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Essential oil (EO) of Laurus nobilis, from Tunisian, France and Austrian were screened for their chemical composition, antioxidant and antimicrobial activities and compared. GC-MS analysis showed that leaves of Tunisian L. nobilis had camphor (34.43%), 1,8-cineole (20.21%) and α-terpineol (7%) as major components. France and Austrian EOs had a high content of 1,8-cineole (45.8% and 43.4%, respectively) followed by bornyl acetate (13.8% and 17.7% respectively) and methyl eugenol (7.7% and 10.9% respectively). Antioxidant potential was measured by ABTS and DPPH tests. Tunisian L. nobilis EO showed greater radical scavenging by ABTS activity (IC50=44.8±0.1 mg/L) than the France and Austrian EOs (76.4±3.2 mg/L and 81.4±4.0 mg/L, respectively). However, for DPPH test system, French and Austrian EOs activities were excellent (IC50=176.1±5.1 mg/L and 236.3±2.9 mg/L respectively) then Tunisian L. nobilis EO (IC50=2859.7±99.0 mg/L). A good Antimicrobial activity was observed on the yeasts and fungi for all EOs. Tunisian laurel EO show a better antibacterial activity against gram-negative bacteria (Klebsiella pneumonial, E. coli and Salmonella enterica CMI: 0.004 mg/ml) than gram-positive ones (Bacilus subtilis, Staphylococcus aureus and Listeria monocytogenes CMI: 0.01 mg/ml). A significant antifungal activity of Tunisian EO was also observed against fungi and yeasts species (CMI: 0.004 mg/ml). France essential oil shows better activities against all organisms tested wail Austrian oil activity is more important against yeasts species tested and Mucor ramannianus (fungi). Chemical composition, antimicrobial and antioxidant activity of Tunisian L. nobilis essential oil, were different from that of France and Austrian and it give the opportunity for its uses in new pharmaceuticals and natural therapies of infectious diseases.

The petroleum sludge composting in a bioreactor with green wastes was made in interest to assess the organic matter biotransformation, the total petroleum hydrocarbons biodegradation and the maturity of the final product. During the composting process the C/N and NH4+/NO3− ratios decreased to reach values of maturity for the second test B. Otherwise, the decomposition rate was found to be high for test B, as the maximal temperature reached 52 °C for test B while it was only 40 °C for test A. The humification process was monitored by the increase of the humic substance and the HA/FA ratio, and by the FTIR which demonstrated the decrease of the aliphatic group bands and the persistence of the aromatic group bands. The three analysis methods revealed a similarity in the assessment of total petroleum hydrocarbons removal, which reached a rate of 94%. The composting process had also led to a reduction of the phytotoxicity by increasing the germination index.

The purpose of this study is to assess the maturity and stability of the lagooning sludge (LS) with green waste (GW) during semi-industrial composting. Physico-chemical parameters, spectroscopy analysis and germination test were monitored. After 105 days, the decomposition rate of about 33.6; 39 and 36%, a stabilization of pH around neutrality; NH4+/NO3− ratio decreased to 0.922, 0.974 and 1.038; the C/N ratio decreased to 9.47, 9.24 and 10.10 respectively for mixture 1 (M1), mixture 2 (M2) and mixture 3 (M3). This reduction is explained by the bio-oxidation of organic matter. The intense microbial activity was characterized by the rise of temperature (between 40 and 50 °C) during the first weeks. Humification process was characterized by the increase of humification index (HI), which was approximately 62.1%; 73.6%; 61.5% respectively for M1, M2 and M3. The progress of humification process was determined by the decrease of aliphatic absorbance bands and the increase of aromatic absorbance bands. The increase in germination index (GI) (> 50%) for the four seeds: cress, alfalfa, turnip and radish, at the end of composting indicated that the composts are phytotoxic-free and are rich in stable organic compounds and nutrients. The results show that all three composts are mature, which opens the way for their application in agriculture without risk for the soil–plant system.

Saccharothrix algeriensis NRRL B-24137 is an actinobacterium isolated from Algerian Saharan soil. It produces bioactive compounds belonging to the dithiolopyrrolone class of antibiotics, which are characterized by the possession of a unique pyrrolinonodithiole nucleus. Dithiolopyrrolones are known for their strong antibacterial and antifungal activities. This class of antibiotics generated great interest after the discovery of their anticancer properties. In this study, an antibiotic named PR11, produced after a long bacterial fermentation (11 days) in sorbic acid-containing culture broth, was characterized as a new dithiolopyrrolone derivative. After HPLC analysis and purification, the chemical structure of this antibiotic was determined by 1 H- and 13 C-nuclear magnetic resonance, mass and UV-visible data. PR11 was thus characterized as an iso-hexanoyl-pyrrothine, a novel dithiolopyrrolone derivative. The minimum inhibitory concentrations of the new induced antibiotic were determined against several pathogenic micro-organisms. A moderate to strong activity was noted against all Gram-positive bacteria, filamentous fungi and yeasts tested. SIGNIFICANCE AND IMPACT OF THE STUDY: Given the strong activities of dithiolopyrrolones against diverse prokaryotic and eukaryotic micro-organisms including potent selective-anticancer activity, the discovery of new-related derivatives draw continuous attention for therapeutic research. Depending on nature and concentration of added precursor, Saccharothrix algeriensis NRRL B-24137 produce several dithiolopyrrolone coumpounds. In this study, sorbic acid addition combined to long fermentation duration was shown to induce the biosynthesis of a novel dithiolopyrrolone derivative. After purification and full spectroscopic and spectrometric study, the compound was characterized as iso-hexanoyl-pyrrothine. In the future investigation for novel dithiolopyrrolone discovery, fermentation duration should be regarded as a key parameter as well.

Dithiolopyrrolone antibiotics, produced by several micro-organisms, are known for their strong antimicrobial and antitumor activities. Among of this micro-organisms, Saccharothrix algeriensis NRRL B-24137, a rare actinobacterium, has the ability to produce several dithiolopyrrolones derivatives depending on precursors added in the culture medium. After 10 days of strain fermentation on semi-synthetic medium supplemented with cinnamic acid and HPLC purification, biosynthesis of benzoyl-pyrrothine dithiolopyrrolone was evidenced through complete spectroscopic (UV-visible and 1H and 13C NMR) and spectrometric (electron impact mass spectrum) analyses. The pure molecule showed appreciable minimum inhibitory concentration values against several Gram-positive bacteria and filamentous fungi. SIGNIFICANCE AND IMPACT OF THE STUDY: Dithiolopyrrolone antibiotics, known for their strong antimicrobial activities, gained greater interest after the discovery of their antitumor properties. Depending on precursors added, Saccharothrix algeriensis NRRL B-24137 has the ability to produce several dithiolopyrrolones derivatives. Since biological activities of dithiolopyrrolones are related to their variable structure, discover of new natural analogues to be therapeutically explored remains a significant framework of research. In this study, a new dithiolopyrrolone derivative was purified from the fermentation broth of S. algeriensis NRRL B-24137. This new antibiotic, characterized as benzoyl-pyrrothine dithiolopyrrolone, was induced by adding cinnamic acid, as precursor, to a semi-synthetic medium.

