplemented with probiotics at varying concentrations; G8 group, Aeromonas sp. 1.72 × 108 CFU/g; U5 group, Bacillus sp. 3.01 × 108 CFU/g; T7 group, C. braakii 2.96 × 108 CFU/g and a mixed group (same bacterial concentrations), and control group (no bacteria). The rainbow trout fry were fed ad libitum in triplicate treatments with supplemented and non-supplemented probiotic diets for 70-days. The total bacterial count in the intestine was significantly higher in the mixed group (30th and 50th days) and U5 group (50th and 70th days) compared with the control group. However, there was no significant difference in weight gain, specific growth rate (SGR), feed conversion ratio (FCR), nutrient digestibility, or digestive enzyme activity among the groups. [ABSTRACT FROM AUTHOR]

A commercial diet (C) (48% protein and 20% lipids) and a high-digestible carbohydrate diet (CH) (37% protein, 12.5% lipids and 40% high-digestible carbohydrates) were used to feed sea bream juveniles for an 8-week period. In the commercial diet, more than 60% of ingredients were of plant origin from various sources, whereas the only component of plant origin in the CH diet was wheat. To determine the best time to administer carbohydrates and the possible protein-sparing effect, three different dietary regimes were established: C, CH-M and CH-A, and the corresponding diet was fed to sea bream in the morning (1.6% of bw) and in the afternoon (1% of bw), calculating quantities according to the amount of feed that fish ate during the acclimatization period. After the growth trial, specific growth rate (SGR), relative intestinal length, intestinal pH content, gastric and pancreatic digestive enzyme activities and nutrient absorption capacities were studied 5 h post-feeding after each meal (morning and afternoon). The acid protease activity measured was anticipatory and was higher when the next meal would have more protein. No differences in relative intestinal length or feed buffering capacity were found. The smaller ration given to sea bream in the afternoon led to a lower pancreatic release of alkaline protease and α-amylase and an up-regulation of D-Glc and L-Ala absorption capacity. A higher transit rate was measured when sea bream were fed the CH diet. When high-digestible carbohydrates were administered in the morning and the commercial diet in the afternoon, we observed a better assimilation of both diets due to compensatory mechanisms such as an increase in L-Lys, D-Glc and L-Ala absorption capacity after the morning feed, and a higher pancreatic release of alkaline protease and amylase after the afternoon feed. In contrast, when high-digestible carbohydrates were given in the afternoon, only a significant up-regulation of the capacity to absorb L-Lys was detected. Thus, the inclusion of high-digestible carbohydrates in the diet improved digestion and absorption processes when administered in the morning, leading to a protein-sparing effect that yielded growth comparable to that of fish fed an exclusively commercial diet. [ABSTRACT FROM AUTHOR]

In this work, histological and histochemical features of the larval digestive system of Argentine anchovy Engraulis anchoita were described. Structural changes during ontogenetic development were also characterized, and comparisons between the beginning and the end of larval development were made. Histological sections of larvae were subjected to histochemical and routine histological techniques to localize and differentiate glycoproteins ( GPs). Both at an early and a late larval stage, the oesophageal goblet cells reacted more intensely than those of the rest of the digestive tract, and only the oesophagus exhibited metachromasia with toluidine blue techniques at different pHs, thus revealing diverse GPs at different concentrations. The GPs histochemical composition in the intestine varied with the developmental stage and the intestinal zone. The absence of goblet cells characterized the foregut; however, they started differentiation at an advanced stage in the midgut. These cells could be detected in the hindgut both at the beginning and at the end of development. The attached glands showed a varied glycoprotein composition. The digestive tract of E. anchoita presented a high level of complexity, related to the multiple functions of mucus in the digestive tract, such as lubrication, protection, antimicrobial function and ionic and osmotic regulation. [ABSTRACT FROM AUTHOR]

The effect of burbot weight ( BW) and length ( TL) on the efficiency of weaning under controlled conditions was investigated. Growth, survival rate, cannibalism and histological analysis of digestive tract were examined. The experiment lasted 49 days (40-89 days post hatch [ DPH]). The fish ( BW 0.12 g; TL 14.1 mm) were divided into four groups: a control group ( C) exclusively fed Artemia sp. nauplii and groups F40, F47 and F54 that were weaned to dry feed on 40, 47 and 54 DPH, respectively. The highest survival rate was observed in groups C (78%) and F54 (58%). In group F54, similar TL (53.8 mm) and higher BW (1.36 g) at the end of experiment, in comparison with group C (43.9 mm, 0.84 g for TL and BW, respectively), were obtained. At the end of the experiment, the length of enterocytes in groups C and F54 was at a similar level (over 23 μm), whereas in group F47 was significantly lower (approximately 20 μm; P < 0.05). This study for the first time presents successful weaning of the burbot. The data indicate that live food should be offered until the fish reach 25 mm and 0.20 g. [ABSTRACT FROM AUTHOR]

