Characterization of anti-BCG benz[α]anthraquinones and new siderophores from a Xinjiang desert-isolated rare actinomycete Nocardia sp. XJ31.

The current global demand for novel anti-TB drugs has drawn urgent attention on the discovery of natural product compounds with anti-TB activity. Lots of efforts have emphasized on environmental samples from unexplored or underexplored natural habits and identified numerous rare actinomycete taxa producing structurally diverse bioactive natural products. Herein, we report a survey of the rare actinobacteria diversity in Xinjiang region together with the discovery of anti-TB active natural products from these strains. We have collected 17 soil samples at different sites with different environmental conditions, from which 39 rare actinobacteria were identified by using a selective isolation strategy with 5 media variations. Among those isolated strains, XJ31 was identified as a new Nocardia sp. based on 16S rRNA gene analysis. Through one strain-many compounds (OSMAC) strategy combined with anti-Bacillus Calmette-Guérin bioassay-guided isolation, two groups of compounds were identified. They were twelve siderophores (nocardimicins, 1-12) and two anthraquinones (brasiliquinones, 13 and 14) and ten of them were identified as new compounds. The structures of the purified compounds were elucidated using HR-ESI-MS, 1D NMR, and 2D NMR techniques. The anti-TB bioassays revealed that the two benz[α]anthraquinones have potent activity against BCG (MICs = 25 µM), which can be used as a promising start point for further anti-TB drug development. KEY POINTS: • Ten new natural products were identified from Nocardia sp. XJ31. • Brasiliquinones 13 and 14 showed moderate anti-BCG activity.

Production of a Novel Tetrahydroxynaphthalene (THN) Derivative from Nocardia sp. CS682 by Metabolic Engineering and Its Bioactivities.

Nargenicin A1 is major secondary metabolite produced by Nocardia sp. CS682, with an effective antibacterial activity against various Gram-positive bacteria. Most Nocardia spp. have metabolic ability to produce compounds of diverse nature, so one-strain-many-compounds (OSMAC) approach can be applied for obtaining versatile compounds from these strains. In this study, we characterized a novel 1, 3, 6, 8-tetrahydroxynaphthalene (THN) derivative by metabolic engineering approach leading to the inactivation of nargenicin A1 biosynthesis. By using genome mining, metabolite profiling, and bioinformatics, the biosynthetic gene cluster and biosynthetic mechanism were elucidated. Further, the antibacterial, anticancer, melanin formation, and UV protective properties for isolated THN compound were performed. The compound did not exhibit significant antibacterial and cytotoxic activities, but it exhibited promising UV protection effects. Thus, metabolic engineering is an effective strategy for discovering novel bioactive molecules.

Cytotoxic activity of actinomycetes Nocardia sp. and Nocardiopsis sp. associated with marine sponge Amphimedon sp.

Cancer is a hazard life-threatening disease, which affect huge population worldwide. Marine actinomycetes are considered as promising source for potential chemotherapeutic agents. In our study, we carried out metabolic profiling for Nocardia sp. UR 86 and Nocardiopsis sp. UR 92 that were cultivated from the Red Sea sponge Amphimedon sp. to investigate their chemical diversity using different media conditions. The crude culture extracts were subjected to high-resolution mass spectrometry (HRMS) analysis. The chemical profiles of the different extracts of Nocardia sp. UR 86 and Nocardiopsis sp. UR 92 revealed their richness in diverse metabolites and consequently twenty compounds (1-20) were annotated. Moreover, the obtained extracts of the differently cultivated Nocardia sp. UR 86 and Nocardiopsis sp. UR 92 were investigated against three cell lines HepG2, MCF-7 and CACO2 and revealed the targeted cytotoxicity of Nocardia sp. and Nocardiopsis sp. metabolites against the three cell lines.

Poly-cis-isoprene Degradation by Nocardia sp. BSTN01 Isolated from Industrial Waste.

