Micromonospora species from rarely-exploited Egyptian habitats: chemical profile, antimicrobial, and antitumor activities through antioxidant property.

The development of new anticancer agents with a selective action mechanism has become a significant scientific challenge, especially as cancers remain the world's leading cause of death. Actinobacteria and its bioactive compounds have recently become a promising perspective alternative to cancer therapy. In this study, some extracted metabolites of Micromonospora exhibited potent antimicrobial with microbial inhibition zone ≥ 7 mm, and cytotoxic activities against MCF-7 and HepG2 cell lines with promising activities ≥ 85%. Additionally, treatment of DENA/CCl4 rats with the strain Micromonospora sp1 has induced a substantial amelioration of the liver functions, enhancing liver architecture near normal and antioxidant properties through elevation of antioxidant enzyme levels. So that these preliminary results can provide metabolites from Micromonospora sp1 as an anti-liver cancer therapy. Finally, we introduced the chemical profiling of Micromonospora sp1 metabolic extract by LC-QTOF-MS-MS technique, where eight compounds with reported antioxidant property anti-liver cancer activity were targeted, validated as iNOS inhibitor through molecular docking studies. The findings in this study can be a significant step towards studying natural bioactive products produced by Micromonospora spp. as agents for anti-liver cancer. KEY POINTS: • Metabolites of Micromonospora strain from unexploited Egyptian habitats were investigated with LC/MS library-based chemical profile and molecular docking studies as iNOS inhibitors. • Some Micromonospora strains exhibited potent antimicrobial with microbial inhibition zone ≥ 7 mm, and cytotoxic activities against MCF-7 and HepG2 cell lines with promising activities ≥ 85%. • Micromonospora extract exhibited anti-liver cancer activity in vivo through the antioxidant property by inhibiting the liver cancer biomarkers (LDH and AFP) and enhancing other biochemical parameters.

Endophytic actinobacteria from wild medicinal plants are a natural source of insecticide to control the African cotton leafworm (Spodoptera littoralis).

Insecticide resistance in agricultural pests has prompted the need to discover novel compounds with new modes of action. We investigated the potency of secondary metabolites from seventy endophytic actinobacteria against laboratory and field strains of Spodoptera littoralis (fourth instar), comparable to the bioinsecticide spinetoram (Radiant SC 12%). Endophytes from Artemisia herba-alba and A. judaica were highly effective. Chemical profiling of the most potent metabolite of the strain Streptomyces sp. ES2 was investigated using LC-QTOF-MS-MS technique, and the activity was validated through molecular docking studies. Metabolic extracts from actinobacteria belonging to Streptomyces, Nocardioides, and Pseudonocardia showed immediate and latent death to the Spodoptera littoralis fourth instar larvae. The metabolite from strain ES2 has shown the most promising and significant histopathological and inhibitory effects on the fourth instar larvae. ES2 metabolite caused lesions in the body wall cuticle, indicating a different mode of action than that of Radiant. Chemical profiling of ES2 showed the presence of cyromazine (molt inhibitor), 4-nitrophenol, and diazinon as key constituents. In conclusion, these findings suggest that secondary metabolites from endophytic actinobacteria inhabiting wild medicinal plants can be a sustainable source for promising natural biocontrol agents. This is the first illustration of the insecticidal activity of Artemisia spp. microbiome, and natural cyromazine synthesis by actinobacteria.

Bioactivities of endophytic actinobacteria inhabiting Artemisia herba-alba emphasizing differences from free-living strains.

