Effect of Zataria multiflora essential oil and potassium sorbate on inoculated Listeria monocytogenes, microbial and chemical quality of raw trout fillet during refrigerator storage.

Human listeriosis is predominantly associated with contaminated food consumption, including seafood, shrimp, and RTE foods. Listeria monocytogenes is a foodborne pathogen that is mainly found in freshwater, seawater, and fish mucus. Seafood contamination can occur during food processing. L.monocytogenes levels of below 100 cfu/g can be found in seafood samples. The present study was conducted to investigates the effect of Zataria multiflora essential oil (ZEO) and potassium sorbate (PS) on microbial and chemical changes in raw rainbow trout at 4°C to extent shelf life and improve food safety. First, the chemical compositions of ZEO were identified. Then, different percentage of ZEO (1.5, 0.8, and 0.5%) and PS (2%) were inoculated in raw fish fillets and analyzed for TVC, TBA, TVB-N, pH, sensory attributes, Pseudomonas aeruginosa, and inoculated L. monocytogenes (1 × 105 cfu/g) survival at 4°C for 12 days. The best sensory evaluation score was observed for the samples treated with 0.8% and 1.5% ZEO. Overall, this study results indicated that the treatment of rainbow trout fillet with 1.5% ZEO is the best method for controlling the growth of L. monocytogenes at refrigerator temperature without any undesirable sensory effects.

Effects of nanosilver on hematologic, histologic and molecular parameters of rainbow trout (Oncorhynchus mykiss).

Efficient antibacterial and antifungal properties of silver nanoparticles (AgNPs) sparked its commercial application in several industrial and household products. Drastic increase of AgNPs production raised concerns over aquatic organisms' exposure. The toxic dose, mechanism of toxicity, physiological damages, gene expression alteration, hematological and blood parameter distortion by AgNP needs to be investigated to explore inevitable risk in aquatic animals. In this study, rainbow trout (Oncorhynchus mykiss) (122.4 ± 1.4 g, 23.8 ± 0.7 cm) were exposed to colloidal AgNPs (28.3 ± 12.6 um) to determine the lethal concentration (LC50)(8.9 mg/l). Sub-lethal concentrations (10 %LC50, 25 %LC50, plus LC50 value) impact on hematologic, histological and molecular responses were evaluated. Results showed sever damage to blood cells morphology, and hematologic parameters change including RBC, WBC, Hct and Hb in all AgNP-treated groups. Histological damage in gill and liver of exposed fish were observed. Significant up-regulating of HSP70 and P53 genes were detected in response to AgNPs, whereas, it was found that in comparison to HSP70 gene, P53 induction occurred in lower AgNPs concentrations and lower exposure time. These results indicate adversely effects of AgNPs exposure to aquatic environments.

Transgenerational disrupting impacts of atrazine in zebrafish: Beneficial effects of dietary spirulina.

In a range of fish species, offspring sustainability is much dependent to their mother's investment into the egg yolk. A healthy environment helps broodfish to produce normal quality offspring. However, deviation from optimal conditions can disturb body functions that effect the next generation. Here, zebrafish (Danio rerio) was employed to investigate the transgenerational impacts of an immunotoxic and endocrine disruptor, atrazine (AZ). In addition, the possible ameliorated effects of a nutraceutical, Arthrospira platensis (spirulina- SP), was considered. Adult females were either exposed to 0 (Cn), 5 (AZ5), and 50 (AZ50) µg/L AZ or fed SP-supplemented diet (10 g/kg; SP). In combination treatments, fish were also exposed to AZ and fed SP (SP-AZ5 and SP-AZ50). Embryos were obtained after 28 d of exposure. Exposure to AZ50 caused females to produce eggs with significantly lower fertilization and hatching. No changes were observed in the concentrations of thyroid hormones. AZ significantly increased cortisol response and reduced levels of immunoglobulin, lysozyme and complement activities in females and their offspring. SP-AZ5 and SP-AZ50 females, however, resisted to the toxic effects of AZ, produced embryos with lower cortisol content and higher immunity competence. Bactericidal activity of the embryos also showed the transgenerational antimicrobial effects of SP along with the AZ immunotoxicity. Overall, these results indicate that AZ could have long lasting toxic effects on fish, and that dietary SP-supplementation could ameliorate AZ induced transgenerational toxic effects.

A bioassay on tissue cholinesterase activity of Rutilus kutum (Kamensky, 1901) exposed to some common pesticides in Iran.

