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Dams represent large semi-closed reservoirs of pesticides and various organic and inorganic pollutants from agricultural and human activities, and their monitoring should receive special attention. This study evaluated the environmental health status of Bir Mcherga dam using zooplankton species. The dam has a capacity of 130 Mm3 and its waters are used for irrigation, water drinking supply, and fishery. Copepods and cladocerans (crustaceans) were collected in situ monthly between October and August 2012. Oxidative stress (CAT, MDA), neurotoxicity (AChE), and genotoxicity (micronucleus test) biomarkers were analyzed in two zooplankton species: Acanthocyclops robustus and Diaphanosoma mongolianum. High values of cells with a micronucleus were observed during summer. AChE activities were inhibited during early winter and summer. The high seasonal variability of CAT and MDA levels indicates that zooplankton is continuously exposed to different oxidative stresses. These results suggest that there is an obvious and continuous multi-faceted stress in Bir Mcherga reservoir and, consequently, an urgent monitoring of freshwater environments in Tunisia is needed, particularly those intended for human consumption and irrigation.

Bivalve species may be susceptible to environmental estrogenic compounds including estradiol (E2). However, they are able to biotransform the hormone quite readily and inactivate its estrogenic action. To study the long-term effects of elevated free E2 tissue levels, we transiently exceeded the biotransformation capacity of the clam Ruditapes decussatus by exposing them with high E2 concentrations (400 ng/L) and subsequently study the consequences on gametogenesis during the following reproductive cycle. Exposure to 400 ngE2/L led to a significant increase in tissue free E2 levels, which reached 10–50 ng E2Eq/gww. No deleterious effect on gonado-somatic index (GSI), condition index (CI), or ability to respond to the stress on stress test could be detected after a month of exposure, suggesting the absence of negative effects on the clam’s health. However, a marked increase in gametogenesis could be observed in both sexes during the exposure. Subsequent transplantation of the clams in the field allowed the normal development of the male clams and maturation of the gonads without any detrimental effect observed after 4 months. In contrast, in early July, all female clams formerly exposed to E2 showed lower health status, and only ovaries with atretic oocytes while all control and indigenous females were normal and mature. These results show a sex-specific effect of high E2 exposure and suggest either a direct or indirect role for E2 in R. decussatus’ reproduction.

Gene–environment interactions have long been known to play an important role in complex disease aetiology, such as nasal polyposis (NP). The present study supports the concept that DNA repair gene polymorphisms play critical roles in modifying individual susceptibility to environmental diseases. In fact, we investigated the role of polymorphisms in DNA repair genes and cadmium as risk factors for Tunisian patients with NP. To the best of our knowledge, this is the first report on the impact of combined effects of cadmium and ERCC3 7122 A>G (rs4150407), ERCC2 Lys751Gln (rs13181) and XRCC1 Arg399Gln (rs25487) genes in the susceptibility to NP disease. Significant associations between the risk of developing NP disease and ERCC2 [odds ratio (OR) = 2.0, 95 % confidence interval (CI) = 1.1–3.7, p = 0.023] and ERCC3 (OR = 2.2, 95 % CI = 1.2–4.1, p = 0.013) genotypes polymorphisms were observed. Blood concentrations of Cd in NP patients (2.2 μg/L) were significantly higher than those of controls (0.5 μg/L). A significant interaction between ERCC3 (7122 A>G) polymorphism and blood-Cd levels (for the median of blood-Cd levels: OR = 3.8, 95 % CI = 1.3–10.8, p = 0.014 and for the 75th percentiles of blood-Cd levels: OR = 2.7, 95 % CI = 1.1–7.2, p = 0.041) was found in association with the risk of NP disease. In addition, when we stratified ERCC2, ERCC3 and XRCC1 polymorphism genotypes by the median and 75th percentiles of blood-Cd levels, we found also significant interactions between ERCC2 (Lys751Gln) and ERCC3 (7122 A>G) genotypes polymorphism and this metal in association with NP disease. However, no interaction was found between XRCC1 (Arg399Gln) polymorphism genotypes and Cd in association with NP disease.

Tunisia water resources are limited. The country currently has 29 large dams, more than 1000 hill lakes, and 220 small dams which are essential for economic and social development given their contribution to irrigation, drinking water consumption, flooding protection, production of electrical energy, groundwater recharge, and industrial uses. Given the scarcity of these resources, it is crucial to be able to ensure the quality of freshwater environments, particularly those intended for human consumption. In this study, we meant to assess the health status of various freshwater ecosystems in different regions of Tunisia (north and center west) in order to detect genotoxic components in sediments and their potential effect on zooplankton (cladocerans). Sediment and cladoceran species were collected from dams, ponds, and temporary rivers in Tunisia. For each collection site, micronucleus (MN) assay was performed, in triplicates, using a pool of ten specimens of the same cladoceran species. MN occurrence in cladocerans varied from one site to another and MN frequencies varied between 0.67 and 22‰, suggesting the presence of genotoxic substances in certain sites. Sediment genotoxicity and mutagenicity were assessed using the SOS Chromotest and the Ames test. Sediment results showed that genotoxicity varies from one site to another displaying a quantitative and a qualitative variation of pollutant among the sites. These results suggest an urgent need for continuous monitoring of freshwater environments in Tunisia, particularly those intended for drinking water.

Although the pathophysiology underlying nasal polyposis (NP) formation is not fully understood, systemic, local, and environmental factors appear to contribute to NP disease development. This study aimed to explore the relationship between metal blood levels and NP risk. To the best of our knowledge, the current research represents the first scientific contribution reporting levels of Cr and As in blood of NP patients. In this context, 90 NP patients and 171 controls were recruited and blood samples were analyzed to determine the concentrations of As and Cr. Metal blood levels of As in patients (2.1 μg/L) were significantly higher than those of controls (1.2 μg/L). However, no significant difference in blood Cr levels was found between cases and controls. Arsenic blood levels of cigarette smokers were significantly higher than those of non-smokers. Environmental exposure and shisha consumption presented the most significant association with NP disease (OR = 10.1 and 14.1, respectively). High levels of blood As were significantly associated with NP disease (OR = 2.1). Cr blood levels were found to be associated with the four stages of polyps in both nasal cavities. This study found a strong association between nasal polyposis disease and As blood levels. These findings merit further investigation.

International audience; This study investigates the cytotoxicity and the genotoxicity induced by arsenic trioxide As2O3in human laryngeal SQ20B carcinoma cell line. SQ20B cells were exposed to graded concentrations of arsenic trioxide (2 and 5μM) for 48h. Comet assay and γ-H2AX foci formation were used for measuring DNA damages, flow cytometry was used to identify cell cycle alterations and apoptosis, while cell morphology was visualized using transmission electron microscopy. The results show a dose-dependent induction of DNA damages and double strand breaks, alterations in cell cycle and morphologic alterations of cells. These results prove that As2O3 is highly cytotoxic and genotoxic at the micromolar range ina human laryngeal carcinoma cell line.

