Physiological and behavioral responses of land molluscs as biomarkers for pollution impact assessment: A review.

Environmental pollution caused by human activities is a global public health concern. This review discusses the impact of different types of pollutants such as pesticides, metals, nanoparticles and others on land molluscs. These molluscs are of great interest as good model organisms for studying biomarker responses in ecotoxicological monitoring programs. Several biomarkers are utilized to characterize and quantify the exposure and harmful impacts of various pollutants. In this review, we have identified physiological and behavioral endpoints (feeding, growth, avoidance, mucus secretion, locomotion and reproductive behavior) for the diagnosis of environmental pollution. The present review displays that all types of pollutants can disturb physiological and behavioral endpoints of gastropods, and these impacts depend on the matrix, exposure time and route as well as the type and concentration of the pollutants in the environment. We have also confirmed that terrestrial gastropods particularly snails as sentinel species could be used as potential bioindicator organisms for environmental quality assessment and thus for predicting potential hazards to human health.

Biomarker responses in terrestrial gastropods exposed to pollutants: A comprehensive review.

The chemical contamination of terrestrial ecosystems is a great concern as these ecosystems are the target of most of the pollutants derived from anthropogenic activities such as pesticides, heavy metals, nanoparticles, and others. Terrestrial gastropods are considered to be excellent sentinel organisms for biological monitoring of environmental pollution, as they have the ability to accumulate chemicals in their tissues and exhibit a great potential to evaluate the ecological effects of pollutants in terrestrial ecosystems. The use of biomarkers as sensitive parameters to estimate the exposure or resulting effects of chemicals have received considerable attention. The successful biomarker must be applicable in the laboratory and field conditions. Many biomarkers have been examined to understand the adverse effects of pollutants. In this review, we shed light on different types of biomarkers, such as oxidative stress, genotoxicity and immunotoxicity as diagnostic tools for monitoring the impacts of pollution. These biomarkers can provide information about early detection and quantification of these impacts during their initial manifestations and can facilitate the implementation of a rapid preventive and/or restorative responses in the affected ecosystems, as well as single or multiple biomarkers can be integrated into routine monitoring programs.

Use of multiple endpoints to investigate the ecotoxicological effects of abamectin and thiamethoxam on Theba pisana snails.

The ecotoxicological effects of dietary exposure to sublethal concentration (1/20 LC50) of abamectin (ABM) and thiamethoxam (TMX) for two weeks exposure and one week recovery on oxidative stress parameters; lipid peroxidation (LPO), reduced glutathione (GSH), catalase (CAT) and glutathione S-transferase (GST), Deoxyribonucleic acid (DNA) damage as well as immunological parameters; cell death, phagocytosis, lysosomal membrane stability (LMS), lectins, superoxide anion (O2-) generation, phenoloxidase (PO), peroxidase (POD) and hemocyanin (Hc) of the land snail, Theba pisana were examined. The results showed that both tested compounds significantly increased DNA damage, LPO content, the activity of CAT and GST, cell death, POD activity, Hc level, whereas, significantly declined in phagocytic activity, LMS, lectins level, O2- generation, PO activity, and GSH content compared to the controls after two week exposure. After one week of recovery, the measured biochemical parameters of treated snails were slightly repaired but their levels were still less than that of the level of untreated animals. Overall, based on acute toxicity endpoints, ABM appeared to be more harmful than TMX against this animal. Indeed, the different patterns of endpoint responses could represent a useful picture to characterise exposure to these pesticides in the land snail, T. pisana. This battery of snail endpoints might be a promising option to biomonitor the health of the terrestrial ecosystem and to offer valuable insights to the pesticides toxicity mechanisms.

Feeding and growth responses of the snail Theba pisana to dietary metal exposure.