The toxicological effects of three commonly used pesticides in Iran on the fries of Caspian kutum (Rutilus kutum, Kamensky, 1901) were investigated through determining 50.00% lethal concentration (LC50) 96hr and cholinesterase (ChE) inhibition. The LC50 96-hr of carbaryl, glyphosate, and malathion were equal to 11.69, 6.64, and 0.97 mg L-1, respectively, that were considered as harmful (10.00 - 100 mg L-1), toxic (1.00 - 10.00 mg L-1) and very toxic (< 1.00 mg L-1) compounds for this species. The exposure of fries to sub-lethal concentrations of the pesticides over 15 days indicated that the average of ChE activity in the head and trunk were 1086.89 ± 124.34 and 627.36 ± 99.60 mU min-1 per mg protein, respectively, with a significant difference relative to each other. There was a significant difference between fry exposed to all three pesticides and the control group in cholinesterase inhibition. The fries exposed to carbaryl (890.12 ± 28.08 mU min-1 per mg protein) and glyphosate (891.77 ± 31.61 mU min-1 per mg protein) showed lower ChE inhibition than those exposed to malathion (790.00 ± 58.14 mU min-1 per mg protein).

DNA damage, acetylcholinesterase activity, and hematological responses in rainbow trout exposed to the organophosphate malathion.

Effects of sub-lethal concentrations (0 (control), 0.009, 0.014, and 0.023 ppm) of the organophosphate insecticide "malathion" to rainbow trout (Oncorhynchus mykiss) after the determination of LC50-96 h value (0.093 ppm) were evaluated. Changes in biomarkers of neurotoxicity (acetylcholinesterase (AChE) activity), genotoxicity (DNA damage), and hematological parameters (red (RBC) and white (WBC) blood cell count, hemoglobin (Hb), hematocrit (Hct), mean cell hemoglobin (MCH), mean cell volume (MCV), and mean cell hemoglobin concentration (MCHC)) were assessed for a 15-day exposure. A significant time- and dose-dependent reduction in AChE activities of gill, muscle, brain, and liver tissues was found. However, the AChE activity was less affected by malathion concentration than by exposure time. DNA damage of erythrocytes at different malathion concentrations increased by increasing the experimental time up to the fourth day. A decrease in the count of WBC, RBC, and Hct and an increase in the number of MCH and MCV were observed by increasing malathion exposure dose and time (p < 0.05). An increase in the malathion concentration and exposure time significantly resulted in a decrease in Hb and an increase in MCHC. A significant improvement in AChE activity; DNA damage; and RBC, Hb, Hct, MCV, and MCH indices was detected during a 30-day recovery period, but the WBC count changed insignificantly. The recovery pattern based on 100% water exchange with clean water could be a successful strategy to improve the biomarker responses of rainbow trout habituating in contaminated aquatic environments.

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.

1H NMR metabolic profiling of the cryopreserved spermatozoa of the wild endangered Persian sturgeon (Acipenser persicus) with the use of beta-cyclodextrin as an external cryoprotectant.

Persian sturgeon (Acipenser persicus) is an endangered species and genetic resource banking such as gametes and embryo preservation could be one of the most pursued conservation approaches. In this study, deleterious effects of the traditional cryopreservation technique and the effect of different doses of 2-hydroxypropyl-beta-cyclodextrin (HßCD) on thawed spermatozoa quality (motility duration and percentage) of Persian sturgeon (Acipenser persicus) were investigated from metabolic aspects of view. For cryopreserving, semen was diluted with Tris-HCl (100 mM) extenders containing 0, 5, 10, and 15 mM of HßCD in a ratio of 1:1 (semen/extenders). Semen-extenders were filled into 0.5-mL straws and were frozen with the vapor of liquid nitrogen, and then immersed into liquid nitrogen. Cryopreserved spermatozoa were thawed in water baths in 15 s. Two treatments with the highest and the lowest motility percentages (0 and 10 mM of HßCD) were chosen to reveal the extremes of the metabolites change range and were objected to 1H NMR spectroscopy. Univariate (ANOVA) and multivariate (PCA) analysis of the obtained metabolic profiles showed significant changes (P < 0.05) in metabolites. The use of 10 mM of HßCD was completely successful in the preservation of creatinine, glucose, guanidoacetate, O-phosphocholine, and N, N-dimethylglycine and probably their corresponding biochemical pathways, but it failed to preserve lactate, carnitine, betain, ß-alanin, and trimethylamine N-oxide. It was also partially successful in preserving acetate, creatine, creatine phosphate, and glycine, all suggesting how HßCD can be effective as a cryoprotectant.

