Lipopeptide culture filtrates from Bacillus spp. provide effective protection to wheat against the foliar pathogen Zymoseptoria tritici.

AIMS: Biocontrol products based on microorganisms and natural substances are promising alternatives to chemical pesticides that could contribute to develop a more sustainable agriculture. Here, we investigated the potential of cell-free culture filtrates (CFCFs) from two strains of the Bacillus subtilis group to inhibit Zymoseptoria tritici, a major fungal pathogen of wheat. METHODS AND RESULTS: Foliar application of CFCFs from Bacillus velezensis GA1 and Bacillus sp. III1 on wheat seedlings in a greenhouse strongly reduced Z. tritici disease severity (>90%). In vitro bioassays showed that CFCFs completely inhibited the spore germination and fungal growth (100%). In planta cytological investigations revealed a significant impact of the treatments on both spore germination (∼40% inhibition) and fungal growth of Z. tritici (>80% inhibition). High Performance Liquid Chromatography (HPLC) analysis showed that the Bacillus strains displayed different lipopeptide profiles. The CFCF obtained from Bacillus GA1 contained 90 mg l-1 of iturin A + surfactins + fengycins and the CFCF obtained from Bacillus sp. III1 contained 25 mg l-1 of mojavensin A (iturin family) + surfactins + fengycins. CONCLUSIONS: Strains of the B. subtilis group producing different iturins could provide several CFCF-based solutions for the biocontrol of Z. tritici.

Potential benefits of polysaccharides derived from marine alga Ulva lactuca against hepatotoxicity and nephrotoxicity induced by thiacloprid, an insecticide pollutant.

The present study aimed to evaluate the potential protective and antioxidant effect of polysaccharides (PS) extracted from Ulva lactuca against thiacloprid (THC) induced nephrotoxicity and hepatotoxicity. The antioxidant capacity of PS was tested in vitro using ABTS radical scavenging activity and plasmid DNA cleavage assays andin vivo on adult male rats treated for 30 days. Animals were allocated into four groups: control; THC (22.5 mg/kg); THC (22.5 mg/kg) + PS1 (100 mg/kg diet); and THC (22.5 mg/kg) + PS2 (200 mg/kg diet). The structural features of PS were determined by Fourier transformed infrared (FT-IR), UV absorption peak detection, high performance liquid chromatography (HPLC) and gel permeation chromatography, and also functional properties were investigated. Overall, results indicated that THC increased significantly malondialdehyde, advanced oxidation protein products, glutathione levels, which is correlated with severe histological and plasmatic biochemical injuries in both liver and kidney tissues. However, cotreatment PS induced a significant protective and healing affects against the nephrotoxicity and hepatotoxcity induced by THC.

Chemical characterization and biological activities of Simmondsia chinensis (Link) C. K. Schneid seeds oil.

The chemical composition, main physicochemical properties, and biological activities of Simmondsia chinensis (S. chinensis) seeds oil were studied. The results revealed that the physiochemical characteristics of S. chinensis seeds oil were as follows: acid values 1.15 mg KOH/g, peroxide values 8.00 meq O2 Kg-1, iodine values 80.00 g/100 g of oil and saponification values 92.00 mg KOH/g, phenolic content 50.91 mg gallic acid equivalents/g extract. Gas chromatography analysis indicated that eicosenoic (55.50 %), erucic (20.43 %) and oleic (19.01 %) acids were the most abundant, saturated and unsaturated, fatty acids in the oil. Moreover, the evaluation of their antioxidant (DPPH, TAC), antibacterial, antidiabetic and acetylcholinesterase evinced interesting results. Seeds of S. chinensis constitute a substitute source for stable vegetable oil and protein with regard to nutritional and industrial applications.

Date Palm Trees Root-Derived Endophytes as Fungal Cell Factories for Diverse Bioactive Metabolites.

