Histochemical Observation on Crude Oil Poisoning in the Respiratory Epithelium of Puntius sophore

The sensitivity of respiratory mucus cells to the aqueous extracts of 200… 2000 ppm of crude oil has been determined in Puntius sophore using histochemical techniques. The results indicate the mucus cells to be highly susceptible to the crude oil poisoning in both lethal and sublethal concentrations. Sublethal extracts induce excessive mucus production and form a protective covering over branchial epithelia. Lethal extracts, however, reduce the number and size of the mucus cells causing scanty mucus secretion in short-term exposure (≦4 h), but degenerate the mucus cells when the exposure period is prolonged (≧12 h).     

Toxicity of Crude Oil to the Survival of the Fresh Water Fish Puntius sophore (HAM.)

The exposure of Puntius sophore to different concentrations of crude oil (200… 4000 ppm) revealed that 4000 ppm was acutely lethal to the fish and mortality did not occur at <500 ppm at 21±1 °C for up to 15 days. The increase in the opercular frequency during exposure to crude oil has been recorded almost parallel to toxicant concentrations. The objectionable odour of the crude oil and the formation of coagulated mucous film over the body and gills were the main factors causing mortality among the experimental fish. The 24, 48, 78 and 96-h TL 50 values have been 3275, 1750, 1650 and 1450 ppm. The corresponding regression coefficients b were –1.8056, –1.5849, –1.6037 and –1.6497.     

Localization of Branchial Enzymes and their Significance in the Detection of Crude Oil Toxicity in Colisa fasciatus

The effect of crude oil on the histochemical localization of branchial enzymes CA, SDH, LDH, Alkpase and Acidpase was investigated in Colisa fasciatus. The activity of all the enzymes experimented here was found to have decreased when treated with the lethal solutions of crude oil except the LDH which showed an increasing trend throughout the experiment. A transitory increase in the CA and SDH was observed in the beginning but reached an almost normal level after 48 h in 1000 ppm. On the basis of sensitiveness towards crude oil toxicity, the enzymes have been arranged in the following order: LDH > SDH > CA > Alkpase > Acidpase. The results suggest that crude oil is capable of impairing the enzyme activity. The decreased or increased level of enzymes may be used as an indicator for assessing the toxicity of oil pollutants.     

Drilling mud-evoked hydranth shedding in the hydroid Tubularia crocea

At 100 000 ppm, the suspended particulate and liquid phases of a reference drilling mud and a used production mud significantly increase the hydranth shedding proportion in the hydroid coelenterate Tubularia crocea after 48 h. At 10 000 ppm only the liquid phase of the synthetic drilling mud significantly increased hydranth shedding. At 1000 ppm the 3% iron and calcium forms of the five lignosulphonates tested were found to be toxic. Other organic constituents of drilling muds found to be toxic included, tannic acid (100 ppm), paraformaldehyde (1 ppb), and water-soluble fractions of crude oil and No. 2 fuel oil (approximately 20 ppm). Mercury, cadmium, copper, cobalt, zinc and nickel were found toxic at 100 ppm. Exposure to 0.1−0.001 ppm mercury, 1% WSF of crude oil, and 10 ppm cadmium resulted in a significant decrease in the hydranth shedding proportion. This reduction in hydranth shedding proportion may reflect hormesis, a stimulation of growth by low-level exposure to a pollutant, but cannot be conclusively demonstrated until growth measurements are made.     

Influence of Venezuelan crude oil on lobster larvae

Emulsions of crude oil are lethal to larvae of the American Iobster at concentrations of 100 ppm and appear to have sublethal effects at concentrations down to 1 ppm.     

