Indigenous case of mycetoma by Actinomadura madurae in Paraná (Entre Ríos, Argentina): case report and literature review of the argentinian casuistry

El micetoma es una infección granulomatosa crónica que involucra tejidos cutáneos, subcutáneos y eventualmente músculo y hueso. Puede ser causada por bacterias Gram positivas filamentosas (actinomicetomas) o por hongos (eumicetomas). Es una infección endémica de zonas subtropicales con baja humedad relativa. El objetivo de este trabajo es el de presentar un caso de actinomicetoma causado por Actinomadura madurae autóctono de la ciudad de Paraná (Entre Ríos) con el fin de demostrar que pueden darse casos esporádicos por fuera de las zonas endémicas descritas. Además, se realizó una revisión bibliográfica de los casos de micetoma descritos en Argentina y se los comparó con el caso reportado.Mycetoma is a chronic granulomatous infection involving skin, subcutaneous tissue and eventualy muscle and bone. It can be caused by Gram positive filamentous bacteria (actinomycetomas) or fungi (eumicetomas). It is an endemic infection in subtropical areas with low relative humidity. The aim of this work is to present a case of actinomycetoma caused by Actinomadura madurae in a patient from Paraná city (Entre Rios) to demonstrate that sporadic mycetoma cases may occur outside the described endemic areas. In addition, a literature review of Argentinian mycetoma cases was performed.Fil: Dudiuk, Catiana Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; Argentina. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas; ArgentinaFil: Theill, L.. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas; ArgentinaFil: Moyano, Susana. Laboratorio Privado Avenida. Paraná, Entre Ríos; ArgentinaFil: Barbagelata, María Sol. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas; ArgentinaFil: Leonardelli, Florencia. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Unidad de Administración Territorial; ArgentinaFil: Macedo, Daiana. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas; ArgentinaFil: Latorre Rapella, María Gabriela. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas; ArgentinaFil: Acosta, A.. Laboratorio Privado Avenida. Paraná, Entre Ríos; ArgentinaFil: Gamarra, S.. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas; ArgentinaFil: Garcia, Guillermo Manuel. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; Argentin

Actinomadura rubteroloni sp. nov. and Actinomadura macrotermitis sp. nov., isolated from the gut of the fungus growing-termite Macrotermes natalensis

The taxonomic positions of two novel aerobic, Gram-positive actinobacteria, designated strains RB29$^{T}$ and RB68T, were determined using a polyphasic approach. Based on 16S rRNA gene sequence analysis, the closest phylogenetic neighbours of RB29$^{T}$ were identified as Actinomadura rayongensis DSM 102126$^{T}$ (99.2 % similarity) and Actinomadura atramentaria DSM 43919$^{T}$ (98.7 %), and for strain RB68$^{T}$ was Actinomadura hibisca DSM 44148$^{T}$ (98.3 %). Digital DNA–DNA hybridization (dDDH) between RB29$^{T}$ and its closest phylogenetic neighbours, A. rayongensis DSM 102126$^{T}$ and A. atramentaria DSM 43919$^{T}$, resulted in similarity values of 53.2 % (50.6–55.9 %) and 26.4 % (24.1–28.9 %), respectively. Additionally, the average nucleotide identity (ANI) was 93.2 % (94.0 %) for A. rayongensis DSM 102126$^{T}$ and 82.3 % (78.9 %) for A. atramentaria DSM 43919$^{T}$. dDDH analysis between strain RB68$^{T}$ and A. hibisca DSM 44148$^{T}$ gave a similarity value of 24.5 % (22.2–27.0 %). Both strains, RB29$^{T}$ and RB68$^{T}$, revealed morphological characteristics and chemotaxonomic features typical for the genus Actinomadura, such as the presence of meso-diaminopimelic acid in the cell wall, galactose and glucose as major sugar components within whole-cell hydrolysates and the absence of mycolic acids. The major phospholipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol and phosphatidylinositol mannoside. Predominant menaquinones were MK-9(H$_{6}$) and MK-9(H$_{8}$) for RB29$^{T}$ and MK-9(H$_{4}$) and MK-9(H$_{6}$) for RB68T. The main fatty acids were identified as 10-methyloctadecanoic acid (10-methyl C$_{18:0}$), 14-methylpentadecanoic acid (iso-C$_{16:0}$), hexadecanoic acid (C$_{16:0}$) and cis-9-octadecanoic acid (C$_{18:1}$ ω9c). Here, we propose two novel species of the genus Actinomadura: Actinomadura rubteroloni sp. nov. with the type strain RB29$^{T}$ (=CCUG 72668$^{T}$=NRRL B-65537$^{T}$) and Actinomadura macrotermitis sp. nov. with the type strain RB68$^{T}$ (=CCUG 72669$^{T}$=NRRL B-65538$^{T}$)