Endophytic fungi of healthy and brittle leaf diseased (BLD) date palm trees (Phoenix dactylifera L.) represent a promising source of bioactive compounds with biomedical, industrial, and pharmaceutical applications. The fungal endophytes Penicillium citrinum isolate TDPEF34, and Geotrichum candidum isolate TDPEF20 from healthy and BLD date palm trees, respectively, proved very effective in confrontation assays against three pathogenic bacteria, including two Gram-positive bacteria Bacillus thuringiensis (Bt) and Enterococcus faecalis (Ef), and one Gram-negative bacterium Salmonella enterica (St). They also inhibited the growth of three fungi Trichoderma sp. (Ti), Fusarium sporotrichioides (Fs), Trichoderma sp. (Ts). Additionally, their volatile organic compounds (VOCs) were shown to be in part responsible for the inhibition of Ti and Ts and could account for the full inhibition of Fs. Therefore, we have explored their potential as fungal cell factories for bioactive metabolites production. Four extracts of each endophyte were prepared using different solvent polarities, ethanol (EtOH), ethyl acetate (EtOAc), hexane (Hex), and methanol (MetOH). Both endophyte species showed varying degrees of inhibition of the bacterial and fungal pathogens according to the solvent used. These results suggest a good relationship between fungal bioactivities and their produced secondary metabolites. Targeting the discovery of potential anti-diabetic, anti-hemolysis, anti-inflammatory, anti-obesity, and cytotoxic activities, endophytic extracts showed promising results. The EtOAc extract of G. candidum displayed IC50 value comparable to the positive control diclofenac sodium in the anti-inflammatory assays. Antioxidant activity was evaluated using α,α-diphenyl-ß-picrylhydrazyl (DPPH), ß-carotene bleaching, reducing power (RP), and 2,2-azino-bis(3-ethylbenzothiazoline-6-sulphonique) (ABTS) radical scavenging assays. The findings revealed strong anti-oxidant power with an IC50 of 177.55 µg/mL for G. candidum EtOAc extract using DPPH assay, probably due to high polyphenol and flavonoid content in both fungal extracts. Finally, LC-HRMS (Liquid Chromatography­High Resolution Mass Spectrometry) and GC-MS (Gas Chromatography­Mass Spectrometry) analysis of G. candidum and P. citrinum extracts revealed an impressive arsenal of compounds with previously reported biological activities, partly explaining the obtained results. Finally, LC-HRMS analysis indicated the presence of new fungal metabolites that have never been reported, which represent good candidates to follow for the discovery of new bioactive molecules.

Anti-microbial screening and cytotoxic activity of aerial part of Thymelaea hirsuta L. essential oil growing in south-west Tunisia.

This study aimed to investigate the antimicrobial and cytotoxic activities of essential oil isolated by the hydro-distillation of aerial parts of Thymelaea hirsuta. The antimicrobial activity of the oil was evaluated against eight bacterial and three fungal pathogenic strains. The results revealed that the essential oil exhibited a moderate-to-potent anti-microbial activity against all the microorganisms tested. Gram-positive bacteria were noted to be more sensitive to the oil than gram-negative bacteria and yeasts. In vitro cytotoxicity evaluation against HeLa cell lines showed that the essential oil exhibited moderate cytotoxicity on human tumor cells, with a high IC50 value of 175µg/mL. To the author's knowledge, this is the first study reporting on the antimicrobial and cytotoxic activities of Thymelaea hirsuta essential oil. Overall, the results indicate that the T. hirsuta essential oil has a number of attractive properties that might open new promising opportunities for the control or prevention of a wide range of microbial infections and cancers and can facilitate the use of essential oils as natural preservatives against spoilage microorganisms in food systems.

Chemical composition and in vitro evaluation of antioxidant, antimicrobial, cytotoxicity and anti-acetylcholinesterase properties of Tunisian Origanum majorana L. essential oil.

This study investigated the chemical composition and evaluated the antioxidant, antimicrobial, cytotoxic and anti-acetylcholinesterase properties of Tunisian Origanum majorana essential oil. The findings showed that the oil exhibited high activity, particularly in terms of reducing power and ß-Carotene bleaching, inducing higher IC50 values than BHT. The oil showed an important antimicrobial activity against 25 bacterial and fungal strains. In fact, the IZ, MIC and MBC values recorded for the bacterial strains were in the range of 8 ± 0-18.33 ± 0.57 mm, 0.097-3.125 and 0.39-6.25 mg/mL, respectively. The IZ, MIC and MFC values of the fungal strains varied between 11±0-28 ± 0 mm, 0.058-0.468 mg/mL and 0.234-1.875 mg/mL, respectively. A low cytotoxic effect was observed against cancer (Hep-2 and HT29) and continuous cell lineage (Vero), with CC50 values ranging from 13.73 to 85.63 mg/mL. The oil was also evaluated for anti-acetylcholinesterase effects, which showed that it exhibited significant activity with IC50 values reaching 150.33 ± 2.02 µg/mL.

