Physicochemical and ecotoxicological approaches for Moknine Continental Sebkha in Tunisia.

Degradation of water quality is an emerging issue in many developing countries. In this context, industrial and domestic effluents heavily contaminate the coast of Moknine Continental Sebkha in Tunisia. The present study aimed to biomonitor the seawater quality of the Moknine Continental Sebkha coast using physicochemical and ecotoxicological approaches. The ecotoxicological assessment was performed using three species representing different trophic levels, namely Vibrio fischeri, Selenastrum capricornutum, and Lepidium sativum. In the physicochemical analysis such as BOD (biochemical oxygen demand), COD (chemical oxygen demand), TSS (total suspended solids), TOC (total organic carbon), NO3- (nitrate), AOX (adsorbable organic halogen), the recorded levels of pH and total suspended solids did not comply with the Tunisian standard (NT.09.11/1983). The ecotoxicological data confirmed that the tested water samples displayed toxicity to two test indicators L. sativum and S. capricornutum. A targeted chemical screening of the Moknine Continental Sebkha coast previously performed revealed the presence of total mercury, four phthalate acid esters, and one non-phthalate plasticizer, a fact that could explain the observed ecotoxicological effects and therefore might harm the biotic area and the health of the surrounding population.

Endocrine-disruptor endpoints in the ovary and thyroid of adult female rats exposed to realistic doses of di-(2-ethylhexyl) phthalate.

Di-(2-ethylhexyl) phthalate (DEHP) is the world's most widely used polyvinyl chloride (PVC) plasticizer and is used in virtually every category of flexible PVC. In fact, DEHP is extensively used in food cosmetics and medical packaging. It has become a serious problem in recent years. DEHP can be absorbed into the human body through the air, food, water, and skin. The current study involved intraperitoneal injection of DEHP dissolved in corn oil once daily for 21 consecutive days to investigate the effects of DEHP on the thyroid and the reproductive system in female rats. Results show that ovarian hormones (progesterone and estrogen) decreased significantly in the rats treated with DEHP compared to control. This result is supported by the alteration of folliculogenesis, the decrease of the follicles viability, and the apoptosis of the granulosa cells observed on histological sections of ovary and thyroid in female rats exposed to low doses of DEHP. Histopathological study revealed that DEHP could damage thyroid tissue and disrupt these functions. We also observed cellular damage, particularly in the liver cells, and a significant increase in biochemical parameters such as alanine aminotransferase (ALT) and aspartate aminotransferase (AST) compared to the control group.

Estrogenic hazards of short chain phthalates and bisphenols found in cosmetic products.

For several years, environmental exogenous agents, called endocrine disruptors, are suspected to interfere with the essential functions of reproduction and development in many living organisms. In this study, endocrine disruptors including five phthalates and two bisphenols contents in finished products were assayed and their estrogenic activity were measured by using the Yeast Estrogen Screen system with respect to human and trout estrogen receptors hERα and rtERS. Independently of the estrogen receptor, only short-chain phthalates (DBP and BBP) and the two bisphenols exhibited an estrogenic activity. Besides, the risk of three end-products (agro-food, cosmetics, and pharmaceutical) was evaluated before and after forced aging. Only two cosmetics the face cream and the perfume presented a hazard which increases with aging. These results are consistent with the compounds identified by Gas chromatography-mass spectrometry. These findings confirmed that the YES system can be routinely used to evaluate the estrogenic hazards within finished products.

Monitoring hospital wastewaters for their probable genotoxicity.

Hospitals' effluents contain a considerable amount of chemicals. Considering the significant volume of wastewater discharged by hospitals, the presence of these chemicals represents a real threat to the environment and human health. Thus, the aim of this study was to evaluate the in vivo and in vitro genotoxicities of three wastewater effluents collected from Tunisian hospitals. The liver of Swiss albino male mice, previously treated with different doses of the hospital wastewaters, was used as a model to detect DNA fragmentation. Our results showed all the hospital effluents caused significant qualitative and quantitative hazards in hepatic DNA. The wastewater collected from Sfax hospital exhibited the highest genotoxic effect, which may be explained by the presence in this effluent of some toxic micropolluants. There was a significant increase in genotoxicity, proportionally to the concentration of effluent. However, the vitotox assay did not show any significant genotoxicity on Salmonella typhimurium TA104 in the presence or absence of microsomal fraction S9. The ratio gentox/cytox was lower than the threshold 1.5. This study assessed the toxicological risk issued from Tunisian hospital wastewaters, which is potentially very harmful, and it has been pointed out that wastewater treatment requires special attention.

