Eleni Mente

White muscle free amino acid concentrations following feeding a maize gluten dietary protein in Atlantic salmon (Salmo salar L.)

Abstract This study aimed to investigate the effect of high maize gluten protein diets on the white muscle free amino acid (FAA) concentrations of Atlantic salmon following feeding and to examine whether these tissue free amino acids profiles highlighted any dietary deficiencies in essential amino acids (EAA). Two isocaloric and isonitrogenous diets (52% protein, 20% oil) were formulated and mixed proportionately to give a total of five feeds in which maize gluten provided 0%, 16%, 32%, 48% and 64% of the total dietary protein (diets 0MG, 16MG, 32MG, 48MG, 64MG, respectively). Triplicate groups of 19 g Atlantic salmon were fed each of the experimental diets for a period of 40 days. At the end of the growth experiment, groups of four salmon from each diet were taken at 4, 8 and 12 h after feeding for the white muscle amino acids analysis. Although fish doubled their weights during the growth period, no significant differences were observed in specific growth rates, feed conversion ratios, protein digestibility and protein growth rates among fish fed the test diets. Total free amino acid (FAA) concentrations did not change at various times after a meal and this was also true for the majority of individual FAAs (except asparagine, which increased significantly 12 h after feeding). The most notable diet-induced changes in the white muscle were an overall reduction in essential FAA concentrations (threonine and lysine) as dietary maize gluten content increased and an increase in histidine and leucine. Correlations between dietary essential amino acids patterns and white muscle protein-bound amino acids were examined. The possibility of partial replacement of fish meal maize gluten up to 50% was demonstrated.

Temporal shifts of the Norway lobster (Nephrops norvegicus) gut bacterial communities.

The aim of this study was to investigate the gut bacterial communities of Nephrops norvegicus individuals, using a suite of molecular tools consisting of automated ribosomal intergenic spacer analysis, 16S rRNA gene-internal transcribed spacer clone libraries and FISH. The animals were collected from Pagasitikos Gulf, Greece, during different months of the year. The diversity of the gut bacterial communities was found to mostly vary with sampling time, which could be related to temporal variations in food supply. The 16S rRNA gene diversity analysis showed dominance of specific phylotypes for each month studied. February, May, July, August and October samples were rich in sequences related to the gammaproteobacterial genera Pseudoalteromonas, Psychrobacter and Photobacterium. September and December samples were dominated by phylotypes affiliated with uncultured representatives of Mollicutes, which are generally associated with the intestinal tracts of various animals. The presence of Gammaproteobacteria and uncultured Mollicutes in August and September samples, respectively, was further confirmed by FISH. None of the morphometric parameters considered was related to the temporal pattern of dominant bacterial communities.

Dietary differences are reflected on the gut prokaryotic community structure of wild and commercially reared sea bream (Sparus aurata).

We compared the gut prokaryotic communities in wild, organically‐, and conventionally reared sea bream (Sparus aurata) individuals. Gut microbial communities were identified using tag pyrosequencing of the 16S rRNA genes. There were distinct prokaryotic communities in the three different fish nutritional treatments, with the bacteria dominating over the Archaea. Most of the Bacteria belonged to the Proteobacteria, Firmicutes, Actinobacteria, and Bacteroidetes. The number of bacterial operational taxonomic units (OTUs) was reduced from the wild to the conventionally reared fish, implying a response of the gut microorganisms to the supplied food and possibly alterations in food assimilation. The dominant bacterial OTU in all examined fish was closely related to the genus Diaphorobacter. This is the first time that a member of the β‐Proteobacteria, which dominate in freshwaters, are so important in a marine fish gut. In total the majority of the few Archaea OTUs found, were related to methane metabolism. The inferred physiological roles of the dominant prokaryotes are related to the metabolism of carbohydrates and nitrogenous compounds. This study showed the responsive feature of the sea bream gut prokaryotic communities to their diets and also the differences of the conventional in comparison to the organic and wild sea bream gut microbiota.