Essential oil of Zygophyllum album inhibits key-digestive enzymes related to diabetes and hypertension and attenuates symptoms of diarrhea in alloxan-induced diabetic rats.

CONTEXT: Zygophyllum album L. (Zygophyllaceae), commonly known as Bougriba, is widely used to treat diabetes, digestive tract spasm, and hypertension in folk medicine, in Tunisia. OBJECTIVE: This study investigates the antidiabetic, antidiarrheal, and antihypertensive activities of the leaves of the essential oil from Zygophyllum album (OZA) in alloxan-induced diabetic rats. MATERIALS AND METHODS: The oil was obtained by hydrodistillation and analyzed by GC-MS. Males rats were divided into four groups: control, diabetic-untreated group, diabetic-treated group with acarbose (10 mg/kg), and diabetic-treated rats with OZA (200 mg/kg) for 30 d. RESULTS: At the end of the experimental period, the OZA significantly decreased the activity of α-amylase in pancreas and serum of the diabetic rats by 43% and 38%, respectively, which led to reduce the serum glucose level by 60% and lower of glycated hemoglobin (HbA1c) rate by 17% as compared with untreated diabetic animals. Moreover, the OZA treatment attenuated symptoms of diarrhea, improved lipid disorders, and hypertension through inhibiting the pancreatic lipase and angiotensin-converting enzyme (ACE) activities by 47% and 25%, respectively, in serum of diabetic rats. CONCLUSION: OZA showed a good effect in the management of diabetes mellitus and exerted preventive action from related hypertension.

Preventive effects of oleuropein against cardiac remodeling after myocardial infarction in Wistar rat through inhibiting angiotensin-converting enzyme activity.

OBJECTIVE: Myocardial infarction remains the major cause of global death due to cardiovascular diseases. This study aimed to assess the protective role of oleuropein in attenuating the cardiac remodeling in isoproterenol-induced myocardial infarction in rats. METHODS AND RESULTS: Male Wistar rats were randomly divided into four groups, control, isoproterenol (Isop) and pretreated animals with oleuropein at two different doses (20 and 40 mg/kg) orally for 7 days and intoxicated with isoproterenol (Isop+Oleu20) and (Isop+Oleu40) groups. The subcutaneous injection of isoproterenol (100 mg/kg body weight) to untreated rats for two consecutive days showed significant increases in ST-segment elevation, heart weight index and alteration in the ECG pattern and hemodynamic function. Else, serum levels of cardiac troponin-T, creatine kinase isoenzyme (CK-MB), lactate dehydrogenase (LDH) and alanine aminotransferase (ALT) underwent a notable rise in serum of Isop group by (345, 82, 73 and 106%, respectively) as compared to normal rats. Isoproterenol-induced myocardial injury was evidenced by alteration in serum lipids profile and increased activities of pancreatic lipase by 94% and angiotensin-converting enzyme (ACE) by 78% which reflects the occurrence of cardiac remodeling process. The histopathological findings of the infarcted group showed myocardium necrosis and cells inflammatory infiltration. However, the treatment with oleuropein gave a good protection of the myocardium by decreasing cardiac injury markers specially troponin-T, restoring hemodynamic parameters and attenuating cardiac remodeling process through inhibition of ACE activity. CONCLUSION: Oleuropein offers high preventive effects from cardiac remodeling process in rats with acute myocardial infarction.

Anti-obesity and cardioprotective effects of cinnamic acid in high fat diet- induced obese rats.

