Correction: Ibrahimi et al. Marine Actinobacteria: Screening for Predation Leads to the Discovery of Potential New Drugs against Multidrug-Resistant Bacteria. Antibiotics 2020, 9, 91.

In the original publication, there was a mistake in Figure 1b as published [...].

Microbial intervention improves pollutant removal and semi-liquid organo-mineral fertilizer production from olive mill wastewater sludge and rock phosphate.

Olive mill wastewater sludge (OMWS) represents a residual pollutant generated by the olive oil industry, often stored in exposed evaporation ponds, leading to contamination of nearby land and water resources. Despite its promising composition, the valorization of OMWS remains underexplored compared to olive mill wastewater (OMW). This study aims to identify potent native microbial species within OMWS suitable for bioremediation and its transformation into a high-value organic fertilizer. The microbial screening, based on assessing OMWS tolerance and phosphate solubilization properties in vitro, followed by a singular inoculation using a mixture of OMWS and rock phosphate (RP). Identification of FUN 06 (Galactomyces Geotrichum), a fungal species, employed as an inoculant in the treatment of sterile OMWS supplemented with RP. Results demonstrate that fungal inoculation notably diminished OMWS phytotoxicity while enhancing its physicochemical parameters, nutrient concentrations, and removal of toxic organic compounds by up to 90% compared to the control, and enhances plant growth, offering a sustainable approach to tackle environmental concerns. Additionally, metataxonomic analysis unveiled FUN 06's propensity to enhance the presence of microbial species engaged in pollutant degradation. However, higher RP dosage (10%) appeared to adversely affect bioprocess efficiency, suggesting a potential dose-related effect. Overall, FUN 06, isolated from OMWS evaporation ponds, shows promise for effective bioremediation and sustainable reuse. In fact, our results indicate that targeted microbial inoculation stands as an effective strategy for mitigating pollutants in OMWS, facilitating its conversion into a nutrient-rich organo-mineral fertilizer suitable for direct use, promoting its beneficial reuse in agriculture, thereby presenting a promising avenue for olive oil waste management.

Mycorrhizas and Trichoderma fungi increase the accumulation of secondary metabolites in grain legume leaves and suppress foliar diseases in field-grown conditions of the humid forest of Cameroon.

BACKGROUND: Arbuscular mycorrhizal and Trichoderma fungi alter the synthesis of secondary metabolites of plants and confer tolerance from pathogens attacks. However, there is less supportive evidence from on-field studies confirming the above-mentioned hypothesis, particularly for the humid forest zone of Cameroon where pathogens are important sources of yield losses for legumes such as soybean and common bean. MATERIALS AND METHODS: We evaluated the impacts of mycorrhiza isolates of Rhizophagus intraradices (Ri) and Trichoderma asperellum (Ta) fungi and their co-inoculations (Ta x Ri) in the synthetizing of leaves secondary metabolites, foliar disease symptoms, growth, N and P uptake, and yields of three genotypes of soybean (TGx 1485-1D, TGx 1990-93 F, and TGx 1990-97 F) and common beans (NUA-99, DOR-701, and PNN) under field conditions of Cameroon. RESULTS: We found that common bean plants showed a lower foliar infection rate but a higher increase in root colonization intensity, shoot dry weight, and N and P uptakes than soybeans when inoculated with Ri and Ta treatment. However, the grain yield of soybean soybean was higher (2000 kg ha 1) than the common bean plants for the Ri × Ta treatment. The soybean genotype TGx 1990-93F had increased root colonization intensity and the lowest foliar infection rate, making it stronger and tolerant to pathogen attacks when co-inoculated with Ri × Ta fungi (F). Bean plants inoculated with Ri and the co-inoculated with Ri × Ta demonstrated lower symptoms of foliar attack, and increased root colonization, particularly the PNN variety. The total amino acid and proline accumulations were higher for soybean than common bean plants due to fungi inoculations, and soybean genotypes accumulated more excellent contents of amino acid and proline in the control (10.1 mg g- 1 fwt) that significantly increased under the Ri × Ta inoculation (13.4 mg g- 1 fwt). CONCLUSIONS: Common bean plants inoculated with Ta and Ri fungi accumulated higher phenolic compounds in their leaves that aided them in overcoming the pathogen attacks than soybean plants.