In this investigation, petroleum sludge landfilling was carried out in order to assess the biodegradation degree and the final product quality. The microbial analysis showed a good microorganism proliferation which reinforces the biodegradation process. The total mesophilic and thermophilic microflora evaluated symmetrically as they increased at the intermediate stage and decreased at the final. The C/N and NH4+/NO3-ratios decreased while the polymerization degree increased at the end of the landfilling process. The total polyphenols and total petroleum C6 to C22hydrocarbons were removed by 71.6% and 73% respectively, and that affected the reduction of the phytotoxicity in a positive way. All these changes are in agreement with the efficiency of the biotransformation process and showed that petroleum sludge and filling reduced the toxic organic compounds and led to a stable final product.

During screening for potentially antimicrobial actinobacteria, a highly antagonistic strain, designated WAB9, was isolated from a Saharan soil of Algeria. A polyphasic approach characterized the strain taxonomically as a member of the genus Streptomyces. The strain WAB9 exhibited a broad spectrum of antimicrobial activity toward various multidrug-resistant micro-organisms. A PCR-based assay of genomic potential for producing bioactive metabolites revealed the presence of PKS-II gene. After 6 days of strain fermentation, one bioactive compound was extracted from the remaining aqueous phase and then purified by HPLC. The chemical structure of the compound was determined by spectroscopic (UV-visible, and (1)H and (13)C NMR) and spectrometric analysis. The compound was identified to be 2-amino-N-(2-amino-3-phenylpropanoyl)-N-hydroxy-3-phenylpropanamide, a novel hydroxamic acid-containing molecule. The pure molecule showed appreciable minimum inhibitory concentration values against a selection of drug-resistant bacteria, filamentous fungi and yeasts. Significance and impact of the study: This study presents the isolation of a Streptomyces strain, named WAB9, from a Saharan soil in Algeria. This strain was found to produce a new hydroxamic acid-containing molecule with interesting antimicrobial activities towards various multidrug-resistant micro-organisms. Although hydroxamic acid-containing molecules are known to exhibit low toxicities in general, only real evaluations of the toxicity levels could decide on the applications for which this new molecule is potentially most appropriate. Thus, this article provides a new framework of research.

The effects of culture media (YES, CYA, SNM), temperature (10, 15, 20, 25, 30 and 37°C), incubation time (5, 10, 15 and 20 days) and their interactions on radial growth and OTA production by six A. tubingensis and two A. niger were studied. Growth rate increased as temperature increased, regardless of the culture media and isolates. Total inhibition of fungal growth of all strains on the three culture media was observed at 10°C. Mean growth rates calcu-lated for A. niger (2 isolates) and A. tubingensis (6 isolates) revealed that optimal temperatures for growth were observed at 30-37°C on CYA and YES media with maximal growth rates of 5.7 and 5.4 mm/day for A. niger and A. tubingensis cultivated in YES medium, respectively. Growth on SNM medium is generally slow, regardless the temperature and strain used. For both spe-cies, maximum means growth rates were determined at 37°C (3.7-3.8 mm/day). For OTA production, an incubation period of 5 days is sufficient to de-tect the OTA. Levels of this toxin are dependent on temperature, incubation time and strain. In the A. tubingensis specie, maximum of 0.710 μg/g of OTA was obtained after 10 days of incubation at 25°C on SNM medium while for A. niger , the maximum (0.36 μg/g) was obtained on CYA medium. The opti-mum conditions for OTA production are 25 °C/10 days. Analysis of variance showed that the effects of single factors (culture medium, temperature, in-cubation time and strain) and their interactions on growth and OTA produc-tion were highly significant (P=0.000).

Simultaneous in vivo measurements of prompt fluorescence (PF), delayed fluorescence (DF) and modulated reflection (MR) at 820-nm were used to assess effect of short desiccation period (4.5 h) on Parmelina tiliacea lichen. The two performance indexes (PIABS and PItotal) as a measure of an overall photosynthetic thalli performance showed a negative effect of desiccation treatment on photosynthetic activity. The maximal intensity of PF and DF recorded during desiccation treatment decreased and at 4.5 h desiccation time Parmelina tiliacea thalli loss their variable fluorescence and DF amplitude. This loss of variable fluorescence was due to an increase in inactive reaction centers and a limitation of electron donation on the donor side of photosystem II (PSII) that caused a down-regulation of electron transport chain at the PSII level. However, the efficiency with which an electron can move from the reduced intersystem electron acceptors to the PSI end electron acceptors was less affect by desiccation treatment. In respect to MR change, re-reduction kinetics of the primary electron donors of photosystems I (P700) and plastocyanin (PC) in desiccated thalli seem to be faster, their amplitudes gradually decrease and a disconnection between the two photosystem (PSII and PSI) were observed. These responses allow to Parmelina tiliacea lichen a photoprotection mechanism from the excess light excitation.

A halophilic actinomycete strain, designated H55T, was isolated from Saharan soil sampled in the Mzab region (Ghardaïa, southern Algeria) and was characterized in a taxonomic study using a polyphasic approach. The cell wall was determined to contain meso-diaminopimelic acid and the characteristic whole-cell sugars were arabinose and galactose. The predominant menaquinones were found to be MK-10(H4) and MK-9(H4). The predominant cellular fatty acids were determined to be anteiso-C17 : 0, iso-C16 : 0 and iso-C15 : 0. The diagnostic phospholipid detected was phosphatidylcholine. The morphological and chemotaxonomic characteristics of the strain were consistent with those of members of the genus Actinopolyspora , and 16S rRNA gene sequence analysis confirmed that strain H55T was a member of this genus. DNA–DNA hybridization values between strain H55T and the type strains of the nearest species of the genus Actinopolyspora , Actinopolyspora erythraea and A. alba , were clearly below the 70 % threshold. The genotypic and phenotypic data showed that the organism represents a novel species of the genus Actinopolyspora , for which the name Actinopolyspora mzabensis sp. nov. is proposed, with the type strain H55T ( = DSM 45460T = CCUG 62965T).

