Fish Physiology and Biochemistry最新文献

To investigate the influences of dietary protein and lipid levels on the growth, feed utilization, morphometric parameters, body composition, serum biochemical parameters, and lipid metabolism of golden pompano (Trachinotus ovatus), nine test diets containing three protein levels (35%, 40%, and 45%) and three lipid levels (8%, 13%, and 18%) were designed in the present study. Each diet (named D1-D9) was randomly assigned to feed triplicate groups of golden pompano juvenile (initial weight ~ 70 g) for 50 days. The results showed that the dietary lipid levels positively correlated with weight gain, specific growth rate, and protein efficiency ratio (PER), suggesting that the high lipid diets (18%) can be efficiently utilized in this fish species. The dietary protein levels have no significant influences on the growth and feed utilization except for the PER. Increasing dietary protein levels resulted in a decrease in hepatosomatic index (HSI), viscerosomatic index (VSI), and intestinal somatic index (ISI), while the dietary lipid level did not have a significant impact on morphological indices except for ISI. The dietary protein and lipid levels had no significant influences on the contents of crude lipid, crude ash, and moisture of whole body, while the crude protein contents was significantly affected by the dietary protein levels. Serum biochemical indexes, including cholesterol (CHO), triglycerides (TG), high-density lipoprotein cholesterol (HDL), and low-density lipoprotein cholesterol (LDL), as well as HDL/LDL ratio were significantly affected by the dietary lipid levels, but not by the dietary protein levels. The expression levels of genes and their associated proteins involved in hepatic lipogenesis (Srebp-1c and Fas) and fatty acids β-oxidation (Pparα and Cpt-1) were up-regulated with increasing dietary lipid levels, while the former was up-regulated, and the latter was down-regulated with increasing dietary protein levels. Considering the present results in terms of growth performance, feed utilization, morphometric parameters, and lipid metabolism, the recommended dietary protein and lipid levels for golden pompano are 40% and 18%, respectively. The findings suggested that this species exhibits a significant protein-sparing effect on lipid utilization.

Chirostoma estor (Jordan, 1879) is an endemic freshwater species with a high potential for aquaculture; however, as in many other fish, larviculture of this species is the most critical stage, in which the higher mortality rates. Therefore, it is necessary to fully describe the development of the digestive system to establish better feeding protocols in the larval culture of C. estor, both for aquaculture and restoration purposes. In the present study, larviculture was carried out from hatching to 20 days after hatching (DAH). The organisms were fed with the rotifer Brachionus plicatilis from 2 to 14 DAH, and nauplii of Artemia sp. from 15 to 20 DAH. A total of 12 organisms (0, 3, 5, 10, 15, and 20 DAH) were taken for size and weight growth and histological and histochemical analysis. The histological analysis indicated that after 3 DAH, the opening of the mouth and anus was observed, coinciding with the beginning of exogenous feeding. In addition, the digestive system developed, with differentiation of the oropharyngeal sections, esophagus, and intestine, folding of the intestinal mucosa, as well as associated organs (liver and pancreas) that reach their maximum development at 20 DAH. Thus, C. estor at this stage of development can digest and absorb nutrients despite being an agastric fish. The results obtained in this study will facilitate a better understanding of the ontogenetic morphophysiological development processes, associated with the transition of larvae to exogenous feeding, which ensures a higher percentage of survival during larval development and of course, adds to the diversity ontogenetics of teleostean.

To induce sexual maturation in captivity, eels rely on hormonal treatments, but this process is costly and time-consuming. As an alternative, different types of conditioning, also referred as pre-treatment, have been assessed to ease hormonal treatment response. Recent studies have shown that migrating eels experience a wide range of temperatures, varying from 12 °C at night to as low as to 8 °C during the day. Therefore, this study evaluates the effects of low-temperature (10 °C) seawater pre-treatments of different durations (2 and 4 weeks) on male eel reproduction. The eye, gonadosomatic and hepatosomatic indexes from control (without thermic seawater pre-treatment) and pre-treated fish were measured. Blood and testis samples were also collected for sex steroid and histology analysis, respectively. Eels pre-treated for 2 weeks demonstrated increased progestin levels, comparing with the control group. Eels pre-treated for 4 weeks showed significantly higher gonadosomatic index and elevated androgens and estradiol levels in comparison with the remaining groups. In eels pre-treated for 2 and 4 weeks, there was an increase in the proportion of spermatogonia type B cells compared to undifferentiated spermatogonia type A, a differentiation process that was not observed in the control group. Cold seawater pre-treatment induced early sexual maturation, including steroid production, which consequently stimulated biometric changes and increased spermatogonia differentiation. Following the pre-treatments, eels started receiving standard hormonal treatment (with recombinant human chorionic gonadotropin at 20 °C). Pre-treated males started to spermiate earlier than the control group. In some treatment weeks, pre-treated individuals registered higher values of sperm density, motility, and kinetic parameters. Moreover, an economic evaluation was carried out relating the investment made in terms of hormone injections with the volume of high-quality sperm obtained from each experimental group. The low temperature pre-treatments demonstrated their economic effectiveness in terms of hormone treatment profitability, increasing the production of high-quality sperm in the European eel. Thus, this in vivo study suggests that cold seawater pre-treatment may increase sensitivity to the hormone applied during standard maturation treatment.

