Alessio Bonaldo

Exploring the larval transcriptome of the common sole ( Solea solea L.)

BackgroundThe common sole (Solea solea) is a promising candidate for European aquaculture; however, the limited knowledge of the physiological mechanisms underlying larval development in this species has hampered the establishment of successful flatfish aquaculture. Although the fact that genomic tools and resources are available for some flatfish species, common sole genomics remains a mostly unexplored field. Here, we report, for the first time, the sequencing and characterisation of the transcriptome of S. solea and its application for the study of molecular mechanisms underlying physiological and morphological changes during larval-to-juvenile transition.ResultsThe S. solea transcriptome was generated from whole larvae and adult tissues using the Roche 454 platform. The assembly process produced a set of 22,223 Isotigs with an average size of 726 nt, 29 contigs and a total of 203,692 singletons. Of the assembled sequences, 75.2% were annotated with at least one known transcript/protein; these transcripts were then used to develop a custom oligo-DNA microarray. A total of 14,674 oligonucleotide probes (60 nt), representing 12,836 transcripts, were in situ synthesised onto the array using Agilent non-contact ink-jet technology. The microarray platform was used to investigate the gene expression profiles of sole larvae from hatching to the juvenile form. Genes involved in the ontogenesis of the visual system are up-regulated during the early stages of larval development, while muscle development and anaerobic energy pathways increase in expression over time. The gene expression profiles of key transcripts of the thyroid hormones (TH) cascade and the temporal regulation of the GH/IGF1 (growth hormone/insulin-like growth factor I) system suggest a pivotal role of these pathways in fish growth and initiation of metamorphosis. Pre-metamorphic larvae display a distinctive transcriptomic landscape compared to previous and later stages. Our findings highlighted the up-regulation of gene pathways involved in the development of the gastrointestinal system as well as biological processes related to folic acid and retinol metabolism. Additional evidence led to the formation of the hypothesis that molecular mechanisms of cell motility and ECM adhesion may play a role in tissue rearrangement during common sole metamorphosis.ConclusionsNext-generation sequencing provided a good representation of the sole transcriptome, and the combination of different approaches led to the annotation of a high number of transcripts. The construction of a microarray platform for the characterisation of the larval sole transcriptome permitted the definition of the main processes involved in organogenesis and larval growth.

The influence of dietary β-glucans on the adaptive and innate immune responses of European sea bass (Dicentrarchus labrax) vaccinated against vibriosis

Abstract The effects of feeding 1,3/1,6 β-glucans on the innate and the adaptive immune responses of European sea bass (Dicentrarchus labrax) was investigated. Two experiments were carried out during the study. In the first, a number of non-specific immune parameters were examined at 4, 7, 10, 14, 21 and 25 days of feeding fish with a semipurified diet containing Macrogard ©, a commercially available form of 1,3/1,6 β-glucans. The respiratory burst activity of head kidney macrophages isolated from the different groups of fish fed the immunostimulant peaked and subsequently decreased at different times during the experiment. Head kidney macrophages from fish fed 250 ppm β-glucans had a statistically higher level of respiratory burst activity at Day 21 of feeding compared with fish fed no immunostimulant. No statistical differences were observed in lyzozyme activity during this trial. In the second experiment, the effect of feeding 1,3/1,6 β-glucans on the immune response of fish to an alginate-encapsulated Vibrio vaccine administered orally was examined. Respiratory burst of head kidney macrophages and serum lysozyme activity decreased in all fish over the course of the trial, while serum lysozyme activity was considerably lower than values obtained in the first experiment. Fish vaccinated orally had significant increases in antibody response by Week 2 post-vaccination, but β-glucans did not appear to affect these levels. Vaccination may have resulted in activating the immune system as a whole, thus masking any difference in immunostimulation by the β-glucans. It may be that the optimal doses and timing of β-glucans administration is different when the immunostimulant is administered alone or in combination with the vaccine. In conclusion, European sea bass can be immunomodulated with oral administration of β-glucan. Optimal doses and administration times have been established when β-glucans are fed alone, although further studies are needed to establish ideal feeding regimes for glucans administrated in combination with vaccination.