Efficient osmolyte-based procedure to increase expression level and solubility of infectious hematopoietic necrosis virus (IHNV) nucleoprotein in E. coli.

The nucleoprotein of infectious hematopoietic necrosis virus (IHNV) is considered as the main target antigen for detection of IHNV infection in salmonid fish. This study aimed at improving the expression and solubility of IHNV nucleoprotein (IHNV-NP) in E. coli expression system. The effects of several expression strategies including host strain type, protein expression temperature, heat-shock treatment prior to protein induction, and additives in the growth medium and in the cell lysis buffer were examined. Results showed that bacterial strain type had a great impact on protein expression level, whereas it was not effective in preventing protein aggregation. Production of soluble IHNV-NP was proportionally increased with decreased incubation temperature. Heat-shock treatment prior to protein induction did not change the percent of solubility. For cells grown at low temperature, the presence of additives in the lysis buffer enhanced the solubility of IHNV-NP up to 24%. The highest yield of soluble protein was obtained via incorporation of osmolytes in the growth medium of cells exposed to a mild salt stress, in the following order: sucrose > sorbitol > glycerol > glycine. Soluble protein obtained by the optimized condition was efficiently purified in high yield and successfully detected by two monoclonal antibodies in a sandwich ELISA. Taken together, a combination of proper host strain, low-temperature expression, and timely application of osmolytes in the growth medium provided sufficient quantities of soluble recombinant IHNV-NP that has the potential to be used for diagnostic purposes.

In situ assessment of Karaj River genotoxic impact with the alkaline comet assay and micronucleus test, on feral brown trout (Salmo trutta fario).

The in situ evaluation of the genotoxic impact of the Karaj River was performed using the comet and micronucleus (MN) assays in erythrocytes, liver, gill and kidney of indigenous brown trout, Salmo trutta fario from three different stations, including Varangerud, Asara and Purkan. The results showed that DNA damage significantly increased in sampled fish erythrocytes, liver and gill from low levels in the upstream river (Varangerud) via intermediate levels in downstream (Purkan) to high levels in the middle of the river (Asara), correlating with the river increasing pollution gradient. Gill was the most sensitive tissue followed by blood and liver. Kidney did not respond to the genotoxic gradient of the river. MN test (as a complementary assay) of liver cells of fish was a sensitive biomarker of genotoxic exposure. MN test in blood, gill and kidney did not reflect the genotoxic condition of the river.

Silver nanoparticles cause osmoregulatory impairment and oxidative stress in Caspian kutum (Rutilus kutum, Kamensky 1901).

Silver nanoparticles (AgNPs) are increasingly used in several industrial and household products because of their antibacterial and antifungal properties. Hence, there is an inevitable risk that these chemicals may end up in aquatic biotopes and have adverse effects on the fauna. In order to assess potential health effects on aquatic organisms, this study evaluated the effects of waterborne AgNP exposure for 7 days on a set of critical stress parameters in juvenile Caspian kutum (Rutilus kutum), an economically important fish in the Caspian Sea. The applied level 11 µg/l of AgNP is high compared to reported water concentrations and corresponds to 40% of the 96 h LC50 value, initially determined to be 28 µg/l. Gill heat shock protein 70 (hsp70) mRNA expression, Na+/K+-ATPase activity and enzymatic activities of liver superoxide dismutase (SOD), glutathione peroxidase (Gpx), lactate dehyrogenase (LDH) and alkaline phosphatase (ALP), and whole-body cortisol and thyroid hormones (T3 and T4) were measured as endpoints. Gill hsp70 mRNA expression increased and gill Na+/K+-ATPase activity decreased in AgNP-exposed fish compared to controls. The specific activities of all liver enzymes decreased significantly compared to controls. Whole-body cortisol and thyroid hormones decreased compared to controls. In conclusion, the study demonstrates that AgNPs cause oxidative stress and gill osmoregulatory disruption in Caspian kutum juveniles.

NMR-based metabolomic study on the toxicological effects of pesticide, diazinon on adaptation to sea water by endangered Persian sturgeon, Acipenser persicus fingerlings.