Improving the efficiency of phosphate rocks combined with phosphate solubilizing Actinomycetota to increase wheat growth under alkaline and acidic soils.

Low availability of phosphorus (P) in both acidic and alkaline soils is a major problem for sustainable improvement in wheat crops yield. Optimization of crops productivity can be achieved by increasing the bioavailability of P by phosphate solubilizing Actinomycetota (PSA). However, their effectiveness may vary with changing agro-climatic conditions. In this regard, a greenhouse experiment was conducted to assess the interaction inoculation of five potential PSA (P16-P18-BC3-BC10 and BC11) and RPs (RP1- RP2-RP3 and RP4) on the growth and yield of wheat crop in unsterilized P- deficient alkaline and acidic soils. Their performance was compared with single super phosphate (TSP) and reactive RP (BG4). The in-vitro tests showed that all PSA colonize wheat root and form a strong biofilm except Streptomyces anulatus strain P16. Our findings revealed that all PSA significantly improve the shoot/root dry weights, spike biomass, chlorophyll contents as well as nutrients uptake in plants fertilized with RP3 and RP4. However, the combined application of Nocardiopsis alba BC11 along with RP4 in alkaline soil, was effective in optimizing wheat yield attributes and improve the yield biomass up to 19.7% as compared to the triple superphosphate (TSP). This study supports the view that the inoculation with Nocardiopsis alba BC11 has a broad RP solubilization and could alleviate the agricultural losses due to P limitation in acidic and alkaline soils.

Olive mill waste sludge: From permanent pollution to a highly beneficial organic biofertilizer: A critical review and future perspectives.

Olive mill wastewater sludge (OMWS) is a by-product of the olive extraction process that is attracting substantial attention due to its extremely hazardous effects on aquatic and terrestrial ecosystems. OMWS is a product of the common disposal method of olive oil mill wastewater (OMWW) that accumulates in evaporation ponds. It is estimated that approximately 10 × 106 m3 of OMWS is generated worldwide each year. OMWS is characterized by its significantly variable physicochemical properties and organic pollutant constituents, such as phenols and lipids, which are dependent upon the environmental features of the receiving ponds. Nonetheless, many related studies have recognized the biofertilizer potential of this sludge owing to its high mineral nutrient and organic matter load. OMWS exhibits promising valorization potential in several fields, including agriculture and energy production. Compared to those of OMWW, studies of OMWS are still lacking concerning its composition and characteristics, which are necessary for the future implementation of efficient valorization strategies. The main purpose of this review paper is to fill the gap that exists in the literature by providing a critical analysis of the available data on OMWS production, distribution, characteristics, and properties. Additionally, this work sheds light on important factors affecting OMWS properties, including the variability of the indigenous microbial communities regarding bioremediation. Finally, this review addresses the current and future valorization routes, from detoxification to the development of promising applications in agriculture, energy, and the environment, which could have significant socioeconomic implications for low-income Mediterranean countries.

Grain Legume Yield Responses to Rhizobia Inoculants and Phosphorus Supplementation Under Ghana Soils: A Meta-Synthesis.

