A new dithiolopyrrolone antibiotic triggered by a long fermentation of Saccharothrix algeriensis NRRL B-24137 in sorbic acid-amended medium.

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.

Effective biosynthesis of benzoyl-pyrrothine dithiolopyrrolone antibiotic by cinnamic acid-precursor addition in culture of Saccharothrix algeriensis NRRL B-24137.

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.

Development of a novel quantitative PCR assay as a measurement for the presence of geosmin-producing fungi.

AIMS: To provide an efficient technique for monitoring the off-flavoured fungal compound geosmin. METHODS AND RESULTS: 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'. CONCLUSIONS: Penicillium spp., genomic DNA level can provide an efficient way to quantify geosmin. SIGNIFICANCE AND IMPACT OF THE STUDY: This particular qPCR technique can be utilized in numerous food industries, for the timely detection and monitoring of geosmin contamination.

A novel hydroxamic acid-containing antibiotic produced by a Saharan soil-living Streptomyces strain.

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.

Characterization and antagonistic properties of Streptomyces strains isolated from Saharan soils, and evaluation of their ability to control seedling blight of barley caused by Fusarium culmorum.

UNLABELLED: During a screening for potential plant disease control actinomycetes, a total of 133 strains were isolated from Saharan soil samples of seven Algerian regions by dilution technique on chitin-vitamins agar medium. Screening for antagonistic properties using streak assay method showed that 25% of isolates demonstrated strong activities against a wide range of plant pathogenic fungi. Due to their strong anti-Fusarium activities, six of these isolates were selected and subsequently related to Streptomyces species by polyphasic analysis. These isolates were evaluated for their biocontrol ability against Fusarium culmorum, a serious pathogenic fungus of cereals crops related to damping-off and seedling blight resulting in yield loss. Barley seeds were chosen as cereal plant model. Surface bacterized seeds with TW3, RI3 and TW2 strains expressed the highest performances and permit to reduce significantly both the disease occurrence on seedlings (62-76%) and the extent of seedling blight symptoms (over than 95%). However, a negative effect on plant establishment was observed for RI3 treatment. SIGNIFICANCE AND IMPACT OF THE STUDY: The genus Fusarium is considered to be one of the most problematic phytopathogenic fungi for crop culture worldwide. Inside this genus, F. culmorum is the aetiological agent of seedling blight in various monocotyledonous plants such as barley and cause extensive yield and quality losses in humid and semi-humid regions. Biological control may be a successful alternative to chemical control, particularly with the controversy surrounding the use of the fungicides and the limited obtained results to control F. culmorum. This study highlights the effectiveness of some antagonistic Streptomyces isolated from Algerian Saharan soils to control F. culmorum by the reduction in disease occurrence and disease severity suggesting their use on microbial biocontrol formulation against soilborne diseases.

In vitro and in vivo antagonism of actinomycetes isolated from Moroccan rhizospherical soils against Sclerotium rolfsii: a causal agent of root rot on sugar beet (Beta vulgaris L.).

AIMS: To evaluate the ability of the isolated actinomycetes to inhibit in vitro plant pathogenic fungi and the efficacy of promising antagonistic isolates to reduce in vivo the incidence of root rot induced by Sclerotium rolfsii on sugar beet. METHODS AND RESULTS: Actinomycetes isolated from rhizosphere soil of sugar beet were screened for antagonistic activity against a number of plant pathogens, including S. rolfsii. Ten actinomycetes out of 195 screened in vitro were strongly inhibitory to S. rolfsii. These isolates were subsequently tested for their ability to inhibit sclerotial germination and hyphal growth of S. roflsii. The most important inhibitions were obtained by the culture filtrate from the isolates J-2 and B-11, including 100% inhibition of sclerotial germination and 80% inhibition of hyphal growth. These two isolates (J-2 and B-11) were then screened for their ability to protect sugar beet against infection of S. rolfsii induced root rot in a pot trial. The treatment of S. rolfsii infested soil with a biomass and culture filtrate mixture of the selected antagonists reduced significantly (P < or = 0.05) the incidence of root rot on sugar beet. Isolate J-2 was most effective and allowed a high fresh weight of sugar beet roots to be obtained. Both antagonists J-2 and B-11 were classified as belonging to the genus Streptomyces species through morphological and chemical characteristics as well as 16S rDNA analysis. CONCLUSION: Streptomyces isolates J-2 and B-11 showed a potential for controlling root rot on sugar beet and could be useful in integrated control against diverse soil borne plant pathogens. SIGNIFICANCE AND IMPACT OF THE STUDY: This investigation showed the role, which actinomycete bacteria can play to control root rot caused by S. rolfsii, in the objective to reduce treatments with chemical fungicides.