Survival of two species of amphipods in aqueous extracts of petroleum oils

Aqueous extracts of two petroleum oils, No. 2 fuel oil and Southern Louisiana crude, were tested on two amphipods, Gammarus muccronatus and Amphithoe valida, for survival. The oils were toxic at concentrations of 0.8 ppm (fuel oil) and 2.4 ppm (S. Louisiana crude). Mortalities increased with the concentration and length of exposure. Few or no young were produced at these and higher concentrations (breeding adults were decreasing rapidly in numbers). The amphipods are more sensitive to aqueous extracts of these oils than benthic polychaetes and shrimp, for which data are available.     

In situ impact of petrochemicals on the photosynthesis of the seagrass Zostera capricorni

We used photosynthetic activity (measured as chlorophyll a fluorescence) and photosynthetic pigment concentrations to assess the effect of pulsed exposures of aged crude oil (Champion Crude), dispersant (VDC) and an oil+dispersant mixture on the seagrass Zostera capricorni Aschers in laboratory and field experiments, using custom-made chambers. Samples were exposed for 10 h to 0.25% and 0.1% concentrations of aged crude oil and dispersant as well as mixtures of 0.25% oil+0.05% dispersant and 0.1% oil+0.02% dispersant. During this time and for the subsequent four day recovery period, the maximum and effective quantum yields of photosystem II (Fv/Fm and DeltaF/Fm' respectively) were measured. In the laboratory experiments, both values declined in response to oil exposure and remained low during the recovery period. Dispersant exposure caused a decline in both values during the recovery period, while the mixture of aged crude oil+dispersant had little impact on both quantum yields. In situ samples were less sensitive than laboratory samples, showing no photosynthetic impact due to dispersant and oil+dispersant mixture. Despite an initial decline in DeltaF/Fm', in situ oil-exposed samples recovered by the end of the experiment. Chlorophyll pigment analysis showed only limited ongoing impact in both laboratory and field situations. This study suggests that laboratory experiments may overestimate the ongoing impact of petrochemicals on seagrass whilst the dispersant VDC can reduce the impact of oil on seagrass photosynthesis.     

Effect of Parthenium Extract on the Metabolism of Young Colisa Fasciatus (BL. and SCHN.) (Pisces,Perciformes)

Parthenium hysterophorus seed extract possesses the potentiality to alter the fish metabolism. Lower concentrations (sublethal) of the seed extract stimulate the respiratory muscles, resulting in an increased rate of O₂ uptake, whereas higher concentrations (lethal) exhibit a neuro-toxic effect causing paralysis of the respiratory muscles manifested by a decreased rate of O₂ uptake, cessation of the respiratory pumps and casualty by asphyxiation. The seed extract also inhibits the activity of the enzymes SDH and carbonic anhydrase at the gas exchange sites. The log/log plotting of the O₂ uptake (mg/h) and the exposure period (2, 6, 12, 24, 72 and 168 h) showed slope values of 0.0744, 0.0778, 0.0322 and 0.0164, respectively, in four concentrations (10, 20, 25 and 30 %) of Parthenium extract.     

Lab tests on the biodegradation of chemically dispersed oil should consider the rapid dilution that occurs at sea

Most crude oils spread on open water to an average thickness as low as 0.1 mm. The application of dispersants enhances the transport of oil as small droplets into the water column, and when combined with the turbulence of 1 m waves will quickly entrain oil into the top 1 m of the water column, where it rapidly dilutes to concentrations less than 100 ppm. In less than 24 h, the dispersed oil is expected to mix into the top 10 m of the water column and be diluted to concentrations well below 10 ppm, with dilution continuing as time proceeds. Over the multiple weeks that biodegradation takes place, dispersed oil concentrations are expected to be below 1 ppm. Measurements from spills and wave basin studies support these calculations. Published laboratory studies focused on the quantification of contaminant biodegradation rates have used concentrations orders of magnitude greater than this, as it was necessary to ensure the concentrations of hydrocarbons and other chemicals were higher than the detection limits of chemical analysis. However, current analytical methods can quantify individual alkanes and PAHs (and their alkyl homologues) at ppb and ppm levels. To simulate marine biodegradation of dispersed oil at dilute concentrations commonly encountered in the field, laboratory studies should be conducted at similarly low hydrocarbon concentrations.     