The effects of dietary exposure to copper (Cu), lead (Pb), and zinc (Zn) on feeding activities, growth response, and mortality of Theba pisana snails were studied in 5-week feeding tests. Snails were fed on an artificial diet containing the following Cu, Pb, or Zn concentrations: 0, 50, 100, 500, 1000, 5000, 10,000, and 15,000 µg/g dry food. At the end of 5 weeks, the food consumption rate was decreased with increasinges in both metal concentrations and time of exposure. The estimated concentrations of metals that reduces food consumption to 50% (EC50) after 5 weeks were 56, 118, and 18 µg/g dry food for Cu, Pb, and Zn, respectively. All tested metals in the diet were found to inhibit growth of the snails in a dose-dependent manner. The toxic effect on growth of the tested metals could be arranged in the order Cu > Zn > Pb. The cumulative percentage mortality among snails fed a Cu- or Zn-contaminated diet was 73.3% and the respective value for a Pb-contaminated diet was 13.3%. There was a positive correlation between growth coefficient and food consumption for all tested metals.

Biomarkers of oxidative stress in the land snail, Theba pisana for assessing ecotoxicological effects of urban metal pollution.

This study investigates the biomarkers of oxidative stress and heavy metal accumulation for assessing ecotoxicological effects of urban metal pollution in Alexandria city, Egypt. This investigation was performed in the digestive gland of roadside Theba pisana snails obtained from six different sites in the city. Relationships between heavy metal (Zn, Cu, Pb and Cd) concentrations and oxidative stress indicators were also examined. The results showed that mean concentrations of the measured elements (mugg(-1) dry weight) were higher in polluted sites when compared to the background levels of the reference site. The pattern of metals accumulation at all sites was Zn>Cu>Pb>Cd. In the metal polluted sites, snails displayed higher mean of catalase (CAT), glutathione peroxidase (GPx) and glutathione S-transferase (GST) activities, lower reduced glutathione (GSH) content and higher levels of lipid peroxidation (LPO) compared to snails from the reference site. All oxidative stress parameters were positively correlated to heavy metal concentrations except GSH content which exhibits negative correlation with the concentrations of these metals. Our results suggest that the tested antioxidant defenses are sensitive parameters that could be useful as oxidative stress biomarkers in snails exposed to the actual metals in the environment. These biomarkers when complemented with metal accumulation in the digestive gland of snails may provide a powerful biomonitor for the assessment of environmental metal pollution.

Oxidative stress biomarkers in the digestive gland of Theba pisana exposed to heavy metals.

The in vivo toxic effects of sublethal treatment of 40 and 80% of 48-h LD(50) of topically applied trace metals [copper (Cu), lead (Pb), and zinc (Zn)] on oxidative stress biomarkers in the digestive gland of Theba pisana were examined. Oxidative individual perturbations were assessed by measuring nonenzymatic (glutathione; GSH) and enzymatic (catalase, CAT; glutathione peroxidase, GPx; and glutathione-S-transferase, GST) antioxidants in digestive gland of the snails. Lipid peroxidation (LPO) was also evaluated as a marker of cell damage. The results indicated that the copper ion was the most potent metal against this snail, followed by zinc and lead, for which the corresponding LD(50) values were 37.88, 261.72, and 652.55 microg/snail, respectively. The no-observed effect concentration (NOEC) values for Cu, Zn, and Pb were 10, 50, and 500 microg/snail, respectively, and the corresponding lowest-observed effect concentration (LOEC) values were 50, 100, and 1000 microg/snail. All trace metals resulted in a significant increase in the level of LPO, whereas a significant decline in the content of GSH was observed when compared with untreated controls. Treatment with both sublethal doses of the metals caused significant increase in CAT activity, except in the case of 40% LD(50) Zn and 80% LD(50) Cu, which exhibited no alteration in CAT when compared to control animals. GPx was significantly increased in snails exposed to 40% LD(50) Cu and Pb as well as 80% LD(50) Cu. However, an opposite effect was observed in snails exposed to 80% LD(50) Pb and in either 40 or 80% LD(50) of Zn-intoxicated animals. Treatment with Pb at two sublethal doses significantly increased GST activity, whereas treatment the animal with Cu caused significant inhibition in this enzyme. Snails exposed to 40% LD(50) Zn showed significant enhancement of GST, whereas snails exposed to 80% LD(50) showed significantly reduced GST activity. Biphasic responses were observed for CAT, GPx, and GST activities in snails exposed to Cu, Pb, and Zn, respectively. This study suggests that upregulation of the antioxidant enzyme activities, elevation of LPO, and the reduction in GSH content is related to oxidative stress in this species that could be useful as biomarkers for the evaluation of contaminated terrestrial ecosystems.