Human urine contamination with environmental pollutants: simultaneous determination using UPLC-MS/MS.

Paraben derivatives are widely used as an antifungal, antimicrobial preservative in cosmetic products, pharmaceuticals, and food. These molecules are called endocrine disruptors (EDCs). The exposure of the human body to paraben derivatives needs further study and for this purpose 200 urine samples were collected from Tunisian men and women aged between 5 and 90 years to determine three paraben derivatives: methylparaben (MP), ethylparaben (EP) and propylparaben (PP) using ultra performance liquid chromatography-tandem mass spectrometer (UPLC-MS/MS). The three major parabens were found in 95 urine samples. The obtained results indicate that MP, EP, and PP were detected in 57%, 46%, and 40% of all samples, respectively. Urinary concentration for the three parabens was in the range of 0.88-84.46 ng/mL, 0.52-29.2 ng/mL, and 0.51-28.17 ng/mL of PP, MP, and EP, respectively. In addition, the concentrations of the paraben derivatives in women were higher than those of men. These findings indicate that the exposure occurs from common products (foods, cosmetics, and pharmaceuticals). The Tunisian authorities should control the composition of packaging of these common products in order to protect humans against EDCs.

Organic contamination of Italian and Tunisian culinary herbs and spices.

Contaminants belonging to various classes, as polychlorobiphenyls, polycyclic aromatic hydrocarbons, organochlorine pesticides, organophosphorous pesticides, pyrethroid insecticides, fungicides, herbicides, synergists, carbamates, acaricides and insect growth regulators were simultaneously analysed by GC-MS/MS in 118 Italian and Tunisian culinary herbs and spices. The results obtained in Italian samples shown that laurel samples were the most contaminated with the presence of 15 residues on 140, whereas in rosemary (max value of 35 ng/g for cis-chlorfenyvinphos) and oregano (max value of 118.16 ng/g for ethion) some occasional residues can be observed, but always lower than the maximum residue levels; all the others samples shown no contamination. Among Tunisian samples, only rosemary contains a notably high content of pollutants exceeded the EU maximum residue limits (i.e., alachlor and phosalone with level of 359.2 and 43.3 ng/g, respectively), while oregano was determined to be free of contaminant residues. Considering the comparison among the different organic pollutants in Italian and Tunisian spices and herbs evaluated and the differences observed in this study a harmonization of regulation on contaminant residues in herbs and spices for human consumption should be needed, considering their increased use in diet and cooking.

Preliminary evaluation of plasticizer and BPA in Tunisian cosmetics and investigation of hazards on human skin cells.

In this study, 18 plasticizer (phthalates, adipates, sebacates, and others) and BPA residues in some cosmetic samples collected from Tunisian market are evaluated by micro-matrix solid-phase dispersion combined with GC-MS. In parallel, the impact of these molecules and the cosmetics in the human epithelial cell lines is investigated. The cytotoxic activity of cosmetic extracts is evaluated in vitro against B16 and Hep-2 human skin cell lines using the 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide assay. This study shows that the tested cosmetic products could constitute a hazard to the consumer health and wellness and that strict safety analysis on cosmetic products needs to be carried out before they are marketed.

Designation of pathogenic resistant bacteria in the Sparusaurata sea collected in Tunisia coastlines: Correlation with high performance liquid chromatography-tandem mass spectrometry analysis of antibiotics.

Vibrio is characterized by a large number of species and some of them are human pathogens causing gastro intestinal and wound infections through the ingestion or manipulation of contaminated fishes including Vibrio parahaemolyticus and Vibrio alginolyticus. In this study, we reported the phenotypic and molecular characterization of Vibrio parahaemolyticus and Vibrio alginolyticus strains isolated from wild and farm sea bream (Sparus aurata L.) along the Tunisian coast from December 2015 to April 2016. Therefore, the antibiograms indicate a difference between farmed and wild fish. Resistance against amoxicillin antibiotic appears for the bacteria isolated from wild fish, while those from aquaculture farming presented sensitivity to amoxicillin and resistance to antibiotics colistin and fusidic acid. The chloramphenicol antibiotic exhibited a high sensitivity in all isolated bacteria. In fact, traces of amoxicillin in the organs of the fish from Hergla farm were detected by UPLC-MS/MS analysis during December 2016 to April 2016. In addition, antibiotics were detected in January 2014 with high concentration of norfloxacin 2262 ng/g in fish from Hergla coast. The results obtained in this work indicated that the use and presence of antibiotics in water impacts on the occurrence of resistant bacteria and the detection of antibiotic in fish.

