Salmonella Broiler Meat's Contamination in Tunisia: Prevalence, Serotypes, Antimicrobial Resistance and Molecular Characterization of Isolated Strains.

This study was conducted in north-eastern Tunisia to estimate the contamination prevalence of Salmonella in broilers' meat, to rank serotypes and to characterize the isolated multidrug-resistant (MDR) strains. A total number of 1288 meat samples were collected from 322 broiler batches; Salmonella isolates were identified by the alternative technique VIDAS Easy Salmonella. The susceptibility of Salmonella isolates was assessed against 21 antimicrobials using the disc diffusion method on Mueller-Hinton agar. Some antimicrobial resistance genes were identified using Polymerase Chain Reaction (PCR). The prevalence rates of Salmonella in the neck skin and the breast muscle contamination were estimated at 11.8% (38/322) and 0.9% (3/322), respectively. The prevalence rate of Salmonella in meat cutting parts contamination was estimated at 5.1% (33/644). Eight serotypes of Salmonella were identified, namely S. Enteritidis, S. Kentucky, S. Anatum, S. Infantis, S. Mbandaka, S. Zanzibar, S. Hadar and S. Agona. High rate of resistance was identified against amoxicillin (91.9%), nalidixic acid (83.8%), tetracycline (75.7%), streptomycin (73%), ciprofloxacin (70%), sulfamides (68.9%), cefalotin (68.9%), cefotaxim (67.6%) and cefoxitin (60.8%). The majority (90.5%; 67/74) of isolated strains was recognized as MDR. Nine MDR strains were identified as Extended-Spectrum ß-Lactamase (ESBL) producers. The blaCTX-M gene was identified by PCR in all the nine ESBL strains. TetA, tetB and dfrA1 genes were amplified in 3.6% (2/56), 1.8% (1/56) and 19.3% (5/26) of tetracycline and trimethoprim-resistant strains, respectively. The integrase gene (class 2) was identified in only 8.1% (6/74) of the Salmonella-isolated strains. Our findings highlight the emergence of MDR Salmonella isolates in Tunisia.

Metal transfer budgets in a Mediterranean marine environment subjected to natural and anthropogenic inputs: case of the Mejerda River Delta (Gulf of Tunis, northern Tunisia).

Deltaic sediments are important for biogeochemical metal cycling since they are hotspots for metal inputs. In addition, they are potential sites for diagenetic processes leading to either the burial of inorganic contaminants or their release. Diffusive fluxes of certain metals (Fe, Mn, Pb, Zn, Cu and Cd) in the sediments of the Mejerda River Delta (MRD) (Gulf of Tunis, Tunisia) were quantified by modeling the available concentration profiles in the pore water. The metals' burial and sedimentation fluxes were also calculated using both the asymptotic concentrations of available metal profiles and sediment trap results. These fluxes were assembled with the exchange fluxes at the sediment-water interface in order to develop complete metal transfer budgets. The results showed that budgets of Cu and Zn are almost neutral. The sediment appears to be a good trap for iron since its average burial flux at the three studied stations is about 332.6 g m-2 year-1. Organic matter degradation, carbonate dissolution, and oxyhydroxide reduction are the main mechanisms which accelerate the release of metals associated with the suspended particle matter once they reach the pore water in the seabed.

Distribution and assessment of heavy metal toxicity in sediment cores from Bizerte Lagoon, Tunisia.

To examine the state of pollution of Bizerte Lagoon which is exposed to intense anthropogenic pressure, two sediment cores were taken at two sites, one undergoes the dual effects of both marine waters arriving from the Mediterranean Sea through the Channel, and also of freshwater from the Tinja River; the other core is located at the center of the lagoon where water depth is maximal (12 m). Heavy metal concentrations in the two cores were assessed, with calculated enrichment factors and geo-accumulation indexes. Core sediments were also studied for chemical speciation and their monosulfide contents were measured. Results from enrichment factors and geo-accumulation indexes show an accumulation of Cd, Zn, Cr, and Pb, while chemical speciation revealed a risk only from Cd and Mn. Comparison of sequential extraction values with those of acid volatile sulfides revealed that non-toxic effects may be caused by any of the studied metals in the sediment.