International audience; Actinomycete strain SA198, isolated from a Saharan soil sample of Algeria, exhibited antimicrobial activity against Gram-positive and Gram-negative bacteria, and phytopathogenic and toxinogenic fungi. The morphological and chemotaxonomic characteristics of the strain were consistent with those of the genus Saccharothrix. Analysis of the 16S rRNA gene sequence of strain SA198 showed a similarity level ranging between 97.2 and 98.8% within Saccharothrix species, S. australiensis being the most closely related. Two new active products were isolated by reverse HPLC using a C18 column. The ultraviolet-visible (UV-VIS), infrared (IR), mass, and 1Hand 14C nuclear magnetic resonance (NMR) spectra showed that these products were new bioactive compounds. The minimum inhibitory concentrations of these antibiotics showed a strong activity against fungi and moderate activities against Gram-positive and Gram-negative bacteria.

An actinomycete, designated SA181T, was isolated from Saharan soil in the Hoggar region (south Algeria) and was characterized taxonomically by using a polyphasic approach. The morphological and chemotaxonomic characteristics of the isolate were consistent with the genus Saccharothrix , and 16S rRNA gene sequence analysis confirmed that strain SA181T was a novel member of the genus Saccharothrix . DNA–DNA hybridization values between strain SA181T and its closest phylogenetic neighbours, the type strains of Saccharothrix longispora , Saccharothrix texasensis and Saccharothrix xinjiangensis , were clearly below the 70 % threshold. The genotypic and phenotypic data showed that the isolate represents a novel species of the genus Saccharothrix , for which the name Saccharothrix hoggarensis sp. nov. is proposed, with the type strain SA181T ( = DSM 45457T = CCUG 60214T).

Ochratoxine A (OTA), a potential human carcinogen is produced by several species of Aspergillus and Penicillium, including Aspergillus westerdijkiae. In this study a putative polyketide synthase gene aolc35-12 has been partially cloned from A. westerdijkiae. The predicted amino acid sequence of the 3.22 kb clone was found to have a high degree of similarity to other previously identified polyketide synthase genes from various OTA-producing fungi including Aspergillus ochraceus, Aspergillus niger, Aspergillus carbonarius and Penicillium nordicum. The aolc35-12 gene was disrupted and inactivated by insertion of Escherichia coli hygromycin B phosphotransferase gene, which resulted in an OTA negative mutant aoΔlc35-12. Genetic complementation confirmed aolc35-12 as OTA-polyketide synthase gene. Furthermore, study of the differential expression of aolc35-12 and a previously identified OTA-polyketide synthase gene, i.e. aoks1, in the wild-type A. westerdijkiae and aoΔlc35-12 mutant revealed that aolc35-12 could code for a certain polyketide compound complementary for the expression of aoks1 and hence for the activation of OTA biosynthesis system in A. westerdijkiae.

A new halophilic actinomycete strain, designated H16, has been isolated from a hypersaline Saharan soil in Ouargla province (southern Algeria) and characterized taxonomically using a polyphasic approach. The strain grew at 18–50°C, pH 5–9, and 7–32% (w/v) NaCl. It produced abundant aerial mycelia, which formed long chains of rod-shaped spores at maturity, and fragmented substrate mycelia. The strain contained chemotaxonomic markers that were diagnostic for the genus Actinopolyspora, such as meso-diaminopimelic acid, arabinose, and galactose, phosphatidylcholine as a diagnostic phospholipid, and predominant menaquinones MK-9(H4) and MK-10(H4). The predominant fatty acids were anteiso- and iso-C17:0, iso-C15:0, and 9-methyl-C16:0. Phylogenetic analysis, based on 16S rDNA gene sequences, confirmed that strain H16 is a member of the genus Actinopolyspora and most closely related to A. mortivallis (98.7% identity). DNA–DNA hybridization confirmed that it belonged to A. mortivallis. This strain showed high activity against Klebsiella pneumoniae and optimally secreted antibiotics during the mid-stationary growth phase in liquid culture. The antibiotics were extracted with n-butanol and separated on silica gel plates by ethanol–ammonia–water (mobile phase). The results of bioautography revealed the presence of five antibiotics. The chemical revelations showed that these antibiotics were glycosylated polycyclic aromatic compounds containing amine groups and hydroxamic acids. The UV-visible and mass spectra of the most active antibiotics were determined.

The diversity of a population of 52 halophilic actinomycetes was evaluated by a polyphasic approach, which showed the presence of members of the Actinopolyspora , Nocardiopsis , Saccharomonospora , Streptomonospora , and Saccharopolyspora genera. One strain was considered to be a new member of the last genus, and several other strains seemed to be new species. Furthermore, 50% of strains were active against a broad range of indicators and contained genes encoding polyketide synthetases and nonribosomal peptide synthetases.

A moderately halophilic actinomycete strain designated AH97 was isolated from a saline Saharan soil, and selected for its antimicrobial activities against bacteria and fungi. The AH97 strain was identified by morphological, chemotaxonomic and phylogenetic analyses to the genus Actinoalloteichus. Analysis of the 16S rDNA sequence of strain AH97 showed a similarity level ranging between 95.8% and 98.4% within Actinoalloteichus species, with A. hymeniacidonis the most closely related. The comparison of the physiological characteristics of AH97 with those of known species of Actinoalloteichus showed significant differences. Strain AH97 showed an antibacterial and antifungal activity against broad spectrum of microorganisms known to be human and plant pathogens. The bioactive compounds were extracted from the filtrate culture with n-butanol and purified using thin layer chromatography and high pressure liquid chromatography procedures. Two active products were isolated, one hydrophilic fraction (F1) and another hydrophobic (F2). Ultraviolet-visible, infrared, mass and (1)H and (13)C nuclear magnetic resonance spectroscopy studies suggested that these molecules were the dioctyl phthalate (F2) and an aminoglycosidic compound (F1).

