Fyans, Joanna K; Bown, Luke; Bignell, Dawn R D
Potato common scab (CS) is an economically important crop disease that is caused by several members of the genus Streptomyces. In this study, we characterized the plant-pathogenic Streptomyces spp. associated with CS-infected potato tubers harvested in Newfoundland, Canada. A total of 17 pathogenic Streptomyces isolates were recovered from potato scab lesions, of which eight were determined to be most similar to the known CS pathogen S. europaeiscabiei. All eight S. europaeiscabiei isolates were found to produce the thaxtomin A phytotoxin and to harbor the nec1 virulence gene, and most also carry the putative virulence gene tomA. The remaining isolates appear to be novel pathogenic species that do not produce thaxtomin A, and only two of these isolates were determined to harbor the nec1 or tomA genes. Of the non-thaxtomin-producing isolates, strain 11-1-2 was shown to exhibit a severe pathogenic phenotype against different plant hosts and to produce a novel, secreted phytotoxic substance. This is the first report documenting the plant-pathogenic Streptomyces spp. associated with CS disease in Newfoundland. Furthermore, our findings provide further evidence that phytotoxins other than thaxtomin A may also contribute to the development of CS by Streptomyces spp.
The use of antagonist microorganisms against fungal plant pathogens is an attractive and ecologically alternative to the use of chemical pesticides. Streptomyces are beneficial soil bacteria and potential candidates for biocontrol agents. This study reports the isolation, characterization and antagonist activity of soil streptomycetes from the Los Petenes Biosphere Reserve, a Natural protected area in Campeche, Mexico. The results showed morphological, physiological and biochemical characterization of six actinomycetes and their inhibitory activity against Curvularia sp., Aspergillus niger, Helminthosporium sp. and Fusarium sp. One isolate, identified as Streptomyces sp. CACIS-1.16CA showed the potential to inhibit additional pathogens as Alternaria sp., Phytophthora capsici, Colletotrichum sp. and Rhizoctonia sp. with percentages ranging from 47 to 90 %. This study identified a streptomycete strain with a broad antagonist activity that could be used for biocontrol of plant pathogenic fungi.
Rashad, Ferial M; Fathy, Hayam M; El-Zayat, Ayatollah S; Elghonaimy, Ahlam M
Different strategies have been employed for selective isolation of Streptomycetes from 20 marine samples varied in their biological nature. The recovery of Streptomycetes isolates (112) was influenced preferentially by different strategies; sediment samples were the best source of potential candidate Streptomycetes. All isolates exhibited antimicrobial activities with variable spectrum; the most promising isolates (31) were phenotypically characterized and identified as Streptomyces sp.; these isolates exhibited variable capacity for secretion of numerous hydrolytic enzymes such as catalase, protease, amylase, lipase, lecithinase, asparaginase, chitinase and pectinase. All the strains resisted both penicillin and streptomycin, 29 were sensitive to neomycin; the majority of strains (25) showed multiple antibiotic resistance index greater than 0.2; 23, 22 and 13 degraded the shrimp shell, chicken feather and corn cob, respectively, producing bioactive substance(s) which indicates their diversity and their ecological role in the marine ecosystem. At least 28 strains exhibited nematicidal activity in vitro and in vivo against root-knot nematode and supported plant growth. In vitro, the assessed Streptomyces species exhibited the ability to produce gibberellic acid, indole acetic acid, abscisic acid, kinetin and benzyladenine. Except for indole acetic acid, this is the first report concerning the ability of marine Streptomyces to produce such phytohormones and the use of shrimp shell waste as a mono component medium for production of phytohormones. The study is efficacious in selecting effective biodiverse strains of marine Streptomyces that may work under diverse agro-ecological conditions as a useful element in plant nutrition and as biocontrol agents involved in integrated management programs.
Lin, Yanbing; Hao, Xiuli; Johnstone, Laurel; Miller, Susan J.; Baltrus, David A.; Rensing, Christopher; Wei, Gehong
A draft genome sequence of Streptomyces zinciresistens K42, a novel Streptomyces species displaying a high level of resistance to zinc and cadmium, is presented here. The genome contains a large number of genes encoding proteins predicted to be involved in conferring metal resistance. Many of these genes appear to have been acquired through horizontal gene transfer. PMID:22038968
Rapid identification of clinically significant species and taxa of aerobic actinomycetes, including Actinomadura, Gordona, Nocardia, Rhodococcus, Streptomyces, and Tsukamurella isolates, by DNA amplification and restriction endonuclease analysis.
Steingrube, V A; Wilson, R W; Brown, B A; Jost, K C; Blacklock, Z; Gibson, J L; Wallace, R J
A previously described PCR-restriction fragment length polymorphism (RFLP) identification schema for Nocardia that used an amplified 439-bp segment (amplicon) of the 65-kDa heat shock protein gene was evaluated for potential use with isolates of all clinically significant aerobic actinomycetes. The study included 28 reference (American Type Culture Collection) strains and 198 clinical isolates belonging to 20 taxonomic groups. Of these 198 isolates, 188 could be differentiated by this PCR-RFLP method. Amplicons from all aerobic actinomycete isolates lacked BstEII recognition sites, thereby distinguishing them from those of mycobacteria that contain one or more such sites. Of 29 restriction endonucleases, MspI plus HinfI produced RFLP patterns that differentiated 16 of the 20 taxa. A single RFLP pattern was observed for 15 of 20 taxa that included 65% of phenotypically clustered isolates. Multiple patterns were seen with Gordona bronchialis, Nocardia asteroides complex type VI, Nocardia otitidiscaviarum, Nocardia transvalensis, and Streptomyces spp. Streptomyces RFLP patterns were the most heterogeneous (five patterns among 19 isolates), but exhibited a unique HinfI fragment of > 320 bp. RFLP patterns that matched those from type strains of Streptomyces albus, Streptomyces griseus, or Streptomyces somaliensis were obtained from 14 of 19 Streptomyces isolates. Only 10 of 28 isolates of N. otitidiscaviarum failed to yield satisfactory amplicons, while only 6 of 188 (3.2%) clinical isolates exhibited patterns that failed to match one of the 21 defined RFLP patterns. These studies extended the feasibility of using PCR-RFLP analysis as a rapid method for the identification of all clinically significant species and taxa of aerobic actinomycetes.
Loucif, Lotfi; Bendjama, Esma; Gacemi-Kirane, Djamila; Rolain, Jean-Marc
The recent emergence of multidrug-resistant bacteria over the last decade has led to a renewal in the discovery of new antimicrobial drugs. Streptomyces members are practically unlimited sources of new antibiotics. However, the identification of Streptomyces species is difficult and time-consuming. Therefore, there is a need for alternative methods for their rapid identification. In this study, an efficient protocol of identification using Matrix-Assisted Laser Desorption Ionization Time-Of-Flight Mass Spectrometry (MALDI-TOF MS) was developed and applied for the rapid identification of Streptomyces isolates from the El Kala lakes in northeastern Algeria. A collection of 48 Streptomyces isolates were used for this study. The optimized procedure allowed us to obtain specific and reproducible protein spectra for each Streptomyces isolate tested. The spectra generated were used to build a preliminary local database based on their initial 16S rRNA identification. The blind test used for the identification of 20 Streptomyces strains already available in our created database and 20 unknown Streptomyces isolates showed that all (100%) of the Streptomyces strains listed in the database were rapidly (Streptomyces isolates.
Sripreechasak, Paranee; Tamura, Tomohiko; Shibata, Chiyo; Suwanborirux, Khanit; Tanasupawat, Somboon
A novel actinomycete, strain KC-112T, was isolated from soil collected from Similan Islands, Phang-Nga Province, Thailand. The strain exhibited morphological and chemotaxonomic characteristics consistent with those of members of the genus Streptomyces. The formation of smooth spiral spore chains was observed on aerial mycelia. ll-Diaminopimelic acid was detected in whole-cell hydrolysates, but no diagnostic sugars were detected and the strain lacked mycolic acids. The N-acyl type of muramic acid was acetyl. The major menaquinones were MK-9(H8), MK-9(H6) and MK-9(H2). The predominant cellular fatty acids were anteiso-C15 : 0, anteiso-C17 : 0, iso-C16 : 0 and C16 : 0. The polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylinositol, phosphatidylinositol mannoside, an unknown phospholipid, an unknown aminolipid, unknown lipids and an unknown glycolipid. The DNA G+C content was 73 mol%. On the basis of 16S rRNA gene sequence analysis, strain KC-112T was closely related to Streptomyces fumanus NBRC 13042T (98.8 % 16S rRNA gene sequence similarity), Streptomyces anandii NBRC 13438T (98.8 %) and Streptomyces capillispiralis NBRC 14222T (98.8 %). DNA-DNA relatedness values among strain KC-112T and type strains of closely related species were lower than 70 %. On the basis of evidence from this taxonomic study using a polyphasic approach, strain KC-112T represents a novel species of the genus Streptomyces, namely Streptomyces andamanensis sp. nov. The type strain is KC-112T ( = KCTC 29502T = NBRC 110085T = PCU 347T = TISTR 2401T).
Yousif, Ghada; Busarakam, Kanungnid; Kim, Byung-Yong; Goodfellow, Michael
A Streptomyces strain isolated from a mangrove sediment was classified using a polyphasic approach. The organism, isolate GY1(T), was found to have chemical and morphological properties typical of members of the genus Streptomyces. The isolate was shown to form a distinct phyletic line within the Streptomyces radiopugnans 16S rRNA gene subclade and to be closely related to the type strain of Streptomyces fenhuangensis (98.7 % similarity). It is also closely related to the type strain of Streptomyces bakulensis which was also closely related to members of the Streptomyces glaucosporus 16S rRNA gene subclade. Isolate GY1(T) was distinguished readily from the S. barkulensis type strain and from species classified in the S. radiopugnans clade using a combination of morphological and physiological properties, including a requirement for seawater for growth. Based on the genotypic and phenotypic data, it is proposed that isolate GY1(T) (=NCIMB 14980(T), NRRL B-69296(T)) be classified in the genus Streptomyces as Streptomyces mangrovi sp. nov.
Han, Xiufang; Zheng, Jimei; Xin, Di; Xin, Yuhua; Wei, Xuexin; Zhang, Jianli
Two actinobacterial strains, m20(T) and z8, were isolated from soil taken from rainforest areas/tropic forest region, Yunnan Province, south-west China. The 16S rRNA gene sequence similarities and DNA-DNA relatedness values between strains m20(T) and z8 were 100 and 88.2%, respectively, which indicated that these two strains should be classified as the same species. The taxonomic position of the strains was determined by a polyphasic approach. Morphological and chemotaxonomic features of the strains were consistent with those of the genus Streptomyces . A phylogenetic tree based on 16S rRNA gene sequences showed that strains m20(T) and z8 formed an evolutionary branch within the genus Streptomyces and shared relatively high 16S rRNA gene sequence similarity values with other members of this genus, including 'Streptomyces siamensis' NBRC 108799 (98.95%), Streptomyces graminilatus NBRC 108882(T) (98.25%), Streptomyces seoulensis NBRC 16668(T) (98.11%), Streptomyces peucetius ATCC 29050(T) (98.11%) and Streptomyces hygroscopicus subsp. ossamyceticus ATCC 15420(T) (98.11%). DNA-DNA relatedness values between strain m20(T) and the five above-mentioned strains were 56.3, 55.1, 52.8, 50.1 and 48.4%, respectively. On the basis of phenotypic, genotypic and phylogenetic properties, strains m20(T) and z8 could be distinguished from phylogenetically related members of the genus Streptomyces . The isolates thus merit species status within the genus Streptomyces , for which the name http://dx.doi.org/10.1601/nm.6817 Streptomyces albiflavescens sp. nov. is proposed. The type strain is m20(T) ( =CGMCC 4.7111(T) =KCTC 29196(T)). Strain z8 ( =CGMCC 4.7112=KCTC 29197) is a reference strain.
Harunari, Enjuro; Hamada, Moriyuki; Shibata, Chiyo; Tamura, Tomohiko; Komaki, Hisayuki; Imada, Chiaki; Igarashi, Yasuhiro
A novel Gram-stain-positive actinomycete, designated MB-PO13(T), was isolated from a tunicate (Molgula manhattensis) collected in Tokyo Bay, Japan, and its taxonomic position was studied by a polyphasic approach. Phylogenetic analysis based on 16S rRNA gene sequence comparisons revealed that strain MB-PO13(T) was closely related to Streptomyces graminisoli JR-12(T) (99.72% 16S rRNA gene sequence similarity) and Streptomyces shenzhenensis 172115(T) (99.23%). The strain contained LL-diaminopimelic acid in the whole-cell hydrolysate. The predominant menaquinones were MK-9(H8) and MK-9(H6) and the major fatty acids were anteiso-C15:0, iso-C16:0, iso-C14:0 and C16:0. These data supported the affiliation of the novel strain to the genus Streptomyces. Meanwhile, results of DNA-DNA hybridization and physiological and biochemical tests indicated that strain MB-PO13(T) was distinguished from known Streptomyces type strains. Therefore, strain MB-PO13(T) represents a novel species of the genus Streptomyces for which the name Streptomyces hyaluromycini sp. nov. is proposed; the type strain is MB-PO13(T) (=NBRC 110483(T) =DSM 100105(T)).
Classification of Streptomyces griseus (Krainsky 1914) Waksman and Henrici 1948 and related species and the transfer of 'Microstreptospora cinerea' to the genus Streptomyces as Streptomyces yanii sp. nov.
Liu, Zhiheng; Shi, Yanlin; Zhang, Yamei; Zhou, Zhihong; Lu, Zhitang; Li, Wei; Huang, Ying; Rodríguez, Carlos; Goodfellow, Michael
A soil actinomycete, strain 80-133(T), with the non-validly published name 'Microstreptospora cinerea', was the subject of a polyphasic study designed to clarify its taxonomic status. Comparative 16S rRNA gene sequence studies indicated that the organism belonged to the genus Streptomyces, a result in line with previous chemotaxonomic and morphological data. The strain belonged to the Streptomyces griseus clade, but could be distinguished from representatives of species assigned to this taxon by using DNA-DNA relatedness and phenotypic data. In light of these findings, it is proposed that the organism should be recognized as a novel species of the genus Streptomyces. The name proposed for this taxon is Streptomyces yanii sp. nov., with isolate 80-133(T) (=AS 4.1146(T)=JCM 3331(T)) as the type strain. It was also shown that representative strains of Streptomyces argenteolus, Streptomyces caviscabies, S. griseus and Streptomyces setonii belong to the same genomic species and have key phenotypic properties in common. It is proposed that S. caviscabies and S. setonii should be considered as later heterotypic synonyms of S. griseus and that S. argenteolus AS 4.1693(T) should also be assigned to this taxon.
Idris, Hamidah; Labeda, David P; Nouioui, Imen; Castro, Jean Franco; Del Carmen Montero-Calasanz, Maria; Bull, Alan T; Asenjo, Juan A; Goodfellow, Michael
A polyphasic study was undertaken to determine the taxonomic status of a Streptomyces strain which had been isolated from a high altitude Atacama Desert soil and shown to have bioactive properties. The strain, isolate H9(T), was found to have chemotaxonomic, cultural and morphological properties that place it in the genus Streptomyces. 16S rRNA gene sequence analyses showed that the isolate forms a distinct branch at the periphery of a well-delineated subclade in the Streptomyces 16S rRNA gene tree together with the type strains of Streptomyces crystallinus, Streptomyces melanogenes and Streptomyces noboritoensis. Multi-locus sequence analysis (MLSA) based on five house-keeping gene alleles showed that isolate H9(T) is closely related to the latter two type strains and to Streptomyces polyantibioticus NRRL B-24448(T). The isolate was distinguished readily from the type strains of S. melanogenes, S. noboritoensis and S. polyantibioticus using a combination of phenotypic properties. Consequently, the isolate is considered to represent a new species of Streptomyces for which the name Streptomyces aridus sp. nov. is proposed; the type strain is H9(T) (=NCIMB 14965(T)=NRRL B65268(T)). In addition, the MLSA and phenotypic data show that the S. melanogenes and S. noboritoensis type strains belong to a single species, it is proposed that S. melanogenes be recognised as a heterotypic synonym of S. noboritoensis for which an emended description is given.
The identification and classification of species within the genus Streptomyces is difficult because there are presently 576 validly described species and this number increases every year. The value of the application of multilocus sequence analysis scheme to the systematics of Streptomyces species h...
Cheng, Cong; Li, Yu-Qian; Asem, Mipeshwaree Devi; Lu, Chun-Yan; Shi, Xiao-Han; Chu, Xiao; Zhang, Wan-Qin; Di An, Deng-; Li, Wen-Jun
A novel actinobacterial strain, designated LPA192(T), was isolated from a soil sample collected from Lop Nur, Xinjiang Uygur Autonomous Region, Northwest China. A polyphasic approach was used to investigate the taxonomic position of strain LPA192(T). The isolate showed morphological and chemotaxonomic characteristics typical of members of the genus Streptomyces. Peptidoglycan was found to contain LL-diaminopimelic acid as the diagnostic diamino acid. The predominant menaquinones were MK-9(H6) and MK-10(H4). Polar lipids were phosphatidylethanolamine, diphosphatidylglycerol and phosphatidylinositol. Major cellular fatty acids consist of C16:0, anteiso-C15:0 and C18:1 ω9c. The sugar in whole-cell hydrolysates was mannose. Phylogenetic analysis indicated that strain LPA192(T) is closely related to Streptomyces tanashiensis LMG 20274(T) (99.3 %), Streptomyces gulbargensis DAS131(T) (99.3 %), Streptomyces nashvillensis NBRC 13064(T) (99.3 %), Streptomyces roseolus NBRC 12816(T) (99.2 %) and Streptomyces filamentosus NBRC 12767(T) (99.1 %) while showing below 98.5 % sequencing similarities with other validly published Streptomyces species. However, DNA-DNA relatedness values between LPA192(T) and the closely related type strains were below 40 %, which are much lower than 70 % threshold value for species delineation. The genomic DNA G + C content of strain LPA192(T) was 69.3 mol %. Based on the differences in genotypic and phenotypic characteristics from the closely related strains, strain LPA192(T) is considered to represent a novel species of the genus Streptomyces for which the name Streptomyces xinjiangensis sp. nov. is proposed. The type strain is LPA192(T) (=KCTC 39601(T) = CGMCC 4.7288(T)).
Full Text Available Objective(s: Biosynthesis of gold nanoparticles (NGPs is environmentally safer than chemical and physical procedures. This method requires no use of toxic solvents and synthesis of dangerous products and is environmentally safe. In this study, we report the biosynthesis of NGPs using Streptomyces djakartensis isolate B-5. Materials and Methods: NGPs were biosynthesized by reducing aqueous gold chloride solution via a Streptomyces isolate without the need for any additive for protecting nanoparticles from aggregation. We characterized the responsible Streptomycete; its genome DNA was isolated, purified and 16S rRNA was amplified by PCR. The amplified isolate was sequenced; using the BLAST search tool from NCBI, the microorganism was identified to species level. Results: Treating chloroauric acid solutions with this bacterium resulted in reduction of gold ions and formation of stable NGPs. TEM and SEM electro micrographs of NGPs indicated size range from 2- 25 nm with average of 9.09 nm produced intracellular by the bacterium. SEM electro micrographs revealed morphology of spores and mycelia. The amplified PCR fragment of 16S rRNA gene was cloned and sequenced from both sides; it consisted of 741 nucleotides. According to NCBI GenBank, the bacterium had 97.1% homology with Streptomyces djakartensis strain RT-49. The GenBank accession number for partial 16S rRNA gene was recorded as JX162550. Conclusion: Optimized application of such findings may create applications of Streptomycetes for use as bio-factories in eco-friendly production of NGPs to serve in demanding industries and related biomedical areas. Research in this area should also focus on the unlocking the full mechanism of NGPs biosynthesis by Streptomycetes.
Full Text Available Fifteen Streptomyces isolates were isolated from soil in some different location on vegetable plantation at agriculture standard condition. The isolates were assessed for their antibacterial activity against Mycobacterium tuberculosis (MTB ATCC H37RV and mycobacterial which isolated from Dr. Soetomo Hospital patients in Surabaya. The International Streptomyces Project 4 (ISP4 and Middlebrook 7H9 (MB7H9 wwere used as growth or fermentation medium. The screening of inhibition activity was performed using turbidimetry and spot-test on agar medium. Results shown that 33.3% of the isolates (5 isolates have anti-mycobacterial activities. The first line anti tuberculosis drug rifampicin, (RIF, ethambutol (EMB, isoniazid (INH, and pyrazinamide (PZA were used as standards or positive controls with concentration 20 ppm. Optical density of crude fermentation broth concentrated from five isolates relatively lower than five anti-tuberculosis drug activity standard, although their activities against some microbial were similar to the standard at spot-test. The most efficient isolate shown anti-mycobacterial activity was Streptomyces B10 which identified as Streptomyces violaceousniger. In addition, fatty acid methyl ester (FAME profile of gas chromatography-mass spectrometry chromatogram of each isolates were studied and compared to Streptomyces spp. Keywords: Anti-mycobacterial, Mycobacterium tuberculosis, Streptomyces spp.
Sujarit, Kanaporn; Kudo, Takuji; Ohkuma, Moriya; Pathom-Aree, Wasu; Lumyong, Saisamorn
Actinomycete strain CMU-AB204T was isolated from oil palm rhizosphere soil collected in Chiang Mai University (Chiang Mai, Thailand). Based on morphological and chemotaxonomic characteristics, the organism was considered to belong to the genus Streptomyces. Whole cell-wall hydrolysates consisted of ll-diaminopimelic acid, glucose, ribose and galactose. The predominant menaquinones were MK-9(H4), MK-9(H6), MK-9(H2) and MK-8(H4). The fatty acid profile contained iso-C15 : 0, iso-C16 : 0 and anteiso-C15 : 0 as major components. The principal phospholipids detected were phosphatidylethanolamine, diphosphatidylglycerol and phosphatidylglycerol. The DNA G+C content of strain CMU-AB204T was 70.9 mol%. Based on 16S rRNA gene sequence similarity, strain CMU-AB204T was closely related to Streptomyces orinoci JCM 4546T (98.7 %), Streptomyces lilacinus NBRC 12884T (98.5 %), Streptomyces abikoensis CGMCC 4.1662T (98.5 %), Streptomyces griseocarneus JCM 4905T (98.4 %) and Streptomyces xinghaiensis JCM 16958T (98.3 %). Phylogenetic trees revealed that the new strain had a distinct taxonomic position from closely related type strains of the genus Streptomyces. Spiny to hairy spores clearly differentiated strain CMU-AB204T from the five most closely related Streptomyces species, which produced smooth spores. On the basis of evidence from this polyphasic study, it is proposed that strain CMU-AB204T represents a novel species of the genus Streptomyces, namely Streptomyces palmae sp. nov. The type strain is CMU-AB204T (=JCM 31289T=TBRC 1999T).
Zhao, Shanshan; Ye, Lan; Liu, Chongxi; Abagana, Adam Yacoub; Zheng, Weiwei; Sun, Pengyu; Li, Jiansong; Xiang, Wensheng; Wang, Xiangjing
During an investigation exploring potential sources of novel species and natural products, a novel actinomycete with antifungal activity, designated strain NEAU-Gz11(T), was isolated from a soil sample, which was collected from Gama, Chad. The isolate was found to have morphological and chemotaxonomic characteristics typical of members of the genus Streptomyces. 16S rRNA gene sequence similarity studies showed that strain NEAU-Gz11(T) belongs to the genus Streptomyces with high sequence similarity to Streptomyces hiroshimensis JCM 4098(T) (98.0 %). Similarities to other type strains of the genus Streptomyces were lower than 98.0 %. However, the physiological and biochemical characteristics and low levels of DNA-DNA relatedness could differentiate the isolate genotypically and phenotypically from S. hiroshimensis JCM 4098(T). Therefore, the strain is concluded to represent a novel species of the genus Streptomyces, for which the name Streptomyces gamaensis sp. nov. is proposed. The type strain is NEAU-Gz11(T) (=CGMCC 4.7304(T)=DSM 101531(T)).
Pan, Hua-Qi; Cheng, Juan; Zhang, Dao-Feng; Yu, Su-Ya; Khieu, Thi-Nhan; Son, Chu Ky; Jiang, Zhao; Hu, Jiang-Chun; Li, Wen-Jun
A novel actinomycete strain, designated 11A07(T), was isolated from young Scomberomorus niphonius in the Bohai Sea. Basic local alignment search tool analyses showed that this isolate had the highest 16S rRNA gene sequence similarity of 97.41% with Streptomyces rimosus subsp. paromomycinus DSM 41429(T). Phylogenetic tree revealed that strain 11A07(T) formed a distinct lineage clustered with Streptomyces panacagri Gsoil 519(T), Streptomyces sodiiphilus YIM 80305(T) and Streptomyces albus subsp. albus NRRL B-2365(T) having similarities of 97.30%, 97.10% and 96.83%, respectively. Multilocus sequence analysis further demonstrated that the new isolate was different from the selected representatives of Streptomyces as a separate phylogenetic line. Strain 11A07(T) produced straight or rectiflexibile spore chains with smooth surface, white aerial mycelia and brown diffusible pigments on international streptomyces project 2 medium. Maximum tolerated NaCl concentration for growth was 11.0%. Whole-cell sugars were mannose, ribose, glucose, galactose and xylose. The predominant menaquinones were MK-9(H2), MK-9(H4) and MK-9 (H6). The fatty-acid profile contained iso-C16:0, C18:0 10-methyl (tuberculostearic acid) and anteiso-C17:0 as the major compositions. The polar lipids consisted of diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol, phosphatidylinositol mannoside and an unknown phospholipid. The G+C content of the genomic DNA was 71.4 mol%. These morphological, phenotypic and chemotaxonomic properties showed that strain 11A07(T) could be readily distinguished from the most closely related members of the genus Streptomyces. Thus, based on the polyphasic taxonomic data, strain 11A07(T) (=JCM 19630(T)=CCTCC AA 2013020(T)=KCTC 29263(T)) represents a novel species within the genus Streptomyces, for which the name Streptomyces bohaiensis sp. nov. is proposed.
Funk, S.B.; Pasti-Grigsby, M.B.; Felicione, E.C.; Crawford, D.L. [Univ. of Idaho, Moscow, ID (United States)
Composting has been proposed as one process for use in the bioremediation of 2,4,6 trinitrotoluene (TNT)-contaminated soils. However, the biotransformations of TNT that occur during composting, and the specific compost microorganisms involved in TNT metabolism, are not well understood. Both mesophilic and thermophilic actinomycetes are important participants in the biodegradation of organic matter, and possibly TNT, in composts. Here the authors report on the biotransformation of TNT by Streptomyces species growing aerobically in a liquid medium supplemented with 10 to 100 mg/L of TNT. Streptomyces spp. are able to completely remove TNT from the culture medium within 24 hours. As has been observed with other bacteria, these streptomycetes transform TNT first by reducing the 4-nitro and 2-nitro groups to the corresponding amino group; reducing TNT first to 4-amino-2,6-dinitrotoluene and then 2,4-diamino-6-nitrotoluene. These intermediates are transitory and are themselves removed from the medium within 7 days.
Rohland, Jeffrey; Meyers, Paul R
An actinobacterial strain, MV32(T), was isolated from the paunch region of the hindgut of a South African termite, Amitermes hastatus, as part of an investigation of the actinobacterial population residing within this higher order termite species. Strain MV32(T) was chosen for further study from amongst the many potentially novel actinomycete isolates because of its strong antibacterial activity against Mycobacterium aurum A+. 16S rRNA gene phylogenetic analyses clearly placed strain MV32(T) within the genus Streptomyces, with 99.3% sequence similarity to its closest relative, Streptomyces endophyticus YIM 65594(T). Despite this high sequence similarity, DNA-DNA hybridisation analysis showed a DNA relatedness value of 62 ± 2%, to S. endophyticus DSM 41984(T) (indicating that strain MV32(T) belongs to a different genomic species), as well as values of 14.4 ± 0.8 and 10.4 ± 2.9%, respectively, to its next closest relatives, Streptomyces kunmingensis NRRL B-16240(T) and Streptomyces cinnabarinus NRRL B-12382(T). Based on these results and supported by both chemotaxonomic data and a number of phenotypic differences, strain MV32(T) is proposed to represent a new species within the genus Streptomyces, with the name Streptomyces fractus (= DSM 42163(T) = NRRL B-59159(T)).
Silva, Fábio Sérgio Paulino; Souza, Danilo Tosta; Zucchi, Tiago Domingues; Pansa, Camila Cristiane; de Figueiredo Vasconcellos, Rafael Leandro; Crevelin, Eduardo José; de Moraes, Luiz Alberto Beraldo; Melo, Itamar Soares
The taxonomic position of a novel marine actinomycete isolated from a marine sponge, Aplysina fulva, which had been collected in the Archipelago of Saint Peter and Saint Paul (Equatorial Atlantic Ocean), was determined by using a polyphasic approach. The organism showed a combination of morphological and chemotaxonomic characteristics consistent with its classification in the genus Streptomyces and forms a distinct branch within the Streptomyces somaliensis 16S rRNA gene tree subclade. It is closely related to Streptomyces violascens ISP 5183(T) (97.27 % 16S rRNA gene sequence similarity) and Streptomyces hydrogenans NBRC 13475(T) (97.15 % 16S rRNA gene sequence similarity). The 16S rRNA gene similarities between the isolate and the remaining members of the subclade are lower than 96.77 %. The organism can be distinguished readily from other members of the S. violacens subclade using a combination of phenotypic properties. On the basis of these results, it is proposed that isolate 103(T) (=NRRL B-65309(T) = CMAA 1378(T)) merits recognition as the type strain of a new Streptomyces species, namely Streptomyces atlanticus sp. nov.
Phylogenetic analyses of species of Streptomyces based on 16S rRNA gene sequences resulted in a statistically well-supported clade (100% bootstrap value) containing 8 species having very similar gross morphology. These species, including Streptomyces bambergiensis, Streptomyces chlorus, Streptomyces...
Bai, Lu; Liu, Chongxi; Guo, Lifeng; Piao, Chenyu; Li, Zhilei; Li, Jiansong; Jia, Feiyu; Wang, Xiangjing; Xiang, Wensheng
During a screening for novel and biotechnologically useful actinobacteria in insects, a novel actinomycete with antifungal activity, designated strain 1H-GS9(T), was isolated from the head of a Camponotus japonicus Mayr ant, which were collected from Northeast Agricultural University (Harbin, Heilongjiang, China). Strain 1H-GS9(T) was characterised using a polyphasic approach. The organism was found to have morphological and chemotaxonomic characteristics typical of members of the genus Streptomyces. 16S rRNA gene sequence similarity studies showed that strain 1H-GS9(T) belongs to the genus Streptomyces with high sequence similarities to Streptomyces scopuliridis DSM 41917(T) (98.8 %) and Streptomyces mauvecolor JCM 5002(T) (98.6 %). However, phylogenetic analysis based on the 16S rRNA gene sequence indicated that it forms a monophyletic clade with Streptomyces kurssanovii JCM 4388(T) (98.6 %), Streptomyces xantholiticus JCM 4282(T) (98.6 %) and Streptomyces peucetius JCM 9920(T) (98.5 %). Thus, a combination of DNA-DNA hybridization experiments and phenotypic tests were carried out between strain 1H-GS9(T) and the above-mentioned five strains, which further clarified their relatedness and demonstrated that strain 1H-GS9(T) could be distinguished from these strains. Therefore, the strain is concluded to represent a novel species of the genus Streptomyces, for which the name Streptomyces formicae sp. nov. is proposed. The type strain is 1H-GS9(T) (=CGMCC 4.7277(T) = DSM 100524(T)).
Zhang, Binglin; Tang, Shukun; Chen, Ximing; Zhang, Ling; Zhang, Gaoseng; Zhang, Wei; Liu, Guangxiu; Chen, Tuo; Li, Shiweng; Dyson, Paul
A novel actinobacterial strain, designated Z1027T, was isolated from a soil sample collected near the Tuotuo River, Qinghai-Tibet Plateau (China). The strain exhibited antibacterial activity against Escherichia coli and Staphylococcus aureus. The taxonomic position of strain Z1027T was determined using a polyphasic approach. The organism had chemotaxonomic and morphological properties consistent with its classification in the genus Streptomyces and formed a distinct phyletic line in the 16S rRNA gene tree, together with Streptomyces turgidiscabies ATCC 700248T (99.19 % similarity), Streptomyces graminilatus JL-6T (98.84 %) and Streptomyces reticuliscabiei CFBP 4531T (98.36 %). The genomic DNA G+C content of strain Z1027T was 74±1 mol%. The DNA-DNA relatedness values between strain Z1027T and Streptomyces turgidiscabies ATCC 700248T and Streptomyces reticuliscabiei CFBP 4531T were 38.5±0.4 and 26.2±1.2 %, respectively, both of them signiﬁcantly lower than 70 %. Chemotaxonomic data revealed that strain Z1027T possessed MK-9(H6) and MK-9(H8) as the major menaquinones, ll-diaminopimelic acid as the diagnostic diamino acid and galactose as a whole-cell sugar. Diphosphatidylglycerol, phosphatidylethanolamine, phosphatydilinositol and seven other unknown polar lipids were detected; iso-C16 : 0, summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c) and C16 : 0 were the major fatty acids. On the basis of these genotypic and phenotypic data, it is proposed that isolate Z1027T (=CGMCC 4.7272T=JCM 31054T) should be classified as the type strain of a novel species of the genus Streptomyces,Streptomyces lacrimifluminis sp. nov.
Zhou, Shuyu; Li, Zhilei; Bai, Lu; Yan, Kai; Zhao, Junwei; Lu, Chang; Liu, Chongxi; Wang, Xiangjing; Xiang, Wensheng
During an investigation of microbial diversity in medicinal herbs, a novel actinomycete, strain NEAU-QHHV11(T) was isolated from the rhizosphere of Peucedanum praeruptorum Dunn collected from Xianglu Mountain in Heilongjiang Province, northeast China and characterized using a polyphasic approach. The organism was found to have typical characteristics of the genus Streptomyces. Phylogenetic analysis based on 16S rRNA gene sequence also indicated that strain NEAU-QHHV11(T) belongs to the genus Streptomyces and was most closely related to Streptomyces graminilatus NBRC 108882(T) (98.7 % sequence similarity) and Streptomyces turgidiscabies NBRC 16080(T) (98.7 % sequence similarity). The results of DNA-DNA hybridization and some phenotypic characteristics indicated that strain NEAU-QHHV11(T) could be distinguished from its close phylogenetic relatives. Thus, strain NEAU-QHHV11(T) represents a novel species of the genus Streptomyces, for which the name Streptomyces castaneus sp. nov. is proposed. The type strain is NEAU-QHHV11(T) (=CGMCC 4.7235(T) = DSM 100520(T)).
Li, Chuang; Jin, Pinjiao; Liu, Chongxi; Ma, Zhaoxu; Zhao, Junwei; Li, Jiansong; Wang, Xiangjing; Xiang, Wensheng
A novel endophytic actinomycete, designated strain NEAU-HZ10(T) was isolated from moss and characterised using a polyphasic approach. The strain was found to have morphological and chemotaxonomic characteristics typical of the genus Streptomyces. Strain NEAU-HZ10(T) formed grayish aerial mycelia, which differentiated into straight to flexuous chains of cylindrical spores. The cell wall peptidoglycan was found to contain LL-diaminopimelic acid. Predominant menaquinones were identified as MK-9(H6) and MK-9(H8). The polar lipid profile was found to consist of phosphatidylethanolamine, phosphatidylinositol and two unidentified phospholipids. The major fatty acids were identified as iso-C16:0, anteiso-C15:0 and C16:0. 16S rRNA gene sequence similarity studies showed that strain NEAU-HZ10(T) belongs to the genus Streptomyces and exhibits high sequence similarity to Streptomyces cocklensis DSM 42063(T) (98.9 %). Phylogenetic analysis based on the 16S rRNA gene sequence indicated that strain NEAU-HZ10(T) clustered with S. cocklensis DSM 42063(T), Streptomyces yeochonensis CGMCC 4.1882(T) (98.7 %), Streptomyces paucisporeus CGMCC 4.2025(T) (98.4 %) and Streptomyces yanglinensis CGMCC 4.2023(T) (98.1 %). However, a combination of DNA-DNA hybridisation results and some phenotypic characteristics indicated that strain NEAU-HZ10(T) can be distinguished from its phylogenetically closely related strains. Therefore, it is proposed that strain NEAU-HZ10(T) represents a novel species of the genus Streptomyces for which the name Streptomyces bryophytorum sp. nov. is proposed. The type strain is NEAU-HZ10(T) (= CGMCC 4.7151(T) = DSM 42138(T)).
Bouizgarne, B; Lanoot, B; Loqman, S; Spröer, C; Klenk, H-P; Swings, J; Ouhdouch, Y
The novel actinomycete strain Ap1(T) was isolated from rhizosphere soil of the argan tree (Argania spinosa L.) in the south of Morocco. Strain Ap1(T) has been reported as a novel producer of the pentaene polyene macrolide isochainin, which strongly inhibits the growth of pathogenic yeasts and phytopathogenic fungi. Strain Ap1(T) shows a greyish-white aerial mycelium with chains of smooth-surfaced spores of the Spiralis type and a cell wall containing ll-diaminopimelic acid. Based on chemotaxonomy and morphological features, strain Ap1(T) was identified as a member of the genus Streptomyces. 16S rRNA gene sequence similarities based on almost-complete 16S rRNA gene sequences showed that strain Ap1(T) is closely associated with members of the Streptomyces violaceoruber species group (S. violaceoruber, S. coelescens, S. violaceorubidus, 'S. caesius', 'S. lividans', S. violaceolatus and S. humiferus) and others (Streptomyces aurantiogriseus, S. lienomycini, S. chattanoogensis, S. rubrogriseus and S. tendae). However, protein profiling, DNA-DNA hybridization and BOX-PCR fingerprinting proved a relationship above the species level. In addition, the phenotype also allowed for the differentiation of strain Ap1(T) from its closest neighbours. As a result of this polyphasic approach, we conclude that strain Ap1(T) represents a novel species of the genus Streptomyces, for which the name Streptomyces marokkonensis sp. nov. is proposed. The type strain is Ap1(T) (=R-22003(T) =LMG 23016(T) =DSM 41918(T)).
Petrosyan, Pavel; García-Varela, Martin; Luz-Madrigal, Agustín; Huitrón, Carlos; Flores, María Elena
The taxonomic position of a thermophilic actinomycete strain isolated from soil was examined using a polyphasic approach. The strain, designated CH-M-1035T, was assigned to the genus Streptomyces on the basis of chemical and morphological criteria. It formed Rectiflexibiles aerial hyphae that carried long chains of rounded, smooth spores. The almost complete nucleotide sequence of the 16S rRNA gene of strain CH-M-1035T was determined and its comparison with the 16S rDNA sequences of previously studied streptomycetes confirmed the assignment of the novel strain to the genus Streptomyces. Strain CH-M-1035T clustered with species belonging to the Streptomyces thermodiastaticus clade in the 1 6S-rDNA-based phylogenetic tree. However, the phenotypic properties of strain CH-M-1035T differed from those of the recognized species within this clade. Therefore, it is proposed that strain CH-M-1035T be classified as a novel species within the genus Streptomyces, as Streptomyces mexicanus (type strain CH-M-1035T =DSM 41796T =BM-B-384T =NRRL B-24196T).