This study assessed the responses of primary cultured haemocytes from the marine gastropod Haliotis tuberculata exposed to the increasing concentrations of industrial effluent (0, 0.5, 1, 10, 15 and 20%) discharged into the Tunisian coastal area. Analyses showed the presence of metals such as cadmium (Cd), chromium (Cr), copper (Cu), manganese (Mn), zinc (Zn), nickel (Ni) and lead (Pb) in the effluent. The effects of this mixture of pollutants on abalone haemocyte parameters were reflected by a significant decrease of cell viability, phagocytotic activity and reactive oxygen species (ROS) production as well as morphological and lysosomal membrane alterations. Thus, these results indicated that our primary culture system represents a suitable in vitro model for monitoring of anthropogenic contaminants in aquatic environments.

The present study is an attempt to assess the effects of contamination of several sites in the Red Sea coasts of Saudi Arabia using bivalves as a biomonitoring tool. Oxidative stress biomarkers (including reduced glutathione level (GSH), glutathione-S-transferase activity (GST), Malondialdehyde level (MDA) and Catalase activity (CAT)), neurotoxicity acetylcholinesterase activity (ACHE), and genotoxicity micronucleus rate (MN) were measured in three distinct tissues - digestive glands, gills and mantle - of specimens of the giant clam Tridacna maxima, collected from five sites in Saudi Arabian Red Sea coast (Al-Khuraybah, Al-Wajh, Yanbu, Rabigh and Thuwal). Our results demonstrated that T. maxima showed differential biomarker responses according to the nature of pollutants and human activity that affect the coast. This study can be considered as the first one using biomarkers to assess the state of the Red Sea coast in Saudi Arabia which must be followed by periodic studies for surveillance of aquatic pollution.

International audience; This study investigates cadmium effects on key messenger RNA (mRNA) expression (MT, MnSOD, CuZnSOD, CAT, ABCB1, HSP70, and CO1) by qPCR in the cockle Cerastoderma glaucum after chronic exposure to two high but environmentally relevant concentrations of CdCl2 (50 μg/L and 5 mg/L) for 12 h to 18 days. Cd accumulation measured in cockles' tissues is significantly higher in both treatment conditions compared to controls and in a dose-dependent manner. Stress on stress tests performed at different times of the experiment clearly demonstrated that exposure to both concentrations of Cd significantly affects cockle survival time in air. Important changes in gene transcription were also highlighted. In particular, MT, HSP70, CAT, and CuZnSOD seem to be relevant biomarkers of Cd exposure because (1) their mRNA levels increase upon exposure and (2) they are highly correlated to Cd accumulation in tissues. Results may be useful for control strategies and for the use of cockles as sentinel organisms.

Nasosinusal polyposis (NSP) is a chronic inflammatory disease of the nasal mucosa. Although the pathophysiology underlying NSP formation is not fully understood, environmental factors appear to be contributed the development of this disease. A case–control study of Tunisian patients was examined to assess the levels of cadmium (Cd) and nickel (Ni) in blood and reparse the association between the exposure to these metals and the risk of nasosinusal polyposis disease. Mean blood levels of Cd in patients (2.2 ± 12.8 μg/L) were significantly higher than those of controls (0.5 ± 0.7 μg/L). Levels of blood Cd were positively correlated with tobacco smoking and chewing among controls. The Cd and Ni concentrations among control (p = 0.001) and patient (p = 0.018) tobacco consumers (smoking, chewing, and shisha) were significantly higher than those nonconsumers. Additionally, Ni blood levels of patient and control smokers were significantly higher than those of nonsmokers. Cd levels in blood samples of NSP patients occupationally exposed for more than 14 years were eight times higher than that of nonexposed. Drinking water was also found to be incriminated as exposure sources. Among risk factors, shisha consumption, environmental exposure, and occupational exposure presented the most significant association with NSP disease (odds ratio (OR) = 14.1, 10.1, and 1.7, respectively). High levels of blood Cd (OR = 3.5) were strongly associated with NSP disease (p = 0.027). Ni blood levels were shown to be associated with the four stages of polyps in both nasal cavities (right and left) (p

Several studies have shown that many polymorphisms of the xenobiotic-metabolizing enzymes (XME) affect either enzymatic functions or are associated with various aspects of human health. Owing to the presence of these single nucleotide variants (SNVs), differences in detoxification capacity have been observed between many ethnicities. The aim of this investigation was to study the prevalence of four polymorphisms in XME among various ethnic groups. Attention was focused on polymorphisms of CYP2D6 (rs1058172, G>A, p.Arg365His), CYP1A1 (rs4646421, c.-26-728C>T), NAT1 (rs4921880, c.-85-1014T>A) and NAT2 (rs1208, A>G, p.Arg268Lys). These polymorphisms were analyzed in 261 healthy Tunisians individuals in comparison with different ethnic backgrounds from hapmap database. In addition, in silico functional prediction was also performed to determine the loss of function variants. Our results demonstrated that population’s origins widely affect the genetic variability of XME enzymes and Tunisians show a characteristic pattern. In silico predictions showed a deleterious effect for p.Arg268Lys substitution on CYP2D6 function, findings confirmed its key role played in cancer susceptibility. These data show that detoxification genes structures depend on the studied population. This suggests that ethnic differences impact on disease risk or response to drugs and therefore should be taken into consideration in genetic association studies focusing on XME enzymes. Our results provide the first report on these SNV in Tunisian population and could be useful for further epidemiological investigations including targeted therapy.

We have different possibilities and tools to assess the impact of pollution on marine ecosystems. The ecotoxicological approaches are based on the use of biomonitors and biomarkers. They aim to study the effect of toxic chemicals on the biological organisms especially at the population, community and ecosystem levels. The ultimate goal of ecotoxicology is to be able to predict the effects of pollution so that the most efficient and effective action to prevent or remediate any detrimental effect. In order to assess the impact of anthropogenic activities on the aquatic ecosystem and to insure compliance with regulation or guidelines, we use biomonitoring. This kind of approach is based on the use of biological responses in order to assess anthropogenic changes in the environment. Biomonitoring involves the use of indicator species such as filter feeding mollusk bivalves. These organisms tend to accumulate pollutants in their tissues without showing any apparent detrimental effect. Moreover, they could reflect the real bio available fraction of the pollutant. In order to have an early warning system predicting the pollution effects even at low levels, biomarkers were extensively studied. Some of them were validated in both field and in vivo conditions. In the present paper, the usefulness of bioindicators and biomarkers in pollution monitoring are discussed. An overview of results from case studies dealing with in situ, in vivo and transplantation experiments is presented.