New Larvicidal Acetogenin from the Red Alga Laurencia papillosa

A new C₁₅ acetogenin, cyclic enyne (12E)‐cis‐maneonene‐E ( 1 ) and (12Z)‐trans‐maneonene‐B ( 2 ), together with different secondary metabolites including sesquiterpenoid 2,10‐dibromo‐3‐chloro‐α‐chamigrene ( 3 ) and fatty acid aldehydes ( 4–8 ) were isolated from the petroleum ether extract of a red alga, identified as Laurencia papillosa, collected from the Saudi Red Sea coast. The L. papillosa crude extract and its active ingredient acetogenin ( 1 ) have been evaluated for the ability to control two main economically important pests, the confused flour beetle larvae Tribolium confusum and Culex pipiens mosquito larvae. Compound 1 showed high potential as a natural insecticide against these two pests. In the case of T. confusum larvae, the LD₅₀ values were 0.21 and 0.16% after 6 and 12 days of treatment with crude extract and acetogenin, respectively. On the other hand, the resulting LC₅₀ values of mosquito larvae bioassay were 30.7, 36.9, and 41.8 ppm after 24 h treatment against 2nd, 3rd, and 4th larval instars, respectively. Also, an increase of the insecticidal effects against mosquito larvae was recorded after extending treatment time to 48 h.     

Significance of cytochrome P450 system responses and levels of bile fluorescent aromatic compounds in marine wildlife following oil spills

The relationships among cytochrome P450 induction in marine wildlife species, levels of fluorescent aromatic compounds (FAC) in their bile, the chemical composition of the inducing compounds, the significance of the exposure pathway, and any resulting injury, as a consequence of exposure to crude oil following a spill, are reviewed. Fish collected after oil spills often show increases in cytochrome P450 system activity, cytochrome P4501A (CYP1A) and bile fluorescent aromatic compounds (FAC), that are correlated with exposure to polycyclic aromatic hydrocarbons (PAH) in the oil. There is also some evidence for increases in bile FAC and induction of cytochrome P450 in marine birds and mammals after oil spills. However, when observed, increases in these exposure indicators are transitory and generally decrease to background levels within one year after the exposure. Laboratory studies have shown induction of cytochrome P450 systems occurs after exposure of fish to crude oil in water, sediment or food. Most of the PAH found in crude oil (dominantly 2- and 3-ring PAH) are not strong inducers of cytochrome P450. Exposure to the 4-ring chrysenes or the photooxidized products of the PAH may account for the cytochrome P450 responses in fish collected from oil-spill sites. The contribution of non-spill background PAH, particularly combustion-derived (pyrogenic) PAH, to bile FAC and cytochrome P450 system responses can be confounding and needs to be considered when evaluating oil spill effects. The ubiquity of pyrogenic PAH makes it important to fully characterize all sources of PAH, including PAH from natural resources, e.g. retene, in oil spill studies. In addition, such parameters as species, sex, age, ambient temperature and season need to be taken into account. While increases in fish bile FAC and cytochrome P450 system responses, can together, be sensitive general indicators of PAH exposure after an oil spill, there is little unequivocal evidence to suggest a linkage to higher order biological effects, e.g. toxicity, lesions, reproductive failure.     