Obesity is a chronic metabolic disorder that is associated with numerous diseases including hyperlipidemia, diabetes mellitus, hypertension, atherosclerosis, cardiovascular disease, and cancer. Cinnamic acid is a phytochemical compound having many biological effects and could be considered for the management of obesity. This study is aimed to assess the possible anti-obesity and cardioprotective properties of cinnamic acid (CA) in high fat diet-fed rats (HFD). Male Wistar rats were divided into 4 groups. They received normal diet, HFD diet, HFD supplemented with fluvastatin (2 mg/kg/day) or cinnamic acid (30 mg/kg/day) for 7 weeks. The results showed an increase in body weight of HFD rats by ~27 % as compared to control group. Moreover, serum lipase activity underwent a significant rise by 103 % which led to an increase in the levels of total cholesterol (T-Ch), triglycerides (TG), LDL-cholesterol in serum of untreated HFD-fed rats. Furthermore, the concentration of leptin and angiotensin-converting enzyme (ACE) activity exhibited remarkable increases in serum of HFD-fed rats as compared to controls. Whereas, the administration of CA to HFD-fed rats improved the body weight gain and serum lipid profile and reverted back near to normal the activities of lipase and ACE. In addition, the echocardiography evidenced that CA is able to protect the aorta and aortic arch and avoided vasoconstriction by increasing their diameters and improved liver steatosis and kidney indices of toxicity. Overall, these results suggest that cinnamic acid exerts anti-obesity and antihypertensive effects through inhibition of lipid digestive enzymes and ACE.

Isolation of a novel amylase and lipase-producing Pseudomonas luteola strain: study of amylase production conditions.

An amylase and lipase producing bacterium (strain C2) was enriched and isolated from soil regularly contaminated with olive washing wastewater in Sfax, Tunisia. Cell was aerobic, mesophilic, Gram-negative, motile, non-sporulating bacterium, capable of growing optimally at pH 7 and 30°C and tolerated maximally 10% (W/V) NaCl. The predominant fatty acids were found to be C(18:1)ω7c (32.8%), C(16:1)ω7c (27.3%) and C16:0 (23.1%). Phylogenetic analysis of the 16S rRNA gene revealed that this strain belonging to the genus Pseudomonas. Strain C2 was found to be closely related to Pseudomonas luteola with more than 99% of similarity. Amylase optimization extraction was carried out using Box Behnken Design (BBD). Its maximal activity was found when the pH and temperature ranged from 5.5 to 6.5 and from 33 to 37°C, respectively. Under these conditions, amylase activity was found to be about 9.48 U/ml.

Biological properties of carotenoids extracted from Halobacterium halobium isolated from a Tunisian solar saltern.

BACKGROUND: Bioactive molecules have received increasing attention due to their nutraceutical attributes and anticancer, antioxidant, antiproliferative and apoptosis-inducing properties. This study aimed to investigate the biological properties of carotenoids extracted from Archaea. METHODS: Halophilic Archaea strains were isolated from the brine of a local crystallizer pond (TS7) of a solar saltern at Sfax, Tunisia. The most carotenoid-producing strain (M8) was investigated on heptoma cell line (HepG2), and its viability was assessed by the MTT-test. The cells were incubated with different sub-lethal extract rates, with carotenoid concentrations ranging from 0.2 to 1.5 µM. Antioxidant activity was evaluated through exposing the cells to sub-lethal extract concentrations for 24 hours and then to oxidative stress induced by 60 µM arachidonic acid and 50 µM H2O2. RESULTS: Compared to non-treated cells, bacterial carotenoid extracts inhibited HepG2 cell viability (50%). A time and dose effect was observed, with cell viability undergoing a significant (P < 0.05) decrease with extract concentration. After exposure to oxidative stress, control cells underwent a significant (P < 0.05) decrease in viability as compared to the non-treated cells. CONCLUSIONS: The bacterial extracts under investigation were noted to exhibit the strongest free radical scavenging activity with high carotenoid concentrations. The carotenoid extract also showed significant antiproliferative activity against HepG2 human cancer cell lines.

Aspergillus niger P6 and Rhodotorula mucilaginosa CH4 used for olive mill wastewater (OMW) biological treatment in single pure and successive cultures.

