Trace element levels in the muscles of three tern species (Aves: Laridae) from the western Arabian Gulf: environmental assessment and implications for conservation.

In the Arabian Gulf (called also Persian Gulf; hereafter 'the Gulf'), Jana and Karan Islands are recognized as one of the most Important Bird Areas in the region. Many migratory breeding seabirds, like the Greater Crested Tern Thalasseus bergii, White-cheeked Tern Sterna repressa and Bridled Tern Onychoprion anaethetus, depend on these islands during the breeding season. However, these aquatic wildlife species are suffering from intensified urban and industrial coastal development and various contamination events including wars and related oil spills. In this study, we used these three piscivorous top predator birds to analyse the levels of 19 trace elements (TEs; i.e. Al, As, Ba, Ca, Cd, Co, Cr, Cu, Fe, Hg, K, Mg, Mn, Na, Ni, Pb, Sr, V and Zn) in 15 muscular tissue samples from Jana and Karan Islands. PERMANOVA analysis showed no difference in contamination profile between sites nor between species probably due to their spatial and ecological proximity and therefore similar levels of exposure to TEs. Comparing these levels with existing literature, our results showed no particular concern for all elements, except for Al (maximum values recorded = 116.5 µg g-1 d.w.) and, in two samples, Ba (33.67 µg g-1 d.w.) and Pb (5.6 µg g-1 d.w.). The results can be considered as an initial step for supplementary evaluations with a larger number of samples and specified time intervals for the collection of specimens. This study provided baseline information on the pollution status of these two ecologically important sites which require a continuous biomonitoring programme.

DNA barcoding of marine fishes from Saudi Arabian waters of the Gulf.

We used the cytochrome oxidase subunit I (coI) gene DNA to barcode 117 endemic Gulf and cosmopolitan Indo-West Pacific fish species belonging to 54 families and 13 orders. Novel DNA barcodes were provided for 18 fish species (Trachinocephalus sp., Nematalosa sp., Herklotsichthys lossei, Upeneus doriae, Trachurus indicus, Apogonichthyoides taeniatus, Verulux cypselurus, Favonigobius sp., Suezichthus gracilis, Sillago sp., Brachirus orientalis, Pegusa sp., Lepidotrigla bispinosa, Lepidotrigla sp., Grammoplites suppositus, Hippichthys sp., Paramonacanthus sp. and Triacanthus sp.). The species delimitation analysis, conducted with Poisson tree processes- Bayesian PTP (PTP-bPTP) and nucleotide-divergence-threshold (NDT) models), found 137 and 119 entities respectively. Overall, NDT method, neighbour-joining species tree and the prior taxonomic assessment provided similar results. Among the 54 families considered, only 10 (Ariommatidae, Ephippidae, Leiognathidae, Nemipteridae, Plotosidae, Pomacanthidae, Pomacentridae, Priacanthidae and Rachycentridae) showed the occurrence of molecular diagnostic pure characters. The DNA barcoding database developed during this study will help ichthyologists to identify and resolve the taxonomic ambiguities they may encounter with the fishes occurring in The Gulf and throughout the region.

Molecular characterization of the full-length genome sequences of HDV strains circulating in Tunisia.

While Tunisia is endemic for hepatitis B virus (HBV), a recent large-scale retrospective study, revealed a very low prevalence (2%) of hepatitis Delta virus (HDV) (Yacoubi et al. in J Clin Virol 72:126-132, 2015). All strains were classified within the genotype 1 (HDV-1) as assessed by nucleotide sequencing of the so-called 'R0' region of the genome described previously. In this study, we aimed to determine the full-length genome sequence of HDV isolates in order to fully characterize the HDV strains spreading in Tunisia. Eleven HDV antibody and RNA positive samples were obtained from the 1615 clinical samples previously studied. The whole genome sequence was obtained for 5 strains by sequencing and realignment of four overlapping regions covering the entire genome, followed by extensive phylogenetic analyses. Tunisian sequences segregated together with Turkish and African sequences and showed 60% GC content. Alignment with an HDV-1 consensus sequence revealed that they exhibited several point mutations in different functional domains of the delta proteins that, according to previous studies, might possibly affect their properties. In conclusion, the first full-length genome sequences of Tunisian HDV isolates are provided, isolates which are closely related to Turkish and Sub-Saharan Africa strains, supporting the hypothesis for the spread of HDV-1-strains from Africa via Tunisia to Turkey, before spread to the rest of the world.

Economic losses related to the reduction of Posidonia ecosystem services in the Gulf of Gabes (Southern Mediterranean Sea).