Effects of sheens associated with offshore oil and gas development on the feather microstructure of pelagic seabirds

Operational discharges of hydrocarbons from maritime activities can have major cumulative impacts on marine ecosystems. Small quantities of oil (i.e., 10 ml) results in often lethally reduced thermoregulation in seabirds. Thin sheens of oil and drilling fluids form around offshore petroleum production structures from currently permissible operational discharges of hydrocarbons. Methodology was developed to measure feather microstructure impacts (amalgamation index or AI) associated with sheen exposure. We collected feather samples from two common North Atlantic species of seabirds; Common Murres (Uria aalge) and Dovekies (Alle alle). Impacts were compared after feather exposure to crude oil and synthetic lubricant sheens of varying thicknesses. Feather weight and microstructure changed significantly for both species after exposure to thin sheens of crude oil and synthetic drilling fluids. Thus, seabirds may be impacted by thin sheens forming around offshore petroleum production facilities from discharged produced water containing currently admissible concentrations of hydrocarbons.     

Fenvalerate and Cypermethrin Induced Changes in the Haematological Parameters of Cyprinus carpio

Freshwater fish Cyprinus carpio have been exposed to sublethal concentrations of fenvalerate (0.010 ppm) and cypermethrin (0.020 ppm) for 6, 12, 24 and 48 hours. After each exposure period, certain blood parameters were studied. A decrease in red blood corpuscular (RBC) count, haemoglobin (Hb) concentration, packed cell volume (PCV) and mean corpuscular haemoglobin concentration (MCHC), on the one hand, and an increase in mean corpuscular volume (MCV) and mean corpuscular haemoglobin (MCH), on the other hand, have been observed with increasing time of exposure. The alterations observed may be due to the physiological disfunctioning of the haemopoietic tissue or may be due to the effect on the bone marrow. The effect of fenvalerate is more pronounced than that of cypermethrin.     

Acute toxicity of the water-soluble fraction of Cook Inlet crude oil to the Manila clam

Recognition of the inherent complexities of oil bioassays prompted the development of several apparatuses designed to deliver uniform solutions of the water-soluble components of crude oil and capable of system equilibration for both static and continuous-flow bioassays without the loss of the more volatile compounds and without the formation of emulsions. Clams from a relatively contaminated area exhibited greater resistance to the water-soluble fraction of Cook Inlet crude oil than specimens from a more pristine area. Behaviour was altered below lethal levels suggesting the possibility of using behavioural modifications as a concommitant criterion of toxicity with mortality. An increase in mortality with a longer exposure period and a significant rate of latent mortalities following the end of the 96-h exposure period suggest that future bioassays with bivalves should be determined over a longer exposure period.     

Development of a laboratory exposure system using marine fish to carry out realistic effect studies with produced water discharged from offshore oil production

A biotest system for environmentally realistic exposure of fish to produced water (PW) was developed and tested. Authentic PW was collected at an oil production platform in the North Sea and preserved by freezing in multiple aliquots a 25 L. After transport to the test laboratory onshore, daily PW aliquots were thawed, homogenised and administered to the test fish, Atlantic cod (Gadus morhua), in two diluted exposure concentrations, 0.1% and 0.5%, during a 15 d period, using a continuous flow-through exposure setup. Positive control groups were exposed to two crude oil treatments for comparison. Chemical analyses showed that alkylphenol (AP) and PAH concentrations in PW exposure waters were very low. Observations of significantly increased AP and PAH metabolite levels in PW exposed fish demonstrated the suitability of the biotest system for its use in biological exposure/effect studies of PW, and it also demonstrated the sensitivity of bile metabolites as PW exposure markers in fish. The relevance of the biotest system for PW effect studies and for validating modelled environmental risk estimates of PW dischargers from offshore oil production is discussed.     

Effects of toluene on the survival,respiration, and adenylate system of a marine isopod

Median times to narcotization (ET₅₀) of Cirolana borealis Lilljeborg by various levels of toluene in seawater were determined. Narcotization quickly became irreversible, and the ET₅₀ values can be regarded as estimates of median lethal times. Respiratory rate, ATP concentration, and energy charge (E.C.) were not significantly affected by sublethal toluene levels which caused behavioural disturbance. At 12.5 ppm of toluene (nominal concentration; ET₅₀:69 h) ATP dropped gradually to less than 1% and E.C. to about 50% of the control values during 8 days, after which time all the individuals were dead. At 125 ppm neither ATP concentration nor E.C. changed significantly during 24 h although the animals were inactive after about 1 h and died within 100 h. Due to the high degree of stabilization of adenine nucleotide levels and E.C. during environmental changes, these parameters do not appear useful as general indices of stress in environmental pollution studies.     