The purpose of this study was to investigate the relationship between head and neck cancer (HNC) and environmental agents and polymorphisms in CYP1A1, CYP2D6, NAT1 and NAT2 metabolic enzymes genes. To the best of our knowledge, this is the first report on polymorphisms in CYP1A1 6310C>T, CYP2D6 Arg365His, NAT1 52936A>T and NAT2 Arg268Lys (NAT2*12A) genes and susceptibility to HNC in Tunisian population. We study the prevalence of these polymorphisms in 169 patients with HNC and 261 control subjects using polymerase chain reaction based methods in a Tunisian population. We detected an association between HNC and CYP1A1 6310C>T (TT) and CYP2D6 Arg365His (His/His) variant carriers (OR 1.75, P = 0.008 and OR 1.66, P = 0.016, respectively). No association was found between the polymorphisms genotypes of NAT1 52936T>A and NAT2 Arg268Lys and risk of HNC. An association between HNC and CYP1A1 (TT) genotype was found among patients with smoking (P = 0.011) and drinking habit (P = 0.009). The combinations of NAT1 (AT or AA) and NAT2 (AA) at-risk genotypes increased HNC risk (OR 4.23, P = 0.005 and OR 3.60, P = 0.048, respectively). However, the combinations of CYP1A1 (AA) and CYP2D6 (CC) genotypes decreased risk of HNC (OR 0.20; P = 0.006). Genetic polymorphisms in CYP1A1 and CYP2D6 may significantly associate with HNC in the Tunisian population. The results of this study suggest a possible gene–environment interaction for certain carcinogen metabolizing enzymes, but larger studies that fully evaluate the interaction are needed.

Department of Otorhinolaryngology, Habib Borguiba Hospital, Sfax, TunisiaAltered activity of DNA repair enzymes may be involvedin modulating cancer susceptibility and pathogenesis ofhead and neck cancer (HNC). We conducted a case–control study to test the association between threecommon single-nucleotide polymorphisms of XRCC1,ERCC2, and ERCC3 genes with HNC risk in Tunisianpatients. To the best of our knowle dge, this is the firstreport on polymorphisms in XRCC1, ERCC2, and ERCC3and susceptibility to HNC in our population. The geno-type analyses of XRCC1 Arg399Gln, ERCC2 Lys751Gln,and ERCC3 7122 A>G polymorphisms for 169 HNCpatients, and 261 controls were performed using thePCR-based restriction fragment length polymorphism.Stratification of the populations according to smokingand drinking habits and occupational exposure high-lighted the importance of tobacco, alcohol, and toxicsubstance as three risk co-factors for the development ofHNC. Our study suggests that only the XRCC1 Arg399Glnpolymorphism was associated with the risk of HNC in theTunisian population (OR = 2.04; P = 0.001). Further-more, the risk of HNC was associated with XRCC1Arg399Gln polymorphism stratified by occupationalexposure status (OR = 2.29; P = 0.024). However, nostatistically significant association was observed betweenthe risk of developing HNC and the ERCC2 Lys751Glnand ERCC3 A>G polymorphisms. These data suggest thatthe XRCC1 Arg399Gln polymorphism is associated withan increased risk of developing HNC, because it corre-lates with occupational exposure in Tunisian population.J Oral Pathol Med (2013)Keywords: ERCC2 and ERCC3 genes; head and neck cancer;occupational exposure; polymorphisms; XRCC1

Chronic exposure to heavy metals has long been recognized as being capable of increasing head and neck cancer (HNC) incidence, such as laryngeal (LC) and nasopharyngeal (NPC), among exposed human populations. The aim of the present study was to evaluate the concentrations of arsenic (As) and cadmium (Cd) in the blood of 145 patients (LC and NPC) and 351 controls in order to establish a potential relationship between these factors and the occurrence of LC and NPC. Mean blood levels of As and Cd in patients (5.67 and 3.51 μg/L, respectively) were significantly higher than those of controls (1.57 and 0.74 μg/L, respectively). The blood levels of As and Cd were mostly significantly higher than those of controls (p0.05) after controlling the other risk factors of HNC including tobacco smoking and chewing, and alcohol drinking. Cd levels in blood increase significantly with the number of occupational exposure years for patients (p0.05). However, seafood was not found to be contributing as an exposure source. Among these risk factors, smoking (30 pack years) and occupational exposure (20 years) presented the most significant association with HNC (OR=10.22 and 10.38, respectively, p0.001). Cd level in blood sample of cases that are occupationally exposed/tobacco users (smokers and chewers) were higher than that of non-occupationally exposed/nontobacco users (p0.001). The logistic regression model illustrated that HNC (LC+NPC) was significantly associated with blood levels of As (OR=2.41, p0.001) and Cd (OR=4.95, p0.001).

Chronic exposure to chromium (Cr) and nickel (Ni) has long been recognized as being capable to increase head and neck cancer (HNC) incidence among exposed human populations. This study represents the first biomonitoring of Cr and Ni exposure in Tunisia and focuses on a possible association with HNC risk. The aim of the present study was to evaluate the concentrations of Cr and Ni in the blood of HNC patients and controls. Metals blood levels of 169 HNC patients and 351 controls were determined using a Perkin-Elmer Analyst 800 Atomic Absorption Spectrometer. Mean blood levels of Cr and Ni in HNC cases (52.15 and 111.60 μg/L, respectively) were significantly higher than those of controls (37.04 and 30.50 μg/L, respectively). Cases' blood levels of Cr and Ni were significantly higher than those of controls after controlling for the other risk factors of HNC, including smoking, shisha consumption, occupational exposure, and nearby environment (P

This study focused on industrial effluent characterization and determination of its effects on the clam Ruditapes decussatus. The analysis results revealed that the effluent having acid pH and high concentrations of some chemicals, comparing to the norms, such as SO42-, NO3-, K+, Mg2+, F and Cd. The effect of the effluent on the clam R. decussatus was reflected by the LT50 determined after an exposition to different doses (5%, 10%, 20%, 30%) during 25 days and by the concentration/activity of two biomarkers: malondialdehyde (MDA) and acetylcholinesterase (AChE) determined after a contamination by effluent (1%) during 16 days. The stress on stress results showed that animals were stressed even with the lowest effluent concentration. An increase of MDA concentrations was observed in the digestive glands of exposed clams. Nevertheless, no significant variation of AChE activity was observed.

Chromosome/DNA instability could be one of the primary causes of malignant cell transformation. The objective of the present study was to evaluate the spontaneous genetic damages in exfoliated cells of buccal mucosa of head and neck cancer (HNC) by counting micronucleus (MN) and binucleated (BN) cells frequencies. MN and BN frequencies were significantly increased in HNC patients compared with controls (5.53 ± 3.09/1000 cells, 5.63 ± 2.99/1000 cells versus 2.36 ± 2.11/1000 cells, 3.09 ± 1.82/1000 cells,P<0.001). Regarding the gender and the age, the frequencies of the MN and BN were significantly higher than those of controls (P<0.01). The evaluation of the MN and BN frequencies revealed a significant increase (P<0.001) in the cases in relation to the control group after controlling the risk factors (tobacco smoking and chewing and occupational exposure) of HNC. Moreover, MN and BN frequencies were significantly increased in smokers and chewers compared with nonsmokers and nonchewers among patients (P<0.05). MN frequency was significantly (P=0.014) different between patients occupationally exposed (6.99 ± 3.40/1000 cells) and nonexposed (4.70 ± 2.48/1000 cells) among HNC group. The logistic regression model illustrated that HNC was significantly associated with frequencies of MN (OR = 8.63,P<0.0001) and BN (OR = 5.62,P=0.001). Our results suggest that increased chromosome/DNA instabilities may be associated with HNC.