Expression of pyrrothine N-acyltransferase activities in Saccharothrix algeriensis NRRL B-24137: new insights into dithiolopyrrolone antibiotic biosynthetic pathway.

AIMS: The hypothetical dithiolopyrrolone biosynthetic pathway includes a final step of pyrrothine nucleus acylation. The presence of an enzymatic activity catalysing this reaction was investigated in Saccharothrix algeriensis NRRL B-24137. To understand the effect exerted by organic acids on the level of dithiolopyrrolone production, their influence on enzymatic expression was studied. METHODS AND RESULTS: The transfer of acetyl-CoA or benzoyl-CoA on pyrrothine was assayed in the cell-free extract of Sa. algeriensis NRRL B-24137. This study reports the presence of an enzymatic activity catalysing this reaction that was identified as either pyrrothine N-acetyltransferase or N-benzoyltransferase. The stimulation of benzoyl-pyrrothine (BEP) production by addition of benzoic acid at 1.25 mmol l(-1) into the culture medium was demonstrated, and results showed that under the same conditions of growth, pyrrothine N-benzoyltransferase specific activity was doubled. CONCLUSIONS: This study shows that BEP production is enhanced in the presence of benzoic acid partly because of an induction of pyrrothine N-benzoyltransferase. SIGNIFICANCE AND IMPACT OF THE STUDY: The antitumor and antibiotic properties of dithiolopyrrolones are related to their variable acyl groups. New insights into regulation of biosynthetic pathway, especially the step of pyrrothine acylation, could lead after further studies to yield improvement and to selective production of dithiolopyrrolones with new biological activities.

Mycoflora and ochratoxin A producing strains of Aspergillus in Algerian wheat.

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).

Isolation of Bacillus spp. from Thai fermented soybean (Thua-nao): screening for aflatoxin B1 and ochratoxin A detoxification.

AIMS: To study the interaction between Bacillus spp. and contaminating Aspergillus flavus isolated strains from Thai fermented soybean in order to limit aflatoxin production. To study the detoxification of aflatoxin B(1) (AFB(1)) and ochratoxin A (OTA) by Bacillus spp. in order to find an efficient strain to remove these toxins. METHODS AND RESULTS: One A. flavus aflatoxin-producing strain and 23 isolates of Bacillus spp. were isolated from soybean and fresh Thua-nao collected from the north of Thailand. Inhibition studies of A. flavus and A. westerdijkiae NRRL 3174 (reference strain) growth by all isolates of Bacillus spp. were conducted by dual culture technique on agar plates. These isolates were also tested for AFB(1) and OTA detoxification ability on both solid and liquid media. Most of the strains were able to detoxify aflatoxin but only some of them could detoxify OTA. CONCLUSIONS: One Bacillus strain was able to inhibit growth of both Aspergillus strains and to remove both mycotoxins (decrease of 74% of AFB(1) and 92.5% of OTA). It was identified by ITS sequencing as Bacillus licheniformis. The OTA decrease was due to degradation in OTalpha. Another Bacillus strain inhibiting both Aspergillus growth and detoxifying 85% of AFB(1) was identified as B. subtilis. AFB(1) decrease has not been correlated to appearance of a degradation product. SIGNIFICANCE AND IMPACT OF THE STUDY: The possibility to reduce AFB(1) level by a strain from the natural flora is of great interest for the control of the quality of fermented soybean. Moreover, the same strain could be a source of efficient enzyme for OTA degradation in other food or feeds.

Occurrence of pathogenic fungal species in Tunisian vineyards.

Mycotoxins are secondary metabolites produced by filamentous fungi detected in food, such are grapes. OTA was evaluated in ten handle musts from different Tunisian vineyard. This mycotoxin was found at levels 1.1 mug/L to 4.3 mug/L. A survey was conducted to assess the contamination of the Tunisian vineyard with pathogenic fungal species, in particular those responsible of the OTA production. The results were evaluated for the first time in parcels cultivated in the North, in the Centre and in the South of the country. Italia Muscate and Superior Seedless varieties were concerned at three developmental stages of the berry, setting, veraison and maturity. Carigon variety was used as positive control for musts contaminating by OTA. The main fungal species isolated were Aspergillus spp. (33.32%), Botrytis cinerea (23.32%), Alternaria spp. (12.80%), Cladosporium spp. (10.59%) and Penicillium spp. (8.3%). The isolates of the Aspergillus genus were identified as Aspergillus niger aggregate (77%), Aspergillus carbonarius (15%) and Aspergillus flavus (8%). Their presence was characterized by a significant decrease in the Centre during the veraison and a slight increase in the North and the South during the maturity stage. Furthermore, when comparing Superior Seedless and Italia Muscate cultivated in the same area, the aspergilli were particularly less abundant at the setting stage in the case of Superior Seedless. There is no correlation between the OTA amount in musts and the contamination by Aspergillus species in different vineyards and for grape varieties studied.