Actinomadura meyerae osteitis following wound contamination with hay in a woman in France: a case report

<p>Abstract</p> <p>Introduction</p> <p>Mycetoma is a chronic granulomatous infection caused by environmental fungi or bacteria. It affects dermal and subcutaneous tissues, with putative contiguous extension to muscles or bones. While common in tropical and subtropical areas, mycetoma is rare in Europe.</p> <p>Case presentation</p> <p>We describe a case of <it>Actinomadura meyerae </it>osteitis in a 49-year-old Caucasian woman who suffered a tibia open fracture contaminated with hay; to the best of our knowledge the first case of autochthonous <it>A. meyerae </it>infection reported in France. The bacterium was cultivated from a bone biopsy. Following surgical osteosynthesis and six months of treatment with cotrimoxazole, our patient made a full recovery.</p> <p>Conclusion</p> <p>Our case report suggests that <it>A. meyerae </it>is a potential agent of wound infection in farm workers in contact with hay.</p

Identification of a new strain of Actinomadura isolated from Saharan soil and partial characterization of its antifungal compounds

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

Taxonomy of mycelial actinobacteria isolated from Saharan soils and their efficiency to reduce aflatoxin B1 content in a solid-based medium

Aflatoxin B1 (AFB1) is a carcinogenic compound produced by filamentous fungi. In order to reduce AFB1 occurrence in foodstuffs, 13 strains of mycelial actinobacteria were tested in vitro for the efficacy to reduce AFB1 content; all were isolated from the Saharan soils of Algeria. Firstly, morphological study and molecular analysis, based on the 16S rRNA gene, indicated that these strains belong to Actinomadura, Nocardiopsis, Nonomuraea, Saccharothrix and Streptomyces genera. Secondly, each strain’s efficacy to reduce pure AFB1 content was studied in ISP2-medium. After a 4-day incubation at 30°C on AFB1-supplemented medium (5 ppm of AFB1), AFB1 was extracted and quantified. AFB1 content was reduced by all strains (42.9–97.6%). The three most efficient reducers (94.9–97.6%) were two strains belonging to the genus Streptomyces and one to the genus Saccharothrix. Among the latter, strains ACD6 and ABH19 showed no adsorption mechanism involved, suggesting a potential degradation mechanism. These findings led us to suggest that these actinobacterial strains could be used as decontamination treatments for the reduction of AFB1 content

Actinomadura adrarensis sp. nov., an actinobacterium isolated from Saharan soil

A novel actinobacterial strain, designated ACD12T, was isolated from a Saharan soil sample collected from Adrar province, southern Algeria. A polyphasic study was carried out to establish the taxonomic position of this strain. Strain ACD12T was observed to form extensively branched substrate mycelia. Aerial mycelium was absent or was weakly produced on all media tested, while spore chains were short with a hooked and irregular spiral form (2–3 turns). The dominant diaminopimelic acid isomer in the cell wall was meso-diaminopimelic acid. Glucose, ribose, galactose, mannose and madurose occured in whole-cell hydrolysates. The major phospholipid was diphosphatidylglycerol and phosphatidylinositol. The predominant menaquinone was MK-9(H6). The fatty acid profile was characterized by the presence of C16 : 0, C17 : 0, C15 : 0, C18 : 0, C18 : 1 cis9 and iso-C16 : 0. Results of 16S rRNA gene sequence comparisons revealed that strain ACD12T shared the highest degree of 16S rRNA gene sequence similarity with Actinomadura sputi DSM 45233T (98.3 %) and Actinomadura hallensis DSM 45043T (97.8 %). All tree-making algorithms used also supported strain ACD12T forming a distinct clade with its most closely related species. In addition, DNA–DNA hybridization indicated only 39.8 % relatedness with A. sputi DSM 45233T and 18.7 % relatedness with A. hallensis DSM 45043T. The combined phenotypic and genotypic data show that the novel isolate represents a novel species of the genus Actinomadura , for which the name Actinomadura adrarensis sp. nov., is proposed, with the type strain ACD12T (=DSM 46745T =CECT 8842T)

Isolamento e bioprospecção de actinomicetos endofíticos de Vochysia divergens Pohl