Calcified pleural plaque mimicking a traumatic pneumothorax.

Pneumothorax is a common presentation to the emergency department, arising from traumatic and non-traumatic aetiologies. Diagnosis of non-tension pneumothorax is by a visible lung edge on chest radiography together with the absence of lung markings more peripherally. The Advanced Trauma Life Support (ATLS) system recommends tube thoracostomy as the definitive treatment for traumatic pneumothorax. The case is described of a patient who presented following thoracic trauma with an unusual finding on chest radiography that led to diagnostic confusion by mimicking a pneumothorax.

In vivo evaluation of oxidative stress biomarkers in the land snail, Theba pisana exposed to copper-based pesticides.

The in vivo evaluation of oxidative stress biomarkers in the digestive gland of Theba pisana exposed to sublethal doses (40% and 80% of LD(50) after 48 h) of copper-based pesticides; copper oxychloride, copper hydroxide and copper sulphate was examined. Oxidative individual perturbations were assessed by measuring non-enzymatic (glutathione; GSH) and enzymatic (catalase; CAT, glutathione peroxidase; GPx and glutathione S-transferase; GST) antioxidants in digestive gland of the snails. Lipid peroxidation (LPO) was also evaluated as marker of cell damage. The results indicated that copper sulphate was the most potent compound against this snail followed by copper hydroxide and copper oxychloride where their corresponding LD(50) values were 26.54, 334.54 and 582.18 microg snail(-1), respectively. Copper-based compounds resulted in a significant increase in the level of LPO whereas a significant decline of GSH content in the digestive gland of snails was observed when compared with untreated controls. The CAT, GPx and GST activities of treated snails were significantly higher than those of untreated controls. In general, the activation power of these compounds was in the following order: copper sulphate > copper hydroxide > copper oxychloride. This study suggests that up-regulation of the antioxidant enzyme activities, elevation of LPO and the reduction of GSH content is related to oxidative stress in this species that they could be used as potential biomarkers of copper-based pesticides exposure.

The retrograde nail for distal femoral fractures in the elderly: high failure rate of the condyle screw and nut.

Displaced distal femoral fractures in elderly patients benefit from surgical fixation. We describe the use of a retrograde femoral nail to treat these fractures. The implant has a special condyle screw and nut that allows for manual compression of the fracture when there is an additional inter-condylar split. We present our results when using this implant to treat 16 patients over a 13-month period. The mean patient age was 78 years (range, 65-96 years). All 16 patients were available at the time of final follow up and proceeded to union at an average time of 11 weeks (range, 10-14 weeks). Twenty-six condyle screw and nuts were used. Ten out of 26 condyle screws in 10 out of 16 patients failed by disengaging at a mean time of 10 weeks (range, 8-12 weeks). Our union rate was unaffected by the nut and bolt disengaging. This would suggest that the condylar compression generated by the nut and bolt may not be critical for union.

Effect of dimethoate on the immune system of female mice.

The functional status of the immune system of female mice exposed to a single oral dose of dimethoate (16 mg/kg) was evaluated by assessing cell mediated and humoral immune responses, in addition to the effect of dimethoate on spleen and body weights after different time intervals. The data showed that dimethoate caused a time-depended decrease in spleen weights in the absence of a change in body weights. The immunologic effect of dimethoate to female mice produced a dose-dependent decrease in the number of the rosette forming cells (total and active erythrocyte rosette). The ability of splenocytes to proliferation in response to mitogens; phytohemagglutinin (PHA) for T cell and lipopolysaccharide (LPS) for B cell were significantly decreased at the different times. As compared to control, a significant decrease in serum total immunoglobulins (Ig) and IgM was found, while IgG was non-significant deceased. Results of this study also revealed that dimethoate caused a significant decrease in the number of plaque forming cell (PFC/10(6) splenocytes) in a time dependent manner.