Effect of feeding a selected combination of galacto-oligosaccharides and a strain of Bifidobacterium pseudocatenulatum on the intestinal microbiota of cats

OBJECTIVE To evaluate the growth kinetics of a strain of Bifidobacterium pseudocatenulatum (BP) on 4 oligo- or polysaccharides and the effect of feeding a selected probiotic-prebiotic combination on intestinal microbiota in cats. ANIMALS 10 healthy adult cats. PROCEDURES Growth kinetics of a strain of cat-origin BP (BP-B82) on fructo-oligosaccharides, galacto-oligosaccharides (GOS), lactitol, or pectins was determined, and the combination of GOS and BP-B82 was selected. Cats received supplemental once-daily feeding of 1% GOS-BP-B82 (10(10) CFUs/d) for 15 days; fecal samples were collected for analysis the day before (day 0) and 1 and 10 days after the feeding period (day 16 and 25, respectively). RESULTS Compared with the prefeeding value, mean fecal ammonia concentration was significantly lower on days 16 and 25 (288 and 281 μmol/g of fecal dry matter [fDM], respectively, vs 353 μmol/g of fDM); fecal acetic acid concentration was higher on day 16 (171 μmol/g of fDM vs 132 μmol/g of fDM). On day 16, fecal concentrations of lactic, n-valeric, and isovaleric acids (3.61, 1.52, and 3.55 μmol/g of fDM, respectively) were significantly lower than on days 0 (5.08, 18.4, and 6.48 μmol/g of fDM, respectively) and 25 (4.24, 17.3, and 6.17 μmol/g of fDM, respectively). A significant increase in fecal bifidobacteria content was observed on days 16 and 25 (7.98 and 7.52 log(10) CFUs/g of fDM, respectively), compared with the prefeeding value (5.63 log(10) CFUs/g of fDM). CONCLUSIONS AND CLINICAL RELEVANCE Results suggested that feeding 1% GOS-BP-B82 combination had some positive effects on the intestinal microbiota in cats.

Effects of dietary lipid level on growth and feed utilisation of gilthead seabream ( Sparus aurata L.) reared at Mediterranean summer temperature

We investigated the effects of different dietary lipid levels on gilthead seabream, Sparus aurata, reared at Mediterranean summer temperature. Sixty fish (average weight 75 g) per tank were randomly distributed, in triplicate groups, in a recirculating rearing system (27±1°C) and fed ad libitum five isonitrogenous (46% dietary protein) diets with increasing lipid level (16, 18, 20, 22 and 24% named D16, D18, D20, D22 and D24, respectively), over 89 days. Specific growth rate and final body weight were not affected by dietary lipid levels. Feed conversion ratio was significantly higher (P≤0.05) in D16 as compared to the other treatments, most likely due to the shortage of dietary energy supply, coped with a significantly higher voluntary feed intake. Consequently, we obtained a significantly lower protein efficiency ratio and gross protein efficiency in D16. Gross lipid efficiency was significantly higher in D16 and D18 than in the other treatments. Biometric parameters and lipase activity in gut content were not influenced by dietary treatments. In conclusion, D18 seems the most suitable diet for gilthead seabream reared at Mediterranean summer temperature, providing both the lowest fish in fish out (FIFO) ratio and a protein sparing effect, which makes gilthead seabream’s production economically and environmentally more sustainable.

Fatty acid composition of eggs and its relationships to egg and larval viability from domesticated common sole (Solea solea) breeders.