Gut bacteria associated with different diets in reared Nephrops norvegicus

The impact of different diets on the gut microbiota of reared Nephrops norvegicus was investigated based on bacterial 16S rRNA gene diversity. Specimens were collected from Pagasitikos Gulf (Greece) and kept in experimental rearing tanks, under in situ conditions, for 6months. Treatments included three diets: frozen natural (mussel) food (M), dry formulated pellet (P) and starvation (S). Gut samples were collected at the initiation of the experiment, and after 3 and 6months. Tank water and diet samples were also analyzed for bacterial 16S rRNA gene diversity. Statistical analysis separated the two groups fed or starved (M and P vs. S samples). Most gut bacteria were not related to the water or diet bacteria, while bacterial diversity was higher in the starvation samples. M and P samples were dominated by Gammaproteobacteria, Epsilonproteobacteria and Tenericutes. Phylotypes clustering in Photobacterium leiognathi, Shewanella sp. and Entomoplasmatales had high frequencies in the M and P samples but low sequence frequencies in S samples. The study showed that feeding resulted in the selection of specific species, which also occurs in the natural population, and might be associated with the animals nutrition.

The presence of microcystins in fish Cyprinus carpio tissues: a histopathological study

The occurrence of heavy cyanobacterial blooms has become a worldwide problem, as a consequence of eutrophication of the aquatic ecosystems; furthermore, 60% to 75% of these blooms have been found to be toxic. Microcystins (MCYSTs), the predominant toxins of cyanobacterial blooms, are associated with mortality and illness in both animals and humans. Laboratory-controlled experiments studying the effects of different microcystins on the common carp (Cyprinus carpio) have revealed various histopathological alterations. The aim of the present study is to investigate the effect of chronic or subchronic exposure of fish to microcystins under natural environmental conditions by examining the possible histopathological changes associated with a dense cyanobacterial bloom and determining the microcystin contents of fish tissues. Common carps (C. carpio) were caught from Lake Karla (Greece), during a dense cyanobacterial bloom. The concentration of MCYSTs in the fish liver, kidney and muscle tissues was measured by enzyme-linked immunosorbent assay. The pseudogaster contents were analysed, and a histopathological examination was performed using light and electron microscopy. Severe alterations were detected in the liver and the kidney, suggesting that the toxic effects were caused by various pollutants that were particularly associated with microcystins. The histopathological findings are also discussed, taking into consideration the health conditions of the common carp as a commercial fish species. The mechanisms of expansion of the microcystins and the poisoning of aquatic organisms (e.g. fish) are not yet known in the Lake Karla ecosystem. Future research may focus on identifying the changes caused by microcystins and other factors that exert similar effects on fish tissues, as well as on establishing the overall combined effect of all these factors on fish health.

Effects of Different Diets on the Digestive Physiology of Adult Norway Lobster Nephrops norvegicus

ABSTRACT Rearing Norway lobsters for restocking is a management option to compensate fishing overexploitation, but this requires, among other topics, developing specific pellet diets. The activity of digestive enzymes and the histological organization of the hepatopancreas were analyzed to assess the effectiveness of a pellet diet on digestive performance. Thirty animals were reared individually for 6 mo, fed with mussels and pellets or starved. Comparisons with wild lobsters were also conducted. Survival rate of lobsters fed with frozen mussels was the greatest. Trypsin and amylase activity in the hepatopancreas of the lobsters fed pellets was greater than that of other treatments. In contrast, pepsin activity in the gastric juice of starved animals was the greatest. Levels of alkaline proteases in the gastric fluid were greater in lobsters fed with mussels and the pellet diet in contrast to fasted animals. Amylase was greater in specimens fed frozen mussels. The greatest level of lipid accumulation observed in lobsters fed with the pellet diet indicated a physiological imbalance between the absorptive and transport capacity of the hepatopancreatic cells resulting from an excess of dietary lipids. These results might serve as a reference for future studies for formulating feeds specifically designed for Nephrops norvegicus.