This is the first study investigating the plant-herbivore interaction between Sarpa salpa, which has overgrazed seagrass transplants in Portugal, and the seagrasses Cymodocea nodosa, Zostera marina and Zostera noltii, which have been considered for restoration. When offered the choice between the three seagrasses in outdoor tanks, adult S. salpa clearly preferred Z. noltii. Testing the seagrasses separately, mean ± s.d. feeding rates ranged from 21 ± 11 g seagrass fresh mass kg−1 fish mass day−1 for Z. marina to 32 ± 9 g seagrass fresh mass kg−1 fish mass day−1 for C. nodosa and 40 ± 11 g seagrass fresh mass kg−1 fish mass day−1 for Z. noltii (temperature = 16° C). Food-processing rate in S. salpa did not differ between seagrasses, and there was no evidence of a regulation of processing rate according to food intake. Seagrasses differed substantially in nitrogen content and C:N, with C. nodosa containing the highest nitrogen content and lowest C:N (2·5 ± 0·1% and 14·0 ± 1·0), followed by Z. noltii (2·1 ± 0·1% and 17·0 ± 1·0) and Z. marina (1·4 ± 0·1% and 26·0 ± 2·0). Food-processing rate in S. salpa and the nutritional value of the seagrasses were not correlated with the observed feeding preference and rate. The study suggests that C. nodosa and Z. marina are less at risk of overgrazing by S. salpa and might thus be preferable to Z. noltii for seagrass restoration in areas with noticeable abundances of this fish. [ABSTRACT FROM AUTHOR]

Modern feeds for industrial salmon farming are presently formulated to fulfill specific digestible protein (DP, grams per kg feed, hereafter called units) and digestible energy (DE, Mega Joule (MJ) per kg feed) specifications. In the present study, 299 feed samples and accompanying fecal samples were obtained from 43 different farming sites for Atlantic salmon along the coast of Norway between October 2010 and February 2013. Apparent digestibility coefficients for protein and fat were calculated from protein and fat in feed and fecal matter, using analyzed crude fiber contents as a digestibility marker. Overall mean apparent protein and fat digestibilities were 87.1% (±4.9; overall mean±S.D.) and 92.5% (±3.7), respectively. Linear mixed models with rearing site as random effect were applied to evaluate factors that influenced the apparent digestibility of protein and fat, feeding intensity and energy intake. Apparent digestibility of fat tended to increase over the study period, and both feed type and feed manufacturing company, and water temperature influenced the digestibility significantly. Digestible energy content (DE, MJ/kg) was significantly negatively associated with feeding intensity, but not with total digestible energy intake. Variation at site level was significant in all analyses, and was moderate for the digestibility analyses and high regarding feeding intensity and energy intake. Digestibility assessments were used to assess accuracy, precision and agreement between optimized and realized digestible protein (DP, %) and DE level of feeds, using the mean difference between the two sets of observations and a fixed contribution of dietary energy from starch. Overall mean difference between realized and optimized DP and DE was −0.28units and −0.56MJ/kg, respectively, and there were significant differences between feed manufacturing companies. The results indicated that field digestibility assessments can be used to validate nutritional and economical value of feeds in commercial salmon farming. [ABSTRACT FROM AUTHOR]

Abstract: The effects of dietary probiotics on growth performance, health status of the digestive system and on the blood parameters of juvenile rainbow trout (Oncorhynchus mykiss, Walbaum) were investigated. Compared to the controls, diets supplemented with Kocuria SM1 and Rhodococcus SM2 for 14days caused no significant (P>0.05) change on the growth of fish, although SM2 led to an increased specific growth rate. SM1 or SM2 started to colonize in the gut after 7days of administration reaching a maximum, i.e. ~90–100% of total bacterial population, at 14days. Yet, the probiotics disappeared from the gut once administration ceased. Structural differences were not noted in the pyloric caeca and foregut, but the overall condition of the midgut and hindgut improved in fish receiving probiotics. The profile of digestive enzymes, as determined by the use of API ZYM, and stress indicators, i.e. serum hemoglobin, urea, creatinine and glucose, was not affected by any of the experimental diets (P>0.05). [Copyright &y& Elsevier]