In the early XXth century, the Gulf of Gabes in SE Tunisia used to host the most extended Posidonia oceanica seagrass beds in the Mediterranean basin and was a highly productive hotspot of benthic species. Since the 70's, >500 million t of wet toxic phosphogypsum discharges from a fertilizer industrial complex have led to the gradual loss of ∼90 % of its initial surface. This drastic shrinkage is accompanied by significant value losses originated from the direct and indirect-use services of which the most important ones are small scale fisheries and carbon storage function. Using market valuations of a number of services we estimate economic losses at 105 million € in 2014 (∼915€/ha), i.e., around 115 % of the added value of the gabesian fertilizer factories for the same year. Value losses should increase in the near future in relation with the COP26 agreements which boosted the open carbon credit market. Without actions to reduce negative production externalities caused by the fertilizer industry in the Gulf of Gabes it would not be possible to recover Posidonia ecosystems in this region leading to further economic, ecologic, and cultural losses.

PET plastics as a Trojan horse for radionuclides.

Mismanaged plastic waste interacts with secondary environmental pollutants, potentially aggravating their impact on ecosystems and human health. Here we characterized the natural and artificial radionuclides in polyethylene terephthalate (PET) bottles collected from the industrial littoral discharge of a phosphate fertilizer plant. The activity concentrations in littered bottles ranged from 0.47 (208Tl) to 12.70 Bq·kg-1 (226Ra), with a mean value of 5.30 Bq·kg-1. All the human health risk assessment indices (annual intake, annual effective dose, and excess lifetime cancer risk) estimated for radionuclides associated with ingestion and inhalation of microplastics were below international safety limits. Our results demonstrated that PET can be loaded with natural and artificial radionuclides, and potentially act as a carrier to transfer radionuclides to humans, posing a new potential health risk. Increased use, mismanagement and fragmentation of plastic waste, and continued interaction of plastic waste with radioelements may lead to enhanced radiation exposure in the future.

Bioaccumulation of trace elements and hydrocarbons in chondrichthyans of the western Arabian Gulf: Environmental and human health risk assessment and implications for conservation.

This is a first attempt to assess the levels of trace elements, PAHs, and TPHs in six elasmobranch species (Carcharhinus dussumieri, C. sorrah, Chiloscyllium arabicum, Gymnura poecilura, Sphyrna lewini, S. mokarran) from the Arabian Gulf. The chemical analysis showed that the concentrations of contaminants differed significantly between liver and muscle samples with both pelagic and benthic species and all families. For all species, contaminant concentrations were significantly higher in the liver than in the muscle. While muscle contaminant concentrations differed significantly with respect to species' lifestyles and families, those of liver showed no significant differences between pelagic and benthic species, neither between the four families nor between the six species. None of the analysed contaminants exceeded the internationally recognized standards in all studied species. These results enrich the knowledge on the bioaccumulation of contaminants in elasmobranchs and allow to assess the environmental status of the Arabian Gulf.

Seasonal patterns of microplastics in surface sediments of a Mediterranean lagoon heavily impacted by human activities (Bizerte lagoon, Northern Tunisia).

In this paper, we investigated seasonal variations in concentrations of microplastics (MPs) in surface sediments of a lagoon heavily impacted by human activities, located in northern Tunisia (the Bizerte lagoon, southern Mediterranean Sea). Analyses of 112 sediment samples collected from 28 stations between May 2019 and February 2020 revealed significant seasonal variation in concentrations of total MPs, with the highest levels recorded in August 2019 (109.6 ± 59.8 items kg-1 DS (dry sediment)) and the lowest in February 2020 (33.2 ± 22.0 items kg-1 DS). In terms of polymer types, polyethylene particles were the most abundant throughout the year, followed by polypropylene, polyvinyl chloride, and polyethylene terephthalate. Spatial variations in total MP concentrations were found to depend on several environmental factors, including proximity to the coastline, level of anthropogenic pressure, location inside the lagoon, and presence/absence of vegetation. The upper 5-cm surface sediment layer of the lagoon was found to contain ~ 9.96 × 1010 MP particles, equal to ~ 248.97 t of plastic. Similar patterns of microplastic composition and structure were found throughout the year, revealing the same plastic pollution hotspots during all seasons. This indicates that sources of plastic pollution are land-based and originate from coastal urban, industrial, and agricultural areas, as well as from major freshwater streams. The findings of the present work can help to develop an efficient environmental management plan aiming to reduce and/or stop the spread of plastic pollution and its impacts on the socially and economically important ecosystem of the Bizerte lagoon.