A discrete number of studies have been conducted on the effects of rhizobia (Rhz) inoculants, phosphorus (P) management, and combined application of Rhz and P fertilizer on the enhancement of grain legume yield across soils of Ghana and elsewhere. However, the extent to which the various inoculated Rhz strains, P application, and combined application of Rhz + P studies contribute to improving yield, performed on a comprehensive analysis approach, and profit farmers are yet to be understood. This study reviewed different experimental studies conducted on soybean (Glycine max (L.) Merr.), cowpea (Vigna unguiculata [L.] Walp), and groundnut (Arachis hypogaea [L.]) to which Rhz inoculants, P supplements, or Rhz + P combination were applied to improve the yield in Ghana. Multiple-step search combinations of published articles and multivariate analysis computing approaches were used to assess the effects of Rhz inoculation, P application, or both application of Rhz and P on yield variation. The random forest (RF) regression model was further employed to quantify the relative importance of various predictor variables on yield. The meta-analysis results showed that cowpea exhibited the highest (61.7%) and groundnut (19.8%) the lowest average yield change. The RF regression model revealed that the combined application of Rhz and P fertilizer (10.5%) and Rhz inoculation alone (7.8%) were the highest explanatory variables to predict yield variation in soybean. The Rhz + P combination, Rhz inoculation, and genotype wang-Kae explained 11.6, 10.02, and 8.04% of yield variability for cowpea, respectively. The yield in the inoculated plants increased by 1.48-, 1.26-, and 1.16-fold when compared to that in the non-inoculated cowpea plants following inoculation with BR 3299, KNUST 1002, and KNUST 1006 strains, respectively. KNUST 1006 strain exhibited the highest yield increase ratio (1.3-fold) in groundnut plants. Inoculants formulation with a viable concentration of 109 cells g-1 and a minimum inoculum rate of 1.0 × 106 cells seed-1 achieved the highest average yield change for soybean but not for cowpea and groundnut. The meta-analysis calls for prospective studies to investigate the minimum rate of bacterial cells required for optimum inoculation responses in cowpea and groundnut.

Multifunctional role of Actinobacteria in agricultural production sustainability: A review.

The growing interest in low-input agriculture in recent years has focused the use of microbial biofertilizers to improve plant growth and yield through a better mobilization of indigenous source of key nutrients such as nitrogen, phosphorus, potassium etc. In this context, soil microorganisms especially Actinobacteria might play an important role. With their multifunctional activities, they are involved in nutrient cycling, soil quality and crop productivity as well as plant health which make them not only the eco-friendly alternative for agriculture but also for humankind. Bearing this in mind, it is primordial to further explore the special link between these microorganisms and soil -plant ecosystems. Therefore, this review discusses the importance of Actinobacteria as microbial biofertilizers and highlights the future needs and challenges for using them for sustaining crop. The patents and scientific literature analysis from 2000 to 2020 show that 16 patents claiming Actinobacteria as biocontrol or biofertilizer in agriculture and 949 indexed research articles related to Actinobacteria effect on plant growth and phosphate solubilization have been published. Furthermore, Actinobacteria ability to increase growth and yield of staple crops such as wheat maize, tomato, rice, and chickpea plant have been highlighted. Much more effort and progress are expected in the industrial development of actinobacterial bioinoculants as areas such as synthetic biology and nano-biotechnology advance.

The potential of facultative predatory Actinomycetota spp. and prospects in agricultural sustainability.

Actinomycetota in the phylum of bacteria has been explored extensively as a source of antibiotics and secondary metabolites. In addition to acting as plant growth-promoting agents, they also possess the potential to control various plant pathogens; however, there are limited studies that report the facultative predatory ability of Actinomycetota spp. Furthermore, the mechanisms that underline predation are poorly understood. We assessed the diversity of strategies employed by predatory bacteria to attack and subsequently induce the cell lysing of their prey. We revisited the diversity and abundance of secondary metabolite molecules linked to the different predation strategies by bacteria species. We analyzed the pros and cons of the distinctive predation mechanisms and explored their potential for the development of new biocontrol agents. The facultative predatory behaviors diverge from group attack "wolfpack," cell-to-cell proximity "epibiotic," periplasmic penetration, and endobiotic invasion to degrade host-cellular content. The epibiotic represents the dominant facultative mode of predation, irrespective of the habitat origins. The wolfpack is the second-used approach among the Actinomycetota harboring predatory traits. The secondary molecules as chemical weapons engaged in the respective attacks were reviewed. We finally explored the use of predatory Actinomycetota as a new cost-effective and sustainable biocontrol agent against plant pathogens.

Potential Role and Utilization of Plant Growth Promoting Microbes in Plant Tissue Culture.