Occurrence and seasonal variation of plasticizers in sediments and biota from the coast of Mahdia, Tunisia.

Plasticizers are compounds often involved in the manufacturing of plastic products. Nevertheless, the ageing of the latter generates plasticizers that generally end up in the marine environment. In fact, marine pollution by phthalate acid esters (PAEs) and their alternatives has become an environmental and health issue of serious concern, as they are largely and ubiquitously present in the environment and aquatic organisms. In the present study, four PAEs, such as diethyl phthalate (DEP), diisobutyl phthalate (DiBP), dibutyl phthalate (DBP), and di(2-ethylhexyl) phthalate (DEHP), and one non-phthalate plasticizer (NPP), namely di-2-ethylhexyl terephthalate (DEHT), are wanted in different marine compartments from the coast of Mahdia in Tunisia such as sediment, seagrass, and mussel. The most abundant and frequently detected congener was DEHT at the concentrations reached 1.181 mg/kg in the sediment, 1.121 mg/kg in the seagrass, and 1.86 mg/kg in the mussel. This result indicates that the DEHT could emerge through the food chain and therefore bioaccumulate in marine compartments. In addition, we noticed that the seasonal variations of plasticizers were seriously affected by environmental factors including industrial and urban discharges.

Coastal Surveillance and Water Quality monitoring in the Rejiche Sea-Tunisia.

The aim of this study is to determine physiochemical and bacteriological properties of seawater and sediments from the coast of Tunisia during six successive months. The conductivity was measured using previously calibrated Hach Conductivity meter. Total suspended solid was determined using Whatman GF/C glass fiber filter (Sigma Aldrich), and the turbidity was assessed using a spectrophotometer (UV/Vis). The pH was measured using pH electrodes. Other physiochemical parameters were determined using Pastel UV multiparameter water analyzer (Secomam, aqualabo). Bacterial analysis was displayed by membrane filtration method, and isolates were identified to the species level by Api strips. Susceptibility to antibiotics was determined by disk diffusion methods. Total suspended solid, turbidity, adsorbable organic halogen, chemical oxygen demand, and biochemical oxygen demand were higher than accepted norms in Tunisia. Enterococcus faecalis was detected in all the sites indicating a human fecal contamination, and all the isolates were highly resistant to rifampicin, gentamicin, chloramphenicol, ampicillin, and azithromycin. The situation in Mahdia coast is alarming particularly with the isolation of multidrug resistance strains. It is recommended that the local government provide restrict policies for the treatment and the assessment of municipal wastewater before its discharge into sea. PRACTITIONER POINTS: Coastal Surveillance and Water Quality monitoring in the Rejiche Sea - Tunisia is crucial. Enterococcus faecalis was detected in sea water and sediment samples in the coast of Rejiche. Isolated strains were highly resistant to rifampicin, gentamicin, chloramphenicol, ampicillin and azithromycin.

Identification and quantification of plasticizers, bisphenol, and environmental toxic mineral elements residues in medicines from Tunisian markets.

In this study, 18 plasticizer (phthalates, adipates, sebacates, and others) residues in sixteen medicines available in Tunisian pharmaceutical markets are evaluated by MSPD combined with GC-MS. In parallel, in this research, UPLC-MS/MS technique was performed to detect bisphenol A, and cadmium, lead, chromium, cobalt, nickel, mercury, selenium, and arsenic levels were determined by ICP-MS. The maximum daily ingested mass of mineral elements was calculated and compared with permissible daily exposure limits published by USP NF 2013. Our results demonstrated that the 50% of drugs contained similar concentrations of di-ethyl adipate. Di-isobutyl phthalate was found in a single sample with a high concentration (1.07mgkg-1). Di-methyladipate was also present in only one sample. Diethyl phthalate was detected in three medicines: the highest concentration (17.03mgkg-1) was found in capsules. The other plasticizers and bisphenol A levels were below the limits of quantification in the all samples. All tested products were safe for use, except two clay-originated products. Concentrations and the maximum daily ingested mass of lead and arsenic were higher than recommended limits. The use of these medicines should require more attention taking into account the serious hazards of heavy metals to human health.

