Immunomodulatory effects of domoic acid differ between In vivo and In vitro exposure in mice

The immunotoxic potential of domoic acid (DA), a well-characterized neurotoxin, has not been fully investigated. Phagocytosis and lymphocyte proliferation were evaluated following in vitro and in vivo exposure to assay direct vs indirect effects. Mice were injected intraperitoneally with a single dose of DA (2.5 µg/g b.w.) and sampled after 12, 24, or 48 hr. In a separate experiment, leukocytes and splenocytes were exposed in vitro to 0, 1, 10, or 100 µM DA. In vivo exposure resulted in a significant increase in monocyte phagocytosis (12-hr), a significant decrease in neutrophil phagocytosis (24-hr), a significant decrease in monocyte phagocytosis (48-hr), and a significant reduction in T-cell mitogen-induced lymphocyte proliferation (24-hr). In vitro exposure significantly reduced neutrophil and monocyte phagocytosis at 1 µM. B- and T-cell mitogen-induced lymphocyte proliferation were both significantly increased at 1 and 10 µM, and significantly decreased at 100 µM. Differences between in vitro and in vivo results suggest that DA may exert its immunotoxic effects both directly and indirectly. Modulation of cytosolic calcium suggests that DA exerts its effects through ionotropic glutamate subtype surface receptors at least on monocytes. This study is the first to identify DA as an immunotoxic chemical in a mammalian species.

Effect of organophosphate, diazinon on some immunophysiological, haematological and histological variable of Rutilus frisii kutum (Kamensky,1901)

The acute toxicity and effects of diazinon on some haematological parameters of kutum (Rutilus frisii kutum, Kamensky, 1901) weighing 613.33 g±157.06 g were studied under static water quality conditions at 15°C ± 2ºC in winter and spring 2009. The effective physical and chemical parameters of water were pH= 7-8.2, dh= 300mg/L (caco3), DO= 7 ppm and T= 15°C±2ºC. The first test was primarily to determine the effects of acute toxicity (LC5096 h) of the agricultural toxicant diazinon (emulsion 60%) on kutum male brood stocks. For this purpose, 4 treatments were used to test toxicity; each treatment was repeated in 3 tanks with 9 fish per treatment and with 180 litres water capacity. After obtaining the final results, the information was analysed statistically with Probit version 1.5 (USEPA, 1985), and we determined the LC10, LC50 and LC90 values at 24 hours, 48 hours, 72 hours and 96 hours; the maximum allowable concentration value (LC5096 h divided by 10) (TRC, 1984); and the degree of toxicity. The second stage of testing consists of four treatments: LC0= 0 as experimental treatment, treatment A with a concentration of LC1= 0.107 mg/L, treatment B with concentration of LC5= 0.157 mg/L, treatment C with concentration of MAC value= 0.04 mg/L. Male brood stocks of kutum were treated with these concentrations for 45 days. Experiments were carried out under static conditions based on the standard TRC, 1984 method over 45 days. Our results show that long-term exposure to diazinon causes a decrease in the erythrocyte count (RBC), haemoglobin (Hb), haematocrit (PCV), mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH), mean corpuscular haemoglobin concentration (MCHC), leucocyte count (WBC), lymphocyte, testosterone, iron (Fe), sodium (Na), lactate dehydrogenase (LDH), and cholinesterase (CHeS). In addition, diazinon also causes an increase in prolymphocyte, aspartate aminotransferase (AST), cholesterol, alkaline phosphatase (ALP) and adrenaline (P<0.05). There are no significant effects on monocyte, eosinophil, magnesium (Mg), chloride (Cl), glucose (BS), urea (BUN), uric acid (U.A), triglyceride (TG), calcium (Ca), albumin (Alb), total protein (TP), cortisol, noradrenaline and high density lipoprotein (HDL) levels in kutum male brood stocks (P>0.05). Pathology results showed toxin diazinon no effect on average weight and fish body length, the average weight of heart, brain, spleen, liver, kidney and liver index but caueses decrease of gonad weigth and gonad index and also, cause complications of tissue necrosis, vascular congestion, inflammation in the liver, a sharp reduction in the number of glomeruli, necrosis, vascular congestion and haemorage in the kidney, capsule thickening and fibrosis, atrophy, vascular congestion, macrophages release increased, increasing sediment Hemosiderine and thickening of artery walls in the spleen, atrophy, fibrosis and necrosis in testis , vascular congestion, increased distance between the myocardium and fibrous string in heart and neuronal loss, vascular congestion and edema in the brain of kutum male brood stocks.