Other than the established environmental risk factors associated with bladder cancer (BC), little is known about the genetic variations determining the individual susceptibility of this complex disease. This study aimed to investigate the relationship of BC with environmental agents and polymorphisms in XRCC1, ERCC2, and ERCC3 DNA repair genes and CYP1A1, CYP2D6, NAT1, and NAT2 xenobiotic metabolism genes through a hospital-based case-control study in Tunisia. The selection of the single nucleotide polymorphisms (SNPs) (rs25487, rs 13181, rs415407, rs446421, rs1058172, rs4921880, and rs1208) was performed using the dbSNP database. DNA genotyping was determined by PCR-RFLP after DNA extraction from whole blood. The risks of BC associated with every polymorphism as well as the studied environmental factors were estimated by multivariate-adjusted logistic regression using R software. In addition, gene-gene interactions were analyzed using generalized multifactor dimensionality reduction (GMDR) methods. Results showed that tobacco smoking and chewing parameters were significantly associated with BC risk. Single-gene variant analysis showed significant associations of the TT genotype of CYP1A1 and the rare GG genotype of ERCC2 with bladder cancer susceptibility (OR = 1.34, 95% CI 1.22-1.40, P

International audience; As cadmium may be involved in the etiology of head and neck cancers, we investigated in the present work, the cytotoxic and genotoxic effects of Cd on human larynx cells. SQ20B cells were exposed to 25 and 50 μM Cd for 48 and 72 h. Results showed a dose-dependent decrease in cell viability, especially after 48 h, associated with mitochondria alterations as showed by transmission electronic microscopy. Surprisingly, the flow cytometry shows that the cells treated with Cd have a normal proliferative cycle like the untreated cell especially in G1 or G2 phase of cell cycle. DNA damages were investigated by comet assay and immunofluorescence for gamma layer of the H2AX (g-H2AX) foci formation. Results show a strong induction of DNA double-strand breaks after Cd exposure. Overall, our results demonstrate the cytotoxicity and genotoxicity of Cd in human larynx cells and support the view that Cd could be an etiologic factor of head and neck cancers.

In this study, the variations of glycogen concentrations in Ruditapes decussatus from Sfax coasts (Tunisia) were described in relation to the reproductive cycle. Separate analyses were made of gonad, adductor muscle and ‘remainder’. The timing of gametogenic development and spawning was evaluated using qualitative histology associated with image analysis including (1) the estimation of the gonadal occupation index (GOI), (2) surface area occupied by reserve tissues and (3) variation in oocyte diameter. The reproductive cycle of R. decussatus exhibited partial asynchronization between sexes, the major difference being observed in the duration of the spawning period. Contrary to previous studies, we observed continuous partial spawning (e.g. 50% of ripe oocytes still subsisted at the partial spawning stage). During the gametogenic cycle, GOI varied significantly in males and in females. Most oocytes were ripe and ready to spawn when their diameter reached or exceeded 45 µm. The expansion of gonad was invers...

Detection and assessment of the impact of pollution on biological resources imply increasing research on early-warning markers such as metallothioneins (MTs) in metal exposure. In this paper, we have collated published information on the use of metallothioneins and metallothionein-like proteins (MTLPs) as biomarkers for environmental quality assessment in the Gulf of Gabès. In this area, some species of fish and bivalve were used as bioindicators of pollution. In these species, an induction of MTs/MTLPs by the essential metals such as Cu and Zn and the non-essential metals such as Cd was observed by different authors who suggest the potential use of these proteins as biomarkers. However, MT concentrations can be influenced by many biotic (sex, maturity stages, and tissues) and abiotic factors (temperature, salinity, and pH). This is essentially the case in field studies where many parameters can randomly affect MT levels, so the endogeneous regulation of MTs must be considered before using MTs as an indicator of heavy metal exposure. Moreover, the use of biomarker cannot be examined independently of the evaluation of techniques that enable its quantification. Therefore, the approach to the use of MTs/MTLP as biomarkers of exposure for an assessment of the physiological status of aquatic organisms is discussed in this paper.

Natural variation of metallothionein-like protein (MTLP) concentrations in the cockle Cerastoderma glaucum were analysed over a 1-year period in specimens collected from a relatively uncontaminated site in the Gulf of Gabes (Tunisia). Sampling was carried out from June 2004 to June 2005. At each sampling time, water temperature and salinity were measured. Levels of zinc, cadmium and copper were measured in the whole soft tissues of the cockle C. glaucum. MT-like protein and subcellular metal partitioning were measured in the digestive gland. A histological study was conducted simultaneously to follow the development of the gonad in relation to the reproductive cycle of this bivalve. Results showed a significant fluctuation in MTLP concentrations throughout the whole year, with a maximum value measured in summer and in the late winter. The relationship between MTLP and metal concentrations, abiotic (temperature and salinity) and biotic (mass of target tissues) and also maturity index (MI) factors ...

Chronic exposures to heavy metals via tobacco smoking and professional exposure may increase the risk of head and neck cancer, although the epidemiologic evidence is limited by problems of low study power and inadequate adjustment for tobacco and professional exposure use. Numerous scientific reviews have examined the association of various heavy metals exposure with respiratory cancer as well as other cancer types, but few have been published on head and neck cancer. The purpose of this paper, therefore, is to review the head and neck tract cancer-related data on exposure to heavy metals via smoking and working exposure and to study the major mechanisms underlying some toxic metals carcinogenesis.

Elevated cadmium concentrations were observed at the southern part of Sfax which received direct contamination via water discharge from the crude phosphate treatment plant and indirect contamination by the stock of phosphogypsum. In this area the ecosystem is completely destroyed therefore was necessary for sample collection to move toward the south. In order to determine a spatial distribution of pollution in this area and the consequences of this contamination on the cockles Cerastoderma glaucum, contour maps of cadmium (Cd), metallothionein like protein (MTLP), malondialdehyde (MDA) concentrations and acetylcholinesterase (AChE) activity were established. Contour mapping techniques showed elevated concentrations of Cd, MTLP and MDA in samples from Zarb El Ain toward the south and decreasing concentrations from the coast toward the east. The distribution pattern of AChE activity was opposite to the Cd concentrations. Our results showed that the area studied suffered from severe problems caused ...

The aim of this work was to detect in vivo effects of cadmium on marine organisms Cerastoderma glaucum and to determine the optimal conditions for the metallothionein-like protein (MTLP) induction in this species under laboratory conditions. The cockles Cerastoderma glaucum were exposed in vivo to different cadmium concentrations (50, 75, 100 and 150 µg L−1). Stress on stress test, cadmium, metallothionein-like protein (MTLP), malondialdehyde (MDA) and protein levels were measured after 5, 10, 15 and 20 days of metal exposure. The results showed that the CdCl2 concentrations of 50 µg L−1 for 15 days as well as 150 µg L−1 for 10 days provoked the highest MTLP induction. Nevertheless, the results obtained by the MDA, protein quantification and the stress on stress test showed that animals exposed to 50 µg L−1 for 15 days were in a better health status than those exposed to 150 µg L−1 CdCl2 for 10 days.