The study of lipids and fatty acids (FAs) has been used in the assessment of egg quality because their composition can influence the fertilization rate, hatching, survival and growth of marine fish larvae. For these reasons, the lipid content (TL) and fatty acid composition of common sole (Solea solea) eggs were measured and correlated to egg and larval viability parameters throughout an entire reproductive season. Seventeen batches of fertile eggs obtained from natural spawning of captive breeders were characterized for the TL, FA profile, hatching rate (HR) and survival rate of larvae (SR) at 0-6 days post-hatching (dph). The egg FA composition reflected the composition of the feed supplied to the broodstock during summer and autumn (before and during vitellogenesis) rather than that supplied during the spawning season. In general, the egg FA profile showed minimal differences among the early-, mid- and late-spawning periods (possibly due to the change of the diet and/or water temperature) indicating that it is possible to obtain a similar egg quality in terms of egg FA profile over 2 months of spawning. Saturated FAs and monounsaturated FAs (MUFA) were positively correlated with HR, while TL, 22 : 6n-3 (DHA), 20 : 4n-6 (ARA), polyunsaturated FAs of the (n-3) series (n-3 PUFA) and polyunsaturated FAs of the (n-6) series were negatively correlated (p ≤ 0.05). MUFA, 20 : 5n-3 (EPA), n-6/n-3 were positively correlated with SR, while DHA, n-3 PUFA, DHA/EPA were negatively correlated (p ≤ 0.05). In conclusion, the feed supplied before and during vitellogenesis has a major role in determining the egg FA profile in common sole. The relationships found between TL and FAs with egg and larval viability parameters differ from many other farmed marine fish species, which may suggest the need for a specific broodstock feed for this species.

Use of centrifuged and preserved microalgae for feeding juvenile Manila clam (Tapes philippinarum): effects on growth and fatty acid composition

Abstract The objective of the present study was to assess the nutritional value of three diets containing commercial preserved microalgae pastes and the relationship of their biochemical composition to the growth rate and fatty acids profile of juvenile Tapes philippinarum. The feeding period was 6 weeks. At a daily ratio of 1% dry weight live weight of the clams–1, the trispecific diet with 40% of I. galbana, 20% of T. suecica and 40% of C. muelleri supported greater growth of animals than either a mixture of 80% of I. galbana and 20% T. suecica or I. galbana alone (P< 0.05). Even the fatty acid profile was significantly modified depending on the composition of the diets. In particular a decrease of EPA was observed in clams with the lowest growth rate.

Growth response and body composition of sharpsnout sea bream (Diplodus puntazzo) fed a high energy diet with different protein levels

Abstract A study was undertaken to determine the effect of a high energy diet with two different protein levels on growth, feed efficiency and whole body composition of sharpsnout sea bream (Diplodus puntazzo). Two isoenergetic diets (24.1–24.7 MJ Kg-1 dry weight) with two different protein levels (46.7 and 52.5% dry weight) were fed to satiety to duplicate groups of 300 fish (initial body weight 27.7 ± 0.2 g) for 94 days. At the end of the experiment, the fish fed 52.5% protein showed a statistically higher (P < 0.05) daily intake rate (DIR) of feed. Feed conversion rate (FCR) was similar among groups. Whole body composition was similar among treatments while the high energy level of the diets significantly modified lipid and moisture content in comparison with fish at the beginning of the experiment. Protein efficiency ratio (PER) and gross protein efficiency (GPE) of fish fed 46.7% protein diet were statistically higher than those for the other diet. It may be concluded that the diet with a lower protein level has given better protein utilization and a protein sparing effect but tended to result in reduced weight gain and feed intake, when compared with diet containing higher protein levels.

Growth parameters, behavior, and meat and ham quality of heavy pigs subjected to photoperiods of different duration