Plant growth promoting microbes (PGPMs) play major roles in diverse ecosystems, including atmospheric nitrogen fixation, water uptake, solubilization, and transport of minerals from the soil to the plant. Different PGPMs are proposed as biofertilizers, biostimulants, and/or biocontrol agents to improve plant growth and productivity and thereby to contribute to agricultural sustainability and food security. However, little information exists regarding the use of PGPMs in micropropagation such as the in vitro plant tissue culture. This review presents an overview of the importance of PGPMs and their potential application in plant micropropagation. Our analysis, based on published articles, reveals that the process of in vitro classical tissue culture techniques, under strictly aseptic conditions, deserves to be reviewed to allow vitroplants to benefit from the positive effect of PGPMs. Furthermore, exploiting the potential benefits of PGPMs will lead to lessen the cost production of vitroplants during micropropagation process and will make the technique of plant tissue culture more efficient. The last part of the review will indicate where research is needed in the future.

The Screening of Potassium- and Phosphate-Solubilizing Actinobacteria and the Assessment of Their Ability to Promote Wheat Growth Parameters.

Soil fertility and plant nutrition require an adequate management of essential macronutrients such as potassium (K) and phosphorus (P), which are mandatory for plant development. Bioleaching of K and P bearing minerals improves their chemical weathering and increases the performance of the biofertilization strategies. In this study, in vitro and greenhouse experiments were carried out to investigate P and K solubilization traits of nine Actinobacteria (P13, P14, P15, P16, P17, P18, BC3, BC10, and BC11) under fertilization with rock phosphate (RP). K and P solubilization were evaluated on Alexandrov and NBRIP media containing mica and six RP samples, respectively. The actinobacterial strains were able to solubilize K in Alexandrov medium supplemented with RP. However, when soluble P was used instead of RP, only four strains of Actinobacteria (Streptomyces alboviridis P18-Streptomyces griseorubens BC3-Streptomyces griseorubens BC10 and Nocardiopsis alba BC11) solubilized K. The solubilization values of K ranged from 2.6 to 41.45 mg/L while those of P varied from 0.1 to 32 mg/L. Moreover, all strains were able to produce IAA, siderophore, HCN, and ammonia and significantly improved the germination rate and the vigor index of wheat. The pot experiments revealed that four strains (Streptomyces alboviridis P18, Streptomyces griseorubens BC3, Streptomyces griseorubens BC10, and Nocardiopsis alba BC11) significantly improved the growth parameters of wheat, namely root length (1.75-23.84%), root volume (41.57-71.46%), root dry weight (46.89-162.41%), shoot length (8.92-23.56%), and shoot dry weight (2.56-65.68%) compared to the uninoculated control. These findings showed that Streptomyces griseorubens BC10 and Nocardiopsis alba BC11 are promising candidates for the implementation of efficient biofertilization strategies to improve soil fertility and plant yield under rock P and rock K fertilization.

Assessment of actinobacteria use in the elimination of multidrug-resistant bacteria of Ibn Tofail hospital wastewater (Marrakesh, Morocco): a chemometric data analysis approach.

The efficiency of the treatment of hospital wastewater by actinobacteria was investigated using two chemometric data analysis methods. Six strains of multi-resistant bacteria isolated from Marrakesh hospital wastewater and four strains of antagonistic actinobacteria isolated from Moroccan marine environment were characterized by fatty acids released as methyl esters by thermochemolysis-GC/MS. The hierarchical cluster analysis (HCA) and the principal component analysis (PCA) were used to correlate fatty acids (FA) distributions within strains. HCA allowed to discriminate between bacteria and actinobacteria. A lower Euclidean distance is noted for bacteria. With PCA, linear and branched-chained FAs correlated with bacteria whereas mono unsaturated FAs correlated more specifically with Gram (-) bacteria. Terminally branched-chained FAs correlated most likely with actinobacteria. A co-culture of actinobacteria and bacteria monitored during 15 days demonstrated the efficiency of the biological treatment for 2 of the 4 studied actinobacteria. The effect is more important on Gram-negative bacteria. Antagonistic actinobacteria seem to be poorly efficient against Gram-positive bacteria.

A Molecule of the Viridomycin Family Originating from a Streptomyces griseus-Related Strain Has the Ability to Solubilize Rock Phosphate and to Inhibit Microbial Growth.