Evaluating the effect of dermaseptin S4 and its derivatives on multidrug-resistant bacterial strains and on the colon cancer cell line SW620.

Dermaseptins are peptides found in the skin secretions of Phyllomedusinae frogs. These peptides exert a lytic action on various microorganisms and have no considerable hemolytic effect except dermaseptin S4 (DS4) which exhibits a powerful cytotoxic effect. Therefore, we synthesized several analogs of DS4 in an attempt to find molecules with a weak hemolytic effect and significant bioactivities. In this study, we performed the synthesis of truncated peptides by introducing C-terminal and N-terminal amino acid deletions of the native sequence. All peptide analogs, in comparison with parental peptide, were tested firstly on human red blood cells to work out their cytotoxicity, secondly on the multidrug-resistant bacteria by trying to find MICs, and finally on colon cancer tumor cell line SW620 using the MTT test so as to investigate the anti-proliferative effect. Our results showed that, on the one hand, the N terminus of the native peptide was necessary for the antibacterial activity and the anti-proliferative effect of the peptide. On the other hand, the hemolytic activity was more notable in the sequences broken down on the C-terminal side.

Discrimination of Tunisian Honey by Mineral and Trace Element Chemometrics Profiling.

The concentrations of 19 chemical elements have been determined in 36 honey samples of different botanical (wildflower, eucalyptus, eucalyptus red flowers, prickly pears, lemon blossom, thyme, almond, rosemary and jujube) honeys from the three geographical areas of Tunisia (Sidi Bouzid, Nabeul and Sfax) using inductively coupled plasma mass spectrometry (ICP-MS). The aim of this work was to use the multielement analysis together with chemometric tools to verify the botanical and the geographical origin of honeys. The correlation on the basis of mineral element content between the honey samples and their botanical and/or geographical origins was in some measure achieved. The data collected on the samples were also used to evaluate the nutritional quality and the potential health risks associated with elements via consumption of the Tunisian honey. According to the results obtained, the intake of essential elements was small, and the potential health risks associated with toxic or potentially toxic elements via consumption of this food were overall insignificant.

Optimization of extraction with salicylic acid, rheological behavior and antiproliferative activity of pectin from Citrus sinensis peels.

A Box-Behnken design was used to optimize extraction temperature, extraction time and concentration of the salicylic acid to obtain a maximum polysaccharide yield from Citrus sinensis peels. The optimal settings were: extraction time 3 h, extraction temperature 80 °C and concentration of the salicylic acid 1.5%. Under these conditions, the experimental yield and uronic acid content were 11.74% and 66.9% respectively. Preliminary characterization was performed via FT-IR, SEC/MALS/VD/DRI and GC-MS after hydrolysis. SEC analysis showed that the extracted polysaccharide had a weight average molar mass of 350 kDa and an intrinsic viscosity of 640 mL/g. The GC-MS results revealed that the extracted polysaccharide was composed of arabinose 56.7%, galactose 17.8%, xylose 13.8%, rhamnose 5.1%, mannose 2.5% and glucose 1.5% suggested a rhamnogalacturonan pectin type I with a degree of esterification of 50.9% (IRTF). The flow curve and the dynamic frequency sweep were obtained at 10, 20, 30 and 40 g/L in water and at 30 g/L in presence of CaCl2 or NaCl at 1 mol/L. The solutions showed shear-thinning behavior fitted with Ostwald-De Waele model, except 10 g/L with a Newtonian behavior. The apparent viscosity and, the G' and G" moduli increase with PACO concentration in agreement with a slow-down of the dynamic chain. In the presence of CaCl2 or NaCl the reduction of electrostatic repulsions between pectin chains decreases the rheological parameters. The effect is less sensitive with CaCl2 due to intermolecular interactions. The antiproliferative activity of the extracted pectin on human Caco-2 and Hep-2 cells was very interesting with an IC50 1.4 and 1.8 µg/mL respectively.

Major, minor and trace element concentrations in spices and aromatic herbs from Sicily (Italy) and Mahdia (Tunisia) by ICP-MS and multivariate analysis.

