First Investigation of Seasonal Concentration Behaviors and Sources Assessment of Aliphatic Hydrocarbon in Waters and Sediments from Wadi El Bey, Tunisia.

The contents, composition profiles, and sources of aliphatic hydrocarbons were examined in surface sediment and water samples collected from Wadi El Bey, in Tunisia, during different year seasons in 14 stations receiving domestic effluent, industrial discharge, and agricultural drainage wastes. The target substances were analyzed by gas chromatography coupled with mass spectrometric detection (GC/MS). Total concentrations of n-alkanes (n-C14-n-C38) ranged from 0.08 ± 0.01 to 18.14 ± 0.1 µg/L in waters and 0.22 ± 0.04 to 31.9 ± 24.6 µg/g in sediments, while total aliphatic fraction ranged from 0.08 ± 0.01 to 196 ± 140 µg/L in waters and 0.22 ± 0.04 to 1977 ± 1219 µg/g in sediments, which means that almost all sites were affected by hydrocarbon contents in sediments exceeding the recommended limit (100 µg/g). Various diagnostic indices (ADIs) were used to identify the hydrocarbon sources, namely the concentration ratios of individual compounds (n-C17/pristane, n-C18/phytane, pristane/phytane, n-C29/n-C17, n-C31/n-C19) as well as cumulative quantities (Carbon Preference Index, natural n-alkanes ratio, terrigenous/aquatic compounds ratio, unresolved complex mixture percentage, low molecular weight vs. high molecular weight homologues, Alkane Proxy and Terrestrial Marine Discriminants). In general, these indexes indicated that the origin of aliphatic hydrocarbons affecting sediments and waters of Wadi El Bey were linked to both biogenic and petrogenic inputs, attesting the impact of plankton and terrestrial plants and of oil contamination, respectively. The average carbon chain length computation (ACL), used to further index the chemical environment, ranged from 25.5 to 31.1 in sediments and 47.9-116 in waters. This finding could depend on the severe disturbances suffered by the ecosystem as a consequence of heavy anthropogenic inputs. Petroleum contamination associated with high eutrophication rates in Wadi El Bey must be strictly controlled, due to possible harmful effects induced on ecosystem and humans.

Seasonal occurrence, source evaluation and ecological risk assessment of polycyclic aromatic hydrocarbons in industrial and agricultural effluents discharged in Wadi El Bey (Tunisia).

Polycyclic aromatic hydrocarbons are of great concern due to their persistence, bioaccumulation and toxic properties. The occurrence, source and ecological risk assessment of 26 polycyclic aromatic hydrocarbons in industrial and agricultural effluents affecting the Wadi El Bey watershed were investigated by means of gas chromatographic/mass spectrometric analysis (GC/MS). Total PAHs (∑ 26 PAH) ranged from 1.21 to 91.7 µg/L. The 4- and 5-ring compounds were the principal PAHs detected in most of 5 sites examined. Diagnostic concentration ratios and molecular indices were performed to identify the PAH sources. Results show that PAHs could originate from petrogenic, pyrolytic and mixed sources. According to the ecotoxicological assessment, the potential risk associated with PAHs affecting agricultural and industrial effluents ranged from moderate to high for both aquatic ecosystem and human health. The toxic equivalency factor (TEF) approach indicated that benzo[a]pyrene and benz[a]anthracene were the principal responsible for carcinogenic power of samples.

Seasonal Distribution, Source Identification, and Toxicological Risk Assessment of Polycyclic Aromatic Hydrocarbons (PAHs) in Sediments from Wadi El Bey Watershed in Tunisia.

Surface sediments were collected from the Watershed of Wadi El Bey in Tunisia to evaluate the degree of polycyclic aromatic hydrocarbons (PAHs) contamination. Sediments were collected during different seasons in 14 sites that received wastes from domestic effluent, industrial discharge, and agricultural drainage wastes. Twenty-six individual PAHs were analyzed. The total PAH contents (Σ PAHs) in surface sediments showed wide variability, ranging from 6.89 ± 0.05 to 340 ± 0.1 ng g-1. The 4-, 5-, and 6-ring compounds were the most abundant PAHs detected at the majority of sites. Diagnostic concentration ratios between pairs of PAHs and molecular indices, calculated with the purpose of drawing information about pollution sources, indicated that PAHs were of both petrogenic and pyrolytic origins. Toxic contaminants concentrations were determined according to the numerical effect-based sediment quality guidelines (SQGs). PAH levels did not exceed the SQGs, indicating that PAHs seem to pose low and occasional toxicity risks. Total carcinogenicity and mutagenicity (TEQBaP and MEQBaP) ranged from 0.08 to 65 ng and from 0.02 to 135.0 ng g-1 of dry weight, respectively. Among the seven carcinogenic PAHs, BaP accounted for the majority of the potency and could potentially be used as a unique indicator of PAH toxicity. This study provides a baseline to promote environmental protection programs and pollution monitoring/control in Watershed and coastal areas.