This study focuses in investigating the fatty acid contents of surviving infected hybrid grouper fed with oleic acid immunostimulant. After a 6-week feeding trial, Epinephelus fuscoguttatus × Epinephelus lanceolatus fingerlings were infected with Vibrio vulnificus. One week after bacterial challenge, fish oil was extracted from body tissue of surviving infected fingerlings using the Soxhlet extraction method. The extracted samples were then sent for GC-MS analysis. The raw GC-MS data were analyzed using software programs and databases (i.e., MetaboAnalyst, SIMCA-P, NIST Library, and KEGG). A total of 39 metabolites were putatively identified, with 18 metabolites derived from the fatty acid group. Our further analysis revealed that most metabolites were highly abundant in the oleic acid dietary samples, including oleic acid (4.56%), 5,8,11-eicosatrienoic acid (3.45%), n-hexadecenoic acid (3.34%), cis-erucic acid (2.76%), and 9-octadecenoic acid (2.5%). Worthy of note, we observed a greater abundance of α-linoleic acid (15.57%) in the control diet samples than in the oleic acid diet samples (14.59%) with no significant difference in their results. The results obtained from this study revealed that surviving infected hybrid grouper expressed more immune-related fatty acids due to the effect of oleic acid immunostimulant. Therefore, in this study, we propose oleic acid as a potential immunostimulant in enhancing fish immunity in aquaculture industry.

In this study, the full-length cDNA sequences of the phosphatidylinositol-3-kinase p85 alpha (PI3KR1) and serine/threonine kinase 1 (AKT1) genes in largemouth bass (Micropterus salmoides) were obtained using the rapid amplification of cDNA ends (RACE) method. Sequence analysis revealed that the cloned sequences of PI3KR1 and AKT1 are 4170 bp and 3672 bp in length, with open reading frames (ORFs) of 1389 bp and 1422 bp encoding 462 and 473 amino acids, respectively. Sequence alignment and evolutionary tree analysis indicated their close relationship to other teleosts, especially those with similar feeding habits. Tissue distribution demonstrated widespread distribution of both genes in various tissues, with the highest abundance in the liver. Further results found that the upregulation of the expression of p-PI3KR1, p-AKT1, p-FoxO1, and GLUT2 proteins by insulin, while suppressing the expression of the total FoxO1 protein, effectively triggers a significant activation of the PI3KR1-AKT1 insulin signaling pathway. Meanwhile, the mRNA levels of the key glycolytic genes, including glucokinase (gk), pyruvate kinase (pk), and phosphofructokinase liver type (pfkl), have been enhanced evidently. In contrast, the expression of gluconeogenic genes such as phosphoenolpyruvate carboxykinase (pepck), glucose-6-phosphatase catalytic subunit (g6pc), and fructose-1,6-bisphosphatase-1 (fbp1) has been notably down-regulated. In addition, insulin treatment promoted the phosphorylation of glycogen phosphorylase (PYGL) and the dephosphorylation of glycogen synthase (GS), and the glycogen content in the insulin-treated group was remarkably reduced compared to the control group. Overall, our study indicates that the activation of PI3KR1-AKT1 insulin signaling pathway represses the hepatic glycogen deposition via the regulation of glycolysis and gluconeogenesis, which provides some new insights into nutritional strategy to effectively regulate the glucose metabolism in carnivorous fish.