Dithiolopyrrolone antibiotics, produced by several microorganisms, are known for their strong antimicrobial activities. This class of antibiotics generated new interest after the discovery of their anticancer and antitumor properties. In this study, four new antibiotics were purified from the fermentation broth of Saccharothrix algeriensis NRRL B-24137 and characterized as dithiolopyrrolone derivatives. These new dithiolopyrrolone antibiotics were induced by adding sorbic acid, as precursor, at a concentration of 5 mM to the semi-synthetic medium. The analysis of the induced antibiotics was carried out by HPLC. The maximal production of the antibiotics PR2, PR8, PR9 and PR10 was 0.08±0.04, 0.21±0.04, 0.13±0.03 and 0.09±0.00 mg L(-1) , respectively, obtained after 8 days of fermentation. The chemical structures of these antibiotics were determined by (1) H- and (13) C-nuclear magnetic resonance, mass and UV-visible data. The four new dithiolopyrrolone antibiotics - PR2, PR8, PR9 and PR10 - were characterized, respectively, as crotonyl-pyrrothine, sorbyl-pyrrothine, 2-hexonyl-pyrrothine and 2-methyl-3-pentenyl-pyrrothine. The minimum inhibitory concentrations of the new induced antibiotics were determined.

Two novel halophilic actinomycetes, strains AH35 and AH10, isolated from saline soils obtained from the Algerian Sahara, were subjected to polyphasic taxonomic characterization. The strains showed good growth in medium containing 7, 10 and 15% NaCl with growth at temperatures of 28–42°C. The organism produced light grey-green aerial mycelium with reddish-brown or yellow-brown diffusible pigment. Micromorphology and chemotaxonomic characteristics were consistent with its assignment to the genus Saccharomonospora. Analysis of 16S rDNA gene sequences showed that AH35 and AH10 formed a robust clade with “Actinopolyspora iraqiensis”, Saccharomonospora halophila and Saccharomonospora paurometabolica. However, a number of phenotypic characteristics readily distinguished strains AH35 and AH10 from these three species. The two strains showed antibacterial activity against Gram-negative bacteria such as Klebsiella pneumoniae, Salmonella enterica, Pseudomonas syringae and Agrobacterium tumefaciens.

The primary goal of this research was to assess the biotransformation of zearalenone (ZEA), an oestrogenic mycotoxin, into ZEA-sulfate by some Aspergillus section Nigri isolates. A. niger isolates were shown to be able to convert ZEA after 72 h of incubation at different ZEA concentrations (5 to 150 µg/ml). The product formula corresponding to ZEA-sulfate was determined by accurate mass measurement. This conjugate was shown to be less oestrogenic than ZEA using a MCF-7 proliferation assay. This study demonstrated that A. niger has the ability to transform ZEA over a broad range of ZEA concentrations (5 to 150 µg/ml) and that sulfonation could lead to a less toxic compound. These results are significant as little is known about the ability of fungi, and especially A. niger, to detoxify ZEA by sulfonation. This study could lead to a way of detoxifying feed naturally contaminated with mycotoxins.

Zearalenone (ZEA) is an oestrogenic mycotoxin produced by Fusarium spp. on a wide variety of grains. The presence of ZEA impairs farm animal health. In this study, the ability to remove ZEA by conidia of Aspergilli was evaluated in vitro. This mycotoxin was successfully eliminated in vitro by living and heat-treated conidia just after conidial inoculation, especially by Aspergillus japonicus Mu541 (IMI389204) conidia. Consequently, components involved in adsorption are not affected by heat-treatment. The influence of pH on ZEA removal was also determined. Results have shown that ZEA removal was 53% at pH 2 while 2% removal was observed at pH 11. Then, the adsorption capacity was evaluated in vitro in porcine gastrointestinal tract conditions. We observed that there was better adsorption in acidic stomach conditions. Finally, the phenomenon has been modelled using Hill's equation. This model was very suitable and confirmed better adsorption at acidic pH. Aspergilli section Nigri conidia were able to adsorb ZEA instantaneously especially in acidic conditions as in the stomach. The use of heat-treated conidia could be a very interesting means of decreasing ZEA levels in animal feed. It must now be assessed directly on naturally contaminated feed and on animals.

International audience; Aspergillus westerdijkiae is the main producer of several biologically active polyketide metabolites including isoasperlactone and asperlactone. A 5298 bp polyketide synthase gene ‘‘aomsas” has been cloned in Aspergillus westerdijkiae by using gene walking approach and RACE-PCR. The predicted amino acid sequence of aomsas shows an identity of 40–56% with different methylsalicylic acid synthase genes found in Byssochlamys nivea, P. patulum, A. terreus and Streptomyces viridochromogenes. Based on the reverse transcription PCR and kinetic secondary metabolites production studies, aomsas expression was found to be associated with the biosynthesis of isoasperlactone and asperlactone. Moreover an aomsas knockout mutant ‘‘aoDmsas” of A. westerdijkiae, not only lost the capacity to produce isoasperlactone and asperlactone, but also 6-methylsalicylic acid. The genetically complemented mutant ao+msas restored the biosynthesis of all the missing metabolites. Chemical complementation through the addition of 6-methylsalicylic acid, aspyrone and diepoxide to growing culture of aoDmsas mutant revealed that these compounds play intermediate roles in the biosynthesis of asperlactone and isoasperlactone.

International audience; GC-FID and GC-MS analysis of essential oils of Juniperus phoenicea resulted in the identification of 30 compounds, representing more than 98% of the total composition. α-pinene (55.7% and 80.7%), δ-3-carene (10.7% and 4.5%), and γ-cadinene (2.9% and 5.1%) were the main components, respectively, in leaves and berries essential oil. Extracts of J. phoenicea were obtained by different extraction solvents: methanol, ethanol, ethyl acetate, and dichloromethane and evaluated composition for polyphenols (gallic acid equivalent 52 to 217 g/kg), tannins (catechin equivalent 6.5 to 60.2 g/kg), antocyanins (cyanidin equivalent 84 to 373 mg/kg), and flavonoids (quercetin equivalent 6.4 to 29.3 g/kg). The samples (essential oils and extracts) were subjected to a screening for their antioxidant activity by using DPPH and ABTS assays; antimicrobial activity was tested with 6 bacteria (3 Gram-positive and 3 Gram-negative), 1 yeast, and 2 fungi. The strongest antioxidant activity was obtained by the methanolic extract (IC50= 6.5 ± 0.3 mg/L). Flavonoids are likely to contribute to the antifungal activity against Saccharomyces cerevisiae. Correlations were studied between chemical composition and antioxidant and antimicrobial activities.