Effects of edifenphos and glyphosate on the immune response and protein biosynthesis of bolti fish (Tilapia nilotica).

The effects of 1/1000 field recommended concentration of the organophosphorus compounds; edifenphos and glyphosate on the immune response and protein contents were investigated after different time intervals. The cell mediated immune response assessed by proliferative response of splenocytes to mitogens; phytohemagglutinin (PHA) and concanavalin A (Con A) for T cell and lipopolysaccharide (LPS) for B cell decreased significantly in tems of the level of stimulation index in the treated fish and reached maximal depression after 4 weeks. Humoral immunity assessed as splenic antibody plaque forming cells (PFC) measured after 5 days in vitro immunization to sheep erythrocytes (SRBC's) were suppressed in a concentration dependent pattern by the two compounds. The estimated ED50 for the PFC/10(6) cells of edifenphos and glyphosate were 1.48 x 10(-2) uM and 1.65 x 10(-2) uM respectively. The data also showed that serum antibody titres in the treated fish were decreased in a time dependent manner. The total protein content of serum treated with the two pesticides was decreased after different time periods compared with control. The blood serum of treated and untreated Tilapia nilotica were analyzed electrophoretically for protein components and the percentage of proteins in each fraction was determined.

Residue levels of chlorinated hydrocarbon compounds in water and sediment samples from Nile branches in the Delta, Egypt.

Water and sediment samples were collected from eight different locations along the River Nile and its branches. Residues of hexachlorocyclohexane (HCH's), hexachlorobenzene (HCB), DDT's, cyclodienes and polychlorinated biphenyls (PCB's) were analyzed by GLC. Data on Grand Total (GT) concentration values pointed out that Rosetta Branch was more polluted with all components than Demietta Branch. Kafr El-Ziate was the most polluted location showing 1355.8 ng/L for water and 7396.9 ng/g for sediments, while Delta Barrage was the least polluted site. The concentrations of gamma-HCH were higher than the other isomers (alpha- and beta-HCH) in all studied sites. The results showed that HCB was the smallest pollutant at all locations on comparison with other chlorinated hydrocarbons. El-Mansoura, Rosetta and Kafr El-Ziate sites contained the highest concentrations of DDT's in both water and sediment samples. P,P'-DDE was dominate in all locations of water samples, but P,P'-DDT was in sediment samples. Also, the results showed the prominent presence of cyclodienes when compared with the other OC's compounds in sediment samples, especially Aldrin. Kafr El-Ziate was the most polluted location by PCB's, particularly the Ar1242. However, there were increasing levels of chlorinated hydrocarbons in the sediment samples parallel to percentage extractable organic matter (% EOM). Sediment/water ratios were calculated for all locations.

Control of src kinase activity by activators, inhibitors, and substrate chaperones.

The activities of src tyrosine kinases are greatly influenced by substrate modulators (chaperones). In the presence of bovine serum albumin, the phosphorylation of a random polymer of glutamic acid, alanine, and tyrosine (1:1:1) by src kinases is stimulated 20- to 100-fold, but there is little stimulation with a polymer of glutamic acid and tyrosine (4:1) as substrate. This suggests that serum albumin interacts with the substrates rather than with the enzyme. groEL and several other heat shock proteins also stimulate the phosphorylation of a random polymer of glutamic acid, alanine, and tyrosine (1:1:1). In the absence of substrate modulators, the phosphorylation of calmodulin and of several ras proteins by src kinase is barely detectable. In the presence of polylysine or protamine, marked phosphorylation is observed. Another type of control of src kinase activities appears to be directed toward the enzyme rather than the substrate. Triton X-100 extracts of plasma membranes of bovine brain contain a heat-stable factor that stimulates c-src kinase activity with any of the polymers as substrate. The same extract contains a heat-labile factor that preferentially inhibits c-src kinase activity. The two factors are separated by DEAE-Sephacel and phosphocellulose chromatography. The presence of the activator enhances the potency of the inhibitor.