Cadmium (Cd) and metallothionein (MT) concentrations were analysed in the gills and digestive gland of clams Ruditapes decussatus collected from two sites of the Gulf of Gabès (Tunisia) which differ by their degree of pollution. The effect of biotic (maturity stages, protein, sex and body mass) and abiotic factors (temperature, salinity, site and season) on Cd and MT concentrations showed that in the reference site "Bordj d'Ungha" physiological changes caused by gamete development and maturity contributed more to changes in MT concentrations than the bioavailable Cd concentrations; whereas, in the contaminated site "El Hofra", MT induction responded to raised Cd bioavailabilities rather than to physiological changes. With a view to using MTs as metal exposure biomarker in monitoring program, our results show that the analysis in the digestive gland seem to be more relevant than in gills, and that males appear as the most promising candidates compared to females in the determination of this biomarker.

A Gram-positive, endospore-forming, rod-shaped and moderately halophilic bacterium was isolated from a salt-crust sample collected from Gharsa salt lake (Chott el Gharsa), Tunisia. The newly isolated bacterium, designated SA-Vb1(T), was identified based on polyphasic taxonomy including genotypic, phenotypic and chemotaxonomic characterization. Strain SA-Vb1(T) was closely related to the type strains of Virgibacillus marismortui and Virgibacillus olivae, with 16S rRNA gene sequence similarities of 99.7 and 99.4 %, respectively. However, strain SA-Vb1(T) was distinguished from these two type strains on the basis of phenotypic characteristics and DNA-DNA relatedness (29.4 and 5.1 %, respectively). The genetic relationship between strain SA-Vb1(T) and Virgibacillus pantothenticus IAM 11061(T) (the type strain of the type species) and other type strains of the genus was 96-98 % based on 16S rRNA gene sequence similarity and 18.3-22.3 % based on DNA-DNA hybridization. Biochemical analysis resulted in determination of major fatty acids iso-C(15 : 0), anteiso-C(15 : 0) and anteiso-C(17 : 0) (33.3, 29.2 and 9.8 %, respectively); phosphatidylglycerol, diphosphatidylglycerol and phosphatidylethanolamine were the main polar lipids and MK-7 was the predominant menaquinone ( approximately 100 %). The distinct characteristics demonstrated by strain SA-Vb1(T) represent properties of a novel species of the genus Virgibacillus, for which the name Virgibacillus salarius sp. nov. is proposed. The type strain is SA-Vb1(T) (=JCM 12946(T) =DSM 18441(T)).

The present study was designed to detect in situ effects of cadmium on marine organisms Cerastoderma glaucum from the gulf of Gabès (Tunisia). Six sampling stations were chosen: one site, relatively far from known local source of pollution, was considered as uncontaminated site and five sites clearly exposed to anthropogenic impact. Metallothionein like protein (MTLP) and sub-cellular metal partitioning were measured in cockles (C. glaucum) gills and digestive gland. Various biomarkers were also measured, including malondialdehyde (MDA) in the digestive gland and acetylcholinesterase activity (AChE) in the remainder. The health status of the cockles was assessed by using the condition index (CI). Significant differences between sites were noted for Cd concentrations, (CI) and also for the three studied biomarkers. Significant higher biomarkers response was measured in cockles from stations located in the northern part of the gulf, which are exposed directly to industrial and urban effluents, whereas the response of most biomarkers was minimal at the reference station. Positive and significant correlations were observed between MTLP and Cd concentrations in the digestive gland and in the gills. However, it must be noted that these correlations were more significant in the digestive gland, suggesting that compared to the gills, the digestive gland of C. glaucum is more suitable for monitoring metal pollution. The subcellular distribution of Cd showed that the soluble fraction was the major compartment for Cd storage, a pattern which is due to the role of MTLP in Cd detoxication. But at the most contaminated site (EH), cadmium in the digestive gland was preferentially accumulated in the insoluble fraction (P1) suggesting that the MTLP capacity in binding metals was not sufficient to avoid the binding of Cd to the insoluble fraction. Furthermore, the MTLP concentrations in the cockles from this site are lower than expected. So in highly polluted sites, MTLP in C. glaucum should not be used as a useful biomarker for metal pollution. The lipid peroxidation as presented by malondialdehyde levels, and MT-like protein concentrations increased in cockles exposed to cadmium contamination. We can, therefore, hypothesize that Cd could induce MTLP synthesis and MDA increase. While AChE had distinct and specific pattern showing that cadmium is not the only factor of the inhibition of cholinesterase activity. There are other polluting inputs engendering this inhibition.

Progesterone, testosterone and estradiol-17beta were measured by radio-immunoassay (RIA) in the gonads of the clam Ruditapes decussatus. The reproductive cycle was also investigated. Our study covered a period of one year, from September 2003 to August 2004. The chosen site "Kerkennah", located out of industrial effluents, belongs to the gulf of Gabès area (Tunisia). Steroids varied from 178 to 2459 pg g(-1) wet mass for progesterone, from 40 to 326 pg g(-1) wet mass for testosterone and from 10 to 235 pg g(-1) wet mass for estradiol-17beta in females. However in males, these steroids ranged from 304 to 2303 pg g(-1) wet mass for progesterone, from 81 to 381 pg g(-1) wet mass for testosterone and from 48 to 168 pg g(-1) wet mass for estradiol-17beta. The reproductive cycle of R. decussatus, investigated by histological examination of gonadic sections, showed that gametogenesis occurred from April to February in males and from April to November in females. Progesterone and testosterone increased at the end of gametogenesis in both sexes. The highest estradiol-17beta was recorded at the beginning of vitellogenesis in females. Fluctuations in the levels of sex steroids during the reproductive cycle suggest their possible role as endogenous modulators of gametogenesis in R. decussatus. Although this species is considered as gonochoristic, 0.83% of hermaphrodites were observed.

Summary The gametogenic cycle of the clam Ruditapes decussatus was investigated during one year. Samples were collected each month in two different sites located at the gulf of Gabes area. Gametogenic activity was characterized on the basis of histological analysis of the gonads. A classification and a scoring system were then proposed. According to our study, Ruditapes decussatus gametogenesis can be divided in four major steps: (1) a sexual rest (December- February), (2) starting of gametogenesis (February-March), (3) development (March-May), (4) ripe (from April onwards). This study revealed that the gonad of the clam Ruditapes decussatus was surrounded by muscular tissue which could represent the principal storage tissue. Two kinds of hermaphroditism appeared since maturity of gonads: simultaneous one showing gonadal tubules of both males and females in regionally distinct and separate zones and a “superposed” one showing a gonad consisting essentially of mature male gametes and few residual oocytes.