Eukaryotic molecular diversity at different steps of the wastewater treatment plant process reveals more phylogenetic novel lineages.

Wastewater microbiota represents important actors of organic depollution. Nowadays, some species used as bioindicators of the effluent quality are still identified by microscopy. In the present study, we investigated eukaryotic diversity at the different steps of the treatment process of a wastewater treatment plant (aerobic, anaerobic, clarifier basins and anaerobic digester) using the 18S rRNA gene sequencing approach. Of the 1519 analysed sequences, we identified 160 operational taxonomic units. Interestingly, 56.9% of the phylotypes were assigned to novel phylogenetic molecular species since they show <97% sequence identity with their nearest affiliated representative within public databases. Peritrichia ciliates were the most predominant group, with Epistylis as the most common genus. Although anaerobic, the digester appears to harbor many unclassified phylotypes of protozoa species. Novel lineages such as LKM11 and LKM118 were widely represented in the digester. Diversity values given by Shannon indexes show that the clarifier is the most diversified. This work will help designing molecular tools that are fast, reliable, and reproducible for monitoring wastewater depollution and studying phylogenetic relationships among the wonderful world of protists within this anthropogenic ecosystem.

Combined effects of metal contamination and abiotic parameters on biomarker responses in clam Ruditapes decussatus gills: an integrated approach in biomonitoring of Tunis lagoon.

The spatial and seasonal alteration in a battery of biomarkers responses (enzymatic activity of glutathione-S-transferase, catalase and acetylcholinesterase and lipid peroxidation) were investigated to assess the metal derived effects in clam (Ruditapes decussatus) gills, collected from Tunis lagoon (Tunisia). Trace metals (Ag, As, Cd, Cu, Hg, Mn, Ni, Pb, V and Zn) concentrations were assessed seasonally in sediments and tissues of R. decussatus from three different sites (S1, S2 and S3). Bioaccumulation factor (BAF) analysis showed a spatio-temporal variation of metal uptake rates in clams through sediments. Likewise, the multibiomarker approach enabled a time-site trend differentiation between sites with distinctive degrees of anthropogenic contamination. Site S2 was identified as the most impacted region due to the presence of different contamination sources (shipping and industrial activities). The results suggest that biomarker's seasonal variation arises from a complex interaction between environmental conditions (e.g. temperature, salinity) and probably biological factors (mainly the reproduction process) along with anthropogenic pressure. The general biological response measured with the IBR index at all the sampling sites revealed the highest metabolic stress in summer. The combined effects of metal contamination and increased temperature and salinity in summer appear to induce the highest metabolic adaptation response. The selected biomarkers provided an integrated response, which is useful for the assessment of the combined effects of metal contamination and abiotic parameters in clams and the environmental status of coastal lagoon ecosystem.

Biological Monitoring of Occupational Exposure to Polycyclic Aromatic Hydrocarbons at an Electric Steel Foundry in Tunisia.