Brain protein kinase C phosphorylating poly(arginine,serine) or lamin B is stimulated by anions and by an activator purified from bovine serum albumin preparations.

The phosphorylation of histone by purified protein kinase C (PK-C) from rat brain is dependent on the presence of Ca2+ and lipids. Phosphorylation of a synthetic random polymer of arginine and serine (3:1) is only moderately enhanced by Ca2+ and lipids, but it is greatly enhanced in the absence of Ca2+ and lipids by a contaminant in crystalline bovine serum albumin or by heated cellular fractions. The phosphorylation ratio of histone to poly(arginine,serine) varies between different PK-C fractions from brains of rat, pig, or lamb. These variations are partly caused by a PK-C isozyme that prefers poly(arginine,serine) over histone as substrate. The kinase activator (KA) was partly purified from bovine serum albumin and from extracts of plasma membranes of human placenta. KA is also present in mitochondria, nuclei, and the cytosol. Sulfates and phosphates at 10 mM substitute for KA with poly(arginine,serine) as substrate. The phosphorylation of histone III in the presence of Ca2+ and lipids is moderately stimulated by KA, but the phosphorylation of lamin B and some other endogenous proteins is greatly enhanced by KA. With histones as substrates, inorganic anions do not stimulate phosphorylation. The phosphorylation of poly-(arginine,serine) is very sensitive to low concentrations of staurosporin and is inhibited by PK-C antibody, but, in contrast to histone phosphorylation, it is resistant to sphingosine and polymyxin B. The poly(arginine,serine) phosphorylating activity is more stable at 4 degrees C than the histone phosphorylating activity, but the latter is stabilized by 0.05% Triton X-100.

Assessment of lead toxicity in traffic controllers of Alexandria, Egypt, road intersections.

Blood lead level (BPbL) was determined in forty-five traffic controllers working on Alexandria road intersections. Central nervous system dysfunction in the subjects studied was investigated by means of performance tests. Biochemical indicators related to lead exposure such as delta-aminolevulinic acid dehydratase and hemoglobin in their blood were also determined. Results indicated that most of the subjects studied have a comparably high BPbL. They also showed significantly poorer performance scores than that obtained in a previous study with a group of textile workers of the same age and educational levels. The mean of the BPbL in the traffic controllers was found to be 68.28 +/- 13.22 micrograms/dl. This is a very high level compared to an acceptable level of 30.00 micrograms/dl. All neurobehavioral symptoms demonstrated in the traffic controllers could be attributed to a high level of lead exposure.

Delayed neurotoxicity in the wild mallard duckling caused by the organophosphorus insecticides cyanofenphos and leptophos.

The susceptibility of wild mallard ducklings to the delayed neurotoxic effect of the neurotoxic organophosphorus insecticides cyanofenphos and leptophos was evaluated following a daily dosing regimen. Ducklings were treated daily with either cyanofenphos or with leptophos at different dose levels for 90 days, or until they died, or became paralyzed. A control group of ducklings given corn oil at 1 ml/kg daily for 90 days was used for comparison. All treated birds were observed daily for any clinical signs of neurotoxicity during the course of this study. All of the surviving ducklings that were treated with cyanofenphos at 4 mg/kg/day or leptophos at 10 mg/kg/day developed clinical signs of delayed neurotoxicity after 7 to 11 weeks of intoxication. Symptoms included leg weakness, ataxia, severe ataxia and paralysis. The observed clinical signs were confirmed by histological changes found in the spinal cords of the treated birds. These changes were of the type associated with organophosphorus-induced delayed neuropathy (OPIDN). These results demonstrate that wild mallard ducklings are susceptible to OPIDN and this avian species can be used in screening organophosphorus compounds for such effect.