In order to document the potential endocrine disrupting and toxic effect of the municipal wastewater effluents discharged into the Sfax coastal area (South of Tunisia), specimens of clam R. decussatus were collected from a reference site and were in vivo exposed to treated sewage effluent for 30 days. To this end, estrogenic and androgenic activities were measured in the gills to assess potential accumulation and regulation of active compounds. After effluent exposure androgenic activity in organic extracts increased up to fivefold compared to controls and remained elevated, while estrogenic activity was not significantly affected by exposure. As a consequence, remarkable disruptions in the gametogenesis activity, glycogen content, and Vitellogenin-like protein levels in male clams were observed. A parallel analysis of heavy metals in clam tissues was determined. A significant uptake of Ni, Zn, and Pb in soft tissues of exposed clams was observed. The significant increase of malondialdehyde (MDA) concentrations as a function of exposure time implies that clams have been exposed to an oxidative stress probably due to the presence of high metal concentrations in sewage effluent. Correlation analysis has revealed a statistically significant and positive relationship between MDA levels and metal concentrations in clams' tissues. The acetylcholinesterase activity was not significantly affected by exposure. Altogether, these results showed that a short-term exposure to a mixture of chemical compounds released by the Sfax wastewater treatment plant induce adverse physiological and reproductive effects in R. decussatus. Further studies are underway in order to evaluate its long-term impacts on aquatic wildlife in the gulf of Gabes area.

International audience; This study assessed the responses of molecular biomarkers and heavy metal levels in Cerastoderma glaucum exposed for 1 week to two industrial effluents (1 %) discharged into the Tunisian coastal area, F1 and F2, produced by different units of production of a phosphate treatment plant. A significant uptake of metals (Cd, Cu, Zn, and Ni) was observed in exposed cockles compared to controls, with an uptake higher for F1 than for F2. A decrease in LT50 (stress on stress test) was also observed after an exposure to the effluent F1. Treatments resulted in different patterns of messenger RNA (mRNA) expression of the different genes tested in this report. Gene transcription monitoring performed on seven genes potentially involved in the tolerance to metal exposure showed that for both exposures, mechanisms are rapidly and synchronically settled down to prevent damage to cellular components, by (1) handling and exporting out metal ions through the up-regulation of ATP-binding cassette xenobiotic transporter (ABCB1) and metallothionein (MT), (2) increasing the mRNA expression of antioxidant enzymes (catalase (CAT), superoxide dismutases, CuZnSOD and MnSOD), (3) protecting and/or repairing proteins through the expression of heat shock protein 70 (HSP70) mRNAs, and (4) increasing ATP production (through the up-regulation of cytochrome c oxidase 1 (CO1)) to provide energy for cells to tolerate stress exposure. The tools developed may be useful both for future control strategies and for the use of the cockle C. glaucum as a sentinel species.

Among aquatic organisms suitable for biological monitoring, molluscs occupy a prominent place due to their wide geographic distribution, their abundance and accessibility in the field as well as in aquaculture. Molluscs reflect the degree of environmental contamination and are the most useful bioindicator tools. The study of modulation of immune system or immunomodulation in marine molluscs has become one of the privileged ways for evaluating the physiological effects of environmental factors. Physiological responses of molluscs to environmental stresses could be mediated by haemocytes. These cells are continually exposed to the external environment due to the open circulatory system of molluscs and are affected by pollutants. In fact, several studies showed the effects of different environmental contaminants on haemocyte functions (viability, phagocytosis, ROS production) as well as on proteins involved in cytoskeletal structure maintenance using the in vitro approaches. In ecotoxicology, in vitro approach is an alternative to animal testing due to the reduced use of experimental animals, low cost and rapid performance. Although several studies showed the importance of using in vitro cell models to determine the effects of different environmental contaminants on haemocyte parameters in marine molluscs, a few reviews highlight these effects. The main purpose of this paper is to summarize the recent data on the effect of some xenobiotics on haemocyte parameters in some mollusc species and then suggest future research prospects.

Metallothioneins (MTs), malonedialdehyde (MDA), and glycogen concentrations were determined in order to assess the health status of Ruditapes decussatus exposed to in situ contamination, showing the effects of time, site, and metal contamination on these three biomarkers.Metallothionein was positively correlated with Cd and Zn. If introduced in a multiple model, the factors site and time were shown to decrease significantly MT concentrations. MDA was inversely correlated with size of the animals and was affected by the factor time. Glycogen was inversely correlated with zinc and was affected by both site and time.This study constitutes a field-based validation of a multiparametric approach using specific and nonspecific biomarkers. Keywords: Biomarkers, Glycogen, Metallothionein, Malonedialdehyde, Metals, Ruditapes decussatus

International audience; In this study, a model based on primary cultured haemocytes from the gastropod mollusc Haliotis tuberculata was established to investigate the effects of cadmium chloride in vitro. Cells were exposed for 24 h to CdCl2 concentrations of 0, 1 and 100 μg ml−1. The effects of cadmium on haemocyte parameters were investigated using morphological, spectrophotometric and flow cytometry analysis. Results showed that cadmium has no significant effects on cell viability and phagocytotic activity under the tested conditions. However, haemocytes became more rounded after cadmium exposure, which could explain the significant decrease of cell area beginning at 1 μg ml−1 of CdCl2.

Despite growing evidence that bacteria, fungi, allergens, and superantigens play a prominent role in the pathophysiology of nasal polyps (NP), the exact cause of polyposis is still unknown. The etiology of NP is considered multifactorial. Until now, there is no information on the presence of heavy metals, such as cadmium (Cd), chromium (Cr), nickel (Ni), and arsenic (As) or of their role, in the pathogenesis of NP disease. In this study, concentrations of these four metals in tissue of NP were determined using Atomic Absorption Spectrometry. The Ni, Cr, and As levels in NP tissues were 2.1-, 3.2-, and 8.0-fold higher than those of normal mucosa (p 0.05), respectively. A strong effect of cumulative smoking as expressed in the number of pack per year (PY), Ni, As, and Cd levels in NP tissue samples of patients ever-smokers (1-20 and20 PY) are significantly higher than those of non-smokers (p = 0.006, 0.002, and0.001, respectively). The highest As concentrations among patients lived at polluted areas (1-25 and25 years) were observed in both nasal mucosa and NP tissues. The Ni and As in both nasal mucosa and NP tissues of patients occupationally exposed were significantly higher than non-exposed group. Cr and As levels were found to be associated with NP stage classification (p 0.05). This is the first report to describe an association between concentrations of metals (Cr, As, and Ni) in human NP tissues and the risk of NP disease. Tissue metal levels have increased due to smoking, environmental, and occupational exposure. Therefore, heavy metal exposure may increase the risk of NP in the Tunisian population. The considerable risk in the category of highest cumulative exposure argues for an association between heavy metals exposure and nasal polyposis risk. Future investigations with larger samples should better elucidate this association.