Some soil-borne microorganisms are known to have the ability to solubilize insoluble rock phosphate and this process often involves the excretion of organic acids. In this issue, we describe the characterization of a novel solubilizing mechanism used by a Streptomyces strain related to Streptomyces griseus isolated from Moroccan phosphate mines. This process involves the excretion of a compound belonging to the viridomycin family that was shown to play a major role in the rock phosphate bio weathering process. We propose that the chelation of the positively charged counter ions of phosphate constitutive of rock phosphate by this molecule leads to the destabilization of the structure of rock phosphate. This would result in the solubilization of the negatively charged phosphates, making them available for plant nutrition. Furthermore, this compound was shown to inhibit growth of fungi and Gram positive bacteria, and this antibiotic activity might be due to its strong ability to chelate iron, a metallic ion indispensable for microbial growth. Considering its interesting properties, this metabolite or strains producing it could contribute to the development of sustainable agriculture acting as a novel type of slow release bio-phosphate fertilizer that has also the interesting ability to limit the growth of some common plant pathogens.

Microbial Community Succession and Organic Pollutants Removal During Olive Mill Waste Sludge and Green Waste Co-composting.

Olive mill wastewater sludge (OMWS) is the main by-product of the olive industry. OMWS is usually dumped in landfills without prior treatment and may cause several eco-environmental hazards due to its high toxicity, which is mainly attributed to polyphenols and lipids. OMWS is rich in valuable biocompounds, which makes it highly desirable for valorization by composting. However, there is a need to understand how microbial communities evolve during OMWS composting with respect to physicochemical changes and the dynamics of pollutant degradation. In this study, we addressed the relationship between microbial community, physicochemical variations and pollutants degradation during the co-composting of OMWS and green wastes using metagenomic- and culture-dependent approaches. The results showed that in raw OMWS, Pichia was the most represented genus with almost 53% of the total identified fungal population. Moreover, the bacteria that dominated were Zymobacter palmae (20%) and Pseudomonas sp. (19%). The addition of green waste to OMWS improved the actinobacterial diversity of the mixture and enhanced the degradation of lipids (81.3%) and polyphenols (84.54%). Correlation analysis revealed that Actinobacteria and fungi (Candida sp., Galactomyces sp., and Pichia manshurica) were the microorganisms that had the greatest influence on the composting process. Overall, these findings provide for the first time some novel insights into the microbial dynamics during OMWS composting and may contribute to the development of tailored inoculum for process optimization.

Exploiting Biological Nitrogen Fixation: A Route Towards a Sustainable Agriculture.

For all living organisms, nitrogen is an essential element, while being the most limiting in ecosystems and for crop production. Despite the significant contribution of synthetic fertilizers, nitrogen requirements for food production increase from year to year, while the overuse of agrochemicals compromise soil health and agricultural sustainability. One alternative to overcome this problem is biological nitrogen fixation (BNF). Indeed, more than 60% of the fixed N on Earth results from BNF. Therefore, optimizing BNF in agriculture is more and more urgent to help meet the demand of the food production needs for the growing world population. This optimization will require a good knowledge of the diversity of nitrogen-fixing microorganisms, the mechanisms of fixation, and the selection and formulation of efficient N-fixing microorganisms as biofertilizers. Good understanding of BNF process may allow the transfer of this ability to other non-fixing microorganisms or to non-leguminous plants with high added value. This minireview covers a brief history on BNF, cycle and mechanisms of nitrogen fixation, biofertilizers market value, and use of biofertilizers in agriculture. The minireview focuses particularly on some of the most effective microbial products marketed to date, their efficiency, and success-limiting in agriculture. It also highlights opportunities and difficulties of transferring nitrogen fixation capacity in cereals.

Marine Actinobacteria: Screening for Predation Leads to the Discovery of Potential New Drugs against Multidrug-Resistant Bacteria.