Orientadora : Profª Drª Chirlei GlienkeCoorientadora : Profª Drª Daiani C. SaviDissertação (mestrado) - Universidade Federal do Paraná, Setor de Ciências Biológicas, Programa de Pós-Graduação em Ciencias Biológicas (Microbiologia, Parasitologia e Patologia Básica). Defesa: Curitiba, 27/04/2017Inclui referências : f. 71-77Resumo: O Pantanal é uma planície de inundação periódica detentora de uma biodiversidade singular, e possui diversas plantas com conhecimento tradicional associado às suas propriedades medicinais. Entre estas, a planta Vochysia divergens, conhecida como Cambará, é utilizada em tratamentos de resfriados, tosse, febre e pneumonia, e se mostra promissora para a prospecção de microrganismos. Dos quais os endófitos isolados de plantas medicinais têm sido associados a uma ampla gama de metabólitos secundários com atividade biológica. Neste contexto, o presente trabalho objetivou o isolamento e a bioprospecção de actinomicetos endofíticos da planta medicinal V. divergens. Um total de 10 isolados obtidos foram identificados por meio do sequenciamento parcial do gene 16S rRNA e análise morfológica, como Aeromicrobium ponti, Williamsia serinedens, Streptomyces thermocarboxydus, Microbacterium sp., Sphaerisporangium sp., Micrococcus sp., Microbispora sp. e Actinomadura sp.. Afim de selecionar a melhor condição de cultivo para produção de metabólitos secundários, foram avaliadas duas condições diferentes de meio de cultura e temperatura. Todas as condições utilizadas resultaram na produção de metabólitos com atividade biológica, porém os melhores resultados foram no cultivo em caldo SG (Soja e glucose) a 36°C. Nestas condições, o extrato a partir da fermentação da linhagem Aeromicrobium ponti (LGMB491) apresentou atividade contra Staphylococcus aureus sensível à meticilina (MSSA) e S. aureus resistente à meticilina (MRSA), com CIM de 0,02mg/mL e 0,04mg/mL, respectivamente. O extrato oriundo da linhagem Microbacterium sp. (LGMB471) apresentou inibição total do crescimento micelial do fitopatógeno Phyllosticta citricarpa com CIM de 0,16mg/mL. Neste estudo, o conhecimento da comunidade endofítica da planta medicinal V. divergens foi ampliado por meio do isolamento de novos gêneros associados a esta planta, alguns deles isolados pela primeira vez como endófitos. Palavras chaves: Vochysia divergens. Pantanal. Actinomicetos endofíticos. MRSA.Abstract: The Pantanal is a periodic flood plain with a unique biodiversity, and has several plants with traditional knowledge associated with its medicinal properties. The plant Vochysia divergens, known as Cambará, is used for treatment of colds, cough, fever and pneumonia, and is very promising for microorganisms' prospection. Endophytic microorganisms isolated from medicinal plants have been studied due to the production of a wide range of secondary metabolites with biological activity. In this context, the present work aimed at the isolation and bioprospection of endophytic actinomycetes from medicinal plant V. divergens. Ten strains of endophytic actinomycetes were identified by partial 16S rRNA gene sequencing and morphological analysis, as belonging to Aeromicrobium ponti, Williamsia serinedens, Streptomyces thermocarboxydus, Microbacterium sp., Sphaerisporangium sp., Micrococcus sp., Microbispora sp. and Actinomadura sp. In order to select the best culture conditions for production of secondary metabolites, two different culture media and two temperatures were evaluated. All the conditions used resulted in the production of metabolites with biological activity, but the best conditions are cultivation in SG broth at 36°C. In these conditions, the extract from the fermentation of Aeromicrobium ponti (LGMB491) showed high activity against methicillin-sensitive Staphylococcus aureus (MSSA) and methicillin-resistant S. aureus (MRSA), with MIC of 0.02mg/mL and 0.04mg/mL, respectively. The extract from the Microbacterium sp. (LGMB471) showed total inhibition of the mycelial growth of phytopathogen Phyllosticta citricarpa with MIC of 0.16mg/mL. In this study, we broadened the knowledge about the endophytic community of the medicinal plant V. divergens through the isolation of rare actinomycetes, some of them isolated for the first time as endophytes. Key-words: Vochysia divergens. Pantanal. Endophitics actinomycetes. MRSA

Streptomyces smaragdinus sp. nov., isolated from the gut of the fungus growing-termite Macrotermes natalensis

