Pharmaceuticals and personal care products in Tunisian hospital wastewater: occurrence and environmental risk.

Despite concerns about the potential risk associated with the environmental occurrence of pharmaceuticals and personal care products (PPCPs), few studies address the emissions of hospitals to aquatic compartments. We examined within a 3-month sampling period the occurrence and environmental risk of PPCPs in seven Tunisian hospital wastewaters. From personal care products, UV filters, main metabolites, and benzotriazoles were quantified, with benzophenone 3 (oxybenzone, BP3) and benzotriazole (BZT) the most frequently found (71%) at median concentrations in the range 2.43 ± 0.87 ngL-1-64.05 ± 6.82 ngL-1 for BP3 and 51.67 ± 1.67 ngL-1-254 ± 9.9 ngL-1 for BZT. High concentrations were also found for 4-hydroxybenzophenone (4HB) (221 ± 6.22 ngL-1), one of the main metabolites of BP3. The antibiotics ofloxacin and trimethoprim, the anti-inflammatory acetaminophen, the antiepileptic carbamazepine, and the stimulant caffeine were present in all the wastewaters. The highest median concentration corresponded to acetaminophen, with 1240 ± 94 mgL-1 in Tunis Hospital, followed by ofloxacin with 78850 ± 39 µgL-1 in Sousse Hospital. For ecotoxicity assessment, acute toxicity was observed for Daphnia magna and Vibrio fischeri. The toxicity data were used in a hazard quotient (HQ) approach to evaluate the risk posed by the target PPCPs to aquatic organisms. The calculated HQs revealed that marbofloxacin (234 for V. fischeri), enrofloxacin (121 for D. magna), and BZT (82.2 for D. magna and 83.7 for V. fischeri) posed the highest risk, concluding that potential risk exists toward aquatic microorganisms. This study constitutes the first monitoring of UV filters in Tunisian hospital effluents and provides occurrence and toxicity data of PPCPs for reference in further surveys in the country.

Single and combined effects of caffeine and salicylic acid on mussel Mytilus galloprovincialis: Changes at histomorphological, molecular and biochemical levels.

Caffeine (CAF) and salicylic acid (SA) are frequently detected in waterbody, though information on their biological impact is poor. This work assesses the effects of CAF (5 ng/L to 10 µg/L) and SA (0.05 µg/L to 100 µg/L) alone and combined as CAF+SA (5 ng/L+0.05 µg/L to 10 µg/L+100 µg/L) on mussel Mytilus galloprovincialis under 12-days exposure by histomorphology of digestive gland and oxidative stress defense at molecular and biochemical levels. Besides evaluating tissue accumulation, absence of histomorphological damage and haemocyte infiltration highlighted activation of defensive mechanisms. Up-regulation of Cu/Zn-sod, Mn-sod, cat and gst combined with increased catalase and glutathione S-transferase activity were found in CAF-exposed mussels, while SA reduced ROS production and mitochondrial activity. CAF+SA exposure induced differential responses, and the integrated biomarker response (IBR) revealed more pronounced effects of SA than CAF. These results enlarge knowledge on pharmaceuticals impact on non-target organisms, emphasizing the need for proper environmental risk assessment.

Identification of Novel Compounds Inhibiting the Kinase Activity of the CDK5/p25 Complex via Direct Binding to p25.

Tamoxifen, the gold standard drug for endocrine therapy for breast cancer, modulates the phosphorylation status of the TAU protein in Alzheimer's disease by inhibiting CDK5 kinase activity. Its binding to p25 prevents CDK5/p25 complexation and hence a decrease of CDK5 activity. In breast tumors, this complex is involved in the proliferation and survival of cancer cells, as well as in the disease's prognosis. Still, the molecular stability of the CDK5/p25 complex following tamoxifen exposure in this cancer type has not yet been clearly deciphered. Here, we report the functional characterization of CDK5 and its p25 regulatory subunit in the absence and presence of tamoxifen. In addition, two novel inhibitors of the kinase activity of the CDK5/p25 complex are identified, both of which would reduce the risk of recurrence of estrogen receptor-positive (ER+) breast cancers and prevent drawbacks induced by tamoxifen exposure. Accordingly, 6His-CDK5 and 6His-p25 have been expressed and purified. Fluorescence anisotropy measurements have been used to assess that the two proteins do form an active complex, and thermodynamic parameters of their interaction were measured. It was also confirmed that tamoxifen directly binds to p25 and inhibits CDK5 kinase activity. Similar observations were obtained using 4-hydroxytamoxifen, an active metabolized form of tamoxifen. Two novel compounds have been identified here that harbor a benzofuran moiety and were shown to target directly p25, and their bindings resulted in decreased CDK5 kinase activity. This encouraging alternative opens the way to the ensuing chemical optimization of this scaffold. It also promises a more specific therapeutic approach that may both tackle the pathological signaling in breast cancer and provide a potential new drug for Alzheimer's disease.