International audience; The study was aimed to investigate essential oil chemical composition (gas chromatography/flame ionization detection [GC-FID] and gas chromatography/mass spectrometry [GC-MS]) and antioxidant (1,1-diphenyl-2-picrylhydrazyl free radical (DPPH) and 2,2’-azinobis--ethylbenzothiazoline-6-sulphonate [ABTS] assays) and antimicrobial (Gram-positive and Gram-negative bacteria, fungi, and yeast) activities of essential oils extracted from leaves of Mentha longifolia L. and Mentha viridis. GC-MS analysis revealed that M. longifolia was constituted by pulegone (54.41%) as a major component followed by isomenthone (12.02%), 1,8-cineole (7.41%), borneol (6.85%),and piperitenone oxide (3.19%). M. viridis was rich in carvone (50.47%), 1,8-cineole (9.14%), and limonene (4.87%).The antioxidant activity by ABTS assay showed IC50 values of 476.3 ± 11.7 and 195.1 ± 4.2 mg/L for M. longifolia and M. viridis, respectively, the DPPH assays have resulted in a moderate IC50 (>8000 mg/L and 3476.3 ± 133 mg/L for M. longifolia and M. viridis, respectively). Antimicrobial activity showed that Listeria monocytogenes and Klebsiella pneumoniae bacteria were more inhibited by the 2 essential oils tested. Escherichia coli was least susceptible. A strong activity was also observed on fungi and yeasts. Carvone, thymol, and piperitone oxide have not been detected in Tunisian M. longifolia. Camphor is reported for the 1st time for M. viridis. Antioxidant and antibacterial activities were correlated to chemical composition.

The natural dithiolopyrrolone antibiotics, produced during fermentation by Saccharothrix algeriensis NRRL B-24137, are known for their potent antibacterial, antifungal, and anticancer activities. During dithiolopyrrolone biosynthesis, the activated organic acid (acyl-CoA) is incorporated into the pyrrothine nucleus. We investigated the use of precursor-directed biosynthesis (PDB) to create new dithiolopyrrolone analogs by substitution of several organic acids in place of the usual organic acid. No other study on dithiolopyrrolone production by precursor-directed biosynthesis has been reported. The production of four novel dithiolopyrrolones in the presence of benzoic, valeric and cinnamic acids by S. algeriensis was obtained in this study. The mass spectrometry data showed a direct incorporation of added valeric and benzoic acids to form several dithiolopyrrolone analogs. However, addition of all other organic acids did not lead to the production of the corresponding dithiolopyrrolone analogs. We describe here the use of the PDB process for production of four new dithiolopyrrolone analogs, valeryl-pyrrothine, benzoyl-pyrrothine, demethyl-benzoyl-pyrrothine and formoyl-pyrrothine.

Wheat is a basic staple food for very large segments of the population of Algeria. The aim of this study is to analyse ochratoxin A (OTA)-producing mould and OTA-contaminated wheat. To evaluate the mycoflora and the potential for OTA production by Aspergillus strains, a total of 85 samples of wheat destined for human consumption were collected from two regions in Algeria (Tizi Ouzou and Setif) during the following phases: preharvest, storage in silos, and after processing. The mean value counts of fungi ranged from 275 to 1277 CFU g(-1). The dominant genus was Aspergillus, predominantly A. flavus, A. niger and A. versicolor. The other isolated species were A. ochraceus, A. alliaceus, A. carbonarius, A. terreus, A. fumigatus, A. candidus and Aspergillus spp. The occurrence and the levels of the genus Penicillium, Fusarium, Alternaria and Mucor were substantially lower than those of Aspergillus. The storage in silos shows high levels of Aspergillus (66 to 84%), especially A. flavus, but A. niger and other fungi were isolated at relatively low percentages. Equal distribution of the fungal contamination into the bran, flour and semolina fractions was observed from Flour Mill and Semolina Mill. The genus Aspergillus remained present at high levels at several phases of the production process. In addition, the ability to produce OTA by 135 isolates belonging to eleven species of Aspergillus and 23 isolates of Penicillium spp. was analyzed using fluorescent detection-based HPLC. Thus, it was found that 51 isolates (32.3%) were ochratoxigenic. All isolated strains of A. ochraceus (12) and A. alliaceus (6) produced OTA at concentrations ranging from 0.23 to 11.50 microg g(-1). Most of the A. carbonarius strains (80%) were OTA producers (0.01 to 9.35 microg g(-1)), whereas A. terreus (50%), A. niger (28%), A. fumigatus (40%), A. versicolor (18%) and Penicillium spp. (21.7%) were low level producers (0.01 to 0.07 microg g(-1)). The concentration of OTA was determined in 30 samples of wheat. OTA was detected in 12 (40%) of the samples at levels ranging from 0.21 to 41.55 microg kg(-1).

Dans cette étude, l’influence de l’interaction de la température et de l’activité de l’eau sur la croissance fongique et la production de l’ochratoxine A sur un milieu de base café a été évaluée. La croissance optimale des trois souches d’ Aspergillus a été observée dans les mêmes conditions écologiques à savoir, 30 °C pour la température et 0,99 pour l’activité de l’eau. Les croissances journalières maximales sont 11,2, 6,92 et 7,22 mm/jour respectivement pour la souche d’Aspergillus niger, d’Aspergillus carbonarius et d’Aspergillus ochraceus. Cependant, les conditions écologiques de production optimale d'ochratoxine A varient selon la souche toxinogène avec comme facteur limitant, l’activité de l’eau. Cette production d’ochratoxine A est inhibée à la température de 42 °C et à l’activité de l’eau de 0,75. La correspondance établie entre les valeurs d’activité de l’eau testées en laboratoire et celles mesurées sur un lot de cerises mûres mis à sécher au soleil montre que les 5 premiers jours de séchage constituent la phase critique de multiplication fongique et de production de l’ochratoxine A. Ainsi, un séchage artificiel des cerises à une température supérieure à 42 °C évitera non seulement la multiplication fongique mais également la contamination de ce produit par l’ochratoxine A.