Tanasupawat, Somboon; Phongsopitanun, Wongsakorn; Suwanborirux, Khanit; Ohkuma, Moriya; Kudo, Takuji
A novel actinomycete, strain RCU-197T, was isolated from soil of a peat swamp forest in Rayong Province, Thailand. Using a polyphasic approach, the strain was classified in the genus Streptomyces. It contained ll-diaminopimelic acid in the cell-wall peptidoglycan. No diagnostic sugars were detected in whole-cell hydrolysates and there was a lack of mycolic acids. The major menaquinones were MK-9(H6) and MK-9(H8). The predominant cellular fatty acids were iso-C14 : 0, iso-C15 : 0, anteiso-C15 : 0 and iso-C16 : 0. The polar lipids profile consisted of diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol, phosphatidylglycerol and phosphatidylinositol mannoside, an unknown aminolipid and two unknown phospholipids. Phylogenetic analysis of 16S rRNA gene sequences showed the strain formed distinct clade within the genus Streptomyces and was closely related to Streptomyces echinatus NBRC 12763T (98.78 % 16S rRNA gene sequence similarity). According to the polyphasic approach as well as DNA-DNA relatedness, the strain could be clearly differentiated from closely related species and represents a novel species of the genus Streptomyces, for which the name Streptomyces actinomycinicus sp. nov. is proposed. The type strain is RCU-197T ( = JCM 30864T = TISTR 2208T = PCU 342T).
Nimaichand, Salam; Zhu, Wen-Yong; Yang, Ling-Ling; Ming, Hong; Nie, Guo-Xing; Tang, Shu-Kun; Ningthoujam, Debananda S; Li, Wen-Jun
A novel actinobacterium, designated MBRL 201(T), was isolated from a sample collected from a limestone quarry at Hundung, Manipur, India. The strain was characterized using polyphasic taxonomy. Comparison of the 16S rRNA gene sequence of strain MBRL 201(T) and other Streptomyces species showed sequence similarities ranging from 93.0 to 99.6 % and strain MBRL 201(T) showed closest similarities to Streptomyces virginiae NBRC 12827(T) (99.6 %) and Streptomyces cinnamonensis NBRC 15873(T) (99.6 %). The DNA relatedness between MBRL 201(T) and the type strains of S. virginiae NBRC 12827(T) and S. cinnamonensis NBRC 15873(T) were 44.5 and 35.6 % respectively. Strain MBRL 201(T) contained LL: -diaminopimelic acid (A(2)pm) as the diagnostic diamino acid, with glucose as the main sugar, while small amounts of galactose, glucose, mannose, rhamnose, ribose and xylose were also present in cell-wall hydrolysates. The major fatty acids identified were anteiso-C(15:0) (38.9 %), iso-C(15:0) (19.9 %) and anteiso-C(17:1) (14.7 %). The predominant menaquinones detected were MK-9(H(6)) and MK-9(H(8)), while the polar lipids were diphosphatidylglycerol, phosphatidylmethylethanolamine, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol and phosphatidylinositolmannosides, with other unknown phospholipids and lipids. The G+C content of the genomic DNA was 72.9 %. The phenotypic and genotypic data showed that strain MBRL 201(T) merits recognition as a representative of a novel species of the genus Streptomyces. It is proposed that the isolate should be classified in the genus Streptomyces as a novel species, Streptomyces manipurensis sp. nov. The type strain is MBRL 201(T) (=DSM 42029(T) = JCM 17351(T)).
Han, Lirong; Zhang, Guoqiang; Miao, Guopeng; Zhang, Xing; Feng, Juntao
A filamentous actinomycete, designated strain ZX01(T), was isolated from forest soil around Kanas Lake of China. A polyphasic taxonomic study was carried out to establish the status of strain ZX01(T). Chemical and morphological properties of the isolate were similar to those of species of the genus Streptomyces. Analysis of the almost complete 16S rRNA gene sequence placed strain ZX01(T) in the genus Streptomyces where it formed a distinct phyletic line with recognized species of this genus. The strain exhibited the highest sequence similarities to Streptomyces lavendofoliae NBRC 12882(T) (99.1%), S. luridus NBRC 12793(T) (99.0%), S. lavendulocolor NBRC 12881(T) (99.0%), S. gobitricini NBRC 15419(T) (99.0%), and S. roseolilacinus NBRC 12815(T) (98.9%). Low DNA-DNA relatedness values of 54.0, 50.0, 60.0, 66.7, and 50.4%, respectively, were found between strain ZX01(T) and corresponding strains above. A number of phenotypic properties also enabled the isolate to be differentiated from related species of the genus Streptomyces. Therefore, it is proposed that strain ZX01(T) should be classified as the type strain of a novel species in the genus Streptomyces, Streptomyces kanasensis sp. nov. The type strain is ZX01(T) (= CGMCC 4893(T) =JCM 30232(T)).
Taxonomic evaluation of Streptomyces albus and related species using multilocus sequence analysis and proposals to emend the description of Streptomyces albus and describe Streptomyces pathocidini sp. nov
In phylogenetic analyses of the genus Streptomyces using 16S rRNA gene sequences, Streptomyces albus subsp. albus NRRL B-1811T forms a cluster with 5 other species having identical or nearly identical 16S rRNA gene sequences. Moreover, the morphological and physiological characteristics of these oth...
Hassan, Ramadan; Shaaban, Mona I; Abdel Bar, Fatma M; El-Mahdy, Areej M; Shokralla, Shadi
Quorum sensing (QS) systems communicate bacterial population and stimulate microbial pathogenesis through signaling molecules. Inhibition of QS signals potentially suppresses microbial infections. Antimicrobial properties of Streptomyces have been extensively studied, however, less is known about quorum sensing inhibitory (QSI) activities of Streptomyces. This study explored the QSI potential of Streptomyces isolated from soil. Sixty-five bacterial isolates were purified from soil samples with morphological characteristics of Streptomyces. The three isolates: S6, S12, and S17, exhibited QSI effect by screening with the reporter, Chromobacterium violaceum. Isolate S17 was identified as Streptomyces coelicoflavus by sequencing of the hypervariable regions (V1-V6) of 16S rRNA and was assigned gene bank number KJ855087. The QSI effect of the cell-free supernatant of isolate S17 was not abolished by proteinase K indicating the non-enzymatic activity of QSI components of S17. Three major compounds were isolated and identified, using spectroscopic techniques (1D, 2D NMR, and Mass spectrometry), as behenic acid (docosanoic acid), borrelidin, and 1H-pyrrole-2-carboxylic acid. 1H-pyrrole-2-carboxylic acid inhibited QS and related virulence factors of Pseudomonas aeruginosa PAO1 including; elastase, protease, and pyocyanin without affecting Pseudomonas viability. At the molecular level, 1H-pyrrole-2-carboxylic acid suppressed the expression of QS genes (lasI, lasR, lasA, lasB, rhlI, rhlR, pqsA, and pqsR). Moreover, QSI activity of S17 was assessed under different growth conditions and ISP2 medium supplemented with glucose 0.4% w/v and adjusted at pH 7, showed the highest QSI action. In conclusion, 1H-pyrrole-2-carboxylic acid, one of the major metabolites of Streptomyces isolate S17, inhibited QS and virulence determinants of P. aeruginosa PAO1. The findings of the study open the scope to exploit the in vivo efficacy of this active molecule as anti-pathogenic and anti
Full Text Available Quorum sensing (QS systems communicate bacterial population and stimulate microbial pathogenesis through signaling molecules. Inhibition of QS signals potentially suppresses microbial infections. Antimicrobial properties of Streptomyces have been extensively studied, however, less is known about quorum sensing inhibitory (QSI activities of Streptomyces. This study explored the QSI potential of Streptomyces isolated from soil. Sixty-five bacterial isolates were purified from soil samples with morphological characteristics of Streptomyces. The three isolates: S6, S12, and S17, exhibited QSI effect by screening with the reporter, Chromobacterium violaceum. Isolate S17 was identified as Streptomyces coelicoflavus by sequencing of the hypervariable regions (V1-V6 of 16S rRNA and was assigned gene bank number KJ855087. The QSI effect of the cell-free supernatant of isolate S17 was not abolished by proteinase K indicating the non-enzymatic activity of QSI components of S17. Three major compounds were isolated and identified, using spectroscopic techniques (1D, 2D NMR and Mass spectrometry, as behenic acid (docosanoic acid, borrelidin and 1H-pyrrole-2-carboxylic acid. 1H-pyrrole-2-carboxylic acid inhibited QS and related virulence factors of Pseudomonas aeruginosa PAO1 including; elastase, protease and pyocyanin without affecting Pseudomonas viability. At the molecular level, 1H-pyrrole-2-carboxylic acid suppressed the expression of QS genes (lasI, lasR, lasA, lasB, rhlI, rhlR, pqsA and pqsR. Moreover, QSI activity of S17 was assessed under different growth conditions and ISP2 medium supplemented with glucose 0.4% w/v and adjusted at pH 7, showed the highest QSI action. In conclusion, 1H-pyrrole-2-carboxylic acid, one of the major metabolites of Streptomyces isolate S17, inhibited QS and virulence determinants of P. aeruginosa PAO1. The findings of the study open the scope to exploit the in vivo efficacy of this active molecule as anti-pathogenic and
Santos, Suikinai Nobre; Gacesa, Ranko; Taketani, Rodrigo Gouvêa; Long, Paul F; Melo, Itamar Soares
The genome sequence of the first Streptomyces species isolated from the Brazilian Caatinga is reported here. Genes related to environmental stress tolerance were prevalent and included many secondary metabolic gene clusters. Copyright © 2015 Santos et al.
Lee, Hyo-Jin; Whang, Kyung-Sook
A novel strain of Gram-staining-positive actinobacterium, designated strain JR-39T, was isolated from the rhizosphere soil of bamboo (Sasa borealis) sampled in Damyang, Korea, and its taxonomic position was investigated by a polyphasic approach. The isolate formed flexuous chains of spores that were cylindrical and smooth-surfaced. Strain JR-39T grew at 4–37 °C (optimum 28 °C). The pH range for growth was pH 5–10 (optimum pH 6–8) and the NaCl range for growth was 0–5 % (w/v) with optimum growth at 1 % NaCl. The cell-wall peptidoglycan contained ll-diaminopimelic acid, glutamic acid, alanine and glycine. Whole-cell hydrolysates mainly contained glucose, mannose, ribose and rhamnose. Predominant menaquinones were MK-9 (H6), MK-9 (H8) and MK-9 (H4). The major cellular fatty acids were anteiso-C15 : 0, iso-C16 : 0, iso-C15 : 0 and iso-C14 : 0. The G+C content of the DNA was 72.3 ± 0.34 mol%. Phylogenetic analyses based on 16S rRNA gene sequence analysis indicated that strain JR-39T belonged to the genus Streptomyces, showing the highest sequence similarity to Streptomyces panaciradicis 1MR-8T (99.4 %), Streptomyces capoamus JCM 4734T (98.8 %), Streptomyces galbus DSM 40089T (98.7 %), Streptomyces longwoodensis LMG 20096T (98.7 %), Streptomyces bungoensis NBRC 15711T (98.7 %) and Streptomyces rhizophilus JR-41T (98.7 %). However, DNA–DNA hybridization assays, as well as physiological and biochemical analyses, showed that strain JR-39T could be differentiated from its closest phylogenetic relatives. On the basis of the phenotypic and genotypic characteristics, strain JR-39T represents a novel species for which the name Streptomyces sasae sp. nov. is proposed. The type strain is JR-39T ( = KACC 17182T = NBRC 109809T).
Guan, Xuejiao; Li, Wenchao; Liu, Chongxi; Jin, Pinjiao; Guo, Siyu; Wang, Xiangjing; Xiang, Wensheng
During a screening for novel and biotechnologically useful actinobacteria, a novel actinobacteria with weak antifungal activity, designated strain NEAU-Spg19(T), was isolated from a soil sample collected from pine forest in Songpinggou, Sichuan, southwest China. The strain was characterized using a polyphasic taxonomic approach which confirmed that it belongs to the genus Streptomyces. Growth occurred at a temperature range of 10-30 °C, pH 5.0-11.0 and NaCl concentrations of 0-5 %. The cell wall peptidoglycan consisted of LL-diaminopimelic acid and glycine. The major menaquinones were MK-9(H6), MK-9(H8) and MK-9(H4). The phospholipid profile contained diphosphatidylglycerol (DPG), phosphatidylethanolamine and phosphatidylinositol. The major fatty acids were iso-C15:0, iso-C16:0, and C16:0. 16S rRNA gene sequence similarity studies showed that strain NEAU-Spg19(T) belongs to the genus Streptomyces with the highest sequence similarities to Streptomyces tauricus JCM 4837(T) (98.6 %) and Streptomyces rectiviolaceus JCM 9092(T) (98.3 %). Some physiological and biochemical properties and low DNA-DNA relatedness values enabled the strain to be differentiated from S. tauricus JCM 4837(T) and S. rectiviolaceus JCM 9092(T). Hence, on the basis of phenotypic and genetic analyses, it is proposed that strain NEAU-Spg19(T) represents a novel species of the genus Streptomyces, for which the name Streptomyces songpinggouensis sp. nov. is proposed. The type strain is NEAU-Spg19(T) (=CGMCC 4.7140(T)=DSM 42141(T)).
Amin, Arshia; Ahmed, Iftikhar; Khalid, Nauman; Osman, Ghenijan; Khan, Inam Ullah; Xiao, Min; Li, Wen-Jun
A Gram-staining positive, non-motile, rod-shaped, catalase positive and oxidase negative bacterium, designated NCCP-1331(T), was isolated from a hot water spring soil collected from Tatta Pani, Kotli, Azad Jammu and Kashmir, Pakistan. The isolate grew at a temperature range of 18-40 °C (optimum 30 °C), pH 6.0-9.0 (optimum 7.0) and with 0-6 % NaCl (optimum 2 % NaCl (w/v)). The phylogenetic analysis based on 16S rRNA gene sequence revealed that strain NCCP-1331(T) belonged to the genus Streptomyces and is closely related to Streptomyces brevispora BK160(T) with 97.9 % nucleotide similarity, followed by Streptomyces drosdowiczii NRRL B-24297(T) with 97.8 % nucleotide similarity. The DNA-DNA relatedness values of strain NCCP-1331(T) with S. brevispora KACC 21093(T) and S. drosdowiczii CBMAI 0498(T) were 42.7 and 34.7 %, respectively. LL-DAP was detected as diagnostic amino acid along with alanine, glycine, leucine and glutamic acid. The isolate contained MK-9(H8) as the predominant menaquinone. Major polar lipids detected in NCCP-1331(T) were phosphatidylethanolamine, phosphatidylinositol and unidentified phospholipids. Major fatty acids were iso-C16: 0, summed feature 8 (18:1 ω7c/18:1 ω6c), anteiso-C15:0 and C16:0. The genomic DNA G + C content was 69.8 mol %. On the basis of phylogenetic, phenotypic and chemotaxonomic analysis, it is concluded that strain NCCP-1331(T) represents a novel species of the genus Streptomyces, for which the name Streptomyces caldifontis sp. nov. is proposed. The type strain is NCCP-1331(T) (=KCTC 39537(T) = CPCC 204147(T)).
Hamm, Paris S.; Caimi, Nicole A.; Northup, Diana E.; Valdez, Ernest W.; Buecher, Debbie C.; Dunlap, Christopher A.; Labeda, David P.; Lueschow, Shiloh; Porras-Alfaro, Andrea
At least two-thirds of commercial antibiotics today are derived from Actinobacteria, more specifically from the genus Streptomyces. Antibiotic resistance and new emerging diseases pose great challenges in the field of microbiology. Cave systems, in which actinobacteria are ubiquitous and abundant, represent new opportunities for the discovery of novel bacterial species and the study of their interactions with emergent pathogens. White-nose syndrome is an invasive bat disease caused by the fungus Pseudogymnoascus destructans, which has killed more than six million bats in the last 7 years. In this study, we isolated naturally occurring actinobacteria from white-nose syndrome (WNS)-free bats from five cave systems and surface locations in the vicinity in New Mexico and Arizona, USA. We sequenced the 16S rRNA region and tested 632 isolates from 12 different bat species using a bilayer plate method to evaluate antifungal activity. Thirty-six actinobacteria inhibited or stopped the growth of P. destructans, with 32 (88.9%) actinobacteria belonging to the genus Streptomyces. Isolates in the genera Rhodococcus, Streptosporangium, Luteipulveratus, and Nocardiopsis also showed inhibition. Twenty-five of the isolates with antifungal activity against P. destructans represent 15 novel Streptomyces spp. based on multilocus sequence analysis. Our results suggest that bats in western North America caves possess novel bacterial microbiota with the potential to inhibit P. destructans.
Nguyen, uan Manh; Kim, Jaisoo
This study describes a novel actinomycete, designated T113T, which was isolated from forest soil in Pyeongchang-gun, Republic of Korea, and is an aerobic, Gram-stain-positive actinobacterium that forms flexibilis chains of smooth, elliptical or short rod-shaped spores. The results of 16S rRNA sequence analysis indicated that strain T113T exhibited high levels of similarity to previously characterized species of the genus Streptomyces (98.19–98.89 %, respectively). However, the results of phylogenetic and DNA–DNA hybridization analyses confirmed that the organism represented a novel member of the genus Streptomyces. Furthermore, using chemotaxonomic and phenotypic analyses it was demonstrated that the strain exhibited characteristics similar to those of other members of the genus Streptomyces. The primary cellular fatty acids expressed by this strain included anteiso-C15 : 0, anteiso-C17 : 0, iso-C15 : 0 and iso-C16 : 0. While diphosphatidylglycerol and phosphatidylethanolamine were the predominant lipids expressed by strain T113T, moderate amounts of phosphatidylinositol and phosphatidylinositol mannoside were also detected. Whole-cell hydrolysates contained glucose and ribose, and the predominant menaquinone detected was MK-9 (H6); however, moderate amounts of MK-9 (H8) and trace amounts of MK-10 (H2) and MK-10 (H4) were also detected. We therefore propose that strain T113T be considered as representing a novel species of the genus Streptomyces and propose the name Streptomyces gilvifuscus sp. nov. for this species, with strain T113T ( = KEMB 9005-213T = KACC 18248T = NBRC 110904T) being the type strain.
Dec 18, 2013 ... series on all media. The results showed that glycerol yeast extract and starch casein were the best .... MATERIALS AND METHODS. Collection of .... Genomic DNA was isolated from the isolates using a bacterial genomic DNA ...
Meysam Soltani Nejad
Full Text Available Objective(s: In recent years, the biosynthesis of gold nanoparticles has been the focus of interest because of their emerging application in a number of areas such as biomedicine. In the present study we report the extracellular biosynthesis of gold nanoparticles (AuNPs by using a positive bacterium named Streptomyces fulvissimus isolate U from rice fields of Guilan Province, Iran. Materials and Methods: From over 20 Streptomyces isolates collected, isolate U showed high AuNPs biosynthesis activity. To determine its taxonomical identity, its morphology was characterized by scanning electron microscope and partial molecular analysis performed by PCR. In this regard, 16S rDNA of isolate U was amplified using universal bacterial primers FD1 and RP2. The PCR products were purified and sequenced. Sequence analysis of 16S rDNA was then conducted using NCBI BLAST method. In biosynthesis of AuNPs by this bacterium, the biomass of bacterium exposed to the HAuCl4 solution. Results: The nanoparticles obtained were characterized by UV-Visible spectroscopy, transmission electron microscopy (TEM and Energy dispersive X-ray (EDX spectroscopy and X-ray diffraction spectroscopy (XRD analyses. Our results indicated that Streptomyces fulvissimus isolateU bio-synthesizes extracellular AuNPs in the range of 20-50 nm. Conclusions: This technique of green synthesis of AuNPs by a microbial source may become a promising method because of its environmental safety. Its optimization may make it a potential procedure for industrial production of gold nanoparticles.
TREJO, W H; BENNETT, R E
Trejo, W. H. (Squibb Institute for Medical Research, New Brunswick, N.J.) and R. E. Bennett. Streptomyces species comprising the blue-spore series. J. Bacteriol. 85:676-690. 1963.-The objective of this study was to define and delimit the streptomycetes of the blue-spored (Viridochromogenes) series. The series, as defined in this study, includes 11 blue and blue-green species. The green-spored species were excluded on the basis of morphology as well as color. It was proposed that NRRL B-1511 be designated as the neotype strain of Streptomyces viridochromogenes (Krainsky) Waksman and Henrici, and that IMRU 3761 be designated as the neotype for Streptomyces cyaneus (Krassilnikov) Waksman. Evidence was presented to show that physiological criteria cannot be used to differentiate these organisms below the series level. The major characteristics of the Viridochromogenes series are blue to blue-green spores borne in spirals, and chromogenicity (melanin-positive). Reverse color and spore morphology provide a basis for separation below the series level.
Araujo, Ricardo; Adetutu, Eric; Tobe, Shanan S.; Mallya, Sandeep; Paul, Bobby; Satyamoorthy, Kapaettu
The complete genome sequences of three endophytic Streptomyces species were compared. Strains EN16, EN23, and EN27 were isolated from surface-sterilized roots of wheat plants from South Australia. In field trials, these strains are effective in suppressing fungal root diseases of wheat when added as spore coatings to wheat seed. PMID:27932645
Guan, Xuejiao; Liu, Chongxi; Zhao, Junwei; Fang, Baozhu; Zhang, Yuejing; Li, Lianjie; Jin, Pinjiao; Wang, Xiangjing; Xiang, Wensheng
A novel actinomycete, designated strain NEAU-Spg16(T), was isolated from a soil sample from a pine forest in Songpinggou, Maoxian, southwest China. A polyphasic taxonomic study was carried out to establish the status of strain NEAU-Spg16(T). The cell wall peptidoglycan was found to contain LL-diaminopimelic acid and glycine. The major menaquinones were identified as MK-9(H8), MK-9(H4) and MK-9(H6). The phospholipid profile was found to contain diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol and an unidentified phospholipid. The major fatty acids were identified as iso-C(16:0), C(18:0), C(16:0) and iso-C(15:0). 16S rRNA gene sequence similarity studies showed that strain NEAU-Spg16(T) belongs to the genus Streptomyces with the highest sequence similarity to Streptomyces nitrosporeus DSM 40023(T) (98.6%). However, phylogenetic analysis based on the 16S rRNA gene sequence indicated that it is most closely related to Streptomyces scopuliridis DSM 41917(T) (98.2% sequence similarity). A combination of DNA-DNA hybridization results and some phenotypic characteristics indicated that strain NEAU-Spg16(T) can be clearly differentiated from S. scopuliridis DSM 41917(T) and S. nitrosporeus DSM 40023(T). Therefore, it is concluded that strain NEAU-Spg16(T) represents a novel species of the genus of Streptomyces, for which the name Streptomyces maoxianensis sp. nov. is proposed. The type stain is NEAU-Spg16(T) (=CGMCC 4.7139(T) = DSM 42137(T)).
Zainal, Nurullhudda; Ser, Hooi-Leng; Yin, Wai-Fong; Tee, Kok-Keng; Lee, Learn-Han; Chan, Kok-Gan
A novel Streptomyces strain, MUSC 119(T), was isolated from a soil collected from a mangrove forest. Cells of MUSC 119(T) stained Gram-positive and formed light brownish grey aerial mycelium and grayish yellowish brown substrate mycelium on ISP 2 medium. A polyphasic approach was used to determine the taxonomic status of strain MUSC 119(T), which shows a range of phylogenetic and chemotaxonomic properties consistent with those of the genus Streptomyces. The cell wall peptidoglycan consisted of LL-diaminopimelic acid. The predominant menaquinones were identified as MK-9(H8), MK-9(H6) and MK-9(H4). The polar lipid profile consisted of phosphatidylinositol, phosphatidylethanolamine, glycolipids, diphosphatidylglycerol and four phospholipids. The predominant cellular fatty acids were anteiso-C15:0, iso-C16:0, and anteiso-C17:0. The cell wall sugars were glucose, mannose, ribose and rhamnose. The phylogenetic analysis based on 16S rRNA gene sequence similarity showed that strain MUSC119(T) to be closely related to Streptomyces rhizophilus JR-41(T) (99.0 % sequence similarity), S. panaciradicis 1MR-8(T) (98.9 %), S. gramineus JR-43(T) (98.8 %) and S. graminisoli JR-19(T) (98.7 %). These results suggest that MUSC 119(T) should be placed within the genus Streptomyces. DNA-DNA relatedness values between MUSC 119(T) to closely related strains ranged from 14.5 ± 1.3 to 27.5 ± 0.7 %. The G+C content was determined to be 72.6 mol %. The polyphasic study of MUSC 119(T) showed that this strain represents a novel species, for which the name Streptomyces humi sp. nov. is proposed. The type strain of S. humi is MUSC 119(T) (=DSM 42174(T) = MCCC 1K00505(T)).
Full Text Available Pendem village in Jembrana regency is one of the banana plantation in Bali. Now a days banana plants were attack by bacterial wilt disease with the symptoms of wilting plants, brown spots on the vessel banana stems and fruit to rot and dry. Control of use of chemical fertilizers can cause bad impact on environment and also can not control the disease. Streptomyces bacteria are bacteria that are capable of producing enzymes and antibiotics that can be used as biocontrol agents of several diseases in plants. The purpose of this research is to isolate and identify the bacteria Streptomyces from rhizosphere of banana plants without symptoms in the village Pendem Jembrana regency. The method of isolation of Streptomyces using Platting method, Streptomyces isolated from soil rhizosphere of banana plants without symptoms or health plant. Soil was taken by digging near rooting bananas plant about 15 cm from the ground and and the sample was growth on media Humic Vitamin Agar (HVA and Yeast Extract Malt Agar (ISP4. Identification macros-copically and microscopically and biochemical test using determination key book guide to the Classification and Identification of the Actinomycetes and Their antibiotics of Lechevalier and Waksman (1973. Result showed it was found 9 Streptomyces isolate; Streptomyces sp.1, Streptomyces sp. 2, Streptomyces sp.3, sp.4 Streptomyces, Streptomyces sp.5 sp.6, Streptomyces sp 7, Streptomyces sp.8 and Streptomyces sp.9. Nine isolates of Streptomyces sp. will be tested against the bacteria Ralstonia solanacearum ,the bacteria that causes bacterial wilt disease.
le Roes-Hill, Marilize; Meyers, Paul R
As part of an antibiotic-screening programme, an actinomycete, designated strain SPR(T), was isolated from soil collected from the banks of the Umgeni River, KwaZulu-Natal Province, South Africa. The isolate produced branching vegetative mycelia with sporangiophores bearing sporangia developing at a late stage of growth. The sporangia contained smooth, almond-shaped, non-motile spores. Strain SPR(T) exhibited antibiosis against various Gram-positive and Gram-negative bacteria, including Enterococcus faecium VanA (a vancomycin-resistant strain), Mycobacterium aurum A+ and Escherichia coli ATCC 25922. The chemotaxonomic characteristics of the strain, with the exception of the phospholipid pattern, corresponded with those of the members of the family Streptomycetaceae Waksman and Henrici 1943. Furthermore, phylogenetic analysis based on 16S rRNA genes showed that the strain was closely related to members of the genus Streptomyces, which supports its classification in the family Streptomycetaceae. Thus strain SPR(T) represents a novel species of the genus Streptomyces, for which the name Streptomyces polyantibioticus sp. nov. is proposed. The type strain is SPR(T) (=DSM 44925(T)=NRRL B-24448(T)).
Huang, Mei-Juan; Fei, Jing-Jing; Salam, Nimaichand; Kim, Chang-Jin; Hozzein, Wael N; Xiao, Min; Huang, Hai-Quan; Li, Wen-Jun
An actinomycete strain, designated YIM T102(T), was isolated from the rhizospheric soil of Psammosilene tunicoides W. C. Wu et C. Y. Wu collected from Lijiang, Yunnan Province, China. The taxonomic position of the new isolate was investigated by a polyphasic approach. Phylogenetic analyses based on 16S rRNA gene sequences indicated that strain YIM T102(T) belongs to the genus Streptomyces. Strain YIM T102(T) was most closely related to Streptomyces eurocidicus NRRL B-1676(T) with a pairwise 16S rRNA gene sequence similarity of 98.9 %. However, DNA-DNA relatedness value between strain YIM T102(T) and S. eurocidicus NBRC 13491(T) was found to be 37.8 ± 1.8 %. The menaquinone composition detected for strain YIM T102(T) was MK-9 (H6) and MK-9 (H8), while the major fatty acids were summed feature 4 (38.0 %), anteiso-C15:0 (13.1 %), iso-C16:0 (10.1 %), summed feature 3 (9.8 %) and C16:0 (9.0 %) and iso-C15:0 (5.2 %). The whole-cell hydrolysates contained galactose, glucose, ribose and mannose, along with LL-diaminopimelic acid as the diagnostic diamino acid in the peptidoglycan. The DNA G+C content was 70.7 mol%. Strain YIM T102(T) also exhibited antagonistic activity against Alternaria alternata, Alternaria brassicae and Colletotrichum nicotianae Averna, based on the findings from the comparative analyses of phenotypic and genotypic characteristics; it is proposed that strain YIM T102 represents a novel species of the genus Streptomyces, for which the name Streptomyces zhihengii sp. nov. is proposed. The type strain is YIM T102(T) (=KCTC 39115(T) = DSM 42176(T) = CGMCC 4.7248(T)).
Mo, Ping; Yu, Yi-Zun; Zhao, Jia-Rong; Gao, Jian
An actinomycete strain, designated strain LUSFXJ(T), was isolated from a soil sample obtained near the Xiangtan Manganese Mine, Central-South China and characterised using a polyphasic taxonomic approach. The 16S rRNA gene sequence-based phylogenetic analysis indicated that this strain belongs to the genus Streptomyces. The DNA-DNA relatedness between this strain and two closely related type strains, Streptomyces echinatus CGMCC 4.1642(T) and Streptomyces lanatus CGMCC 4.137(T), were 28.7 ± 0.4 and 19.9 ± 2.0%, respectively, values which are far lower than the 70% threshold for the delineation of a novel prokaryotic species. The DNA G+C content of strain LUSFXJ (T) is 75.0 mol%. Chemotaxonomic analysis revealed that the menaquinones of strain LUSFXJ(T) are MK-9(H6), MK-9(H8), MK-9(H2) and MK-8(H8). The polar lipid profile of strain LUSFXJ(T) was found to contain diphosphatidylglycerol and an unidentified polar lipid. The major cellular fatty acids were identified as iso-C15:0, anteiso-C15:0, iso-C16:0, C16:0 and Summed feature 3. Strain LUSFXJ(T) was found to contain meso-diaminopimelic acid as the diagnostic cell wall diamino acid and the whole cell hydrolysates were found to be rich in ribose, mannose and glucose. Based on phenotypic, phylogenetic and chemotaxonomic characteristics, it is concluded that strain LUSFXJ(T) represents a novel species of the genus Streptomyces, for which the name S. xiangtanensis sp. nov. is proposed. The type strain is LUSFXJ(T) (=GDMCC 4.133(T) = KCTC 39829(T)).
Luo, Xiaoxia; Sun, Yong; Xie, Sinan; Wan, Chuanxing; Zhang, Lili
A novel actinobacterium, designated TRM 43006T, was isolated from the rhizosphere soil of Populus euphratica in Xinjiang Province, north-west China. Phylogenetic and phenotypic analysis demonstrated that strain TRM 43006T belongs to the genus Streptomyces. The strain was aerobic and Gram-stain-positive; the aerial mycelium branched monopodially, forming chains of arthrospores. The spores were oval to cylindrical with smooth surfaces. The whole-cell sugar pattern of strain TRM 43006T consisted of xylose, mannitol, galactose and ribose. The menaquinones were MK-9(H6), MK-9(H8) and MK-9(H10). The polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine, phosphatidylinositol, phosphatidylinositol mannosides and four unknown phospholipids. Major fatty acids were iso-C16 : 0, iso-C16 : 1, iso-C14 : 0 and anteiso-C15 : 0. The G+C content of the genomic DNA was 69.0 mol%. Comparative 16S rRNA gene sequence analysis indicated that strain TRM 43006T was phylogenetically most closely related to Streptomyces roseolilacinus NBRC 12815T (98.6 % similarity) and Streptomycessudanensis SD 504T (98.3 %); however, DNA-DNA hybridization studies between S. roseolilacinus NBRC 12815T, S. sudanensis SD 504T and TRM 43006T showed only 30.28 and 30.65 % relatedness, respectively. Based on the evidence from this polyphasic study, strain TRM 43006T represents a novel species of the genus Streptomyces, for which the name Streptomyces indoligenes sp. nov. is proposed. The type strain is TRM 43006T (=KCTC 39611T=CCTCC AA 2015010T).
Akhwale, Juliah Khayeli; Göker, Markus; Rohde, Manfred; Spröer, Cathrin; Schumann, Peter; Klenk, Hans-Peter; Boga, Hamadi Iddi
A novel strain, designated No. 7(T), was isolated from a sediment sample collected from the alkaline, saline Lake Elmenteita located in the Kenyan Rift Valley. The optimal growth for the strain was found to be at temperature 30-35 °C, at pH 8.0-12.0 in the presence of 7.0-10.0 % (w/v) NaCl. The strain was observed to form a light green beige abundant aerial mycelium on Horikoshi 1 agar and to have morphological and chemotaxonomic characteristics typical of members of the genus Streptomyces. The peptidoglycan was found to contain LL-diaminopimelic acid as the diamino acid, with no diagnostic sugars identified. The predominant menaquinone was identified as MK-9(H6). The main polar lipids were identified as diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol and an unknown phospholipid. Cellular fatty acids were found to consist of saturated branched-chain acids with iso-C(15:0), anteiso-C(15:0), iso-C(16:0) and anteiso-C(17:0) acids predominating. The type strain had a genomic DNA G+C content of 72.8 mol% and formed a distinct phyletic line within the genus Streptomyces. Based on the chemotaxonomic results, 16S rRNA gene sequence analysis and the low DNA-DNA hybridization value with the type strain of Streptomyces calidiresistens, it is proposed that strain No. 7(T) (= DSM 42118 = CECT 8549) represents a novel species, Streptomyces alkaliphilus. The INSDC accession number for the 16S rRNA gene sequence of strain No. 7(T) is KF976730.
Sharma, Trupti K; Mawlankar, Rahul; Sonalkar, Vidya V; Shinde, Vidhya K; Zhan, Jing; Li, Wen-Jun; Rele, Meenakshi V; Dastager, Syed G; Kumar, Lalitha Sunil
A novel alkaliphilic actinomycete, strain NCL716(T), was isolated from a soil sample collected from the vicinity of Lonar Lake, an alkaline salt water meteorite lake in Buldhana district of Maharashtra State in India. The strain was characterised using a polyphasic taxonomic approach which confirmed that it belongs to the genus Streptomyces. Growth was observed over a pH range of 7-11 at 28 °C. The cell wall was found to contain LL-diaminopimelic acid and traces of meso-diaminopimelic acid. The major fatty acid components were identified as iso-C16:0 (46.8 %), C17:1 (12.4 %), anteiso-C15:0 (5.1 %) and anteiso-C17:1 (4.8 %). The major polar lipids were identified as diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol and phosphatidylinositol. The major menaquinones were determined to be MK-9 (H6) (70.3 %), MK-9 (H4) (15.5 %) and MK-9 (H8) (7.2 %). The G+C content of the DNA of the type strain was determined to be 71.4 mol %. The 16S rRNA gene sequence has been deposited in GenBank with accession number FJ919811. Although the 16S rRNA gene sequence analysis revealed that strain NCL716(T) shares >99 % similarity with that of Streptomyces bohaiensis strain 11A07(T), DNA-DNA hybridization revealed only 33.2 ± 3.0 % relatedness between them. Moreover, these two strains can be readily distinguished by some distinct phenotypic characteristics. Hence, on the basis of phenotypic and genetic analyses, it is proposed that strain NCL716(T) represents a novel species of the genus Streptomyces, for which the name Streptomyces lonarensis sp. nov., is proposed. The type strain is NCL 716(T) (=DSM 42084(T) = MTCC 11708(T) = KCTC 39684(T)).
Sultanpuram, Vishnuvardhan Reddy; Mothe, Thirumala; Mohammed, Farooq
An alkaliphilic actinomycete, strain AC3(T), was isolated from Lonar soda lake, in India. Based on 16S rRNA gene sequence analysis it was identified that the strain belongs to the class Actinobacteria and was most closely related to Streptomyces sodiiphilus JCM 13581(T) (96.4 % sequence similarity), Streptomyces leeuwenhoekii DSM 42122(T) (96.1 %), Streptomyces albus NRRL B-2365(T) (96.1 %), Streptomyces panacagri Gsoil 519(T) (96.0 %), Streptomyces fimbriatus NBRC 15411(T) (95.9 %) and other members of the genus Streptomyces (cream substrate and white aerial mycelia on most tested media. The optimum pH for growth was determined to be 9.5-10.0 with no growth at pH 7.0. The DNA G+C content of strain AC3(T) was determined to be 71.2 mol %. The results of the polyphasic analysis allowed a clear differentiation of strain AC3(T) from all other members of the genus Streptomyces. Strain AC3(T) is thus considered to represent a novel member of the genus Streptomyces, for which the name Streptomyces alkalithermotolerans sp. nov. is proposed. The type strain is AC3(T) (=KCTC 29497(T) = JCM 30167(T)).
In phylogenetic analyses of the genus Streptomyces using 16S rRNA gene sequences, Streptomyces albus subsp. albus NRRL B-1811T formed a cluster with 5 other species having identical or nearly identical 16S rRNA gene sequences. Moreover, the morphological and physiological characteristics of these ot...
Baskaran, R; Mohan, P M; Sivakumar, K; Kumar, Ashok
Actinomycetes, especially species of Streptomyces are prolific producers of pharmacologically significant compounds accounting for about 70% of the naturally derived antibiotics that are presently in clinical use. In this study, we used five solvents to extract the secondary metabolites from marine Streptomyces parvulus DOSMB-D105, which was isolated from the mangrove sediments of the South Andaman Islands. Among them, ethyl acetate crude extract showed maximum activity against 11 pathogenic bacteria and six fungi. Presence of bioactive compounds in the ethyl acetate extract was determined using GC-MS and the compounds detected in the ethyl acetate extract were matched with the National Institute of Standards and Technology (NIST) library. Totally eight compounds were identified and the prevalent compounds were 2 steroids, 2 alkaloids, 2 plasticizers, 1 phenolic and 1 alkane. Present study revealed that S. parvulus DOSMB-D105 is a promising species for the isolation of valuable bioactive compounds to combat pathogenic microbes.
Labeda, David P; Rong, Xiaoying; Huang, Ying; Doroghazi, James R; Ju, Kou-San; Metcalf, William W
Previous phylogenetic analysis of species of the genus Streptomyces based on 16S rRNA gene sequences resulted in a statistically well-supported clade (100 % bootstrap value) containing eight species that exhibited very similar gross morphology in producing open looped (Retinaculum-Apertum) to spiral (Spira) chains of spiny- to hairysurfaced, dark green spores on their aerial mycelium. The type strains of the species in this clade, specifically Streptomyces bambergiensis, Streptomyces cyanoalbus, Streptomyces emeiensis, Streptomyces hirsutus, Streptomyces prasinopilosus and Streptomyces prasinus, were subjected to multi-locus sequence analysis (MLSA) utilizing partial sequences of the housekeeping genes atpD, gyrB, recA, rpoB and trpB to clarify their taxonomic status. The type strains of several recently described species with similar gross morphology, including Streptomyces chlorus, Streptomyces herbaceus, Streptomyces incanus, Streptomyces pratens and Streptomyces viridis, were also studied along with six unidentified green-spored Streptomyces strains from the ARS Culture Collection. The MLSAs suggest that three of the species under study (S. bambergiensis, S. cyanoalbus and S. emeiensis) represent synonyms of other previously described species (S. prasinus, S. hirsutus and S. prasinopilosus, respectively). These relationships were confirmed through determination of in silico DNA-DNA hybridization estimates based on draft genome sequences. The five recently described species appear to be phylogenetically distinct but the unidentified strains from the ARS Culture Collection could be identified as representatives of S. hirsutus, S. prasinopilosus or S. prasinus.
