The effects of diazinon on the cell types and gene expression of the olfactory epithelium and whole-body hormone concentrations in the Persian sturgeon (Acipenser persicus).

The olfactory function and imprinting of odorant information of the native stream play a critical role during the homing migration in fish. Pesticides may impair olfactory imprinting by altering olfaction and hormone functions. The present study aimed to determine how diazinon impacts olfactory epithelium morphology and cell composition, as well as hormone concentrations in Persian sturgeon (Acipenser persicus) during their lifetime in freshwater and, also during diazinon-free saltwater acclimation. Fingerlings were exposed to 0, 150, 300, and 450 µg·L-1 of diazinon in freshwater for 7 days and then were transferred to diazinon-free saltwater by gradually increasing salinity up to 12 ppt. After diazinon exposure, the number of olfactory receptor cells (ORCs) and goblet cells (GCs) decreased and increased, respectively, and the expression of G-protein αolf (GPαolf) and calmodulin-dependent kinase II delta (CAMKIId) was down-regulated and up-regulated, respectively. Transferring the fish to diazinon-free saltwater (8 and 12 ppt) raised the number of ORCs, supporting cells (SCs), GCs, and GPαolf expression, and down-regulated CAMKIId without any significant differences among treatments. Exposure to diazinon increased whole-body cortisol at the high concentration, while decreased whole-body thyroxin (T4) and triiodothyronine (T3) in a dose-dependent manner. Although whole-body T4 and T3 increased in all the treatments after saltwater acclimation (8 and 12 ppt), the level of these hormones was lower in fish that had been exposed to diazinon than in the control. These results showed that diazinon can disrupt olfactory epithelium morphology and cell composition as well as hormone concentrations, which in turn may affect the olfactory imprinting in Persian sturgeon fingerlings.

Droplet vitrification versus straw cryopreservation for spermatozoa banking in Persian sturgeon (Acipenser persicus) from metabolite point of view.

Persian sturgeon (Acipenser persicus), a commercially valuable and critically endangered fish species has been suffering considerable declines in populations in the nature due to over-fishing, habitat destruction and marine pollution during past decades. Since there were no achievements in artificial reproduction programs, genetic resource banking such as gametes and embryo cryopreservation can be a good strategy however, reported resulting gamete qualities were considerably low. In the present study, the metabolome content of Persian sturgeon spermatozoa was investigated during common straw cryopreservation and novel droplet vitrification by the use of 1H NMR (Nuclear Magnetic Resonance) spectroscopy. Univariate (ANOVA) and multivariate (PCA) analysis showed significant differences in the metabolic profiles between cryopreserved and fresh spermatozoa samples. Adenine, creatine, creatine phosphate, glucose, guanidoacetate, lactate, N, N-dimethylglycine, and glycine levels showed no significant differences between these two cryopreservation techniques suggesting these metabolites and their corresponding enzymes and chemical pathways are so vulnerable to the temperature changes and even higher cooling rate in droplet vitrification could not conserve them. However, significant differences were found in acetate, creatinine, betaine, ß-alanine and trimethylamine N-oxide suggesting better efficiency of droplet vitrification in protection of some metabolites associated to spermatozoa energetics, redox balance and hypoxia compensation compared to straw cryopreservation.

The effect of chlorpyrifos on salinity acclimation of juvenile rainbow trout (Oncorhynchus mykiss).

As a part of their unique life cycle, most salmonids undergo a transition from fresh water to salt water requiring various adjustments in metabolism, osmoregulation and ion regulation. Exposure to pesticides may affect the acclimation of juvenile salmonids to salt water during downstream migration to estuaries. Using the Caspian Sea as a model waterbody, the present study aimed to determine how the toxicity of the organophosphate pesticide chlorpyrifos (CPF) impacts saline acclimation of rainbow trout (Oncorhynchus mykiss). We pre-exposed 4-month-old fish to nominal concentrations of 0, 20, 40, 80, 160 µg/L of CPF for seven days, and then gradually to salinity (12 ppt) for another seven days. Mortality, levels of cortisol, T3 and T4 in serum, and expression of genes involved in gill ion transport (Na+/K+ATPase α1a and α1b) and liver xenobiotic detoxification (Glutathione-S-Transferase pi, GST) were measured at day fourteen. Cortisol concentrations in serum were not changed by CPF exposure in freshwater, but serum T3 increased up to three fold relative to controls in freshwater. Following salinity acclimation, T3 and T4 concentrations in the serum were both increased up to 2.5 and 8.8 fold in animals treated with CPF followed by saltwater. Na+/K + ATPase α1a and α1b mRNA in gill were unchanged by CPF treatment in freshwater but trended higher in CPF-treated animals after salinity acclimation. Hepatic mRNA of GST was significantly increased following exposure to CPF but was unchanged after saltwater exposure. Although saltwater treatment reduced the acute lethality of CPF, changes in T3/T4 suggest sublethal impacts may occur in CPF-treated fish after they acclimate to Caspian seawater.