The influence of organic acids on growth and dithiolopyrrolone antibiotic production by Saccharothrix algeriensis NRRL B-24137 was studied. The production of dithiolopyrrolones depends upon the nature and concentration of the organic acids in the culture medium. Study of the nature of organic acids showed that the most effective organic acids for thiolutin specific production were maleic, 4-hydroxybenzoic, benzentetracarboxylic, pantothenic, pivalic and pyruvic acids (which yielded almost five-fold over the starting medium) and pimelic acid (more than three-fold). 4-Bromobenzoic acid showed the best production of senecioyl-pyrrothine (59 mg g−1 DCW). Tiglic acid showed the best production of tigloyl-pyrrothine (22 mg g−1 DCW). The highest yield of isobutyryl-pyrrothine (7.6 mg g−1 DCW) was observed in the presence of crotonic acid. Sorbic acid yielded the best production of butanoyl-pyrrothine (26 mg g−1 DCW). Methacrylic, butyric, pyruvic and 4-bromobenzoic acids also exhibited the best production of butanoyl-pyrrothine (27–11-fold). Study of organic acid concentration showed that among the selected organic acids, pimelic acid yielded the highest specific production of thiolutin (91 mg g−1 DCW) at 7.5 mM; and senecioyl-pyrrothine (11 mg g−1 DCW), tigloyl-pyrrothine (9 mg g−1 DCW) and butanoyl-pyrrothine (3.5 mg g−1 DCW) at 5 mM. Pyruvic acid at 1.25 mM enhanced the production of senecioyl-pyrrothine (4.3 mg g−1 DCW). The maximum production of tigloyl-pyrrothine (18.6 mg g−1 DCW) was observed in the presence of tiglic acid at 2.5 mM. Maximum production of isobutyryl-pyrrothine was observed in the presence of 7.5 mM tiglic acid. In addition, methacrylic acid (at 5 mM) and butyric acid (at 2.5 mM) enhanced the production of butanoyl-pyrrothine (26 and 20 times, respectively). The above results can be employed in the optimisation of the culture medium for the production of dithiolopyrrolone in higher quantities.

Ten antibiotic-producing Streptomyces spp. isolated from Moroccan soils were evaluated for their ability to inhibit in vitro Sclerotium rolfsii development. Four isolates having the greatest pathogen inhibitory capabilities were subsequently tested for their ability to inhibit sclerotial germination in sterile soil. This test was carried out by using biomass inoculum, culture filtrate, and spore suspension of the isolates as treatment. Treatment with biomass inoculum and culture filtrate gave the highest inhibition of sclerotia. Biological control tests against Sclerotium rolfsii damping-off of sugar beet seeds showed that the selected Streptomyces isolates reduced significantly the disease severity, the J-2 isolate being the more potent. In addition, treatment with the isolate J-2 resulted in a significant increase (P ≤ 0.05) in seedling development compared to the control. All antagonistic Streptomyces selected here were able to grow in the rhizosphere soil from infected sugar beet culture.

International audience; Deoxynivalenol (DON) is a mycotoxin affecting animals and plants. This toxin synthesized by Fusarium culmorum and Fusarium graminearum is currently believed to play a decisive role in the fungal phytopathogenesis as a virulence factor. Using cultured cells of Nicotiana tabacum BY2, we showed that DON-induced programmed cell death (PCD) could require transcription and translation processes, in contrast to what was observed in animal cells. DON could induce different cross-linked pathways involving (i) reactive oxygen species (ROS) generation linked, at least partly, to a mitochondrial dysfunction and a transcriptional down-regulation of the alternative oxidase (Aox1) gene and (ii) regulation of ion channel activities participating in cell shrinkage, to achieve PCD.

This paper reports the results of an extensive survey on the occurrence of filamentous fungi isolated from wine-grapes in Lebanon and to test their ability to produce ochratoxin A (OTA) and aflatoxin B1 (AFB1) on CYA culture medium, in order to assess their potential for producing these mycotoxins on grapes. From the 470 grapes samples taken during season 2004, 550 fungi strains were isolated with 490 belonging to Aspergillus spp. and 60 belonging to Penicillium spp. All these isolated fungi starins were tested for their ability to produce OTA and AFB1. Aspergillus carbonarius shows that it is the only species able to produce OTA with a production percentage reaching 100% and a maximum concentration of 52.8 microg/g of Czapek yeast extract agar (CYA). In its turn, Aspergillus flavus was considered as the only AFB1-producing species with production percentage of 45.3% and a maximum concentration reaching 40 microg/g CYA. A total of 47 handmade musts produced from the collected grapes were also analyzed in order to correlate the presence of OTA in must and the occurrence of filamentous fungi on grapes; 57.4% were contaminated with OTA at low level with concentrations ranging between 0.011 and 0.221 microg OTA L(-1). The analysis of these must samples was not performed with regard to AFB1. Seventy samples of finish red wine were also assayed for OTA content. The results showed that 42 of the tested samples (60%) were found to be positive for OTA with low levels (0.012-0.126 microg OTA L(-1)).

The effect of the frequency of the mixing of coffee cherries put out for sun drying on the kinetics of the drying, fungal growth and kinetics of ochratoxin A production was evaluated. The results showed that the more coffee cherries were mixed, the quicker they dried. This rapidity of drying led to a reduction of fungal development. Indeed, coffee cherries mixed eight and ten times a day, dried quickly and were free inside of fungi. However, infection by fungi gives little indication of ochratoxin A production in coffee cherries. Indeed, although coffee cherries mixed twice a day were more contaminated by fungi, the analysis of ochratoxin A content showed they were free of this mycotoxin. The coffee cherries that were more contaminated by ochratoxin A were those mixed four times a day (containing 0.35-5.46 microg kg(-1) ochratoxin A). Ochratoxin A contamination was essentially due to the presence of Aspergillus species capable of producing ochratoxin A inside the coffee cherries.

Twenty-five soil samples were collected in the Algerian Sahara and analyzed to isolate rare actinomycetes. Eighty-six isolates with the same Nocardiopsis or Saccharothrix morphology were isolated on humic-vitamin B agar medium using dilution techniques and several antibiotics as selective agents. Certain of these antibiotics seemed to be very selective for some phenotypes. Morphological and chemotaxonomic characteristics led to identifying 54 isolates belonging to the Nocardiopsis genus and 32 isolates belonging to the Saccharothrix genus. An assessment of the antimicrobial properties of the isolates showed activities against Gram-positive bacteria, fungi and yeasts. Saccharothrix isolates possessed better antifungal activity than Nocardiopsis. One of them, labeled SA 103, was therefore selected for identification of its antifungal antibiotic activities. Production of overall antifungal and antibacterial activities was checked on the complex medium ISP2 and a synthetic medium (SM) that contains glucose or starch as carbon source, and ammonium or nitrate as nitrogen source. The SM medium containing ammonium sulfate (0.2%), supplemented with starch (0.5%) and yeast extract (0.3%), was retained for production of antibiotics. Active substances were purified by a G25-80 Sephadex column and reverse phase HPLC. Two pure substances were obtained and named ZA01 and ZA02; they were characterized on the basis of combined data resulting from chemical tests, UV visibile and IR spectra and mass spectrometry. The two antibiotics were found to be related and were partially characterized as nucleotidic or nucleosidic antibiotics. Their structures consisted of a chain of three sugar units linked to an aromatic base containing a phosphate residue.