The latitudinal distribution of persistent organic pollutants (POPs: legacy organochlorines [OCs], polybrominated diphenyl ethers [PBDEs,] and hexabromocyclododecane [HBCD]) was examined in livers of two species of marine fish, the pelagic saithe (Pollachius virens,n = 40) and the demersal cod (Gadus morhua,n = 40), along a south-north gradient (59°-70°N) on the Norwegian Coast. Cod had in general two to three times higher concentrations of POPs than saithe, probably because of higher exposure in the benthic food chain. The concentrations of heavy halogenated compounds were higher in the southernmost region than further north. Moreover, the POP pattern showed a gradual shift in the compositions from south to north, especially for OCs in cod: i.e. the relative importance of low-chlorinated polychlorinated biphenyl (PCB) congeners and some OC-pesticides (e.g., hexachlorobenzen [HCB]) in the contaminant burdens increased with latitude. The latitudinal fractionation signal was weaker in saithe, possibly due to its pelagic and nomadic behavior. Hence, this study shows not only a strong latitudinal fractionation in the compositional patterns of POPs in marine fish but also the effects of habitat use and fish behavior. [ABSTRACT FROM AUTHOR]

Abstract: Chymotrypsin from the hepatopancreas of cuttlefish (Sepia officinalis) was purified to homogeneity, with a 120-fold increase in specific activity and 23% recovery. The molecular weight of the purified chymotrypsin was estimated to be 28kDa by sodium dodecyl sulphate–polyacrylamide gel electrophoresis. The optimum pH and temperature for the chymotrypsin activity were pH 8.5 and 55°C, respectively, using succinyl-l-ala-ala-pro-l-phenylalanine-p-nitroanilide (SAAPFpNA) as a substrate. The enzyme was extremely stable in the pH range of 7.0–10.0 and highly stable up to 50°C after 1h incubation. This proteinase was strongly inhibited by chymostatin, soybean trypsin inhibitor, diisopropylfluorophosphate and phenylmethylsulfonyl fluoride, but was not inhibited by tosyl-l-phenylalanine chloromethyl ketone, N-carbobenzoxy-phenylalanine chloromethyl ketone or N α-tosyl-l-lysine chloromethyl ketone. The enzyme hydrolysed long chymotrypsin peptide substrates SAAPFpNA, SAAPLpNA and ZAALpNA and did not hydrolyse short chymotrypsin substrates. Kinetic parameters of the enzymatic reaction demonstrated that the best substrate was SAAPFpNA, with k cat 18s−1 and K m 22μM. However, the enzyme had a lower K m for SAAPLpNA, 54μM. The N-terminal amino acid sequence of the first 20 amino acids of the purified chymotrypsin was IVGGQEATIGEYPWQAALQV. [Copyright &y& Elsevier]

Abstract: Proper nutrition at first feeding in marine fish larvae is an important factor for successful larval rearing. However, live prey used to feed marine finfish larvae lack essential fatty acids requiring commercial hatcheries to enrich live prey to provide a balanced diet. Currently, the many commercially available enrichments for live prey present considerable differences in their formulas, and compositions of essential nutrients. Since nutritional requirements are species-specific it is necessary to define those enrichments most appropriate for each species in terms of larval development and quality. This study aimed to evaluate the effect of six enrichments on Senegalese sole larval performance including growth, digestive system development and skeletogenesis, to determine whether any of these products are more suitable for Senegalese sole larval rearing. From 8 to 37days post hatching (dph), larvae were fed Artemia nauplii previously enriched with six different formulae: Easy Selco© (INVE, ES), Easy Selco© (INVE) half diluted with olive oil (ES/2), Multigain© (BioMar, MG), Red Pepper© (Bernaqua, RP), Aquagrow Gold© (ABN, AGG) and Aquagrow DHA© (ABN, AGD), with the last two being diluted by a third with olive oil. The dietary treatments significantly affected larval growth and performance; larvae fed Artemia enriched with AGG presented significantly higher final dry weight (2.0±0.4mg), growth and intestinal maturation than larvae fed ES-enriched Artemia (final dry weight: 1.1±0.3mg). Larvae fed the AGG treatment also were those most advanced in metamorphosis and ossification processes. Larvae fed ES/2, RP, AGD and MG treatments presented intermediary values for all these parameters. No significant effect of the dietary treatments was detected in final survival and incidence of skeletal deformities. We conclude that, among the enrichments tested, AGG is the most appropriate for larvae of Senegalese sole. [Copyright &y& Elsevier]