Black aspergilli and ochratoxin A production in French vineyards.

A survey on the occurrence on grape of black Aspergillus species and their capability to produce ochratoxin A (OTA) was conducted in France over three years (2001-2003) in 10 vineyards from four winemaking regions with different geographical locations and climatic conditions. During 2001 and 2002, from setting to harvest, the total numbers of fungal isolates were respectively 721 and 711 increasing in 2003 to reach 1035. The Aspergillus genus was essentially represented by Section Nigri (99%) and it was predominant (80%+/-4.6) when compared to Penicillium (20%+/-4.6). Regardless of sampling year, 32.5% (+/-sigma=1.26) of the fungal isolates were OTA producers and 93% (+/-sigma=2.65) belonging to black aspergilli. The ochratoxigenic potential of the isolates and their occurrence on grapes revealed that Aspergillus carbonarius was the main OTA producer (up to 37.5 mug/g). At harvest time, the fungal population was maximal and this was the most critical period influencing OTA contamination. Grapes from Languedoc-Roussillon region were most infested with ochratoxigenic fungi and had the highest concentrations of OTA (up to 2.8 ng/g).

Conidia of black aspergilli as new biological adsorbents for ochratoxin A in grape juices and musts.

Biological removal of ochratoxin A (OTA) by living and heat-treated dead conidia of black Aspergillus isolates representing the species Aspergillus niger, Aspergillus carbonarius, and Aspergillus japonicus in synthetic and natural grape juices was found to be a two-stage phenomenon. Several lines of evidence suggest that the first observed stage was passive, metabolism was not required, and OTA adsorption on conidia of black aspergilli could be involved. This removal was fast, without delay just after conidial inoculation both in synthetic and natural grape juices. Moreover, even nonviable, heat-treated conidia were capable of removing OTA. Finally, no OTA degradation products were detected. In the second observed stage, removal of OTA was linked to degradation by live conidia only. Ochratoxin alpha, a degradation product of OTA, was detected in the medium after incubation for 30 and 14 h for biseriate (A. niger and A. carbonarius) and uniseriate (A. japonicus) black aspergilli, respectively, when well-developed mycelium appeared. Comparisons between the three black Aspergillus isolates tested showed that A. carbonarius detoxified grape juice most effectively. However, this species often produces OTA. A. niger and A. japonicus isolates were also effective and because those species are not systematically OTA producers, they could be interesting for further OTA detoxification processes in grape juices and musts.

Antibiotic resistance gene cassettes derived from the omega interposon for use in E. coli and Streptomyces.

Three antibiotic resistance gene cassettes, derived from the omega interposon (Prentki and Krisch (1984) Gene 29, 303-313) were constructed. These cassettes carry different antibiotic resistance genes, conferring resistance to geneticin, hygromycin or viomycin, flanked by short inverted repeats containing transcription and translation termination signals and synthetic polylinkers. These cassettes were designated omega aac, omega hyg and omega vph. Resistance phenotypes conferred by these constructions are selectable in E. coli and Streptomyces. These cassettes can be used for insertional mutagenesis or for vector construction.

Effect of the bacteriocin carnocin CP5 and of the producing strain Carnobacterium piscicola CP5 on the viability of Listeria monocytogenes ATCC 15313 in salt solution, broth and skimmed milk, at various incubation temperatures.