Fungi contaminating foods and feeds may produce many mycotoxins including ochratoxin A (OTA). Early and rapid detection of potential OTA producing fungi is important to reduce the negative impacts of OTA. In this study, two PCR specific primer pairs, AoLC35-12L/AoLC35-12R and AoOTAL/AoOTAR, were designed from a DNA sequence of a polyketide synthase gene in Aspergillus ochraceus NRRL 3174. On 14 different fungi tested by PCR, AoLC35-12L/AoLC35-12R amplified a unique band from either OTA or citrinin producers while AoOTAL/AoOTAR amplified one PCR product only from A. ochraceus. So these primers could be used to detect both OTA and citrinin producing fungi (AoLC35-12L/AoLC35-12R) or only A. ochraceus (AoOTAL/AoOTAR) from foodstuffs using PCR method.

To provide an efficient technique for monitoring the off-flavoured fungal compound geosmin.Geosmin-associated gpe1 gene of Penicillium expansum displayed ≥99% similarity to cytochrome P450 gene of geosmin-producing P. restrictum, but ≤40% similarities to geosmin biosynthesis, non-cytochromic gene of Streptomyces avermitilis and cytochrome P450 genes of non-geosmin-producing Neotyphodium lolii, Phoma betae and P. paxilli. Serial 10-fold dilutions of P. expansum's DNA was subjected to a previously reported qPCR assay (Atoui et al. 2007), utilizing gpe1 specific primer pair 'SNgpe1F/SNgpe1R'. A linear relationship between DNA quantity and Cycle Threshold (Ct ), with strong correlative coefficient, was observed. Using the available physico-chemical method, geosmin was quantified in 188 grape samples. Penicillium spp's DNA was quantified in these samples, utilizing the developed qPCR assay. A strong positive correlation (R(2) = 0·97) between Penicillium's DNA and geosmin concentration was observed. Furthermore,50 ng μl(-1) Penicillium's DNA corresponds to geosmin level below the permitted intensity limit i.e. 4, for 'Flavour Profile Analysis'.Penicillium spp., genomic DNA level can provide an efficient way to quantify geosmin.This particular qPCR technique can be utilized in numerous food industries, for the timely detection and monitoring of geosmin contamination.

Plantaricin C19, an anti-Listeria bacteriocin, was successfully purified by adsorption to and release from producing cells at low pH combined with reverse phase high-performance liquid chromatography (HPLC). The purification resulted in a 900-fold increase in specific activity with a yield of 15% of the original activity. Mass spectrometry analysis gave a molecular weight of 3845.3. Protein microsequencing identified 36 amino acids. Plantaricin C19 is rich in both hydrophobic and basic amino acids in good accordance with its basic and hydrophobic character. Comparison of the amino acid sequence of plantaricin C19, with the sequence of some other anti-Listeria bacteriocins produced with lactic acid bacteria, revealed that plantaricin C19 has in its N-terminal region the consensus sequence--YYGNGL--(uniquely with Valine instead of Leucine as found in all other bacteriocins), identifying plantaricin C19 as a pediocin-like bacteriocin. Plantaricin C19 exerted a bacteriostatic action on sensitive cells of Listeria grayi IP 6818 in BHI broth. No loss of intracellular K+, Mg2+ or UV-absorbing materials was observed. Adsorption of plantaricin C19 on L. grayi CIP 6818 decreased in the presence of salts.

International audience; The dithiolopyrrolone class of antibiotics has been known to display bacteriostatic activity against both Gram-positive and Gram-negative bacteria and exert other biological activities. Acyltransferase activities are proposed to be responsible for the structural diversity of dithiolopyrrolones produced by Saccharothrix algeriensis NRRL B-24137. Moreover, two activities, pyrrothine N-acetyltransferase and pyrrothine N-benzoyltransferase, are reported to catalyze the formation, respectively, to thiolutin and benzoyl-pyrrothine (BEP) in this bacterium. In this study, two genes encoding two putative acyltransferases were identified in S. algeriensis. The first one, actA, was identified by bioinformatic analysis and by analogy to an acetyltransferase, hlmA, identified in holomycin biosynthetic gene cluster in Streptomyces clavuligerus. The second was identified by purification of both enzymes from the bacterial biomass which provided a semipurified extract. The microsequencing of tryptic peptides from the final protein preparation yielded sequences of eight different fragments, two of them encoded by one gene, actB, in S. algeriensis genome bank. The alignment of actB against the GenBank database revealed significant homology to acyltransferase family. Differential expression of these genes, actA and actB, was then investigated in three different media: (i) semisynthetic medium (SSM), which promotes the production of thiolutin; (ii) SSM supplemented by 1.25 mM benzoic acid (SSM + BA), which promotes the production of both thiolutin and BEP; and (iii) tryptic soy broth (TSB) in which no dithiolopyrrolone derivatives were detected.

Carnobacterium piscicola CP5, isolated from a French mold-ripened soft cheese, produced a bacteriocin activity named carnocin CP5, which inhibited Carnobacterium, Enterococcus and Listeria spp. strains, and among the Lactobacillus spp. only Lactobacillus delbrueckii spp. [24]. The activity was purified by ammonium sulfate precipitation, anion exchange, and hydrophobic interaction chromatography followed by reverse-phase high-performance liquid chromatography (RP-HPLC). This latter step separated two peaks with anti-listerial activity (CP51 and CP52). Carnocin CP51 was partially sequenced, and the N-terminal part revealed the presence of the "pediocin-like consensus" sequence-Tyr-Gly-Asn-Gly-Val-. Then, a degenerated 24-mer oligonucleotide probe was constructed from the N-terminal sequence and used to detect the structural gene. It was localized on a plasmid of about 40 kb. Cloning of restriction fragments of this one, followed by DNA sequencing, revealed the presence of the second anti-Listeria bacteriocin gene (CP52). By comparing sequences in data banks and confirming results with PCR reactions, carnocin CP51 shared homologies with carnobacteriocin BM1, and carnocin CP52 was similar to carnobacteriocin B2, both produced by C. piscicola LV17 [2]. However, carnobacteriocin A from C. piscicola LV17 gene was lacking in C. piscicola CP5, and the two microorganisms have been isolated from different ecological environments: C. piscicola CP5 and C. piscicola LV17 were isolated from soft cheese and vacuum-packed meat respectively. This fact could allow different application perspectives for C. piscicola CP5.