Ser, Hooi-Leng; Zainal, Nurullhudda; Palanisamy, Uma Devi; Goh, Bey-Hing; Yin, Wai-Fong; Chan, Kok-Gan; Lee, Learn-Han
A novel Streptomyces, strain MUSC 26(T), was isolated from mangrove soil at Tanjung Lumpur, Malaysia. The bacterium was observed to be Gram-positive and to form grayish yellow aerial and substrate mycelium on ISP 7 agar. A polyphasic approach was used to study the taxonomy of strain MUSC 26(T), which shows a range of phylogenetic and chemotaxonomic properties consistent with those of the members of the genus Streptomyces. The cell wall peptidoglycan was determined to contain LL-diaminopimelic acid. The predominant menaquinones were identified as MK-9 (H8) and MK-9(H6). The polar lipids detected were identified as diphosphatidylglycerol, phosphatidylinositol, phosphatidylethanolamine, hydroxyphosphatidylethanolamine, phosphatidylmethylethanolamine and hydroxyphosphatidylmethylethanolamine. The predominant cellular fatty acids (>10.0 %) were identified as anteiso-C15:0 (31.4 %), iso-C16:0 (16.3 %), iso-C15:0 (13.9 %) and anteiso-C17:0 (12.6 %). The cell wall sugars were found to be galactose, glucose, mannose, ribose and rhamnose. These results suggest that MUSC 26(T) should be placed within the genus Streptomyces. Phylogenetic analysis indicated that closely related strains include Streptomyces qinglanensis 172205(T) (96.5 % sequence similarity), S. sodiiphilus YIM 80305(T) (96.5 %) and S. rimosus subsp. rimosus ATCC 10970(T) (96.4 %). DNA-DNA relatedness values between MUSC 26(T) and closely related type strains ranged from 17.0 ± 2.2 to 33.2 ± 5.3 %. Comparison of BOX-PCR fingerprints indicated MUSC 26(T) presents a unique DNA profile. The DNA G+C content was determined to be 74.6 mol%. Based on this polyphasic study of MUSC 26(T), it is concluded that this strain represents a novel species, for which the name Streptomyces gilvigriseus sp. nov. is proposed. The type strain is MUSC 26(T) (=DSMZ 42173(T) = MCCC 1K00504(T)).
Nguyen, Tuan Manh; Kim, Jaisoo
Strain T110(T) was isolated from a bamboo rhizosphere soil sample in the Republic of Korea and was found to produce antibiotics and secondary metabolites against a broad range of bacterial and fungal pathogens. It is a gram-positive actinobacterium with a straight and smooth, spore chain morphology. Morphological, physiological, and biochemical characterization suggest that T110(T) belongs to the genus Streptomyces. The predominant menaquinones of strain T110(T) were MK-9 (H6), MK-9 (H8), and MK-10 (H4). The cell wall peptidoglycan contained L L-diaminopimelic acid, glutamic acid, alanine, and glycine. Ribose and glucose were detected as whole-cell hydrolysates. The polar lipids consisted of diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, and phosphatidylinositol. The main fatty acids were anteiso-C(15:0), anteiso-C(17:0), C(16:0), and iso-C(16:0). Sequence analysis of the 16S rRNA gene (GenBank accession no. KM229361) combined with multiple alignment tools revealed that T110(T) shared the highest degree of similarity with Streptomyces albosporeus subsp. labilomyceticus NBRC 15387(T) (97.9%). However, DNA-DNA hybridization and phylogenetic analysis indicate that strain T110(T) is distinct from its most closely related species. Therefore, we conclude that strain T110(T) is a novel species of the genus Streptomyces and propose naming it Streptomyces bambusae. The type strain is T110(T) (=KEMB 9005-214(T) = KACC 18225(T) = NBRC 110903(T)).
El-Sayed E. Mostafa
Full Text Available Microbial protease represents the most important industrial enzymes, which have an active role in biotechnological processes. The objective of this study was to isolate new strain of Streptomyces that produce proteolytic enzymes with novel properties and the development of the low-cost medium. An alkaline protease producer strain NRC-15 was isolated from Egyptian soil sample. The cultural, morphological, physiological characters and chemotaxonomic evidence strongly indicated that the NRC-15 strain represents a novel species of the genus Streptomyces, hence the name Strptomyces pseudogrisiolus NRC-15. The culture conditions for higher protease production by NRC-15 were optimized with respect to carbon and nitrogen sources, metal ions, pH and temperature. Maximum protease production was obtained in the medium supplemented with 1% glucose, 1% yeast extract, 6% NaCl and 100 μmol/L of Tween 20, initial pH 9.0 at 50 °C for 96 h. The current results confirm that for this strain, a great ability to produce alkaline proteases, which supports the use of applications in industry.
Hwang, Kyu-Sang; Kim, Hyun Uk; Charusanti, Pep
Streptomyces species continue to attract attention as a source of novel medicinal compounds. Despite a long history of studies on these microorganisms, they still have many biochemical mysteries to be elucidated. Investigations of novel secondary metabolites and their biosynthetic gene clusters h...... collected in the form of databases and knowledgebases, providing predictive information and enabling one to explore experimentally unrecognized biological spaces of secondary metabolism. Herein, we review recent trends in the systems biology and biotechnology of Streptomyces species....
Full Text Available The water surface microlayer is still poorly explored, although it has been shown to contain a high density of metabolically active bacteria, often called bacterioneuston. Actinomycetes from the surface microlayer in the Trondheim fjord, Norway, have been isolated and characterized. A total of 217 isolates from two separate samples morphologically resembling the genus Streptomyces have been further investigated in this study. Antimicrobial assays showed that about 80% of the isolates exhibited antagonistic activity against nonfilamentous fungus, Gram-negative, and Gram-positive bacteria. Based on the macroscopic analyses and inhibition patterns from the antimicrobial assays, the sub-grouping of isolates was performed. Partial 16S rDNAs from the candidates from each subgroup were sequenced and phylogenetic analysis performed. 7 isolates with identical 16S rDNA sequences were further studied for the presence of PKS type I genes. Sequencing and phylogenetic analysis of the PKS gene fragments revealed that horizontal gene transfer between closely related species might have taken place. Identification of unique PKS genes in these isolates implies that dereplication can not be performed based solely on the 16S rDNA sequences. The results obtained in this study suggest that streptomycetes from the neuston population may be an interesting source for discovery of new antimicrobial agents.
Full Text Available A novel Streptomyces, strain MUSC 149T was isolated from mangrove soil. A polyphasic approach was used to study the taxonomy of MUSC 149T, which shows a range of phylogenetic and chemotaxonomic properties consistent with those of the members of the genus Streptomyces. The diamino acid of the cell wall peptidoglycan was LL-diaminopimelic acid. The predominant menaquinones were identified as MK9(H8 and MK9(H6. Phylogenetic analysis indicated that closely related strains include Streptomyces rhizophilus NBRC 108885T (99.2 % sequence similarity, Streptomyces gramineus NBRC 107863T (98.7 % and Streptomyces graminisoli NBRC 108883T (98.5 %. The DNA–DNA relatedness values between MUSC 149T and closely related type strains ranged from 12.4 ± 3.3 % to 27.3 ± 1.9 %. The DNA G + C content was determined to be 72.7 mol%. The extract of MUSC 149T exhibited strong antioxidant activity and chemical analysis reported identification of an antioxidant agent, Pyrrolo[1,2-a]pyrazine-1,4-dione, hexahydro-. These data showed that metabolites of MUSC 149T shall be useful as preventive agent against free-radical associated diseases. Based on the polyphasic study of MUSC 149T, the strain merits assignment to a novel species, for which the name Streptomyces mangrovisoli sp. nov. is proposed. The type strain is MUSC 149T (= MCCC 1K00699T = DSM 100438T.
Full Text Available Background and Purpose: Cutaneous infections arise from a homogeneous group of keratinophilic fungi, known as dermatophytes. Since these pathogenic dermatophytes are eukaryotes in nature, use of chemical antifungal agents for treatment may affect the host tissue cells. In this study, we aimed to evaluate the antifungal activity of Actinomyces species against Trichophyton mentagrophytes (abbreviated as T. mentagrophytes. The isolates were obtained from soil samples and identified by polymerase chain reaction (PCR technique. Material and Methods: In total, 100 strains of Actinomyces species were isolated from soil samples in order to determine their antagonistic activities against T. mentagrophytes in Kerman, Iran. The electron microscopic study of these isolates was performed, based on the physiological properties of these antagonists (e.g., lipase, amylase, protease and chitinase, using relevant protocols. The isolates were identified using gene 16S rDNA via PCR technique. Results: Streptomyces flavogriseus, Streptomyces zaomyceticus strain xsd08149 and Streptomyces rochei were isolated and exhibited the most significant antagonistic activities against T. mentagrophytes. Images were obtained by an electron microscope and some spores, mycelia and morphology of spore chains were identified. Molecular, morphological and biochemical characteristics of these isolates were studied, using the internal 16S rDNA gene. Active isolates of Streptomyces sequence were compared to GenBank sequences. According to nucleotide analysis, isolate D5 had maximum similarity to Streptomyces flavogriseus (99%. Conclusion: The findings of this study showed that Streptomyces isolates from soil samples could exert antifungal effects on T. mentagrophytes
Yang, P W; Li, M G; Zhao, J Y; Zhu, M Z; Shang, H; Li, J R; Cui, X L; Huang, R; Wen, M L
During the screening program for fungicides, one actinomycete strain ECO 00047 was isolated with the potential activity against fungus. According to the morphology and analysis of the nucleotide sequence of the 16S rRNA gene (1500 bp) this isolate was identified as Streptomyces diastaticus. The active compounds were separated by silica gel column chromatography, Sephadex LH-20 gel filtration and then purified by flash chromatography on C18 (20-45 microm). The chemical structure of the bioactive compounds I and II were elucidated, based on the spectroscopic data of MS, IR, UV, 1H-NMR, 13C-NMR and X-ray single crystal diffraction analysis. Compounds I and II were identical with oligomycins A and C, the macrolide antibiotics which have been known to be produced by Streptomyces diastatochromogenes, S. libani and S. avermitilis. The two compounds exhibited a strong activity against Aspergillus niger, Alternaria alternata, Botrytis cinerea and Phytophthora capsici but no activity toward bacteria. Although the two above antibiotics were known, their isolation has so far not been reported from S. diastaticus.
Cobb, Ryan E; Wang, Yajie; Zhao, Huimin
Actinobacteria, particularly those of genus Streptomyces, remain invaluable hosts for the discovery and engineering of natural products and their cognate biosynthetic pathways. However, genetic manipulation of these bacteria is often labor and time intensive. Here, we present an engineered CRISPR/Cas system for rapid multiplex genome editing of Streptomyces strains, demonstrating targeted chromosomal deletions in three different Streptomyces species and of various sizes (ranging from 20 bp to 30 kb) with efficiency ranging from 70 to 100%. The designed pCRISPomyces plasmids are amenable to assembly of spacers and editing templates via Golden Gate assembly and isothermal assembly (or traditional digestion/ligation), respectively, allowing rapid plasmid construction to target any genomic locus of interest. As such, the pCRISPomyces system represents a powerful new tool for genome editing in Streptomyces.
Nov 16, 2009 ... produce a wide range of molecules with broad spectrum of activities, that is, ... The antimicrobial activity of strain SLO-105 was evaluated on solid media by .... Fermentation, Purification and Biological Activities. Aust. J. Basic .... of Streptomyces in soil of protected forest areas from the states of. Assam and ...
Kim, Ji-Nu; Kim, Yeonbum; Jeong, Yujin; Roe, Jung-Hye; Kim, Byung-Gee; Cho, Byung-Kwan
The development of rapid and efficient genome sequencing methods has enabled us to study the evolutionary background of bacterial genetic information. Here, we present comparative genomic analysis of 17 Streptomyces species, for which the genome has been completely sequenced, using the pan-genome approach. The analysis revealed that 34,592 ortholog clusters constituted the pan-genome of these Streptomyces species, including 2,018 in the core genome, 11,743 in the dispensable genome, and 20,831 in the unique genome. The core genome was converged to a smaller number of genes than reported previously, with 3,096 gene families. Functional enrichment analysis showed that genes involved in transcription were most abundant in the Streptomyces pan-genome. Finally, we investigated core genes for the sigma factors, mycothiol biosynthesis pathway, and secondary metabolism pathways; our data showed that many genes involved in stress response and morphological differentiation were commonly expressed in Streptomyces species. Elucidation of the core genome offers a basis for understanding the functional evolution of Streptomyces species and provides insights into target selection for the construction of industrial strains.
Hwang, Kyu-Sang; Kim, Hyun Uk; Charusanti, Pep; Palsson, Bernhard Ø; Lee, Sang Yup
Streptomyces species continue to attract attention as a source of novel medicinal compounds. Despite a long history of studies on these microorganisms, they still have many biochemical mysteries to be elucidated. Investigations of novel secondary metabolites and their biosynthetic gene clusters have been more systematized with high-throughput techniques through inspections of correlations among components of the primary and secondary metabolisms at the genome scale. Moreover, up-to-date information on the genome of Streptomyces species with emphasis on their secondary metabolism has been collected in the form of databases and knowledgebases, providing predictive information and enabling one to explore experimentally unrecognized biological spaces of secondary metabolism. Herein, we review recent trends in the systems biology and biotechnology of Streptomyces species.
Reports of potato fields suppressive to common scab (CS) and of association of non-pathogenic streptomycetes with CS resistance suggest that non-pathogenic strains have potential to control or modulate CS disease. Biocontrol potential of non-pathogenic Streptomyces was examined in field experiments ...
The 10 species of Streptomyces implicated as the etiological agents in scab disease of potatoes or soft rot disease of sweet potatoes are distributed among 7 different phylogenetic clades in analyses based on 16S rRNA gene sequences, but high sequence similarity of this gene among Streptomyces speci...
Hromic, A; Kirby, R
Two mutants of Streptomyces cattleya affecting DNA repair were isolated. These mutants were analysed using spore survival curves and phage reactivation curves in the presence and absence of caffeine and arsenite. Two DNA repair systems (uvr1 and uvr2) were identified, the latter of which seems to influence genetic instability.
Autolytimycin 1 was isolated from the culture filtrate ofStreptomyces autolyticus JX-47,together with two known compounds, lebstatin 2 and 17-O-demethyl-geldanamycin 3. These compounds showed the activities of anti-HSV-I. The structure of 1 was determined by spectral analysis.
A polyphasic study was undertaken to determine the taxonomic status of a Streptomyces strain which had been isolated from a high altitude Atacama Desert soil and shown to have bioactive properties. The strain, isolate H9T, was found to have chemotaxonomic, cultural, and morphological properties that...
Full Text Available An actinomycete strain referred to as TN17 was screened for its antimicrobial activities. The taxonomic status of this strain was established. The organism was found to have morphological and chemotaxonomic characteristics typical of Streptomycetes. Based on the 16S rRNA nucleotide sequences, Streptomyces sp. TN17 was found to have a relationship with Streptomyces lilaceus, Streptomyces gobitricini and Streptomyces lavendofoliae. Combined analysis of the 16 S rRNA gene sequence (FN687757, phylogenetic analysis, fatty acids profile and physiological tests indicated that there are genotypic and phenotypic differences between TN17 and neighboring Streptomyces species’ neighbors. Therefore, TN17 is a novel species: Streptomyces sp. TN17 (=DSM 42020T=CTM50229T. A cultured extract of this strain inhibits the growth of several Gram positive and Gram negative bacteria and fungi.
Hamid Mohamed E
Full Text Available Abstract Background Drug therapy is recommended in conjunction with surgery in treatment of actinomycetoma. The specific prescription depends on the type of bacteria (actinomycetoma or fungi (eumycetoma causing the disease and their in vitro antimicrobial susceptibility. Objectives To investigate the antimicrobial susceptibility among isolates of Streptomyces spp. isolated from cases of actinomycetoma in man and animals in Sudan. Methods Streptomyces strains (n = 18 isolated from cases of actinomycetoma were tested in vitro against 15 commonly prescribed antibacterial agents using MIC agar dilution method as per standard guidelines. Results Streptomyces strains isolated from actinomycetoma fall into various phenotypic groups. All of the strains were inhibited by novobiocin (8 μg/mL, gentamycin (8, 32 μg/mL and doxycycline (32 μg/mL. Fusidic acid (64 μg/mL inhibited 94.4% of the strains; bacitracin, streptomycin, cephaloridine, clindamycin, ampicillin, rifampicin and tetracycline (64 μg/mL inhibited between 61.1 and 77.8% of the strains. All strains were found resistant to amphotericin B (64 μg/mL, penicillin (20 μg/mL and sulphamethoxazole (64 μg/mL. Conclusions Saprophytic Streptomyces spp. cause actinomycetoma in man and animal belong to separate phenotypes and have a wide range of susceptibility patterns to antimicrobial agents, which pose a lot of difficulties in selecting effective in vivo treatment for actinomycetoma.
Kashiwagi, Norimasa; Miyake, Michiru; Hirose, Shuichi; Sota, Masahiro; Ogino, Chiaki; Kondo, Akihiko
The end products from starch hydrolysis by amylases have important applications in various industries. Here, two amylases derived from two Streptomyces species that hydrolyze soluble starch from potato produced maltotriose as the primary maltooligosaccharide product. The genes, annotated as putative glycoside hydrolases, were cloned and expressed in Streptomyces lividans. These amylases displayed hydrolysis activity from pH 3 to 9.5 and were not affected by Ca(2+.) Optimal production of maltotriose was between 20 and 30 °C at pH 6.5. At the optimal temperature, both amylases produced maltotriose-rich end products rather than either maltose or maltotetraose.
Common scab of potato and other tuber/root crops, caused by a complex of soil bacteria in the genus Streptomyces, is one of the most important potato diseases in North America. Knowledge of the pathogen is fundamental to understanding and alleviating plant disease. To learn what species are found in...
Kuhn, S P; Lampel, J S; Strohl, W R
A new temperate actinophage from Streptomyces galilaeus ATCC 31133 was purified after that strain was crossed with S. peucetius ATCC 29050. Sensitive hosts became lysogenized and yielded turbid plaques of 2 to 3 mm in diameter. Host-range analysis indicated that 16 of 27 Streptomyces strains tested were sensitive to infection on solid medium. S. lividans and S. coelicolor A3(2) were among those not infected by this new actinophage. The new actinophage, designated phi SPK1, belongs to the Bradley group B morphological type, the pH optimum for infection is 6.75 to 7.0, it is not efficiently induced by mitomycin C or UV irradiation, it has a circular chromosome of 35.8 +/- 0.5 kilobase pairs in length containing overlapping (cohesive) ends, and the G+C content of its DNA was calculated from the buoyant density of 1.7240 to be 69 mol%. The DNA of phage phi SPK1 was cleaved by the restriction endonucleases ApaI, AluII, EcoRI, PvuII, and SalI, but, in all cases except that with EcoRI, treatment yielded greater than 20 restriction fragments. No sites were detected for BamHI, BclI, BglII, ClaI, HindIII, MluI, PstI, SmaI, SphI, SstI, XbaI, or XhoI.
Chellapandi, P.; Jani, Himanshu M.
Cellulase is a complex enzyme system, commercially produced by filamentous fungi under solid-state and submerged cultivation. It has wide applicability in textile, food and beverage industry for effective saccharification process. In this study, cellulolytic enzyme activity, particularly endoglucanase of 26 Streptomyces strains isolated from garden soil was examined, including two isolates selected on the basis of potential cellulolytic activity on Bennett’s agar medium. To enhance the endoglucanase formation in broth culture, different conditions including carbon and nitrogen sources, and growth conditions were tested. The maximum endoglucanase activity (11.25-11.90 U/mL) was achieved within 72-88 h in fermentation medium containing Tween-80, followed by phosphate sources. Both cellulolytic Streptomyces isolates gave almost equal quantity of enzyme in all trials. However the effect of medium ingredients on endoglucanase induction diverged with strains in some extent. PMID:24031191
Mangamuri, Ushakiranmayi; Muvva, Vijayalakshmi; Poda, Sudhakar; Naragani, Krishna; Munaganti, Rajesh Kumar; Chitturi, Bhujangarao; Yenamandra, Venkateswarlu
The strain VUK-A was isolated from a sediment sample of the Coringa mangrove ecosystem was identified as Streptomyces cheonanensis based on morphological, physiological, biochemical and molecular properties...
Two aminooligosaccharides, isovalertatins D03 (1) and D23 (2), were isolated from the culture filtrate of Streptomyces luteogriseus. Their structures were reinvestigated and revised by spectroscopic evidences including ESI multistage mass spectrometry and 2-dimensional NMR techniques.
Anné, J; Wohlleben, W; Burkardt, H J; Springer, R; Pühler, A
Several lytic and lysogenic actinophages were isolated from soil samples infected with Streptomyces cattleya and S. venezuelae. The morphologies and some biological properties of the phages, and the physico-chemical characteristics of their DNAs, were compared. Electron micrographs indicated that all the phage heads were of an icosahedral form, but head size and length of the tail varied. Two of the phages had a broad host range; the other isolates could lyse only a limited number of species. The molecular sizes of the phage DNAs were between 32.2 and 98.5 kb as estimated by electron microscopy and restriction enzyme analysis. The same study also indicated that one of the DNA species contained cohesive ends. The G + C content of the DNAs ranged between 45.1 and 74.2 mol % as estimated from melting studies. Sedimentation velocity experiments implied that several of the phage DNAs were probably heavily glycosylated or methylated. These modifications might explain the partial or slow digestion of some of the DNAs by several of the 23 restriction enzymes tested. Protoplasts of the appropriate Streptomyces strains could be efficiently transfected with phage DNA in the presence of 25% (w/v) polyethylene glycol (mol. wt 6000).
Two Streptomyces species producing antibiotic, antitumor, and anti-inflammatory compounds are widespread among intertidal macroalgae and deep-sea coral reef invertebrates from the central Cantabrian Sea.
Braña, Alfredo F; Braña, Afredo F; Fiedler, Hans-Peter; Nava, Herminio; González, Verónica; Sarmiento-Vizcaíno, Aida; Molina, Axayacatl; Acuña, José L; García, Luis A; Blanco, Gloria
Streptomycetes are widely distributed in the marine environment, although only a few studies on their associations to algae and coral ecosystems have been reported. Using a culture-dependent approach, we have isolated antibiotic-active Streptomyces species associated to diverse intertidal marine macroalgae (Phyllum Heterokontophyta, Rhodophyta, and Chlorophyta), from the central Cantabrian Sea. Two strains, with diverse antibiotic and cytotoxic activities, were found to inhabit these coastal environments, being widespread and persistent over a 3-year observation time frame. Based on 16S rRNA sequence analysis, the strains were identified as Streptomyces cyaneofuscatus M-27 and Streptomyces carnosus M-40. Similar isolates to these two strains were also associated to corals and other invertebrates from deep-sea coral reef ecosystem (Phyllum Cnidaria, Echinodermata, Arthropoda, Sipuncula, and Anelida) living up to 4.700-m depth in the submarine Avilés Canyon, thus revealing their barotolerant feature. These two strains were also found to colonize terrestrial lichens and have been repeatedly isolated from precipitations from tropospheric clouds. Compounds with antibiotic and cytotoxic activities produced by these strains were identified by high-performance liquid chromatography (HPLC) and database comparison. Antitumor compounds with antibacterial activities and members of the anthracycline family (daunomycin, cosmomycin B, galtamycin B), antifungals (maltophilins), anti-inflamatory molecules also with antituberculosis properties (lobophorins) were identified in this work. Many other compounds produced by the studied strains still remain unidentified, suggesting that Streptomyces associated to algae and coral ecosystems might represent an underexplored promising source for pharmaceutical drug discovery.
Gaurav V. Sanghvi
Full Text Available An alkaliphilic actinomycete, BCI-1, was isolated from soil samples collected from Saurashtra University campus, Gujarat. Isolated strain was identified as Streptomyces werraensis based on morphological, biochemical and phylogenetic analysis. Maximum antibiotic production was obtained in media containing sucrose 2%, Yeast extract 1.5%, and NaCl 2.5% at pH 9.0 for 7 days at 30 °C. Maximum inhibitory compound was produced at pH 9 and at 30 °C. FTIR revealed imine, amine, alkane (CC of aromatic ring and p-di substituted benzene, whereas HPLC analysis of partially purified compound and library search confirmed 95% peaks matches with erythromycin. Chloroform extracted isolated compound showed MIC values 1 μg/ml against Bacillus subtilis, ≤0.5 μg/ml against Staphylococcus aureus, ≤0.5 μg/ml against Escherichia coli and 2.0 μg/ml against Serretia GSD2 sp., which is more effective in comparison to ehtylacetate and methanol extracted compounds. The study holds significance as only few alkaliphilic actinomycetes have been explored for their antimicrobial potential.
Jenifer, John Selesteen Charles Adlin; Donio, Mariathason Birdilla Selva; Michaelbabu, Mariavincent; Vincent, Samuel Gnana Prakash; Citarasu, Thavasimuthu
Antagonistic Streptomyces spp. AJ8 was isolated and identified from the Kovalam solar salt works in India. The antimicrobial NRPS cluster gene was characterized by PCR, sequencing and predict the secondary structure analysis. The secondary metabolites will be extracted from different organic solvent extraction and studied the antibacterial, antifungal, antiviral and anticancer activities. In vitro antagonistic activity results revealed that, Streptomyces spp. AJ8 was highly antagonistic against Staphylococcus aureus, Aeromonas hydrophila WPD1 and Candida albicans. The genomic level identification revealed that, the strain was confirmed as Streptomyces spp. AJ8 and submitted the NCBI database (KC603899). The NRPS gene was generated a single gene fragment of 781 bp length (KR491940) and the database analysis revealed that, the closely related to Streptomyces spp. SAUK6068 and S. coeruleoprunus NBRC15400. The secondary metabolites extracted with ethyl acetate was effectively inhibited the bacterial and fungal growth at the ranged between 7 and 19.2 mm of zone of inhibition. The antiviral activity results revealed that, the metabolite was significantly (P < 0.001) controlled the killer shrimp virus white spot syndrome virus at the level of 85 %. The metabolite also suppressed the L929 fibroblast cancer cells at 35.7 % viability in 1000 µg treatment.
Mesquini, Josiane A; Sawaya, Alexandra C H F; López, Begonã G C; Oliveira, Valéria M; Miyasaka, Natalia R S
Atrazine is still one of the most used agricultural pesticides worldwide and it has been recognized as a major contaminant of surface and ground water. The aims of this research were to isolate an endophytic microorganism from leaves of sugarcane, evaluate its ability to degrade atrazine, and investigate the formation of metabolites. By sequencing of the 16S rRNA gene, the endophytic isolate atz2 was identified as Streptomyces sp. The reduction in atrazine concentration by Streptomyces sp. atz2 was 98 % and UHPLC-MS/MS analyses showed the appearance of an unknown metabolite observed as m/z 311. Ecotoxicity tests with an aquatic organism, Daphnia similis, confirmed that this metabolite was nontoxic. This mechanism of detoxification of atrazine is different from the ones of other free-living microorganisms that inhabit the soil or rhizosphere. The results show new aspects of atrazine detoxification, highlighting a new role of endophytic bacteria in plants.
Fan, Longjiang; Liu, Yun; Li, Zefeng; Baumann, Heike I.; Kleinschmidt, Katrin; Ye, Wanzhi; Imhoff, Johannes F.; Kleine, Michael; Cai, Daguang
Streptomyces, a branch of aerobic Gram-positive bacteria, represents the largest genus of actinobacteria. The streptomycetes are characterized by a complex secondary metabolism and produce over two-thirds of the clinically used natural antibiotics today. Here we report the draft genome sequence of a Streptomyces strain, PP-C42, isolated from the marine environment. A subset of unique genes and gene clusters for diverse secondary metabolites as well as antimicrobial peptides could be identifie...
Cho, Seung Sik; Park, Da Jeong; Simkhada, Jaya Ram; Hong, Joon Hee; Sohng, Jae Kyung; Lee, Oh Hyung; Yoo, Jin Cheol
In an attempt to isolate a biocatalyst able to catalyze biodiesel production from microbial source, Streptomyces sp. CS326 was screened from hundreds of soil isolates collected from various parts of Korea. In 16S rRNA sequence analysis, the strain showed high degree of similarity with Streptomyces xanthocidicus (99.79%); therefore, it is classified as Streptomyces sp. CS326. An extracellular lipase produced by the strain (LP326) was purified using a single step gel permeation chromatography on Sepharose CL-6B. Molecular weight of LP326 was estimated to be 17,000 Da by SDS-PAGE. The activity was optimum at 40 °C and pH 7.0 and was stable at pH 5.0-8.0 and below 50 °C. It preferred p-nitrophenyl palmitate (C16), a long chain substrate; and K (m) and V (max) for the substrate were determined to be 0.24 mM and 4.6 mM/min mg, respectively. First 10 N-terminal amino acid sequences were APDLVALQSE, which are different from so far reported lipases. LP326 catalyzed biodiesel production using methanol and various oils; therefore, the enzyme can be applicable in the field of biofuel.
Zin, Noraziah Mohamad; Baba, Mohd Shukri; Zainal-Abidin, Abu Hassan; Latip, Jalifah; Mazlan, Noor Wini; Edrada-Ebel, RuAngelie
Endophytic Streptomyces strains are potential sources for novel bioactive molecules. In this study, the diketopiperazine gancidin W (GW) was isolated from the endophytic actinobacterial genus Streptomyces, SUK10, obtained from the bark of Shorea ovalis tree, and it was tested in vivo against Plasmodium berghei PZZ1/100. GW exhibited an inhibition rate of nearly 80% at 6.25 and 3.125 μg kg−1 body weight on day four using the 4-day suppression test method on male ICR strain mice. Comparing GW at both concentrations with quinine hydrochloride and normal saline as positive and negative controls, respectively, 50% of the mice treated with 3.125 μg kg−1 body weight managed to survive for more than 11 months after infection, which almost reached the life span of normal mice. Biochemical tests of selected enzymes and proteins in blood samples of mice treated with GW were also within normal levels; in addition, no abnormalities or injuries were found on internal vital organs. These findings indicated that this isolated bioactive compound from Streptomyces SUK10 exhibits very low toxicity and is a good candidate for potential use as an antimalarial agent in an animal model. PMID:28223778
Phan Thi Hong-Thao
Full Text Available Ham Yen orange (Citrus nobilis Lour is the highly valuable commercial fruit of Vietnam. With the blooming of fruit production and farming area, this specialty crop is facing threats from several serious diseases; therefore the search for new effective biocontrollers is required to prevent the existing excessive use of fertilizers and plant protection chemicals. Endophytic actinomycetes are of great scientific interest due to their high potential of application in agriculture and pharmaceutical research. In this work, endophytic actinomycetes were isolated from a native orange species of Northeast mountainous province Tuyen Quang. Among 49 isolates obtained, the isolate TQR12-4 strongly inhibited test pathogens Colletotrichum truncatum, Geotrichum candidum, Fusarium oxysporum, and F. udum. This isolate gave comparatively high biomass yields on different substrates, for example, carboxy methyl cellulose, starch, protein, and chitin, within a wide range of temperature from 15 to 45°C and pH from 4 to 10. Sequence analysis of 16S rDNA gene showed that TQR12-4 shared 99% similarity to Streptomyces prasinopilosus; however, it slightly differed from the latter in spore morphology and hence was named as Streptomyces sp. TQR12-4. A thermostable antifungal substance of nonpeptide nature produced by Streptomyces sp. TQR12-4 had MIC against Fusarium udum of 100 μg/mL and 400 μg/mL respective to extract fractions X4 and X5.
Sakdapetsiri, Chatsuda; Fukuta, Yasuhisa; Aramsirirujiwet, Yaovapa; Shirasaka, Norifumi; Kitpreechavanich, Vichien
A total of 123 actinomycetes was isolated from 12 varieties of wild orchids and screened for potential antagonistic activity against Phytophthora, which causes black rot disease in orchids. In vitro and in vivo experimental results revealed that Streptomyces sp. strain 9X166 showed the highest antagonistic activity; its β-1,3-glucanase production ability was a key mechanism for growth inhibition of the pathogen. PCR amplification and DNA sequencing of the 16S ribosomal RNA gene allowed the identification of this strain, with high similarity (99.93%) to the novel species Streptomyces similaensis. The glucanase enzyme, purified to homogeneity by anion exchange and gel filtration chromatography, showed a specific activity of 58 U mg(-1) (a 3.9-fold increase) and yield of 6.4%. The molecular weight, as determined by SDS-PAGE and gel filtration, was approximately 99 and 80 kDa, respectively, suggesting that the enzyme was a monomer. The purified enzyme showed the highest substrate specificity to laminarin, indicating that it was β-1,3-glucanase. The hydrolyzed products of cello-oligosaccharides suggested that this enzyme was endo-type β-1,3-glucanase. Streptomyces sp. 9X166 culture filtrate, possessing β-1,3-glucanase activity, could degrade both freeze-dried and living mycelium. This is the first report on a β-1,3-glucanase-producing Streptomyces sp. that could be an effective biocontrol agent for black rot disease in orchids.
Full Text Available Actinomycete strain VY31 was isolated from agriculture soil of region Východná, Slovakia. Morphological, physiological and biochemical studies indicated that this isolate belongs to the genus Streptomyces. The 16S rRNA sequence data supported the assignment of the isolate to the genus Streptomyces rishiriensis (sequence similarity 97%. Tested isolate was able to produce melanin dark pigment and exopigments on ISP6, ISP7 and SSM+T cultivating media. The optimal pH range was from 6-8 and optimal temperature at 30 °C. The strain exhibited salt tolerance up to 5 % and utilized the carbon sources such as glucose, arabinose, xylose, inositol, mannose, fructose, rhamnose and rafinose. Using ApiZym® stripes, the highest production of enzymes was determined for phosphatase alkaline, leucinearylamidase, valinearylamidase, phosphatase acid, naphtol-AS-BI-phosphohydrolase, galactosidase and glucosidase (>40 nmol. According to ApiCoryne® results, positive reaction was confirmed in case of esculin, alkaline phosphatase, and this strain was also able to hydrolyze gelatine. Minimum Inhibitory Concentration (MIC of the purified extract of isolate was evaluated against Gram-positive bacteria Staphylococcus aureus and Enterococcus faecium, Gram-negative bacteria Escherichia coli and Pseudomonas aeruginosa and against yeast Candida albicans. On the basis of MIC results, strain VY31 had noticeable antibacterial activity against Staphylococcus aures N315 (MRSA from collection database of University Hospital in Hamburg, Germany. This isolate could be used in the development of new antibiotics for pharmaceutical purposes.
K. S. Vetrivel; K. Dharmalingam
Daunorubicin and its derivative doxorubicin are antitumour anthracycline antibiotics produced by Streptomyces peucetius. In this study we report isolation of stable mutants of S. peucetius blocked in different steps of the daunorubicin biosynthesis pathway. Mutants were screened on the basis of colony colour since producer strains are distinctively coloured on agar plates. Different mutants showed accumulation of aklaviketone, -rhodomycinone, maggiemycin or 13-dihydrocarminomycin in their culture filtrates. These results indicate that the mutations in these isolates affect steps catalysed by dnrE (mutants SPAK and SPMAG), dnrS (SPFS and SPRHO) and doxA (SPDHC) gene products.
Wu, Changsheng; Du, Chao; Gubbens, Jacob; Choi, Young Hae; van Wezel, Gilles P
Actinomycetes are a major source of antimicrobials, anticancer compounds, and other medically important products, and their genomes harbor extensive biosynthetic potential. Major challenges in the screening of these microorganisms are to activate the expression of cryptic biosynthetic gene clusters and the development of technologies for efficient dereplication of known molecules. Here we report the identification of a previously unidentified isatin-type antibiotic produced by Streptomyces sp. MBT28, following a strategy based on NMR-based metabolomics combined with the introduction of streptomycin resistance in the producer strain. NMR-guided isolation by tracking the target proton signal resulted in the characterization of 7-prenylisatin (1) with antimicrobial activity against Bacillus subtilis. The metabolite-guided genome mining of Streptomyces sp. MBT28 combined with proteomics identified a gene cluster with an indole prenyltransferase that catalyzes the conversion of tryptophan into 7-prenylisatin. This study underlines the applicability of NMR-based metabolomics in facilitating the discovery of novel antibiotics.
Rong, Xiaoying; Liu, Ning; Ruan, Jisheng; Huang, Ying
Systematics can provide a fundamental framework for understanding the relationships and diversification of organisms. Multilocus sequence analysis (MLSA) has shown great promise for an elaborate taxonomic grouping of streptomycete diversity. To evaluate the practical significance of MLSA as a valuable systematic tool for streptomycetes, we examined six endophytic Streptomyces griseus isolates and two S. griseus reference strains possessing obvious antagonistic activities and identical 16S rRNA gene sequences, using both housekeeping genes and secondary metabolic genes. All the eight strains contained PKS-I and NRPS genes, but not PKS-II genes, and showed similar diversity in both the MLSA phylogeny based on five housekeeping genes (atpD, gyrB, recA, rpoB and trpB) and fingerprinting of KS-AT genes. We also inferred a phylogeny based on concatenated amino acid sequences of representative KS-AT genes from the strains, which displayed a topology correlated well with those of housekeeping-gene MLSA and KS-AT fingerprinting. The good congruence observed between phylogenies based on the different datasets verified that the MLSA scheme provided robust resolution at intraspecific level and could predict the overall diversity of secondary metabolic potential within a Streptomyces species, despite somewhat of a discrepancy with antimicrobial data. It is therefore feasible to apply MLSA to dissecting natural diversity of streptomycetes for a better understanding of their evolution and ecology, as well as for facilitating their bioprospecting.
Eva Mathias Sosovele
Full Text Available This study was undertaken to isolate Actinomycetes from mangrove sediments of Tanzania and evaluate their potential for production of bioactive metabolites. Starch cacein agar medium was used to isolate the actinomycetes. Extraction of Actinomycetes using ethyl acetate (1:1, afforded dry extracts. The extracts were tested for antimicrobial activity and brine shrimp toxicity test. A total of three isolates (ACTN 1, ACTN 2 and ACTN 3 were obtained by using culture medium selective for Actinomycetes. Actinomycetes specific primers; S-C-Act-235-S-20 and S-C-Act-878-A-19 were used to identify two isolates as Streptomyces sp and one as actinomycetes sp. The strongest activity against bacterium (Bacillus subtillis and fungus (Candida albicans was exhibited by crude extracts of Streptomyces sp (ACTN 2 and ACTN 3. Crude extracts of all three isolates exhibited non- cytotoxic activity against brine shrimp larvae with LC50 values ranging from 250 - 446 μg/ml respectively. These results provide evidence that the mangrove sediments streptomycetes could be promising sources for antimicrobial bioactive agents.
Houssam M. Atta
Full Text Available Tunicamycin is a nucleotide antibiotic which was isolated from the fermentation broth of a Streptomyces strain No. T-4. According to the morphological, cultural, physiological and biochemical characteristics, and 16S rDNA sequence analysis, strain T-4 was identified as Streptomyces torulosus. It is active in vitro against some microbial pathogenic viz: Staphylococcus aureus, NCTC 7447; Micrococcus lutea, ATCC 9341; Bacillus subtilis, NCTC 10400; B. pumilus, NCTC; Klebsiella pneumonia, NCIMB 9111; Escherichia coli, NCTC 10416; Pseudomonas aeruginosa, ATCC 10145; Saccharomyces cerevisiae ATCC 9763; Candida albicans, IMRU 3669; Aspergillus flavus, IMI 111023; Aspergillus niger IMI 31276; Aspergillus fumigatus ATCC 16424; Fusarium oxysporum; Rhizoctonia solani; Alternaria alternata; Botrytis fabae and Penicillium chrysogenium. The production media were optimized for maximum yield of secondary metabolites. The metabolites were extracted using n-butanol (1:1, v/v at pH 7.0. The chemical structural analysis with UV, IR, and MS spectral analyses confirmed that the compound produced by Streptomyces torulosus, T-4 is tunicamycin antibiotic.