Comparative study of seasonal changes in some blood parameters and genetic polymorphisms of serum antibodies and antigens on red blood cells surface in immature Caspian salmon

Seasonal sampling from 40 immature Caspian salmon were performed in summer, autumn, winter and spring. The maximum ranges of RBC counts, Hct, Hb, WBC count and clotting times were observed in spring, summer, spring, spring and winter, respectively. The minimum amounts of these factors were counted in summer, winter, winter, winter and winter, respectively. Blood Samples were taken from healthy smolt, immature and adult Caspian salmon in spawning time. Hematological determinations and biochemical serum analysis were performed in 101 fish in the three samples. The ranges of hematological values for sample mean were counted. Red blood cell counts were 866600 mm3 and 1259400 mm3 in smolt and adult respectively. Hematocrit was 48.39% in smolt and 44.29% in adult. Hemoglobin was 8.85 gr/dl in smolt and 10.91 gr/dl in adult. White blood cell count was 8781.58 mm3 in smolt and 5217.55 mm3 in adult and mean were differential of WBC, Lymphocyte 90.57%in smolt and73.22% in adult. Neutrophil was 5.12% in smolt and 16.92% in adult, Monocyte were 1.27% in smolt and 4.24% in adult, Clotting time was 282.34 Seconds in smolt and 291.47 seconds in adult MCV, MCH and MCHC also meagered in smolt and adult. Biochemical parameter in immature and mature Caspian salmon meagered .Glucose concentration was 2.97 mmol.l- in immature and 1.99 mmol.l- in mature .Cholesterol concentration was 4.26 mmol.l- in immature and 7.06 mmol.l- in mature. Triglyceride amount was 2.35 mmol.l- in immature and 2.47 mmol.l- in mature and Calcium was 2.47 in immature and 2.61 mmol.l- in mature. An in situ study was made on erythrocytic isoantigens and hetero-antigen and their corresponding iso-and hetero-antibodies of sera by means of hemoagglutination tests on the blood sample, of 450 immature and 50 mature Caspian salmon. The absence of erythrocyte iso-antigens and hetero-antigen and their corresponding iso-and hetero-antibodies were shown by the experimental. It could be indicated an intra-specific variation and differences in species for kelardasht hatchery.

A comparative study on effects of Immunoster and Immunowall as immunostimulants on some hematological, biochemical and growth indices of farmed great sturgeon juveniles (Huso huso)

Prebiotics are non-digestible food ingredients that profitably affect the host by selectively stimulating the growth and /or activation of one or a limited number of bacteria in the intestine that can enhance host health status. Immunoster (IS) and Immunowall (IW) are prebiotics and immunostimulants derived from the outer cell wall of brewers yeast, Saccharomyces cerevisiae. These substances contain MOS and �-glucans. After a four-week acclimatization period to rearing conditions and basal diet, 450 farmed great sturgeon juveniles weighing 95.58 ± 9.38 g were randomly distributed into 15 fiberglass tanks (2 × 2 × 0.53 m) in five treatments (Control, IS 1%, IW 1%, IS 3%, and IW 3%) in three replicates (completely randomized design) and kept at a density of 30 fish per tank for a period of 8 weeks at water temperature 20.55 ± 5.11ºC, dissolved oxygen 6.73 ± 0.35 mg L-1 and pH 7.92 ± 0.09. IS and IW were added at two levels of 1% and 3% to the basal diet in place of cellulose, except the control. At the beginning, in the middle and at the end of the trial, carcass analysis was done to determine the moisture, protein, fat, ash, and total carbohydrate. Also, blood samples were collected to measure hematological, biochemical and immune indices. At the end of the trial, final weight, final length, body weight increase (BWI), specific growth rate (SGR), average daily growth (ADG), protein efficiency ratio (PER), feed conversion ratio (FCR), and condition factor (CF) in fish fed on IS and IW in both levels 1% and 3% showed some differences. These differences were significant in IS 3% and IW 1% and 3% compared with the control (P<0.05). HSI showed no significant difference (P>0.05) and survival rate was 100% in all treatments. Crude protein of carcass in fish fed on IS and IW at 1% and 3% showed an increase in comparison with the control. There was significant difference between IS 3% and the control in crude protein of carcass (P<0.05). Fish fed on IS and IW at 1% and 3% showed various results in hematological and biochemical factors. It was observed significant difference in MCV between IW 1% and IS 3% compared with the control (P<0.05). Although there was an increase in values of hematocrit, hemoglobin (except IS 1%), WBC (except IW 3%), MCH, neutrophil, total protein, albumin (except IS 3%), K+, and lysozyme in fish fed on IS and IW compared with the control, it was no significant (P>0.05). The maximum count of WBC and the highest value of Ca2+ were seen in IW 1%. The maximum count of lymphocyte, the highest values of total protein, albumin and IgM were recorded in IW 3%. IS 1% had the maximum count of neutrophil and the highest concentration of lysozyme. Based on obtained results, it can be declared that IS and IW at two levels of 1% and 3% can enhance growth performance and feed efficiency and also improve some hematological, biochemical, and immune indices in farmed great sturgeon juveniles.

