Toxicity and deleterious impacts of selenium nanoparticles at supranutritional and imbalance levels on male goldfish (Carassius auratus) sperm.

BACKGROUND: Selenium has a major role in male reproduction and antioxidative mechanisms. Although deficiency of this element can result in damages to the body's organs, this metalloid can induce deleterious effects in organisms by causing oxidative stress. This study assessed the spermatotoxicity of selenium nanoparticles (SeNPs) in goldfish (Carassius auratus) based on genotoxicity, antioxidant status, sperm quality, and histopathology. METHODS: The fish with an average weight of 70 g (n = 288) were divided into four experimental groups (three replicates) and fed three times a day with SeNPs at different levels of 0, 0.1, 0.5, and 1 mg kg diet for 30 and 60 days. RESULTS: After 30 and 60 days of feeding trial, compared to the control group, spermatocrit percentage markedly decreased at 1 mg kg SeNPs on day 30 as well as at 0.5 and 1 mg kg on day 60 (p < 0.05). Computer-assisted sperm analysis parameters especially VCL, VSL, and VAP decreased in response to SeNPs (p < 0.05). Percentage of fast speed progressive sperm cells was highest in fish fed with 0.1 mg kg SeNPs following the dietary experiment and significantly reduced in a SeNPs dose-dependent manner (p < 0.05). In addition, the levels of Malondialdehyde and Glutathione peroxidase were significantly elevated in seminal plasma of all SeNPs-treated groups (p < 0.05). On day 60, DNA damage of sperm was greatly increased at 1 mg kg SeNPs (p < 0.05). Moreover, the highest percentage of spermatocyte and spermatid were observed at the highest dose of SeNPs while the highest percentage of spermatozoa was recorded at the lowest and moderate SeNPs doses. CONCLUSION: These findings suggested that non-optimal doses of SeNPs could reduce sperm quality, induce oxidative stress, and DNA damage in sperm, and disrupt testis development.

Intra and extracellular effects of benzo [α] pyrene on liver, gill and blood of Caspian White fish (Rutilus frissi kutum): Cyto-genotoxicity and histopathology approach.

Southern parts of the Caspian Sea have been faced with a diverse range of oil pollutants. Therefore, this study was designed to evaluate the effects of relevant environmental concentrations of benzo[α]pyrene (BαP) on liver, gill, and blood of Caspian White fish. To this end, 150 fingerling fish (6.5 ± 0.8 g) were exposed to under, near and over environmental concentrations of BαP (i.e. 50, 100, and 200 ppb, respectively) and two control groups for 21 days. Following exposure to BαP, generally, DNA damage increased in the liver and gill cells as well as the frequency of micro- and bi-nucleated erythrocytes in a time and concentration-dependent pattern. In addition, the liver and gill tissues displayed several histopathological lesions. Together, the findings are warning the health status of the Caspian Sea due to an ever-increasing concentration of BαP through using Caspian White fish as an ecological model.

Immunomodulatory function of antimicrobial peptide EC-Hepcidin1 modulates the induction of inflammatory gene expression in primary cells of Caspian Trout (Salmo trutta caspius Kessler, 1877).

Hepcidins, a group of antimicrobial peptides (AMPs), play a key role in the innate immune system of fishes and act against different pathogens. In this study, antimicrobial and immune-inflammatory activity of a synthetic EC-hepcidin1, previously identified from orange-spotted grouper, were evaluated. EC-hepcidin1 showed weak activity against the zoonotic fish pathogen Streptococcus iniae (MIC 100 µg mL-1 and MBC 150 µg mL-1). To study the effect of AMPs in general, and EC-hepcidin1 in particular, a primary cell culture (SC) from the fin tissue of the Caspian Trout (Salmo trutta caspius) was established. The neutral Red method on SC cells revealed that EC-hepcidin1 has no or very low cytotoxic properties. Treatment of cells with either EC-hepcidin1 (150 µg mL-1) or fish pathogen Streptococcus iniae (MOI = 10) and a mixture of both resulted in the up-regulation of gene expression of MHC-UBA, IL-6, and TNFα indicating the modulatory function on inflammatory processes. These findings indicate that EC-hepcidin1 might act as a candidate for modulation of the innate immune system in S. iniae-based infection.