Abstract: The appearance of stomach pepsin activity in teleosts is one of the last ontogenetic events during the larval stages, and typically marks the completion of the shift from intracellular to extracellular mode of digestion. The present trial describes the morphological and functional ontogeny of cobia''s stomach. Larvae were reared in a recirculated system and fed enriched rotifers in green water, followed by enriched Artemia, prior to being weaned on an artificial diet 25days-post-hatch (dph). The stomach commenced differentiation at 6dph, and displayed numerous gastric glands by 16dph (11.1mm TL). However, pepsin-like activity was not detectable prior to 22dph (20.8mm TL, P<0.05), denoting a lack of functionality until this time point. Maximum pepsin-like activity reached 29.09±1.47UHb larvae−1. Together with the observations of others, the results presented herein shed light on the ontogeny of proteolitic digestion in cobia. [Copyright &y& Elsevier]

In bay snook ( Petenia splendida) larvae the histological development of the digestive system and swim bladder, and their relative timing of differentiation were studied from hatching to 45 days post-hatch (dph) at 29°C. Newly hatched larvae showed a simple digestive tract, which appeared as a straight undifferentiated tube lined by a single layer of columnar epithelial cells (future enterocytes). The anatomical and histological differentiation of the digestive tract and accessory glands was a very intense, asynchronous process, proceeding from the distal to the anterior part. The intestine was the first region to differentiate (9 days post-hatch - dph, 6.5 mm SL), and the oesophagus the last (21 dph, 8.4 mm SL). At the onset of feeding, the digestive system was organized into different functional and histologically differentiated sections, such as the buccopharynx, oesophagus, glandular stomach, and anterior and posterior intestine. This organization resembled that of the juveniles, with the exception of pharyngeal teeth and buccopharyngeal as well as oesophageal goblet cells, which proliferated later during the mixed feeding period. Histological observations revealed that bay snook larvae retained endogenous yolk reserves until 24 dph (8.9 ± 0.4 mm SL), which might be helpful for weaning this species onto a compound diet. The important lipidic accumulation observed in the intestinal mucosa, liver, and pancreas in fish fed a compound trout diet indicated that although fish were able to digest and absorb lipids, the diet formulation did not fit the nutritional requirements of early juveniles of this species. The ontogeny of the digestive system followed the same general pattern as in most cichlid species described to date. However, we detected species-specific differences in the timing of differentiation that were related to their reproductive guild. According to the histological results, some recommendations regarding the intensive culture of this species are also provided. [ABSTRACT FROM AUTHOR]

The effects of prolonged feed deprivation (40 days at 18° C) and re-feeding (30 days) on body mass, growth and the activity of selected pancreatic and intestinal enzymes were evaluated in migrating European glass eels Anguilla anguilla by comparison with a control group fed to satiation with hake Merluccius merluccius roe for the duration of the experiment. Feed deprivation resulted in mass loss and a reduction in digestive function, as revealed by a decrease in the total and specific activities of pancreatic (trypsin and α-amylase) and intestinal brush border (alkaline phosphatase and leucine aminopeptidase) enzymes. The total activity of intestinal brush border enzymes diminished after 5 days of feed deprivation, whereas that of pancreatic enzymes did not decrease until 10 days, indicating that the intestine is more sensitive to feed deprivation than the pancreas. Re-feeding A. anguilla that were starved for 40 days resulted in compensatory growth, with specific growth rates that were 2·6 times higher than the control group. This compensatory growth was associated with the recovery of trypsin and intestinal brush border enzyme activities, which were restored to control levels within 5 days of re-feeding. The ability to maintain pancreatic enzyme activity during 40 days of feed deprivation, and rapidly recover capacity for protein digestion upon re-feeding, would enable A. anguilla at this glass eel stage to withstand periods without food but rapidly provide amino acids for protein synthesis and growth when suitable food was available. [ABSTRACT FROM AUTHOR]

In this work, several features during the wedge sole larval development have been described. The newly hatched larva presented an acidophilic yolk with some oil drops. The digestive tract began to differentiate at 1 DAH, with a loop being discernible. The pancreas and liver were completely formed at 2 DAH, the former showing its typical basophilic acinar structure and acidophilic zymogen granules. The first supranuclear vesicles in enterocytes were seen at 3 DAH. At 4 DAH, yolk reserves were completely exhausted, the number of oesophagus and intestine mucous cells increased, and the heart was differentiated into four chambers: the venous sinus, atrium, ventricle, and arterious bulb. The development was fast and almost all organs were differentiated at 2 DAH. It is important to emphasize that gastric glands were not detected, a factor that should be considered when deciding diet formulation and feeding strategies for the rearing of this species. [ABSTRACT FROM AUTHOR]