The aim of this study was to investigate the Rhodotorula mucilaginosa CH4 and Aspergillus niger P6 abilities to purify olive mill wastewater (OMW) in single pure and mixed cultures during the treatment. Both fungi were molecularly identified. OMW was used at five dilutions from 5% to 30% with chemical oxygen demand (COD) ranging from 11,600 to 24,600 mg L(-1). Firstly, each fungus was used separately, then they were successively used to treat the OMW. In single pure culture, A. niger showed a better efficiency in OMW purification than R. mucilaginosa. Furthermore, when successively used, the two studied strains exhibited improvements in the decrease of COD, polyphenolic compounds concentration and effluent colour. COD removals were 95.68-56.71% by R. mucilaginosa and 98.02-69.51% by A. niger for OMW dilutions varying from 5% to 30%. Both strains showed an important polyphenolic compounds removal of 83-45% by R. mucilaginosa and 94-58% by A. niger, in accordance with the OMW COD initially used. The COD and phenolic compound removals fitted simple equation models, with high regression coefficients. The strains' growth kinetics decreased according to the OMW concentration, but, when successively used, fungal growth was improved, allowing efficient effluent treatment.

Influence of Zeolite on fatty acid composition and egg quality in Tunisian Laying Hens.

BACKGROUND: The health benefits of omega-3 and omega-6 polyunsaturated fatty acids (PUFA) are generally recognized. Unfortunately, in most Mediterranean countries, the recommended daily intake of these compounds is rarely met. Therefore, enrichment of commonly occurring foods can boost intake of these fatty acids. In this regard, eggs are an interesting target, as they form an integral part of the diet. RESULT: Zeolite (Clinoptilolites) was added to Laying Hens feed at concentrations 1% or 2% and was evaluated for its effects on performance of the production and on egg quality. The Laying Hens were given access to 110 g of feed mixtures daily that was either a basal diet or a 'zeolite diet' (the basal diet supplemented with clinoptilolite at a level of 1% or 2%). It was found that zeolite treatment had a positive and significatif (p<0.05) effect on some parameters that were measured like egg height and eggshell strength. While dietary zeolite supplementation tended to/or has no significant effects on total egg, eggshell, yolk and albumen weights. It was found also that zeolite mainly increases level of polyunsaturated fatty acids in egg. CONCLUSION: This study showed the significance of using zeolite, as a feed additive for Laying Hens, as part of a comprehensive program to control egg quality and to increase level of polyunsaturated fatty acids on egg.

Essential oil of the leaves of Ricinus communis L.: in vitro cytotoxicity and antimicrobial properties.

BACKGROUND: The aim of the present study was to appraise the antimicrobial activity of Ricinus communis L. essential oil against different pathogenic microorganisms and the cytotoxic activity against HeLa cell lines. METHODS: The agar disk diffusion method was used to study the antibacterial activity of Ricinus communis L. essential oil against 12 bacterial and 4 fungi strains. The disc diameters of zone of inhibition (DD), the minimum inhibitory concentrations (MIC) and the concentration inhibiting 50% (IC50) were investigated to characterize the antimicrobial activities of this essential oil. The in vitro cytotoxicity of Ricinus communis L. essential oil was examined using a modified MTT assay; the viability and the IC50 were used to evaluate this test. RESULTS: The essential oil from the leaves of Ricinus communis L. was analyzed by GC-MS and bioassays were carried out. Five constituents of the oil were identified by GC-MS. The antimicrobial activity of the oil was investigated in order to evaluate its efficacy against twelve bacteria and four fungi species, using disc diffusion and minimum inhibitory concentration methods. The essential oil showed strong antimicrobial activity against all microorganisms tested with higher sensitivity for Bacillus subtilis, Staphylococcus aureus and Enterobacter cloacae. The cytotoxic and apoptotic effects of the essential oil on HeLa cell lines were examined by MTT assay. The cytotoxicity of the oil was quite strong with IC50 values less than 2.63 mg/ml for both cell lines. CONCLUSION: The present study showed the potential antimicrobial and anticarcinogenic properties of the essential oil of Ricinus communis L., indicating the possibilities of its potential use in the formula of natural remedies for the topical treatment of infections.

Yeast performance in wastewater treatment: case study of Rhodotorula mucilaginosa.