Dietary nanoencapsulated quercetin homeostated transcription of redox-status orchestrating genes in zebrafish (Danio rerio) exposed to silver nanoparticles.

The present study assessed the protective effect of chitosan-nanoencapsulated quercetin (Qu-ChiNPs) against oxidative stress caused by silver nanoparticles (AgNPs). To this end, the transcription of prime genes regulating hepatic Keap1-Nrf2 pathway as well as downstream antioxidant enzymes were monitored prior to and after oxidative stress by AgNPs. Zebrafish (Danio rerio; n = 225) was assigned into five experimental groups based on feeding with diets supplemented with different additives as follows: negative and positive control groups, without additive; ChiNPs, 400 mg nanochitosan per kg diet; Quercetin, 400 mg free quercetin per kg diet; and Qu-ChiNPs, 400 mg Qu-ChiNPs per kg diet. At the end of the feeding trial (40 days), the experimental groups, except the negative control, were exposed to sublethal concentration of AgNPs (0.15 mg L-1) for 96h. Before exposure to AgNPs, free quercetin-treated diet significantly upregulated Keap1, Nrf2, Cat, SOD, GPx, and GST genes in the liver tissue when compared with the control diet, whereas Qu-Chi.NPs downregulated their transcription to the lowest levels. After exposure to AgNPs, all genes exhibited different responses in the AgNPs-exposed groups. The highest transcription of Nrf2, Cat, SOD, GPx, and GST was observed in the positive group, with being upregulated about 8, 10, 8, 8, and 7 times, respectively, when compared to the respective ones in the negative control. However, Keap1 showed a reverse response with being transcripted 12 times lower. The quercetin treatments, especially Qu-Chi.NPs, significantly reduced the transcription of Nrf2, Cat, SOD, GPx, and GST genes, yet enhanced Keap1 expression. Qu-Chi.NPs reduced the expression of Nrf2, SOD, Cat, GPx, and GST about 11, 10, 15, 10, and 10 times, respectively, yet increased that of Keap1 about 12 times. Taken together, nanoencapsulation can improve the antioxidant efficacy of quercetin against AgNPs toxicity and might reduce involvement of the cellular antioxidant system through tuning redox status. More broadly, it would be interesting to assess the effects of Qu-Chi.NPs against other metallic and organic oxidative stressors or pollutants.

Disruptive effects of nonylphenol on reproductive hormones, antioxidant enzymes, and histology of liver, kidney and gonads in Caspian trout smolts.

The endocrine-disrupting effect of pollutants such as alkylphenols has been considered a major concern during recent years. Although the effects of nonylphenol on the reproductive hormones of fish have been investigated in several studies, the effect of this endocrine disruptor on reproductive hormones of immature fish and salmonid smolts has been less addressed. The present work studied the effects of 1, 10 and 100 µg/l concentrations of nonylphenol on the levels of plasma reproductive hormones and liver antioxidant enzymes as well as on histopathology of reproductive and non-reproductive organs of male and female Caspian brown trout (Salmo trutta caspius) smolts after 21 days of exposure. The results of the present study showed that environmentally relevant concentrations of nonylphenol affected plasma levels of sex steroids; gonadotropins, phosphorus, estradiol to testosterone ratio, and also caused histopathological lesions in liver, kidney and testis tissues of immature Caspian brown trout during smolting. Nonylphenol significantly increased the levels of estradiol in plasma of both male and female smolts exposed to nonylphenol compared with the control groups. Exposure to nonylphenol decreased testosterone and FSH levels in both genders. It has also increased plasma levels of LH in females but did not affect LH levels in male fish. Liver SOD and CAT content was decreased in nonylphenol-exposed smolts. Therefore, the release of this economically valuable and endangered species into the rivers contaminated with nonylphenol should be avoided as it can have significant effects on the development and reproductive function of smolts.