Abstract: The major neutral lipase excreted by the pancreas in fish, is bile activated lipase (BAL). Here we present evidence that cod have a functional BAL and a non-functional pancreatic lipase related protein (PLRP). The Atlantic cod genome does not seem to contain colipase which is essential for pancreatic lipase activity. During the larval stages, the gene expression of BAL was low until the point when pyloric caeca started to differentiate and develop (~20mm standard length (SL)). Then the expression increased until ~50mm SL. The PLRP gene was expressed but showed very little regulation. The activity of neutral lipase did not increase in parallel to gene expression. The mismatch between activity and gene expression measurements may be partly explained by the unspecific analytical method, when analysing lipase activity in larva whole body. There is neutral lipase activity in numerous tissues in the fish larvae and the lipase activity in the gut, relatively to activity in the whole body, decreased with age. Furthermore, neutral lipase activity in rotifers was ten times higher than in whole cod larvae with full guts. Activity originating from the live prey may therefore explain the high whole body lipase activity from 3 to 20dph. The results also indicate that “adult type” digestion of neutral lipid develops late in the larval period (from 20mm SL), while other mechanisms of lipid uptake are active at the early larval stage. [ABSTRACT FROM AUTHOR]

Abstract: To investigate the effect of dissolved oxygen level ([O2]) on maintenance metabolism, feeding metabolism, aerobic swimming performance and their metabolic interaction in juvenile southern catfish (Silurus meridionalis Chen), we measured the following: (1) the resting oxygen consumption rate (MO2rest) over a range of water [O2] and from this we calculated the critical oxygen tension (P crit) of fasting fish; (2) the postprandial MO2 response (10% body mass meal size) at water [O2] of 1, 2, 4 and 8mgO2 L−1; and (3) the swimming performance of fasting and digesting fish at water [O2] of 1, 2, 4 and 8mgO2 L−1 at 25°C. The MO2rest remained constant over a broad range of water [O2] but then dropped markedly upon reaching the P crit (16.4% saturation). Hypoxic groups presented lower peak postprandial MO2 (MO2peak) (1mgO2 L−1 group), larger energy expenditure and longer digestive process (both 1 and 2mgO2 L−1) than those of normoxic groups. Both critical swimming speed (U crit) and the active metabolic rate (MO2active) of fasting fish remained unchanged over a decrease in water [O2] from 8 to 4mgO2 L−1 and then decreased significantly with further decreases in water [O2]. These parameters in fed fish showed a pronounced decrease as water [O2] decreased from 8 to 1mgO2 L−1. Feeding caused a significantly lower U crit in the 2mgO2 L−1 water [O2] group, a significantly higher MO2active in both the 2 and 8mgL−1 water [O2] groups and a significantly higher metabolic scope (MO2active − MO2rest) in both the 2 and 4mgO2 L−1 water [O2] groups compared to fasting fish. The MO2 increased greatly with swimming speed in the higher water [O2] groups, whereas it leveled off as swimming speeds approached the U crit in the lower water [O2] groups. Within all water [O2] groups, feeding caused a higher MO2 compared to fasting fish when fish swam at the same speeds, except in the 1mgO2 L−1 group. This finding showed that the critical water [O2] for maintenance of metabolism, digestion, swimming and postprandial swimming increased sequentially due to the increasing O2 demand, which challenges respiratory capacity. With the administration of the medium-sized meal that was used in this study, the metabolic mode of juvenile southern catfish changed from an additive mode during normoxia to a digestion-priority mode under moderate hypoxia and to locomotion-priority mode under hypoxia. [Copyright &y& Elsevier]