Bladder cancer was associated to exposure to several pollutants which can be absorbed, inhaled, or possibly ingested. We analyzed the frequency of micronuclei (MNC) and binucleated cells (BNC) in exfoliated cells of the oral mucosa of 24 bladder cancer (BC) patients and 48 controls residing in Southern Tunisia. An assessment was carried out on the incidence of MNC and BNC in 1,000 cells per individual. The data were analyzed with SPSS, using the chi-square and the Mann-Whitney U test, α = 0.05. The frequency of MN cells in BC cases was 2.5-fold higher, than in the control group (P < 0.001), while the difference for BNC between both groups was not significant. The smoking habits, age, and gender significantly influenced the MN but not the BNC alterations. The results of our study showed significantly increased frequencies of MN but not of BNC in exfoliated oral cells of BC patients associated with the smoking status, sex, and age. This study provides preliminary evidence that the frequency of MN in oral mucosa could be a predictive biomarker for cancers in parts of the body other than the upper aerodigestive tract, such as BC. Further scrupulous investigations are certainly warranted in order to implement this assay as a routine test in the planning and validation of cancer surveillance and prevention programs.

The aim of this work was to assess the relative importance of natural fluctuations in metallothionein-like protein (MTLP) levels associated with the sexual status of fish versus fluctuations due to metal exposure. In order to see fluctuations due to metal exposure, comparisons were made on the same fish species Scorpaena porcus sampled in polluted and unpolluted sites. The hermaphrodite fish Serranus scriba and Scorpaena porcus, in which sexes are separate, were compared at the unpolluted site to see fluctuations caused by the sexual status. In both species, metals and the MTLP levels were determined in the gills and liver. In these organs, Cd, Cu, and Zn distributions were examined in different fractions: the insoluble fraction (IF) and the cytosol divided into thermolabile compounds (HDF) and the heat stable compounds including MTLP. MTLP levels were higher in the liver (3.09 mg/g in S. porcus, 1.59 mg/g in S. scriba) than in gills (0.13 mg/g in S. porcus, 0.40 mg/g in S. scriba). For Scorpaena porcus, metals and MTLP levels varied with sex, whereas in Serranus scriba, which is a hermaphrodite species, inherent variations were also observed. At the polluted site, MTLP, Cd, and Cu concentrations in the gills of S. porcus increased but the supplementary metals were not associated with the heat stable compounds including MTLP. At this site, hepatic MTLP bound more metals than at the unpolluted site, but its binding capacity was not sufficient to avoid the binding of metals to the insoluble and the heat denaturable fractions. In light of these results and in spite of its hermaphrodism, it is questionable whether to consider S. scriba as a good candidate for biomonitoring based on MTLP. S. porcus could be useful for this purpose only if the MTLP capacity in binding metals is not exceeded. The MTLP could be considered as a biomarker only if it is investigated in relatively unpolluted sites.

Bladder cancer is a significant disease, the rates of which have increased over the few last years. However, its etiology remains as yet undefined. Cadmium, a widespread environmental carcinogen that has received considerable interest, presents evidence as a possible cause of bladder cancer. A literature review was conducted from the years 1984–2013 to study the accumulated evidence for cadmium as a possible cause of bladder cancer, including routes of cadmium exposure, accumulation, toxicity, carcinogenicity, and evidence from epidemiological and experimental studies. Special reference is devoted to cadmium nephrotoxicity, which illustrates how cadmium exerts its effects on the transitional epithelium of the urinary tract. Mechanisms of carcinogenesis are discussed. The effects of cadmium on gene expression in urothelial cells exposed to cadmium are also addressed. Despite different methodologies, several epidemiologic and nephrotoxicity studies of cadmium indicate that occupational exposure to cadmium is associated with increased risk of bladder cancer and provide additional evidence that cadmium is a potential toxic element in urothelial cells. In vitro studies provide further evidence that cadmium is involved in urothelial carcinogenesis. Animal studies encounter several problems such as morphology differences between species. Among the complex mechanisms of cadmium carcinogenesis, gene expression deregulation is the subject of recent studies on bladder cadmium-induced carcinogenesis. Further research, however, will be required to promise a better understanding of the mechanisms underlying cadmium carcinogenesis and to establish the precise role of cadmium in this important malignancy.

The incidence of bladder tumors has been dramatically increasing since the 1970s, possibly as a consequence of ongoing environmental pollution. Previous studies have provided some evidence of an association between cancer and exposure to carcinogenic metals. In order to examine the association between levels of toxic metals in patients with bladder tumors and controls, the amounts of arsenic, cadmium, chromium, and nickel were measured in tumoral lesions and adjacent normal part of the bladder mucosa excised for carcinoma and compared with those in the bladder mucosa of volunteer subjects operated for non-neoplastic diseases. The quantification of metals in tissue was assessed by atomic absorption spectroscopy. In tumoral tissues of the excised bladder mucosa, content of Cr and Ni was significantly low compared to that of adjacent normal tissues and control tissues while that of As and Cd in normal tissues adjacent to the tumor were significantly elevated compared to controls. Though the sample size was small, the present study shows that concentrations of metals such as Cd, Cr, As, and Ni in bladder tissue may be used as a biomarker of exposure. On the basis of the results obtained in this study, high amounts of As and Cd in adjacent normal parts of the bladders with carcinomas compared to controls would strongly suggest possible, individual or synergistic, effects of these pollutants on enzymatic systems, priming an oncogenic pathway.

The human health impact of the historic and current mining and industrial activities in Tunisia is not known. This study assessed the exposure to metals in the population of Southern Tunisia, using biomonitoring. The aim of this pilot study was to evaluate metal exposure on 350 participants living near mining and active industrial areas in the South of Tunisia. Blood specimens were analyzed for metals (Cd, Cr, As, and Ni) by Atomic Absorption Spectrometer equipped with Zeeman background correction and AS-800 auto sampler by graphite furnace and graphite tubes with integrated L'vov platform. The sample population was classified according to different age groups, sex, smoking habit, sea food and water drinking consumption, occupational exposure, amalgam fillings and place of residence. The blood As, Cd, Cr and Ni values expressed as mean ± SD were 1.56 ± 2.49, 0.74 ± 1.15, 35.04 ± 26.02 and 30.56 ± 29.96 μg/l, respectively. Blood Cd and Ni levels in smokers were 2 and 1.2 times, respectively, higher than in non-smokers. Blood Cd levels increase significantly with age (p = 0.002). As, Cd and Ni were significantly correlated with gender and age (p < 0.05). Cd level in blood samples of subjects occupationally exposed was 1.3 times higher than that of non-exposed. Blood metals were not significantly affected by amalgam fillings, place of living and sea food and drinking water consumption. This first biomonitoring study of metal exposure in the South of Tunisia reveals a substantial exposure to several metals. The pathways of exposure and health significance of these findings need to be further investigated.

