Mohammad Ali Nematollahi

The effect of Pediococcus acidilactici bacteria used as probiotic supplement on the growth and non-specific immune responses of green terror, Aequidens rivulatus

A 56-day feeding trial was conducted on a species of ornamental fish called green terror (Aequidens rivulatus) (0.388 ± 0.0021 g) to assess the effect of probiotic bacteria, Pediococcus acidilactici on the growth indices and innate immune response. The fish were randomly allocated into 9 oval tanks (120 l) at a density of 60 fish per tank. The experimental diets were comprised of the control (C), C complemented with fish oil (O) and the probiotic and fish oil (PA) and fed ad lib twice a day. The growth indices (specific growth rate (SGR), feed conversion ratio (FCR) and immunological indices of fish fed the diets including lysozyme activity, total immunoglobulin and alternative complement activity were measured. The Fish fed with the diet containing P. acidilactici (PA) displayed significantly (P < 0.05) higher final weight (3.25 ± 0.065 g), weight gain (830.94 ± 9.46%), SGR (3.53 ± 0.02%/day) and lower FCR (1.45 ± 0.011) compared to those of other experimental diets. Total immunoglobulin (10.05 ± 0.12 μg/ml), lysozyme activity (4.08 ± 0.85 μg/ml) and alternative complement activity (2.65 ± 0.12 U/ml) in the serum of PA fed fish showed significant compared to other treatments (P < 0.05). The results showed positive effects of P. acidilactici as a potent probiotic on growth indices and non-specific immune system of green terror.

Transcript levels of five enzymes involved in cortisol synthesis and regulation during the stress response in common carp: Relationship with cortisol

In this study the expression of five genes involved in cortisol synthesis and regulation in the head kidneys of common carp (Cyprinus carpio L.) has been investigated in response to 3h net confinement stress, followed by 22h recovery. Cortisol, glucose, lactate and free fatty acid levels were measured in blood plasma. StAR, P450c17a2, 3betaHSD, P450c21 and 11betaHSD2 transcript sequences were identified based on Cyprinidae homologs and quantified by real-time PCR. Results showed that the plasma cortisol level reached a peak at one hour post-stress (85-fold higher than in control) and quickly returned to normal after 4h recovery. 11betaHSD2 transcripts were for the first time identified in interrenals. Changes in cortisol levels during and after confinement were correlated in a time-delayed relationship with increase and decrease in mRNA levels of 11betaHSD2, respectively. These results suggest that cortisol may be involved in the control or activation of 11betaHSD2. StAR and P450c21 mRNA levels did not change during net confinement stress and recovery, but P450c17a2 levels were significantly increased 4 and 22h after recovery. Since plasma cortisol levels increased by 68-fold within 5min net confinement stress, it seems that transcriptional activation of this enzyme is not directly involved in acute cortisol production.

IGF-I and IGF-II effects on local IGF system and signaling pathways in gilthead sea bream (Sparus aurata) cultured myocytes

The insulin-like growth factors (IGFs) have a fundamental role in a vast range of functions acting through a tyrosine-kinase receptor (IGF-IR). IGFs in muscle can affect the expression of components of the local IGF system, myogenic regulatory factors (MRFs), proliferating (proliferating cell nuclear antigen, PCNA) or differentiating molecules (myosin heavy chain, MHC) and, lead to the activation of different signaling pathways. The response of all these genes to IGFs incubation at two different times in day 4 cultured myocytes of gilthead sea bream was analyzed. Both IGFs increased the expression of IGF-I and IGFBP-5, but showed different effects on the receptors, with IGF-I suppressing the expression of both isoforms (IGF-IRa and IGF-IRb) and IGF-II up-regulating only IGF-IRb. Moreover, the protein levels of PCNA and target of rapamycin (TOR) increased after IGF-II incubation, although a decline in Myf5 and a rise in MHC gene expression was caused by IGF-I. Taken together, these results provide evidence for the importance of IGFs on controlling muscle development and growth in gilthead sea bream and suggest that each IGF may be preferentially acting through a specific IGF-IR. Moreover, the data support the hypothesis that IGF-II has a more important role during proliferation, whereas IGF-I seems to be relevant for the differentiation phase of myogenesis.