Brevicin 27, a bacteriocin produced by Lactobacillus brevis SB27, is inhibitory mainly against closely related Lactobacillus brevis and Lactobacillus buchneri strains. It was purified from the culture supernatant by a four-step purification procedure including ammonium sulfate precipitation, cation exchange, hydrophobic interaction, and reverse-phase, high performance liquid chromatographies. The purified bacteriocin was subjected to mass spectrometry, amino acid composition analysis, and sequencing by Edman degradation. It was shown to be an about 5200-Da basic protein containing a high proportion of lysine and of hydrophobic amino acids. The partial N-terminal amino acid sequence (25 residues) was unique when compared with the Protein Data Bank (PDB), Swiss Prot, and Protein Information Resource (PIR) data banks and to the translated Gen Bank.

One promising strain Actinomadura sp. AC170, isolated from Algerian Saharan soil, with strong antifungal activity against pathogenic and toxinogenic fungi, was selected for further studies. The 16S rRNA results showed a distinct phylogenetic lineage from the other species within the Actinomadura genus. The production of antibiotic substances was investigated using GYEA solid medium. The butanolic extract contained four bioactive spots detected on thin layer chromatography plates. Among these antibiotics, a complex called 170A, which showed the more interesting antifungal activity, was selected and purified by reverse-phase HPLC. This complex is composed of four compounds. Ultraviolet-visible, infrared, mass and H nuclear magnetic resonance spectroscopy studies showed that these molecules contain an aromatic ring substituted by aliphatic chains. These compounds differ from the known antibiotics produced by Actinomadura species. Key words: Actinomadura, taxonomy, antifungal compounds, Saharan soil.

Neroli essential oil is extracted from the fragrant blossoms of the bitter orange tree. It is one of the most widely used floral oils in perfumery. In this study chemical composition and in vitro antimicrobial and antioxidant activities of neroli oil are investigated. The essential oil of fresh Citrus aurantium L. Flowers (Neroli oil) cultivated in North East of Tunisia (Nabeul) were analyzed by GC-FID and GC-MS. About 33 compounds were identified, representing 99% of the total oil. Limonene (27.5%) was the main component followed by (E)-nerolidol (17.5%), alpha-terpineol (14%), alpha-terpinyl acetate (11.7%) and (E, E)-farnesol (8%). Antimicrobial activity was determined by Agar-well-diffusion method against 6 bacteria (3 Gram-positive and 3 Gram-negative), 2 yeasts and 3 fungi. Neroli oil exhibited a marked antibacterial activity especially against Pseudomonas aeruginosa. Moreover, Neroli oil exhibited a very strong antifungal activity compared with the standard antibiotic (Nystatin) as evidenced by their inhibition zones. Antioxidant activity determined by ABTS assay showed IC50 values of 672 mg L(-1). Finally, this study may be considered as the first report on the biological properties of this essential oil. The results of this study have provided a starting point for the investigations to exploit new natural substances present in the essential oil of C. aurantium L. flowers.

Background and aim Saccharothrix algeriensis NRRL B-24137, isolated from a Saharan soil, has been described as a potential biocontrol agent against Botrytis cinerea and other phytopathogens. However, the plant protection mechanisms involved still need to be described. The aim of this study was to determine this protection phenomenon as well as parts of the mechanisms involved, using Arabidopsis thaliana seedlings and B. cinerea. Methods The bacterial colonization process was evaluated on A. thaliana seedlings using fluorescence in situ hybridization. Protection of A. thaliana seedlings inoculated with NRRL B-24137 against B. cinerea was then evaluated. Parts of the mechanisms involved in the systemic protection against B. cinerea were evaluated using known mutants of genes involved in jasmonate (JA)/ethylene (ET)/salicylic acid (SA) signaling. Other Arabidopsis mutants, AtrhbohD-3, AtrhbohF-3, and ups1-1 were also screened to determine other parts of the mechanisms involved. Results The results showed that the strain NRRL B-24137 colonized, epi- and endophytically, the roots of Arabidopsis seedlings but the strain was not a systemic colonizer during the time of the experiment. The strain NRRL B-24137 also reduced B. cinerea symptoms and the protection was linked to known mechanisms of induced systemic resistance (ISR; JA/ET signaling), as well as to functionality of AtrbohF oxidase and of UPS1. Crosstalk between ET/JA and SA signaling could also be involved. Conclusions The isolate NRRL B-24137, after colonizing the root systems of A. thaliana, induces an ISR against B. cinerea, which is JA/ET dependent, but could also require SA crosstalk and protection could also require NAPDH oxidases and UPS1 functionalities.

Background and aim There is currently a gap of knowledge regarding whether some beneficial bacteria isolated from desert soils can colonize epi- and endophytically plants of temperate regions. In this study, the early steps of the colonization process of one of these bacteria, Saccharothrix algeriensis NRRL B-24137, was studied on grapevine roots to determine if this beneficial strain can colonize a non-natural host plant. An improved method of fluorescence in situ hybridization (FISH), the double labeling of oligonucleotide probes (DOPE)-FISH technique was used to visualize the colonization behavior of such bacteria as well as to determine if the method could be used to track microbes on and inside plants. Methods A probe specific to Saccharothrix spp. was firstly designed. Visualization of the colonization behavior of S. algeriensis NRRL B-24137 on and inside roots of grapevine plants was then carried out with DOPE-FISH microscopy. Results The results showed that 10 days after inoculation, the strain could colonize the root hair zone, root elongation zone, as well as root emergence sites by establishing different forms of bacterial structures as revealed by the DOPE-FISH technique. Further observations showed that the strain could be also endophytic inside the endorhiza of grapevine plants. Conclusions Taking into account the natural niches of this beneficial strain, this study exemplifies that, in spite of its isolation from desert soil, the strain can establish populations as well as subpopulations on and inside grapevine plants and that the DOPE-FISH tool can allow to detect it.

The production of the 16-membered macrolide antibiotic, spiramycin, in Streptomyces ambofaciens is inhibited by glucose, 2-deoxyglucose and inorganic phosphate. The role of intracellular ATP content and phosphorylated metabolites as common regulating signals of both glucose and phosphate inhibitory effects is discussed. Two enzymatic targets of the effect of phosphate on spiramycin biosynthesis were studied. Valine dehydrogenase, the first enzyme of valine catabolism (supplier of aglycone spiramycin precursors), and alkaline phosphatase, which cleaves phosphorylated intermediates, were repressed in the presence of excess phosphate.