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]).

Fluoxetine inhibits aggressive behaviour during parental care in male fighting fish (Betta splendens, Regan).

The increasing presence of aquatic contaminants, such as the pharmaceutical fluoxetine, has raised concerns over potentially disrupting effects on several aspects of fish reproduction. However, the effects of fluoxetine on reproductive and paternal behavior in fish remain understudied, particularly at environmentally relevant concentrations. In the current study, we therefore tested the hypothesis that waterborne fluoxetine at an environmentally relevant concentration (540 ng/l), disrupts specific reproductive and paternal behaviors in male Siamese fighting fish at distinct reproductive phases. A pre-post test design was adopted to investigate specific behavioral responses at the individual fish level in response to male conspecific intruders at two different distances from the nest across four distinct reproductive phases (before bubblenest construction, following bubblenest construction, after spawning and after hatching of the larvae). In the control specimens, the measured behaviours were not different between the spawning times and among the interactions in either distance to nest at the different reproduction phases. Our results indicate that fluoxetine specifically disrupts characteristic paternal territorial aggression behaviour only after spawning and hatching of the larvae, while male behaviour in previous reproductive phases is unaffected by fluoxetine exposure. Results of comparison between males at 1st spawning and specimens exposed to fluoxetine at 2nd spawning showed that the first reaction of the nest-holding males to the intruders, duration of fin spreading, number of bites, and 90° turn, and the frequency of sweeps were different between the spawning times after spawning or hatching of embryos. However, interaction of spawning time and reproduction phase was significant on biting behaviour. These results demonstrate that fluoxetine exposure at environmental concentrations negatively affects territorial defense behaviour in fighting fish during parental care after larval hatching, which may have possible implications on reproductive success and population dynamics.

Impact of a short-term diazinon exposure on the osmoregulation potentiality of Caspian roach (Rutilus rutilus) fingerlings.

The stocks of Caspian roach (Rutilus rutilus), an economically important species in the Caspian Sea, are depleting. Each year millions of artificially produced fingerlings of this species are restocked in the mouth of rivers of the Southern Caspian Sea (e.g. Qare Soo River), where they are exposed to pesticides originating from regional rice and orchard fields. This early exposure to pesticides could affect the hypo-osmoregulatory ability of juvenile fish. Thus, in this study, Caspian roach fingerlings were exposed to environmentally-relevant concentrations of the organophosphate insecticide diazinon for 96 h in fresh water and then transferred to diazinon-free brackish water (BW) for another 96 h. We report that cortisol and glucose levels were significantly increased in all diazinon treatments at all sampling time points in comparison to the control group. Moreover, the thyroid hormone levels of TSH, T4, and T3 significantly decreased in diazinon-exposed fish even after the transfer to BW. The electrolytes were differentially affected during the exposure to diazinon and after the transfer to BW. The number of chloride cells in the gill tissue was significantly increased during diazinon exposure at the higher concentrations and decreased to control levels after transfer to BW. Finally, gill and kidney tissues showed many histopathological changes in diazinon-exposed fish even after 240 h in BW. These results suggest that the release of Caspian roach fingerlings into the diazinon-contaminated Caspian Sea regions may alter their physiology and jeopardize their survival, which could lead to a failure in rebuilding the Caspian roach stocks in the Caspian Sea.

The study of some haematological and serum biochemical parameters of juvenile beluga (Huso huso) fed oligofructose.

A study was conducted to investigate the effects of dietary oligofructose (1, 2 and 3%) on the blood profiles of beluga (Huso huso) juveniles (18.77 ± 0.76 g) compared to fish fed an un-supplemented diet. After 7 weeks of feeding on the experimental diets, haematological parameters, metabolic products (cholesterol, glucose and total protein) and serum enzymes (lactate dehydrogenase, alkaline phosphatase, alanine aminotransferase and aspartate aminotransferase) were measured. Compared to the control group (0% oligofructose), dietary oligofructose had no effect on red blood cell counts (RBC), mean corpuscular volume (MCV), mean cellular haemoglobin (MCH) or mean cell haemoglobin concentration (MCHC) (P > 0.05). However, haemoglobin (Hb) concentration, leucocyte (WBC) levels and the proportion of lymphocytes were significantly higher (P > 0.05) in the 2% oligofructose fed fish than in the 3% oligofructose fed fish. Additionally, haematocrit (Hct) values (P = 0.049) and the proportion of lymphocytes (P ≤ 0.01) were significantly higher in the 2% oligofructose group than in the control group. Although serum glucose and total protein remained unaffected, serum cholesterol was significantly lower in the 2% oligofructose group than in the control and 3% oligofructose group (P < 0.05). The results of the present study showed that oligofructose had no significant effects on serum lactate dehydrogenase, alkaline phosphatase, alanine aminotransferase and aspartate aminotransferase. These results indicate that fish blood profiles could be affected by prebiotics, which should be taken into account in future studies.