The transcription of l-gulono-gamma-lactone oxidase, a key enzyme for biosynthesis of ascorbate, during development of Persian sturgeon Acipenser persicus

l-Gulono-gamma-lactone oxidase (GULO) is a key enzyme for the biosynthesis of ascorbate, which is essential for several cellular functions. In the present study, mRNA expression of GULO gene was evaluated during the early development of Persian sturgeon. First, because there are no comparative studies that have established suitable quantitative real-time PCR reference genes in sturgeons for any physiological conditions, we evaluated six candidate reference genes (ACTB, RPL13, UBQ, RPL6, GAPDH and EF1A) during the early development of Persian sturgeon. The most stable mRNA expression was obtained with RPL6 and ACTB, whereas the least stable was RPL13. After normalization using RPL6, ACTB and RPL6/ACTB combination, the mRNA expression of GULO was highest at the embryonic stage (2days before hatching; P<0.05) and started to decline from hatching of larvae to the rest of the developmental time-points. This suggests that the vitamin C requirements are highest during early life stages, and it is likely that the changes in GULO mRNA expression are associated with changes in GULO enzyme activity.

High levels of corticosterone, and gene expression of star, cyp17a2, hsd3b, cyp21, hsd11b2 during acute stress in common carp with interrenal hyperplasia

We investigated the acute stress response in a common carp strain (E5) with interrenal hyperplasia due to 17α-hydroxylase deficiency, and in an isogenic standard (STD) carp strain. Cortisol, corticosterone and the head kidney-somatic index were measured during and after a 3 h net confinement stress. Star, cyp17a2, hsd3b, cyp21, hsd11b2 mRNA levels were measured in head kidneys using real-time qPCR. The results show very high corticosterone levels and enlargement of the head kidney in E5 fish. This is the first report in a teleost fish showing a significant increase of corticosterone levels in response to stress due to interrenal hyperplasia. The high levels of corticosterone in E5 suggest that corticosterone is not converted to aldosterone in common carp. star and hsd3b mRNA levels were significantly higher in E5 compared to STD fish, while cyp17a2 levels were significantly lower in E5. In contrast to E5, star levels did not change during stress and recovery in STD, suggesting that the enzyme is regulated in a different manner in E5 and STD fish. In E5, the levels of cyp17a2 dropped below control values after 20 min stress. These findings strongly suggest that cyp17a2 is impaired at (post)-transcriptional level. As a consequence the accumulated precursor (pregnenolone) is not converted to cortisol, but to corticosterone. In contrast to STD, significant levels of cortisol could not be detected in E5. Finally, hsd11b2 mRNA levels were significantly lower in E5 compared to STD, and did not change during stress and recovery. These results support the idea that hsd11b2 is involved in the conversion of physiologically active cortisol to inactive cortisone, as reported earlier for STD carp. In conclusion our results show high levels of corticosterone in E5 and differences in star and mRNA levels of steroidogenic genes between E5 and STD carp during net confinement stress.

Lysine and Leucine Deficiencies Affect Myocytes Development and IGF Signaling in Gilthead Sea Bream (Sparus aurata)

Optimizing aquaculture production requires better knowledge of growth regulation and improvement in diet formulation. A great effort has been made to replace fish meal for plant protein sources in aquafeeds, making necessary the supplementation of such diets with crystalline amino acids (AA) to cover the nutritional requirements of each species. Lysine and Leucine are limiting essential AA in fish, and it has been demonstrated that supplementation with them improves growth in different species. However, the specific effects of AA deficiencies in myogenesis are completely unknown and have only been studied at the level of hepatic metabolism. It is well-known that the TOR pathway integrates the nutritional and hormonal signals to regulate protein synthesis and cell proliferation, to finally control muscle growth, a process also coordinated by the expression of myogenic regulatory factors (MRFs). This study aimed to provide new information on the impact of Lysine and Leucine deficiencies in gilthead sea bream cultured myocytes examining their development and the response of insulin-like growth factors (IGFs), MRFs, as well as key molecules involved in muscle growth regulation like TOR. Leucine deficiency did not cause significant differences in most of the molecules analyzed, whereas Lysine deficiency appeared crucial in IGFs regulation, decreasing significantly IGF-I, IGF-II and IGF-IRb mRNA levels. This treatment also down-regulated the gene expression of different MRFs, including Myf5, Myogenin and MyoD2. These changes were also corroborated by a significant decrease in proliferation and differentiation markers in the Lysine-deficient treatment. Moreover, both Lysine and Leucine limitation induced a significant down-regulation in FOXO3 gene expression, which deserves further investigation. We believe that these results will be relevant for the production of a species as appreciated for human consumption as it is gilthead sea bream and demonstrates the importance of an adequate level of Lysine in fishmeal diet formulation for optimum growth.