Abstract: Ration levels were used to determine the digestible protein (DP) and amino acids as well as digestible (DE) and metabolizable energy (ME) requirements of Atlantic salmon post-smolts for maintenance and the efficiency of utilization for gain above maintenance. The salmon (96g) were held in seawater at a temperature of 10°C and were fasted for 28days or fed incremental ration levels of a fishmeal-based diet for 51days. The composition of gain was determined by the comparative slaughter technique. The relationships between intake of digestible or metabolizable nutrients or energy and gain of those nutrients or energy were linear (R 2 ≥0.98) between zero intake and maximum voluntary intake. The DE and ME requirements for maintenance were 31.5 and 30.6kJkg−0.8 d−1, respectively, and the efficiency of utilization of DE and ME for gain were 0.80±0.01 and 0.82±0.01, respectively. The efficiency of utilization for gain of DP was 0.64±0.01 and that of the sum of digestible amino acids 0.57±0.02. The efficiency of utilization of the individual amino acids for gain varied from 0.45 for tryptophan to 0.65 for histidine. The maintenance requirement for DP was 117.5mgkg−0.7 d−1 and for digestible sum of amino acids 204.0mgkg−0.7 d−1. The maintenance requirements for the individual amino acids varied from 4.3mgkg−0.7 d−1 for methionine and tryptophan to 20.9mgkg−0.7 d−1 for leucine. The results suggest more emphasis should be directed towards determining the maintenance requirements and the efficiency of utilization for gain above maintenance for the individual amino acids and the sum of amino acids rather than for crude protein. [Copyright &y& Elsevier]

Abstract: A 10-week feeding trial in seawater floating cages (1.5×1.5×2.0m) was conducted to investigate the effects of dietary canola meal (CM) levels on growth, survival, digestion and selected immune parameters of Japanese seabass (initial average weight 8.3±0.15g). Six isonitrogenous (crude protein 43%) and isoenergetic (20kJg− 1) practical diets were formulated by replacing 0 (the control), 10, 20, 30, 40, and 50% of protein from fish meal with CM. Each diet was randomly fed to triplicate groups of fish, and each cage was stocked with 20 fish. Fish were fed twice daily (06:30 and 16:30) to apparent satiation. The results showed that with increasing dietary CM levels, the survival, specific growth rate (SGR) and feed efficiency (FE) decreased. The survival was significantly lower compared to the control group (P <0.05) with a 40% substitution level. Fish fed the diet with 20% or more protein from CM had significantly lower SGR than the control group (P <0.05), but there was no significant difference in FE at this level (P >0.05). The activities of digestive enzymes and apparent digestibility coefficients of dry matter, protein, lipid and phosphorus significantly decreased with increasing dietary CM level. A similar trend was observed in the immune response where lysozyme activity significantly decreased compared with the control group when the substitution level was more than 20%. Results of the present study indicated that protein from CM could substitute less than 20% for fish meal protein without influencing the growth of Japanese seabass. The higher substitution levels of CM induced negative influences on growth, survival and serum lysozyme of Japanese seabass. [Copyright &y& Elsevier]

Abstract: An enzyme was purified from the pyloric caecum of tambaqui (Colossoma macropomum) through heat treatment, ammonium sulfate fractionation, Sephadex® G-75 and p-aminobenzamidine–agarose affinity chromatography. The enzyme had a molecular mass of 23.9kDa, NH2-terminal amino acid sequence of IVGGYECKAHSQPHVSLNI and substrate specificity for arginine at P1, efficiently hydrolizing substrates with leucine and lysine at P2 and serine and arginine at P1′. Using the substrate z-FR-MCA, the enzyme exhibited greatest activity at pH 9.0 and 50°C, whereas, with BAPNA activity was higher in a pH range of 7.5–11.5 and at 70°C. Moreover, the enzyme maintained ca. 60% of its activity after incubated for 3h at 60°C. The enzymatic activity significantly decreased in the presence of TLCK, benzamidine (trypsin inhibitors) and PMSF (serine protease inhibitor). This source of trypsin may be an attractive alternative for the detergent and food industry. [Copyright &y& Elsevier]

Abstract: Krill growth and physiological condition were measured during the Baseline Research on Oceanography, Krill and the Environment - West (BROKE-West) large-scale survey of the Western Indian Ocean sector (30-80 °E) of the Southern Ocean. Krill growth and condition were related to sea-surface temperature (SST) and Chlorophyll a levels. Estimates of Antarctic krill growth were obtained using the Instantaneous Growth Rate (IGR) technique, which reflects in situ growth of krill during the intermoult period prior to capture. A total of 10,362 krill were incubated in the IGR experiments conducted on board the research vessel. The average growth rate ranged between 1.8–9.6% of body length throughout the study area (n=1557). Juvenile and sub-adult krill showed higher growth rates compared to adults. Growth decreased with size for krill of all maturity classes, with the largest krill (total length >50mm) expressing negative growth. On average, mean growth was higher (6.5%) in the western section (30-55° E) of the study area compared to the east (56-80° E) (4.2%). In the western section significantly higher growth rates were recorded for krill sampled close to the ice-edge (8.4%) compared to the open ocean (4.2%), which may be due to the presence of phytoplankton blooms associated with the retreating sea ice. However, in the eastern sector higher growth rates occurred in krill from the open ocean compared to the krill caught in areas closer to the coast (5.4% and 2.6% respectively). Krill condition was measured in terms of digestive gland size as a percentage of carapace length. The size of krill digestive gland relative to carapace length ranged from 0.46 to 0.66 (n=2403). A significant positive correlation was found between growth rate and size of the digestive gland (r2=0.7), indicating that growth is related to feeding events. Both digestive gland size and krill growth significantly increased with increasing Chlorophyll a levels and declining SST. [Copyright &y& Elsevier]