Effects of triploidy on the Caspian salmon Salmo trutta caspius haematology.

The aim of this study was a comparison of key haematological features of diploid (2n) and triploid (3n) Caspian salmon (Salmo trutta caspius). Morphometric indices of erythrocytes were determined on blood smears by light microscopy. Triploidy significantly (P < 0.001) increased all morphometric indices measured in the erythrocytes including cell size, cell surface area, and cell volume. The increase in cell size was larger for the major (27%) axis than for the minor (22%) axis, thus making erythrocytes of 3n Caspian salmon more ellipsoidal. The estimated increase in erythrocyte nuclear volume (87%) was bigger than the theoretical expected 50% increase. Haematological indices were measured manually by hemocytometry. Triploids had lower numbers of red blood cells (RBC: 1,120,000 cells/mL in 2n vs. 700,000 cells/mL in 3n; P < 0.001) but they were larger in size (mean erythrocytic volume [MEV]: 363.1 nm3 in 2n vs. 483.3 nm3 in 3n; P < 0.001). The decrease in RBC number was not compensated by the increase in MEV and, thus, triploidy affected the haematocrit (Hct: 38.8% in 2n vs. 33.06% in 3n; P < 0.05). Total blood hemoglobin concentration was lower in triploid fish (Hb: 9.9 g/dL in 2n vs. 8.9 g/dL in 3n; P < 0.05). In contrast, mean erythrocytic hemoglobin (MEH: 95 mug in 2n vs. 133.2 mug in 3n; P < 0.001) was higher for 3n Caspian salmon as a result of their larger erythrocytes, although MEH concentration (MEHC: 0.26 g/dL in 2n vs. 0.27 g/dL in 3n) did not significantly differ (P > 0.05). White blood cell (WBC) counts (lymphocytes and neutrophiles) were measured and WBC/RBC ratios were calculated. There were no significant differences in WBC (15,710 cells/mL in 2n vs. 12,683 cells/mL in 3n; P > 0.05), lymphocytes, and neutrophils as %WBC as well as WBC/RBC ratios between two ploidy levels (P > 0.05). Triploid Caspian salmon showed higher erythrocyte abnormalities such as 'twisted', 'tailed', and 'anucleated' cells as well as high portions of immature RBC in blood smears in comparison with diploids (P < 0.001).

Spermatozoa motility in the Persian sturgeon, Acipenser persicus: effects of pH, dilution rate, ions and osmolality.

Sperm motility is a prerequisite factor determining semen quality and fertilizing capacity. The effects of environmental factors including pH, cations and osmolality as well as the role of dilution rate on sperm motility parameters in Acipenser persicus were studied. The best pH and dilution rate for activation of spermatozoa were pH 8.0 and dilution ratio 1:50. Ionic factors can stimulate the initiation of sperm activation. The maximum percentage of motile sperm and total duration of sperm motility were observed in solutions containing 25 mM NaCl, 0.2 mM KCl, 3 mM CaSO4, 10 mM MgSO4 and sucrose with an osmolality of 50 mosmol kg(-1). The present study provides us with some basic knowledge about sturgeon spermatozoa biosensitivity to ionic and osmolality effects. A sensitivity of A. persicus sperm was observed after induction of activation of sperm motility in solution containing cations or sucrose with high osmolality. Concentrations more than 50 mM Na+, more than 1 mM K+, more than 3 mM Ca2+ and more than 10 mM Mg2+ had negative effects on sperm motility. Also, osmolality more than 100 mosmol kg(-1) had an inhibitory effect. It is clear that ions and osmolality stimulate the motility of spermatozoa by changes in the properties of the plasma membrane including its potential and its ionic conductance. The inhibitory role of high osmolality of the swimming medium (more than 100 mosmol kg(-1)) and insufficient osmolality of the seminal plasma to inhibit semen motility suggested that osmolality is not the principal factor preventing sperm motility in seminal fluid but that K+ is a major inhibitory factor of sperm motility in seminal plasma.