Salinity modulates biochemical and histopathological changes caused by silver nanoparticles in juvenile Persian sturgeon (Acipenser persicus).

The objective of this study was to evaluate the effect of salinity on the acute and sub-chronic toxicity of silver nanoparticles (AgNPs) in Persian sturgeon. This was evaluated by exposing Persian sturgeon to AgNPs in three salinities: freshwater (F: 0.4 ppt), brackish water 1 (B1: 6 ± 0.2 ppt), and brackish water 2 (B2: 12 ± 0.3 ppt) for 14 days, which was followed by analysis of alterations in plasma chemistry and histopathology of the gills, liver, and intestine. Values of 96-h median lethal concentration (LC50) were calculated as 0.89 mg/L in F, 2.07 mg/L in B1, and 1.59 mg/L in B2. After sub-chronic exposures, plasma cortisol, glucose, potassium, and sodium levels illustrated no significant changes within each salinity level. In F, 0.2 mg/L AgNP caused the highest levels of alkaline phosphatase and osmolality levels. In B1, 0.6 mg/L AgNP induced the highest level of alkaline phosphatase and elevated plasma osmolality was recorded in all AgNP-exposed treatments in comparison with the controls. The B2 treatment combined with 0.6 mg/L AgNP significantly reduced plasma chloride level. The results showed elevating salinity significantly increased osmolality, chloride, sodium, and potassium levels of plasma in the fish exposed to AgNPs. The abundance of the tissue lesions was AgNP concentration-dependent, where the highest number of damages was observed in the gills, followed by liver and intestine, respectively. The histopathological study also confirmed alterations such as degeneration of lamella, lifting of lamellar epithelium, hepatic vacuolation, pyknotic nuclei, and cellular infiltration of the lamina propria elicited by AgNPs in the gills, liver, and intestine of Persian sturgeon. In conclusion, the stability of AgNPs in aquatic environments can be regulated by changing the salinity, noting that AgNPs are more stable in low salinity waters.

Isolation, characterization, in vitro expansion and transplantation of Caspian trout (Salmo caspius) type a spermatogonia.

Sprmatogonial stem cells (SSCs) are valuable for preservation of endangered fish species, biological experimentation, as well as biotechnological applications. However, the rarity of SSCs in the testes has been a great obstacle in their application. Thus, establishment of an efficient in-vitro culture system to support continuous proliferation of SSCs is essential. The present study aimed to establish an efficient and simple method for in vitro culture of Caspian trout undifferentiated spermatogonial cells. Using a two-step enzymatic digestion, testicular cells were isolated from immature testes composed of mainly undifferentiated spermatogonial cells with gonadosomatic indices of <0.05%. The spermatogonial cells were purified by differential plating through serial passaging. The purified cells indicated high expression of type A spermatogonia-related genes (Ly75, Gfrα1, Nanos2, Plzf and Vasa). Proliferation of purified cells was confirmed by BrdU incorporation. Co-culture of purified cells with testicular somatic cells as a feeder layer, resulted in continuous proliferation of type A spermatogonia. The cultured cells continued to express type A spermatogonia-specific markers after one month culture. The cultured spermatogonia were successfully incorporated into the germline after being intraperitoneally transplanted into sterile triploid rainbow trout hatchlings. These results, for the first time, demonstrated that the somatic microenvironment of the rainbow trout gonad can support the colonization and survival of intraperitoneally transplanted cells derived from a fish species belonging to a different genus. Therefore, the combination of in vitro culture system and xenotransplantation can be considered as a promising strategy for conservation of Caspian trout genetic resources.

Cloning, characterization and tissue-specific expression of the antimicrobial peptide hepcidin from caspian trout (Salmo caspius) and the antibacterial activity of the synthetic peptide.