Element contents in different types of spices and aromatic herbs collected from Italy and Tunisia were evaluated. The aim was to evaluate: the possible differences and/or similarities among the analyzed samples; if it is possible related the samples to their geographical origins; the nutritional quality and the potential health risks. Potassium, Ca, Mg and Ni were low in laurel and rosemary; mint and thyme showed the highest Na and the lowest Se contents; arsenic and Cd levels were found highest in verbena which had also the lowest Hg content; lastly, black pepper had the highest Mn and the lowest Pb contents. Instead discrimination between Italian and Tunisian samples for each spices and aromatic herbs under analysis was achieved by PCA. Essential elements intake through samples was small. Levels of Pb in some samples exceed the maximum allowable level, but any safety risk for consumers is excluded.

Organic pollution in PGI and non-PGI lemons and related soils from Italy and Turkey.

In this work 67 organic contaminants in Protected Geographical Indication (PGI) and non-PGI Interdonato Lemon pulps, peels, leave and related soils were evaluated by GC/MS-MS. It is possible to highlight that in Italian pulps and peels the residues were under LODs (0.10-10.30 ng·g-1), whereas the leaves showed piperonyl butoxide in 33% of samples (11.76 ± 1.24 ng·g-1) and cypermethrins in 33% of samples (Cypermethrin I, 6.12 ± 0.35 ng·g-1; Cypermethrin II, 8.67 ± 0.48 ng·g-1; Cypermethrin III, 8.90 ± 1.52 ng·g-1; Cypermethrin IV, 12.25 ± 2.46 ng·g-1). Turkish samples were mainly contaminated by piperonyl butoxide: it was revealed in the 33% of pulps (3.35 ± 1.03 ng·g-1), in 100% of peels (23.46 ± 17.69 ng·g-1) and in 100% of leaves (1302.44 ± 527.40 ng·g-1). In both soils a different contamination was detected. This study confirms that the European brand (PGI) of Interdonato Lemon Messina is a valuable instrument for the consumer protection.

Persistent plasticizers and bisphenol in the cheese of Tunisian markets induced biochemical and histopathological alterations in male BALB/c mice.

Clinical evidences, experimental models, and epidemiology of many studies suggest that phthalate-based plasticizers, aliphatic ester, and bisphenol A (BPA) have major risks for humans by targeting different organs and body systems. The current study has been designed firstly to analyze three categories of cheese with and without their exposure to the sun and packed in packages with an inner surface plastic-covered film in order to identify the dibutyl phthalate (DBP); benzyl butyl phthalate (BBP); bis(2-ethylhexyl) phthalate (DEHP); diisodecyl phthalate (DiDP); diisononyl phthalate (DiNP); and 1,2-cyclohexane dicarboxylic acid diisononyl ester (DINCH) by GC-MS/MS, except for the bisphenol A, which is by UPLC-MS/MS, and secondly to assess the toxicity of the identified chemical molecules and cheese samples on the liver and kidney of mice. Our results showed that the cheese contains high quantities of DBP and DEHP with the concentrations up to 0.46 and 2.339 mg/kg, respectively. Other types of cheese, such as rolled and triangular cheeses, contain little quantities of the all substances at concentrations below the standard limits. In vivo, the obtained data clearly demonstrated that the acute administration of DBP, DEHP, and the tested cheese significantly induced liver and kidney injuries in mice manifested by a rise in plasma alanine aminotransferase, aspartate aminotransferase, blood urea nitrogen, lactate dehydrogenase, urea, creatinine, and uric acid when compared with control animals. In addition, the histopathological study confirmed the perturbation of biochemical parameters and showed that the hepatic and renal structures were altered. Indeed, the hepatotoxic and nephrotoxic effects are more pronounced when cheese was exposed to the sun.

Plasticizers and BPA Residues in Tunisian and Italian Culinary Herbs and Spices.