The occurrence of head and neck cancer (HNC) is associated with smoking and alcohol drinking. Tobacco smoking exposes smokers to a series of carcinogenic chemicals. Cytochrome P450 enzymes (CYP450s), such asCYP1A1,CYP1B1, andCYP2D6, usually metabolize carcinogens to their inactive derivatives, but they occasionally convert the chemicals to more potent carcinogens. In addition, viaCYP450(CYP2E1) oxidase, alcohol is metabolized to acetaldehyde, a highly toxic compound, which plays an important role in carcinogenesis. Furthermore, twoN-acetyltransferase isozymes (NATs),NAT1andNAT2, are polymorphic and catalyze bothN-acetylation andO-acetylation of aromatic and heterocyclic amine carcinogens. Genetic polymorphisms are associated with a number of enzymes involved in the metabolism of carcinogens important in the induction of HNC. It has been suggested that such polymorphisms may be linked to cancer susceptibility. In this paper, we select four cytochromeP450enzymes (CYP1A1,CYP1BA1,CYP2D6, andCYP2E1), and twoN-acetyltransferase isozymes (NAT1andNAT2) in order to summarize and analyze findings from the literature related to HNC risk by focusing on (i) the interaction between these genes and the environment, (ii) the impact of genetic defect on protein activity and/or expression, and (iii) the eventual involvement of race in such associations.

Three fish species ( Diplodus annularis, Scorpaena porcus and Scorpaena scrofa ) were sampled off the coast of the industrialized area of Sfax. The distribution of Cd, Cu and Zn was examined in different fractions of both gills and liver: insoluble fraction and cytosol divided into thermolabile compounds and heat-stable compounds. In the latest fraction, quantitative analysis of metallothionein-like proteins (MTLP) was achieved by using differential pulse polarography. In the gills, the insoluble fraction was the major compartment for the storage of all three metals. In the liver, 50% Cd and Cu at least were also associated with this fraction whereas Zn was bound primarily with heat-stable compounds, a pattern which is probably due to the role of MTLP in Zn homeostasis and to the fact that Zn is a constituent element of metallothionein. According to the species, one eighth to a little more than one quarter of Cd, Cu or Zn was bound to the thermolabile fraction in the liver, probably due to the presence of numerous enzymes involved in the metabolic functions of this organ. In both gills and liver, Cd and Cu concentrations in the thermolabile fraction were increased with increasing organ burden of these metals. This observation suggests an enhancement of the toxic hazard since this fraction includes metabolic targets of metals. Statistically significant relationships have been shown between metals and MTLP concentrations in both organs. However, the use of MTLP as a biomarker has to be revised in the light of natural changes (according to age, sex, etc.) which could conceal variations due to metabolic pollution.

Genetic polymorphisms in some DNA repair proteins are associated with a number of malignant transformations like head and neck squamous cell carcinoma (HNSCC). Xeroderma pigmentosum group D (XPD) and X-ray repair cross-complementing proteins 1 (XRCC1) and 3 (XRCC3) genes are involved in DNA repair and were found to be associated with HNSCC in numerous studies. To establish our overall understanding of possible relationships between DNA repair gene polymorphisms and development of HNSCC, we surveyed the literature on epidemiological studies that assessed potential associations with HNSCC risk in terms of gene–environment interactions, genotype-induced functional defects in enzyme activity and/or protein expression, and the influence of ethnic origin on these associations. We conclude that large, well-designed studies of common polymorphisms in DNA repair genes are needed. Such studies may benefit from analysis of multiple genes or polymorphisms and from the consideration of relevant exposures that may influence the likelihood of HNSCC when DNA repair capacity is reduced.

Although exposure to high levels of arsenic is associated with excess bladder cancer risk, lower exposures generally are not. This study represents the first biomonitoring of arsenic exposure in Tunisia and focuses on a possible association with bladder cancer risk. In this context, 124 male bladder cancer cases and 220 controls were recruited and blood samples were analyzed to determine the concentration of As. The study subjects were stratified into median groups based on concentrations of arsenic in their blood. Blood arsenic (B-As) was significantly two to threefold higher in bladder cancer cases than in controls (p

Chronic exposure to heavy metals has long been recognized as being capable to increase head and neck cancer incidence among exposed human populations. Head and neck cancer is a significant public health issue in Tunisia. The aim of the present study was to evaluate the concentrations of As, Cd, Cr and Ni in healthy and tumor tissues of head and neck cancer patients. Metal concentrations were determined in tumor and healthy tissues of 101 head and neck cancer patients, using Atomic Absorption Spectrometry. The As, Cd, Cr, and Ni levels in tumor tissues were 3.4, 2.5, 1.3 and 1.5 times higher than those of healthy tissues (p0.05), respectively. Tumor tissue metal levels were higher in men than in women. As and Cd levels in tumor and healthy tissue samples of patients smokers are significantly higher than those of non-smokers (p0.05). A strong effect of cumulative smoking as expressed in the number of pack per year, and tumor tissue Cd levels were positively associated with three groups of age (40, 51-60 and60 years) in both never-smokers and ever-smokers (20 and ≥20 pack per year). Healthy tissue Cd levels were negatively associated with age in those three groups of smokers. The highest Cd and Cr concentrations among both workers and non-workers were observed in tumor tissues. The Cd and Cr in tissues of farmers, bricklayers and painters were all significantly higher among the workers as compared with the non-workers group. Tissue metal levels have increased due to smoking and occupational exposure. Heavy metal exposure via tobacco smoking and occupational exposures may increase the risk of head and neck in the Tunisian population.

The internal transcribed spacer 1 (ITS1) region of 29 individuals of Cerastoderma glaucum (from the Mediterranean Sea, the Atlantic and the North Sea) and 18 individuals of Cerastoderma edule (from the Atlantic and the North Sea) and the cytochrome c oxidase subunit I (COI) region of 43 individuals of C. glaucum (from the Mediterranean Sea, the Atlantic and the North Sea) and 9 individuals of C. edule (from the Atlantic and the North Sea) were PCR amplified and sequenced. The ITS1 sequences led us to describe 27 genetic haplotypes, while 21 genetic haplotypes were determined via the COI sequences. The ITS1 and the COI sequences revealed an important genetic variability within the Mediterranean population of C. glaucum, which contained two different phenotypes: orange foot and yellow foot. A genetic differentiation between the two phenotypes was revealed by the nucleotide diversity index (p) and is strengthened by the AMOVA analysis. This result leads us to suspect the presence of two different groups in the same population of C. glaucum. Nevertheless to be confirmed this hypothesis requires further studies using more locations and a larger number of sample sizes.

The aim of this study was to measure the mercury (Hg) contents of three marine fish and common seafood species (Diplodus annularis, Sarpa salpa and Sepia officinalis) at two sampling sites in the gulf of Gabes, i.e. Sidi Mansour (polluted site) and Kerkennah (control site). These species are frequently consumed by the population living at the Sfax coasts of Tunisia, particularly by the families of fisherman. Additionally, the hair mercury levels of 55 volunteers (28 women, 27 men) were analysed and the daily total mercury intake through the fish and seafood diet was estimated. The key findings were: (1) the mercury contents of the examined fish and seafood species frequently exceeded the regulatory guideline value of 0.5 mg/kg, (2) no site-specific differences in hair mercury contents were found, (3) fish and seafood consumption is probably the major contributor of mercury exposure in this population, (4) the daily mercury intake through frequent consumption of D. annularis exceeds the US EPA reference dose. Further studies are necessary to evaluate the health risks associated with such high mercury exposure levels in order to allow optimal counseling and therapy of the concerned population and to avoid future impairment of human health, particularly children’s health.