Whole genome analysis of hepatitis B virus before and during long-term therapy in chronic infected patients: Molecular characterization, impact on treatment and liver disease progression.

Hepatitis B virus (HBV) infection remains a serious public health concern worldwide despite the availability of an efficient vaccine and the major improvements in antiviral treatments. The aim of the present study is to analyze the mutational profile of the HBV whole genome in ETV non-responder chronic HBV patients, in order to investigate antiviral drug resistance, immune escape, and liver disease progression to Liver Cirrhosis (LC) or Hepatocellular Carcinoma (HCC). Blood samples were collected from five chronic hepatitis B patients. For each patient, two plasma samples were collected, before and during the treatment. Whole genome sequencing was performed using Sanger technology. Phylogenetic analysis comparing the studied sequences with reference ones was used for genotyping. The mutational profile was analyzed by comparison with the reference sequence M32138. Genotyping showed that the studied strains belong to subgenotypes D1, D7, and D8. The mutational analysis showed high genetic variability. In the RT region of the polymerase gene, 28 amino acid (aa) mutations were detected. The most significant mutations were the pattern rtL180M + rtS202G + rtM204V, which confer treatment resistance. In the S gene, 35 mutations were detected namely sP120T, sT126S, sG130R, sY134F, sS193L, sI195M, and sL216stop were previously described to lead to vaccine, immunotherapy, and/or diagnosis escape. In the C gene, 34 mutations were found. In particular, cG1764A, cC1766G/T, cT1768A, and cC1773T in the BCP; cG1896A and cG1899A in the precore region and cT12S, cE64D, cA80T, and cP130Q in the core region were associated with disease progression to LC and/or HCC. Other mutations were associated with viral replication increase including cT1753V, cG1764A/T, cC1766G/T, cT1768A, and cC1788G in the BCP as well as cG1896A and cG1899A in the precore region. In the X gene, 30 aa substitutions were detected, of which substitutions xT36D, xP46S, xA47T, xI88F, xA102V, xI127T, xK130M, xV131I, and xF132Y were previously described to lead to LC and/or HCC disease progression. In conclusion, our results show high genetic variability in the long-term treatment of chronic HBV patients causing several effects. This could contribute to guiding national efforts to optimize relevant HBV treatment management in order to achieve the global hepatitis elimination goal by 2030.

Surface sediment enrichment with trace metals in a heavily human-impacted lagoon (Bizerte Lagoon, Southern Mediterranean Sea): Spatial distribution, ecological risk assessment, and implications for environmental protection.

Although several studies previously assessed the contents of trace metals in the sediments of the heavily human-impacted lagoon of Bizerte (northern Tunisia), multi-analytical approaches have not been, so far, used to assess the ecological risks in this water body. This study attempts to provide a comprehensive ecological risk assessment related to the enrichment of the lagoon sediments with seven metals (As, Cd, Cr, Cu, Hg, Pb, and Zn). Significant spatial variations were found in the metal concentrations in sediments, in relation to the degree of coastal human activities and hydrodynamics. This was confirmed with the results of the three pollution indices, Cf, PLI, and Igeo. Concordant results were found with most of the indices used to assess the ecological risks (PERI, PEL, ERL, ERM, M-ERM-Q, TU), indicating higher risks in the southern part of the lagoon. These findings can help to improve the environmental management plan of the socio-economic important lagoon of Bizerte.

Rare earth elements characterization associated to the phosphate fertilizer plants of Gabes (Tunisia, Central Mediterranean Sea): Geochemical properties and behavior, related economic losses, and potential hazards.

This is the first study on the behavior and industrial fluxes of rare earth elements (REE) in the coastal fertilizer plants of Gabes (south-eastern Tunisia), the economic losses related to their wastes, and their environmental and human health hazards. The concentrations of 16 REE were assessed in phosphate rock (PR), phosphogypsum (PG) and phosphogypsum foam (PGF) samples, collected from Gabes plants. REE concentrations ranged from 0.23 (for Sc in PG) to 309.33 mg kg-1 (for Ce in PGF). Ce was the most abundant in the three matrices, with concentrations ranging between 80.40 (in PG) and 309.33 mg kg-1 (in PGF). PGF was the most enriched with REE (1075.32 mg kg-1). The annual flow of REE from the fertilizer factories to the marine environment may reach 1523.67 t. The economic losses related to the discharge of phosphogypsum REE in the Gulf of Gabes (GG) was estimated at ~58 million US$ y-1. The potential hazards of discharged REE on the local environment and human health were also evaluated and discussed. These findings show the need for the development of a new industry exploiting REE from phosphogypsum wastes (short term) and phosphate ores (long term) which should lead to reduce its high environmental and human health footprint and to potential economic gains.