Occurrence and antibiotic resistance of Vibrio parahaemolyticus isolated from the Tunisian coastal seawater.

Vibrio parahaemolyticus is a gram-negative bacterium ubiquitous in seawater or estuarine water throughout the world. It is a major cause of seafood gastroenteritis complications. In this study, the presence of V. parahaemolyticus was investigated in 66 seawater samples collected during 2018 from 15 stations spread along the Tunisian coast using selective media including CHROMagar Vibrio media. The results show that only eight samples contained V. parahaemolyticus. However, while Vibrio alginolyticus was detected in all samples; both Vibrio cholerae and Vibrio vulnificus were not found. Nine of the presumed V. parahaemolyticus colonies were purified on tryptic soy agar from eight positive samples then identified by the API 20E biochemical test and confirmed by the presence of a specific target toxR gene. The detection of virulence genes, thermostable direct haemolysin (tdh) and thermostable-related haemolysin (trh), by the polymerase chain reaction (PCR) showed the presence of only two trh-positive isolates. The assessment of antibiotic susceptibility of the V. parahaemolyticus isolated revealed a complete resistance to colistin, amikacin, penicillin and cefotaxime and a total sensitivity to chloramphenicol, nitrofurantoin and sulfamethoxazole-trimethoprim with a multiple antibiotic resistance index (MAR) ranging from 0.4 to 0.5.

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.

Contaminants of emerging concern in marine areas: First evidence of UV filters and paraben preservatives in seawater and sediment on the eastern coast of Tunisia.

UV filters (UVFs) and paraben preservatives (PBs) are widely used components in many personal care products. However, there has been a rising concern for their endocrine-disrupting effects on wildlife once they reach aquatic ecosystems via recreative activities and wastewater treatment plants effluents. This study addresses UVFs and PBs occurrence in seawater and sediment impacted by tourism and sewage discharges along the coast of Mahdia, center East Tunisia. Samples of water and sediment were collected for 6 months from 3 coastal areas. Among the 14 investigated UVFs, 8 were detected in seawater and 4 were found in sediment. All PBs were present in seawater and only methylparaben (MePB) was detected in sediment. Benzophenone-3 (oxybenzone, BP3), benzocaine (EtPABA), and MePB were present in all water samples with concentrations in the ranges 16.4-66.9, 7.3-37.7, and 17.6-222 ng/L, respectively. However, the highest value, 1420 ng/L, corresponded to octinoxate (EHMC). In sediments, avobenzone (AVO), 4-methyl benzylidene camphor (4MBC), EHMC, 5-methyl-1-H-benzotriazole (MeBZT), and MePB were detected at concentrations within the range 1.1-17.6 ng/g dw, being MePB the most frequently detected (89%). MePB and MBZT presented the highest sediment-water partition coefficients and MePB also showed a positive correlation with total suspended solids' water content. Overall, pollutants concentrations remained rather constant along the sampling period, showing little seasonal variation. This study constitutes the first monitoring of UVFs and PBs on the Tunisian coastline and provides occurrence data for reference in further surveys in the country.

Time- and dose-dependent biological effects of a sub-chronic exposure to realistic doses of salicylic acid in the gills of mussel Mytilus galloprovincialis.

Among nonsteroidal anti-inflammatory drugs (NSAIDs) commonly found in seawater and wastewater, salicylic acid (SA) represents one of the most persistent and hazardous compounds for aquatic organisms. This study was therefore designed to elucidate the biological effects of SA in mussel Mytilus galloprovincialis. During a sub-chronic exposure (12 days), mussels were exposed to five realistic concentrations of SA (C1: 0.05 µg/L; C2: 0.5 µg/L; C3: 5 µg/L; C4: 50 µg/L; C5: 100 µg/L) and gills, selected as the target organ, were collected at different time points (T3: 3 days; T5: 5 days; T12: 12 days). Exposure to SA induced no histological alterations in mussel gills, despite a relevant hemocyte infiltration was observed throughout the exposure as a defensive response to SA. Temporal modulation of glutathione S-transferase (GST), catalase (CAT), and superoxide dismutase (SOD) activities suggested the occurrence of antioxidant and detoxifying responses against SA exposure, while lipid peroxidation (LPO), except for a partial increase at T3, was prevented. Inhibition of the cholinergic system was also reported by reduced acetylcholinesterase (AChE) activity, mainly at T12. Overall, findings from this study contribute to enlarge the current knowledge on the cytotoxicity of SA, on non-target aquatic organisms, and might for the enhancement of new ecopharmacovigilance programs and optimization of the efficacy of wastewater treatment plants for mitigation of pharmaceutical pollution in coastal areas.

