Mercury toxicity in two intertidal tropical marine molluscs

Lethal and sub-lethal effects of mercury have been studied in Perna viridis and Modiolus carvalhoi. For P. viridis LC30 is 1.0 p.p.m. at 48 h and 0.23 p.p.m. at 96 h. Recorded LC50 values for M. carvalhoi are 0.5 p.p.m. and 0.19 p.p.m. at 48 h and 96 h respectively. The results document that these two species, although inhabiting the same area in the tidal belt, exhibit clear differences in mercury resistance. It is further shown that the duration of exposure affects mortality rates. In sub-lethal concentration, between 0.01 and 0.10 p.p.m. decrease in pedal-gland activity is conspicuous in P. viridis. At concentrations much below LC50 values (at 96 h), although some animals are alive, pedal-gland activity is totally suspended, supporting the assumption that shell closure ability plays a minor role in byssus thread production. In M. carvalhoi total cessation of pedal gland activity occurred at 0.09 p.p.m. of mercury.

Acute toxicity of cadmium to six intertidal invertebrates

Following a static bioassay techniques the acute toxicity of cadmium to six species of intertidal invertebrates was determined. The sensitivity of the animals to cadmium was of the following order: Emerita sp. (burrowing crustacean) Donax spiculum (burrowing bivalve) Perna viridis (sedentary bivalve) Sabellaria clandestinus (tube-dwelling polychaete) Modiolus carvalhoi and Modiolus sp. (sedentary bivalves). The above observation was based on the median lethal concentrations recorded for the different species, Emerita sp. 1.35 p.p.m., Donax spiculum 1.8 p.p.m., Perna viridis 2.5 p.p.m., Sabellaria clandestinus 2.8 p.p.m., Modiolus carvalhoi 5.6 p.p.m. and Modiolus sp. 9.6 p.p.m. The findings throw insight into the toxicity of cadmium to the common intertidal animals which are either suspension or detritus feeders.

Acute toxicity of metasystox to wedge clam, Donax cuneatus from west coast

96h acute toxicity tests were performed using commercial grade metasystox on the marine wedge clam, Donax cuneatus during summer 1985. The behaviour and mortality rates were recorded periodically. Most of the dams responded in opening the shell valves and extending the siphons quicker in low test concentrations (0.004-0.0052 p.p.m) but this was slow and late in high concentrations (0.0056-0.008 p.p.m). Mortality began to occur in 0.008 p.p.m. from 12 h, whereas, in 0.0052 p.p.m. from 60 h onwards. The observed LC sub(0) value was 0.004 p.p.m. and LC sub(50) 0.0064 p.p.m. The regression equation established was Y = 79.0891 + 33.4523 X. The rate of oxygen concentration increased at LC sub(0) and LC sub(50) values compared to control indicating the disturbed physiological adjustment. The results are correlated with physico-chemical parameters of seawater and discussed in the light of pesticide toxicity to the dam.

Relationship between water, lipid and protein in the muscle, liver and gonad in a freshwater fish Puntius filamentosus (Val.)

Details are given of a study investigating the relationship between water, lipid, and protein in Puntius filamentosus. Percentage compositions in the muscle, liver and gonad of fish specimens from Lake Vellayani in India are given, showing the fluctuations which occur during maturation.

Determination of acute mercury toxicity to developing stages of Cyprinus carpio and Cirrhinus mrigala

Toxicity of inorganic mercury to different life history stages of fresh water fishes, Cyprinus carpio and Cirrhinus mrigala were demonstrated by static bioassays. 48 and 94% of egg hatching occurred in controls at 72 and 24h of experimentation in C. carpio and C. mrigala respectively. While fish eggs in water containing mercuric chloride showed delayed development as compared to the control. LC50, LC100 and safe concentrations of hatchling, fry and fingerling were calculated. Hatchling and fry were observed to be more susceptible as compared to fingerlings of C. carpio and C. mrigala.