The natural and synthetic rubber (NR and SR) products are made up of poly-cis-isoprene which are estimated as one of the major solid-wastes and need to be cleared through bacterial bioremediation. The present research reports isolation and characterization of a gram-positive, non-spore forming, filamentous actinomycete Nocardia sp. BSTN01 from the waste of a rubber processing industry. We found NR- and SR-dependent growth of BSTN01 over a period of time. BSTN01 has been found to degrade NR by 55.3% and SR by 45.9% in 6 weeks. We have found an increase in the total protein of BSTN01 cells up to 623.6 and 573.9 µg/ml for NR and SR, respectively, after 6 weeks of growth in rubber-supplemented MSM medium. Scanning electron microscopy revealed adhesive growth of BSTN01 on the surface of NR and SR. Formation of aldehyde groups due to the degradation was indicated by Schiff's test and confirmed by FTIR-ATR analysis. The genome sequence of BSTN01 revealed the gene responsible for rubber degradation. The presence of lcp gene and structural analysis of the latex clearing protein further confirmed the reliability. Studies on quantification of rubber degradation capability by the isolated strain prove it to be an efficient degrader of NR and SR. This study revealed the genome sequence and structural analysis of the proteins responsible for degradation of rubber. A new fast-growing Nocardia strain can degrade both NR and SR with higher efficiency and have future potential for rubber solid-waste management either alone or in consortia.

Lichen-associated bacteria transform antibacterial usnic acid to products of lower antibiotic activity.

Lichens are specific symbiotic organisms harboring various microorganisms in addition to the two classic partners (algae or cyanobacterium and fungus). Although lichens produce many antibiotic compounds such as (+)-usnic acid, their associated microorganisms possess the ability to colonize an environment where antibiosis exists. Here, we have studied the behavior of several lichen-associated bacterial strains in the presence of (+)-usnic acid, a known antibiotic lichen compound. The effect of this compound was firstly evaluated on the growth and metabolism of three bacteria, thus showing its ability to inhibit Gram-positive bacteria. This inhibition was not thwarted with the usnic acid producer strain Streptomyces cyaneofuscatus. The biotransformation of this lichen metabolite was also studied. An ethanolamine derivative of (+)-usnic acid with low antibiotic activity was highlighted with chemical profiling, using HPLC-UV combined with low resolution mass spectrometry. These findings highlight the way in which some strains develop resistance mechanisms. A methylated derivative of (+)-usnic acid was annotated using the molecular networking method, thus showing the interest of this computer-based approach in biotransformation studies.

Broad Spectrum Antimicrobial Activity of Forest-Derived Soil Actinomycete, Nocardia sp. PB-52.

A mesophilic actinomycete strain designated as PB-52 was isolated from soil samples of Pobitora Wildlife Sanctuary of Assam, India. Based on phenotypic and molecular characteristics, the strain was identified as Nocardia sp. which shares 99.7% sequence similarity with Nocardia niigatensis IFM 0330 (NR_112195). The strain is a Gram-positive filamentous bacterium with rugose spore surface which exhibited a wide range of antimicrobial activity against Gram-positive bacteria including methicillin-resistant Staphylococcus aureus (MRSA), Gram-negative bacteria, and yeasts. Optimization for the growth and antimicrobial activity of the strain PB-52 was carried out in batch culture under shaking condition. The optimum growth and antimicrobial potential of the strain were recorded in GLM medium at 28°C, initial pH 7.4 of the medium and incubation period of 8 days. Based on polyketide synthases (PKS) and non-ribosomal peptide synthetases (NRPS) gene-targeted PCR amplification, the occurrence of both of these biosynthetic pathways was detected which might be involved in the production of antimicrobial compounds in PB-52. Extract of the fermented broth culture of PB-52 was prepared with organic solvent extraction method using ethyl acetate. The ethyl acetate extract of PB-52 (EA-PB-52) showed lowest minimum inhibitory concentration (MIC) against S. aureus MTCC 96 (0.975 µg/mL) whereas highest was recorded against Klebsiella pneumoniae ATCC 13883 (62.5 µg/mL). Scanning electron microscopy (SEM) revealed that treatment of the test microorganisms with EA-PB-52 destroyed the targeted cells with prominent loss of cell shape and integrity. In order to determine the constituents responsible for its antimicrobial activity, EA-PB-52 was subjected to chemical analysis using gas chromatography-mass spectrometry (GC-MS). GC-MS analysis showed the presence of twelve different chemical constituents in the extract, some of which are reported to possess diverse biological activity. These results confirmed that the presence of bioactive constituents in EA-PB-52 could be a promising source for the development of potent antimicrobial agents effective against wide range of microbial pathogens including MRSA.