Nah, Hee-Ju; Pyeon, Hye-Rim; Kang, Seung-Hoon; Choi, Si-Sun; Kim, Eung-Soo
Actinomycetes family including Streptomyces species have been a major source for the discovery of novel natural products (NPs) in the last several decades thanks to their structural novelty, diversity and complexity. Moreover, recent genome mining approach has provided an attractive tool to screen potentially valuable NP biosynthetic gene clusters (BGCs) present in the actinomycetes genomes. Since many of these NP BGCs are silent or cryptic in the original actinomycetes, various techniques have been employed to activate these NP BGCs. Heterologous expression of BGCs has become a useful strategy to produce, reactivate, improve, and modify the pathways of NPs present at minute quantities in the original actinomycetes isolates. However, cloning and efficient overexpression of an entire NP BGC, often as large as over 100 kb, remain challenging due to the ineffectiveness of current genetic systems in manipulating large NP BGCs. This mini review describes examples of actinomycetes NP production through BGC heterologous expression systems as well as recent strategies specialized for the large-sized NP BGCs in Streptomyces heterologous hosts. PMID:28360891
Murphy, C D; Moss, S J; O'Hagan, D
Streptomyces cattleya is unusual in that it produces fluoroacetate and 4-fluorothreonine as secondary metabolites. We now report the isolation of an NAD(+)-dependent fluoroacetaldehyde dehydrogenase from S. cattleya that mediates the oxidation of fluoroacetaldehyde to fluoroacetate. This is the first enzyme to be identified that is directly involved in fluorometabolite biosynthesis. Production of the enzyme begins in late exponential growth and continues into the stationary phase. Measurement of kinetic parameters shows that the enzyme has a high affinity for fluoroacetaldehyde and glycoaldehyde, but not acetaldehyde.
Zvenigorodskiĭ, V I; Tiaglov, B V; Voeĭkova, T A
A method for chromatographic separation and quantitative determination of individual components of the antibiotic virginiamycin, produced by microbiological synthesis (Streptomyces virginiae strain 147), is described. The components, M1-2 and S1-5, were isolated from fermentation broth and identified by HPTLC and HPLC (the results obtained using the two methods correlate well with each other). Conditions of culturing of the producer and compositions of nutritive media were optimized. Using UV irradiation as a mutagenic factor, the producer was selected for increased level of synthesis of the antibiotic; this was achieved by inducing mutations that impart resistance to virginiamycin and meta-fluorophenylalanine, an analog of phenylalanine.
Zhang, Hongyu; Xie, Zeping; Lou, Tingting; Jiang, Peng
A new isobenzofuran derivative ( 1) was isolated from the marine Streptomyces sp. W007 and its structure was determined through extensive spectroscopic analyses, including 1D-NMR, 2D-NMR, and ESI-MS. The absolute configuration of compound 1 was determined by a combination of experimental analyses and comparison with reported data, including biogenetic reasoning, J-coupling analysis, NOESY, and 1H-1HCOSY. Compound 1 exhibited no cytotoxicity against human cells of gastric cancer BGC-823, lung cancer A549, and breast cancer MCF7.
Al-Dhabi, Naif Abdullah; Esmail, Galal Ali; Duraipandiyan, Veeramuthu; Valan Arasu, Mariadhas; Salem-Bekhit, Mounir M
The strain Streptomyces sp. Al-Dhabi-1 was isolated from soil sediments collected from Tharban hot spring in the southern west of Saudi Arabia using actinomycetes isolation agar and starch casein agar at 55 °C. Identification of the isolate was done according to morphological, physiological and biochemical characteristics and 16S rRNA sequence similarity as well. 16S rRNA sequence and blast analyses confirmed that the isolate belonging to the genus Streptomyces. The sequence was submitted to GenBank with accession number (KF815080). Ethyl acetate extract of Streptomyces sp. Al-Dhabi-1 showed good antimicrobial activities against tested pathogenic microbes. Minimum inhibitory concentration results showed that the best values were observed against S. agalactiae (<0.039 mg/ml) and Klebsiella pneumonia (0.125 mg/ml). Minimum inhibitory concentration of Al-Dhabi-1 against fungi; Cryptococcus neoformans (0.078 mg/ml), C. albicans (0.156 mg/ml), A. niger (0.625 mg/ml), and T. mentagrophytes (0.156 mg/ml). GC-MS analysis was used for the chemical profile of ethyl acetate extract. Benzeneacetic acid (16.02 %) and acetic acid 2-phenylethyl ester (10.35 %) were the major compounds among 31 substances found the ethyl acetate extract. According to the results of antimicrobial activity against pathogenic microbes, it is clear that the actinomycetes from hot springs with extreme environments are promising source for antimicrobial compounds.
Supong, Khomsan; Thawai, Chitti; Choowong, Wilunda; Kittiwongwattana, Chokchai; Thanaboripat, Dusanee; Laosinwattana, Chamroon; Koohakan, Prommart; Parinthawong, Nonglak; Pittayakhajonwut, Pattama
An endophytic actinomycete strain BCC72023 was isolated from rice (Oryza sativa L.) and identified as the genus Streptomyces, based on phenotypic, chemotaxonomic and 16S rRNA gene sequence analyses. The strain showed 99.80% similarity compared with Streptomyces samsunensis M1463(T). Chemical investigation led to the isolation of three macrolides, efomycins M (1), G (2) and oxohygrolidin (3), along with two polyethers, abierixin (4) and 29-O-methylabierixin (5). To our knowledge, this is the first report of efomycin M being isolated from a natural source. The compounds were identified using spectroscopic techniques and comparison with previously published data. All compounds exhibited antimalarial activity against the Plasmodium falciparum, K-1 strain, a multidrug-resistant strain, with IC50 values in a range of 1.40-5.23 μg/ml. In addition, these compounds were evaluated for biological activity against Mycobacterium tuberculosis, Bacillus cereus, Colletotrichum gloeosporioides and Colletotrichum capsici, as well as cytotoxicity against both cancerous (MCF-7, KB, NCI-H187) and non-cancerous (Vero) cells.
Full Text Available Cellulase is a complex enzyme system, commercially produced by filamentous fungi under solid-state and submerged cultivation. It has wide applicability in textile, food and beverage industry for effective saccharification process. In this study, cellulolytic enzyme activity, particularly endoglucanase of 26 Streptomyces strains isolated from garden soil was examined, including two isolates selected on the basis of potential cellulolytic activity on Bennett's agar medium. To enhance the endoglucanase formation in broth culture, different conditions including carbon and nitrogen sources, and growth conditions were tested. The maximum endoglucanase activity (11.25-11.90 U/mL was achieved within 72-88 h in fermentation medium containing Tween-80, followed by phosphate sources. Both cellulolytic Streptomyces isolates gave almost equal quantity of enzyme in all trials. However the effect of medium ingredients on endoglucanase induction diverged with strains in some extent.A celulase é um sistema enzimático complexo, produzido comercialmente a partir de fungos filamentosos através de cultivo em estádio sólido e submerso. Tem uma grande aplicação na indústria têxtil e de alimentos e bebidas no processo de sacarificação. Nesse estudo, examinou-se a atividade celulolítica, especialmente de englucanase, de 26 cepas de Streptomyces isoladas de solo, incluindo duas cepas selecionadas por sua atividade celulolítica no ágar Bennett. Para estimular a produção de englucanase em meio de cultura, diferentes condições de cultivo, incluindo fonte de carbono e nitrogênio e condições de crescimento, foram avaliadas. A atividade máxima de glucanase (11,25 a 11,90 U/mL foi obtida em 72-88h em meio de cultura contendo Tween-80, seguido por fontes de fosfato. Ambas as cepas celulolíticas de Streptomyces produziram quase a mesma quantidade de enzima em todos os experimentos. Entretanto, o efeito dos ingredientes do meio na indução da glucanase
Full Text Available Abstract Currently Streptomyces is one of the most important antibiotic producing microorganisms against several diseases. In the present study Streptomyces olivochromogenes ERINLG-261 was isolated from the soil samples of the Mudumalai hills, Western Ghats, India. Morphological, physiological, biochemical and 16S rRNA studies strongly suggested that this isolate belonged to the genus Streptomyces. ERINLG-261 showed good antimicrobial activity against different bacteria and fungi in Micromonospora fermentation medium. The active ethyl acetate extract was packed in column chromatography over silica gel which led to the isolation of 2-hydroxy-9,10-anthraquinone as the active principle. The isolated compound showed good antimicrobial activity against tested bacteria and fungi in minimum inhibitory concentration (MIC and minimum bactericidal concentration (MBC studies. The compound showed moderate in vitro antiproliferative activity against A549 and COLO320 cells. The compound was subjected to molecular docking studies for the inhibition of Topoisomerase, TtgR and Beta-lactamase enzymes which are targets for antimicrobials. Docking results of the compound showed low docking energy with these enzymes indicating its usefulness as antimicrobial agent. This is the first report of antimicrobial and antiproliferative activity of 2-hydroxy-9,10-anthraquinone isolated from Streptomyces olivochromogenes along with molecular docking studies.
Essarioui, Adil; LeBlanc, Nicholas; Kistler, Harold C; Kinkel, Linda L
Plant community characteristics impact rhizosphere Streptomyces nutrient competition and antagonistic capacities. However, the effects of Streptomyces on, and their responses to, coexisting microorganisms as a function of plant host or plant species richness have received little attention. In this work, we characterized antagonistic activities and nutrient use among Streptomyces and Fusarium from the rhizosphere of Andropogon gerardii (Ag) and Lespedeza capitata (Lc) plants growing in communities of 1 (monoculture) or 16 (polyculture) plant species. Streptomyces from monoculture were more antagonistic against Fusarium than those from polyculture. In contrast, Fusarium isolates from polyculture had greater inhibitory capacities against Streptomyces than isolates from monoculture. Although Fusarium isolates had on average greater niche widths, the collection of Streptomyces isolates in total used a greater diversity of nutrients for growth. Plant richness, but not plant host, influenced the potential for resource competition between the two taxa. Fusarium isolates had greater niche overlap with Streptomyces in monoculture than polyculture, suggesting greater potential for Fusarium to competitively challenge Streptomyces in monoculture plant communities. In contrast, Streptomyces had greater niche overlap with Fusarium in polyculture than monoculture, suggesting that Fusarium experiences greater resource competition with Streptomyces in polyculture than monoculture. These patterns of competitive and inhibitory phenotypes among Streptomyces and Fusarium populations are consistent with selection for Fusarium-antagonistic Streptomyces populations in the presence of strong Fusarium resource competition in plant monocultures. Similarly, these results suggest selection for Streptomyces-inhibitory Fusarium populations in the presence of strong Streptomyces resource competition in more diverse plant communities. Thus, landscape-scale variation in plant species richness may be
Full Text Available Thirteen strains of Streptomyces were isolated from phyllosphere of nine mangrove tree species found in Bhitarkanika mangrove ecosystem of Orissa. According to physiological, biochemical data, all 13 of the isolates were taxonomically identified to the genus Streptomyces as aurantiacus species. All strains are grayish, spirals and forming amorphous colony. Almost all utilized araginose, produced H2S, resistant towards rifampicin and penicillin, urea except few strains. However, they exhibited different extracellular activity like phosphate solubilization, lipase and L asparaginase production. This is a unique report from this mangrove ecosystem as far as Streptomyces occurrence is concerned.
Nanthini, Jayaram; Chia, Kim-Hou; Thottathil, Gincy P; Taylor, Todd D; Kondo, Shinji; Najimudin, Nazalan; Baybayan, Primo; Singh, Siddharth; Sudesh, Kumar
Streptomyces sp. strain CFMR 7, which naturally degrades rubber, was isolated from a rubber plantation. Whole genome sequencing and assembly resulted in 2 contigs with total genome size of 8.248 Mb. Two latex clearing protein (lcp) genes which are responsible for rubber degrading activities were identified.
Isolates of Nocardia cummidelens, Nocardia fluminea, Streptomyces albidoflavus, and Streptomyces luridiscabiei attributing to geosmin-related off-flavor in rainbow trout (Oncorhynchus mykiss) raised in recirculating aquaculture systems (RAS) were evaluated for the effect of temperature (10-30 degree...
Moraga, Norma Beatriz; Poma, Hugo Ramiro; Amoroso, María Julia; Rajal, Verónica Beatriz
The Salta Province - in the northwest of Argentina - is the main worldwide producer of hydroboracite and leads in exports of boron mineral and its derivatives in Latin America. In addition to the natural presence of boron compounds in the soils, there are others contaminated due to the boron mining industry. Although some bacteria are known to require boron for their growth or to be capable of storing boron, no studies have been published about Streptomyces or Lentzea genera's capacity to tolerate high boron concentrations, or about their metabolic capacities in boron contaminated environments. The results of this research show the isolation and molecular characterization of eight strains belonging to the actinobacteria phylum collected from different soils contaminated with high boron concentration in Salta state. The boron tolerance assays, which show that three of the strains were able to tolerate up 60-80 mM boron, demonstrate the potential capability of this group of bacteria to grow and maybe to remove boron from the environment. They appear to be promising, considering that these microorganisms are infrequent pathogens, are metabolically versatile and many Streptomyces can synthesize boron containing metabolites.
Tan Loh eTeng Hern
Full Text Available A Streptomyces strain, MUM256 was isolated from Tanjung Lumpur mangrove soil in Malaysia. Characterization of the strain showed that it has properties consistent with those of the members of the genus Streptomyces. In order to explore the potential bioactivities, extract of the fermented broth culture of MUM256 was prepared with organic solvent extraction method. DPPH and SOD activity were utilized to examine the antioxidant capacity and the results have revealed the potency of MUM256 in superoxide anion scavenging activity in dose-dependent manner. The cytotoxicity of MUM256 extract was determined using cell viability assay against 8 different panels of human cancer cell lines. Among all the tested cancer cells, HCT116 was the most sensitive toward the extract treatment. At the highest concentration of tested extract, the result showed 2.3, 2.0 and 1.8 folds higher inhibitory effect against HCT116, HT29 and Caco-2 respectively when compared to normal cell line. This result has demonstrated that MUM256 extract was selectively cytotoxic towards colon cancer cell lines. In order to determine the constituents responsible for its bioactivities, the extract was then subjected to chemical analysis using GC-MS. The analysis resulted in the identification of chemical constituents including phenolic and pyrrolopyrazine compounds which may responsible for antioxidant and anticancer activities observed. Based on the findings of this study, the presence of bioactive constituents in MUM256 extract could be a potential source for the development of antioxidative and chemopreventive agents.
Matselyukh, B P; Matselyukh, D Ya; Golembiovska, S L; Polishchuk, L V; Lavrinchuk, V Ya
Hyperpigmented mutants of Streptomyces globisporus 1912-Hp7 and Blakeslea trispora 18(+), 184(-) were isolated by action of hydrogen peroxide and nitrosoguanidine, correspondingly, from initial strains S. globisporus 1912-4Lcp and B. trispora 72(-), 198(+). The carotenoids of dry biomass of obtained strains, rubbed thoroughly with glass powder by a pestle in porcelain mortar were extracted by acetone and purified by TLC. Identification of the individual carotenoids was performed by means of HPLC and LC/MS spectrometry. It was shown that strain S. globisporus 1912-4Crt produced beta-carotene/lycopene (6.91/3.24 mg/L), mutants 1912-4Lcp and 1912-7Hp synthesized only lycopene (26.05 and 50.9 mg/L, respectively), and strains B. trispora 18(+) and 184(-)-beta-carotene (6.2% in dry biomass or more 2.5 g/L) without illumination in shake flasks. It is the first example of high constitutive production of the carotenoids by the representative of genus Streptomyces without photoinduction or increased synthesis of sigma factor The improved strains of B. trispora 18(+) and 184(-) can be used for biotechnological production of beta-carotene in industrial conditions.
Valli, S; Suvathi, Sugasini S; Aysha, O S; Nirmala, P; Vinoth, Kumar P; Reena, A
To evaluate the antimicrobial activity of Actinomycetes species isolated from marine environment. Twenty one strains of Actinomycetes were isolated from samples of Royapuram, Muttukadu, Mahabalipuram sea shores and Adyar estuary. Preliminary screening was done using cross-streak method against two gram-positive and eight gram-negative bacteria. The most potent strains C11 and C12 were selected from which antibacterial substances were extracted. The antibacterial activities of the extracts were performed using Kirby-Bauer disc diffusion method. Molecular identification of those isolates was done. All those twenty one isolates were active against at least one of the test organisms. Morphological characters were recorded. C11 showed activity against Staphylococcus species (13.0±0.5 mm), Vibrio harveyi (11.0±0.2 mm), Pseudomonas species (12.0±0.3 mm). C12 showed activity against Staphylococcus species (16.0±0.4 mm), Bacillus subtilis (11.0±0.2 mm), Vibrio harveyi (9.0±0.1 mm), Pseudomonas species (10.0±0.2 mm). 16S rRNA pattern strongly suggested that C11 and C12 strains were Streptomyces species. The results of the present investigation reveal that the marine Actinomycetes from coastal environment are the potent source of novel antibiotics. Isolation, characterization and study of Actinomycetes can be useful in discovery of novel species of Actinomycetes.
Valli S; Suvathi Sugasini S; Aysha OS; Nirmala P; Vinoth Kumar P; Reena A
Objective:To evaluate the antimicrobial activity of Actinomycetes species isolated from marine environment. Methods: Twenty one strains of Actinomycetes were isolated from samples of Royapuram, Muttukadu, Mahabalipuram sea shores and Adyar estuary. Preliminary screening was done using cross-streak method against two gram-positive and eight gram-negative bacteria. The most potent strains C11 and C12 were selected from which antibacterial substances were extracted. The antibacterial activities of the extracts were performed using Kirby-Bauer disc diffusion method. Molecular identification of those isolates was done. Results:All those twenty one isolates were active against at least one of the test organisms. Morphological characters were recorded. C11 showed activity against Staphylococcus species (13.0±0.5 mm), Vibrio harveyi (11.0±0.2 mm), Pseudomonas species (12.0±0.3 mm). C12 showed activity against Staphylococcus species (16.0±0.4 mm), Bacillus subtilis (11.0±0.2 mm), Vibrio harveyi (9.0±0.1 mm), Pseudomonas species (10.0±0.2 mm). 16S rRNA pattern strongly suggested that C11 and C12 strains were Streptomyces species. Conclusions: The results of the present investigation reveal that the marine Actinomycetes from coastal environment are the potent source of novel antibiotics. Isolation, characterization and study of Actinomycetes can be useful in discovery of novel species of Actinomycetes.
G. Palani Selvan
Full Text Available Objective: To investigate the antibacterial potential of the marine actinomycetes isolated from sponge samples. Methods: Thirty six marine sponge samples were collected from Palk Strait and further used for actinomycetes isolation by using serial dilution. The antibacterial activity was carried out by using cross streak assay method. Moreover, most potential strain also subjected to MIC and MBC techniques and the isolated potential strain was identified by molecular tools. Results: The maximum counts (26 x 102 CFU/g were observed in the month of May and minimum counts (1 x 102 CFU/g were noticed in April. A total of 21 actinomycetes were isolated and their antibacterial potential was assessed by using cross streak method. Among the 21 actinomycetes, the ACT-21 showed sensitivity against all the isolated fish pathogens. Further, the MIC and MBC results reveal that, the ACT-21 showed sensitivity at the concentration ranged between 500 毺 g/ mL-1 500 毺 g/mL. The phylogenetic analysis suggested that, the potential isolate ACT-21 (accession no: JF899543 showed maximum similarity index (>98% with Streptomyces sp. Conclusions: It is concluded from present study that, the crude extracts of sponge associated actinomycetes could be used as an effective antibacterial agent for the management of disease free fish culture system.
Huguet-Tapia, Jose C; Lefebure, Tristan; Badger, Jonathan H; Guan, Dongli; Pettis, Gregg S; Stanhope, Michael J; Loria, Rosemary
Streptomyces spp. are highly differentiated actinomycetes with large, linear chromosomes that encode an arsenal of biologically active molecules and catabolic enzymes. Members of this genus are well equipped for life in nutrient-limited environments and are common soil saprophytes. Out of the hundreds of species in the genus Streptomyces, a small group has evolved the ability to infect plants. The recent availability of Streptomyces genome sequences, including four genomes of pathogenic species, provided an opportunity to characterize the gene content specific to these pathogens and to study phylogenetic relationships among them. Genome sequencing, comparative genomics, and phylogenetic analysis enabled us to discriminate pathogenic from saprophytic Streptomyces strains; moreover, we calculated that the pathogen-specific genome contains 4,662 orthologs. Phylogenetic reconstruction suggested that Streptomyces scabies and S. ipomoeae share an ancestor but that their biosynthetic clusters encoding the required virulence factor thaxtomin have diverged. In contrast, S. turgidiscabies and S. acidiscabies, two relatively unrelated pathogens, possess highly similar thaxtomin biosynthesis clusters, which suggests that the acquisition of these genes was through lateral gene transfer.
Tian, Xinpeng; Zhang, Zhewen; Yang, Tingting; Chen, Meili; Li, Jie; Chen, Fei; Yang, Jin; Li, Wenjie; Zhang, Bing; Zhang, Zhang; Wu, Jiayan; Zhang, Changsheng; Long, Lijuan; Xiao, Jingfa
Over 200 genomes of streptomycete strains that were isolated from various environments are available from the NCBI. However, little is known about the characteristics that are linked to marine adaptation in marine-derived streptomycetes. The particularity and complexity of the marine environment suggest that marine streptomycetes are genetically diverse. Here, we sequenced nine strains from the Streptomyces genus that were isolated from different longitudes, latitudes, and depths of the South China Sea. Then we compared these strains to 22 NCBI downloaded streptomycete strains. Thirty-one streptomycete strains are clearly grouped into a marine-derived subgroup and multiple source subgroup-based phylogenetic tree. The phylogenetic analyses have revealed the dynamic process underlying streptomycete genome evolution, and lateral gene transfer is an important driving force during the process. Pan-genomics analyses have revealed that streptomycetes have an open pan-genome, which reflects the diversity of these streptomycetes and guarantees the species a quick and economical response to diverse environments. Functional and comparative genomics analyses indicate that the marine-derived streptomycetes subgroup possesses some common characteristics of marine adaptation. Our findings have expanded our knowledge of how ocean isolates of streptomycete strains adapt to marine environments. The availability of streptomycete genomes from the South China Sea will be beneficial for further analysis on marine streptomycetes and will enrich the South China Sea’s genetic data sources. PMID:27446038
Tian, Xinpeng; Zhang, Zhewen; Yang, Tingting; Chen, Meili; Li, Jie; Chen, Fei; Yang, Jin; Li, Wenjie; Zhang, Bing; Zhang, Zhang; Wu, Jiayan; Zhang, Changsheng; Long, Lijuan; Xiao, Jingfa
Over 200 genomes of streptomycete strains that were isolated from various environments are available from the NCBI. However, little is known about the characteristics that are linked to marine adaptation in marine-derived streptomycetes. The particularity and complexity of the marine environment suggest that marine streptomycetes are genetically diverse. Here, we sequenced nine strains from the Streptomyces genus that were isolated from different longitudes, latitudes, and depths of the South China Sea. Then we compared these strains to 22 NCBI downloaded streptomycete strains. Thirty-one streptomycete strains are clearly grouped into a marine-derived subgroup and multiple source subgroup-based phylogenetic tree. The phylogenetic analyses have revealed the dynamic process underlying streptomycete genome evolution, and lateral gene transfer is an important driving force during the process. Pan-genomics analyses have revealed that streptomycetes have an open pan-genome, which reflects the diversity of these streptomycetes and guarantees the species a quick and economical response to diverse environments. Functional and comparative genomics analyses indicate that the marine-derived streptomycetes subgroup possesses some common characteristics of marine adaptation. Our findings have expanded our knowledge of how ocean isolates of streptomycete strains adapt to marine environments. The availability of streptomycete genomes from the South China Sea will be beneficial for further analysis on marine streptomycetes and will enrich the South China Sea's genetic data sources.
She, Wenqing; Sun, Zhongfeng; Yi, Lei; Zhao, Shumiao; Liang, Yunxiang
A novel streptomycete strain, designated XY25T, was isolated from the rhizosphere soil in an alfalfa field in Jingyang, Shanxi, China. The isolate showed optimal growth at 37 °C, and was capable of growing at pH 6-10 and in the presence of 0-6 % (w/v) NaCl. Mycelia of strain XY25T appeared spiral and developed into white spore chains with long-rod spores and a smooth surface. The 16S rRNA gene sequence of XY25T was determined and was found to be highly similar to those of species of the genus Streptomyces including Streptomyces silaceus DSM 41861T (99.11 % 16S rRNA gene sequence similarity), Streptomyces flavofungini DSM 40366T (98.49 %) and Streptomyces intermedius DSM 40372T (98.43 %), all of which were used for further characterization. Each of the four streptomycetes showed distinctive patterns of carbon usage and fatty acids composition. Analysis of cellular components of strain XY25T revealed ll-diaminopimelic acid as diagnostic diamino acid and xylose as the major sugar, whereas polar lipids were determined as phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylinositol, an unknown phospholipid, two unknown phosphatidylinositol mannosides and several unknown lipids. Menaquinones were dominated by MK-9(H6) and MK-9(H8), and the main fatty acids were anteiso-C15 : 0, iso-C16 : 0 and anteiso-C17 : 0. DNA-DNA hybridization studies indicated that strain XY25T showed relatedness values of 35.2-40.42 % with the closest related species. Based on these results, strain XY25T represents a novel species of the genus Streptomyces, for which the name Streptomyces alfalfae sp. nov. is proposed. The type strain is XY25T ( = KCTC 39571T = CCTCC AA2015019T).
Foor, F; Roberts, G P; Morin, N; Snyder, L; Hwang, M; Gibbons, P H; Paradiso, M J; Stotish, R L; Ruby, C L; Wolanski, B
A temperate actinophage, TG1, was isolated from soil by growth on Streptomyces cattleya and has been shown to be potentially useful for the cloning of DNA in this organism and other streptomycetes. It forms stable lysogens by integration at a unique site on the chromosome. The phage genome consists of 41 kb of double-stranded DNA with cohesive ends. It has unique sites for ClaI, NdeI, PstI, SmaI, and XbaI. The PstI site has been shown to be in a dispensable region of the phage genome. Deletions (2 kb in length) were obtained which retain this site and should be useful for the cloning of DNA.
Nascimento, Rodrigo P; d'Avila-Levy, Claudia M; Souza, Rodrigo F; Branquinha, Marta H; Bon, Elba P S; Pereira-Jr, Nei; Coelho, Rosalie R R
Streptomyces malaysiensis AMT-3, isolated from a Brazilian cerrado soil, showed proteolytic activities detected by gelatin-sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The optimum proteinase production was obtained when using 2.5% wheat bran and 0.1% yeast extract in the culture medium, after 5 days incubation at 30 degrees C. The enzymatic complex degraded gelatin optimally at pH 7.0, and under these conditions eight proteolytic bands (four serine-proteinases and four metaloproteinases), ranging from 20 to 212 kDa, were detected on the culture supernatant filtrates. In addition, a 35-kDa proteinase was thermostable at 60 degrees C for 120 min. These results point out to the applicability of gelatin zymograms in the characterization of crude enzymatic complexes. According to our results, this enzymatic complex could be used for biotechnological applications.
Röttig, Annika; Hauschild, Philippa; Madkour, Mohamed H; Al-Ansari, Ahmed M; Almakishah, Naief H; Steinbüchel, Alexander
As oleaginous microorganisms represent an upcoming novel feedstock for the biotechnological production of lipids or lipid-derived biofuels, we searched for novel, lipid-producing strains in desert soil. This was encouraged by the hypothesis that neutral lipids represent an ideal storage compound, especially under arid conditions, as several animals are known to outlast long periods in absence of drinking water by metabolizing their body fat. Ten lipid-accumulating bacterial strains, affiliated to the genera Bacillus, Cupriavidus, Nocardia, Rhodococcus and Streptomyces, were isolated from arid desert soil due to their ability to synthesize poly(β-hydroxybutyrate), triacylglycerols or wax esters. Particularly two Streptomyces sp. strains and one Rhodococcus sp. strain accumulate significant amounts of TAG under storage conditions under optimized cultivation conditions. Rhodococcus sp. A27 and Streptomyces sp. G49 synthesized approx. 30% (w/w) fatty acids from fructose or cellobiose, respectively, while Streptomyces isolate G25 reached a cellular fatty acid content of nearly 50% (w/w) when cultivated with cellobiose. The stored triacylglycerols were composed of 30-40% branched fatty acids, such as anteiso-pentadecanoic or iso-hexadecanoic acid. To date, this represents by far the highest lipid content described for streptomycetes. A biotechnological production of such lipids using (hemi)cellulose-derived raw material could be used to obtain sustainable biodiesel with a high proportion of branched-chain fatty acids to improve its cold-flow properties and oxidative stability.
Full Text Available Background: Awareness towards the environmental pollution had made the evolution of green technology by which enzymes got special attention in industries. The enzymes replaced chemical catalysts in manufacturing various chemicals, agricultural and pharmaceutical products. Material and methods: Actinomycetes were isolated and screened for their ability to produce xylanase. For the most promising isolate, selection of media, effect of pH, temperature, metal ions, and detergents on enzyme production and activity was studied. Results: Out of 29 isolates, 22 isolates showed xylanase activity. Out of 22 xylanase producing isolate, 05 isolates were selected for secondary screening on the basis of their clear zone size. The most promising isolate PSM-3n was identified as Streptomyces albidoflavus. It produces maximum enzyme (xylanase in media Horikoshi and Ikura having carbon and nitrogen sources as oat meal and urea respectively. The optimum pH and temperature for the enzyme production was 4.0 and 45°C respectively. The enzyme activity was found maximum at temperature 50°C and enhanced in the presence of Fe3+ ions. There was a reduction in the enzyme activity in the presence of detergents like SDS, tween-20 and tween-80. The enzyme was fairly stable at 50°C for 1 h. Conclusion: The enzyme produced by the isolate PSM-3n is fairly heat stable and highly acid stable. The activity of the enzyme was increased in presence of Fe3+ ions while decreased in presence of SDS. Therefore, further studies are required for purification of xylanase for its application potential in pulp bioleaching processes and in the functional food industry.
Lu, Dandan; Ma, Zheng; Xu, Xianhao; Yu, Xiaoping
Actinomycetes have received considerable attention as biocontrol agents against fungal plant pathogens and as plant growth promoters. In this study, a total of 320 actinomycetes were isolated from various habitats in China. Among which, 77 strains have been identified as antagonistic activities against Fusarium oxysporum f. sp. cucumerinum which usually caused fusarium wilt of cucumber. Of these, isolate actinomycete M527 not only displayed broad-spectrum antifungal activity but also showed the strongest antagonistic activity against the spore germination of F. oxysporum f. sp. cucumerinum. In pot experiments, the results indicated that isolate M527 could promote the shoot growth and prevent the development of the disease on cucumber caused by F. oxysporum f. sp. cucumerinum. The control efficacy against seedling fusarium wilt of cucumber after M527 fermentation broth root-irrigation was up to 72.1% as compared to control. Based on 16S rDNA sequence analysis, the isolate M527 was identified as Streptomyces rimosus. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
Full Text Available New isolates of Streptomyces champavatii were isolated from marine sediments of the Gotland Deep (Baltic Sea, from the Urania Basin (Eastern Mediterranean, and from the Kiel Bight (Baltic Sea. The isolates produced several oligopeptidic secondary metabolites, including the new octapeptide champacyclin (1a present in all three strains. Herein, we report on the isolation, structure elucidation and determination of the absolute stereochemistry of this isoleucine/leucine (Ile/Leu = Xle rich cyclic octapeptide champacyclin (1a. As 2D nuclear magnetic resonance (NMR spectroscopy could not fully resolve the structure of (1a, additional information on sequence and configuration of stereocenters were obtained by a combination of multi stage mass spectrometry (MSn studies, amino acid analysis, partial hydrolysis and subsequent enantiomer analytics with gas chromatography positive chmical ionization/electron impact mass spectrometry (GC-PCI/EI-MS supported by comparison to reference dipeptides. Proof of the head-to-tail cyclization of (1a was accomplished by solid phase peptide synthesis (SPPS compared to an alternatively side chain cyclized derivative (2. Champacyclin (1a is likely synthesized by a non-ribosomal peptide synthetase (NRPS, because of its high content of (d-amino acids. The compound (1a showed antimicrobial activity against the phytopathogen Erwinia amylovora causing the fire blight disease of certain plants.
Full Text Available Actinomycetes are prolific producers of pharmacologically important compounds accounting for about 70% of the naturally derived antibiotics that are currently in clinical use. In this study, we report on the isolation of Streptomyces sp. strains from Mediterranean sponges, on their secondary metabolite production and on their screening for anti-infective activities. Bioassay-guided isolation and purification yielded three previously known compounds namely, cyclic depsipeptide valinomycin, indolocarbazole alkaloid staurosporine and butenolide. This is the first report of the isolation of valinomycin from a marine source. These compounds exhibited novel anti-parasitic activities specifically against Leishmania major (valinomycin IC50 < 0.11 µM; staurosporine IC50 5.30 µM and Trypanosoma brucei brucei (valinomycin IC50 0.0032 µM; staurosporine IC50 0.022 µM; butenolide IC50 31.77 µM. These results underscore the potential of marine actinomycetes to produce bioactive compounds as well as the re-evaluation of previously known compounds for novel anti-infective activities.
El-Bondkly, Ahmed M A; El-Gendy, Mervat M A; Bassyouni, Rasha H
Thirty-four endophytic marine Actinomycetes isolates were recovered from the Egyptian marine sponge Latrunculia corticata, out of them 5 isolates (14.7 %) showed red single colonies on yeast-CzAPEK plates. Isolates under the isolation code NRC50 and NRC51 were observed with the strongest red biomass. After application of protoplast fusion between NRC50 and NRC51 isolates, 26 fusants were selected and produced widely different amounts of prodigiosin-like pigments (PLPs) on different fermentation media. Among them fusant NRCF69 produced 79 and 160.4 % PLPs more than parental strains NRC50 and NRC51, respectively. According to the analysis of 16S rDNA sequence (amplified, sequenced, and submitted to GenBank under Accession no. JN232405 and JN232406, respectively), together with their morphological and biochemical characteristics, parental strains NRC50 (P1) and NRC51 (P2) were identified as Streptomyces sp. and designated as Streptomyces sp. NRC50 and Streptomyces sp. NRC51. This study describes a low cost, effective production media by using peanut seed broth, sunflower oil broth or dairy processing wastewater broth alone, or supplemented with 0.5 % mannitol that supports the production of PLPs by the Streptomyces fusant NRCF69 under study (42.03, 40.11, 36.7 and 47 g L(-1), respectively). PLPs compounds exhibited significant cytotoxic activities against three human cancer cell lines: colon cancer cell line (HCT-116), liver cancer cell line (HEPG-2) and breast cancer cell line (MCF-7) and antimycotic activity against clinical dermatophyte isolates of Trichophyton, Microsporum and Epidermophyton.
Pometto, A L; Lee, B T; Johnson, K E
Extracellular culture concentrates were prepared from Streptomyces viridosporus T7A, Streptomyces badius 252, and Streptomyces setonii 75Vi2 shake flask cultures. Ten-day-heat-treated (70 degrees C) starch-polyethylene degradable plastic films were incubated with shaking with active or inactive enzyme for 3 weeks (37 degrees C). Active enzyme illustrated changes in the films' Fourier transform infrared spectra, mechanical properties, and polyethylene molecular weight distributions. PMID:1610196
Padmanaban, Vishnu Priya; Verma, Pankaj; Venkatabaskaran, Srividhyalakshmi; Keppayan, Thirupathi; Gopal, Dharani; Sekar, Ashok Kumar; Ramalingam, Kirubagaran
Microbial-derived natural products from extreme niches such as deepsea are known to possess structural and functional novelty. With this background, the present study was designed to investigate the bioprospecting potential and systematics of a deep-sea derived piezotolerant bacterial strain NIOT-Ch-40, showing affiliation to the genus Streptomyces based on 16S RNA gene similarity. Preliminary screening for the presence of biosynthetic genes like polyketide synthase I, polyketide synthase II, non ribosomal peptide synthase, 3-amino-5-hydroxybenzoic acid synthase and spiroindimicin followed by antibacterial activity testing confirmed the presence of potent bioactivity. The secondary metabolites produced during fermentation in Streptomyces broth at 28 °C for 7 days were extracted with ethyl acetate. The extract exhibited a specific inhibitory activity against Gram-positive bacteria and was significantly effective (p Streptomyces spp. was carried out to delineate the strain NIOT-Ch-40 at a higher resolution which includes morphological, biochemical and molecular studies. Piezotolerance studies and comparison of fatty acid profiles at high pressures revealed that it could be considered as one of the taxonomic markers, especially for the strains isolated from the deep sea environments. In conclusion, the observation of comparative studies with reference strains indicated towards the strain NIOT-Ch-40 as an indigenous marine piezotolerant Streptomyces sp. with a higher probability of obtaining novel bioactive metabolites.
Briceño, Gabriela; Fuentes, María S; Rubilar, Olga; Jorquera, Milko; Tortella, Gonzalo; Palma, Graciela; Amoroso, María J; Diez, Maria Cristina
From an agricultural soil that had received continuous applications of organophosphorus pesticides, 30 actinobacteria strains were isolated. Two strains, identified as Streptomyces sp. AC1-6 and Streptomyces sp. ISP4, were selected because of their tolerance to diazinon and based on the relationship between diazinon removal and microbial growth. In liquid medium with diazinon at concentrations of 25 and 50 mg L(-1), both strains were able to remove approximately 40-50% and 70-90% of the initial diazinon after 24 and 96 h of incubation, respectively. This diazinon removal was accompanied by microbial growth of the strains, an initial pH decrease, and glucose consumption in the liquid medium. Evaluation of the diazinon removal achieved by the free actinobacteria and Streptomyces sp. AC1-6 immobilized on alginate beads revealed that the immobilized cells exhibited a 60% higher diazinon removal compared with the free cells. The reusability of the encapsulated biomass was confirmed, and a diazinon removal rate of more than 50% was obtained after the second batch. This work constitutes one of the few reports that describe Streptomyces strains as diazinon degraders. Given the high diazinon removal found, the streptomycetes exhibit suitable potential as diazinon-degrading actinobacteria for elimination of diazinon from liquid residues.
P. Saravana Kumar
Full Text Available Thirty-seven actinomycetes strains were isolated from soil samples collected from an agriculture field in Vengodu, Thiruvannamalai District, Tamil Nadu, India (latitude: 12° 54′ 0033″, North; longitude: 79° 78′ 5216″, East; elevation: 228.6/70.0 ft/m. The isolates were assessed for antagonistic activity against five Gram-positive bacteria, seven Gram-negative bacteria, and two pathogenic fungi. During the initial screening, 43% of the strains showed weak activity, 16% showed moderate activity, 5% showed good activity, and 35% showed no antagonistic activity. Among the strains tested, SCA 7 showed strong antimicrobial activity. Maximum biological activity was obtained on modified nutrient glucose agar (MNGA medium. The mycelia of SCA 7 were extracted with methanol and tested against microbial pathogens using the disc diffusion method. The crude extract was purified partially using column chromatography and assessed for antimicrobial activity. Fraction 10 showed good activity against Staphylococcus epidermidis (31.25 μg/mL and Malassezia pachydermatis (500 μg/mL and the active principle (fraction 10 was identified as 2,4-bis (1,1-dimethylethyl phenol. Based on morphological, physiological, biochemical, cultural, and molecular characteristics (16S rDNA sequencing, this strain was identified as Streptomyces sp. SCA 7. It could be used in the development of new substances for pharmaceutical or agricultural purposes.