The taxonomic position of a novel aerobic, Grampositive actinobacteria, designated strain RB5(T), was determined using a poly phasic approach. The strain, isolated from the gut of the fungusfarming termite Macrotermes natalensis, showed morphological, physiological and chemotaxonomic properties typical of the genus Streptomyces. Based on 16S rRNA gene sequence analysis, the closest phylogenetic neighbour of RB5(T) was Streptomyces polyrhachis DSM 42102(T) (98.87 %). DNA-DNA hybridization experiments between strain RB5(T) and S. polyrhachis DSM 42102(T) resulted in a value of 27.4 % (26.8 %). The cell wall of strain RB5(T) contained iota iota diaminopimelic acid as the diagnostic amino acid. Mycolic acids and diagnostic sugars in whole cell hydrolysates were not detected. The strain produced the following major phospholipids: diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol, phosphatidylinositolmannoside and phosphatidylserine. The menaquinone profile showed hexaand octahydrogenated menaquinones containing nine isoprene units [MK-9(H-6) and MK-9(H-8)]. The strain exhibited a fatty acid profile containing the following major fatty acids: 12methyltridecanoic acid (isoC(14:0)) 12-methyltetradecanoic acid (anteiso-C-15:0), 13-methyltetradecanoic acid (isoC(15:0)) and 14-methylpentadecanoic acid (isoC(16:0)). Here, we propose a novel species of the genus Streptomyces - Streptomyces smaragdinus with the type strain RB5(T) (=VKM Ac-2839(T)=NRRL B65539(T))

Novel actinobacterial diversity in arid Atacama Desert soils as a source of new drug leads

PhD ThesisThe search for new specialised metabolites, notably antibiotics, that can be developed for healthcare has steadily shifted towards the isolation and screening of rare and novel actinobacteria from extreme habitats on the premise that such habitats give rise to unique biodiversity that is the basis of novel chemistry. To this end, a taxonomic approach to bioprospecting for bioactive compounds was used to selectively isolate, dereplicate and classify actinobacteria from hyper-arid and extreme hyper-arid areas of the Atacama Desert in northwest Chile, namely from the Salar de Atacama and Yungay regions, respectively. Sample pretreatment and selective isolation strategies enabled the recovery of actinobacteria from each of these habitats and while population sizes were small, taxonomic diversity was high. Relatively large numbers of Amycolatopsis and Streptomyces strains were isolated from the hyper-arid Salar de Atacama soil, as were smaller numbers of Actinomadura, Kribbella, Lechevalieria, Nonomuraea and Saccharothrix strains. In contrast, Modestobacter and Streptomyces isolates predominated in the extreme hyper-arid Yungay soil, the latter also contained smaller numbers of Blastococcus, Couchioplanes, Geodermatophilus and Pseudonocardia strains. With few exceptions representatives of these genera formed distinct phyletic lines in 16S rRNA gene trees. Polyphasic studies carried out on strains of ecological and biotechnological interest showed that isolates assigned to the genera Modestobacter and Streptomyces belonged to putative new species, as exemplified by the proposal for Streptomyces leeuwenhoekii sp. nov. for strains that formed a distinct branch in the Streptomyces 16S rRNA gene tree. In contrast, representatives of the genus Amycolatopsis were assigned to known species, albeit ones classified in a rare taxon, the Amycolatopsis 16S rRNA gene clade. Most of the representative isolates examined in standard plug assays inhibited the growth of one or more of a panel of five wild type microorganisms. In addition, some of the representative streptomycetes from the hyper-arid Salar de Atacama soil were found to inhibit cell envelope, cell wall, fatty acid and RNA synthesis in assays based on the use of Bacillus subtilis reporter genes. The results of this project demonstrate for the first time that hyper-arid and extreme hyper-arid Atacama Desert soils are rich reservoirs of cultivable rare and novel actinobacteria with the capacity to produce a broad range of bioactive compounds that can be developed as drug leads for medicine. Indeed, microorganisms, unlike plants and animals, have overcome the prevailing harsh conditions of the Desert. Life abounds in the Atacama Desert, but most of it is microbial!Royal Thai Government, and from the Thailand Institute of Scientific and Technological Research (TISTR

Adaptation to endophytic lifestyle through genome reduction by Kitasatospora sp. SUK42

Endophytic actinobacteria offer great potential as a source of novel bioactive compounds. In order to investigate the potential for the production of secondary metabolites by endophytes, we recovered a filamentous microorgansism from the tree Antidesma neurocarpum Miq. After phenotypic analysis and whole genome sequencing we demonstrated that this organism, SUK42 was a member of the actinobacterial genus Kitasatospora. This strain has a small genome in comparison with other type strains of this genus and has lost metabolic pathways associated with Stress Response, Nitrogen Metabolism and Secondary Metabolism. Despite this SUK42 can grow well in a laboratory environment and encodes a core genome that is consistent with other members of the genus. Finally, in contrast to other members of Kitasatospora, SUK42 encodes saccharide secondary metabolite biosynthetic gene clusters, one of which with similarity to the acarviostatin cluster, the product of which displays α-amylase inhibitory activity. As extracts of the host plant demonstrate this inhibitory activity, it suggests that the potential medicinal properties of A. neurocarpum Miq might be provided by the endophytic partner and illustrate the potential for exploitation of endophytes for clinical or industrial uses