Assessment of trace elements and fluoride originating from phosphogypsum in the sediment of Gulf of Gabes (southeastern Tunisia): what are the potential sources of accumulation and bioavailability?

To assess potential impacts of industrial activities on the pollution status of Gulf of Gabes, twenty sediment and water samples along with phytoplankton enumeration were achieved at different stations with specific features. Comparing trace element concentrations in sediment to applicable SQG standards, we were intrigued by an accumulation of Zn, Cr, Ni, and especially Cd, which exhibited relatively high content compared to these standards. Moreover, trace metal bioavailability was high in front of industrial discharge areas. The chemical speciation pointed out a high affinity of Pb, Zn, Cr, Mn, Ni, Co, and Fe for the residual fraction of the sediment. Bioavailability of trace elements was confirmed in surface sediment by the presence of a potential toxic fraction especially in front of industrial discharge areas. Toxicity assessment performed for the first time in the Gulf of Gabes through SEM and AVS models pointed to a high potential risk near both Ghannouch and Gabes Ports. Finally, the correlations between phytoplankton species and the labile fraction inferred potential phytoplankton bioaccumulation of Zn, Cu, and Cd both in the seawater and in the labile fraction.

Serotype Occurrence, Virulence Profiles, Antimicrobial Resistance and Molecular Characterization of Salmonella Isolated from Hospitalized Patients with Gastroenteritis in Great Tunisia between 2010 and 2020.

Non-typhoid Salmonella is one of the major causes of food-borne infections worldwide. The aim of the current study is to determine the serotype occurrence, virulence factors and antimicrobial resistance patterns of Salmonella isolated from hospitalized patients. The identification of Salmonella strains was performed according to REMIC, 2018. The susceptibility of Salmonella isolates was assessed against 20 antimicrobials using the disk diffusion method. Some virulence and antimicrobial resistance genes were identified using PCR. Among the 61 isolated Salmonella strains, seven serotypes were identified and all were positive for the virulence genes invA, mgtC and sirA. Critical resistance rates (>40%) were detected for tetracycline, nalidixic acid, amoxicillin and fluoroquinolones. However, resistances to ertapenem, ceftazidim, aztreonam and colistin were null. In addition, 33% of the isolated strains were multidrug-resistant (MDR). Moreover, 80% and 60% of S. Kentucky isolates were identified as fluoroquinolone-resistant and MDR strains, respectively. The qnrB gene was amplified in 63.2% of fluoroquinolone-resistant strains. The dfrA1 gene was identified in 20% (4/20) of the trimethoprim-sulfamethoxazole resistant strains and the integrase Class 2 gene was amplified in only 8.2% (5/61) of the isolates. Our findings highlight the emergence of MDR Salmonella isolates. A rationalization of antimicrobial use is urgently recommended in both human and veterinary medicine.

Early diagenetic behavior of trace metals along with estimation of their diffusive fluxes: Ecological risk assessment in pore water and sediment of Bizerte Lagoon, Tunisia.

Bizerte Lagoon is a vital Mediterranean ecosystem subjected to intense anthropogenic pressure. The potential ecological risk caused by certain metals (Zn, Cu, Cr, Cd, Ni, Pb and Mn) is assessed from the data carried out in the sediment and pore water at two sites along with identifying the effects of diagenetic processes on the vertical distribution of these metals and their resulting diffusive fluxes. Using various ecological indices our results reveal a high ecological risk to benthic organisms from metals chiefly Cd, Pb and Ni accumulated in the sediment at both sites. Metals derived from organic matter degradation (Cu and Cd) and/or reduction of Mn-Fe-oxyhydroxydes (Pb, Ni, Zn, Cr) due to early diagenetic processes in sediment. The resulting concentration gradients between pore water and overlying water induce diffusive fluxes of metals to the water column. The estimation of the potential ecological risk caused by dissolved metals in pore waters by application of the Interstitial Water Criteria Toxic Units index indicates a slight ecological risk by Cu that was not identified in sediment. The ecological risk posed by dissolved metals is evidenced from -6 cm depth, which reduces the possibility of contamination of benthic species living above the water-sediment interface by diffusive fluxes of these metals.