Saravana Kumar, P; Duraipandiyan, V; Ignacimuthu, S
Thirty-seven actinomycetes strains were isolated from soil samples collected from an agriculture field in Vengodu, Thiruvannamalai District, Tamil Nadu, India (latitude: 12° 54' 0033″, North; longitude: 79° 78' 5216″, East; elevation: 228.6/70.0 ft/m). The isolates were assessed for antagonistic activity against five Gram-positive bacteria, seven Gram-negative bacteria, and two pathogenic fungi. During the initial screening, 43% of the strains showed weak activity, 16% showed moderate activity, 5% showed good activity, and 35% showed no antagonistic activity. Among the strains tested, SCA 7 showed strong antimicrobial activity. Maximum biological activity was obtained on modified nutrient glucose agar (MNGA) medium. The mycelia of SCA 7 were extracted with methanol and tested against microbial pathogens using the disc diffusion method. The crude extract was purified partially using column chromatography and assessed for antimicrobial activity. Fraction 10 showed good activity against Staphylococcus epidermidis (31.25 μg/mL) and Malassezia pachydermatis (500 μg/mL) and the active principle (fraction 10) was identified as 2,4-bis (1,1-dimethylethyl) phenol. Based on morphological, physiological, biochemical, cultural, and molecular characteristics (16S rDNA sequencing), this strain was identified as Streptomyces sp. SCA 7. It could be used in the development of new substances for pharmaceutical or agricultural purposes. Copyright © 2014. Published by Elsevier B.V.
Cox, K L; Baltz, R H
Several Streptomyces species that produce restriction endonucleases were characterized for their ability to propagate 10 different broad host range bacteriophages. Each species displayed a different pattern of plaque formation. A restrictionless mutant of S. albus G allowed plaque formation by all 10 phages, whereas the wild-type strain showed plaques with only 2 phages. DNA isolated from three of the phages was analyzed for the presence of restriction sites for Streptomyces species-encoded enzymes, and a very strong correlation was established between the failure to form plaques on Streptomyces species that produced particular restriction enzymes and the presence of the corresponding restriction sites in the phage DNA. Also, the phages that lacked restriction sites in their DNA generally formed plaques on the corresponding restriction endonuclease-producing hosts at high efficiency. The DNAs from the three phages analyzed also generally contained either many or no restriction sites for the Streptomyces species-produced enzymes, suggesting a strong evolutionary trend to either eliminate all or tolerate many restriction sites. The data indicate that restriction plays a major role in host range determination for Streptomyces phages. Analysis of bacteriophage host ranges of many other uncharacterized Streptomyces hosts has identified four relatively nonrestricting hosts, at least two of which may be suitable hosts for gene cloning. The data also suggest that several restriction systems remain to be identified in the genus Streptomyces.
Duong, Cac Thi Phung; Lee, Han-Na; Choi, Si-Sun; Lee, Sang Yup; Kim, Eung-Soo
Avermectin and its analogs are major commercial antiparasitic agents in the fields of animal health, agriculture, and human infections. Previously, comparative transcriptome analysis between the low-producer S. avermitilis ATCC31267 and the high-producer S. avermitilis ATCC31780 using a S. avermitilis whole genome chip revealed that 50 genes were overexpressed at least two-fold higher in S. avermitilis ATCC31780. To verify the biological significance of some of the transcriptomics-guided targets, five putative regulatory genes were individually cloned under the strong-andconstitutive promoter of the Streptomyces expression vector pSE34, followed by the transformation into the lowproducer S. avermitilis ATCC31267. Among the putative genes tested, three regulatory genes including SAV213, SAV3818, and SAV4023 exhibited stimulatory effects on avermectin production in S. avermitilis ATCC31267. Moreover, overexpression of SAV3818 also stimulated actinorhodin production in both S. coelicolor M145 and S. lividans TK21, implying that the SAV3818, a putative TetR-family transcriptional regulator, could be a global upregulator acting in antibiotic production in Streptomyces species.
Full Text Available The significance and frequency of marine microorganisms as producers of bioactive metabolites-a natural source of drug discovery had varied significantly during the last decades, making marine ecosystem a huge treasure trove of novel isolates and novel compounds. Among the twelve actinomycetes isolated from marine sediment sample (Lat. 17°41′962″N, Long. 83°19′633″E, amylase, protease, lipase and cellulase activities were exhibited by 8,7,4,3 isolates respectively. Five isolates exhibited l-asparaginase activity, while 5, 6, 2 isolates exhibited antibacterial, antifungal and antimicrobial activities respectively. One isolate VMS-A10 efficiently producing alpha-amylase (25.53 ± 0.50 U/mL, protease (19.26 ± 0.25 U/mL, lipase (36.25 ± 0.10 U/mL, cellulase (14.43 ± 0.513 U/mL, l-asparaginase (0.125 ± 0.004 U/mL, antimicrobial metabolites against B. subtilis (503.33 ± 5.77 U/mL, S. aureus (536.66 ± 5.77 U/mL, E. coli (533.33 ± 5.77 U/mL, P. aeruginosa (500.00 ± 10.0 U/mL, MRSA (538.33 ± 5.77 U/mL, C. albicans (353.33 ± 11.54 U/mL and A. niger (443.33 ± 15.27 U/mL was selected, identified on the basis of morphological, cultural, physiological, and biochemical properties together with 16S rDNA sequence, designated as Streptomyces parvulus strain sankarensis-A10 and sequencing product (1490 bp was deposited in the GenBank database under accession number KT906299, Culture Deposit No: NCIM-5601. Isolation and characterization of each potential actinobacteria having immense industrial and therapeutic value on an unprecedented scale from marine sediments of Visakhapatnam coast will have a burgeoning effect.
Ruanpanun, Pornthip; Laatsch, Hartmut; Tangchitsomkid, Nuchanart; Lumyong, Saisamorn
An isolate of the actinomycete, Streptomyces sp. CMU-MH021 produced secondary metabolites that inhibited egg hatch and increased juvenile mortality of the root-knot nematode Meloidogyne incognita in vitro. 16S rDNA gene sequencing showed that the isolate sequence was 99% identical to Streptomyces roseoverticillatus. The culture filtrates form different culture media were tested for nematocidal activity. The maximal activity against M. incognita was obtained by using modified basal (MB) medium. The nematicidal assay-directed fractionation of the culture broth delivered fervenulin (1) and isocoumarin (2). Fervenulin, a low molecular weight compound, shows a broad range of biological activities. However, nematicidal activity of fervenulin was not previously reported. The nematicidal activity of fervenulin (1) was assessed using the broth microdilution technique. The lowest minimum inhibitory concentrations (MICs) of the compound against egg hatch of M. incognita was 30 μg/ml and juvenile mortality of M. incognita increasing was observed at 120 μg/ml. Moreover, at the concentration of 250 μg/ml fervenulin (1) showed killing effect on second-stage nematode juveniles of M. incognita up to 100% after incubation for 96 h. Isocoumarin (2), another bioactive compound produced by Streptomyces sp. CMU-MH021, showed weak nematicidal activity with M. incognita.
Mikulík, K; Zhulanova, E; Krátký, M; Kofronová, O; Benada, O
A 4.5-kb BamHI fragment of chromosomal DNA of Streptomyces collinus containing gene ftsZ was cloned and sequenced. Upstream of ftsZ are localized genes ftsQ, murG, and ftsW, and downstream is yfiH. Gene ftsA is not adjacent to ftsZ or other genes of the cloned fragment. Protein FtsZ was isolated and characterized with respect to its binding to GTP and GTPase activity. The binding of GTP to FtsZ was Ca(2+) or Mg(2+) dependent with an optimum at 10 mM. The rate of GTP hydrolysis by FtsZ was stimulated by KCl. The presence of Ca(2+) (3-5 mM) resulted in a significant increase of GTPase activity. Higher concentrations of Ca(2+) than 5 mM had an inhibitory effect on GTPase activity. These results indicate that divalent ions (Ca(2+) or Mg(2+)) can be involved in regulation of GTP binding and hydrolysis of FtsZ. The maximum level of FtsZ was detected in aerial mycelium when spiral loops and sporulation septa were formed. FtsZ is degraded after finishing sporulation septa.
Guo, Jing; Rao, Zhiming; Yang, Taowei; Man, Zaiwei; Xu, Meijuan; Zhang, Xian
Forty-five bacterial strains that produced diffusive pigments were isolated from 40 soil samples. Maximum pigment production was from a Streptomyces kathirae strain designated SC-1. The diffused pigment was characterized by UV-visual and infrared spectroscopy, MS and (1) H nuclear magnetic resonance imaging, and was confirmed as melanin. This may be the first report of melanin production by S. kathirae. To enhance melanin production, the culture medium was optimized by conducting a series of batch fermentations in a defined medium, and the results were analysed statistically using a response surface method. The optimal culture medium comprised 3.3 g L(-1) amylodextrine, 37 g L(-1) yeast extract, 5 g L(-1) NaCl, 0.1 g L(-1) CaCl2 and 54.4 μM CuSO4 . The pH of this medium was 6.0. Under optimal conditions, the melanin concentration was maximized at 13.7 g L(-1) , c. 8.6-fold higher than obtained in suboptimal medium. To our knowledge, the results provide novel data on melanin fermentation, and identify an excellent candidate for industrial-scale microbial fermentation of melanin.
Gurusiddaiah, S; Graham, S O
The production, isolation, and some structural studies of an antifungal, antibacterial, and antiviral substance from cultures of Streptomyces hygroscopicus are described. This material, designated pantomycin, appears to be a polypeptide antibiotic with inclusion of fatty acids and carbohydrate residues. Amino acid analysis of pantomycin acid hydrolysates indicates that it contains threonine, serine, proline, glycine, alanine, valine, alloisoleucine, and an as-yet-unidentified amino acid which appears to be different from types encountered in proteinaceous materials. In addition to the aforementioned compounds, the antibiotic was shown to contain alpha-aminobutyric acid after hydrogenation. Analysis of ether extracts of the hydrolysate mixture indicated the presence of several fatty aicds; myristic, isotridecanoic, lauric, and undecylic acids. The amino and fatty acid composition of pantomycin is similar to the known antibiotic stendomycin. Pantomycin appears to also have at least one carbohydrate-like residue incorporated into its structure. The presence of carbohydrate was indicated by periodic acid-Schiff base staining of electrophoretic patterns as well as positive color formation in the phenol-sulfuric and Molisch tests, but the carbohydrate did not appear to be either a hexose or a pentose. The antibiotic, which appears to be dissociated in alcoholic solution, forms stable aggregates under aqueous conditons.
Anuradha, S; Kumar, K Shravan; Bhama, S; Kishan, V
Tuberculosis, caused by Mycobacterium tuberculosis, continues to be a serious public health problem around the world, and it urges the need for development of new antitubercular drugs. An antibiotic producing strain, Streptomyces luridus (MTCC 4402) was earlier isolated from soil by our group. In this work, the phylogenic status was established by 16S rRNA gene sequence analysis. The strain was found to be active against clinically resistant strains. The culture was grown in shake flasks in a medium containing cornsteep liquor, glucose, CaCO(3), soyabean meal and starch. Antibiotic production reached maximum at the end of 72 h. and fermentation profile was obtained. The active compound was extracted into ethyl acetate and was subjected to activity guided purification by column chromatography using silica gel, TLC and HPLC methods. The pure compound eluted at 16.7 min. by gradient elution was subjected to (1)H, (13)C NMR and mass spectral analyses. The acquired data was compared with that of natural products’ data base and found to be a known antibiotic, spiramycin. The purified compound was studied for mutagenic, cytotoxicity, antitubercular activities. It was non mutagenic at the concentration of 1000 μg/mL, non cytotoxic and active as antitubercular agent at a concentration of 64 mg/mL and was comparable to rifampicin.
Ryan F Seipke
Full Text Available Streptomyces spp. are robust producers of medicinally-, industrially- and agriculturally-important small molecules. Increased resistance to antibacterial agents and the lack of new antibiotics in the pipeline have led to a renaissance in natural product discovery. This endeavor has benefited from inexpensive high quality DNA sequencing technology, which has generated more than 140 genome sequences for taxonomic type strains and environmental Streptomyces spp. isolates. Many of the sequenced streptomycetes belong to the same species. For instance, Streptomyces albus has been isolated from diverse environmental niches and seven strains have been sequenced, consequently this species has been sequenced more than any other streptomycete, allowing valuable analyses of strain-level diversity in secondary metabolism. Bioinformatics analyses identified a total of 48 unique biosynthetic gene clusters harboured by Streptomyces albus strains. Eighteen of these gene clusters specify the core secondary metabolome of the species. Fourteen of the gene clusters are contained by one or more strain and are considered auxiliary, while 16 of the gene clusters encode the production of putative strain-specific secondary metabolites. Analysis of Streptomyces albus strains suggests that each strain of a Streptomyces species likely harbours at least one strain-specific biosynthetic gene cluster. Importantly, this implies that deep sequencing of a species will not exhaust gene cluster diversity and will continue to yield novelty.
Seipke, Ryan F
Streptomyces spp. are robust producers of medicinally-, industrially- and agriculturally-important small molecules. Increased resistance to antibacterial agents and the lack of new antibiotics in the pipeline have led to a renaissance in natural product discovery. This endeavor has benefited from inexpensive high quality DNA sequencing technology, which has generated more than 140 genome sequences for taxonomic type strains and environmental Streptomyces spp. isolates. Many of the sequenced streptomycetes belong to the same species. For instance, Streptomyces albus has been isolated from diverse environmental niches and seven strains have been sequenced, consequently this species has been sequenced more than any other streptomycete, allowing valuable analyses of strain-level diversity in secondary metabolism. Bioinformatics analyses identified a total of 48 unique biosynthetic gene clusters harboured by Streptomyces albus strains. Eighteen of these gene clusters specify the core secondary metabolome of the species. Fourteen of the gene clusters are contained by one or more strain and are considered auxiliary, while 16 of the gene clusters encode the production of putative strain-specific secondary metabolites. Analysis of Streptomyces albus strains suggests that each strain of a Streptomyces species likely harbours at least one strain-specific biosynthetic gene cluster. Importantly, this implies that deep sequencing of a species will not exhaust gene cluster diversity and will continue to yield novelty.
Full Text Available Problem statement: Soil actinomycetes particularly Streptomyces spp. showed antagonistic activity against wide range of plant pathogens. In the recent decades they have attracted high interests as biocontrol agents. Onion neck rot or gray mold caused by Botrytis allii have imposed economic post harvest damages to onion bulbs and decreased its storage durability and market value. Approach: To investigate for biocontrol means against the pathogen, antagonistic activity of 50 isolates of soil Actinomycetes were assayed through agar disk method and dual culture bioassays. Active isolates were exposed to chloroform for detection of antibiotic. Minimum Inhibitory Concentration (MIC value and solubility of active crude extract in organic solvents were determined for Streptomyces isolate No. 347 which showed a unique and stable property of inhibiting Botrytis allii. To investigate the antagonistic effect of Streptomyces isolate No. 347 on control of onion gray mold, 4 different treatments were tested by means of Tukey HSD test. Results: From the tested isolates, 13 showed anti gray mold activities. Exposure of active isolates to chloroform revealed that Streptomyces isolates No. 347, 263 and 350 retained their antifungal activities. The active metabolite(s of Streptomyces isolate No. 347 was polar, soluble in H2O but insoluble in chloroform and methanol. MIC of the crude was determined as 0.05 mg mL-1 against B. allii. Stability of the active crude in distilled water at room temperature (12-30°C was about 6 months. Statistical studies indicated that Streptomyces isolates No. 347 can decrease losses of neck rot with significant level (pConclusion: The future goals include investigation of the antifungal genes in active isolates as candidates for genetic engineering of onion for increased tolerance against B. allii.
Onaka, Hiroyasu; Mori, Yukiko; Igarashi, Yasuhiro; Furumai, Tamotsu
Natural products produced by microorganisms are important starting compounds for drug discovery. Secondary metabolites, including antibiotics, have been isolated from different Streptomyces species. The production of these metabolites depends on the culture conditions. Therefore, the development of a new culture method can facilitate the discovery of new natural products. Here, we show that mycolic acid-containing bacteria can influence the biosynthesis of cryptic natural products in Streptomyces species. The production of red pigment by Streptomyces lividans TK23 was induced by coculture with Tsukamurella pulmonis TP-B0596, which is a mycolic acid-containing bacterium. Only living cells induced this pigment production, which was not mediated by any substances. T. pulmonis could induce natural-product synthesis in other Streptomyces strains too: it altered natural-product biosynthesis in 88.4% of the Streptomyces strains isolated from soil. The other mycolic acid-containing bacteria, Rhodococcus erythropolis and Corynebacterium glutamicum, altered biosynthesis in 87.5 and 90.2% of the Streptomyces strains, respectively. The coculture broth of T. pulmonis and Streptomyces endus S-522 contained a novel antibiotic, which we named alchivemycin A. We concluded that the mycolic acid localized in the outer cell layer of the inducer bacterium influences secondary metabolism in Streptomyces, and this activity is a result of the direct interaction between the mycolic acid-containing bacteria and Streptomyces. We used these results to develop a new coculture method, called the combined-culture method, which facilitates the screening of natural products. PMID:21097597
Full Text Available Abstract Background The use of lignocellulosic materials for second generation ethanol production would give several advantages such as minimizing the conflict between land use for food and fuel production, providing less expensive raw materials than conventional agricultural feedstock, allowing lower greenhouse gas emissions than those of first generation ethanol. However, cellulosic biofuels are not produced at a competitive level yet, mainly because of the high production costs of the cellulolytic enzymes. Therefore, this study was aimed at discovering new cellulolytic microorganisms and enzymes. Results Different bacteria isolated from raw composting materials obtained from vegetable processing industry wastes were screened for their cellulolytic activity on solid medium containing carboxymethylcellulose. Four strains belonging to the actinomycetes group were selected on the basis of their phenotypic traits and cellulolytic activity on solid medium containing carboxymethylcellulose. The strain showing the highest cellulolytic activity was identified by 16S rRNA sequencing as belonging to Streptomyces genus and it was designated as Streptomyces sp. strain G12. Investigating the enzymes responsible for cellulase activity produced by Streptomyces G12 by proteomic analyses, two endoglucanases were identified. Gene coding for one of these enzymes, named CelStrep, was cloned and sequenced. Molecular analysis showed that the celstrep gene has an open reading frame encoding a protein of 379 amino acid residues, including a signal peptide of 37 amino acid residues. Comparison of deduced aminoacidic sequence to the other cellulases indicated that the enzyme CelStrep can be classified as a family 12 glycoside hydrolase. Heterologous recombinant expression of CelStrep was carried out in Escherichia coli, and the active recombinant enzyme was purified from culture supernatant and characterized. It catalyzes the hydrolysis of carboxymethylcellulose
Gopalakrishnan, Subramaniam; Vadlamudi, Srinivas; Bandikinda, Prakash; Sathya, Arumugam; Vijayabharathi, Rajendran; Rupela, Om; Kudapa, Himabindu; Katta, Krishnamohan; Varshney, Rajeev Kumar
Six actinomycetes, CAI-13, CAI-85, CAI-93, CAI-140, CAI-155 and KAI-180, isolated from six different herbal vermi-composts were characterized for in vitro plant growth-promoting (PGP) properties and further evaluated in the field for PGP activity in rice. Of the six actinomycetes, CAI-13, CAI-85, CAI-93, CAI-140 and CAI-155 produced siderophores; CAI-13, CAI-93, CAI-155 and KAI-180 produced chitinase; CAI-13, CAI-140, CAI-155 and KAI-180 produced lipase; CAI-13, CAI-93, CAI-155 and KAI-180 produced protease; and CAI-13, CAI-85, CAI-140 and CAI-155 produced ß-1-3-glucanase whereas all the six actinomycetes produced cellulase, hydrocyanic acid and indole acetic acid (IAA). The actinomycetes were able to grow in NaCl concentrations of up to 8%, at pH values between 7 and 11, temperatures between 20 and 40 °C and compatible with fungicide bavistin at field application levels. In the rice field, the actinomycetes significantly enhanced tiller numbers, panicle numbers, filled grain numbers and weight, stover yield, grain yield, total dry matter, root length, volume and dry weight over the un-inoculated control. In the rhizosphere, the actinomycetes also significantly enhanced total nitrogen, available phosphorous, % organic carbon, microbial biomass carbon and nitrogen and dehydrogenase activity over the un-inoculated control. Sequences of 16S rDNA gene of the actinomycetes matched with different Streptomyces species in BLAST analysis. Of the six actinomycetes, CAI-85 and CAI-93 were found superior over other actinomycetes in terms of PGP properties, root development and crop productivity. qRT-PCR analysis on selected plant growth promoting genes of actinomycetes revealed the up-regulation of IAA genes only in CAI-85 and CAI-93.
Ahmad, Maged S.; El-Gendy, Ahmed O.; Ahmed, Rasha R.; Hassan, Hossam M.; El-Kabbany, Hussein M.; Merdash, Ahmed G.
The occurrence of extensive antibiotics resistant bacteria increased the demands for mining out new sources of antimicrobial agents. Actinomycetes, especially Streptomyces sp. have grasped considerable attention worldwide due to production of many useful bioactive metabolites. In the present study, a total of 52 actinomycetes were isolated from agricultural soil samples in Beni-Suef, Egypt. All isolates were characterized based on colony morphology, mycelium coloration, and pigment diffusion. They were screened for their capabilities to show antimicrobial activities against different indicator microorganisms, and only 20 isolates have shown significant antimicrobial activities against at least one of the tested indicator microorganisms. The isolate AGM12-1 was active against all tested microorganisms and showed a marked antitumor activity with IC50 3.3 and 1.1 μg/ml against HCT-116 and HepG-2 cell lines respectively. It was genotypically characterized as Streptomyces sp. with the presence of PKS Π biosynthetic gene cluster. Mannitol, ammonium sulfate, pH 7, 2% inoculum size and incubation for 11 days at 30°C were the optimum conditions that used to maximize the production and hence allowed purification of one active antimicrobial compound to homogeneity using high performance liquid chromatography with a molecular mass of m/z 488.05. Nuclear magnetic resonance structural elucidation showed that this compound was a diketopiperazine derivative. PMID:28348553
Daniela A. Viana Marques
Full Text Available The selection of new microorganisms able to produce antimicrobial compounds is hoped for to reduce their production costs and the side effects caused by synthetic drugs. Clavulanic acid is a β-lactam antibiotic produced by submerged culture, which is widely used in medicine as a powerful inhibitor of β-lactamases, enzymes produced by bacteria resistant to antibiotics such penicillin and cephalosporin. The purpose of this work was to select the best clavulanic acid producer among strains of Streptomyces belonging to the Microorganism Collection of the Department of Antibiotics of the Federal University of Pernambuco (DAUFPE. Initially, the strains were studied for their capacity to inhibit the action of β-lactamases produced by Klebsiella aerogenes ATCC 15380. From these results, five strains were selected to investigate the batch kinetics of growth and clavulanic acid production in submerged culture carried out in flasks. The results were compared with the ones obtained by Streptomyces clavuligerus ATCC 27064 selected as a control strain. The best clavulanic acid producer was Streptomyces DAUFPE 3060, molecularly identified as Streptomyces variabilis, which increased the clavulanic acid production by 28% compared to the control strain. This work contributes to the enlargement of knowledge on new Streptomyces wild strains able to produce clavulanic acid by submerged culture.
Viana Marques, Daniela A; Santos-Ebinuma, Valéria de Carvalho; de Oliveira, Patrícia Maria Sobral; Lima, Gláucia Manoella de Souza; Araújo, Janete M; Lima-Filho, José L; Converti, Attilio; Pessoa-Júnior, Adalberto; Porto, Ana L F
The selection of new microorganisms able to produce antimicrobial compounds is hoped for to reduce their production costs and the side effects caused by synthetic drugs. Clavulanic acid is a β-lactam antibiotic produced by submerged culture, which is widely used in medicine as a powerful inhibitor of β-lactamases, enzymes produced by bacteria resistant to antibiotics such penicillin and cephalosporin. The purpose of this work was to select the best clavulanic acid producer among strains of Streptomyces belonging to the Microorganism Collection of the Department of Antibiotics of the Federal University of Pernambuco (DAUFPE). Initially, the strains were studied for their capacity to inhibit the action of β-lactamases produced by Klebsiella aerogenes ATCC 15380. From these results, five strains were selected to investigate the batch kinetics of growth and clavulanic acid production in submerged culture carried out in flasks. The results were compared with the ones obtained by Streptomyces clavuligerus ATCC 27064 selected as a control strain. The best clavulanic acid producer was Streptomyces DAUFPE 3060, molecularly identified as Streptomyces variabilis, which increased the clavulanic acid production by 28% compared to the control strain. This work contributes to the enlargement of knowledge on new Streptomyces wild strains able to produce clavulanic acid by submerged culture.
Oja, Terhi; San Martin Galindo, Paola; Taguchi, Takaaki; Manner, Suvi; Vuorela, Pia M; Ichinose, Koji; Metsä-Ketelä, Mikko; Fallarero, Adyary
Streptomyces bacteria are renowned for their ability to produce bioactive secondary metabolites. Recently, synthetic biology has enabled the production of intermediates and shunt products, which may have altered biological activities compared to the end products of the pathways. Here, we have evaluated the potential of recently isolated alnumycins and other closely related pyranonaphthoquinone (PNQ) polyketides against Staphylococcus aureus biofilms. The antimicrobial potency of the compounds against planktonic cells and biofilms was determined by redox dye-based viability staining, and the antibiofilm efficacy of the compounds was confirmed by viable counting. A novel antistaphylococcal polyketide, alnumycin D, was identified. Unexpectedly, the C-ribosylated pathway shunt product alnumycin D was more active against planktonic and biofilm cells than the pathway end product alnumycin A, where a ribose unit has been converted into a dioxane moiety. The evaluation of the antibiofilm potential of other alnumycins revealed that the presence of the ribose moiety in pyranose form is essential for high activity against preformed biofilms. Furthermore, the antibiofilm potential of other closely related PNQ polyketides was examined. Based on their previously reported activity against planktonic S. aureus cells, granaticin B, kalafungin, and medermycin were also selected for testing, and among them, granaticin B was found to be the most potent against preformed biofilms. The most active antibiofilm PNQs, alnumycin D and granaticin B, share several structural features that may be important for their antibiofilm activity. They are uncharged, glycosylated, and also contain a similar oxygenation pattern of the lateral naphthoquinone ring. These findings highlight the potential of antibiotic biosynthetic pathways as a source of effective antibiofilm compounds.
Noble, M; Noble, D; Fletton, R A
Streptomyces cattleya produced a new cyclopentenedione antibiotic, G2201-C [C6H6O4(I)], which is moderatley active in vitro against Gram-positive bacteria, weakly active against Gram-negative bacteria, and inactive against fungi. G2201-C is toxic to mice.
Ogawa, H; Yamashita, Y; Katahira, R; Chiba, S; Iwasaki, T; Ashizawa, T; Nakano, H
We developed a microbial prescreen using Bacillus stearothermophilus NUB3620 and bacteriophage TP-68 to detect potential antitumor compounds acting on DNA or topoisomerases. During the course of screening microbial cultures for their antibacteriophage activities, we found that Streptomyces sp. isolated from a soil sample collected in Iwakuni city, Yamaguchi prefecture, Japan, produced a new antitumor antibiotic, UCH9. UCH9 was isolated from culture broth by a combination of EtOAc extraction and column chromatography. UCH9 has a new structure related to the antitumor antibiotic chromomycins. It exhibited antimicrobial activity against Gram-positive organisms. UCH9 also showed cytotoxic activity against HeLa S3 cells with an IC50 value of 13 nM and exhibited antitumor activity in vivo against mouse leukemia P388.
Köberl, Martina; White, Richard A.; Erschen, Sabine; El-Arabi, Tarek F.; Jansson, Janet K.; Berg, Gabriele
Streptomyces sp. strain Wb2n-11, isolated from native desert soil, exhibited broad-spectrum antagonism against plant pathogenic fungi, bacteria and nematodes. The 8.2 Mb draft genome reveals genes putatively responsible for its promising biocontrol activity and genes which enable the soil bacterium to directly interact beneficially with plants.
Tsukahara, Kentaro; Toume, Kazufumi; Ito, Hanako; Ishikawa, Naoki; Ishibashi, Masami
The antibiotic β-indomycinone was isolated from Streptomyces sp. IFM11607 by cytotoxicity-guided fractionation against human gastric adenocarcinoma AGS cells, and the geometry of its δ17,18-double bond was revised from E to Z based on the coupling constant.
Zhang, Haitao; Zhang, Wei; Jin, Yan; Jin, Meifang; Yu, Xingju
A cultivation-based approach was employed to compare the culturable actinobacterial diversity associated with five marine sponge species (Craniella australiensis, Halichondria rugosa, Reniochalina sp., Sponge sp., and Stelletta tenuis). The phylogenetic affiliation of the actinobacterial isolates was assessed by 16S rDNA-RFLP analysis. A total of 181 actinobacterial strains were isolated using five different culture media (denoted as M1-M5). The type of medium exhibited significant effects on the number of actinobacteria recovered, with the highest number of isolates on M3 (63 isolates) and the lowest on M1 (12 isolates). The genera isolated were also different, with the recovery of three genera on M2 and M3, and only a single genus on M1. The number of actinobacteria isolated from the five sponge species was significantly different, with a count of 83, 36, 30, 17, and 15 isolates from S. tenuis, H. rugosa, Sponge sp., Reniochalina sp., and C. australiensis, respectively. M3 was the best isolation medium for recovery of actinobacteria from S. tenuis, H. rugosa, and Sponge sp., while no specific medium preference was observed for the recovery of actinobacteria from Reniochalina sp., and C. australiensis. The RFLP fingerprinting of 16S rDNA genes digested with HhaI revealed six different patterns, in which 16 representative 16S rDNAs were fully sequenced. Phylogenetic analysis indicated that 12 strains belong to the group Streptomyces, three strains belong to Pseudonocardia, and one strain belongs to Nocardia. Two strains C14 (from C. australiensis) and N13 (from Sponge sp.) have only 96.26% and 96.27% similarity to earlier published sequences, and are therefore potential candidates for new species. The highest diversity of three actinobacteria genera was obtained from Sponge sp., though the number of isolates was low. Two genera of actinobacteria, Streptomyces, and Pseudonocardia, were isolated from both S. tenuis and C. australiensis. Only the genus of Streptomyces
Mane, U. V.
Full Text Available The isolated Actinomycete, Streptomyces krainskii, SUK -5 was found to decolorize and degrade textile dye Reactive blue–59.This azo dye was decolorized and degraded completely by Streptomyces krainskii SUK–5 at 24 h in shaking condition in the nutrient medium at pH 8. Induction in the activity of Lignin Peroxidase,and NADH-DCIP Reductase and MR reductase represents their role in degradation .The biodegradation was monitored by TLC, UV vis spectroscopy, FTIR. and GCMS analysis. Microbial and phytotoxicity studies of the product were carried out.
Valliappan Karuppiah; Chandramohan Aarthi; Kannan Sivakumar; Lakshmanan Kannan
Objective: To enhance the pigment production by Streptomyces sp. PM4 for evaluating its anticancer activity. Methods:Response surface methodology was employed to enhance the production of red pigment from Streptomyces sp. PM4. Optimized pigment was purified and evaluated for the anticancer activity against HT1080, Hep2, HeLa and MCF7 cell lines by MTT assay. Results: Based on the response surface methodology, it could be concluded that maltose (4.06 g), peptone (7.34 g), yeast extract (4.34 g) and tyrosine (2.89 g) were required for the maximum production of pigment (1.68 g/L) by the Streptomyces sp. PM4. Optimization of the medium with the above tested features increased the pigment yield by 4.6 fold. Pigment showed the potential anticancer activity against HT1080, HEp-2, HeLa and MCF-7cell lines with the IC50 value of 18.5, 15.3, 9.6 and 8.5 respectively. Conclusions:The study revealed that the maximum amount of pigment could be produced to treat cancer.
El-Kersh, T A; Plourde, J R
Incubation of spores, washed mycelium or whole cultures of a Streptomyces sp. with chloramphenicol (I) resulted in the loss of in vitro bioactivity of the antibiotic. Gas chromatographic estimation of an appropriate extract revealed that more than 95% of the antibiotic was inactivated under the specified conditions. The spores inactivated chloramphenicol in an inorganic buffer solution, or in distilled water, without the addition of carbohydrate or external co-factor. However, addition of certain carbon sources to the spores showed a pronounced effect on the chloramphenicol transformation process and on the relative concentration of the inactivated products. Time-course studies on the spore-catalyzed chloramphenicol transformation activity showed a maximum activity at 12-hour incubation. Addition of glucose or acetate at this point maintained maximum activity. The transformation products were identified as: chloramphenicol-1-acetate (IIa); chloramphenicol-3-acetate (IIb); chloramphenicol-3-propionate (III); CHLORAMPHENICOL-O-ISOBUTYRATE (IV); chloramphenicol-3-butyrate (V); and chloramphenicol-3-isovalerate (VI), by techniques of TLC, CPC, GC, UV, IR, MS and NMR. The microbial characteristics of the isolated strain include the formation of flexuous gray aerial mycelium with smooth to rough spores, irregular in size. It is an H2S and melanin former, non-chromogenic, and was inhibited by a streptomycin-producing strain of Streptomyces griseus (Krainsky 1914) Waksman and Henrici(1948).
The ability of lignin-degrading microorganisms to attack degradable plastics was investigated in pure shake flask culture studies. The degradable plastic used in this study was produced commercially by using the Archer-Daniels-Midland POLYCLEAN masterbatch and contained pro-oxidant and 6% starch. The known lignin-degrading bacteria Streptomyces viridosporus T7A, S. badius 252, and S. setonii 75Vi2 and fungus Phanerochaete chrysosporium were used. Pro-oxidant activity was accelerated by placin...
Full Text Available Objective: To investigate the potentiality of the marine actinobacteria isolated from marine soil against fish pathogenic bacteria. Methods: In the present study, a total of 33 soil samples were collected from the Bay of Bengal, east coast region (ECR of Tamilnadu, South India. Then they were used for the isolation of actinobacteria by using conventional serial dilution technique on starch casein agar medium. The antibacterial activities of the actinobacteria were screened primarily by using cross streak plate method against fish pathogenic bacteria namely Vibrio alginolyticus, Vibrio parahaemolyticus, Vibrio cholera, Aeromonas sp. and Pseudomonas sp. The antimicrobial efficacy of the selected isolates was carried out with various organic solvents, and finally the active compound was subjected to chromatographic techniques including TLC and GC-MS. Results: Of the 82 actinobacteria isolated, 21 (26% isolates were possessed antibacterial activity against fish pathogenic bacteria. Out of 21 antibacterial isolates, the isolate ECR77 was selected for further study based on its potential activity against fish pathogenic bacteria. Of the various solvents tested, the ethyl acetate extract had good antibacterial activity against the tested bacterial pathogens. The isolate ECR77 grew well on oat meal agar medium with 2% salt level at 35 °C. GC-MS study found that the presence of bioactive compounds namely tetradecanoic acid, n-hexadecanoic acid and octadecanoic acid. The morphological, physiological, biochemical and cultural characteristics of the potential isolate were supported the identity up to generic level as Streptomyces sp. ECR77. Conclusions: The results obtained from this study concludes that the ECR soils of South India is a hot spot of novel bioactive compound producing marine actinobacteria with great pharmaceutical values.
Durairaj Thirumurugan; Ramasamy Vijayakumar
Objective: To investigate the potentiality of the marine actinobacteria isolated from marine soil against fish pathogenic bacteria.Methods:east coast region (ECR) of Tamilnadu, South India. Then they were used for the isolation of actinobacteria by using conventional serial dilution technique on starch casein agar medium. The antibacterial activities of the actinobacteria were screened primarily by using cross streak plate method against fish pathogenic bacteria namely Vibrio alginolyticus, Vibrio parahaemolyticus,Vibrio cholera, Aeromonas sp. and Pseudomonas sp. The antimicrobial efficacy of the selected isolates was carried out with various organic solvents, and finally the active compound was subjected to chromatographic techniques including TLC and GC-MS.Results:In the present study, a total of 33 soil samples were collected from the Bay of Bengal, against fish pathogenic bacteria. Out of 21 antibacterial isolates, the isolate ECR77 was selected for further study based on its potential activity against fish pathogenic bacteria. Of the various solvents tested, the ethyl acetate extract had good antibacterial activity against the tested bacterial pathogens. The isolate ECR77 grew well on oat meal agar medium with 2% salt level at 35 °C. GC-MS study found that the presence of bioactive compounds namely tetradecanoic acid,n-hexadecanoic acid and octadecanoic acid. The morphological, physiological, biochemical and cultural characteristics of the potential isolate were supported the identity up to generic level asStreptomyces sp. ECR77. Conclusions: The results obtained from this study concludes that the ECR soils of South India is a hot spot of novel bioactive compound producing marine actinobacteria with great pharmaceutical values. Of the 82 actinobacteria isolated, 21 (26%) isolates were possessed antibacterial activity.
Hee Jae Shin
Full Text Available Two new α-pyrone derivatives, violapyrones H (1 and I (2, along with known violapyrones B (3 and C (4 were isolated from the fermentation broth of a marine actinomycete Streptomyces sp. The strain was derived from a crown-of-thorns starfish, Acanthaster planci, collected from Chuuk, Federated States of Micronesia. The structures of violapyrones were elucidated by the analysis of 1D and 2D NMR and HR-ESIMS data. Violapyrones (1–4 exhibited cytotoxicity against 10 human cancer cell lines with GI50 values of 1.10–26.12 μg/mL when tested using sulforhodamine B (SRB assay. This is the first report on the cytotoxicity of violapyrones against cancer cell lines and the absolute configuration of violapyrone C.
Shah, Aabid Manzoor; Wani, Abubakar; Qazi, Parvaiz H; Rehman, Shakeel-U; Mushtaq, Saleem; Ali, Shiekh Abid; Hussain, Aehtesham; Shah, Aiyatullah; Qazi, Asif Khurshid; Makhdoomi, Ubaid Sharif; Hamid, Abid; Kumar, Ajay
The ethyl acetate extract from the fermentation broth of an actinomycete strain, identified as Streptomyces scabrisporus isolated from soil of Kashmir Himalayas - India, exhibited significant cytotoxic activity against a panel of human cancer cell lines. The active fraction subjected to column chromatography led to the isolation of pharmacologically potent anticancer compound whose structure was established to be alborixin on the basis of spectral data analysis. The compound exhibited antiproliferative activity against panel of cell lines N2a, MCF-7, MiaPaca-2, PC-3, HCT-116, MDA-MB-231, HL-60 and A-549 cells with IC50 of 9.7, 15.4, 7.2, 8.1, 3.2, 9.7, 7.5 and 11.5 μM respectively. Alborixin displayed the maximum cytotoxic activity against HCT-116 human colon carcinoma cells and therefore further studies were carried on this cell line. Alborixin decreased the clonogenic potential of HCT-116 cells in a dose dependent manner. It induced apoptotic cell death in HCT116 cells that were confirmed by Flow cytometric analysis of Annexin V/PI staining and microscopic examination of cellular morphology through DAPI-stained cells. Biochemical evidence of apoptosis came from elevating the intracellular ROS level that was accompanied by mitochondrial membrane potential loss, decreasing the expression profile of anti-apoptotic protein Bcl-2, whereas it augments cleavage of caspase-3 and PARP-1, activates caspase-8 and 9 with concomitant increase in expression of proapoptotic protein Bax in a dose dependent manner. These results indicate that alborixin obtained from Streptomyces scabrisporus IIIM55 induces apoptotic cell death in colon cancer cells HCT-116 and can be further evaluated for its potential as an anticancer agent. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.