Predatory bacteria constitute a heterogeneous group of prokaryotes able to lyse and feed on the cellular constituents of other bacteria in conditions of nutrient scarcity. In this study, we describe the isolation of Actinobacteria predator of other bacteria from the marine water of the Moroccan Atlantic coast. Only 4 Actinobacteria isolates showing strong predation capability against native or multidrug-resistant Gram-positive or Gram-negative bacteria were identified among 142 isolated potential predatory bacteria. These actinobacterial predators were shown to belong to the Streptomyces genus and to inhibit the growth of various native or multidrug-resistant micro-organisms, including Micrococcus luteus, Staphylococcus aureus (native and methicillin-resistant), and Escherichia coli (native and ampicillin-resistant). Even if no clear correlation could be established between the antibacterial activities of the selected predator Actinobacteria and their predatory activity, we cannot exclude that some specific bio-active secondary metabolites were produced in this context and contributed to the killing and lysis of the bacteria. Indeed, the co-cultivation of Actinobacteria with other bacteria is known to lead to the production of compounds that are not produced in monoculture. Furthermore, the production of specific antibiotics is linked to the composition of the growth media that, in our co-culture conditions, exclusively consisted of the components of the prey living cells. Interestingly, our strategy led to the isolation of bacteria with interesting inhibitory activity against methicillin-resistant S. aureus (MRSA) as well as against Gram-negative bacteria.

Antimicrobial potential of actinomycetes isolated from the unexplored hot Merzouga desert and their taxonomic diversity.

The absence of new antibiotics is guiding more and more researchers to specific ecosystems. One hundred and sixty-three Actinobacteria isolates were isolated from Merzouga sand and screened for their antibacterial and antifungal activities. To test the antimicrobial effect of isolates, four microorganisms known as human potential pathogens were used. The electrophoretic profiles of isolates obtained by repetitive element PCR fingerprinting (rep-PCR) were compared by clustering. Results showed that among the tested isolates, 59% were active against one or more in testing Gram-positive, Gram-negative and the yeast Candida albicans The importance of culture media for the activity expression was revealed. Comparative analysis of antimicrobial activity divided isolates into 15 groups. The comparison of the average diameters of inhibition zones using Minitab V.17 allowed subdivision of the 15 groups into 20 subgroups. Dendrograms derived from the BOXA1R-PCR fingerprints showed that 36 isolates were grouped in 16 clusters, containing from two to four isolates while 127 isolates were not grouped. The tested antimicrobial activities showed a high biological diversity with important inhibition of pathogens tested. The rep-PCR revealed a high taxonomic diversity of isolates. The combination of antimicrobial activity and rep-PCR results revealed the diverse pattern of Merzouga sand dune Actinobacteria.

From Isolation of Phosphate Solubilizing Microbes to Their Formulation and Use as Biofertilizers: Status and Needs.

The production of biofertilizers at industrial level is a bottleneck because bacterial strains are generally developed and managed by research laboratories and not by production units. A seamless transition from laboratory to field application is, therefore necessary. This review provides an overview of the constraints that limiting the application or the implementation of Actinobacteria based biofertilizers especially in agricultural field and suggests solutions to overcome some of these limits. General processes of making and controlling the quality of the inoculum are briefly described. In addition, the paper underlines the opportunity of biofertilizers alone or in combination with chemical fertilizers. This review also, highlights the latest studies (until June 2019) and focuses on P-solubilization microorganisms mainly Actinobacteria. The biotechnology of these bacteria is a glimmer of hope for rock phosphate (RP) bioformulation. Since direct application of RP fertilizer is not always agronomically effective due to its sparse solubility.

Screening for Non-polyenic Antifungal Produced by Actinobacteria from Moroccan Habitats: Assessment of Antimycin A19 Production by Streptomyces albidoflavus AS25.

Fungal diseases are currently a serious public health problem, due to the limited number of fact-based effective principles, and the emergence of resistant strains to the polyenic antifungals. The aim of this study was to screen, for non-polyenic antifungals production by Actinobacteria, and to validate the screening program by characterizingthe produced compounds.Actinobacteria isolates were tested against four clinic human-pathogenic fungi isolated from Hospital Mohammed V Rabat, Morocco. The production of non-polyenic antifungal metabolites by active isolates was investigated based on the yeast cell specificity as challenging targets, antibacterial activity, activity against resistant Candida tropicalis R2 and Pythium irregular (resistant to polyenes), inhibition of antifungal activity by the addition of exogenous ergosterol, and the UV-visible light spectrophotometric analysis of the active crude extracts.The antifungal compound produced was purified using various chromatographic techniques and the selected producing strain was identified using the polyphasic approach.Among 480 Actinobacteria isolates, 55 showed antifungal activity against all tested clinically derived fungi. After performing the screening program, 4 Actinobacteria that hadall the desired criteriawere selected. Using the polyphasic approach, the taxonomic position of the selected Streptomyces AS25, isolated from rhizospheric soil of Alyssum spinosum, showed that it belongs to Streptomyces genus with 100% partial 16S similarity with Streptomyces albidoflavus NBRC13010. On the basis of HPLC and mass spectrometry, the purified compound produced by Streptomyces AS25 was identified as a non-polyenic lactone, antimycin A19, which has been found for the first time to be produced by Streptomyces albidoflavus strain. Following the obtained results, it is important to note that our screening criteria for non-polyenic antifungals have been validated and the rhizospheric soil represents an interesting source to isolate Actinobacteria.