The ability of Rhodotorula mucilaginosa to degrade some phenolic compounds and to grow on olive mill wastewater (OMW) is investigated. R. mucilaginosa isolated from OMW was molecularly identified using 18S RNA sequencing. The biodegradation of six phenolic compounds was studied at an initial concentration of 1 g l(-1). The isolated yeast exhibited a complete degradation of protocatechuic, vanillic and p-coumaric acids and tyrosol. In addition, it reduced 56% and 44% of gallic acid and catechol, respectively. Protocatechuic acid, vanillic acid and p-coumaric acid kinetic degradation showed a simple order equation and the growth rate varied from 0.05 h(-1) to 0.08 h(-1), while tyrosol and catechol degradation fitted a second-degree equation. With OMW as culture medium, R. mucilaginosa was able to reduce 38.38%, 47.69% and 56.91% of chemical oxygen demand (COD) and 5.84%, 27.89% and 34.81% of phenols, respectively, at initial COD concentrations 26,700, 14,400 and 6500 mg l(-1). The use of such red pigmented yeast would present a double interest: first it would purify OMW and, second, an antioxidant would be produced at the same time, having antioxidant properties.

The in-vitro evaluation of antibacterial, antifungal and cytotoxic properties of Marrubium vulgare L. essential oil grown in Tunisia.

BACKGROUND: In order to validate its antiseptic and anticancer properties with respect to traditional uses, we have screened for the first time the antimicrobial activity of aerial parts of M. vulgare L. essential oil against different pathogenic microorganisms and the cytotoxic activity against HeLa cell lines. METHODS: The agar disk diffusion method was used to study the antibacterial activity of M. vulgare essential oil against 12 bacterial and 4 fungi strains. The disc diameters of zone of inhibition (DD), the minimum inhibitory concentrations (MIC) and the concentration inhibiting 50% (IC50) were investigated to characterize the antimicrobial activities of this essential oil. The in vitro cytotoxicity of M. vulgare essential oil was examined using a modified MTT assay; the viability and the IC50 were used to evaluate this test. RESULTS: The antimicrobial activity of the essential oil was investigated in order to evaluate its efficacy against the different tested microorganisms. The present results results showed a significant activity against microorganisms especially Gram (+) bacteria with inhibition zones and minimal inhibitory concentration values in the range of 6.6-25.2 mm and 1120-2600 µg/ml, respectively, whereas Gram (-) bacteria exhibited a higher resistance. As far as the antifungal activity, among four strains tested, Botrytis cinerea exhibited the strongest activity with inhibition zones of 12.6 mm. However, Fusarium solani, Penicillium digitatum and Aspergillus niger were less sensitive to M. vulgare essential oil. About the citotoxicity assay, this finding indicate the capability of this essential oil to inhibited the proliferation of HeLa cell lines under some conditions with IC50 value of 0.258 µg/ml. CONCLUSION: This investigation showed that the M. vulgare essential oil has a potent antimicrobial activity against some Gram (+) pathogenic bacteria and Botrytis cinerea fungi. The present studies confirm the use of this essential oil as anticancer agent. Further research is required to evaluate the practical values of therapeutic applications.

Molecular community analysis of magnesium-rich bittern brine recovered from a Tunisian solar saltern.

The microbial community of a magnesium-rich bittern brine saturated with NaCl (380-400 g/L) from a Tunisian solar saltern was investigated using a molecular approach based on 16S rRNA gene analysis and viability tests. The results revealed the existence of microbial flora. Viability test assessment showed that 46.4% of this flora was viable but not detectable by culturability tests. 16S rRNA genes from 49 bacterial clones and 38 archaeal clones were sequenced and phylogenetically analyzed. Eleven operational taxonomic units (OTUs) determined by the DOTUR program with 97% sequence similarity were generated for Bacteria. These OTUs were affiliated with Bacteroidetes and Gammaproteobacteria. The archaeal community composition exhibited more diversity with 38 clones, resulting in 13 OTUs affiliated with the Euryarchaeota phylum. Diversity measurement showed a more diverse archaeal than bacterial community at the saturated pond.

Microbial community of salt crystals processed from Mediterranean seawater based on 16S rRNA analysis.