Balachandran, C; Arun, Y; Duraipandiyan, V; Ignacimuthu, S; Balakrishna, K; Al-Dhabi, N A
Streptomyces galbus ERINLG-127 was isolated from the soil samples of the Marapalam forest, Nilgiris, India. The ethyl acetate extract was subjected to activity-guided fractionation by column chromatography over silica gel. This led to the isolation of 2,3-dihydroxy-9,10-anthraquinone as the active principle. The compound showed good antimicrobial activity against tested bacteria and fungi. The minimum inhibitory concentration values of isolated compound were 12.5 μg/mL against Pseudomonas aeruginosa, Klebsiella pneumoniae, Salmonella typhimurium, K. pneumoniae (ESBL-3971), K. pneumoniae (ESBL-3894) and Staphylococcus aureus (MRSA). The compound showed prominent cytotoxic activity in vitro against A549 lung adenocarcinoma cancer cell line. It showed 75.1 % activity at the dose of 100 μg/mL with IC50 value of 60 μg/mL. The isolated compound was subjected to molecular docking studies for the inhibition of TtgR and Topoisomerase IV enzymes which are targets for antimicrobials. Docking studies of the compound showed low docking energy indicating its usefulness as antimicrobial agent.
Full Text Available Abstract Background Whilst being closely related to the model actinomycete Streptomyces coelicolor A3(2, S. lividans 66 differs from it in several significant and phenotypically observable ways, including antibiotic production. Previous comparative gene hybridization studies investigating such differences have used low-density (one probe per gene PCR-based spotted arrays. Here we use new experimentally optimised 104,000 × 60-mer probe arrays to characterize in detail the genomic differences between wild-type S. lividans 66, a derivative industrial strain, TK24, and S. coelicolor M145. Results The high coverage and specificity (detection of three nucleotide differences of the new microarrays used has highlighted the macroscopic genomic differences between two S. lividans strains and S. coelicolor. In a series of case studies we have validated the microarray and have identified subtle changes in genomic structure which occur in the Asp-activating adenylation domains of CDA non-ribosomal peptide synthetase genes which provides evidence of gene shuffling between these domains. We also identify single nucleotide sequence inter-species differences which exist in the actinorhodin biosynthetic gene cluster. As the glyoxylate bypass is non-functional in both S. lividans strains due to the absence of the gene encoding isocitrate lyase it is likely that the ethylmalonyl-CoA pathway functions as the alternative mechanism for the assimilation of C2 compounds. Conclusions This study provides evidence for widespread genetic recombination, rather than it being focussed at 'hotspots', suggesting that the previously proposed 'archipelago model' of genomic differences between S. coelicolor and S. lividans is unduly simplistic. The two S. lividans strains investigated differ considerably in genetic complement, with TK24 lacking 175 more genes than its wild-type parent when compared to S. coelicolor. Additionally, we confirm the presence of bldB in S. lividans and
McHenney, M A; Baltz, R H
Transposon Tn5096 was inserted into a derivative of the temperature-sensitive plasmid pMT660 containing the bacteriophage FP43 pac site. The resulting plasmid, pRHB126, was transduced by FP43 into several Streptomyces species. Tn5096 transposed from pRHB126 into different sites in the genomes of Streptomyces ambofaciens, Streptomyces cinnamonensis, Streptomyces coelicolor A3(2), Streptomyces fradiae, Streptomyces griseofuscus, and Streptomyces thermotolerans.
McHenney, M A; Baltz, R H
Transposon Tn5096 was inserted into a derivative of the temperature-sensitive plasmid pMT660 containing the bacteriophage FP43 pac site. The resulting plasmid, pRHB126, was transduced by FP43 into several Streptomyces species. Tn5096 transposed from pRHB126 into different sites in the genomes of Streptomyces ambofaciens, Streptomyces cinnamonensis, Streptomyces coelicolor A3(2), Streptomyces fradiae, Streptomyces griseofuscus, and Streptomyces thermotolerans.
Full Text Available Vibrio sp. is the most serious pathogen in marine aquaculture, and the development of anti-Vibrio agents is urgently needed. However, it is extreme lack of high-throughput screening (HTS model for searching anti-Vibrio compounds. Here, we established a protein-based HTS screening model to identify agents targeting peptide deformylase (PDF of Vibrio anguillarum. To find potential anti-Vibrio compounds, crude extracts derived from marine actinomycetes were applied for screening with this model. Notably, crude extract of strain Streptomyces sp. NHF165 inhibited dramatically both on V. anguillarum PDF (VaPDF activity and V. anguillarum cell growth. And actinonin was further identified as the functional component. Anti-VaPDF and anti-V. anguillarum activities of actinonin were dose-dependent, and the IC50 values were 6.94 M and 2.85 M, respectively. To understand the resistance of V. anguillarum against actinonin, spontaneous V. anguillarum mutants with resistance against actinonin were isolated. Surprisingly, for the resistant strains, the region between 774 and 852 base pairs was found to be absent in the gene folD which produces 10-formyl-tetrahydrofolate, a donor of N-formyl to Met-tRNAfmet. When compared to the wild type strain, folD mutant showed 8 times of minimum inhibition concentration on actinonin, however, the folD complementary strain could not grow on the medium supplemented with actinonin, which suggested that folD gene mutation was mainly responsible for the actinonin resistance. To our knowledge, this is the first report showing that marine derived Streptomyces sp. could produce actinonin with anti-VaPDF activity and the resistance against actinonin by V. anguillarum is mediated by mutation in folD gene.
Wang, Xiang-Jing; Wang, Ming; Wang, Ji-Dong; Jiang, Ling; Wang, Ji-Jia; Xiang, Wen-Sheng
Bioactivity-guided fractionation of Streptomyces avermitilis NEAU1069 fermentation broth was used to isolate and determine the chemical identity of bioactive constituents with acaricidal and nemotocidal activity. The structures of novel compounds 1 and 2 were determined on the basis of spectroscopic analysis, including 1D and 2D NMR as well as HRESI-MS, ESI-MS of spectrometry analysis, UV and IR spectroscopic analyses, and comparison with data from the literature. The acaricidal activities of the isolated compounds against adult mites and mite eggs were evaluated by mortality and unhatched eggs. The nematocidal activity of the isolated compounds against Caenorhabditis elegans was calculated according to the immobilized rates against the total number of tested nematodes. The results indicated that compounds 1 and 2 exhibited potent acaricidal activity against adult mites, with a mortality of >90% at a concentration of 30 microg/mL. However, compounds 1 and 2 showed only weak acaricidal activity against mite eggs, with unhatched mite egg rates of 90% at a concentration of 10 microg/mL. These results demonstrate that compounds 1 and 2, especially compound 2, have potential as pesticides with acaricidal and nematocidal activity.
Thirumurugan, D; Vijayakumar, R
Forty marine actinobacteria were isolated from the sediments of east coast (Bay of Bengal) region of Tamilnadu, India. Morphologically distinct colonies were primarily tested against fish pathogenic bacteria such as Vibrio cholerae, V. parahaemolyticus, V. alginolyticus, Pseudomonas fluorescens and Aeromonas hydrophila by cross-streak plate method. The secondary metabolites produced by the highly potential strain cultured on starch casein broth were extracted separately with various solvents such as alcohol, ethyl acetate, methanol, petroleum ether and chloroform. The antibacterial assay of the bioactive compounds was tested against the fish pathogenic bacteria by well diffusion method. Of the various solvents used, the ethyl acetate extract of the isolate had good antibacterial activity. The potential strain was identified as Streptomyces labedae by phenotypic, 16S rRNA gene sequence and phylogenetic analysis. Purification of the biologically active compounds by column chromatography led to isolation of 27 fractions. The biologically active fraction was re-chromatographed on a silica gel column to obtain a single active compound, namely N-isopentyltridecanamide. The structure of the compounds was elucidated on the basis of ultra violet, Fourier transform infrared and nuclear magnetic resonance spectra.
Johansson, Elisabet; Reponen, Tiina; Meller, Jarek; Vesper, Stephen; Yadav, Jagjit
Both Streptomyces species and mold species have previously been isolated from moisture-damaged building materials; however, an association between these two groups of microorganisms in indoor environments is not clear. In this study we used a culture-independent method, PCR denaturing gradient gel electrophoresis (PCR-DGGE) to investigate the composition of the Streptomyces community in house dust. Twenty-three dust samples each from two sets of homes categorized as high-mold and low-mold bas...
Schaffrath, Christoph; Deng, Hai; O'Hagan, David
5'-fluorodeoxyadenosine synthase, a C-F bond-forming enzyme, has been purified from Streptomyces cattleya. The enzyme mediates a reaction between inorganic fluoride and S-adenosyl-L-methionine (SAM) to generate 5'-fluoro-5'-deoxyadenosine. The molecular weight of the monomeric protein is shown to be 32.2 kDa by electrospray mass spectrometry. The kinetic parameters for SAM (K(m) 0.42 mM, V(max) 1.28 U/mg) and fluoride ion (K(m) 8.56 mM, V(max) 1.59 U/mg) have been evaluated. Both S-adenosyl-L-homocysteine (SAH) and sinefungin were explored as inhibitors of the enzyme. SAH emerged as a potent competitive inhibitor (K(i) 29 microM) whereas sinefungin was only weakly inhibitory.
Full Text Available Purification and structure elucidation of antibiotic produced by marine actinomycetes (Streptomyces sp A11 was conducted. Production of antibiotic was carried out by liquid fermentation using yeast and peptone medium for 5 days fermentation. Purification of antibiotic was carried out by silica gel 60 (Merck, 0.063-0.200 mm column chromatography and preparative HPLC. Structure elucidation was carried out using ESI-MS, 1H NMR, 13C NMR, DEPT 13C NMR, and FTIR. This antibiotic was identified as cyclo (tyrosyl-prolyl / (C14H16N2O3. This antibiotic had biological activity to Escherichia coli ATCC 25922, Staphylococcus aureus ATCC25923, Bacillus subtilis ATCC 66923, Pseudomonas aeruginosa ATCC27853, and produced by extracellular secretion. Keywords: antibiotic, actinomycetes, purification, structure elucidation
Benouagueni, S; Ranque, S; Gacemi Kirane, D
This study aimed at describing one actinomycete strain E65 that was isolated from the water of Mellah Lake in El Kala, North-East of Algeria that produces a non-polyenic antifungal. Actinomycetes were isolated from Mellah Lake water and screened for antimicrobial activity. Antimicrobial assays were performed on ISP2 agar. The taxonomic position of the strain E65 was determined regarding phenotypic and 16S DNA sequences features. Time course of antifungal metabolites production was evaluated against Candida albicans on ISP2, ISP1 and GYEA broth. The active antifungal compound was extracted using dichloromethane and revealed by a thin layer of chromatography, chemical reagents, UV-visible and infrared spectroscopy. A total of 104 actinomycetes were isolated and screened for antimicrobial activity; 21 strains were active against Staphylococcus aureus, Escherichia coli and Candida albicans. The strain E65 showed a high in vitro activity against S. aureus and C. albicans and a good antifungal activity against a clinical C. albicans strain resistant to 5-fluorocytosine. Its 16S rRNA sequence shared 99% similarity with the Streptomyces yatensis type strain within the Streptomyces violaceusniger subclade of the Streptomyces hygroscopicus clade. It produced a non-polyenic antifungal, the IR spectrum of the antifungal extract corresponded to none of the antimicrobials compounds known to be produced by actinomycete of the S. hygroscopicus clade. The wetlands of El Kala, Algeria are a potential source of bioactive actinomycete that deserves to be explored and exploited. The Streptomyces yatensis E65 strain isolated from Mellah Lake brackish water produces a remarkable antifungal compound which original non-polyenic structure warrants further characterization. Copyright © 2014 Elsevier Masson SAS. All rights reserved.
Full Text Available In disease-suppressive soils, plants are protected from infections by specific root pathogens due to the antagonistic activities of soil and rhizosphere microorganisms. For most disease-suppressive soils, however, the microorganisms and mechanisms involved in pathogen control are largely unknown. Our recent studies identified Actinobacteria as the most dynamic phylum in a soil suppressive to the fungal root pathogen Rhizoctonia solani. Here we isolated and characterized 300 isolates of rhizospheric Actinobacteria from the Rhizoctonia-suppressive soil. Streptomyces species were the most abundant, representing approximately 70% of the isolates. Streptomyces are renowned for the production of an exceptionally large number of secondary metabolites, including volatile organic compounds (VOCs. VOC profiling of 12 representative Streptomyces isolates by SPME-GC-MS allowed a more refined phylogenetic delineation of the Streptomyces isolates than the sequencing of 16S rRNA and the house-keeping genes atpD and recA only. VOCs of several Streptomyces isolates inhibited hyphal growth of R. solani and significantly enhanced plant shoot and root biomass. Coupling of Streptomyces VOC profiles with their effects on fungal growth, pointed to VOCs potentially involved in antifungal activity. Subsequent assays with five synthetic analogues of the identified VOCs showed that methyl 2-methylpentanoate, 1,3,5-trichloro-2-methoxy benzene and the VOCs mixture have antifungal activity. In conclusion, our results point to a potential role of VOC-producing Streptomyces in disease suppressive soils and show that VOC profiling of rhizospheric Streptomyces can be used as a complementary identification tool to construct strain-specific metabolic signatures.
Cordovez, Viviane; Carrion, Victor J.; Etalo, Desalegn W.; Mumm, Roland; Zhu, Hua; van Wezel, Gilles P.; Raaijmakers, Jos M.
In disease-suppressive soils, plants are protected from infections by specific root pathogens due to the antagonistic activities of soil and rhizosphere microorganisms. For most disease-suppressive soils, however, the microorganisms and mechanisms involved in pathogen control are largely unknown. Our recent studies identified Actinobacteria as the most dynamic phylum in a soil suppressive to the fungal root pathogen Rhizoctonia solani. Here we isolated and characterized 300 isolates of rhizospheric Actinobacteria from the Rhizoctonia-suppressive soil. Streptomyces species were the most abundant, representing approximately 70% of the isolates. Streptomyces are renowned for the production of an exceptionally large number of secondary metabolites, including volatile organic compounds (VOCs). VOC profiling of 12 representative Streptomyces isolates by SPME-GC-MS allowed a more refined phylogenetic delineation of the Streptomyces isolates than the sequencing of 16S rRNA and the house-keeping genes atpD and recA only. VOCs of several Streptomyces isolates inhibited hyphal growth of R. solani and significantly enhanced plant shoot and root biomass. Coupling of Streptomyces VOC profiles with their effects on fungal growth, pointed to VOCs potentially involved in antifungal activity. Subsequent assays with five synthetic analogs of the identified VOCs showed that methyl 2-methylpentanoate, 1,3,5-trichloro-2-methoxy benzene and the VOCs mixture have antifungal activity. In conclusion, our results point to a potential role of VOC-producing Streptomyces in disease suppressive soils and show that VOC profiling of rhizospheric Streptomyces can be used as a complementary identification tool to construct strain-specific metabolic signatures. PMID:26500626
Full Text Available A total of 74 actinomycete isolates were cultivated from two marine sponges, Geodia barretti and Phakellia ventilabrum collected at the same spot at the bottom of the Trondheim fjord (Norway. Phylogenetic analyses of sponge-associated actinomycetes based on the 16S rRNA gene sequences demonstrated the presence of species belonging to the genera Streptomyces, Nocardiopsis, Rhodococcus, Pseudonocardia and Micromonospora. Most isolates required sea water for growth, suggesting them being adapted to the marine environment. Phylogenetic analysis of Streptomyces spp. revealed two isolates that originated from different sponges and had 99.7% identity in their 16S rRNA gene sequences, indicating that they represent very closely related strains. Sequencing, annotation, and analyses of the genomes of these Streptomyces isolates demonstrated that they are sister organisms closely related to terrestrial Streptomyces albus J1074. Unlike S. albus J1074, the two sponge streptomycetes grew and differentiated faster on the medium containing sea water. Comparative genomics revealed several genes presumably responsible for partial marine adaptation of these isolates. Genome mining targeted to secondary metabolite biosynthesis gene clusters identified several of those, which were not present in S. albus J1074, and likely to have been retained from a common ancestor, or acquired from other actinomycetes. Certain genes and gene clusters were shown to be differentially acquired or lost, supporting the hypothesis of divergent evolution of the two Streptomyces species in different sponge hosts.
Streptomyces scabies has been reported as the predominant cause of potato scab in Michigan. In a 2007 survey of common scab in Michigan, however, isolates were collected from a field that did not fit the description for S. scabies. Tests using species-specific PCR primers indicated isolates were S. ...
Zhu, Hong-hui; Guo, Jun; Yao, Qing; Yang, Song-zhen; Deng, Ming-rong; Li, Tai-hui
An actinomycete, designated strain GIMN4.002(T), was isolated from a tomato rhizosphere soil sample in Guangzhou, China. The strain produces white aerial mycelium and dark blue diffusible pigment on Gause's synthetic agar, and microscopic observation revealed that it produces looped chains of spiny spores. Morphological and chemotaxonomic characteristics of the strain are typical of the genus Streptomyces. Melanin was produced and antibacterial activity was detected against Gram-positive micro-organisms, such as Bacillus subtilis, Micrococcus luteus and Staphylococcus aureus. The 16S rRNA gene sequence of strain GIMN4.002(T) had highest similarity (99.4 %) to Streptomyces lincolnensis B91; however, DNA-DNA relatedness between strain GIMN4.002(T) and S. lincolnensis NBRC 13054(T) was only 32.17 %. Further, the morphological, physiological and biochemical characteristics of strain GIMN4.002(T) are distinct from S. lincolnensis and other species of the genus Streptomyces with which this strain has high 16S rRNA gene sequence similarity (98-99 %). On the basis of the physiological and molecular properties observed, it is proposed that strain GIMN4.002(T) represents a novel species of the genus Streptomyces, for which the name Streptomyces caeruleatus sp. nov. is proposed, with GIMN4.002(T) (=CCTCC M 208213(T) =NRRL B-24802(T)) as the type strain.
Streptomyces phage Nanodon is a temperate double-stranded DNA Siphoviridae belonging to cluster BD1. It was isolated from soil collected in Kilauea, HI, using Streptomyces griseus subsp. griseus as a host.
Full Text Available Objective: The clavulanic acid regulatory gene (claR is in the clavulanic acid biosyntheticgene cluster that encodes ClaR. This protein is a putative regulator of the late steps ofclavulanic acid biosynthesis. The aim of this research is the molecular cloning of claR,isolated from the Iranian strain of Streptomyces clavuligerus (S. clavuligerus.Materials and Methods: In this experimental study, two different strains of S. clavuligeruswere used (PTCC 1705 and DSM 738, of which there is no claR sequence record forstrain PTCC 1705 in all three main gene banks. The specific designed primers were subjectedto a few base modifications for introduction of the recognition sites of BamHI andClaI. The claR gene was amplified by polymerase chain reaction (PCR using DNA isolatedfrom S. clavuligerus PTCC 1705. Nested-PCR, restriction fragment length polymorphism(PCR-RFLP, and sequencing were used for molecular analysis of the claR gene.The confirmed claR was subjected to double digestion with BamHI and ClaI. The cut claRwas ligated into a pBluescript (pBs vector and transformed into E. coli.Results: The entire sequence of the isolated claR (Iranian strain was identified. Thepresence of the recombinant vector in the transformed colonies was confirmed by thecolony-PCR procedure. The correct structure of the recombinant vector, isolated from thetransformed E. coli, was confirmed using gel electrophoresis, PCR, and double digestionwith restriction enzymes.Conclusion: The constructed recombinant cassette, named pZSclaR, can be regardedas an appropriate tool for site directed mutagenesis and sub-cloning. At this time, claRhas been cloned accompanied with its precisely selected promoter so it could be used inexpression vectors. Hence the ClaR is known as a putative regulatory protein. The overproducedprotein could also be used for other related investigations, such as a mobilityshift assay.
Full Text Available A highly potent secondary metabolite producing endophytic strain, Streptomyces sp. HUST012 was isolated from the stems of the medicinal plant Dracaena cochinchinensis Lour. Strain HUST012 showed antimicrobial and antitumor activities which were significantly much higher than those of dragon's blood extracted from D. cochinchinensis Lour. On further analysis, the strain was found to produce two metabolites, SPE-B11.8 (elucidated to be a novel metabolite (Z-tridec-7-ene-1,2,13-tricarboxylic acid and SPE-B5.4 (elucidated as Actinomycin-D. The Minimum Inhibitory Concentration values of SPE-B11.8 against a set of test bacterial organisms (Methicillin-resistant Staphylococcus epidermis ATCC 35984, Methicillin-resistant Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922 and Klebsiella pneumoniae ATCC 13883 ranged between 15.63 and 62.5 µg/ml while that for SPE-B5.4 ranged between 0.04 and 2.24 µg/ml. The compound SPE-B11.8 showed cytotoxic effect at 41.63 and 29.54 µg/ml IC50 values against Hep-G2 and MCF-7 respectively, while the compound SPE-B5.4 exhibited stronger activities against them at 0.23 and 0.18 μg/ml IC50 values.
Su, Y C; Chiu, R J; Yu, N; Chang, W R
In the course of screening amylase inhibitor producing, microorganisms, a strain identified as Streptomyces nigrifaciens NTU-3314 was found to have the highest inhibitor-producing ability among the other isolated strains. This strain was aerobically cultured at 30 degrees C in a 5l jar fermentor with a working volume of 2l. The optimum cultural medium consisted of defatted soybean flake 3.0%, potato starch 4.0%, casein 0.6%, sucrose 0.6%, serine 0.02% and NaCl 0.8% (pH 7.0). With an aeration rate of 1.5 vvm, an agitation speed of 300 rpm and an inoculum of 15% seed (previously grown in seed medium 3), the highest amount of inhibitor was obtained after 24 hours of cultivation. The amylase inhibitor produced had inhibitory effects on both alpha-amylase and glucoamylase, but not on beta-amylase, alpha-glucosidase, beta-glucosidase or dextranase. It was quite stable in 0.1M phosphate buffer (pH 7.0) and nearly 100% of its activity was retained even after boiling at 100 degrees C for 20 min.
彭卫福; 吴志明; 陈未; 曾勇军; 李昆太
The research purposes of this work are to isolate and screen a strain against phytopathogenic fungi. The antagonistic strain was isolated and screened using plate confrontation culture with Rhizoctonia solani as indicator,and was further identified on the basis of morphological characteristics,physiological and biochemical characterization,and 16S rDNA sequence analysis. From 18 soil samples collected from Yunnan,Guangdong,Anhui,Hubei,and Jiangxi Provinces of China,the isolated actinomycete strain C2 presented a strong inhibitory effect on R. solani,and it was preliminarily identified as Streptomyces triostinicus,designated as Streptomyces strain C2 here. The bioassay results showed that Streptomyces strain C2 possessed a broad-spectrum inhibitory effect on a range of plant fungi such as R. solani,Pyricularia grisea,Colletotrichum gloeosporioides,Fusarium oxysporum f. sp. niveum,and Penicillium citrinum,and the inhibition percentage was 49.78%,56.62%,80.96%,18.59%,and 94.10%,respectively. Conclusively,Streptomyces strain C2 was an antagonistic strain with broad-spectrum in suppressing the growth of various plant fungal pathogens.%旨在分离筛选获得对植物病原真菌具有抑制作用的拮抗菌。以水稻纹枯病菌为指示菌，采用平板对峙培养法进行拮抗菌的分离筛选；根据菌体形态学观察、生理生化特征以及16S rDNA 序列分析，对拮抗菌进行菌种鉴定。结果显示，从采集自云南、广东、安徽、湖北、江西等地的18个土样中分离筛选到一株对水稻纹枯病菌具有良好抑制效果的菌株 C2，该菌株初步鉴定为 Streptomyces triostinicus，并暂命名为链霉菌 C2；进一步测定链霉菌 C2的抗菌谱发现，它对水稻纹枯病菌、水稻稻瘟病菌、葡萄炭疽病菌、西瓜枯萎病菌和橘青霉等多种植物病原真菌均具有良好的抑制作用，其抑菌率分别为49.78%、56.62%、80.96%、18.59%和94.10%。链霉菌 C2是一株植物病原真菌广谱拮抗菌。
Du, Hai; Lu, Hu; Xu, Yan; Du, Xiaowei
Diverse Streptomyces species act as geosmin producers in the Chinese liquor-making process. In this paper, the ecology of these Streptomyces species was analyzed using denaturing gradient gel electrophoresis (DGGE) of amplified Actinobacteria -specified rDNA. The result showed that Streptomyces were widely distributed during Daqu incubation, and multiple processing, geographic, and climate factors can affect their distribution and diversity. The genes associated with geosmin production were characterized in four geosmin-producing Streptomyces strains, all of which were isolated from geosmin-contaminated Daqu. On the basis of this information, a real-time PCR method was developed, enabling the detection of traces of Streptomyces in complex solid-state matrices. The primer was targeted at the gene coding for geosmin synthase (geoA). The real-time PCR method was found to be specific for geosmin-producing Streptomyces and did not show any cross-reactivity with geosmin-negative isolates, which are frequently present in the Chinese liquor-brewing process. Quantification of geoA in the Chinese liquor-making process could permit the monitoring of the level of geosmin producers prior to the occurrence of geosmin production. Comparison of the qPCR results based on the gene encoding geosmin synthase and Actinobacteria-specified rDNA showed that about 1-10% of the Actinobacteria carry the geosmin synthesis gene.
Lakshmipathy Deepika; Krishnan Kannabiran
Actinomycetes were isolated from the marine soil samples collected at the Ennore saltpan and were screened for biosurfactant and heavy metal resistance activity. Biosurfactant activity was evaluated by haemolysis, drop collapsing test and lipase production. Similarly heavy metal resistance was determined by tube method and agar diffusion method. Among them, two actinomycetes isolates VITDDK1 and VITDDK2 exhibited significant biosurfactant and heavy metal resistance activity. Based on the Hide...
Hojati, Zohreh; Salehi, Zahra; Motovali-Bashi, Majid; Korbekandi, Hasan; Jami, Saeed
Objective: The clavulanic acid regulatory gene (claR) is in the clavulanic acid biosynthetic gene cluster that encodes ClaR. This protein is a putative regulator of the late steps of clavulanic acid biosynthesis. The aim of this research is the molecular cloning of claR, isolated from the Iranian strain of Streptomyces clavuligerus (S. clavuligerus). Materials and Methods: In this experimental study, two different strains of S. clavuligerus were used (PTCC 1705 and DSM 738), of which there is no claR sequence record for strain PTCC 1705 in all three main gene banks. The specific designed primers were subjected to a few base modifications for introduction of the recognition sites of BamHI and ClaI. The claR gene was amplified by polymerase chain reaction (PCR) using DNA isolated from S. clavuligerus PTCC 1705. Nested-PCR, restriction fragment length polymorphism (PCR-RFLP), and sequencing were used for molecular analysis of the claR gene. The confirmed claR was subjected to double digestion with BamHI and ClaI. The cut claR was ligated into a pBluescript (pBs) vector and transformed into E. coli. Results: The entire sequence of the isolated claR (Iranian strain) was identified. The presence of the recombinant vector in the transformed colonies was confirmed by the colony-PCR procedure. The correct structure of the recombinant vector, isolated from the transformed E. coli, was confirmed using gel electrophoresis, PCR, and double digestion with restriction enzymes. Conclusion: The constructed recombinant cassette, named pZSclaR, can be regarded as an appropriate tool for site directed mutagenesis and sub-cloning. At this time, claR has been cloned accompanied with its precisely selected promoter so it could be used in expression vectors. Hence the ClaR is known as a putative regulatory protein. The overproduced protein could also be used for other related investigations, such as a mobility shift assay. PMID:23508694
Yan, Yan; Zhao, Xin; Zhang, Shun-Cang; Liu, Yan; Liang, Zong-Suo
Our previous research indicated that the Streptomyces pactum Act12 (Act12) had a certain promotional effect on tanshinone accumulation and up-regulated the expression of genes 3-hydroxy-3-methyglutaryl-CoA reductase (HMGR) and 1-deoxy-d-xylulose-5-phosphate reductoisomerase (DXR) in Salvia miltiorrhiza hairy roots. This study focuses on the roles of reactive oxygen species in S. pactum Act12-induced tanshinone production in S. miltiorrhiza hairy roots. The 4% Act12, 4% Act12 + CAT and 4% Act12 + SOD were added to S. miltiorrhiza hairy root and subcultured for 21 days, the dry weight, contents of reactive oxygen species, contents of tanshinones and expression of HMGR and DXR were determined at different harvest-time. The generation of reactive oxygen species (ROS) in S. miltiorrhiza hairy roots was triggered by 4% Act12 treatment. The relative expressions of genes HMGR and DXR in 4% Act12 treatment were 32.4 and 4.8-fold higher than those in the control. And the total tanshinone in the hairy roots was 10.2 times higher than that of the control. The CAT and SOD could significantly inhibit the ROS accumulation and relative expressions of genes HMGR and DXR in 4% Act12 treatment, which induced the total tanshinone content was decreased by 74.6% comparing with the 4% Act12 treatment. ROS mediated Act12-induced tanshinone production. The Act12 may be via the ROS signal channel to activate the tanshinone biosynthesis pathways. Thereby the tanshinon content in hairy roots was increased.
Schrader, Kevin K; Harries, Marcuslene D; Page, Phaedra N
Isolates of Nocardia cummidelens, Nocard ia fluminea, Streptomyces albidoflavus, and Streptomyces luridiscabiei attributed as the cause of "earthy-musty" off-flavor in rainbow trout (Oncorhynchus mykiss) raised in recirculating aquaculture systems (RAS) were evaluated for the effect of temperature (10-30 °C) on biomass, geosmin, and 2-methylisoborneol (MIB) production and cellular activity. Cultures of these isolates were monitored over 7 days by measuring culture dry weight, geosmin, and MIB production using solid-phase microextraction-gas chromatography-mass spectrometry (SPME-GC-MS), and ATP production via a luminometer. Compared to the other isolates, S. luridiscabiei had significantly (P geosmin (69,976 ± 15,733 ng/L) at 15 °C. At 25 °C and 30 °C, S. albidoflavus produced significantly (P geosmin (182,074 ± 60,272 ng/L and 399,991 ± 102,262 ng/L, respectively). All isolates produced MIB at 15 °C, but S. luridiscabiei produced significantly (P geosmin and MIB in the RAS.
Mavituna, Ferda; Luti, Khalid Jaber Kadhum; Gu, Lixing
The aim of this work was to investigate the interaction between E.coli and Streptomyces coelicolor A3 (2) for the increased production of undecylprodigiosin and identify the E. coli actives mediating this inter-species interaction. The antibiotics of interest were the red-pigmented undecylprodigiosin and blue-pigmented actinorhodin. Pure cultures of S. coelicolor in a defined medium produced higher concentrations of actinorhodin compared to those of undecylprodigiosin. The latter however, is more important due to its immunosuppressive and antitumor properties. As a strategy to increase undecylprodigiosin production, we added separately, live cells and heat-killed cells of E. coli C600, and the cell-free supernatant of E. coli culture to S. coelicolor cultures in shake flasks. The interaction with live cells of E. coli altered the antibiotic production pattern and undecylprodigiosin production was enhanced by 3.5-fold compared to the pure cultures of S. coelicolor and actinorhodin decreased by 15-fold. The heat-killed cells of E. coli however, had no effect on antibiotic production. In all cases, growth and glucose consumption of S. coelicolor remained almost the same as those observed in the pure culture indicating that the changes in antibiotic production were not due to nutritional stress. Results with cell-free supernatant of E. coli culture indicated that the interaction between S. coelicolor and E. coli was mediated via diffusible molecule(s). Using a set of extraction procedures and agar-well diffusion bioassays, we isolated and preliminarily identified a class of compounds. For the preliminary verification, we added the compound which was the common chemical structural moiety in this class of compounds to the pure S. coelicolor cultures. We observed similar effects on antibiotic production as with the live E. coli cells and their supernatant indicating that this class of compounds secreted by E. coli indeed could act as actives during interspecies
El-Naggar, Noura El-Ahmady; Deraz, Sahar F.; Soliman, Hoda M.; El-Deeb, Nehal M.; El-Ewasy, Sara M.
L-asparaginase is an important enzyme as therapeutic agents used in combination with other drugs in the treatment of acute lymphoblastic leukemia. A newly isolated actinomycetes strain, Streptomyces sp. NEAE-82, was potentially producing extracellular L-asparaginase, it was identified as Streptomyces fradiae NEAE-82, sequencing product was deposited in the GenBank database under accession number KJ467538. L-asparaginase was purified from the crude enzyme using ammonium sulfate precipitation, dialysis and ion exchange chromatography using DEAE Sepharose CL-6B. Further the kinetic studies of purified enzyme were carried out. The optimum pH, temperature and incubation time for maximum L-asparaginase activity were found to be 8.5, 40 °C and 30 min, respectively. The optimum substrate concentration was found to be 0.06 M. The Km and Vmax of the enzyme were 0.01007 M and 95.08 Uml−1min−1, respectively. The half-life time (T1/2) was 184.91 min at 50 °С, while being 179.53 min at 60 °С. The molecular weight of the subunits of L-asparaginase was found to be approximately 53 kDa by SDS–PAGE analysis. The purified L-asparaginase showed a final specific activity of 30.636 U/mg protein and was purified 3.338-fold. The present work for the first time reported more information in the production, purification and characterization of L-asparaginase produced by newly isolated actinomycetes Streptomyces fradiae NEAE-82. PMID:27605431
Ilic-Tomic, Tatjana; Genčić, Marija S; Živković, Milena Z; Vasiljevic, Branka; Djokic, Lidija; Nikodinovic-Runic, Jasmina; Radulović, Niko S
Herein, a novel soil bacterium Streptomyces sp. NP10 able to grow outside usual streptomycetes optimum conditions (e.g., at 4 °C, pH 9 and high NaCl concentration), exhibiting atypical hemolytic, DNAse, and cellulolytic activities, is described. This strain produces and excretes into the growth medium large amounts of free long-chain fatty acids (FAs). A concurrent lipidomics study revealed a large structural diversity of FAs with over 50 different n- and branched-chain, (un)saturated, and cyclopropane FAs (C7-C30) produced by this strain. Two of these, i-17:0cy9-10 and a-18:0cy9-10, represent new natural products and the first ever identified branched cyclopropane FAs. Both free and bound lipid profiles of Streptomyces sp. NP10 were dominated by saturated branched chain FAs (i-14:0, a-15:0, and i-16:0). Although these free FAs showed only a moderate antimicrobial activity, our results suggest that they could have an ecophysiological role in interspecies signaling with another soil microorganism Pseudomonas aeruginosa. This work represents the first comprehensive report on the structural diversity and complexity of the free FA pool in Streptomyces. A naturally occurring streptomycete, such as Streptomyces sp. NP10, which secretes significant amounts of free long-chain FAs (non-cytotoxic) into the medium, could be useful in microbial biodiesel production.
The ARS Culture Collection (NRRL) currently contains 7569 strains within the family Streptomycetaceae but 4368 of them have not been characterized to the species level. A gene sequence database using the Bacterial Isolate Genomic Sequence Database package (BIGSdb) (Jolley & Maiden, 2010) is availabl...
Imai, Shunsuke; Ichikawa, Kazuya; Muramatsu, Yuki; Kasai, Daisuke; Masai, Eiji; Fukuda, Masao
Rubber-degrading bacteria were screened for the production of clearing zones around their colonies on latex overlay agar plates. Novel three bacteria, Streptomyces sp. strain LCIC4, Actinoplanes sp. strain OR16, and Methylibium sp. strain NS21, were isolated. To the best of our knowledge, this is the first report on the isolation of a Gram-negative rubber-degrading bacterium other than γ-proteobacteria. Gel permeation chromatography analysis revealed that these strains degraded poly(cis-1,4-isoprene) to low-molecular-weight products. The occurrence of aldehyde groups in the degradation products by NS21 was suggested by staining with Schiff's reagent and ¹H-nuclear magnetic resonance spectroscopy. The lcp gene of LCIC4, which showed 99% amino acid sequence identity with that of Streptomyces sp. strain K30, was cloned, and contained a putative twin-arginine motif at its N terminus. It is located next to oxiB, which is estimated to be responsible for oxidation of degradation intermediate of rubber in K30. Southern hybridization analysis using LCIC4 lcp probe revealed the presence of a lcp-homolog in OR16. These results suggest that the lcp-homologs are involved in rubber degradation in LCIC4 and OR16.
Lin, Lan; Ge, Hui Ming; Yan, Tong; Qin, Yan Hua; Tan, Ren Xiang
Each plant species in nature harbors endophytes, a community of microbes living within host plants without causing any disease symptom. However, the exploitation of endophyte-based phytoprotectants is hampered by the paucity of mechanistic understandings of endophyte-plant interaction. We here reported two endophytic Streptomyces isolates IFB-A02 and IFB-A03 recovered from a stress-tolerant dicotyledonous plant Artemisia annua L. After the determination of their non-pathogenicity at the genomic level and from the toxin (thaxtomin A, TXT) level, the endophytism of both isolates was supported by their successful colonization in planta. Of the two endophytes, IFB-A03 was further studied for the mechanism of endophyte-conferred phytoprotection owing to its plant growth promotion in model eudicot Arabidopsis thaliana. Using the endophyte-Arabidopsis co-cultivation system into which pathogenic Streptomyces scabies was introduced, we demonstrated that IFB-A03 pre-inoculation could activate the salicylic acid (SA)-mediated plant defense responses upon pathogen challenge. Moreover, IFB-A03 was shown to partially rescue the defense deficiency in eds5 (enhanced disease susceptibility 5) Arabidopsis mutants, putatively acting at the upstream of SA accumulation in the defense signaling pathway associated with the systemic acquired resistance (SAR). These data suggest that endophytic Streptomyces sp. IFB-A03 could be a promising candidate for biocontrol agents against S. scabies--a causative pathogen of common scab diseases prevailing in agronomic systems.
Hirano, Satomi; Sekine, Kazuhisa; Handa, Tetsurou; Nakano, Minoru
Phospholipase D (PLD)-mediated hydrolysis of phosphatidylcholine (PC) in large unilamellar vesicles (LUVs) consisting of PC and either glycerol monooleate (GMO) or methyl oleate (MeO) were monitored in situ and in real time by using a choline oxidase-immobilized oxygen electrode. This technique revealed reaction differences between 2 bacterial PLDs. PLD from Streptomyces chromofuscus, which is closely homologous to bacterial alkaline phosphatase, hydrolyzed only 6% of surface PC owing to product inhibition. The catalytic activity of this enzyme was not sensitive to the addition of GMO. On the other hand, typical bacterial PLD from Streptomyces sp. was found to hydrolyze all the PC molecules at the outer surface of LUVs suggesting that this enzyme is free from product inhibition. Introduction of GMO or MeO into the bilayer increased exposure of the PC headgroup and facilitated PC hydrolysis mediated by PLD from Streptomyces sp. GMO and MeO have the same lipophilic tail but the latter lacks hydroxyl groups on its polar head. From kinetic analysis by using the Michaelis-Menten model extended to the reaction at the interface, these compounds were found to activate PLD from Streptomyces sp. in different ways, i.e., MeO increased the protein binding to membranes and GMO stimulated the enzyme-substrate complex formation at membrane surface. Copyright © 2012 Elsevier B.V. All rights reserved.