Toxicity of malachite green to the larvae of Penaeus monodon Fabricius

A study was undertaken examining the effect of malachite green on the development and survival of the zoeae, mysis and post-larvae of Penaeus monodon. Sensitivity varied with the different larval stages; the zoeae appeared to be the least tolerant. The prophylactic potentials of malachite green in the control of Lagenidiumand Zoothamnium infesting P. monodon larvae are considered briefly. Toxicity risks may be reduced by application between ecdyses or by the removal of the dye by filtration through activated carbon.

Acute toxicity of un-ionized ammonia to milkfish (Chanos chanos) fingerlings

The acute toxicity of un-ionized ammonia to milkfish (Chanos chanos) fingerlings was determined using a static bioassay system. Median lethal concentrations found show that milkfish fingerlings have a high tolerance to ammonia and it is unlikely that levels as high as those employed for the acute exposure would be found to occur under natural conditions. Although the threat of acute toxicological effects induced by ammonia are remote, such conditions might be encountered in stressed natural environments or in heavily loaded aquaculture systems.

Acute toxicity of unionized ammonia to milkfish (Chanos chanos Forsskal) fry

The study was conducted to determine the effects of varying concentrations of ammonia to milkfish fry. Two runs of static 96h bioassays were conducted to determine the median lethal concentration (LC 50) of unionized ammonia (NH₃) to milkfish fry. Test concentrations were based on exploratory 24h and 48h bioassays and were made in three replicates. Reagent grade ammonium chloride (NH₄Cl) was used to adjust the level of unionized ammonia. The 96h median lethal concentration, determined by the Reed Muench method was calculated at 28.029 ppm NH₃ 29.69 ppm. Even at high concentrations of unionized ammonia, most of the fry mortality occurred after 48 to 96 hours exposure. Severe gill damage occurs only at concentrations above 20 ppm, especially above the LC 50. The high LC 50 value obtain shows that milkfish fry has great tolerance to ammonia, that even fry with severely-damaged gills can still recover days after it is returned to favorable culture condition. The result suggest that observed mortalities of milkfish fry under culture conditions are not due to ammonia toxicity.

Determination of bioaccumulation and microscopic survey of LAS pollutant effect on the liver, kidney and gill tissues of Rutilus frisii kutum in Caspian sea

Linear alkylbenzene sulfonate (LAS) are widely used in detergent industry. Due to contaminants entering the water, and the effects of their accumulation in fish, LAS, has a great importance in environmental pollution. In the present study, accumulation of LAS and its histological effects on gill tissue, liver and kidney of Caspian kutum (Rutilus frisii kutum) were studied. Caspian kutum is the most important and most valuable teleosts of the Caspian Sea. Due to releasing Caspian Kutum in rivers and Anzali Lagoon and unlimited entry of wastewater to the aquatic ecosystem, research on the impact of LAS on Caspian kutum is important. In the present study, fish exposed to sublethal concentrations of LAS (0.58, 1.16 and 2.32 mg/l) for 192 hours. Control treatments with three replicates at 0, 24, 48, 72, 96 and 192 hours were done. For assessments of the histological effects of LAS, tissue sections prepared and by using Hematoxylin - Eosin were stained, then the prepared sections, examined by light microscopy. For determination of the bio accumulation of LAS, the soxhlet extraction and solid phase extraction was performed to determine the amount of LAS using HPLC with fluorescence detector. According to results average of bioconcentration factor and LAS concentrations in fish had reached stable levels after approximately 72 h and thus represented steady state BCF values in this species. The value of steady-state bio-concentration factor of total LAS was 33.96 L.Kg- 1 and for each of the homologous C10-n-LAS, C11-n-LAS, C12-n-LAS and C13-n- LAS were 3.84, 6.15, 8.58 and 15.57 L.Kg-1 respectively. According to the results obtained in gills exposed to LAS, histopathological alteration include hypertrophy, lifting of lamella epithelium, edema, clubbing of lamellae hyperplasia, lamellar fusion and aneurysm were seen. In liver tissue exposed to three concentrations of LAS, congestion and dilation of sinusoids, irregular-shaped nuclei and degeneration in the hepatocyte, vacuolar degeneration and necrosis were observed. In kidney exposed to three concentrations of LAS, reduction of the interstitial haematopoietic tissue, degeneration in the epithelial cells of renal tubule, tubular degeneration, necrosis, shrinkage and luminal occlusion were observed. According to the results the most alteration due to exposure to LAS was seen in the gill tissue. None of the control samples showed histological effects of LAS.