A new natural nucleotide and other antibacterial metabolites from an endophytic Nocardia sp.

Nine compounds were isolated from Nocardia sp. YIM 64630, and their structures were elucidated as 5'-O-acetyl-2'-deoxyuridine (1), 22E,24R-5α,6α-epoxyergosta-8(14),22-diene-3ß,7α-diol (2), 22E,24R-5α,6α-epoxyergosta-8,22-diene-3ß,7α-diol (3), 22E,24R-ergosta-7,22-diene-3ß,5α,6ß-triol (4), 5α,8α-epidioxyergosta-6,22-dien-3ß-ol (5), 4',5,6-trihydroxy-7-methoxyisoflavone (6), 2,4,4'-trihydroxy-deoxybenzoin (7), methyl [4-hydroxyphenyl]acetate (8) and daidzein by extensive spectroscopic analyses. Compound 1 was isolated from natural resources for the first time. The antimicrobial and antioxidant activities of compounds 1-8 were investigated.

Características epidemiológicas, clínico-patológicas e morfotintoriais de quatorze casos de nocardiose em cães / Epidemiological, clinical, pathological and morphotinctorial features of fourteen cases of nocardiosis in dogs

Este artigo descreve os aspectos epidemiológicos, clínicos, patológicos e características morfotintoriais em quatorze casos de nocardiose em cães. Para isso foi realizado um estudo retrospectivo durante o período de janeiro de 2005 a dezembro de 2015 e selecionados os casos sugestivos de nocardiose. A identificação e caracterização do agente foi realizada através de técnicas histoquímicas especiais de Metenamina nitrato de prata de Grocott (GMS), Ziehl-Neelsen modificado, coloração de Gram do tipo Brown-Brenn modificado e Giemsa. Foram afetados predominantemente filhotes e em doze casos havia associação com o vírus da cinomose canina (VCC). Os sinais clínicos variaram de alterações respiratórias, neurológicas e cutâneas, relacionadas principalmente à infecção concomitante pelo VCC. Macroscopicamente haviam áreas multifocais a coalescentes, branco-amareladas, firmes, elevadas na superfície e que se aprofundavam ao corte, por vezes com material purulento, entremeadas por áreas avermelhadas irregulares afetando principalmente pulmões, linfonodos, fígado, rins e encéfalo. As lesões cutâneas foram observadas predominantemente nas regiões cervical e inguinal e variaram de supurativas a piogranulomatosas. No exame histopatológico a lesão era caracterizada principalmente por inflamação piogranulomatosa, porém em algumas áreas havia predomínio de inflamação necrossupurativa, e frequentemente eram observados macrófagos epitelioides, formando aglomerados de forma radiada, muitas vezes lembrando pseudorosetas. Nas técnicas histoquímicas foram observadas estruturas filamentosas, ramificadas, não septadas, medindo aproximadamente 1µm de espessura, impregnadas na coloração de prata, coradas em vermelho no Ziehl-Neelsen modificado, em azul na coloração de Gram do tipo Brown-Brenn modificado e fracamente rósea pálido no Giemsa. A nocardiose deve ser considerada em animais jovens que apresentam sinais respiratórios e neurológicos progressivos, bem como em lesões cutâneas com envolvimento do subcutâneo e linfonodos regionais. Deve ser investigado ainda um provável fator predisponente, como a infecção pelo vírus da cinomose canina e hemoparasitoses. O diagnóstico foi estabelecido através do exame histopatológico baseando-se na morfologia da bactéria e suas características histoquímicas, distinguindo principalmente de outros agentes bacterianos e fúngicos, constituindo uma importante ferramenta para o diagnóstico, quando não é possível a coleta do material para cultivo e isolamento do agente.(AU)