Characterization of a silent sesquiterpenoid biosynthetic pathway in Streptomyces avermitilis controlling epi-isozizaene albaflavenone biosynthesis and isolation of a new oxidized epi-isozizaene metabolite.
Takamatsu, Satoshi; Lin, Xin; Nara, Ayako; Komatsu, Mamoru; Cane, David E; Ikeda, Haruo
The genome-sequenced, Gram-positive bacterium Streptomyces avermitilis harbours an orthologue (SAV_3032) of the previously identified epi-isozizaene synthase (SCO5222) in Streptomyces coelicolor A3(2). The sav3032 is translationally coupled with the downstream sav3031 gene encoding the cytochrome P450 CYP170A2 analogous to SCO5223 (CYP170A1) of S. coelicolor A3(2), which exhibits a similar translation coupling. Streptomyces avermitilis did not produce epi-isozizaene or any of its oxidized derivatives, albaflavenols and albaflavenone, under in any culture conditions examined. Nonetheless, recombinant SAV_3032 protein expressed in Escherichia coli catalysed the Mg²+-dependent cyclization of farnesyl diphosphate to epi-isozizaene. To effect the production of epi-isozizaene in S. avermitilis, the sav3032 gene was cloned and placed under control of a copy of the native S. avermitilis promoter rpsJp (sav4925). The derived expression construct was introduced by transformation into a large-deletion mutant of S. avermitilis SUKA16 and the resulting transformants accumulated epi-isozizaene. The previously characterized oxidized epi-isozizaene metabolites (4R)- and (4S)-albaflavenols and albaflavenone, as well as a previously undescribed doubly oxidized epi-isozizaene derivative were isolated from cultures of S. avermitilis SUKA16 transformants in which sav3032 was coexpressed with the P450-encoding sav3031. This new metabolite was identified as 4β,5β-epoxy-2-epi-zizaan-6β-ol which is most likely formed by oxidation of (4S)-albaflavenol.
El-Naggar, Noura El-Ahmady; El-Ewasy, Sara M.
In this present study, a newly isolated strain, Streptomyces sp. NEAE-H, capable of producing high amount of black extracellular melanin pigment on peptone-yeast extract iron agar and identified as Streptomyces glaucescens NEAE-H. Plackett–Burman statistical design was conducted for initial screening of 17 independent (assigned) variables for their significances on melanin pigment production by Streptomyces glaucescens NEAE-H. The most significant factors affecting melanin production are incubation period, protease-peptone and ferric ammonium citrate. The levels of these significant variables and their interaction effects were optimized by using face-centered central composite design. The maximum melanin production (31.650 μg/0.1 ml) and tyrosinase activity (6089.10 U/ml) were achieved in the central point runs under the conditions of incubation period (6 days), protease-peptone (5 g/L) and ferric ammonium citrate (0.5 g/L). Melanin pigment was recovered by acid-treatment. Higher absorption of the purified melanin pigment was observed in the UV region at 250 nm. It appeared to have defined small spheres by scanning electron microscopy imaging. The maximum melanin yield was 350 mg dry wt/L of production medium. In vitro anticancer activity of melanin pigment was assayed against skin cancer cell line using MTT assay. The IC50 value was 16.34 ± 1.31 μg/ml for melanin and 8.8 ± 0.5 μg/ml for standard 5-fluorouracil. PMID:28195138
Cheryl P. Andam
Full Text Available We show that Streptomyces biogeography in soils across North America is influenced by the regional diversification of microorganisms due to dispersal limitation and genetic drift. Streptomyces spp. form desiccation-resistant spores, which can be dispersed on the wind, allowing for a strong test of whether dispersal limitation governs patterns of terrestrial microbial diversity. We employed an approach that has high sensitivity for determining the effects of genetic drift. Specifically, we examined the genetic diversity and phylogeography of physiologically similar Streptomyces strains isolated from geographically distributed yet ecologically similar habitats. We found that Streptomyces beta diversity scales with geographic distance and both beta diversity and phylogenetic diversity manifest in a latitudinal diversity gradient. This pattern of Streptomyces biogeography resembles patterns seen for diverse species of plants and animals, and we therefore evaluated these data in the context of ecological and evolutionary hypotheses proposed to explain latitudinal diversity gradients. The data are consistent with the hypothesis that niche conservatism limits dispersal, and historical patterns of glaciation have limited the time for speciation in higher-latitude sites. Most notably, higher-latitude sites have lower phylogenetic diversity, higher phylogenetic clustering, and evidence of range expansion from lower latitudes. In addition, patterns of beta diversity partition with respect to the glacial history of sites. Hence, the data support the hypothesis that extant patterns of Streptomyces biogeography have been driven by historical patterns of glaciation and are the result of demographic range expansion, dispersal limitation, and regional diversification due to drift.
Alshaibani, Muhanna M; Jalil, Juriyati; Sidik, Nik M; Edrada-Ebel, Ruangelie; Zin, Noraziah M
Background Zingiber spectabile, commonly known as Beehive Ginger, is used as an ethnobotanical plant in many countries as an appetizer or to treat stomachache, toothache, muscle sprain, and as a cure for swelling, sores and cuts. This is the first report of isolation of Streptomyces strain from the root of this plant. Strain Universiti Kebangsaan 25 (SUK 25) has a very high activity to produce secondary metabolites against methicillin-resistant Staphylococcus aureus (MRSA), which is associated with high morbidity and mortality rates due to acquired multidrug resistance genes and causes medication failure in some clinical cases worldwide. Phylogenetic analysis based on the 16S ribosomal RNA gene sequence exhibited that the most closely related strain was Streptomyces omiyaensis NBRC 13449T (99.0% similarity). Aim This study was conducted to carry out the extraction, identification, and biological evaluation of active metabolites isolated from SUK 25 against three MRSA strains, namely, MRSA ATCC 43300, MRSA ATCC 33591, and MRSA ATCC 49476. Materials and methods The production of secondary metabolites by this strain was optimized through Thronton’s media. Isolation, purification, and identification of the bioactive compounds were carried out using reversed-phase high-performance liquid chromatography, high-resolution mass spectrometry, Fourier transform infrared, and one-dimensional and two-dimensional nuclear magnetic resonance. Results During screening procedure, SUK 25 exhibited good antimicrobial potential against several strains of MRSA. The best biological activity was shown from fraction number VII and its subfractions F2 and F3 with minimum inhibitory concentration values at 16 µg/mL and 8 µg/mL, respectively. These two subfractions were identified as diketopiperazine cyclo-(tryptophanyl-prolyl) and chloramphenicol. Conclusion On the basis of obtained results, SUK 25 isolated from Z. spectabile can be regarded as a new valuable source to produce secondary
Hulcr, Jiri; Adams, Aaron S; Raffa, Kenneth; Hofstetter, Richard W; Klepzig, Kier D; Currie, Cameron R
Recent studies have revealed several examples of intimate associations between insects and Actinobacteria, including the Southern Pine Beetle Dendroctonus frontalis and the Spruce Beetle Dendroctonus rufipennis. Here, we surveyed Streptomyces Actinobacteria co-occurring with 10 species of Dendroctonus bark beetles across the United States, using both phylogenetic and community ecology approaches. From these 10 species, and 19 other scolytine beetles that occur in the same trees, we obtained 154 Streptomyces-like isolates and generated 16S sequences from 134 of those. Confirmed 16S sequences of Streptomyces were binned into 36 distinct strains using a threshold of 0.2% sequence divergence. The 16S rDNA phylogeny of all isolates does not correlate with the distribution of strains among beetle species, localities, or parts of the beetles or their galleries. However, we identified three Streptomyces strains occurring repeatedly on Dendroctonus beetles and in their galleries. Identity of these isolates was corroborated using a house-keeping gene sequence (efTu). These strains are not confined to a certain species of beetle, locality, or part of the beetle or their galleries. However, their role as residents in the woodboring insect niche is supported by the repeated association of their 16S and efTu from across the continent, and also having been reported in studies of other subcortical insects.
Hoshino, Shotaro; Okada, Masahiro; Wakimoto, Toshiyuki; Zhang, Huiping; Hayashi, Fumiaki; Onaka, Hiroyasu; Abe, Ikuro
A terrestrial bacterium, Streptomyces sp. NZ-6, produced niizalactams A-C (1-3), unprecedented di- and tricyclic macrolactams, by coculturing with the mycolic acid-containing bacterium Tsukamurella pulmonis TP-B0596. Their complete structures, including absolute configurations, were elucidated on the basis of spectroscopic data and chemical derivatization. Their unique skeletons are proposed to be biosynthesized from a common 26-membered macrolactam intermediate by SN2 cyclization or an intramolecular Diels-Alder reaction.
Johansson, Elisabet; Reponen, Tiina; Meller, Jarek; Vesper, Stephen; Yadav, Jagjit
Both Streptomyces species and mold species have previously been isolated from moisture-damaged building materials; however, an association between these two groups of microorganisms in indoor environments is not clear. In this study, we used a culture-independent method, PCR-denaturing gradient gel electrophoresis (PCR-DGGE), to investigate the composition of the Streptomyces community in house dust. Twenty-three dust samples each from two sets of homes categorized as high-mold and low-mold based on mold-specific quantitative PCR analysis were used in the study. Taxonomic identification of prominent bands was performed by cloning and sequencing. Associations between DGGE amplicon band intensities and home mold status were assessed using univariate analyses as well as multivariate recursive partitioning (decision trees) to test the predictive value of combinations of bands intensities. In the final classification tree, a combination of two bands was significantly associated with mold status of the home (p = 0.001). The sequence corresponding to one of the bands in the final decision tree matched a group of Streptomyces species that included Streptomyces coelicolor and Streptomyces sampsonii, both of which have been isolated from moisture-damaged buildings previously. The closest match for the majority of sequences corresponding to a second band consisted of a group of Streptomyces species that included Streptomyces hygroscopicus, an important producer of antibiotics and immunosuppressors. Taken together, the study showed that DGGE can be a useful tool for identifying bacterial species that may be more prevalent in mold-damaged buildings.
Breinholt, J.; Gürtler, Hanne; Kjær, Anders
A colourless, crystalline metabolite, C14H12ClNO4, named streptopyrrole, has been isolated from submerged fermentation cultures of Streptomyces armeniacus by extraction, followed by chromatographic purification. Its tricyclic molecular framework, seemingly without natural product precedents...
Rajaretinam Rajesh Kannan; Appadurai Muthamil Iniyan; Vincent Samuel Gnana Prakash
Objective: The aim of the present study was to isolate the anti-MRSA (Methicillin ResistantStaphylococcus aureus ) molecule from the Mangrove symbiont Streptomyces and its biomedical studies in Zebrafish embryos. Methods: MRSA was isolated from the pus samples of Colachal hospitals and confirmed by amplification of mecA gene. Anti-MRSA molecule producing strain was identified by 16s rRNA gene sequencing. Anti-MRSA compound production was optimized by Solid State Fermentation (SSF) and the purification of the active molecule was carried out by TLC and RP-HPLC. The inhibitory concentration and LC50 were calculated using Statistical software SPSS. The Biomedical studies including the cardiac assay and organ toxicity assessment were carried out in Zebrafish. Results: The bioactive anti-MRSA small molecule A2 was purified by TLC with Rf value of 0.37 with 1.389 retention time at RP-HPLC. The Inhibitory Concentration of the purified molecule A2 was 30 μg/mL but, the inhibitory concentration of the MRSA in the infected embryo was 32-34 μg/mL for TLC purified molecule A2 with LC50 mean value was 61.504 μg/mL. Zebrafish toxicity was assessed in 48-60 μg/mL by observing the physiological deformities and the heart beat rates (HBR) of embryos for anti MRSA molecule showed the mean of 41.33-41.67 HBR/15 seconds for 40 μg/mL and control was 42.33-42.67 for 15 seconds which significantly showed that the anti-MRSA molecule A2 did not affected the HBR. Conclusions:Anti-MRSA molecule from Streptomyces sp PVRK-1 was isolated and biomedical studies in Zebrafish model assessed that the molecule was non toxic at the minimal inhibitory concentration of MRSA.
Jog, Rahul; Pandya, Maharshi; Nareshkumar, G; Rajkumar, Shalini
The application of plant-growth-promoting rhizobacteria (PGPR) at field scale has been hindered by an inadequate understanding of the mechanisms that enhance plant growth, rhizosphere incompetence and the inability of bacterial strains to thrive in different soil types and environmental conditions. Actinobacteria with their sporulation, nutrient cycling, root colonization, bio-control and other plant-growth-promoting activities could be potential field bio-inoculants. We report the isolation of five rhizospheric and two root endophytic actinobacteria from Triticum aestivum (wheat) plants. The cultures exhibited plant-growth-promoting activities, namely phosphate solubilization (1916 mg l(-1)), phytase (0.68 U ml(-1)), chitinase (6.2 U ml(-1)), indole-3-acetic acid (136.5 mg l(-1)) and siderophore (47.4 mg l(-1)) production, as well as utilizing all the rhizospheric sugars under test. Malate (50-55 mmol l(-1)) was estimated in the culture supernatant of the highest phosphate solublizer, Streptomyces mhcr0816. The mechanism of malate overproduction was studied by gene expression and assays of key glyoxalate cycle enzymes - isocitrate dehydrogenase (IDH), isocitrate lyase (ICL) and malate synthase (MS). The significant increase in gene expression (ICL fourfold, MS sixfold) and enzyme activity (ICL fourfold, MS tenfold) of ICL and MS during stationary phase resulted in malate production as indicated by lowered pH (2.9) and HPLC analysis (retention time 13.1 min). Similarly, the secondary metabolites for chitinase-independent biocontrol activity of Streptomyces mhcr0817, as identified by GC-MS and (1)H-NMR spectra, were isoforms of pyrrole derivatives. The inoculation of actinobacterial isolate mhce0811 in T. aestivum (wheat) significantly improved plant growth, biomass (33%) and mineral (Fe, Mn, P) content in non-axenic conditions. Thus the actinobacterial isolates reported here were efficient PGPR possessing significant antifungal activity and may have potential field
Hsiao, Nai-Hua; Kirby, Ralph
DNA/DNA microarray hybridization was used to compare the genome content of Streptomyces avermitilis, Streptomyces cattleya, Streptomyces maritimus and Kitasatospora aureofaciens with that of Streptomyces coelicolor A3(2). The array data showed an about 93% agreement with the genome sequence data available for S. avermitilis and also showed a number of trends in the genome structure for Streptomyces and closely related Kitasatospora. A core central region was well conserved, which might be predicted from previous research and this was linked to a low degree of gene conservation in the terminal regions of the linear chromosome across all four species. Between these regions there are two areas of intermediate gene conservation by microarray analysis where gene synteny is still detectable in S. avermitilis. Nonetheless, a range of conserved genes could be identified within the terminal regions. Variation in the genes involved in differentiation, transcription, DNA replication, etc. provides interesting insights into which genes in these categories are generally conserved and which are not. The results also provide target priorities for possible gene knockouts in a group of bacteria with a very large numbers of genes with unknown functions compared to most bacterial species.
Ryan F Seipke
Full Text Available Attine ants are dependent on a cultivated fungus for food and use antibiotics produced by symbiotic Actinobacteria as weedkillers in their fungus gardens. Actinobacterial species belonging to the genera Pseudonocardia, Streptomyces and Amycolatopsis have been isolated from attine ant nests and shown to confer protection against a range of microfungal weeds. In previous work on the higher attine Acromyrmex octospinosus we isolated a Streptomyces strain that produces candicidin, consistent with another report that attine ants use Streptomyces-produced candicidin in their fungiculture. Here we report the genome analysis of this Streptomyces strain and identify multiple antibiotic biosynthetic pathways. We demonstrate, using gene disruptions and mass spectrometry, that this single strain has the capacity to make candicidin and multiple antimycin compounds. Although antimycins have been known for >60 years we report the sequence of the biosynthetic gene cluster for the first time. Crucially, disrupting the candicidin and antimycin gene clusters in the same strain had no effect on bioactivity against a co-evolved nest pathogen called Escovopsis that has been identified in ∼30% of attine ant nests. Since the Streptomyces strain has strong bioactivity against Escovopsis we conclude that it must make additional antifungal(s to inhibit Escovopsis. However, candicidin and antimycins likely offer protection against other microfungal weeds that infect the attine fungal gardens. Thus, we propose that the selection of this biosynthetically prolific strain from the natural environment provides A. octospinosus with broad spectrum activity against Escovopsis and other microfungal weeds.
Seipke, Ryan F; Barke, Jörg; Brearley, Charles; Hill, Lionel; Yu, Douglas W; Goss, Rebecca J M; Hutchings, Matthew I
Attine ants are dependent on a cultivated fungus for food and use antibiotics produced by symbiotic Actinobacteria as weedkillers in their fungus gardens. Actinobacterial species belonging to the genera Pseudonocardia, Streptomyces and Amycolatopsis have been isolated from attine ant nests and shown to confer protection against a range of microfungal weeds. In previous work on the higher attine Acromyrmex octospinosus we isolated a Streptomyces strain that produces candicidin, consistent with another report that attine ants use Streptomyces-produced candicidin in their fungiculture. Here we report the genome analysis of this Streptomyces strain and identify multiple antibiotic biosynthetic pathways. We demonstrate, using gene disruptions and mass spectrometry, that this single strain has the capacity to make candicidin and multiple antimycin compounds. Although antimycins have been known for >60 years we report the sequence of the biosynthetic gene cluster for the first time. Crucially, disrupting the candicidin and antimycin gene clusters in the same strain had no effect on bioactivity against a co-evolved nest pathogen called Escovopsis that has been identified in ∼30% of attine ant nests. Since the Streptomyces strain has strong bioactivity against Escovopsis we conclude that it must make additional antifungal(s) to inhibit Escovopsis. However, candicidin and antimycins likely offer protection against other microfungal weeds that infect the attine fungal gardens. Thus, we propose that the selection of this biosynthetically prolific strain from the natural environment provides A. octospinosus with broad spectrum activity against Escovopsis and other microfungal weeds.
Baba, Mohd Shukri; Zin, Noraziah Mohamad; Hassan, Zainal Abidin Abu; Latip, Jalifah; Pethick, Florence; Hunter, Iain S; Edrada-Ebel, RuAngelie; Herron, Paul R
Endophytic bacteria, such as Streptomyces, have the potential to act as a source for novel bioactive molecules with medicinal properties. The present study was aimed at assessing the antimalarial activity of crude extract isolated from various strains of actinobacteria living endophytically in some Malaysian medicinal plants. Using the four day suppression test method on male ICR strain mice, compounds produced from three strains of Streptomyces (SUK8, SUK10, and SUK27) were tested in vivo against Plasmodium berghei PZZ1/100 in an antimalarial screen using crude extracts at four different concentrations. One of these extracts, isolated from Streptomyces SUK10 obtained from the bark of Shorea ovalis tree, showed inhibition of the test organism and was further tested against P. berghei-infected mice for antimalarial activity at different concentrations. There was a positive relationship between the survival of the infected mouse group treated with 50 µg/kg body weight (bw) of ethyl acetate-SUK10 crude extract and the ability to inhibit the parasites growth. The parasite inhibition percentage for this group showed that 50% of the mice survived for more than 90 days after infection with the parasite. The nucleotide sequence and phylogenetic tree suggested that Streptomyces SUK10 may constitute a new species within the Streptomyces genus. As part of the drug discovery process, these promising finding may contribute to the medicinal and pharmaceutical field for malarial treatment.
Duraipandiyan, V; Al-Dhabi, N A; Balachandran, C; Raj, M Karunai; Arasu, M Valan; Ignacimuthu, S
Streptomyces sp. isolate ERI-26 was obtained from the Nilgiris forest soil of Western Ghats, Tamil Nadu, India. Novel anthraquinone compound was isolated from the active fraction 5; it was identified by spectroscopical data using UV, IR, NMR and MASS. The isolated compound 1,5,7-trihydroxy-3-hydroxy methyl anthraquinone was tested against bacteria and fungi at minimum inhibitory concentration level. The compound showed significant antimicrobial activity against bacteria, Staphylococcus aureus at 125 μg/ml, Staphylococcus epidermidis at 62.5 μg/m, Bacillus subtilis at 31.25 μg/ml, fungi; Epidermophyton floccosum at 62.5 μg/ml, Aspergillus niger at 31.25 μg/ml, Aspergiller flavus at 31.25 μg/ml, Trichophyton rubrum at 62.5 μg/ml and Botrytis cinerea at 62.5 μg/ml. The isolated compound was subjected to molecular docking studies for the inhibition of TtgR, topoisomerase IV and AmpC β-lactamase enzymes which are targets for antimicrobials. Docking studies of the compound showed low docking energy indicating its usefulness as antimicrobial agent. 1,5,7-Trihydroxy-3-hydroxy methyl anthraquinone is new, and its antimicrobial and molecular docking properties are reported for the first time.
The present work studies the chemical constituents from marine-derived streptomyces 3320# and their antitumor activities. The n-BuOH extract of the ferment broth of 3320# was chromatographed on silica gel, Sephadex LH-20, ODS columns and HPLC to separate the compounds with antitoumor activities. Their structures were identified using IR, UV, NMR, MS spectroscopic techniques and compared with published data. The antitumor activities of the isolates were assayed using SRB method and flow cytometry assay, accompanied with the morphological observation of the cells under light microscope against mammalian tsFT210 cells. Ten compounds, cyclo-(Ala-Leu) 1, cyclo-(Ala-Ile) 2, cyclo-(Ala-Val) 3, cyclo-(Phe- Pro) 4, cyclo-(Phe-Gly) 5, cyclo-(Leu-Pro) 6, 1-methyl-1, 2, 3, 4-tetrahydro-β-carboline-3-carboxylic acid 7, N-(4-hydroxyphenethyl) acetamide 8, 4-methyoxy-1-(2-hydroxy) ethylbenzene 9 and uridine 10, were isolated from the ferment broth of streptomyces 3320#. Among them, compounds 6, 7, 8 and 10 showed potent cytotoxicity against the tsFT210 cell with the IC50 values of 3 . 6, 7 . 2, 5 . 2 and 1 . 6 mmol L - 1, respectively. Compounds 8, 10 also exhibited apoptosis inducing activity under 2 . 0 mmol L - 1. Compounds 6, 7, 8 and 10 are the principle bioactive constituents responsible for the antitumor activities of marine streptomyces 3320# . Compound 7 was isolated from this species for the first time.
Consistent, effective biological control of soilborne pathogens has been difficult to achieve in the field. Most research regarding biological control efficacy has focused on the biological control agent itself (i.e. dose, formulation, survival) or on its interaction with the rhizosphere and edaphi...
Disease control of soilborne pathogens by biological control agents has often been inconsistent under field conditions. One factor that may contribute to this inconsistency is the variability in response among pathogen populations and/or communities to the selected biological control agent. One hund...
Full Text Available Muhanna M Alshaibani,1 Juriyati Jalil,2 Nik M Sidik,3 Ruangelie Edrada-Ebel,4 Noraziah M Zin1 1Programme of Biomedical Science, School of Diagnostic and Applied Health Sciences, Faculty of Health Sciences, 2Drug and Herbal Research Centre, Faculty of Pharmacy, Universiti Kebangsaan Malaysia, Kuala Lumpur, 3School of Bioscience and Biotechnology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, Bangi, Malaysia; 4Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, Glasgow, Scotland Background: Zingiber spectabile, commonly known as Beehive Ginger, is used as an ethnobotanical plant in many countries as an appetizer or to treat stomachache, toothache, muscle sprain, and as a cure for swelling, sores and cuts. This is the first report of isolation of Streptomyces strain from the root of this plant. Strain Universiti Kebangsaan 25 (SUK 25 has a very high activity to produce secondary metabolites against methicillin-resistant Staphylococcus aureus (MRSA, which is associated with high morbidity and mortality rates due to acquired multidrug resistance genes and causes medication failure in some clinical cases worldwide. Phylogenetic analysis based on the 16S ribosomal RNA gene sequence exhibited that the most closely related strain was Streptomyces omiyaensis NBRC 13449T (99.0% similarity. Aim: This study was conducted to carry out the extraction, identification, and biological evaluation of active metabolites isolated from SUK 25 against three MRSA strains, namely, MRSA ATCC 43300, MRSA ATCC 33591, and MRSA ATCC 49476. Materials and methods: The production of secondary metabolites by this strain was optimized through Thronton’s media. Isolation, purification, and identification of the bioactive compounds were carried out using reversed-phase high-performance liquid chromatography, high-resolution mass spectrometry, Fourier transform infrared, and one-dimensional and two-dimensional nuclear magnetic
Wellington, E M; Stackebrandt, E; Sanders, D; Wolstrup, J; Jorgensen, N O
Species classified within the genus Kitasatosporia share many of the phenotypic characteristics typical of streptomycetes. By using a probabilistic identification scheme, they were identified with Streptomyces exfoliatus cluster 5, a species group within Streptomyces. The four species studied hybridized with a 16S rRNA genus probe for Streptomyces spp., indicating a close relationship between the two genera. The kitasatosporias were resistant to selected polyvalent streptomycete phages tested. Quantitative analysis showed that meso-diaminopimelic acid varied from 49 to 89% in Kitasatosporia species and from 1 to 16% in Streptomyces species depending on growth conditions. On the basis of 16S rRNA analysis, it is proposed to reduce Kitasatosporia to synonymy with Streptomyces. As a result, the new names proposed are Streptomyces mediocidicus comb. nov., Streptomyces phosalacineus comb. nov., Streptomyces setae comb. nov., and Streptomyces griseolosporeus comb. nov., nom. nov.
江冰娅; 赵薇; 李书芬; 刘红宇; 余利岩; 武临专
目的：对醌霉素产生菌——Streptomyces sp. CPCC 200497产生的次级代谢产物进行深入研究。方法将 Streptomyces sp. CPCC 200497进行固体发酵，用高效硅胶板薄层色谱、中低压制备色谱和高效液相色谱等方法对发酵培养物的乙酸乙酯提取物进行分析和纯化；通过质谱和 NMR 等对获得的单体化合物进行结构鉴定。结果从 Streptomyces sp. CPCC 200497发酵物的乙酸乙酯提取物中发现了两个吡咯里西啶类生物碱，分别为bohemamine 和 bohemamine B。结论从醌霉素产生菌 Streptomyces sp. CPCC 200497中首次分离得到 bohemamine 类化合物，并通过1D 和2D-NMR 对化合物的核磁数据进行了准确归属，为国内首次对该类化合物的分离鉴定进行报道。%Objective To study in depth the secondary metabolites from Streptomyces sp. CPCC 200497, a quinomycins producer. Methods Silica gel TLC, medium-low-pressure preparative chromatography and reversed phase HPLC were used to analyze the EtOAc extract of agar culture of Streptomyces sp. CPCC 200497 or purify compounds of interests. LC-MS and NMR were employed to identify the target compounds. Results Bohemamine and bohemamine B, two pyrrolizidine alkaloids were found in ethyl acetate extracts of fermentation from Streptomyces sp. CPCC 200497 and characterized for their structures. Conclusion Bohemamines are identified for the first time as secondary metabolites from Streptomyces sp. CPCC 200497, a quinomycins producing strain. Their structures are established on the basis of extensive spectroscopic data. This is the first report of bohemamines in China.
Full Text Available Streptomyces species are prolific producers of bioactive metabolites, such as β-lactone-containing lipstatin produced by Streptomyces toxytricini, an intermediate used in semi-synthetic process for production of anti-obesity drug orlistat. Understanding the distribution of identical or structurally similar molecules produced by a taxonomic group is of particular importance when trying to isolate novel biologically active compounds or strains producing known metabolites of medical importance with potentially improved properties. Until now, only two independent isolates of S. toxytricini species have been known to be producers of lipstatin. According to the current taxonomic criteria, S. toxytricini belongs to Streptomyces lavendulae phenotypic cluster. Taxonomy-based dereplication approach coupled with in vitro assay was applied to screen the S. lavendulae phenotypic cluster for production of lipstatin-like lipase inhibitors using synthetic p-nitrophenol derivatives of C4 and C16 lipids. Screening the available strains from public collections belonging to S. lavendulae phenotypic cluster, high lipase inhibitory activity was identified in the Streptomyces virginiae CBS 314.55 culture supernatants. HPLC and LC-MS/MS confirmed lipstatin production by a new Streptomyces species for the first time. We have demonstrated that the new lipstatin-producing strain S. virginiae morphologically and physiologically differs from S. toxytricini substantially; however, the production capacity of the newly identified lipstatin-producing species S. virginiae is comparable to S. toxytricini. We have thus demonstrated the effectiveness of a simple and affordable dereplication approach for identification of potentially novel and useful industrial strains available in public culture collections.
Full Text Available Metabolomics and genomics are two complementary platforms for analyzing an organism as they provide information on the phenotype and genotype, respectively. These two techniques were applied in the dereplication and identification of bioactive compounds from a Streptomyces sp. (SM8 isolated from the sponge Haliclona simulans from Irish waters. Streptomyces strain SM8 extracts showed antibacterial and antifungal activity. NMR analysis of the active fractions proved that hydroxylated saturated fatty acids were the major components present in the antibacterial fractions. Antimycin compounds were initially putatively identified in the antifungal fractions using LC-Orbitrap. Their presence was later confirmed by comparison to a standard. Genomic analysis of Streptomyces sp. SM8 revealed the presence of multiple secondary metabolism gene clusters, including a gene cluster for the biosynthesis of the antifungal antimycin family of compounds. The antimycin gene cluster of Streptomyces sp. SM8 was inactivated by disruption of the antimycin biosynthesis gene antC. Extracts from this mutant strain showed loss of antimycin production and significantly less antifungal activity than the wild-type strain. Three butenolides, 4,10-dihydroxy-10-methyl-dodec-2-en-1,4-olide (1, 4,11-dihydroxy-10-methyl-dodec-2-en-1,4-olide (2, and 4-hydroxy-10-methyl-11-oxo-dodec-2-en-1,4-olide (3 that had previously been reported from marine Streptomyces species were also isolated from SM8. Comparison of the extracts of Streptomyces strain SM8 and its host sponge, H. simulans, using LC-Orbitrap revealed the presence of metabolites common to both extracts, providing direct evidence linking sponge metabolites to a specific microbial symbiont.
Lee, Min Hwa; Cheon, Doo-Sung; Choi, Sunkeum; Lee, Bog-Hieu; Jung, Ji-Youn; Choi, Changsun
This study was conducted to determine the prevalence of Arcobacter species identified or isolated from retail meats in Korea. Multiplex PCR assays for the detection of Arcobacter species were performed for 360 chicken, 100 pork, and 106 beef samples. Arcobacter butzleri and Arcobacter cryaerophilus were detected in 18.9 and 3.3% of chicken samples, respectively. However, Arcobacter species were not found in any of the pork and beef samples. Biochemical testing of isolates selected after enrichment revealed 38 A. butzleri isolates in chicken samples, but no A. cryaerophilus isolates were detected. In this study, A. butzleri was the most prevalent Arcobacter species in chicken meat, and contamination with Arcobacter species in pork and beef may be less prevalent in Korea.
Full Text Available Streptomyces alboniger ATCC 12461 grown in brain heart infusion (BHI medium produced two extracellular serine-proteinases, denoted SP I and SP II, which were purified by ammonium sulfate precipitation and aprotinin-agarose affinity chromatography. SP I was purified 88,9-fold and SP II 66,7- fold, with 33.4% and 10.4% yield, respectively. The optimum pH for the proteinases activity, using a-N-p-tosyl-L-arginine-methyl ester (TAME as substrate, was 9-10 and the optimum temperature was 37ºC. The proteolytic activity of SP I and SP II was inhibited by aprotinin and SP I was partially inhibited by leupeptin, both serine-proteinase inhibitors. S. alboniger growth in BHI-liquid medium decreased when 5 mg/ml, 10 mg/ml of aprotinin was used, being completely inhibited with 20 mg/ml and 40 mg/ml. At the ultrastructural level, aprotinin-treated S. alboniger cells showed swelling of the bacterial body and condensation of the genetic material, probably related to the inhibition of its growth.
Nagalingam, Mohandoss; Shome, Rajeswari; Balamurugan, Vinayagamurthy; Shome, Bibek Ranjan; NarayanaRao, Krishnamsetty; Vivekananda; Isloor, Shrikrishna; Prabhudas, Krishnamsetty
Although host specificity has been observed in different species of Brucella, crossing the animal host boundary is likely to occur at any time. In this study, Bruce ladder PCR and abortus-melitensis-ovis-suis (AMOS) PCR assays were used to characterize 47 Brucella isolates from Indian origin in order to know exact species for understanding epidemiology of brucellosis. Out of them, 28, 14, and 5 isolates were found to be Brucella abortus, Brucella melitensis, and Brucella suis, respectively. Further analysis by AMOS PCR has identified that all the B. abortus isolates belong to any one of the biovar 1, 2, or 4; of the five B. suis isolates, three belong to biovar 1 and two belong to any one of the biovar 2, 3, 4, or 5. Although this multiplex Bruce ladder PCR is useful in differentiating all Brucella species, elaborate study is required to further characterize the isolates at exact biovar level.
Isolation rate and drug resistance patterns of Shigella species among ... use of water from unprotected sources and absence of latrine were the risk factors that ... to identify changes in the prevalence and antimicrobial resistance patterns of ...
... health, the continuous examination of emerging antimicrobial resistance of these ... isolated from foods and in developing effective risk management strategies. ... Keywords: Listeria species, Ready-to-eat foods, Antimicrobial susceptibility, ...
Piva, Silvia; Serraino, Andrea; Florio, Daniela; Giacometti, Federica; Pasquali, Frederique; Manfreda, Gerardo; Zanoni, Renato Giulio
This is the first report of Arcobacter spp. in rectal fecal samples from healthy water buffaloes (Bubalus bubalis) reared on a dairy farm. Arcobacter species were isolated after enrichment, and isolates were identified at species level by multiplex-polymerase chain reaction assay. Thirty samples were examined and Arcobacter spp. were isolated from 96.7% of water buffaloes tested: 38 Arcobacter spp. isolates were obtained, with A. cryaerophilus as the dominant species followed by A. butzleri and A. skirrowii. Nine animals (31%) were colonized by more than one Arcobacter species. The present study indicates that water buffaloes can harbor a variety of Arcobacter spp. and that healthy buffaloes may act as hosts. Water buffalo fecal shedding of Arcobacter spp. may be of significance to human health, considering the potential fecal contamination during harvesting of raw milk and slaughtering.
Full Text Available The genome sequence of Streptomyces coelicolor A3(2 contains more than 50 genes coding for putative lipolytic enzymes. Many studies have shown the capacity of this actinomycete to store important reserves of intracellular triacylglycerols in nutrient depletion situations. In the present study, we used genome mining of S. coelicolor to identify genes coding for putative, non-secreted esterases/lipases. Two genes were cloned and successfully overexpressed in E. coli as His-tagged fusion proteins. One of the recombinant enzymes, EstC, showed interesting cold-active esterase activity with a strong potential for the production of valuable esters. The purified enzyme displayed optimal activity at 35°C and was cold-active with retention of 25% relative activity at 10°C. Its optimal pH was 8.5-9 but the enzyme kept more than 75% of its maximal activity between pH 7.5 and 10. EstC also showed remarkable tolerance over a wide range of pH values, retaining almost full residual activity between pH 6-11. The enzyme was active toward short-chain p-nitrophenyl esters (C2-C12, displaying optimal activity with the valerate (C5 ester (k(cat/K(m = 737±77 s(-1 mM(-1. The enzyme was also very active toward short chain triglycerides such as triacetin (C2:0 and tributyrin (C4:0, in addition to showing good primary alcohol and organic solvent tolerance, suggesting it could function as an interesting candidate for organic synthesis of short-chain esters such as flavors.
Fogg, Paul C M; Haley, Joshua A; Stark, W Marshall; Smith, Margaret C M
Bacteriophages are the source of many valuable tools for molecular biology and genetic manipulation. In Streptomyces, most DNA cloning vectors are based on serine integrase site-specific DNA recombination systems derived from phage. Because of their efficiency and simplicity, serine integrases are also used for diverse synthetic biology applications. Here, we present the genome of a new Streptomyces phage, ϕJoe, and investigate the conditions for integration and excision of the ϕJoe genome. ϕJoe belongs to the largest Streptomyces phage cluster (R4-like) and encodes a serine integrase. The attB site from Streptomyces venezuelae was used efficiently by an integrating plasmid, pCMF92, constructed using the ϕJoe int-attP locus. The attB site for ϕJoe integrase was occupied in several Streptomyces genomes, including that of S. coelicolor, by a mobile element that varies in gene content and size between host species. Serine integrases require a phage-encoded recombination directionality factor (RDF) to activate the excision reaction. The ϕJoe RDF was identified, and its function was confirmed in vivo Both the integrase and RDF were active in in vitro recombination assays. The ϕJoe site-specific recombination system is likely to be an important addition to the synthetic biology and genome engineering toolbox.IMPORTANCEStreptomyces spp. are prolific producers of secondary metabolites, including many clinically useful antibiotics. Bacteriophage-derived integrases are important tools for genetic engineering, as they enable integration of heterologous DNA into the Streptomyces chromosome with ease and high efficiency. Recently, researchers have been applying phage integrases for a variety of applications in synthetic biology, including rapid assembly of novel combinations of genes, biosensors, and biocomputing. An important requirement for optimal experimental design and predictability when using integrases, however, is the need for multiple enzymes with different
Manteca, Angel; Pelaez, Ana Isabel; del Mar Garcia-Suarez, Maria
A Streptomyces sp. isolated from a patient who had had breast reconstruction after a mastectomy was identified at the species level by comparative sequence analysis of 16S ribosomal DNA (rDNA) and the hypervariable alpha-region of the 16S rDNA....
Také, Akira; Matsumoto, Atsuko; Ōmura, Satoshi; Takahashi, Yōko
Actinomycete strains OM-6519(T) and K04-0144(T) produce the bioactive compounds lactacystin and cyslabdan, respectively. Here, the taxonomic positions of these two strains were determined. The morphological and chemical features of strains OM-6519(T) and K04-0144(T) indicated that they belonged to the genus Streptomyces. Strain OM-6519(T) showed the highest 16S rRNA gene sequence similarities with Streptomyces xanthocidicus NBRC 13469(T) (99.7%), Streptomyces chrysomallus subsp. fumigatus NBRC 15394(T) (99.6%) and Streptomyces aburaviensis NRRL B-2218(T) (99.5%). However, the DNA-DNA relatedness values between strain OM-6519(T) and the three related strains were below 70%. Strain K04-0144(T) showed the highest 16S rRNA gene sequence similarities with Streptomyces corchorusii NBRC 13032(T) (99.4%), Streptomyces olivaceoviridis NBRC 15394(T) (99.4%) and Streptomyces canarius NRRL B-2218(T) (99.3%). However, the DNA-DNA relatedness values between strain K04-0144(T) and the three related strains were also below 70%. Based on morphological, cultural and physiological characteristics and DNA-DNA relatedness data, strains OM-6519(T) and K04-0144(T) should be classified as new species of the genus Streptomyces, for which the names Streptomyces lactacystinicus sp. nov. and Streptomyces cyslabdanicus sp. nov. are proposed. The type strain of S. lactacystinicus is OM-6519(T) (=NBRC 110082(T), DSM 43136(T)). The type strain of S. cyslabdanicus is K04-0144(T) (=NBRC 110081(T), DSM 42135(T)).