Quantity discrimination in parental fish: female convict cichlid discriminate fry shoals of different sizes

Numerical abilities have been found to be adaptive in different contexts, including mating, foraging, fighting assessment and antipredator strategies. In species with parental care, another potential advantage is the possibility to adjust parental behavior in relation to the numerosity of the progeny. The finding that many fish vary their parental investment in relation to brood size advocates the existence of a mechanism for appraising offspring number, an aspect that has never been directly investigated. Here we tested the ability of parental female convict cichlid (Amatitlania nigrofasciata) to discriminate between two fry groups differing in number by measuring time spent attempting to recover groups of fry experimentally displaced from the next. Females spent more time trying to recover the fry from larger groups when tested with contrasts 6 versus 12 (1:2) and 6 versus 9 fry (2:3); however, they showed no preference in the 6 versus 8 (3:4) contrast, suggesting that this task exceeds their discrimination capacity.

The toxicological effect of Ruta graveolens extract in Siamese fighting fish: a behavioral and histopathological approach

The effects of pharmacological waste on aquatic ecosystems are increasingly highlighted in ecotoxicology research. Many of these products are designed for human physiology but owing to the conservative nature of vertebrate evolution they also tend to have effects on aquatic organisms and fishes in particular when they find their way into aquatic systems via wastewater effluent. One area of research has focused on reproductive control and the associated hormone treatments. Many of these hormones affect the reproductive physiology of fishes and may cause feminization of male reproductive traits. Alternative medicines have also been widely used particularly in traditional cultures but few of these alternative treatments have been assessed with respect to their potential impact on aquatic ecosystems. Rue (Rutagraveolens) has been used as a male contraceptive in traditional medicines but its effects on fish behavior and reproductive anatomy have yet to be established. Here we show that treating Siamese fighting fish, Bettasplendens, with extract of rue has a significant effect on key aggressive/reproductive behaviors and the propensity to explore novel objects (boldness). In all cases the respective behaviors were reduced relative to controls and sham injected fish. Histological analysis of the testes revealed that rue exposure reduced the number of spermatozoa but increased the number of spermatocytes relative to controls.

Characterization data of gilthead sea bream (Sparus aurata) IGF-I receptors (IGF-IRa/Rb)

In this data article we describe the coding sequence of two IGF-IR paralogues (IGF-IRa and IGF-IRb) obtained from gilthead sea bream embryos. The putative protein architecture (domains and other important motifs) was determined and, amino acid sequences alignment and phylogenetic analysis of both receptors together with IGF-IR orthologues from different vertebrates was performed. Additionally, a semi-quantitative conventional PCR was done to analyze the mRNA expression of both receptors in different tissues of gilthead sea bream. These data will assist in further physiological studies in this species. In this sense, the expression of both receptors during ontogeny in muscle as well as the differential effects of IGF-I and IGF-II on their regulation during in vitro myogenesis has been recently studied (doi: 10.1016/j.ygcen.2015.11.011; [1]).

Effects of prebiotic mannan oligosaccharide on the growth, survival, and anxiety-like behaviors of zebrafish (Danio rerio)

ABSTRACT A study was designed to investigate the possible positive prebiotic effects of mannan oligosaccharides (MOS) on growth and reduction of anxiety behaviors in zebrafish (Danio rerio). The experimental diets were supplemented with 0, 2, 4, 6, or 8 g MOS kg−1 and were fed to juvenile fish (0.47 ± 0.017 g) in triplicates for six weeks. Significant differences were found in the final body weight, body weight intake, percent body weight intake, and specific growth rate among treatments. The regression analyses showed that the optimum MOS level was 4–4.1 g MOS kg−1 diet. Fish fed 4 g MOS kg−1 feed showed higher survival than the other treatments. Also, using the novel tank test, fish fed with 4 g MOS kg−1 diet showed lower anxiety by swimming in the upper portion of the tank. The other behavior parameters remained unaffected by the prebiotic. The results indicated the beneficial effect of the prebiotic MOS on growth and partly on welfare of zebrafish juveniles.