NMR-based metabolomics was applied to explore metabolic impacts of diazinon on sea water adaptation of Persian sturgeon fingerlings, Acipenser persicus. Fingerlings were exposed to sub-lethal concentrations of diazinon in freshwater (FW) for 96 h (short-term trial) and 12 days (long-term trial) and then exposed in brackish water (BW) (12 mg L-1 salinity) for 24 h. After 96 h and 12 days exposure in FW, identified metabolites (amino acids, osmolytes, energy metabolites) showed different change-patterns compared to control group (P < 0.05) as follow: (A) short-term trial: higher plasma levels of glucose, lactate (in all diazinon-exposed fish), acetate and acetoacetate (in 0.9 mg L-1diazinon treatment); lower levels of creatine (in all diazinon-exposed fish), trimethylamine-N-oxide, choline, taurine, betaine, N,N-dimethylglycine and almost all amino acids in fish exposed to high concentrations of diazinon (0.54 and 0.9 mg L-1 diazinon). (B) Long-term trial: higher plasma levels of lipid oxidation metabolites and almost all amino acids in fish exposed to 0.54 and 0.9 mg L-1 diazinon; lower levels of creatine, trimethylamine-N-oxide, N,N-dimethylglycine, betaine, choline (in all diazinon-exposed fish), glucose (in 0.54 and 0.9 mg L-1diazinon treatments) and taurine (in 0.9 mg L-1 diazinon treatment). When fish were exposed in BW for 24 h, the plasma levels of osmolytes decreased significantly in almost all experimental groups of short-term and long-term trial (P < 0.05). In short-term trial, the plasma levels of glucose in all groups and lactate in 0.18 and 0.54 mg L-1 diazinon treatments increased after salinity challenge (P < 0.05). However, a significant decrease was observed in lactate levels in 0.9 mg L-1 diazinon treatment (P < 0.05). Also, the plasma levels of amino acids decreased mostly in fish of control group than exposed fish (P < 0.05). The plasma glycerol concentration showed a significant decrease only in fish of 0.54 mg L-1 diazinon treatment (P < 0.05). In long term trial, the energetic metabolites (acetate, acetoacetate, glycerol) showed significant increases mostly in fish exposed to high concentrations of diazinon (P < 0.05). Phosphocreatine was detected only in groups exposed to 0.54 and 0.9 mg L-1 diazinon. Some amino acids decreased in control and diazinon-exposed groups while glycine (in control and 0.18 mg L-1 diazinon treatment), glutamine and alanine (in 0.9 mg L-1 diazinon treatment) elevated significantly after 24 h acclimation in BW (P < 0.05). Our results may help to understand the effects of pesticides on fish osmoregulation from a metabolic approach.

Boule gene expression underpins the meiotic arrest in spermatogenesis in male rainbow trout (Oncorhynchus mykiss) exposed to DEHP and butachlor.

Boule, the ancestor of the DAZ (Deleted in AZoospermia) gene family, in most organisms is mainly involved in male meiosis. The present study investigates the effects of the plasticizer DEHP (50mg/kg body weight) and herbicide butachlor (0.39mg/L) on male rainbow trout (Oncorhynchus mykiss) for a 10-day period in two independent experiments. The results showed that plasma testosterone (T) concentrations were significantly lower in fish exposed to either DEHP or butachlor compared to the control fish (P<0.05). Fish showed a significantly elevated hepatosomatic index (HSI) in the butachlor treatment (P<0.05). However, no significant difference was observed in HSI values in the DEHP treatment (P>0.05). In addition, no significant differences were found in the gonadosomatic index (GSI) in both DEHP and butachlor treatments (P>0.05). Histologically, testes of male trout in the control groups were well differentiated and filled with large numbers of cystic structures containing spermatozoa. In contrast, the testes of male trout contained mostly spermatocytes with few spermatozoa in both treated group, suggesting that DEHP and butachlor may inhibit the progression of meiosis. Also, boule gene expression was significantly lower in the testes of male trout affected by DEHP and butachlor in comparison with their control groups (P<0.05), which confirmed the meiotic arrest in affected trout. Based on the results, the present study demonstrated that DEHP and butachlor can inhibit the progression of spermatogenesis in male trout, potentially by causing an arrest of meiosis, maybe due to down-regulation of boule gene expression through T and/or IGF1 via ERK1/2 signaling in T-independent pathways. In addition, these results confirmed that boule can be considered as a predictive marker to assess meiotic efficiency.

Modulation of innate immune response, mucosal parameters and disease resistance in rainbow trout (Oncorhynchus mykiss) upon synbiotic feeding.