Abstract: Fatty acid (FA) composition of nine organs from two closely related Antarctic fish species, Notothenia coriiceps and Notothenia rossii, was determined through gas chromatography with flame ionization detection. A data set for each species was obtained using major FA profiles from specimens caught in the sea waters of Admiralty Bay during the summer season. The FA profiles for both species are overall similar, but organ peculiarities have been found, which could reflect metabolic specificities and feeding habits between species. With the exception of liver, the most abundant FA in organs was the n−3 polyunsaturated FA. The total n−6 polyunsaturated FAs were minor components in all evaluated organs. Palmitic acid was identified as the major saturated FA, whereas oleic acid was the most represented of the monounsaturated FA in almost all assessed organs of both species. The n−3/n−6 ratios of all organs were higher than 3.5. Differences in individual FA and FA metabolic profiles of some organs observed between N. coriiceps and N. rossii suggest specific requirements in the mobilization, transport, incorporation, and/or catabolism of lipids that were reinforced by differences on some FA ratios expressing the activity coefficient of enzymes implicated on the FA pathway flux. [Copyright &y& Elsevier]

Abstract: Two chymotrypsins (chymotrypsin A and B) have been purified to homogeneity from the hepatopancreas of crucian carp (Carassius auratus) by ammonium sulphate fractionation and chromatographies on DEAE-Sepharose, Sephacryl S-200 HR, Phenyl-Sepharose and SP-Sepharose. The molecular masses of chymotrypsin A and B were approximately 28 and 27kDa, respectively, by SDS–PAGE. Purified chymotrypsins also revealed single bands by native-PAGE. Optimum temperatures of chymotrypsin A and B were 40 and 50°C, and optimal pHs were 7.5 and 8.0 using Suc-Leu-Leu-Val-Tyr-AMC as substrate. Both enzymes were effectively inhibited by serine proteinase inhibitors and slightly activated by metal ions such as Ca2+ and Mg2+, while inactivated by Mn2+, Cd2+, Cu2+, Fe2+ to different degrees. Apparent K ms of chymotrypsin A and B were 1.4 and 0.5μM, and K cats were 2.7S−1 and 3.4S−1, respectively. Immunoblotting analysis using anti-chymotrypsin B weakly cross reacted with chymotrypsin A. [Copyright &y& Elsevier]

Denaturing Gradient Gel Electrophoresis (DGGE) is a molecular fingerprinting method that separates similar size (bp), but different sequence of PCR products using acrylamide gel that contain denaturing gradient of urea and formamide. GC clamp is added at the 5′ end of one of the primer to prevent the PCR amplicon from completely denature. Different bacteria have different DNA sequences that will denature at different denaturant concentrations. Different DGGE conditions are suitable for different bacterial communities. This research aimed to optimize the conditions for determining bacterial community of fish intestinal tract. The percent of polyacrylamide gel (8.0% and 10.0%), denaturant concentrations (45 - 55%, 30 - 60%, and 30 - 70%), running times (5 h and 12 h) and voltages used (120 V and 200 V) were varied. The results indicated that, 8.0% polyacrylamide gel, 30 - 60% denaturant, 12 h running time and 120 V was the optimum condition for separation of different species of fish intestinal tract micro flora. [ABSTRACT FROM AUTHOR]