Occupational exposures during iron and steel founding have been classified as carcinogenic to humans, and the exposure to polycyclic aromatic hydrocarbons (PAHs) in this industrial setting may contribute to cancer risk. The occupational exposure to PAHs was assessed in 93 male workers at an electric steel foundry in Tunisia by biomonitoring, with the aims of characterizing the excretion profile and investigating the influence of job title and personal characteristics on the biomarkers. Sixteen 2-6 ring unmetabolized PAHs (U-PAHs) and eight hydroxylated PAH metabolites (OHPAHs) were analyzed by gas chromatography-triple quadrupole tandem mass spectrometry and liquid chromatography triple quadrupole tandem mass spectrometry, respectively. Among U-PAHs, urinary naphthalene (U-NAP) was the most abundant compound (median level: 643ng l(-1)), followed by phenanthrene (U-PHE, 18.5ng l(-1)). Urinary benzo[a]pyrene (U-BaP) level was <0.30ng l(-1) Among OHPAHs, 2-hydroxynaphthalene (2-OHNAP) was the most abundant metabolite (2.27 µg l(-1)). Median 1-hydroxypyrene (1-OHPYR) was 0.52 µg l(-1) Significant correlations among urinary biomarkers were observed, with Pearson's r ranging from 0.177 to 0.626. 1-OHPYR was correlated to benzo[a]pyrene, but not to five- and six-rings PAHs. A multiple linear regression model showed that job title was a significant determinant for almost all U-PAHs. In particular, employees in the steel smelter workshop had higher levels of high-boiling U-PAHs and lower levels of low-boiling U-PAHs than those of workers with other job titles. Among OHPAHs, this model was significant only for naphthols and 1-hydroxyphenanthrene (1-OHPHE). Smoking status was a significant predictor for almost all biomarkers. Among all analytes, U-PHE and 1-OHPHE were the less affected by tobacco smoke, and they were significantly correlated with both low- and high-molecular-weight compounds, and their levels were related to job titles, so they could be proposed as suitable biomarkers of PAH exposure at steel foundries. Based on 1-OHPYR levels, our findings show that occupational exposure of these workers was similar to that reported in recent studies of electric steel foundry workers. The multianalytic approach is useful in revealing different exposure levels among job titles.

Transcriptomic, Biochemical, and Histopathological Responses of the Clam Ruditapes decussatus from a Metal-Contaminated Tunis Lagoon.

This study was designed to investigate the molecular (transcriptional expression), biochemical (oxidative stress and neurotoxicity), and histopathological effects of metal contamination in the gill of clams (Ruditapes decussatus) sampled from the Tunis lagoon. The concentrations of five heavy metals (Cd, Pb, Hg, Cu, and Zn) in surface sediments and their accumulation in soft tissues of R. decussatus were evaluated in three sites (Z1, Z2, and Z3). A metal contamination state of Tunis lagoon sediments was noted with spatial variations with relatively high levels at Z2. Biomarker analyses showed an increase in glutathione S-transferase and catalase activities and lipid peroxidation levels and a decrease in acetylcholinesterase activity in the studied sites. Molecular investigation showed a significant overexpression of: cytochrome c oxidase subunit I, ribosomal RNA 16S, Cu/Zn superoxide dismutase, heat shock protein 70, and metallothioneins in the three sampling sites. Moreover, our data were correlated to severe and diverse histopathological alterations in the clam gills. The principal component analysis showed that the Z2 region is more affected by metal contamination than Z1 and Z3 regions. Current field results suggest the use of several combined biomarkers at different cell levels instead of individual ones in monitoring programs.

Bacterial structure and spatiotemporal distribution in a horizontal subsurface flow constructed wetland.

In this study, bacterial community structure in a horizontal subsurface flow constructed wetland (HSF-CW) planted with Phragmites australis was investigated using the 16S rRNA cloning-sequencing technique. Two layer depths were considered: the rhizosphere zone (RH) and the deep-layer zone (DL) in different sampling periods. Bacteria-specific primers 008F and 1492R were used to amplify the 16S rRNA genes and construct six clone libraries. A total of 1,284 individual sequences were used to assess the HSF-CW diversity. Phylogenetic analysis of RH and DL clone libraries shows that 41.57 and 42.17 % of the 16S rRNA sequences are affiliated with the Proteobacteria in the RH and the DL, respectively. The remaining major phylogenetic groups are Bacteroidetes, Planctomycetes, and Chloroflexi with 11.78, 9.36, and 7.6 %, respectively, in the RH and 11.38, 6.48, and 7.65 % in the DL, respectively. Minor divisions such as Verrucomicrobia, TM7, Nitrospira, and Gemmatimonadetes represented <6 % of the total sequences, while 14.2 % were unidentified Bacteria. Among the Proteobacteria, the Alphaproteobacteria subclass is represented in both locations, while the Deltaproteobacteria and Gammaproteobacteria subclasses were predominant in the RH and the DL, respectively. Results suggest that Archaea and Bacteria in the HSF-CW are the essential actors in the nitrogen cycle and that the established microbial community is efficient in nitrogen removal from wastewater.