The present study investigates the effects of dietary supplements of galactooligosaccharides (GOS), Pediococcus acidilactici and P. acidilactici + GOS on innate immune response, skin mucus as well as disease resistance of rainbow trout (Oncorhynchus mykiss) fingerlings (15.04 ± 0.52 g). After 8 weeks of feeding, several innate immune (lysozyme, alternative complement and respiratory burst activities) and skin mucus parameters (bactericidal activity against Streptococcus faecium, Streptococcus iniae, Serratia marcescens, Staphylococcus aureus and Escherichia coli and mucus protein content) were studied. The results indicated that the three supplemented diet significantly increased innate immune response and skin mucus parameters in rainbow trout. The highest innate immune response, skin mucus activity as well as protein level was observed in synbiotic fed fish. Furthermore, at the end of the feeding experiment, some fish were intraperitoneally injected with Streptococcus iniae to determine the disease resistance. The mortality of fingerlings fed supplemented diet was significantly lower than fish from control group being the lowest mortality recorded in synbiotic fed fish group.

Effects of (Anti) Androgenic Endocrine Disruptors (DEHP and Butachlor) on Immunoglobulin M (IgM) and Leukocytes Counts of Male Rainbow Trout (Oncorhynchus mykiss).

The effect of two anti-androgenic endocrine disrupting compounds, i.e. the plasticizer di (2-ethylhexyl) phthalate (DEHP) and herbicide butachlor, were evaluated for their effects on immunoglobulin M (IgM) and leukocytes in male rainbow trout. Also, plasma testosterone (T) concentration was measured to confirm their anti-androgenic effects. In the first experiment, trout were treated with 50 mg/kg (body weight) DEHP intraperitoneally, and in the second one, fish were exposed to 0.39 mg/L butachlor for 10 days. The results showed that T concentrations and white blood cells were significantly lower in fish exposed to either DEHP or butachlor compared to control fish (p < 0.05). Fish showed significantly elevated neutrophil levels and decreased lymphocyte levels in the butachlor (p < 0.05); however, no significant difference was observed in lymphocyte and neutrophils values in the DEHP treatment (p > 0.05). In addition, no significant differences were found in IgM, eosinophil and monocyte parameters in either DEHP or butachlor treatments (p > 0.05). These results confirmed that leukocytes counts can be considered as a novel marker of immunotoxicity triggered by (anti) androgenic endocrine disruptors.

Dietary fermentable fiber upregulated immune related genes expression, increased innate immune response and resistance of rainbow trout (Oncorhynchus mykiss) against Aeromonas hydrophila.

This trial was carried out to investigate the effects of dietary administration of Vitacel(®), a commercial fermentable fiber, on immune related genes (Lysozyme, TNFα and HSP70) expression, innate immune response and resistance of rainbow trout against Aeromonas hydrophila. 120 healthy rainbow trout (81.65 ± 1.49 g) were distributed in six fiberglass tanks assigned to two treatments. The treatments were feeding rainbow trout with diets supplemented with 0 (control) or 10 g kg(-1) Vitacel(®) for 45 days. The results revealed that administration of fermentable fiber significantly (P < 0.05) upregulated lysozyme and TNFα gene expression. HSP70 gene expression was significantly lower in Vitacel(®) fed fish at the end of trial (P < 0.05). Furthermore dietary administrations of Vitacel(®) remarkably elevated rainbow trout innate immune parameters include serum lysozyme, ACH50, bactericidal activity and agglutination antibody titer (P < 0.05). Administration of 10 g kg(-1) Vitacel(®) significantly increased rainbow trout resistance against A. hydrophila (P < 0.05). The results of present study revealed that dietary Vitacel(®) can upregulates immune related genes expression and elevates innate immune response and disease resistance of rainbow trout.

The effects of dietary Immunogen on innate immune response, immune related genes expression and disease resistance of rainbow trout (Oncorhynchus mykiss).

This study investigates the effects of prebiotic Immunogen on lysozyme, TNFα and HSP70 gene expression in head kidney, humoral innate immune parameters and resistant to Aeromonas hydrophila of rainbow trout. 120 healthy rainbow trout (81.65 ± 1.49 g) were distributed in six fiberglass tanks assigned to two groups fed control or diet supplemented with 2 g kg(-1) Immunogen for 45 days. The results revealed that administration of Immunogen significantly (P < 0.05) up regulated lysozyme and TNFα gene expression. HSP70 gene expression was significantly (P < 0.05) lower in Immunogen fed fish at the end of trial. Humoral innate immune parameters (lysozyme activity, ACH50 and bactericidal activity) were significantly (P < 0.05) increased whether 15 or 45 days after feeding on Immunogen supplemented diet. However, significant (P < 0.05) increase in agglutination antibody titer observed just after 45 days feeding on Immunogen. Rainbow trout fed with 2 g kg(-1) Immunogen showed remarkably higher resistance against A. hydrophila (64.44% survival) compared to the control group (24.44% survival). These results confirm that Immunogen can up regulates immune related genes expression, stimulates immune response that per se enhances disease resistance in rainbow trout.