Antimicrobial peptides have a wide range of antimicrobial activity and widely occur in different organisms including mollusks, crustaceans and vertebrates. Hepcidins are a group of cysteine-rich antimicrobial peptides that are active against a variety of pathogens including gram-positive and gram-negative bacteria, as well as viruses. In this study, the hepcidin gene of Caspian trout (CtHep) was identified and characterized. Our results showed that CtHep cDNA has a 267-bp Open Reading Frame (ORF), which is translated to 88 amino acids. The CtHep was classified in the HAMP1 class of hepcidins. Comparison of DNA and cDNA sequences showed that CtHep has 3 exons and 2 introns. The signal, prodomain and mature part of CtHep have 24, 39 and 25 amino acids, respectively. The mature peptide has a molecular weight of 2881.43 Da and a theoretical isoelectric point of 8.53. The expression of CtHep mRNA was detected in different tissues of healthy and infected fish. CtHep expression in the liver, head kidney, spleen and skin was significantly enhanced after bacterial challenge. Expression of CtHep in different embryonic development stages was also substantial. Antibacterial activity of synthetic CtHep peptides was investigated against a number of Gram-positive and Gram-negative bacteria. CtHep inhibited some pathogenic bacteria such as Streptococcus iniae and Aeromonas hydrophila. In the in vivo experiment, CtHep upregulated the cytokines IL-6 and TNF-α in both kidney and spleen tissues after 24 h of the peptide injection. In conclusion, our study showed that CtHep plays an important role in the immune system of Caspian trout and also in the embryonic stages. Moreover, CtHep peptide has a potential to be used as an antimicrobial therapeutic agent as well as an immunostimulant in aquaculture.

Hematological and histopathological effects of silver nanoparticles in rainbow trout (Oncorhynchus mykiss)-how about increase of salinity?

Hematological and histopathological toxicities of silver nanoparticles (Ag-NPs) to rainbow trout were assessed in three water salinities: 0.4 ppt (low salinity), 6 ± 0.3 ppt (moderate salinity), and 12 ± 0.2 ppt (high salinity). The concentrations of Ag-NPs in the low salinity were 0.032, 0.1, 0.32, and 1 ppm, and in the moderate and high salinities were 3.2, 10, 32, and 100 ppm. The results indicated a concentration-dependently increased (thrombocyte, monocyte, and large lymphocyte) and decreased (neutrophil and small lymphocyte) white blood cell count in the Ag-NP treatments in the low salinity than the other ones in the moderate and high salinities. Red blood cell volume significantly increased in all of the experimental groups exposed to higher Ag-NP concentrations, especially those in the low salinity. In the moderate and high salinities, blood plasma total protein decreased in 10 and 32 ppm Ag-NP treatments, but albumin increased in the groups in the low salinity. Blood plasma ions (Cl-, Na+, K+, Ca2+, and Mg2+) showed high changes in the higher Ag-NP treatments. In all treatments, gill histological analysis demonstrated a time- and Ag-NP concentration-dependent extent of abnormalities, with the highest epithelial lifting in 1 ppm Ag-NPs in the low salinity and also the highest necrosis and aneurism in the 32 ppm treatments in other salinities. Lower Ag-NP concentrations in the low salinity led to fibrosis, villus fusion, inflammation, vacuolization, and microvillus hyperplasia in the gut, yet villi lifting and necrosis in 0.32 and 1 ppm of Ag-NPs were the main anomalies. In addition to the mentioned alterations, villi abolitions predominantly occurred in 32 ppm Ag-NP concentrations in the moderate and high salinities. Overall, despite exposing to lower Ag-NP concentrations, the fish kept in the low salinity demonstrated more vulnerability to Ag-NPs than those in the other salinities.

Salinity effects on osmoregulation and gill morphology in juvenile Persian sturgeon (Acipenser persicus).