The study of biological characteristics and heavy metal determination of Cd, Pb, Fe, V, Ni in the muscle, liver and gonad tissues of kingfish Scomberomorus commerson in the coastal waters of the Hormozagan Province (Persian Gulf)

Biological studies and heavy metal (Ni, v, Fe, Pb,Cd) determination in liver, gonad and muscle of Scomberomorus commerson were carried out from Oct 2006 to Sept 2007 in Hormozgan coastal waters. 599 Samples were gutted for reproduction and nutrition studies, fork length and weight were measured to nearest cm and g respectively. Meanwhile 40 samples were also investigated for heavy metal studies. All specimens were collected from two major landing sites (Bandar Lengeh & Bandar Abbas). Minimums & maximum fork length & weight were 29, 128 cm & 235 and 15350 g respectively.Isometric growth was shown according to our study and b was estimated 2.9 (overall), 2.91(male) & 2.89 (female). The average relative gut length was 0.52± 0.007 and it was determined that S. commerson is a carnivorous. More than 99 percent of gut content was different teleost fishes. Gastro somatic index had two peaks in Nov & Jan (before spawning) and with a decreased trend in July, the spawning period. Occurrence of empty stomach was estimated % 65.77. Maximum amount of condition factor was in Dec. Spawning season was started from June. The average of Absolute & relative fecundity (to weight unit) was 1217149±179315 and 178.2±15.58 respectively. Lm50% was estimated 75 cm for females. Sex ratio was 0.97: 1 (male: female). Chi- Square test showed no significant difference (p>0.05). Maximum amount of hepatosomatic index was estimated in March.Metal concentrations were determined using either Flame Atomic Absorption Spectroscopy (for Fe) or Graphite Furnace Atomic Absorption Spectroscopy (for Pb,Cd,Ni and V). The mean concentration (μg/g dry weight)of Pb,Cd,Ni,V and Fe in the liver were 0.0309, 0.0268, 0.0672, 0.0077, 2.5159 in the gonad 0.0440 ,0.0295, 0.1096, 0.0000, 1.4449 and in the muscle 0.0244, 0.0324, 0.0656, 0.0128, 1.6138 respectively. The maximum metal concentrations were below the maximum permissible limits for human consumption recommended by the USEPA, WHO and the UK. The results of Kendall's Tau-b correlation coefficient were as follows: The Liver tissue: There were significant positive linear relationships between accumulation of V, Fe, and Pb with Fork length, Pb and Fe with weight, GSI with Pb, Cd, V and 109 Fe, and a negative linear relationships between HSI with accumulation of V and Fe, Fork length, weight and GSI. The Gonad tissue: There were significant positive linear relationships between GSI with accumulation of Pb, Cd, Fe, Fork length and weight, a negative linear relationship between HSI with Fork length, weight and GSI. The Muscle tissue: There were significant positive linear relationships between accumulations of V, Fe with Fork length and weight factors and as well as GSI with Cd, V, Fe, Pb, Fork length and weight,a negative linear relationship between HSI with Fork length, weight, Cd, Fe and GSI. The results of Mann-Whitney U tests (P≤0.05) show that there were significant differences between summer and autumn from heavy metal contents in the studied tissues point of view. The only exceptions were for Ni in the liver, gonads and muscle and as well as there were significant differences between male and female from heavy metal contents in the studied tissues. The only exceptions were for Pb in muscle, Ni in liver, gonad and muscle, V in muscle, and Cd and Fe in gonads.