Primary structure and anticoagulant activity of fucoidan from the sea cucumber Holothuria polii.

Sea cucumber fucoidan is recently endowed with a variety of biological activities. In the present study, we studied the structure and anticoagulant effect of fucoidan from the sea cucumber Holothuria polii (Fuc-Hp). The Fuc-Hp was purified by anion exchange chromatography and its structure was characterized by FT-IR and NMR spectroscopy. Molecular weight measurements were performed by HPSEC-MALLS-dRI. Fuc-Hp anticoagulant activity was measured by activated partial thromboplastin and thrombin times, and by in vitro thrombin inhibition in the presence of antithrombin and heparin cofactor II. The assessment of thrombin generation was investigated using calibrated automated thrombography. Fuc-Hp with a high sulfate content (34.6%) and an average molecular mass of 1376.3 kDa was isolated from H. polii in amount of ~2.7 mg/g dry body wall. Primary structural analysis indicated that Fuc-Hp was mainly composed of a tetrafucose repeating unit branched by glucuronic acid. Fuc-Hp exhibited a high anticoagulant effect mediated essentially by heparin cofactor II and to lesser extent by antithrombin with IC50 values of 0.16 µg/mL and 0.5 µg/mL, respectively. Overall results showed a high anticoagulant activity of Fuc-Hp, which was attributed to the high sulfate content and abundance of disulfated fucose residues of H. polii fucoidan.

Development and characterization of bioactive edible films from spider crab (Maja crispata) chitosan incorporated with Spirulina extract.

Active food packaging films based on crab chitosan and Spirulina extract (SE) were developed. The effects of the SE incorporation at different levels on physical (color, opacity water vapor and oxygen permeability) and mechanical (tensile strength and elongation at break) properties of chitosan films were investigated. FTIR was carried out to observe the potential modifications of the chitosan films when incorporated with SE. The obtained results suggested that incorporation of SE into chitosan films improved mechanical and barrier properties. The antioxidant activity of the chitosan/SE films was characterized by means of three different analytical assays (DPPH, FRAP and FIC). Crab chitosan edible films containing SE showed higher antioxidant activity, regardless concentrations and methods assayed. Furthermore, the antioxidant activity occurred in a concentration-dependent manner. The agar disc diffusion method was used to determine the antibacterial activities of chitosan edible films against Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Listeria monocytogenes, Salmonella typhimurium, Bacillus subtilis and Bacillus cereus. The chitosan/SE films were more effective (p<0.05) against five of the seven tested bacteria. The obtained crab chitosan edible films incorporated with SE showed great potential to be used for active food packaging due to its excellent antioxidant and antibacterial activities.

Molecular epidemiology of hepatitis B and Delta virus strains that spread in the Mediterranean North East Coast of Tunisia.

BACKGROUND: Tunisia is classified as an area of middle endemic for hepatitis B virus (HBV) infection, however little is known about hepatitis Delta virus (HDV) infection. OBJECTIVES: This study aimed to address the prevalence of HDV infection, to identify possible risks factors, and to analyze the genetic diversity of HDV strains that are spreading in Tunisia. STUDY DESIGN: A retrospective large-scale study including 1615 HBsAg positive patients, native of the North East coast of Tunisia, recruited from Gastroenterology departments, was conducted. Demographic, epidemiological, ethnical, clinical and biological data were recorded. HBV and HDV serological analyses and DNA and RNA viral load quantification were performed. Genotyping of HBV and HDV strains was performed using nucleotide sequencing followed by phylogenetic analyses. RESULTS: The study population included 819 (50.7%) men and 796 (49.3%) women; aged 12-90 years (mean age 41±13 years). A very low prevalence of HDV infection, 2% was observed. No risk factor, except a history of hospitalization for surgery was found. All HDV strains belonged to genotype 1, with a wide distribution within the HDV-1 group. They all share the African amino acid marker, a serine at position 202 of the large Delta protein. HBV genotypes were distributed as follows: HBV/D1 (56.8%), HBV/D7 (40.9%), and HBV/A2 (2.3%). CONCLUSION: Tunisia is a low endemic region for HDV infection, due to an efficient policy of HBV infection control. HDV-1 is the sole genotype found, with a high diversity within this group. Further studies are ongoing in order to better characterize and manage the HBV/HDV-infected patients according to the genetic variability of the viral strains.