The effect of abrupt and 5-day gradual salinity transfers from freshwater (FW) to 11 ‰ Caspian Sea brackish water (BW) was investigated in juvenile Persian sturgeon Acipenser persicus with three different weight groups: 1-2 g (1.62 ± 0.27 g), 2-3 g (2.55 ± 0.41 g) and 3-5 g (4.28 ± 0.76 g). Mortality rates, blood osmotic pressure, gill morphology and branchial Na+, K+-ATPase (NKA) activity were measured 4 and 10 days after abrupt transfer and 9 and 15 days after the initial gradual transfer (i.e. 4 and 10 days after reaching Caspian Sea salinity). Fish under 3 g could not survive increased salinity, and the blood osmotic pressure of the remaining surviving fish increased and remained elevated. However, heavier fish were able to survive and successfully acclimate, even to rapid salinity change with osmotic pressure reduced to Caspian Sea osmolality levels. At the gill level, the developmental increase in chloride cell volume and a higher NKA content most probably allow juveniles weighing more than 2 g to sharply increase NKA activity if the fish are transferred to BW. The rapid chloride cell proliferation occurring with increased salinity should strengthen this acclimation response. Therefore, a drastic physiological change occurs when fish weigh more than 2 g that allows migration to higher salinities. The triggering signal on chloride cells must be further investigated in order to optimize this functional step.

The response of thyroid hormones, biochemical and enzymological biomarkers to pyrene exposure in common carp (Cyprinus carpio).

Polycyclic Aromatic Hydrocarbons (PAHs) are discharged into aquatic environments through anthropogenic activities mainly industrial and municipal effluents. There is little information on the adverse effects of pyrene, a member of the PAH family which is classified as a priority pollutant by the USEPA, on fish biochemical and physiological endpoints, particularly thyroid hormones. The present study investigated the effects of subacute semi-static pyrene exposure on biochemical, enzymological and ionoregulatory responses as well as thyroid hormones in common carp (Cyprinus carpio). The fish (140±10g, 1(+) year) were exposed to 10, 50 and 100µg/l nominal concentrations of pyrene for 35 days. The results revealed that pyrene at these concentrations significantly altered plasma levels of glucose, cholesterol, triglyceride, total protein, albumin, alanine aminotransferase (ALT), aspartate aminotransferase (AST) and alkaline phosphatase (ALP). Moreover, plasma thyroid hormones (T3 and T4) were significantly decreased in fish exposed to pyrene. In contrast, plasma electrolytes (sodium, potassium and calcium) levels remained statistically unchanged after exposure to the various pyrene concentrations. In conclusion, the studied biomarkers may be used as monitoring tools to evaluate pyrene toxicity. Pyrene induced diverse effects on the physiological endpoints of common carp, thus this chemical should be considered in toxicity studies concerning PAHs. Furthermore, this study confirmed that there was an interaction between pyrene and the thyroid system in fish. Therefore, the thyroid system may be used to assess the impact of pyrene on fish.

Effects of nonylphenol on key hormonal balances and histopathology of the endangered Caspian brown trout (Salmo trutta caspius).

Endocrine disruptor chemicals (EDCs) potentially pose a hazard to endangered species. Evaluation of the sensitivity of these species to EDCs could be helpful for protecting their populations. So, the present study investigated the adverse effects of nonylphenol, an EDC, on the endocrine hormones and histopathology of male and female juvenile Caspian brown trout (Salmo trutta caspius) following 21 days of exposure to nominal concentrations of 1, 10 and 100 µg/l. The results showed that the HSI and plasma total calcium of male and female fishes exposed to 100 µg/l nonylphenol were significantly increased compared with the control groups (P<0.001). The male plasma T3 level was significantly decreased in 10 (P<0.01) and 100 (P<0.001) µg/l nonylphenol. The female T3 level increased in 1 µg/l nonylphenol concentration (P<0.05). The plasma T4 of males showed significant elevation in fishes exposed to 100 µg/l nonylphenol (P<0.05), but no change for females in any of treatment groups relative to controls (P>0.05). No significant effect of nonylphenol exposure was observed on male plasma TSH levels (P>0.05), whereas, in females, nonylphenol at all concentrations significantly reduced TSH levels. A bell-shaped response was observed in male and female plasma GH levels. Moreover, various histopathological lesions were observed in gill and intestine tissues of fishes exposed to different nonylphenol concentrations. These results demonstrate the high sensitivity of this endangered species to even environmentally relevant concentrations of nonylphenol. Furthermore, Caspian brown trout could be used as bioindicators reflecting the toxicity of nonylphenol.