This paper describes the epidemiological, clinical, pathological and morphotinctorial characteristics in fourteen cases of nocardiosis in dogs. A retrospective study for the period of January 2005 to December 2015 was made and selected suggestive cases of nocardiosis. The identification and characterization of the agent was performed by special histochemical techniques Methenamine silver nitrate Grocott (GMS), modified Ziehl-Neelsen, Gram stain type modified Brown-Brenn and Giemsa. Were affected predominantly young and in twelve cases were associated with canine distemper virus (CDV). Clinical signs vary from respiratory, neurological and skin changes, mainly related to concomitant infection by CDV. Macroscopically had multifocal areas coalescing, yellowish-white, firm, elevated in surface and deepened the court, sometimes with purulent material, intermixed by irregular reddened areas affecting mainly lungs, lymph nodes, liver, kidneys, and brain. The cutaneous lesions were predominantly observed in cervical and inguinal and ranged from suppurative well as pyogranulomatous. In the histopathologic examination the injury was mainly characterized by inflammation pyogranulomatous, but in some areas there was a predominance of necrossupurativa inflammation, epithelioid macrophages and were frequently observed, forming clusters radiated form, often reminding rosettes. In the histochemical techniques were observed filamentous structures, branched, non-septate, measuring approximately 1µm thick, impregnated on silver staining, stained in red on the modified Ziehl-Neelsen, in blue on Gram stain type modified Brown-Brenn and weakly pale pink in Giemsa. The nocardiosis should be considered in young animals with progressive respiratory and neurological signs, as well as skin lesions involving the subcutaneous tissue and regional lymph nodes. It should be further investigated a possible predisposing factor, such as infection by canine distemper virus and hemoparasites. The diagnosis was established by histopathological examination based on the morphology of the bacteria and their histochemical characteristics, distinguishing mainly from other bacterial and fungal agents and is an important tool for the diagnosis, when the collection of material for cultivation and isolation of the agent is not possible.(AU)

Actinomicetoma torácico con afección pulmonar. Tratamiento con carbapenémico y aminoglucósido / Thoracic actinomycetoma with lung compromise. Treatment with carbapenem and aminoglycoside

Resumen: El actinomicetoma, infección crónica, granulomatosa y progresiva de la piel y tejido celular subcutáneo que afecta más las extremidades inferiores y en especial los pies, se caracteriza por la tríada de trayectos fistulosos, aumento de volumen y granos. La afección en el tórax en México ocupa el segundo lugar en cuanto a frecuencia. Sin embargo, el daño pulmonar es muy raro, afecta menos de 1% de los casos, éste puede ocurrir por continuidad al afectar el tronco, lo que causa tratamientos antimicrobianos prolongados con falta de apego a los mismos, así como aumento de la mortalidad. El objetivo de este artículo es comunicar un caso clínico de actinomicetoma pulmonar, así como la respuesta al tratamiento prescrito a este paciente con base en carbapenémicos, aminoglucósidos y trimetoprim con sulfametoxazol.

Abstract: Actinomycetoma is a chronic, granulomatous and progressive infection of the skin and subcutaneous cellular tissue that affects the lower limbs and especially the feet, characterized by triad of localized swelling, draining sinuses and grains. The chest condition in Mexico ranks second in frequency. Nevertheless, pulmonary affectation is very rare with less than 1%, this can occur by continuity affecting the trunk leading to prolonged antimicrobial therapy and lack of attachment to it, as well as increased mortality. The aim of this paper is to report a case of pulmonary actinomycetoma as well as response to treatment with carbapenems, aminoglycoside and trimethoprim with sulfamethoxazole.