Spatial distribution, contamination assessment and potential ecological risk of some trace metals in the surface sediments of the Gulf of Tunis, North Tunisia.

To evaluate the trace metals contamination status in the Gulf of Tunis, forty one sediment samples were analyzed using different approaches. According to certain contamination and ecological risk indices (Contamination Factor, Geoaccumulation index and Ecological risk index), Hg has the highest contamination level while pollution by Ni, Pb, Cd and Cr was absent. The highest concentrations of trace metals were found in sediments collected from the offshore and coastal areas located opposite the main exchange points with the gulf particularly, the Mejerda and Meliane Rivers, the Khalij Channel, Ghar El Melh and El Malah lagoons, Tunis Lake and Sebkhat Ariana. However, further ecological indices (Potential ecological risk index, Toxic unit and Mean effect-range median quotient) and comparison with sediment quality guidelines suggest that in addition to Mercury, Cr, Pb and Ni concentrations are detrimental to biota in both the offshore and areas near to the exchange points with the gulf. Moreover, in these areas the results from sequential extraction and individual contamination factor calculation pointed to the mobility and bioavailability of Cr, Pb and Ni.

Prevalence, Risk Factors, Antimicrobial Resistance and Molecular Characterization of Salmonella in Northeast Tunisia Broiler Flocks.

This study was conducted in northeastern Tunisia to estimate both the prevalence and the risk factors of Salmonella in broiler flocks as well as to characterize the isolated multidrug-resistant (MDR) Salmonella strains. In the present study, a total number of 124 farms were sampled; Salmonella isolates were identified by the alternative technique VIDAS Easy Salmonella. The susceptibility of Salmonella isolates was assessed against 21 antimicrobials using the disk diffusion method on Mueller-Hinton agar using antimicrobial discs. Some antimicrobial resistance genes were identified using PCR. The prevalence rate of Salmonella infection, in the sampled farms, was estimated at 19.9% (64/322). Moreover, a total number of 13 different serotypes were identified. High rate of resistance was identified against nalidixic acid (82.85%), amoxicillin (81.25%), streptomycin (75%), and ciprofloxacin (75%). Alarming level of resistance to ertapenem (12.5%) was noticed. A total of 87.5% (56/64) of isolated strains were recognized as MDR. Three MDR strains were extended-spectrum ß-lactamases (ESBL)-producers and three MDR strains were cephalosporinase-producers. The blaCTX-M gene was amplified in all the three ESBL strains. The qnrB gene was not amplified in fluoroquinolones-resistant strains. The tetA and tetB genes were amplified in 5% (2/40) and 2.5% (1/40) of tetracycline-resistant strains, respectively. The dfrA1 gene was amplified in five of the 20 trimethoprim-resistant strains. The mcr-1, mcr-2, mcr-3, mcr-4, and mcr-5 genes were not amplified in any of the phenotypically colistin-resistant strains. In terms of integrase genes int1 and int2, only gene class 2 was amplified in 11% (7/64) of analyzed strains. Risk factors, such as the poor level of cleaning and disinfection, the lack of antimicrobial treatment at the start of the breeding, and a crawl space duration lower than 15 days, were associated with high Salmonella infection in birds. These data should be considered when preparing salmonellosis control programs in Tunisian broiler flocks.

Contamination of Fishery Products with Mercury, Cadmium, and Lead in Tunisia: Level's Estimation and Human Health Risk Assessment.

The levels of metallic trace elements were determined in fishery products sampled from Tunisian fishing ports. Mean concentrations were 0.20 ± 0.01, 0.05 ± 0.01, and 0.10 ± 0.01 mg/kg in fish flesh for Hg, Cd, and Pb, respectively. The mean concentrations of these elements were below regulatory thresholds; however, we also detected some specimens with higher levels. The mercury level showed a significant difference between fish categories, region, and year of sampling (p < 0.05). Samples from the center sites had the highest levels of Pb (0.17 ± 0.03 mg/kg; range 0.10; 0.22 mg/kg). Temporal analysis of Pb showed a significant difference between sampling year (p < 0.05). In addition, distribution among fish categories was statistically significant (p = 0.046). The rates of samples exceeding limits of Hg, Cd, and Pb were 5.3, 2.6, and 0.4%, respectively. Through the above results, the maximum weekly and monthly intakes for fish flesh consumption could not exceed the set limits. These data were important to inform consumers about fish content and the risk generated by some of these species.