Effectiveness and toxicity of an oil dispersant in large outdoor salt water tanks

Use of the dispersant, Oilsperse 43, increased the dispersion of Venezuelan Guanipa crude oil. The resulting mixture was more homogeneous and the oil slick less viscous than in the oil tank. The dispersant appeared to retard formation of the familiar “crust” on the surface. A weathered crude oil plus dispersant mixture with an oil concentration of 250 μg/l was lethal to over 50% of the test organisms, green sea urchins, within 4 days. No mortalities occurred among urchins exposed to the crude oil treatment.     

Acute and Chronic Toxic Effects of Cadmium on some Haematological,Biochemical, and Enzymological Parameters in the Fresh Water Teleost Fish Channa punctatus Akute und chronisch toxische Wirkungen von Cadmium auf einige hämatologische,biochemische und enzymologische Parameter im Süßwasserfisch Channa punctatus

The haematological, biochemical, and enzymological alterations produced on exposure of Channa punctatus to LC₅₀ (11.2 mg/L) for 96 h and to a sublethal concentration (1.12 mg/L) of cadmium (Cd²⁺) for 15, 30, 60, and 120 days have been studied. The fish were hypoglycemic and hypolactemic. The pyruvate content of blood and liver decreased in acute and all stages of chronic exposure except for 30 days where significant increase was recorded. Depletion was noted in the total protein and glycogen content of liver and muscle, and the level of lactate in liver in the two types of exposure. The levels of muscle lactate and pyruvate increased in both exposures. The activities of hexokinase, glucose-6-phosphatase, and lactate dehydrogenase decreased in liver after 96 h and all stages of chronic exposure except 30 days. The activities of hexokinase and lactate dehydrogenase in muscle and glutamate-oxalacetate transaminase and glutamate-pyruvate transaminase in serum, liver, and muscles increased in all exposures. The activities of succinate and malate dehydrogenases in liver and muscles decreased after acute and all stages of chronic exposure. The above results suggest that glycolysis and gluconeogenesis were impaired in liver but glycolysis prevailed in muscle. Aerobic oxidation is adversely affected in metal exposed fish, while proteolysis was increased.     

Effect of lethal concentrations of copper on Mystus bleekeri (day)

In the batch test with daily exchange of medium the effect of copper ions at 2.4 mmol/l (Ca²⁺ + Mg²⁺) over 10 d of exposure is investigated. The LT₅₀ values and 95% confidence limits at 3.7, 2.1 and 1.0 ppm of Cu were 26.7 (23.6… 29.9), 47.5 (39.6… 57.0) and 101.5 (78.5… 130.7) h, respectively. The LC₅₀ values of copper in ppm were 4.05 (24 h), 1.69 (48 h), 0.829 (9.96 and 0.776 (240 h).     

Impaired osmoregulation in the shrimp Palaemon adspersus exposed to crude oil extract

Palaemon adspersus is a hyper- and hypo-osmoregulating shallow-water shrimp. For a maximum period of 34 days P. adspersus were exposed to 20, 70, 100 and 200 ppb WSF of North Sea crude oil. The osmotic concentration of the haemolymph was measured by freezing point depression once a week. The ability to maintain hyper-osmolality was significantly decreased (p<0.05) after one, two and three weeks exposure to 200, 70 and 100 ppb, respectively. No effect was observed in 20 ppb. After an accidental exposure of shrimps to 24 h acute oil pollution, they were placed in pure sea water. The ability to maintain hyper-osmolality was reduced one week after the accidental oil exposure, but was regained within three weeks.