A novel image processing method to determine the nutritional condition of lobsters

The digestive gland of crustacean is involved in various metabolic activities, including the synthesis and secretion of digestive enzymes that begin the process of food digestion, intracellular digestion and absorption of nutrients, storage of reserves, and disposal of waste products. It consists of two glandular lobes which extensively subdivide to form a complex of blind-ending tubules, whose size, surface area, and digestive cells are associated with intracellular digestion and the nutritional status of the organism. The aim of this paper was to study the morphology of the digestive gland in various lobster species and calculate the surface area of tubules, lumen and digestive cells (R-, F-, and B-cells) and their ratios to total tubule surface area. The similarity in ratios obtained in this study between individual lobsters suggests that the method developed in this study can be successfully applied to a range of species. This study describes a novel image processing algorithm for the automatic measurement of the hepatopancreas structure using stained cross sections of digestive gland tubules. The proposed new methodology could be used for studying the physiology and nutrient metabolism of lobsters and other crustaceans. The computer-aided analysis described in this paper is accurate for the quantitative assessment of the lobsters digestive gland structure.

Changes of bacterioplankton apparent species richness in two ornamental fish aquaria

We analysed the 16S rRNA gene diversity within the bacterioplankton community in the water column of the ornamental fish Pterophyllum scalare and Archocentrus nigrofasciatus aquaria during a 60-day growth experiment in order to detect any dominant bacterial species and their possible association with the rearing organisms. The basic physical and chemical parameters remained stable but the bacterial community at 0, 30 and 60 days showed marked differences in bacterial cell abundance and diversity. We found high species richness but no dominant phylotypes were detected. Only few of the phylotypes were found in more than one time point per treatment and always with low relative abundance. The majority of the common phylotypes belonged to the Proteobacteria phylum and were closely related to Acinetobacter junii, Pseudomonas sp., Nevskia ramosa, Vogesella perlucida, Chitinomonas taiwanensis, Acidovorax sp., Pelomonas saccharophila and the rest belonged to the α-Proteobacteria, Bacteroidetes, Actinobacteria, candidate division OP11 and one unaffiliated group. Several of these phylotypes were closely related to known taxa including Sphingopyxis chilensis, Flexibacter aurantiacus subsp. excathedrus and Mycobacterium sp. Despite the high phylogenetic diversity most of the inferred ecophysiological roles of the found phylotypes are related to nitrogen metabolism, a key process for fish aquaria.

Shift in trophic level of Mediterranean mariculture species.

The mean trophic level of the farmed fish species in the Mediterranean has been increasing. We examined the farming-up hypothesis (i.e., the increase in the production of high-trophic-level species) in the Mediterranean by determining the trophic level of the aquafeeds (i.e., what the fish are fed) of 5 species of farmed marine fishes: common dentex (Dentex dentex), common pandora (Pagellus erythrinus), European seabass (Dicentrarchus labrax), gilthead seabream (Sparus aurata), and red porgy (Pagrus sp.). The mean trophic level of aquafeed used in mariculture from 1950 to 2011 was higher (3.93) than the prey farmed fish consume in the wild (3.72) and increased at a faster rate (0.48/decade) compared with that based on their diets in the wild (0.43/decade). Future expected replacement of the fishmeal and oil in aquafeeds by plant materials may reverse the farming-up trend, although there are a number of concerns regarding operational, nutritional, environmental, and economic issues. The farming-up reversal can be achieved in an ecologically friendly manner by facilitating the mariculture of low-trophic-level fishes and by promoting high efficiency in the use of living marine resources in aquafeeds.

Collagen Fibrils in Cultured and Wild Sea Bream (Sparus aurata) Liver. An Electron Microscopy and Image Analysis Study

This study aims to measure liver collagen fibril diameter in cultured and wild sea breams (Sparus aurata). Cultured sea breams were fed three isonitrogenous diets. The organically produced feed contained sustainable certified fish meal (45%), fish oil (14%), and organic certified wheat; the laboratory feed contained fish meal (45%), fish oil (14%), wheat meal, and soya meal; and the commercial feed included fish meal (46%), fish oil (17%), soya meal, wheat meal, and corn gluten meal. The organic diet had higher amounts of vitamins A, C, and E; specific amino acids; and minerals that enhanced the biosynthesis of collagen. This study shows that fish fed the organic feed had significantly bigger collagen fibril diameters than the fish fed the conventional feed. Furthermore, the organically fed fish had similarly sized collagen fibril diameters as wild fish. More research is needed to understand the long-term effects and the mechanism and function of fish collagen peptide intake on lipid absorption and metabolism; and to identify dietary regimes that are able to improve whole body lipid profiles and suppress the transient increase of plasma triglycerides.