To attain a good level of animal welfare, pigs require a sufficient environmental illumination. Therefore, minimum levels for light duration and light intensity have been set up by the European legislation (Directive 2008/120). An experimental trial was designed to determine whether an increased duration of the photophase (up to 16 h of light per day) could modify the behavior, productive parameters, and meat and ham quality of Italian heavy pigs. Forty crossbred (Large White × Landrace) castrated males pigs (26 kg initial average BW) intended for Protected Designation of Origin (according to European Union Regulation 1151/2012, ) dry-cured ham production were raised according to Parma ham production rules up to the weight of 160 kg. Pigs were homogeneously allotted to 2 experimental groups, each comprising 20 pigs. The short photoperiod (SP) group received the minimum mandatory number of hours of light per day (corresponding to 8 h/d), whereas the long photoperiod (LP) group was subjected 16 h of light per day during the whole production cycle. Light intensity was maintained at 40 lux (i.e., the minimum mandatory level) for both the experimental groups. Growth and slaughtering parameters, carcass traits, fatty acid composition, meat and dry-cured ham quality, and animal behavior were assessed. Pigs in the LP group showed a greater live weight and carcass weight compared to the SP group ( = 0.005 and = 0.007, respectively). Similarly, hams obtained from the LP group were significantly heavier and their weight losses during the dry-curing period were reduced ( < 0.01) when compared to the SP group. No significant differences were detected between the experimental groups as concerns meat and ham quality or fatty acid composition of the subcutaneous fat. Pigs in the LP group spent more time resting and less time pseudo-rooting ( < 0.01). Our results indicate that, given an appropriate dark period for animal rest, an increased duration of the photoperiod, even at the lower mandatory light intensity level, can favorably affect growth parameters of heavy pigs without any negative effect on animal behavior, carcass traits, or meat or long-cured ham quality. Therefore, rearing pigs in semidarkness should be considered as a baseless practice, contrary to animal welfare.

Effects of feeding low fishmeal diets with increasing soybean meal levels on growth, gut histology and plasma biochemistry of sea bass

The aquaculture industry depends upon the development of sustainable protein sources to replace fishmeal (FM) in aquafeeds and the products derived from soybeans are some of the most studied plant feedstuffs. A key area of investigation for continuing to improve modern aquafeeds includes the evaluation of varying proportions and combinations of plant ingredients to identify mixtures that are more efficiently utilized by the fish. This study investigated the effects of increasing soybean meal (SBM) by replacing a mix of plant ingredients in low FM (20%) diets on growth, blood biochemistry profile and gut histology on European sea bass. Five isonitrogenous and isolipidic experimental diets were formulated: four diets containing increasing SBM levels (0, 10, 20 and 30%; 0SBM, 10SBM, 20SBM and 30SBM, respectively) with a low content of FM (20%) and one control diet (0% SBM; 35% FM). Diets containing SBM brought to comparable performance and protein utilization, while 0SBM had negative impact on feed conversion rate and protein utilization. Blood parameters suggested an optimal nutritional status under all feeding treatments, even though slightly decreased values were reported at increasing dietary SBM. Histology examination did not show any changes indicative of soy-induced enteritis. We can conclude that for European sea bass: (i) different blends of plant protein did not affect feed intake despite the 20% FM dietary level; (ii) the inclusion of SBM maintains optimal growth and feed utilization in low FM diets; (iii) blood biochemistry profile showed a good nutritional status under all feeding regimes; (iv) no evidence of soy-induced enteritis was reported in any group fed low FM diets. For formulation of practical diets in on-growing of European sea bass, SBM up to 30% can be successfully incorporated into feeds containing low FM inclusion.

α-Transducin and α-gustducin immunoreactive cells in the stomach of common sole (Solea solea) fed with mussel meal.

Vertebrates perceive a variety of exogenous substances using two main chemosensory systems, taste and olfaction. The taste perception occurs through the interaction of taste receptors associated with specific G protein subunits such as α-transducin (Gαtran) and α-gustducin (Gαgust). Aquatic vertebrates are also provided with a chemosensory system consisting of solitary chemosensory cells distributed to the oropharynx and skin. In this study, we identified Gαtran and Gαgust-immunoreactive cells intermingled with non-labeled epithelial cells in the gastric mucosa of the common sole. A long-term diet with increasing concentrations of mussel meal in the protein component of a conventional fish meal-based diet induced a dose-dependent increase in the gastric epithelial area and density of Gαtran and Gαgust immunoreactive cells. These findings suggest that taste-related molecules are regulated by changes in diet formulation in common sole aquaculture.