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.

Impact of phosphamidon and its metabolites on histopathology of the liver, gill and intestine of Labeo rohita

Impact of phosphamidon, an organophosphorus pesticide and its metabolites viz. dimethyl phosphoric acid and 2-chloro 2-diethyl carbamoylmethyl vinyl acid on histopathology of a common teleost, Labeo rohita was studied by exposing the fish to sub-lethal concentrations which were taken as 1/3rd of LC50 and were equal to 0.0123 ppm for phosphamidon, 0.0160 ppm for dimethyl phosphoric acid and 0.0167 ppm for 2-chloro 2-diethyl carbamoylmethyl vinyl acid respectively. The results revealed that hepatocytes in the liver were markedly swollen and exhibited hydropic degeneration. Fusion of primary lamellae and moderate congestion of blood vessels were evident in the gill. Intestine showed degeneration of mucosa and cellular infiltration in sub-mucosa. LC50 values and histopathological photomicrographs suggest that phosphamidon is more toxic as compared to dimethyl phosphoric and 2-chloro 2-diethyl carbamoylmethyl vinyl acid.

Toxicity test: Sponge toxin on tilapia fry

The research was conducted to determine the toxicity of extracts from five Philippine species of marine sponges on tilapia Oreochromis niloticus fry. It was found out that the most potent was the methanol extract of Dysidea herbacea, it kills with the least toxin and at the shortest time.

Liver cytochrome oxidase P4501A1 purification of Acipenser persicus and designing ELISA for measuring

Moon oxygenases related to cytochrome P450s are the molecular Biomarkers which have important role in Biotransformation of endogenous and exogenous compounds and catalazyin of many biological reactions. One of the important isoenzyme is cytochrome P4501A. This isoenzyme involved in metabolism of environment pollutnts such as PAHs. Because of its inducibility, it has a key tool for impact assesment of contaminants in aquatic environment. In this study, at first, that fractions containing Acipenser persicus and Huso huso isoenzyme were purified, and after that Antibodies against them were prepared. For isolation of isoenzyme fraction, Microsomes were prepared from fish liver using differential centrifugation at high speeds. microsomes were solubiized by cholat sodium and Emulgen. Extraction of this isoenzyme was done with the combinatuion of ionexchange chromatography and gelfiltration or chromatofocusing chromatography. Ion exchange chromatography and gel filtration were applied in DEAE sepharose fast flow and sephacryl S200 respectively and chromatofocusing was done at poly buffer 74 and 94 exchanger. The results of SDS-PAGE Showed that the molecular weight of isoenzyme was about 58±1 KDa. Furthermore the inmunoblotting results confirmed this subject. Isoenzyme activigy based on EROD (Ethoxyresorofin o-deethylase) reaction showed about 20-26 fold increase in enzyme activity of treated fish than control fish. The results of Elisa, Using monoclonal anti cod P4501A demonstrated the inducibility and highly elevated of its activity in treated sample more than the control fish. Mean while, the fish sample were showed the strong reaction to polyclonal antibody against beluga P4501A1 prepared in our Lab compared to monoclonal anti body.

The toxicity of four insecticides and a herbicide under tropical conditions

The toxicities of four insecticides and a herbicide to Tilapia macrochir were tested in the laboratory. The 24 hour LC50's were estimated as follows: Endrin 20% ,0.008 ppm; Lindane 5% granules, 4.6 ppmm; Synexa 50 (HCH) 50%), 5.6 ppm; Synex 25 (HCH 25%), 14.8 ppm; TOK herbicide (Nitrofen), 100% survival for 24 hours at 100 ppm. These estimates agree with results obtained by other workers elsewhere in the world. The laboratory determination of toxicity is important in estimating the direct effects of poisonous substances on fish, but other indirect effects may result from their use. These should be investigated in the field.