Comparison of cellular mechanisms induced by pharmaceutical exposure to caffeine and its combination with salicylic acid in mussel Mytilus galloprovincialis.

Urban and hospital-sourced pharmaceuticals are continuously discharged into aquatic environments, threatening biota. To date, their impact as single compounds has been widely investigated, whereas few information exists on their effects as mixtures. We assessed the time-dependent biological impact induced by environmental concentrations of caffeine alone (CAF; 5 ng/L to 10 µg/L) and its combination with salicylic acid (CAF+SA; 5 ng/L+0.05 µg/L to 10 µg/L+100 µg/L) on gills of mussel Mytilus galloprovincialis during a 12-day exposure. Although no histological alteration was observed in mussel gills, haemocyte infiltration was noticed at T12 following CAF+SA exposure, as confirmed by flow cytometry with increased hyalinocytes. Both the treatments induced lipid peroxidation and cholinergic neurotoxicity, which the antioxidant system was unable to counteract. We have highlighted the biological risks posed by pharmaceuticals on biota under environmental scenarios, contributing to the enhancement of ecopharmacovigilance programmes and amelioration of the efficacy of wastewater treatment plants.

Histological endpoints and oxidative stress transcriptional responses in the Mediterranean mussel Mytilus galloprovincialis exposed to realistic doses of salicylic acid.

Despite the availability of analytic data, little is known about the toxicity of salicylic acid (SA) on aquatic non-target organisms. The present study aimed at evaluating the impact of SA through a short-term exposure of the Mediterranean mussel Mytilus galloprovincialis to five environmentally relevant concentrations of SA. A set of suitable biomarkers was applied at selected time-points on mussel digestive glands, including histological observations and expression of oxidative stress related genes. The obtained results showed a conspicuous hemocytic infiltration among mussel digestive tubules, as confirmed also by a flow cytometric approach that revealed an increase of halinocytes and granulocytes. Interestingly, a significant dose and time dependent decrease in the expression levels of oxidative stress related genes was found in mussels exposed to SA except for the glutathione S-transferase gene that was significantly up-regulated in a time-dependent manner confirming its important role against oxidant species and in the metabolism of pharmaceuticals.

Multidrug-resistant epi-endophytic bacterial community in Posidonia oceanica of Mahdia coast as biomonitoring factor for antibiotic contamination.

Faced with the significant disturbances, mainly of anthropogenic origin, which affect the Mediterranean coastal ecosystem, Posidonia oceanica (L.) Delile has often been used to assess the state of health of this environment. The present study aims to determine the multidrug resistance patterns among isolated and identified epi-endophytic bacterial strains in P. oceanica seagrass collected from Mahdia coastal seawater (Tunisia). To investigate the bacterial community structure and diversity from coastal seawater samples from Mahdia, total DNA extraction and 16S rRNA gene amplification were performed and analyzed by denaturing gradient gel electrophoresis (DGGE). The DGGE profiles showed that some bands were specific to a given site, while other bands were found to be common to more than one sample. In the other hand, bacterial strains were isolated from 1 mL of leaves and epiphytes suspension of P. oceanica seagrass in marine agar. Forty-three isolates were obtained, seven of them were selected and identified on the basis of 16S rRNA gene sequence analysis. These isolates belonged to the genus Bacillus, exhibiting 98-100% of identity with known sequences. Susceptibility patterns of these strains were studied toward commonly used antibiotics in Tunisia. All identified isolates were resistant to Aztreonam (72.1%), Ceftazidime (60.5%), Amoxicillin (56%) and Rifampicin (51.2%). S5-L13 strain had presented the highest multidrug resistance with a MAR index of 0.67.

Chemical Characterization of Different Products from the Tunisian Opuntia ficus-indica (L.) Mill.