A 60-day feeding trial was carried out with juvenile Jian carp ( Cyprinus carpio var. Jian) to study the effects of myo-inositol (MI) on the growth, digestive enzyme and intestinal microbial population. Diets with seven levels of inositol (163.5, 232.7, 384.2, 535.8, 687.3, 838.8 and 990.3 mg MI kg−1 diet) were fed to Jian carp (initial weight 22.28±0.07 g). Per cent weight gain (PWG) was improved with increasing inositol levels up to 535.8 mg MI kg−1 diet ( P<0.05), and plateaued ( P>0.05). The protein production value, lipid production value and ash production value were increased with increasing dietary inositol levels up to 384.2, 838.8 and 838.8 mg MI kg−1 diet respectively ( P<0.05). Although intestinal protein content and trypsin activity were not affected by inositol levels ( P>0.05), chymotrypsin, lipase and amylase activities in intestine were the lowest for fish fed the MI-unsupplemented diet ( P<0.05). Alkaline phosphatase, Na+, K+-ATPase, γ-glutamyl transpeptidase and creatinkinase activities in the intestine were increased with an increase in the inositol levels up to 384.2–687.3 mg MI kg−1 diet ( P<0.05). Intestinal Aeromonas hydrophila and Escherichia coli decreased with an increase in the levels of dietary inositol up to 232.7 and 687.3 mg MI kg−1 diet respectively ( P<0.05), while Lactobacillus in the intestine increased with an increase in inositol levels up to 990.3 mg MI kg−1 diet ( P<0.05). In conclusion, inositol improved growth, digestive capacity and intestinal microbial population of juvenile Jian carp, and the dietary inositol requirement for PWG of juvenile Jian carp is 518.0 mg MI kg−1 diet. [ABSTRACT FROM AUTHOR]

Five iso-nitrogenous (300 g crude protein kg−1 diet) semi-purified diets with graded levels of carbohydrate at 220 (D-1), 260 (D-2), 300 (D-3), 340 (D-4) and 380 (D-5) g kg−1 diet were fed ad libitum to Puntius gonionotus fingerlings (average weight 0.59±0.01 g) in triplicate groups (20 fish replicate−1) for a period of 90 days to determine the effect of the dietary carbohydrate level on the growth, nutrient utilization, digestibility, gut enzyme activity and whole-body composition of fish. Fifteen flow-through cement tanks of 100 L capacity with a flow rate of 0.5 L min−1 were used for rearing the fish. The maximum weight gain, specific growth rate, protein efficiency ratio, RNA:DNA ratio, whole-body protein content, protease activity, protein and energy digestibility and minimum feed conversion ratio (FCR) were found in the D-2 group fed with 260 g carbohydrate kg−1 diet. The highest protein and energy retention was also recorded in the same group. However, from the second-order polynomial regression analysis, the maximum growth and nutrient utilization of P. gonionotus fingerlings was 291.3–298.3 g carbohydrate kg−1 diet at a dietary protein level of 300 g kg−1 with a protein/energy (P/E) ratio of 20.58 −20.75 g protein MJ−1. [ABSTRACT FROM AUTHOR]

1-Chloro-2,2-bis(4-chlorophenyl)ethene (p,p'-DDMU), a metabolite of either 1,1-dichloro-2,2-bis(4-chlorophenyl)ethane (p,p'-DDD) or 1,1-dichloro-2,2-bis(4-chlorophenyl)ethepe (p,p'-DDE), which is degraded from 1 ,11-trichloro-2,2-bis(4-chlorophenyl)ethane (p,p'-DDT), was detected in a variety of environmental matrices of the Pearl River Delta (PRD), South China, including fish, fish feeds, semidigested fish stomach contents, marine surface sediment, surface soil, atmospheric gaseous phase, and particulates, rainwater, and coastal rainwater. For fish species, the concentrations of p,p'-DDMU were significantly higher in farmed fish than in marine wild fish, with the highest value obtained in seawater farmed fish species (mean/median values of 262/173 ng/g lipid). The relative abundance of p,p'-DDMU to total DDTs (sum of o,p'- and p,p'-DDT, DDD, and DDE) was higher in samples of marine origin (~5%) than in those of terrestrial origin (~2%). Because p,p'-DDD was considerably abundant in all samples and a negative linear correlation was found between p,p'-DDD/(p,p'-DDD + p,p'-DDE) and p,p'-DDMU/DDTs in the marine sediments r2 = 0.73) and seawater farmed fish (r2 = 0.29) under investigation, it is possible that DDMU was partially dehydrochlorinated from DDD in the environment of the PRD. A fugacity-based assessment suggested that p,p'-DDMU is likely to transport from sediment to seawater and then to air and from soil to air. The ubiquity of p,p'- DDMU in the PRD indicates that it may also be widespread worldwide particularly in countries with large amounts of DDT used currently and/or historically. Given the potential risk of p,p'- DDMU to human health, more efforts should be directed toward to this previously overlooked contaminant. [ABSTRACT FROM AUTHOR]