Endophytic actinobacteria of medicinal plant Aloe vera: Isolation, antimicrobial, antioxidant, cytotoxicity assays and taxonomic study

Objective: To explore the new sources of novel bioactive compounds having pharmaceutical and agricultural interest and to search the endophytic actinobacteria from medicinal plants. Methods: NAF-1 an endophyte actinobacteria was isolated from leaves of medicinal plant Aloe vera collected in Marrakesh, Morocco using Bennett agar as selective medium. NAF-1 was tested for its antimicrobial activity against five pathogenic bacteria such asStaphylococcus aureus PIC 53156,Micrococcus luteus ATCC381,Bacillus subtilis ATCC 14579,Pseudomonas aeruginosa DSM 50090 and Escherichia coli ATCC 8739 and four human clinic fungi belonging to the Candida,Aspergillus and Microsporum genera. Several antioxidant activities were studied such as DPPH free radical scavenging,β-carotene and linoleic acid and reducing power assays. The total of phenol and flavonoid was also calculated. Using Artemia salina shrimp assay, the cytotoxicity of NAF-1 crude extract was determined.Results: The results revealed that the actinobacteria showed a high activity (≥20 mm) against only Gram positive bacteria but it had a moderate activity (between 13 and 15 mm) against Human clinic fungi. The isolate also exhibited a LD50 of 14.20 μg/mL in the cytotoxicity assay. The result showed that the crude extract presented an interesting free radical-scavenging activity with IC50 value of (5.58 ± 0.26) μg/mL and a high value of phenolic and flavonoid compounds with (15.41 ± 0.18) μg GAE/mg extract and (11.41± 0.06) μg QE/mg extract respectively. Moreover, the taxonomic position of our endophyte actinobacteria using the morphological and physiological criteria and using16SrRNA gene sequence (polyphasic approach) showed that the NAF-1 isolate was similar to Streptomyces hydrogenans which was never described as an endophyte actinobacteria. Conclusions: This isolated strain appears promising resources of bioactive agents and can be exploited to produce therapeutic agents active against pathogenic disease.

Artemia salina as a new index for assessment of acute cytotoxicity during co-composting of sewage sludge and lignocellulose waste.

Considering the necessity to constantly monitor the safety of use of sewage sludge, we have focused on evaluating the toxicity of raw sludge and sludge treated by co-composting with date palm waste using an in vitro assessment of cytotoxicity based on Artemia salina larvae as a simple new sensitive and reliable routine test. The efficiency of co-composting in decreasing sludge toxicity was evaluated in terms of cytotoxicity abatement reaching 100% by the second month of composting for mixture A (1/3 sludge+2/3 date palm waste) and the third month for mixture B (1/2 sludge+1/2 date palm waste). Cytotoxicity abatement was confirmed by the increase of germination index, which reached over 100% with positive correlation for lettuce (R(2)=0.81 and 0.86) and for turnip (R(2)=0.87 and 0.74) for mixtures A and B respectively. A strong correlation between the proposed cytotoxicity test and the evolution of regulatory physical-chemical approaches was found, (R(2)=0.88 and 0.89) for NH4(+)/NO3(-) and (R(2)=0.80 and 0.88) for C/N respectively for mixture A and B. These findings allow the inexpensive bioassay reported to be used as a highly sensitive test to determine the cytotoxicity and maturity of composts.