Various dried (by-)products from the Tunisian O. ficus-indica were elucidated for their proximate composition, fatty acid (FA) composition, inorganic elements, sugars, and polyphenols. Nopal and prickly pear peel and seeds were abundant in fiber (respectively, 28.39, 12.54, and 16.28%). Seeds had also high protein (17.34%) and may be source of an edible oil, due to lipids (9.65%) poor in saturated FAs (14.12%) and rich in linoleic acid (61.11%). Nopal and peel showed the highest levels of Mg (493.57 and 345.19 mg/100 g), K (6949.57 and 1820.83 mg/100 g), Mn (59.73 and 46.86 mg/Kg) and Fe (23.15 and 15.23 mg/Kg), while the fruit pulp predominantly constituted of sugars, glucose and arabinose being predominant (42.57 and 13.56 g/100 g). Total polyphenols widely varied among the Opuntia products (108.36-4785.36 mg GAE/100 g), being mainly represented by hydroxycinnamic and hydroxybenzoic acids, and flavonoids as well. In particular, peel may be revalorized for these valuable bioactives, including 4-hydroxybenzoic acid (484.95 mg/100 g), cinnamic acid (318.95 mg/100 g), rutin (818.94 mg/100 g), quercetin (605.28 mg/100 g), and several isorhamnetin and kaempferol glycosides. Overall, the Tunisian prickly pear cactus could encourage a sustainable production, an effective waste management, and may provide several benefits for human health, in accordance with the model of the Mediterranean diet.

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.

Occurrence of Textile Dyes and Metals in Tunisian Textile Dyeing Effluent: Effects on Oxidative Stress Status and Histological Changes in Balb/c Mice.

Although it is known that textile wastewater contains highly toxic contaminants whose effects in humans represent public health problems in several countries, studies involving mammal species are scarce. This study was aimed to evaluate the toxicity profile of 90-days oral administration of textile dyeing effluent (TDE) on oxidative stress status and histological changes of male mice. The TDE was collected from the textile plant of Monastir, Tunisia and evaluated for the metals, aromatic amines, and textile dyes using analytical approaches. Metal analysis by ICP-MS showed that the tested TDE exhibited very high levels of Cr, As, and Sr, which exceeded the wastewater emission limits prescribed by WHO and Tunisian authority. The screening of TDE through UPLC-MS/MS confirmed the presence of two textile dyes: a triphenylmethane dye (Crystal violet) and a disperse azo dye (Disperse yellow 3). Exposure to TDE significantly altered the malondialdehyde (MDA), Conjugated dienes (CDs), Sulfhydryl proteins (SHP) and catalase levels in the hepatic and renal tissues. Furthermore, histopathology observation showed that hepatocellular and renal lesions were induced by TDE exposure. The present study concluded that TDE may involve induction of oxidative stress which ensues in pathological lesions in several vital organs suggesting its high toxicity. Metals and textile dyes may be associated with the observed toxicological effects of the TDE. These pollutants, which may have seeped into surrounding rivers in Monastir city, can cause severe health malaise in wildlife and humans.

Factors controlling phytoplankton dynamics in an arid reservoir in Tunisia (case of Sidi Saad dam).

Temporal variation of the phytoplankton community and environmental factors were investigated over 8 months: from January 2018 to November 2018 in the Sidi Saad reservoir (central Tunisia): May, June (spring season), July, August (summer season) September, October and November (autumn season), and January (winter season). The relationships between phytoplankton and environmental factors were explored using Canonical Correspondence Analysis (CCA). General linear models (GLMs) were used to predict the phytoplankton abundance. Analysis of variance (ANOVA) was used to test the hypothesis that the abundance of each group of taxa differed between sampling months. The results of chemical analysis of the reservoir showed that the environment was enriched in nitrates, nitrites, ammonium, and orthophosphate, especially in May. Carlson's trophic index using average chemical variables showed that Sidi Saad has a mesotrophic statute. There is temporal phytoplankton succession in the Sidi Saad reservoir. Cyanophyceae dominated over the year except in winter and late autumn (November and January). Chlorophyceae was the dominant group in winter month. The CCA results showed that six environmental factors, orthophosphates (PO43-), total phosphates (TP), Secchi disc depth (SD), total nitrogen (TN), temperature (T), and nitrite (NO2-) had significant influences on the changes in phytoplankton. GLM showed that PO43-, TP, TN, SD, and T were the significant predictors of phytoplankton abundance. Phytoplankton composition is largely dominated by the species Microcystis aeruginosa which formed a bloom with excessive abundance (up to 89.76 billion cell l-1 in spring). We recommend banning the fishing and their consumption during the period of Microcystis bloom and installing a system of biomonitoring of the levels of toxins in the water.