Effects of short-term storage on the motility, oxidative stress, and ATP content of Persian sturgeon (Acipenser persicus) sperm.

The effective short-term storage of semen is essential when processing multiple sperm samples and when semen must be transported from collection sites to hatcheries for the fertilization of ova, or to laboratories for cryopreservation. In the present study, the spermatozoa of Persian sturgeon (Acipenser persicus) were used to evaluate the effects of short-term storage on quality parameters (the percentage of motile cells and the total period of sperm motility), oxidative stress indices, and the ATP content. Spermatozoa cells exhibited >50% motility during 6 days of storage where the average total duration of sperm motility varied from 376.42 ± 80.86 s initially to 19.28 ± 10.96 s after 6 days. No motile spermatozoa were recorded after 9 days of storage. The levels of oxidative stress indices (TBARS and CP) and antioxidant activity (SOD) increased significantly with the storage time. The ATP content also decreased significantly after 2 days of storage. The results of this study may facilitate successful reproduction management and cryopreservation protocols for this endangered fish.

Bioaccumulation of silver nanoparticles in rainbow trout (Oncorhynchus mykiss): influence of concentration and salinity.

With the increasing use of silver nanoparticles (Ag-NPs), their entrance into aquatic ecosystems is inevitable. Thus, the present study simulated the potential fate, toxicity, and bioaccumulation of Ag-NPs released into aquatic systems with different salinities. The Ag-NPs were characterized using inductively coupled plasma-atomic emission spectroscopy (ICP-AES), dynamic light scattering (DLS), transmission electron microscopy (TEM), energy-dispersive X-ray analysis (EDX), and UV-vis spectroscopy. Juvenile rainbow trout were exposed to Ag-NPs in three different salinity concentrations, including low (0.4 ppt), moderate (6 ± 0.3 ppt), and high (12 ± 0.2 ppt) salinity, for 14 days in static renewal systems. The nominal Ag-NP concentrations in the low salinity were 0.032, 0.1, 0.32, and 1 ppm, while the Ag-NP concentrations in the moderate and high salinity were 3.2, 10, 32, and 100 ppm. UV-vis spectroscopy was used during 48 h (re-dosing time) to evaluate the stability and possible changes in size of the Ag-NPs in the water. The results revealed that the λmax of the Ag-NPs remained stable (415-420 nm) at all concentrations in the low salinity with a reduction of absorbance between 380 and 550 nm. In contrast, the λmax quickly shifted to a longer wavelength and reduced absorbance in the moderate and higher salinity. The bioaccumulation of Ag in the studied tissues was concentration-dependent in all the salinities based on the following order: liver>kidneys≈gills>white muscles. All the tissue silver levels were significantly higher in the high salinity than in the moderate salinity. In addition, all the fish exposed to Ag-NPs in the low, moderate, and high salinity showed a concentration-dependent increase in their hepatosomatic index (HSI). In conclusion, most Ag-NPs that enter into freshwater ecosystems (low ionic strength) remain suspended, representing a potentially negative threat to the biota in an ionic or nanoscale form. However, in a higher salinity, nanoparticles agglomerate and precipitate on the surface of the sediment.

Comparative proteomics analysis of male and female Persian sturgeon (Acipenser persicus) gonads.

In sturgeon aquaculture, the fish are sexed by an invasive surgical examination of the gonads. Development of a non-invasive procedure for sexing fish based on a molecular method is of special interest. In the present study a proteomics approach has been utilized to analyze a differential protein expression between mature male and female Persian sturgeon (Acipenser persicus) gonads. When comparing protein patterns on the 2-DE gels of the testis and ovary, 48 unique spots were distinguished in testis while only two spots were matchless in ovary. The spots were identified by MALDI-TOF/TOF analysis. The identified proteins are involved in metabolism and energy production, cell structure, transcription and translation, cell defense, signal transduction, transport, cell division, and none were directly linked to a sex-determining gene. The provided proteomics data could be considered as a starting base for subsequent studies focusing on the identification of proteins involved in sex determination and differentiation at different stages of gonadal maturation.

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).