Gross clinical signs and haematological changes associated with artifical infection of Edwardsiella tarda in Koi Carp

The occurrence of diseases is a significant setback for successful aquafarming. One of the common fish bacterial disease syndromes, Edwardsiellosis is caused by Edwardsiella tarda, a gram-negative, rod shaped bacterium associated with several diseases of marine and fresh water fish. In this study, an attempt was made to observe and analyze the onset of clinical symptoms and certain haematological parameters in Koi Carp, Cyprinus carpio L., following artificial infection with Edwardsiella tarda. The disease progress was observed and the clinical symptoms were monitored over a period of 15 days following infection. Fish were sampled at three day intervals to analyse the haematological parameters: total erythrocyte counts (RBC), total leucocyte counts (WBC), haemoglobin content and differential leucocyte count. Clinical symptoms observed included: erratic swimming behaviour, loss of appetite, haemorrhages, dropsy and exophthalmia. There was a significant decrease in the total RBC and haemoglobin levels by the 3rd and 6th day post infection, and an increase thereafter. WBC counts were higher in all infected groups in comparison to the control group. A significant increase in the number of neutrophils was found in the infected group up to the 9th day and a decrease thereafter. The lymphocyte number was significantly less up to the 12th day while the monocyte counts were significantly higher up to the 12th day post infection. The results showed that the bacterium, E. tarda, is pathogenic to Koi Carp. The hematological changes and clinical signs in infected fish reported in this paper will be helpful in the identification and the control of this infection.

The effect of vitamins C and E on some of growth factors of cultured Acipenser ruthenus

The effect of vitamins C and E on some of growth factors of cultured Acipenser ruthenus was studied in this thesis. For this purpose diets supplemented with a combination of 100 and 400 mg/kg vitamin C, L-ascorbyl-2-polyphosphate and 100 and 400 mg/kg vitamin E, D-alpha-tocopherol,were each fed to sterlet in 2 replications for 15 weeks. Fifteen fish with average weight of 350.92±14.28 gr were distributed to each of 18 tanks after adaptation with experimental diet. After 5 weeks, there were significant differences in RBC, ESR, HCT and differential counting of white blood cells among the treatments (P<0.05), but there was no significant difference in the amount of WBC among the treatments (P>0.05). After 10 weeks, there were significant differences in the amounts of Monocytes, Lymphocytes and Eosinophils (P<0.05), but there was no significant differences in the amount of other hematologic factors (P>0.05). At the end of the experiment (15th week) only WBC, RBC, Monocyte, lymphocyte and neutrophil showed significant differences between the treatments (P<0.05) and other hematologic factors did not show any significant differences between the treatments (P>0.05). The results of biochemical indices analysis showed significant differences (P<0.05) among treatments for all of the parameters for 5th weeks, total protein and glucose for 10th week and only cholesterol for 15th week. The carcass analysis at the end of experiment showed that only the amount of carbohydrate, protein and ash were significantly difference between the treatments (P<0.05). The results of growth parameters at the end of 3th, 9th, 12th and 15th week showed significant differences between the treatments (P<0.05), but at the end of 6th week only GR was significantly different between the treatments (P<0.05). After concerning acute stress test including reduce water volume, cutting the aeration for 30 minutes at the end of experiment, cortisol and glucose significantly increased (P<0.05) compared with prestress period, but the lowest response to the stressor was observed in fish fed by E400 C 400 mg/kg. On the other hand the survival was 100% during the experiment and no mortality was occurred during this period. Results of this study indicate that Vitamins C and E can have remarkable effects on hematological, biochemical and growth indices in different growth periods. So regarding the effects of vitamin C and E on some growth indices, it seems that the diet containing E400 C 100 mg/kg can be considered as the optimum diet in the rearing condition for this weigth range of fish.