The endophytic actinobacteria associated with Artemisia herba-alba (synonym: Seriphidium herba-alba) are highly diverse. This study aimed to illustrate the extent of their differences from the free-living actinobacteria in the surrounding environment. A selection of eighteen actinobacteria inhabiting A. herba-alba were compared with twenty and ten actinobatceria isolates from the surrounding desert and groundwater, respectively, representing six genera. Antagonistic and enzymatic activities, plant growth-promoting traits, and the occurrence of biosynthetic genes were compared among the isolates. Data were analyzed statistically using principal component analysis (PCA) and were visualized using heat map. Endophytic strains showed higher antimicrobial activity and production of plant growth promoters compared to desert and groundwater strains. Polyketide synthase and non-ribosomal peptide synthetase gene clusters were detected at higher frequencies in the endophytic strains (8 and 11 strains, respectively) than the desert strains (1 and 2 strains, respectively). In contrast, both gene clusters were not detected in the groundwater strains. The PCA revealed unique metabolic characteristics of the endophytes. The heatmap clustered the endophytic strains apart from the free-living strains, indicating distinctive qualitative and quantitative bioactivities. Analysis of 16S rRNA genes confirmed the chemotaxonomic identity of all but two strains, with > 94.5% similarity. Six endophytes displayed < 99.5% similarity with their closest type strains, which might indicate species novelty. This study provides an evidence of functional differences and possible species novelty of the endophytic actinobacteria inhabiting A. herba-alba, compared with the free-living species.

Biocontrol of chocolate spot disease (Botrytis cinerea) in faba bean using endophytic actinomycetes Streptomyces: a field study to compare application techniques.

Sustainable agriculture is needing economic applications for disease control. One possibility is offered by local medical plants. Endophytes of medical plants, such as actinomycetes Streptomyces sp. have previously shown antagonistic activities against fungal phytopathogens. In the present field experiment, we aimed to verify the efficiency of endophytic Streptomyces against one of the common pathogens, Botrytis cinerea, causing chocolate spot disease for faba bean (Vicia fabae L.). We tested two strains of Streptomyces (MG788011, MG788012) and three techniques to apply the biocontrol agent: (1) coating the seeds with spores, (2) spraying mycelia and (3) spraying the crude metabolites over the plants. The technique using the crude metabolites was the most efficient to prevent the disease symptoms. Both of the endophytic strains diminished the disease symptoms and improved the plant growth. The study offers a potential biological control technique to prevent chocolate spot disease and, at the same time, increase the yields of faba bean in sustainable agriculture.

Prevalence of antibiotic resistant mastitis pathogens in dairy cows in Egypt and potential biological control agents produced from plant endophytic actinobacteria.

Dairy production is threatened by antibiotic resistant pathogens worldwide, and alternative solutions to treat mastitis are not available. The prevalence of antibiotic resistant strains is not well known in less developed countries. The prevalence of pathogenic bacteria and their resistance to 21 commercial antibiotics were studied in milk samples taken from 122 dairy cows suffering from the symptoms of mastitis in Egypt. The bacterial species were identified with molecular methods, and antibiotic resistance was studied with disc diffusion method. The prevalence of Streptococcus aureus, Escherichia coli and Pseudomonas aeruginosa were 30%, 17% and 3.5%, respectively. Most (90%) of the S. aureus strains showed resistance to penicillin whereas only 10% of the strains were resistant to oxacillin. Nearly half (40%) of E. coli strains showed resistance to streptomycin. Six P. aeruginosa strains showed resistance to several antibiotics, including ceftriaxone, enrofloxacin and levofloxacin. This points out that despite P. aeruginosa was not common, it should be followed up carefully. Potential biocontrol agents against antibiotic resistant mastitis bacteria were searched among 30 endophytic actinobacterial strains derived from wild medicinal plants. Three plants, namely Mentha longifolia, Malva parviflora and Pulicaria undulata were chosen for a more detailed study; their endophytic actinobacteria were used to prepare metabolic extracts. The crude metabolites of the actinobacteria were extracted with ethyl acetate. All metabolic extracts inhibited the growth of S. aureus, methicillin-resistant Staphylococcus aureus (MRSA), E. coli and P. aeruginosa in vitro. The 16S rRNA sequence analysis revealed that the most efficient actinobacterial strains were two Micromonospora sp. and one Actinobacteria bacterium. We conclude that the combination of the metabolites of several endophytic actinobacteria derived from several medicinal plants would be the most efficient against pathogens. Different metabolite cocktails should be studied further in order to develop novel biocontrol agents to treat antibiotic resistant mastitis bacteria in dairy cows.