Sulfide influence on metal behavior in a polluted southern Mediterranean lagoon: implications for management.

The degree of pyritization and degree of trace metal pyritization (DTMP) were investigated in sediments from Ghar El Melh Lagoon (northern Tunisia) in order to study metal deposition. A sediment core and 28 samples were thus taken in summer 2008, and metals and sulfate were analyzed in pore water/pyrite. Acid-volatile sulfide and metals were simultaneously extracted from these two fractions and the role of pyrite in the metal cycling studied. To examine pyrite presence and mineralogical form in sediments, X-ray diffraction of the washed and decarbonated sediment was performed along with scanning electron microscopy. Results showed that pyrite is present in fromboidal and euhedral forms. Thermodynamic calculation highlighted the formation of metallic sulfides and the co-precipitation of metals with iron sulfides. The DTMP increases with depth, indicating that these metals are either sequestered as sulfides or that they co-precipitate with pyrite into the deep sediment.

How useful are geochemical and mineralogical indicators in assessing trace metal contamination and bioavailability in a post-restoration Mediterranean lagoon?

Tunis's Lake North (LNT), located on the Mediterranean Sea, and Tunisia's most important lagoon due to its economic value and its strategic position within the city, has recently undergone a vast sanitation project. To study the lagoon's level of metal pollution, three sediment cores were taken and the sediments were analyzed for trace and major elements, acid volatile sulfides (AVS), total organic carbon (TOC), and minerals. Results showed that TOC concentrations (0.2-3.1%) decreased following the lagoon's restoration. In addition, in comparison to historical data, concentrations of Cu (16-69.5 µg g-1), Zn (60.6-191.4 µg g-1), and Pb (13-100.9 µg g-1) also decreased. Enrichment factor calculation with respect to the crust and local background showed that the sediment had long been contaminated by human pollution and especially by Pb, Zn, and Cu. The AVS to simultaneously extracted metal (SEM) ratio revealed values generally less than 1 indicating no sediment toxicity risks. Statistics revealed a detrital origin for certain metals and a diagenetic origin for FeS2 and carbonate minerals.

Trace element accumulation and elutriate toxicity in surface sediment in northern Tunisia (Tunis Gulf, southern Mediterranean).

Metal concentrations in sediments were investigated in the Gulf of Tunis, Tunisia, in relation to anthropic activities along the Mejerda River and Ghar El Melh Lagoon, with effluents discharged into the gulf. Distribution of grain size showed that the silty fraction is dominant with 53%, while sand and clay averages are 34 and 12% respectively. Zn concentration increased in the vicinity of the Mejerda River while Pb was at its highest levels at the outlet of Ghar El Mehl Lagoon. Sediment elutriate toxicity, as measured by oyster embryo bioassays, ranged from 10 to 45% abnormalities after 24h, but no relation was found between metal concentration and sediment toxicity. The AVS fraction that represents monosulfide concentrations in the sediment was higher in the central part of the gulf than in the coastal zone. The results reveal the influence of AVS, TOC and grain size on metal speciation and sediment toxicity.

Suspended particulate matter fluxes along with their associated metals, organic matter and carbonates in a coastal Mediterranean area affected by mining activities.

A study of suspended particulate matter (SPM) fluxes along with their associated metals, organic matter and carbonates, was conducted off the Mejerda River outlet in May 2011 and in March and July 2012 at depths of 10, 20 and 40 m using sediment traps. SPM fluxes are more significant near the Mejerda outlet, especially in winter, but dissipate further offshore. Normalization reveals that the Mejerda is a major source of Pb, Zn, Cd, Cu, Ni, and Co, all of which are the result of human activities. In contrast, Fe, Mn and N are of authigenic origin. The enrichment factor shows that Pb, Zn and especially Cd are the most highly polluting metals off the Mejerda outlet. This confirms the trend observed on the shores of the Mejerda prodelta and is consistent with the type of mining activities conducted in the Mejerda catchment.