Phylogenetic analysis of 16S rRNA was used to investigate for the first time the structure of the microbial community that inhabits salt crystals retrieved from the bottom of a solar saltern, located in the coastal area of the Mediterranean Sea (Sfax, Tunisia). This community lives in an extremely salty environment of 250-310 g/L total dissolved salt. A total of 78 bacterial 16S rRNA clone sequences making up to 21 operational taxonomic units (OTUs), determined by the DOTUR program to 97% sequence similarity, was analyzed. These OTUs were affiliated to Bacteroidetes (71.4% of OTUs), and gamma-Proteobacteria and alpha-Proteobacteria (equally represented by 14.2% of the OTUs observed). The archaeal community composition appeared more diverse with 68 clones, resulting in 44 OTUs, all affiliated with the Euryarchaeota phylum. Of the bacterial and archaeal clones showing <97% 16S rRNA sequence identity with sequences in public databases, 47.6% and 84.1% respectively were novel clones. Both rarefaction curves and diversity measurements (Simpson, Shannon-Weaver, Chao) showed a more diverse archaeal than bacterial community at the Tunisian solar saltern pond. The analysis of an increasing clone's number may reveal additional local diversity.

Isolation and characterization of moderately halophilic bacteria from Tunisian solar saltern.

Bacterial screenings from solar saltern in Sfax (Tunisia) lead to the isolation of 40 moderately halophilic bacteria which were able to grow optimally in media with 5-15% of salt. These isolates were phylogenetically characterized using 16S rRNA gene sequencing. Two groups were identified including 36 strains of Gamma-Proteobacteria (90%) and 4 strains of Firmicutes (10%). The Gamma-Proteobacteria group consisted of several subgroups of the Halomonadaceae (52.5%), the Vibrionaceae (15%), the Alteromonadaceae (10%), the Idiomarinaceae (7.5%), and the Alcanivoracaceae (5%). Moreover, three novel species: 183ZD08, 191ZA02, and 191ZA09 were found, show <97% sequence similarity of the 16S rRNA sequences while compared to previously published cultivated species. Most of these strains (70%) were able to produce hydrolases: amylases, proteases, phosphatases, and DNAases. Over the isolates, 60% produced phosphatases, 15.0% proteases, 12.5% amylases and DNAases equally. This study showed that the solar saltern of Sfax is an optimal environment for halophilic bacterial growth, where diverse viable bacterial communities are available and may have many industrial applications.

Olive mill wastewater disposal in evaporation ponds in Sfax (Tunisia): moisture content effect on microbiological and physical chemical parameters.

The study of the isotherms desorption of olive mill wastewater (OMW) was investigated to describe its water activity under different saturated environments. The microbial biodegradation of OMW during its storage in 5 evaporation ponds located in Agareb (Sfax-Tunisia) was carried out during the oil-harvesting year held 105 days in 2004. Gravimetric static method using saturated salt solutions was used and OMW as placed at 30 degrees C and under different water activities ranging from 0.11 to 0.90. Eight models were taken from the literature to describe experimental desorption isotherms. During storage, the evolution of physico-chemical parameters including pH, temperature, evaporation, humidity, total phosphorus, chemical oxygen demand (COD), biological oxygen demand (BOD) and phenols and three microbiological flora (aerobic mesophilic bacteria, yeasts and moulds) were considered. At 30 degrees C, when relative humidity increased in the experimented ponds of 69, 84 and 90%, the evaporation speed decreased from 1.24 x 10(-5) to 5 x 10(-6) cm(3) s(-1), from 6 x 10(-5) to 7 x 10(-6) cm(3) s(-1) and from 5 x 10(-6) to 1.1 x 10(-7) cm(3) s(-1) respectively. The desorption isotherm exhibited a sigmoidal curve corresponding to type II, typical of many organic material. The GAB and Peleg models gave the best fit for describing the relationship between the equilibrium moisture content and water activity in OMW (R (2) = 0.998). During the storage period, the analysis showed an increase of all the physico-chemical parameters studied, except phenols and total phosphorus concentrations. The microbiological study showed the predominance of yeasts and moulds and the decrease of bacteria population after 75 days reflecting both effect of recalcitrant compounds and the water activity on microbial growth.