Carnobacterium piscicola CP5, isolated from French mould-ripened soft-cheese, produced a bacteriocin named carnocin CP5 in a wide range of incubation temperatures, from 4 degrees C to 30 degrees C. The ability of a crude bacteriocin, of a partially-purified form, and of the producer strain to inhibit growth of Listeria monocytogenes ATCC 15313 was examined in salt solution, broth and skimmed milk between 4 degrees C and 30 degrees C. When carnocin CP5 was added to a L. monocytogenes ATCC 15313 culture, an adsorption on cells and a bactericidal effect with a cell lysis occurred. At 30 degrees C, with carnocin CP5 or with C. piscicola CP5, a transitory bactericidal effect was observed. Subsequent experiments at 4 degrees C, 7 degrees C or 15 degrees C, showed a more prolonged bactericidal effect. Thus at 7 degrees C, partially-purified carnocin CP5 reduced an initial population level of L. monocytogenes of 10(3) cfu/ml to non-detectable level within 7 days. However, in some cases, with extended incubation, the carnocin CP5 effect was no longer visible, the L. monocytogenes population grew again. This phenomenon was probably due to the presence of a sub-population of bacteriocin-resistant variants.

Mode of action, purification and amino acid sequence of plantaricin C19, an anti-Listeria bacteriocin produced by Lactobacillus plantarum C19.

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.

Identification of two putative acyltransferase genes potentially implicated in dithiolopyrrolone biosyntheses in Saccharothrix algeriensis NRRL B-24137.

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.

Taxonomy and chemical characterization of new antibiotics produced by Saccharothrix SA198 isolated from a Saharan soil.

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 (1)H and (14)C 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.

[Antimicrobial activity of a Saharan Streptomyces spp. PAL111 strain against various clinical and toxinogenic microorganisms resistant to antibiotics]. / Activité antimicrobienne de Streptomyces sp. PAL111 d'origine saharienne contre divers microorganismes cliniques et toxinogènes résistants aux antibiotiques.

OBJECTIVE: Study of the taxonomy and the biological activity of the actinomycete strain PAL111 against several pathogenic and toxigenic microorganisms for humans, and resistant to many antibiotics. MATERIALS AND METHODS: The taxonomic study of isolate PAL111 is carried out on the basis of phenotypic and molecular characteristics. The tests against the pathogenic microorganisms are realized on ISP-2 and Bennett media. The kinetics of antibiotic production was investigated on ISP-2 medium. The antibiotic is highlighted by bioautography and chemical revelations, and then purified by chromatography on thick layer of silica gel and Sephadex LH20 column. The minimum inhibitory concentrations (MIC) were determined against pathogenic microorganisms. RESULTS: The phenotypic and molecular studies showed that the isolate PAL111 is closely related to the type strain of Streptomyces ambofaciens. It showed a strong activity against Candida albicans, filamentous fungi, and Gram-positive and Gram-negative bacteria. The optimal antibiotic production was observed at the end of the exponential phase of growth and at the beginning of the decline phase. The bioautography tests showed the presence of an antibiotic with both antibacterial and antifungal activities. This antibiotic is a hydrophilic amino-glycoside compound. The MIC were observed between 2 and 20µg/mL for yeasts, 10 and 50µg/mL for filamentous fungi, 2 and 10µg/mL for Gram-positive bacteria, and 20 and 75µg/mL for Gram-negative bacteria. CONCLUSION: The strong activity of isolate PAL111 against the pathogenic microorganisms and the polar characteristic of the produced antibiotic could encourage further studies on this bioactive molecule.

Chemical composition and in vitro antimicrobial and antioxidant activities of Citrus aurantium l. flowers essential oil (Neroli oil).

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.

Review of mycotoxin reduction in food and feed: from prevention in the field to detoxification by adsorption or transformation.

Mycotoxins are secondary metabolites present worldwide in agricultural commodities and produced by filamentous fungi that cause a toxic response (mycotoxicosis) when ingested by animals. Prevention of mycotoxicoses includes pre- and post-harvest strategies. The best way to reduce the mycotoxin content in food and feed is the prevention of mycotoxin formation in the field, but this is often not sufficient, so other methods are needed. To decontaminate and/or detoxify mycotoxin-contaminated food and feed, the most prevalent approach in the feed industry is the inclusion of sorbent materials in the feed thus obtaining more or less selective removal of toxins by adsorption during passage through the gastrointestinal tract. Another reliable approach is to add enzymes or microorganisms capable of detoxifying some mycotoxins. Through a comprehensive review of published reports on the strategies for mycotoxin removal, this present work aims to update our understanding of mycotoxin removal. It provides an insight into the detoxification of mycotoxin present in food and feed. In the future, more emphasis needs to be placed on adsorption of mycotoxins in the gastrointestinal tract. Concerning the enzymatic transformation of mycotoxins, further efforts are required in understanding detoxification reactions, the toxicity of transformation products and in the characterization of enzymes responsible for transformations.