Bioavailability and assessment of heavy metal pollution in sediment cores off the Mejerda River Delta (Gulf of Tunis): How useful is a multiproxy approach?

Three core samples were taken from zones offshore from the Mejerda River Delta (Tunisia) and analyzed for major and trace elements to assess their relationships with organic matter, monosulfides and carbonates, as well as for pollution and bioavailability. Chemical speciation, ∑ SEM/AVS, the enrichment factor (EF) and the geo-accumulation index (I-geo) were used. Iron, cadmium, lead and zinc - the most frequently mined metals in the Mejerda catchment - were found as contaminants in the offshore areas. Estimations of trace element accumulation using the EF and the I-geo index show that lead, and to a lesser extent zinc, are the most polluting metals off the Mejerda outlet. According to their bioavailability, these metals are also the most toxic. Only cadmium is heavily present in delta sediment (EF>100) though deeply sequestrated (100% bound to the residual fraction) and thus presents no toxicity.

Prevalence, virulence and antibiotic susceptibility of Salmonella spp. strains, isolated from beef in Greater Tunis (Tunisia).

The aim of this work was to investigate the presence of Salmonella spp. in 300 beef meat samples collected from cattle carcasses of different categories (young bulls, culled heifers and culled cows). The detection of Salmonella spp. was performed by the alternative VIDAS Easy Salmonella technique and confirmed by PCR using Salmonella specific primers. Salmonella serotypes were determined by slide agglutination tests. The resistance to 12 antibiotics was determined by the diffusion method on Mueller-Hinton agar antibiotic discs. The overall contamination rate of beef by Salmonella spp. was 5.7% (17/300). This rate varied from naught (0/100) in bulls' meat to 14% (14/100) in culled cows' meat (p<0.001). The prevalence of Salmonella spp. was higher in summer and in cattle with digestive disorders: chronic gastroenteritis (6/17), traumatic peritonitis (3/17) and intestinal obstruction (2/17) (p<0.0001). Of the 17 Salmonella isolates, 6 serotypes were identified, namely Salmonella Montevideo (8/17), Salmonella Anatum (3/17), Salmonella Minnesota (2/17), Salmonella Amsterdam (2/17), Salmonella Kentucky (1/17) and Salmonella Brandenburg (1/17) (p<0.05). Unlike other serotypes, S. Montevideo was present during the whole year except winter. Almost all of the strains (16/17) were resistant to at least one of the 12 tested antibiotics. Multidrug-resistance concerned 14/17 of the strains, including Amoxicillin (13/17), Tetracycline (12/17), Streptomycin (10/17) and Nalidixic acid (6/17). All the strains were sensitive to the association (Amoxicillin+Clavulanic acid), Cefoxitin and Ceftazidime. In addition, our study showed that all Salmonella strains (17) were positive for invasion gene invA and negative for the virulence gene spvC. Only one isolate (S. Kentucky) harbored the h-li virulence gene.

Determinants of urban sprawl in European cities.

This paper provides empirical evidence that helps to answer several key questions relating to the extent of urban sprawl in Europe. Building on the monocentric city model, this study uses existing data sources to derive a set of panel data for 282 European cities at three time points (1990, 2000 and 2006). Two indices of urban sprawl are calculated that, respectively, reflect changes in artificial area and the levels of urban fragmentation for each city. These are supplemented by a set of data on various economic and geographical variables that might explain the variation of the two indices. Using a Hausman-Taylor estimator and random regressors to control for the possible correlation between explanatory variables and unobservable city-level effects, we find that the fundamental conclusions of the standard monocentric model are valid in the European context for both indices. Although the variables generated by the monocentric model explain a large part of the variation of artificial area, their explanatory power for modelling the fragmentation index is relatively low.