This study aimed to investigate the effects of dietary metformin supplementation on the redox balance, inflammation, mitochondrial biogenesis, and function in blunt snout bream fed a high-carbohydrate (HC) diet. Fish (45.12 ± 0.36 g) were randomly offered four diets, including a control diet (33% carbohydrate), an HC diet (45% carbohydrate), and the HC diet supplemented with 0.06% (HCM1) and 0.12% (HCM2) metformin respectively for 12 weeks. Compared with the control, feeding the HC diet significantly increased the hepatosomatic index (HSI), the mesenteric fat index, liver and muscle glycogen contents, liver and adipose tissue lipid contents, plasma glucose and glycation end products (AGES) levels and aspartate transaminase activity, plasma and liver malondialdehyde (MDA) contents, hepatic adenosine triphosphate (ATP) and adenosine monophosphate (AMP) contents, mitochondrial cytochrome c content, mitochondrial complex IV activity and ATP 6 transcription, but decreased plasma catalase (CAT) activity, muscle superoxide dismutase (SOD) activity, hepatic antioxidant enzymes activities, and the transcriptions of transforming growth factor β (tgfβ) and interleukin 10 (il10). Compared with the HC group, metformin treatment (especially the HCM2 group) significantly elevated tissue glycogen contents, muscle SOD activity, plasma and liver antioxidant enzymes activities, the transcriptions of tgfβ and il10, the sodium/potassium ATPase activity, the contents of mitochondrial protein and AMP, the level of p-AMP activated protein kinase (AMPK), and the p-AMPK/t-AMPK ratio, but lowered the HSI, tissue lipid contents, plasma levels of glucose, AGES and glycated serum protein, plasma, and liver MDA contents, the transcriptions of il1β, NADH dehydrogenase subunit 1 and ATP 6, the contents of ATP and cytochrome c, the ATP/AMP ratio, and the activities of complexes I and IV. In conclusion, metformin could attenuate the HC diet-induced redox imbalance, inflammation, and mitochondrial dysfunction in blunt snout bream.

This study evaluates using different levels of the white button mushroom powder (WBMP) on some mucosal innate immune parameters (lysozyme, protease, esterase, alkaline phosphatase activities, and total immunoglobulin levels), and the relative expression of some principal immune-relevant genes (lysozyme, TNF-α, and IL-1β) in the zebra danio intestine. Zebrafish specimens (1.75 ± 0.25 g) were divided into experimental units based on the additives to a diet including 5, 10, and 20 g of WBMP per kilogram of food weight, alone or in conjunction with the antibiotic (10 mg/kg BW), and the AGRIMOS (1 g/kg food weight). Following the 11-day experimental duration, the skin mucus and intestine were sampled. To assess the immune gene expression, the real-time PCR detection system was conducted according to the ΔΔCt method using the IQ5 software (Bio-RAD). Results showed that all groups had a significant increase in terms of mucosal lysozyme activity compared to the control group. Examination of total immunoglobulin, protease, esterase, and ALP activity in fish under experimental treatment showed that there was no significant difference between the trial groups and the control groups. The most expression of the lysozyme gene was related to the group that was separately taken the lower concentration (5 g per kg of FW) of WBMP. In conclusion, the amount of 1% mushroom powder in the diet can improve its immune function. Our recommendation is that given the positive effects that mushroom powder added on the diet alone, avoid taking antibiotics for this purpose.

A 90-day feeding trial was conducted to evaluate the effects of replacing dietary fishmeal (FM) with defatted black soldier fly larvae meal (BSFL) on the growth performance and fillet quality of largemouth bass (Micropterus salmoides). The largemouth bass was divided into six groups (BSFL0, BSFL15, BSFL30, BSFL45, BSFL75, and BSFL100) and fed six isonitrogenous(CP 50%, 508 g/kg) and isolipid (CL 9%, 124 g/kg) diets, in which 0, 15%, 30%, 45%, 75%, and 100% of the fishmeal was replaced with BSFL, respectively. The results showed that the final body weight (FBW) and specific growth rate (SGR) of the largemouth bass decreased with increasing BSFL content, and they were significantly lower in BSFL75 than in BSFL0. The weight gain rate (WGR) decreased with increasing BSFL content and the feed conversion ratio (FCR) of largemouth bass increased with increasing BSFL content. The saturated fatty acids (SFAs) and n-3 polyunsaturated fatty acids (PUFA) contents of the largemouth bass fillet significantly decreased, and the n-6 PUFA content of the largemouth bass fillets significantly increased with increasing dietary BSFL. The fillet b* significantly decreased with increasing BSFL content. The biological parameters, fillet proximate nutrient composition, fillet amino acid composition, skin color, and fillet texture of the largemouth bass were not affected by the replacement of BSFL. Transcriptomic analysis revealed that BSFL replacement of FM affects the immune system and metabolic processes of largemouth bass through signaling pathways such as complement and coagulation cascades, the PPAR signaling pathway, cholesterol metabolism, and fat digestion and absorption. In conclusion, a replacement level lower than 45% BSFL was suggested for the overall growth and fillet quality of largemouth bass.