In the present study, 18 plasticizers and residues in 10 different Tunisian and Italian culinary herbs and spices (black pepper, mint, caraway, coriander, oregano, rosemary, thyme, fennel, verbena, and laurel) were determined by GC/MS. Di-methylphthalate, di-(2-methylpropyl)adipate, di-n-butyladipate, di-propylphthalate, benzylbenzoate, di-phenylphthalate, and bisphenol A were lower than their LOQ in all 118 samples. Among the Tunisian samples, di-(2-ethylhexyl)phthalate was found in all types of samples and di-butylphthalate in 50% of types; all other phthalates were rarely dectected. Among the adipates, only di-methyladipate was found in 90% of types; di-ethyladipate was seldom found and di-(2-ethylhexyl)adipate only in samples of caraway. Di-(2-ethylhexyl)terephthalate was found in all types of samples; di-(2-ethylhexyl)sebacate was detected rarely but with high values. Among the Sicilian samples, di-ethylphthalate and di-(2-methylpropyl)phthalate were detected only in samples of mint; di-methyladipate, di-butylphthalate, and benzylbutylphthalate were identified in oregano and laurel; di-(2-ethylhexyl)terephthalate and di-(2-ethylhexyl)sebacate only in oregano. In any case, the results suggested that intake of these contaminants through spices and herbs is not a dangerous risk to the consumers. Probably, as already observed for the other food, these contaminants could result from pollution of the environment (air, water, and/or soil) and/or farming methods. PRACTICAL APPLICATION: Plasticizers and BPA in Tunisian and Italian spices were determined by GC/MS. Tunisian spices seem to contain more residues than Italian samples. Intake of these contaminants by spices is not a dangerous risk to the consumers. These pollutants could result from environmental pollution or agricultural practices.

In vivo protective role against water contamination with cerium via chronic administration of omega 3.

In the present study, adult, healthy male Wistar rats (120 ± 10 g) were pre-treated by intragastric administration of cerium chloride (CeCl3) 10 mg/kg (BW) each day during 60 days. Control animal were treated with omega 3, a polyunsaturated fatty acid (ω-3), by an intragastric administration at 10 mg/kg of BW for 60 days. Our results showed that CeCl3-induced alterations in all tested oxidative stress markers. In fact, CeCl3-induced the increase the level of both the creatinine concentration and the expression of lactate dehydrogenase, alkaline phosphatase, and transaminase activities in serum. On the other hand, CeCl3 significantly increased the levels of lipid peroxidation in the renal and hepatic tissues. The capacity of CeCl3 to generate reactive oxygen species (ROS) could explain his ability to induce morphological alterations, such as centrilobular hemorrhage, hepatic necrosis, and vacuolization of the cytoplasm in hepatic tissues, and the atrophy of the glomerulus and dilatation of urinary space in renal tissues. However, omega 3, after gastric administration, reduced significantly the toxic effect caused by CeCl3 according to his high ability to scavenge ROS. The present study indicates that omega 3 is a significant compound with protective activity against intoxication with heavy metal, the cerium, and thus may be useful for chemoprevention.

Cytotoxic effects of seven Tunisian hospital wastewaters on the proliferation of human breast cancer cell line MDA-231: correlation with their chemical characterization.

Hospital wastewaters contain large amounts of pharmaceutical residues, which may eventually be discharged into the aquatic environment through wastewater treatment plants, raising the question of their impact on human and environmental health. This has prompted the launch of several monitoring studies into the most commonly administered compounds in urban wastewater. The aim of this study was, therefore, to explore the cytotoxic potential of wastewaters samples collected from seven hospitals in Tunisia. The physicochemical analyses showed a large fluctuation of certain parameters in the collected samples, such as chemical oxygen demand (ranged from 860 to 1720 mg L-1), biochemical oxygen demand (ranged from 385 to 747 mg L-1), total organic carbon (ranged from 256 to 562 g L-1), total suspended solids (ranged from 905 to 1450 mg L-1), conductivity (ranged from 3.31 to 7.14 µsm/cm), and turbidity (ranged from 100 to 480 NTU). The analysis using inductively coupled plasma mass spectrometry (ICP-MS) also showed that hospital wastewater contains high concentrations of Hg (ranged from 0.0024 to 0.019 mg L-1). This could be explained by the variation of the activity and the services in certain hospitals compared to others. All hospital wastewater samples induced the proliferation of human breast cancer cell line MDA-231, even at low concentrations (20 µL/assay). Moreover, the maximum induction reached at the concentration of 60 µL/assay in wastewater samples from hospitals located in Monastir, Sidi Bouzid, Mahdia, and Sfax with percentages of induction up to 42.33, 14, 7.61, and 5.42%, respectively. These observations could be due to the presence of endocrine disrupting compounds (EDCs) in these wastewaters. Given this, our results evidenced the potential risk of these hospital effluents to environmental and public health.