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.

Virulence of clinically relevant multidrug resistant Corynebacterium striatum strains and their ability to adhere to human epithelial cells and inert surfaces.

Corynebacterium striatum is a nosocomial pathogen which is increasingly associated with serious infections in both immunocompetent and immunocompromised patients. However, little is known about virulence factors and mechanisms that may enhance the establishment and long-term survival of Corynebacterium striatum. in the hospital environment. In this study, we investigated the ability of 22 multidrug-resistant C. striatum clinical isolates to adhere to human epithelial cells and to produce biofilm on polystyrene plates, glass and various tracheostomy tubes. We also tested the virulence of these strains on the nematode Caenorhabditis elegans. They showed good adhesion to epithelial human cells after 180 min of infection. The 22 C. striatum were able to produce biofilms on positively and negatively charged abiotic surfaces at 37 °C. They were also able to infect and to kill Caenorhabditis elegans after 5 days of infection. The virulence condition was associated with the presence of SpaDEF operon encoding pili in all strains. This study provides new insights on virulence mechanisms that may contribute to the persistence of C. striatum in the hospital environment, increasing the probability of causing nosocomial infections.

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.

Organic pollutants in marine samples from Tunisian coast: Occurrence and associated human health risks.

140 contaminants belonging to various classes (organochlorine and organophosphorus pesticides, pyrethroid insecticides, carbamates, fungicides, acaricides, herbicides, synergists, insect growth regulators, polychlorobiphenyls, polycyclic aromatic hydrocarbons) were simultaneously analysed by GC-MS/MS in marine sediments, aquatic plant leaves and fish tissues samples. A total of 260 samples from five stations along the coast of Tunisia were evaluated. The results highlight that only 28 residues (12 polychlorobiphenyls, 8 organochlorine pesticides, 7 polycyclic aromatic hydrocarbons and triphenyl phosphate) were detected at levels higher than relative LOQ values. The amounts in sediment samples were compared with Sediment Quality Guidelines (SQGs) showing that the values are acceptable and no toxic effect is expected on aquatic organisms. A little variation of contaminant residues in sediment samples among coastal stations was recorded. Namely, with respect to almost all polychlorobiphenyls and organochlorine pesticides, higher values were recorder in summer. With respect to almost all polycyclic aromatic hydrocarbons, higher values were recorder in autumn. Aquatic plant leaves showed a residue accumulation higher than that of other compartments of marine system. The data about fish samples (Sparus aurata and Sarpa salpa, the two most frequently caught fish species at five sites on the central coast of Tunisia) do not pose direct hazard to human health because values were lower than protection limits.

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.

Anti-oxidant, antibacterial, anti-biofilm, and anti-quorum sensing activities of four essential oils against multidrug-resistant bacterial clinical isolates.

PURPOSE OF THE STUDY: Outbreaks of multidrug-resistant bacteria are increasingly reported at the clinical setting. The antimicrobial, anti-biofilm, anti-quorum sensing, and anti-oxidant activities of four essential oils extracted from Cinnamomum verum, Origanum majorana, Thymus vulgaris, and Eugenia caryophyllata against Gram-positive and Gram-negative multidrug-resistant bacteria were evaluated in vitro. MATERIALS AND METHODS: This study was conducted on 105 multidrug resistant clinical strains. Inhibition diameter zone, minimum inhibitory concentration, and minimum bactericide concentration of the oils were determined using agar disc diffusion method and microdilution. The ability of the 4 essential oils to inhibit the production of bacterial biofilms was tested on polystyrene plates, as well as their inhibitory effect on the production of violacein by Chromobacterium violaceum CV026. The anti-oxidant activity was evaluated by the 2,2-diphenyl-1-picrylhydrazyl scavenging method. RESULTS: Essential oils of Cinnamomum verum, Thymus vulgaris and Eugenia caryophyllata showed an important antibacterial activity. The inhibition diameter zone was higher than 20 mm for 90.24 %, 85.71 % and 60.95 % of strains respectively. These essential oils have a remarkable anti-biofilm and anti-quorum sensing activities against almost all the species studied. Clove extract revealed the highest anti-oxidant activity (Pourcentage of inhibtion of DPPH = 90.3 %). CONCLUSION: These results supported the use of the 4 essential oils as alternative or complementary agents to treat infections caused by multidrug-resistant bacteria, and to prevent biofilm formation and quorum sensing signaling. They might be used as a safe anti-oxidants instead of harmful artificial ones.