During the spring of 2022, several endangered leuciscid species (Anaecypris hispanica, Squalius aradensis, Anachondrostoma Occidentale, and Iberochondrostoma lusitanicum) were sampled both at the Vasco da Gama aquarium facilities and in some rivers of the Algarve region, Portugal. Sperm samples were extracted by gentle abdominal pressure and sperm motion parameters were assessed for the first time in four species, using a computerized analysis system. The results obtained showed that spermatozoa kinetic patterns were similar for all 4 species, with high motility and velocity values after the sperm activation time and with a marked decrease after 20. On the other hand, sperm longevity was highly variable between species, with short longevities (around 40 s) for A. hispanica and S. aradensis, and longer longevities (100-120 s) for A. occidentale and I. lusitanicum, which could indicate a latitudinal pattern in terms of sperm longevity. At the same time, morphometric analysis was carried out for the four target species, revealing that spermatozoa showed similar sizes and shapes to other external fertilizers belonging to Leuscididae, with small spherical heads, uniflagellate, and without acrosomes. In addition, a short-term gamete storage trail was performed by diluting sperm in 1:9 (sperm:extender) and storing them at 4ºC. Although the results obtained were uneven among the species studied, the dilution and extender used generated motilities above 40% up to day 4 of storage in S. aradensis and I. lusitanicum, and up to days 1-2 in A. hispanica and A. occidentale, respectively. Finally, gamete cryopreservation trials were also carried out on these threatened species. Although cryopreserved samples showed significantly lower motility than fresh samples, some protocols generate acceptable percentages of viability, DNA integrity, and sperm motility in some species such as I. lusitanicum and A. occidentale. The data revealed that the protocol based on 10% DMSO plus 7.5% egg yolk generated the best results.This study is the first to assess the reproductive traits of wild and captive populations of endangered leuciscids endemic from the Iberian Peninsula, describing the spermatozoa kinetics and developing protocols for managing male gametes both in short- and long-term storage. Outcomes will provide new and useful tools to complement the management and conservation of ex situ breeding programs that are being developed for these four endangered species.

Induced development of haploid embryos (H) with only paternal (androgenesis) or maternal (gynogenesis) chromosomes requires irradiation of eggs before fertilization or activation of eggs with irradiated spermatozoa, respectively. To provide doubled haploids (DHs), androgenetic and gynogenetic haploid zygotes need to be subjected to the thermal or high hydrostatic pressure (HHP) shock to suppress the first mitotic cleavage and to double paternal or maternal haploid set of chromosomes. Androgenesis and mitotic gynogenesis (mito-gynogenesis) result in the generation of fully homozygous individuals in a single generation. DHs have been utilized in selective breeding programs, in studies concerning the phenotypic consequences of recessive alleles and to evaluate the impact of sex chromosomes on the early ontogeny. Moreover, the use of DHs for the NGS approach radically improves de novo the assembly of the genomes. However, reduced survival of the doubled haploids limits the wide application of androgenotes and gynogenotes. The high mortality of DHs may be only partly explained by the expression of recessive traits. Observed inter-clutch variation in the survival of DHs developing in eggs originating from different females make it necessary to take a closer look at the quality of the eggs used during induced androgenesis and gynogenesis. Moreover, the developmental competence of eggs that are subjected to irradiation before fertilization in order to deactivate maternal chromosomes when undergoing induced androgenesis and exposed to the physical shock after fertilization that leads to the duplication of the zygotes in both mito-gynogenesis and androgenesis may be also altered as irradiation and sublethal values of temperatures and hydrostatic pressure are considered as harmful for the cell organelles and biomolecules. Here, recently provided results concerning the morphological, biochemical, genomic, and transcriptomic characteristics of fish eggs showing high and low competence for androgenesis and mito-gynogenesis are reviewed.