Effect of a glyphosate-based herbicide in Cyprinus carpio: assessment of acetylcholinesterase activity, hematological responses and serum biochemical parameters.

The objective of this study was to investigate the toxicity effects of acute and sublethal of Roundup® as a glyphosate-based herbicide on acetylcholinesterase (AChE) activity and several hematological and biochemical parameters of Cyprinus carpio. The LC50-96 h of Roundup® to C. carpio was found to be 22.19 ppm. Common carp was subjected to Roundup® at 0 (control), 3.5, 7 and 14 ppm for 16 days, and the AChE activity is verified in tissues of gill, muscle, brain and liver. After 5 days, a significant decrease was observed in the AChE activity of muscle, brain and liver tissues. Besides, a time- and dose-dependent increase in mean cell hemoglobin (MCH) and mean cell volume (MCV) was observed. In contrast, a significant decrease was found in the quantities of hemoglobin (Hb), hematocrit (HCT) and, red (RBC) and white (WBC) blood cell count. Also, the activities of aspartate aminotransferase (AST), alanine aminotransferase (ALT) and lactate dehydrogenase (LDH) in Roundup® treated groups were significantly higher than the controlled group at experimental periods. However, the level of alkaline phosphatase (ALP) had a significant reduction behavior during the sampling days. It seems that the changes in hematological and biochemical parameters as well as AChE activity could be used as efficient biomarkers in order to determine Roundup® toxicity in aquatic environment.

Optimization of recovery patterns in common carp exposed to roundup using response surface methodology: evaluation of neurotoxicity and genotoxicity effects and biochemical parameters.

The present study is the first report on optimization of recovery conditions of fishes exposed to pesticides using response surface methodology-central composite rotatable design (RSM-CCRD). The sub-lethal toxicity bioassay of Roundup® (2 ppm ~10 percent LC50, 96 h) in common carp (1, 4, 9, 16, 25, 35 and 40 day) was investigated. After exposure for 16 days to Roundup®, some the fishes were introduced to herbicide-free water. The effects of four recovery parameters including time (5-25 d), temperature (18-26 °C), water exchange rate (WER, 10-30), and salinity (0-8 ppt) on the levels of biomarkers of genotoxicity (DNA damage), neurotoxicity (acetylcholinesterase activity (AChE)), and the serum alanine (ALT) and aspartate (AST) aminotransferase in plasma were studied. The polynomial equations were significantly fitted for all response variables with high R² values (>0.95), which revealed no indication of lack of fit. The optimum conditions for the maximum AChE activity (37.14 nmol/min/mg protein) and the minimum levels of DNA damage (8.00 percent tail DNA), ALT (27.0 IU/L) and AST (91.0 IU/L) were time of 20 d, temperature of 20 °C, WER of 25 and water salinity of 6 ppt. Thus, a promising improvement for the recovery trend of fishes exposed to Roundup® stress was obtained under the optimized conditions using RSM-CCRD.

Biomarker responses in mudskipper (Periophthalmus waltoni) from the coastal areas of the Persian Gulf with oil pollution.

A suite of 5 different biomarker responses were selected and measured in 3 different phases of detoxification process in mudskipper Periophthalmus waltoni from 3 different stations including Soltani Inlet, Shif Island and Ameri Port in Bushehr coastal area of the Persian Gulf. The selected biomarkers were 7-ethoxyresorufin O-deethylase (EROD), glutathione S-transferase (GST), biliary fluorescent aromatic compounds (FACs), DNA damage using the Comet assay and micronucleus (MN). Spatial variations in biomarker responses were observed in different sites. EROD, GST, FACs, DNA damage and MN were significantly elevated in mudskippers in the most oil impacted site (Soltani inlet). Based on the assessments, bile metabolites, EROD and GST induction appear to be sensitive to polycyclic aromatic hydrocarbons (PAHs) and can be considered as complementary biomarkers; therefore P. waltoni has the potential to be used as a bioindicator in these types of ecosystems.

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.