An integrated bioaugmentation/electrocoagulation concept for olive mill wastewater management and the reuse in irrigation of biofuel plants: a pilot study.

A consortium of highly degrading microorganisms was used in an integrated bioaugmentation/electrocoagulation process for treating olive mill wastewater. The system was investigated for treating 1 m3 day-1, at a pilot scale, for 2 years; hydraulic loading rate and organic loading rate were 2880 l m-2 day-1 and 37,930 g COD m-2 day-1, respectively. Average removal efficiency for COD, oils, and total phenols was 63.9%, 85.2%, and 43.6%, respectively. The olive mill consortium, OMC, consisted of seven actinomycete strains. The strains were confirmed, by 16S rDNA analysis, to belong to five Streptomyces, one Kitasatospora, and one Micromonospora strains, at 100-99.06% similarities. Hydrolytic enzyme activities of OMC strains were remarkably higher for degrading cellulosic and lipid constituents (enzyme-cumulative indices, 14-16.1), than the phenolic constituents (indices, 4.1-6.5). The establishment of actinomycetes in the treatment system was indicated by their increased counts in the biofilm at the end of the biofilter, reaching 13-fold higher than that in the control bed. The treated effluent was toxic to the seedlings of Jatropha curcas (Jatropha) and Simmondsia chinensis (Jojoba). Though its application in irrigation of 3-year-old Jatropha shrubs, significantly, enhanced the fruit yield up to 1.85-fold higher than the control, without affecting the seed oil content, after 3-month application, the irrigated soil showed insignificant changes in its biochemical properties. This developed bioaugmentation/electrocoagulation process can treat wastewater with extremely high organic strength, while its approximate construction and operational costs are limited to 0.03 and 0.51 US$ m-3, respectively. It produces a treated effluent that can be reused in irrigation of specific plants. Graphical abstract.

Actinomycetes in rice straw decomposition.

Actinomycetes are well known as lignocellulose decomposers. Forty-two cellulolytic actinomycete isolates have been recovered from soil. Those were identified to the genus level as: Streptomyces (26 isolates), Nocardiopsis (5 isolates), Micromonospora (4 isolates), Nocardioides (4 isolates) in addition to three isolates tentatively identified as Nocardia, Kibdelosporangium and Saccharomonospora. These actinomycetes were examined for their cellulolytic activity using filter paper strips and rice straw pieces. Four isolates of the four genera Kibdelosporangium, Micromonospora, Streptomyces and Nocardioides were able to efficiently degrade rice straw pieces in minimal medium causing significant weight loss between 50 and 61%. Application of this knowledge may improve the management of waste rice straw.

Generic and functional diversity in endophytic actinomycetes from wild Compositae plant species at South Sinai - Egypt.

The diversity of culturable endophytic actinomycetes associated with wild Compositae plants is scantily explored. In this study, one hundred and thirty one endophytic actinobacteria were isolated from ten Compositae plant species collected from South Sinai in Egypt. Microscopic and chemotaxonomic investigation of the isolates indicated fourteen genera. Rare genera, such as Microtetraspora, and Intrasporangium, which have never been previously reported to be endophytic, were identified. Each plant species accommodated between three to eight genera of actinobacteria and unidentified strains were recovered from seven plant species. The generic diversity analysis of endophytic assemblages grouped the plant species into three main clusters, representing high, moderate and low endophytic diversity. The endophytes showed high functional diversity, based on forty four catabolic and plant growth promotion traits; providing some evidence that such traits could represent key criteria for successful residence of endophytes in the endosphere. Stress-tolerance traits were more predictive measure of functional diversity differences between the endophyte assemblages (Shannon's index, p = 0.01). The results indicate a potential prominent role of endophytes for their hosts and emphasize the potency of plant endosphere as a habitat for actinobacteria with promising future applications.