Riley, Sean P; Macaluso, Kevin R; Martinez, Juan J
Genetic manipulation of obligate intracellular bacteria of the genus Rickettsia is currently undergoing a rapid period of change. The development of viable genetic tools, including replicative plasmids, transposons, homologous recombination, fluorescent protein-encoding genes, and antibiotic selectable markers has provided the impetus for future research development. This unit is designed to coalesce the basic methods pertaining to creation of genetically modified Rickettsia. The unit describes a series of methods, from inserting exogenous DNA into Rickettsia to the final isolation of genetically modified bacterial clones. Researchers working towards genetic manipulation of Rickettsia or similar obligate intracellular bacteria will find these protocols to be a valuable reference.
Xu, Bo; Chen, Wei; Wu, Zhi-ming; Long, Yue; Li, Kun-tai
Phytopathogenic fungi would induce a variety of plant diseases, resulting in a severe reduction of agricultural output. However, the current plant disease control is mainly dependent on the environmentally and healthily hazardous chemical fungicides. Thus, the present work aimed to isolate an effective antagonistic microorganism against various soilborne phytopathogenic fungi. By dual culture with Rhizoctonia solani, a novel Streptomyces specie, Streptomyces sp. N2, was screened out from a total of 167 isolated actinomycetes, which displayed a strong inhibitory effect on R. solani (26.85 ± 1.35 mm of inhibition zone diameter). By means of macroporous resin and silica gel column chromatography coupled with preparative HPLC, an antifungal metabolite (3-methyl-3,5-amino-4-vinyl-2-pyrone, C6H7O2N) was isolated and purified from Streptomyces sp. N2. The bioassay results showed that the purified antifungal metabolite could not only possess a broad-spectrum inhibitory effect on a range of plant pathogenic fungi in vitro (e.g., R. solani, Pyricularia grisea, Fusarium oxysporum f. sp. niveum, F. oxysporum f. sp. vasinfectum, Penicillium italicum, and Colletotrichum gloeosporioides), but also had a significantly effective in vivo biocontrol efficacy on grape fruits anthracnose caused by C. gloeosporioides. Microscopic observation indicated that the antifungal metabolite from Streptomyces sp. N2 would exert its antimicrobial activity by disorganizing the cytoplasmic organelles of phytopathogenic fungi. The above results suggested that Streptomyces sp. N2 was one of promising fungicide for biocontrol of fungal plant diseases, especially due to its broad-spectrum and effective antagonist on various plant pathogens.
Matsuo, Hirotaka; Kondo, Yoshiyuki; Kawasaki, Takashi; Tokuyama, Shinji; Imamura, Nobutaka
An inhibitor of 3T3-L1 adipocyte differentiation was isolated from Streptomyces sp. TK08330 and identified by spectroscopy as the 18-membered macrolide borrelidin. Treatment with 1.0 μM borrelidin suppressed intracellular lipid accumulation by 80% and inhibited the expression of adipocyte-specific genes. Borrelidin suppressed the mRNA expression of two master regulators of adipocyte differentiation, peroxisome proliferator-activated receptor gamma (PPARγ) and CCAAT/enhancer binding protein (C/EBPα). Studies on well-known upstream regulators of PPARγ revealed that borrelidin down-regulated C/EBPδ mRNA expression but did not affect expression of C/EBPβ. Borrelidin increased mRNA expression of negative regulators of differentiation such as GATA-binding protein (GATA) 3, Krüppel-like factor (KLF) 3 and KLF7, as well as positive regulators, KLF4, KLF6 and KLF15, at early stages of differentiation. To elucidate a primary mediator of borrelidin differentiation inhibitory activity, small interfering RNA (siRNA) transfection experiments were performed. The mRNA expression of PPARγ, which was down-regulated by borrelidin, was not changed by KLF3 and KLF7 siRNA treatment. In contrast, expression of PPARγ in GATA-3 siRNA-treated cells was not significantly different from that of control siRNA-treated cells. Borrelidin significantly inhibited lipid accumulation in control siRNA-treated cells, and treatment with GATA-3 siRNA slightly reduced the inhibitory effect of borrelidin. These results indicate that borrelidin inhibited adipocyte differentiation partially via GATA-3.
Full Text Available Identifying new sources for small molecule discovery is necessary to help mitigate the continuous emergence of antibiotic-resistance in pathogenic microbes. Recent studies indicate that one potentially rich source of novel natural products is Actinobacterial symbionts associated with social and solitary Hymenoptera. Here we test this possibility by examining two species of solitary mud dauber wasps, Sceliphron caementarium and Chalybion californicum. We performed enrichment isolations from 33 wasps and obtained more than 200 isolates of Streptomyces Actinobacteria. Chemical analyses of 15 of these isolates identified 11 distinct and structurally diverse secondary metabolites, including a novel polyunsaturated and polyoxygenated macrocyclic lactam, which we name sceliphrolactam. By pairing the 15 Streptomyces strains against a collection of fungi and bacteria, we document their antifungal and antibacterial activity. The prevalence and anti-microbial properties of Actinobacteria associated with these two solitary wasp species suggest the potential role of these Streptomyces as antibiotic-producing symbionts, potentially helping defend their wasp hosts from pathogenic microbes. Finding phylogenetically diverse and chemically prolific Actinobacteria from solitary wasps suggests that insect-associated Actinobacteria can provide a valuable source of novel natural products of pharmaceutical interest.
Figueras, Maria José; Collado, Luis; Levican, Arturo; Perez, Jessica; Solsona, Maria Josep; Yustes, Clara
Nineteen bacteria isolates recovered from shellfish samples (mussels and oysters) showed a new and specific 16S rDNA-RFLP pattern with an Arcobacter identification method designed to recognize all species described up to 2008. These results suggested that they could belong to a new species. ERIC-PCR revealed that the 19 isolates belonged to 3 different strains. The sequence of the 16S rRNA gene of a representative strain (F98-3(T)) showed 97.6% similarity with the closest species Arcobacter marinus followed by Arcobacter halophilus (95.6%) and Arcobacter mytili (94.7%). The phylogenetic analysis with the16S rRNA, rpoB, gyrB and hsp60 genes placed the shellfish strains within the same cluster as the three species mentioned (also isolated from saline habitats) but they formed an independent phylogenetic line. The DDH results between strain F98-3(T) and A. marinus (54.8%±1.05), confirmed that it represents a new species. Several biochemical tests differentiated the shellfish isolates from all other Arcobacter species. Although the new species was different from A. mytili, they shared not only the same habitat (mussels) but also the characteristic of being so far the only Arcobacter species that are simultaneously negative for urea and indoxyl acetate hydrolysis. All results supported the classification of the shellfish strains as a new species, for which the name Arcobacter molluscorum sp. nov. with the type strain F98-3(T) is proposed (=CECT 7696(T)=LMG 25693(T)).
Toka Özer, Türkan; Durmaz, Süleyman; Yula, Erkan
Candida spp. are the most common opportunistic mycosis worldwide. Although Candida albicans is the most common cause of urinary tract infections, the frequency of non-albicans Candida species is increasing with common use of antifungal in the prophylaxis and treatment. This may lead to difficulties in treatment. Antifungal tests should be applied with identification of species for effective treatment. In this study, identification of Candida species isolated from urine culture and investigation of susceptibility of these strains to amphotericin B, flucytosine, fluconazole, voriconazole was aimed. In this study, 58 Candida strains isolated from urine cultures at Osmaniye State Hospital between January 2012 and April 2013 were included. Urine culture and antifungal susceptibility tests were applied. Incidence rate of Candida spp. was determined as C. albicans (56.9%), Candida glabrata (20.6%), Candida tropicalis (10.3%), Candida parapsilosis (7%), Candida krusei (3.4%), Candida kefyr (1.8%). Most of the isolates were susceptible to amphotericin B, flucytosine, fluconazole, voriconazole. Twenty three (39.7%) Candida strains were isolated from internal medical branches and Intensive Care Unit and 12 (20.6%) from the Surgical Medical Branches. C. albicans and C. glabrata species were isolated most frequently as a candiduria factor in this hospital between January 2012 and April 2013. The analysis of antifungal susceptibility profile shows no significant resistance to antifungals. Copyright © 2016 Japanese Society of Chemotherapy and The Japanese Association for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.
May 8, 2013 ... 2Soil, Water and Environmental Research Institute, Agricultural Research Center, 9 Gamaa st., P.O. Box .... The fluorescent-labeled fragments were purified from the ..... Streptomyces spp. isolates from tea plantation soil. Res.
Coaxes, J.D.; Phillips, E.J.P.; Lonergan, D.J.; Jenter, H.; Lovley, D.R.
In an attempt to better understand the microorganisms responsible for Fe(III) reduction in sedimentary environments, Fe(III)-reducing microorganisms were enriched for and isolated from freshwater aquatic sediments, a pristine deep aquifer, and a petroleum-contaminated shallow aquifer. Enrichments were initiated with acetate or toluene as the electron donor and Fe(III) as the electron acceptor. Isolations were made with acetate or benzoate. Five new strains which could obtain energy for growth by dissimilatory Fe(III) reduction were isolated. All five isolates are gram- negative strict anaerobes which grow with acetate as the electron donor and Fe(III) as the electron acceptor. Analysis of the 16S rRNA sequence of the isolated organisms demonstrated that they all belonged to the genus Geobacter in the delta subdivision of the Proteobacteria. Unlike the type strain, Geobacter metallireducens, three of the five isolates could use H2 as an electron donor fur Fe(III) reduction. The deep subsurface isolate is the first Fe(III) reducer shown to completely oxidize lactate to carbon dioxide, while one of the freshwater sediment isolates is only the second Fe(III) reducer known that can oxidize toluene. The isolation of these organisms demonstrates that Geobacter species are widely distributed in a diversity of sedimentary environments in which Fe(III) reduction is an important process.
Latha, Selvanathan; Sivaranjani, Govindhan; Dhanasekaran, Dharumadurai
Among diverse actinobacteria, Streptomyces is a renowned ongoing source for the production of a large number of secondary metabolites, furnishing immeasurable pharmacological and biological activities. Hence, to meet the demand of new lead compounds for human and animal use, research is constantly targeting the bioprospecting of Streptomyces. Optimization of media components and physicochemical parameters is a plausible approach for the exploration of intensified production of novel as well as existing bioactive metabolites from various microbes, which is usually achieved by a range of classical techniques including one factor at a time (OFAT). However, the major drawbacks of conventional optimization methods have directed the use of statistical optimization approaches in fermentation process development. Response surface methodology (RSM) is one of the empirical techniques extensively used for modeling, optimization and analysis of fermentation processes. To date, several researchers have implemented RSM in different bioprocess optimization accountable for the production of assorted natural substances from Streptomyces in which the results are very promising. This review summarizes some of the recent RSM adopted studies for the enhanced production of antibiotics, enzymes and probiotics using Streptomyces with the intention to highlight the significance of Streptomyces as well as RSM to the research community and industries.
Zhi, Yan; Wu, Qun; Du, Hai; Xu, Yan
Streptomyces spp. producing geosmin have been regarded as the most frequent and serious microbial contamination causing earthy off-flavor in Chinese liquor. It is therefore necessary to control the Streptomyces community during liquor fermentation. Biological control, using the native microbiota present in liquor making, appears to be a better solution than chemical methods. The objective of this study was to isolate native microbiota antagonistic toward Streptomyces spp. and then to evaluate the possible action mode of the antagonists. Fourteen Bacillus strains isolated from different Daqu (the fermentation starter) showed antagonistic activity against Streptomyces sampsonii, which is one of the dominant geosmin producers. Bacillus subtilis 2-16 and Bacillus amyloliquefaciens 1-45 from Maotai Daqu significantly inhibited the growth of S. sampsonii by 57.8% and 84.3% respectively, and effectively prevented the geosmin production in the simulated fermentation experiments (inoculation ratio 1:1). To probe the biocontrol mode, the ability of strain 2-16 and 1-45 to produce antimicrobial metabolites and to reduce geosmin in the fermentation system was investigated. Antimicrobial substances were identified as lipopeptides by ultra-performance liquid chromatography tandem electrospray ionization/quadrupole-time-of-flight mass spectrometry (UPLC-ESI/Q-TOF MS) and in vitro antibiotic assay. In addition, strains 2-16 and 1-45 were able to remove 45% and 15% of the geosmin respectively in the simulated solid-state fermentation. This study highlighted the potential of biocontrol, and how the use of native Bacillus species in Daqu could provide an eco-friendly method to prevent growth of Streptomyces spp. and geosmin contamination in Chinese liquor fermentation. Copyright © 2016 Elsevier B.V. All rights reserved.
Hsiao, Nai-hua; Kirby, Ralph
DNA/DNA microarray hybridization was used to compare the genome content of Streptomyces avermitilis, Streptomyces cattleya, Streptomyces maritimus and Kitasatospora aureofaciens with that of Streptomyces coelicolor A3(2). The array data showed an about 93% agreement with the genome sequence data ava
Streptomyces avermitilis contains a unique restriction system that restricts plasmid DNA containing N6-methyladenine or 5-methylcytosine. Shuttle vectors isolated from Escherichia coli RR1 or plasmids isolated from modification-proficient Streptomyces spp. cannot be directly introduced into S. avermitilis. This restriction barrier can be overcome by first transferring plasmids into Streptomyces lividans or a modification-deficient E. coli strain and then into S. avermitilis. The transformatio...
Weber, D J; Saviteer, S M; Rutala, W A; Thomann, C A
To determine the clinical significance of blood isolates of Bacillus, we reviewed all blood cultures obtained at North Carolina Memorial Hospital between 1981 and 1985. Over the five-year study period the number of patients (incidence per 10,000 hospital admissions) from whom Bacillus was isolated increased from 4.97 in 1981 to 12.5 in 1985. The incidence per 1,000 blood cultures also increased from 1.12 in 1981 to 2.33 in 1985. Review of the medical records of 78 of the 95 patients (82%) with positive cultures allowed retrospective classification of five isolates (6.4%) as clinically significant, 33 isolates (42.3%) as possibly significant, and 40 isolates (51.3%) as nonsignificant. Underlying diseases in patients with clinically significant Bacillus bacteremia included burn trauma in two, leukemia in one, carcinoma in one, and gastrointestinal hemorrhage in one. All isolates judged to be clinically significant and the majority of possibly significant isolates were B cereus. We conclude that the isolation of Bacillus species from blood cultures is clinically significant in 5% to 10% of cases, that the incidence of Bacillus bacteremia is increasing, and that burn trauma should be added to the list of conditions known to predispose to clinically significant Bacillus bacteremia.
Xiang LI; Yi Nan ZHENG; Wen Han LIN; Isabel SATTLER
A novel butanolide, named virginaebutanolide F (1), was isolated from the lyophilized culture broth of Streptomyces sp., along with a known compound virginaebutanolide C (2). Their structures including the stereochemistry were elucidated on the basis of extensive 1D and 2D NMR as well as HRESI-MS and CD spectroscopic analysis.
Full Text Available Aim: The aim of the present work was to ascertain the bacterial flora causing pyometra in female dogs and their antibiotic sensitivity. Materials and Methods: A study was conducted to determine the antibiogram of bacterial species isolated from 20 female dogs diagnosed with pyometra. The vaginal discharge was collected by sterile swab and streaked smoothly over Mueller Hinton medium and sensitivity towards antibiotics was determined by measuring the zone of inhibition using a Hi-media scale. Results: The antobiogram showed that Gentamicin was the most sensitive (85% antibiotic followed by Enrofloxacin, Ciprofloxacin and Amoxicillin (65%, 65% and 55%, respectively. The isolates were most resistant to Oxytetracycline (85% followed by Tetracycline, Ampicillin, Chloramphenicol, Cloxacillin and Erythromycin (80%, 80%, 75%, 70% and 70%, respectively. Conclusion: Gentamicin was found to be most effective antibiotic against the bacterial species isolated from canine pyometra. [Vet World 2013; 6(8.000: 546-549
DNA/DNA microarray hybridization was used to compare the genome content of Streptomyces avermitilis, Streptomyces cattleya, Streptomyces maritimus and Kitasatospora aureofaciens with that of Streptomyces coelicolor A3(2). The array data showed an about 93% agreement with the genome sequence data available for S. avermitilis and also showed a number of trends in the genome structure for Streptomyces and closely related Kitasatospora. A core central region was well conserved, which might be pre...
Doralyn S Dalisay
Full Text Available Representatives of the genus Streptomyces from terrestrial sources have been the focus of intensive research for the last four decades because of their prolific production of chemically diverse and biologically important compounds. However, metabolite research from this ecological niche had declined significantly in the past years because of the rediscovery of the same bioactive compounds and redundancy of the sample strains. More recently, a new picture has begun to emerge in which marine-derived Streptomyces bacteria have become the latest hot spot as new source for unique and biologically active compounds. Here, we investigated the marine sediments collected in the temperate cold waters from British Columbia, Canada as a valuable source for new groups of marine-derived Streptomyces with antimicrobial activities. We performed culture dependent isolation from 49 marine sediments samples and obtained 186 Streptomyces isolates, 47 of which exhibited antimicrobial activities. Phylogenetic analyses of the active isolates resulted in the identification of four different clusters of bioactive Streptomyces including a cluster with isolates that appear to represent novel species. Moreover, we explored whether these marine-derived Streptomyces produce new secondary metabolites with antimicrobial properties. Chemical analyses revealed structurally diverse secondary metabolites, including four new antibacterial novobiocin analogues. We conducted structure-activity relationships (SAR studies of these novobiocin analogues against methicillin-resistant Staphylococcus aureus (MRSA. In this study, we revealed the importance of carbamoyl and OMe moieties at positions 3" and 4" of novobiose as well as the hydrogen substituent at position 5 of hydroxybenzoate ring for the anti-MRSA activity. Changes in the substituents at these positions dramatically impede or completely eliminate the inhibitory activity of novobiocins against MRSA.
Grishko, V. N.; Syshchikova, O. V.
The contents of differently mobile heavy metal compounds and their influence on the formation of microbial cenoses (particularly, streptomyces communities) in technogenically disturbed soils are considered. Elevated concentrations of mobile Cu, Zn, Ni, Cd, and Fe compounds are shown to determine structural-functional changes in microbial cenoses that are displayed in a decreasing number of microorganisms and a narrower spectrum of the streptomyces species. Some specific features of the formation of streptomyces communities in technogenic soils were revealed on the basis of the analysis of their species structure with the use of the Margalef, Berger-Parker, and Sorensen indices of biodiversity.
Background: The growing problem of environmental pollution caused by synthetic plastics has led to the search for alternative materials such as biodegradable plastics. Of the biopolymers presently under development, starch/natural rubber is one promising alternative. Several species of bacteria and fungi are capable of degrading natural rubber and many can degrade starch. Results: Streptomyces coelicolor CH13 was isolated from soil according to its ability to produce translucent halos on a mi...
Human, Zander R.; Moon, Kyuho; Bae, Munhyung; de Beer, Z. Wilhelm; Cha, Sangwon; Wingfield, Michael J.; Slippers, Bernard; Oh, Dong-Chan; Venter, Stephanus N.
Common saprophytic fungi are seldom present in Protea infructescences, which is strange given the abundance of mainly dead plant tissue in this moist protected environment. We hypothesized that the absence of common saprophytic fungi in Protea infructescences could be due to a special symbiosis where the presence of microbes producing antifungal compounds protect the infructescence. Using a culture based survey, employing selective media and in vitro antifungal assays, we isolated antibiotic producing actinomycetes from infructescences of Protea repens and P. neriifolia from two geographically separated areas. Isolates were grouped into three different morphological groups and appeared to be common in the Protea spp. examined in this study. The three groups were supported in 16S rRNA and multi-locus gene trees and were identified as potentially novel Streptomyces spp. All of the groups had antifungal activity in vitro. Streptomyces sp. Group 1 had inhibitory activity against all tested fungi and the active compound produced by this species was identified as fungichromin. Streptomyces spp. Groups 2 and 3 had lower inhibition against all tested fungi, while Group 3 showed limited inhibition against Candida albicans and Sporothrix isolates. The active compound for Group 2 was also identified as fungichromin even though its production level was much lower than Group 1. The antifungal activity of Group 3 was linked to actiphenol. The observed antifungal activity of the isolated actinomycetes could contribute to protection of the plant material against common saprophytic fungi, as fungichromin was also detected in extracts of the infructescence. The results of this study suggest that the antifungal Streptomyces spp. could play an important role in defining the microbial population associated with Protea infructescences. PMID:27853450
Full Text Available Common saprophytic fungi are seldom present in Protea infructescences, which is strange given the abundance of mainly dead plant tissue in this moist protected environment. We hypothesized that the absence of common saprophytic fungi in Protea infructescences could be due to a special symbiosis where the presence of microbes producing antifungal compounds protect the infructescence. Using a culture based survey, employing selective media and in vitro antifungal assays, we isolated antibiotic producing actinomycetes from infructescences of Protea repens and P. neriifolia from two geographically separated areas. Isolates were grouped into three different morphological groups and appeared to be common in the Protea spp. examined in this study. The three groups were supported in 16S rRNA and multi-locus gene trees and were identified as potentially novel Streptomyces spp. All of the groups had antifungal activity in vitro. Streptomyces sp. Group 1 had inhibitory activity against all tested fungi and the active compound produced by this species was identified as fungichromin. Streptomyces spp. Groups 2 and 3 had lower inhibition against all tested fungi, while Group 3 showed limited inhibition against Candida albicans and Sporothrix isolates. The active compound for Group 2 was also identified as fungichromin even though its production level was much lower than Group 1. The antifungal activity of Group 3 was linked to actiphenol. The observed antifungal activity of the isolated actinomycetes could contribute to protection of the plant material against common saprophytic fungi, as fungichromin was also detected in extracts of the infructescence. The results of this study suggest that the antifungal Streptomyces spp. could play an important role in defining the microbial population associated with Protea infructescences.
Luis E. Espinoza
Full Text Available Streptomyces are common soil bacteria that produce secondary metabolites, including several antibiotics; however, the characteristics of marine Streptomyces are largely unknown. Sediment samples were taken from 3 sites in the Laguna Madre to isolate marine Streptomyces. Sediment was diluted, spread onto synthetic seawater media to estimate the total bacterial density of the samples and spread onto starch casein agar to isolate Streptomyces. Isolated Streptomyces were tested for salinity tolerance and optimal growth pH. Isolates were assayed using API 20EÂ® test strips and BIOLOGâ¢ plates to construct biochemical profiles and assess nutrient utilization abilities of the bacteria, respectively. Individual Streptomyces were tested for the ability to inhibit the growth of other isolated Streptomyces (i.e., interference competition and putatively identified by DNA sequencing. Results showed that there was no significant difference in microbial density in sediments from the 3 sampling sites. Eleven (11 Streptomyces pure cultures were obtained in total; most tolerated salinity up to 60 ppt and grew optimally at pH 7.5. Biochemical profile comparisons showed that the Streptomyces were only at least 74% similar; most (8/11 were >90% similar. Isolates could use between 87-95 carbon sources. Three (3 isolates displayed interference toward other isolates. Ten (10 isolates were identified as Streptomyces griseus by DNA sequencing. Laguna Madre Streptomyces organisms display some diverse characteristics with regards to their halotolerance, biochemical profiles, carbon source utilization and inhibition toward other organisms. Further investigations may yield greater understanding of these organisms in this and other marine environments and may be a reservoir of novel microorganisms and secondary metabolites.
Katz, L; Chiang, S J; Tuan, J S; Zablen, L B
A bacteriophage, designated phi C69, isolated from a culture of Saccharopolyspora erythraea was characterized. The phage propagates on Sac. erythraea NRRL 2338 but does not infect 10 Streptomyces or 3 Micromonospora species tested. It infects Sac. erythraea NRRL 2359 but does not produce infectious phage particles in this host. phi C69 is approximately 40 kb in length and contains cohesive ends. A cos fragment containing ligated phage DNA ends was cloned in Escherichia coli. Restriction maps of the phage DNA and the cos fragment for several enzymes are shown. Transfection of both Sac. erythraea and Streptomyces lividans with phi C69 resulted in approximately equal titres of infectious phage particles produced from approximately the same number of regenerating cells. Transfection of Sac. erythraea with DNA from Streptomyces phages SH10 and KC404 also resulted in the production of infectious phage particles. The basis for differences among hosts in susceptibility to infection by various actinophages is discussed.
Ready, J S; Sampaio, I; Schneider, H; Vinson, C; Dos Santos, T; Turner, G F
Laboratory mate choice experiments have confirmed species status for cichlid fish in the African Great Lakes that differ in colour and little else. Colour differences between allopatric populations of the South American cichlid genus Apistogramma are known for many species, yet the status of such populations has not been previously tested. Analysis of the genetic relationships and mate choice characteristics of populations previously described as Apistogramma caetei from eastern Amazonia indicates genetic differentiation into at least three allopatric lineages, which also show strong prezygotic isolation through female mate choice, confirming them as Biological species. If future studies confirm that this result is indicative of a general trend, the species richness of the South American cichlid fishes may presently be seriously underestimated.
Labeda, D P; Lyons, A J
DNA relatedness among 28 putative strains of Streptomyces ipomoea from geographically diverse locations and the type strain, NRRL B-12321, was determined spectrophotometrically. The data confirm that these 28 strains are not closely related genetically to the plant-pathogenic species Streptomyces scabies (39% DNA relatedness) or Streptomyces acidiscabies (17% DNA relatedness) or any other major blue-spored Streptomyces species (less than 30% DNA relatedness). Of the 29 strains examined, 4 could be clearly distinguished from S. ipomoea on the basis of morphological criteria, i.e., they had gray rather than blue spores and produced melanin pigment, and their low DNA relatedness to authentic S. ipomoea strains confirmed their original misidentification. The remaining 25 S. ipomoea strains exhibited high DNA relatedness among themselves (76 to 100% homology), even though they had been isolated in different locations throughout the United States and Japan. The avirulent type strain, NRRL B-12321, exhibited slightly lower DNA relatedness with the virulent strains of S. ipomoea (85% average DNA relatedness) than was observed among the virulent strains (average of 96% DNA relatedness).
Tsuzukibashi, Osamu; Uchibori, Satoshi; Kobayashi, Taira; Umezawa, Koji; Mashimo, Chiho; Nambu, Takayuki; Saito, Masanori; Hashizume-Takizawa, Tomomi; Ochiai, Tomoko
Rothia dentocariosa and Rothia mucilaginosa which are Gram-positive bacteria are part of the normal flora in the human oral cavity and pharynx. Furthermore, Rothia aeria, which was first isolated from air samples in the Russian space station Mir, is predicted to be an oral inhabitant. Immunocompromised patients are often infected by these organisms, leading to various systemic diseases. The involvement of these organisms in oral infections has attracted little attention, and their distribution in the oral cavity has not been fully clarified because of difficulties in accurately identifying these organisms. A suitable selective medium for oral Rothia species, including R. aeria, is necessary to assess the veritable prevalence of these organisms in the oral cavity. To examine the bacterial population in the oral cavity, a novel selective medium (ORSM) was developed for isolating oral Rothia species in this study. ORSM consists of tryptone, sodium gluconate, Lab-Lemco powder, sodium fluoride, neutral acriflavin, lincomycin, colistin, and agar. The average growth recovery of oral Rothia species on ORSM was 96.7% compared with that on BHI-Y agar. Growth of other representative oral bacteria, i.e. genera Streptococcus, Actinomyces, Neisseria, and Corynebacterium, was remarkably inhibited on the selective medium. PCR primers were designed based on partial sequences of the 16S rDNA genes of oral Rothia species. These primers reacted to each organism and did not react to other non-oral Rothia species or representative oral bacteria. These results indicated that these primers are useful for identifying oral Rothia species. A simple multiplex PCR procedure using these primers was a reliable method of identifying oral Rothia species. The proportion of oral Rothia species in saliva samples collected from 20 subjects was examined by culture method using ORSM. Rothia dentocariosa, Rothia mucilaginosa, and R. aeria accounted for 1.3%, 5.9%, and 0.8% of the total cultivable
Poulsen, Michael; Oh, Dong-Chan; Clardy, Jon;
Identifying new sources for small molecule discovery is necessary to help mitigate the continuous emergence of antibiotic-resistance in pathogenic microbes. Recent studies indicate that one potentially rich source of novel natural products is Actinobacterial symbionts associated with social...... of these isolates identified 11 distinct and structurally diverse secondary metabolites, including a novel polyunsaturated and polyoxygenated macrocyclic lactam, which we name sceliphrolactam. By pairing the 15 Streptomyces strains against a collection of fungi and bacteria, we document their antifungal...... and antibacterial activity. The prevalence and anti-microbial properties of Actinobacteria associated with these two solitary wasp species suggest the potential role of these Streptomyces as antibiotic-producing symbionts, potentially helping defend their wasp hosts from pathogenic microbes. Finding...
Deshpande, Neelima; Choubey, Prachi; Agashe, Manasi
A species of Streptomyces, Streptomyces ginsengisoli, a river isolate, was evaluated for production of an enzyme, L-asparaginase, with multiple functions mainly anticancer activity. The actinomycete was subjected to submerged fermentation by "shake flask" method. The quantity of L-asparaginase produced was estimated as 3.23 μ mol/mL/min. The effect of various culture conditions on L-asparaginase production was studied by adopting a method of variation in one factor at a time. Of the various conditions tested, glucose (followed by starch) and peptone served as good carbon and nitrogen sources, respectively, for maximal production of enzyme at pH 8. The temperature of 30 °C and an incubation period of 5 days with 0.05 g% asparagine concentration were found to be optimum for L-asparaginase production.
Full Text Available A species of Streptomyces, Streptomyces ginsengisoli, a river isolate, was evaluated for production of an enzyme, L-asparaginase, with multiple functions mainly anticancer activity. The actinomycete was subjected to submerged fermentation by “shake flask” method. The quantity of L-asparaginase produced was estimated as 3.23 μmol/mL/min. The effect of various culture conditions on L-asparaginase production was studied by adopting a method of variation in one factor at a time. Of the various conditions tested, glucose (followed by starch and peptone served as good carbon and nitrogen sources, respectively, for maximal production of enzyme at pH 8. The temperature of 30°C and an incubation period of 5 days with 0.05 g% asparagine concentration were found to be optimum for L-asparaginase production.
Said Mohamed Daboor; Amany Mohamed Haroon; Neven Abd Elfatah Esmael; Slah Ibrahem Hanona
Objective:To find the best actinomycete that has potential application value in the heavy metal remediation due to its special morphological and physiological metabolism. Methods: In some areas of River Nile, Egypt, a total of 67 actinomycete isolates (17 isolates from surface water and 50 from sediment) were identified. In addition, the studied area was characterized by a large amount of submerged macrophyte species Ceratophyllum demersum, one free floating species Eichhornia crassipes and two emergent species Polygonum tomentosum and Saccharum spontaneum with the highest biomass production values. Many methods are used in this research like qualitative evaluation of heavy metals, minimum inhibitory concentration of heavy metal determination, metal binding assay, heavy metal assessment, etc. Results: Many actinomycetes isolates were isolated from River Nile, Egypt, the absorbent efficiency of one isolate Streptomyces chromofuscusK101 showed the most efficient metal binding activity. The adsorption process of Zn2+, Pb2+and Fe2+single or mixture metal ions was investigated, where the order of adsorption potential ( Zn2+>Pb2+>Fe2+ ) was observed in single metal reaction. The adsorption in mixed metal reactions was the same order as in single-metal ion with a significant decrease in Fe2+and Pb2+adsorption. Conclusions: In conclusion the metal adsorption reactions were very fast, pH dependent and culture age-independent, suggestive of a physicochemical reaction between cell wall components and heavy metal ions. The absorbent removal efficiency was determined as a function of ion concentration, pH and temperature.
Benita Mercy R
Full Text Available Drug resistance by bacterial pathogens becomes a major health problem worldwide. Hence, it is important to search for broad spectrum of antibiotic from natural sources. Marine actinomycetes isolated from marine sediments collected at different sampling sites along the southeast coast of Bay of Bengal, India were investigated for antagonistic activity against selected drug resistant Gram positivebacterial pathogens. All actinomycetes isolates were screened for antibacterial activity against standard drug resistant ATCC strains. The potential isolate which showed higher inhibitory activity against drug resistant pathogens was mass cultured and the ethyl acetate (EA extract of the cell free culture broth was tested for antibacterial activity. The biochemical, morphological and physiological characterisation of the isolate revealed that it was Gram-positive rod, sporulating and produced grey aerial mycelium. The spore chain morphology, and smooth surface morphology showed that it belongs to the genus Streptomyces.Based on Nonomura’s key for classification of Streptomycesand Bergey’s Manual of Determinative Bacteriology, the isolatewas identified as Streptomyces species and designated as Streptomyces sp. VITBRK4.Purification and characterization of EA extract of the isolate by thin layer chromatography (TLC and HPLC-DAD analysis showed the presence of indolo compound along with few other unidentified metabolites. The result of this study showed that the antibacterial activity of the EA extract against drug resistant strains may be due to indolo compound present in the extract.
I WAYAN EKA DHARMAWAN
Full Text Available An exploration study of natural resources soil bacteria antibiotic-producer, Streptomyces spp. was done in two steps. The first step was isolation of Streptomyces and the second involved testing their inhibition activities against five strains diarrheagenic Escherichia coli. Soil samples were collected from ten forest areas in Bali. As many as 55 isolates were collected with various macroscopic dan microscopic characters. Most isolates (eight Streptomyces isolates were collected from forest area in Penulisan, Kintamani (RTK. 20. The diversities of isolates are influenced by environment condition. All Streptomyces isolated were tested against five strains diarrheagenic Escherichia coli to check antibiotic activity for inhibit growth of E. coli. Streptomycine was used as a control. The result showed that the largest inhibition zones of Streptomyces against E. coli strains EHEC, ETEC, EIEC, EPEC and DAEC were produced by Streptomyces PK5 (48,67 ± 0,58 mm, Streptomyces GAA4 (29,00 ± 2,00 mm, Streptomyces GBK3 (42,67 ± 2,08 mm, Streptomyces SkBB5 (29,00 ± 2,65 mm and Streptomyces GM3 (33,67 ± 3,21 mm respectively.
Kim, Ga-Yeon; Jeon, Jae-Sik; Kim, Jae Kyung
Candida spp. is an invasive infectious fungus, a major risk factor that can increase morbidity and mortality in hospitalized patients. In this study, 2,508 Candida spp. were isolated from various clinical specimens collected from university hospitals from July 2011 to October 2014. They were identified in order to determine isolation frequencies and characteristics by specimen, gender, age group, year, season, and month. The strain-specific isolation rate of Candida spp. is in the order of Candida albicans (1,218 strains, 48.56%), Candida glabrata (416 strains, 16.59%), Candida utilis (305 strains, 12.16%), Candida tropicalis (304 strains, 12.12%), and Candida parapsilosis (116 strains, 4.63%) and these five species accounted for more than 94% of the total strains. Of the specimens, Candida spp. were most frequently isolated from urine-catheter, followed by urine-voided, blood, sputum, other, open pus, vaginal discharge, Tip, ear discharge, bronchial aspiration and bile, in that order. Looking at the age distribution, the detection rate of patients in their 60s and older was significantly higher at 75.8% (1,900/2,508). The detection rate of patients in their 20s and younger was shown to be very low at 2.55% (64/2,508). By year, the detection rate of non-albicans Candida spp. showed a tendency to gradually increase each year compared with C. albicans. As isolation of Candida spp. from clinical samples at the specie level can vary depending on characteristics of the patient, sample, season, etc., continual studies are required.
Monson, Rita; Salmond, George P C
Streptomyces coelicolor is a model system for the study of Streptomyces, a genus of bacteria responsible for the production of many clinically important antibiotics. Here we report the genome sequence of ΦCAM, a new S. coelicolor generalized transducing bacteriophage, isolated from a soil sample originating from Lincolnshire, United Kingdom. Many open reading frames within ΦCAM shared high levels of similarity to a prophage from Salinispora tropica and a putative prophage in Streptomyces sp. strain C.
Full Text Available Introduction: Streptomyces species are mycelial, aerobic gram-positive bacteria that are isolated from soil and produce a diverse range of antibiotics. Streptomyces griseus produces the antibiotic, streptomycin and forms spores even in a liquid culture. The gene cluster for the production of Streptomycin antibiotic contains strR gene that encodes StrR, a pathway-specific regulator. Then, this pathway-specific regulator induces transcription of other streptomycin production genes in the gene cluster. The overall aim of this work was rapid isolation and molecular detection of streptomycin-producing Streptomycetes, especially S. griseus, from Iranian soils in order to manipulate them for increased production of streptomycin. Methods: This research used new initiative half-specific medium for isolation of Streptomycetes from natural environments, called FZmsn. The fifty colonies of Streptomyces strains grown on the surface of FZmsn medium isolated from environmental samples were defined on the basis of their morphological characteristics and light microscope studies. A set of primers was designed to detect strR by OLIGO software. Results: In colony-PCR reactions followed by gel electrophoresis, 6 colonies from Streptomyces strains colonies were detected as S. griseus colonies. Conclusion: These native Streptomyces strains will be used for genetic manipulation of S. griseus in order to increase production levels of streptomycin.
Sharova, N Iu
Inhibitors synthesized by the Streptomyces lucensis VKPM AS-1743 and Streptomyces violaceus VKPM AS-1734 strains were studied for their influence on amylases of different origin. The effect of the inhibitors was shown to be different on fungal amylase, pancreatic amylase, and amylase from human blood. It has been found that the studied inhibitors are substances of a pseudooligosaccharide nature and exhibit their activity and stability over a wide range of pH and temperature values. The physico-chemical and biochemical properties of isolated inhibitors were compared with those of known microbial inhibitors of α-glucosidases.
Fathi, Naeimeh; Mohammadi, Rasoul; Tabatabaiefar, Mohammad Amin; Ghahri, Mohammad; Sadrossadati, Seyedeh Zahra
Candida species are the most prevalent cause of invasive fungal infections such as candidemia. Candidemia is a lethal fungal infection among immunocompromised patients worldwide. Main pathogen is Candida albicans but a global shift in epidemiology toward non-albicans species have reported. Species identification is imperative for good management of candidemia as a fatal infection. The aim of the study is to identify Candida spp. obtained from candidemia and determination of mortality rate among this population. The study was performed during February 2014 to March 2015 in Tehran, Iran. Two-hundred and four blood cultures were evaluated for fungal bloodstream infection. Identification of isolates was carried out using phenotypic tests and polymerase chain reaction sequencing technique. Twenty-two out of 204 patients (10.8%) had candidemia. Candida parapsilosis was the most prevalent species (45.4%), followed by C. albicans (31.8%) and Candida glabrata (22.7%). Male to female sex ratio was 8/14. The emergence of resistant strains of Candida species should be considered by physicians to decrease the mortality of this fatal fungal infection by appropriate treatment.
Fathi, Naeimeh; Mohammadi, Rasoul; Tabatabaiefar, Mohammad Amin; Ghahri, Mohammad; Sadrossadati, Seyedeh Zahra
Background: Candida species are the most prevalent cause of invasive fungal infections such as candidemia. Candidemia is a lethal fungal infection among immunocompromised patients worldwide. Main pathogen is Candida albicans but a global shift in epidemiology toward non-albicans species have reported. Species identification is imperative for good management of candidemia as a fatal infection. The aim of the study is to identify Candida spp. obtained from candidemia and determination of mortality rate among this population. Materials and Methods: The study was performed during February 2014 to March 2015 in Tehran, Iran. Two-hundred and four blood cultures were evaluated for fungal bloodstream infection. Identification of isolates was carried out using phenotypic tests and polymerase chain reaction sequencing technique. Results: Twenty-two out of 204 patients (10.8%) had candidemia. Candida parapsilosis was the most prevalent species (45.4%), followed by C. albicans (31.8%) and Candida glabrata (22.7%). Male to female sex ratio was 8/14. Conclusions: The emergence of resistant strains of Candida species should be considered by physicians to decrease the mortality of this fatal fungal infection by appropriate treatment. PMID:27713871
Full Text Available Plants of the Carex genus (Family: Cyperaceae have attracted recent attention as potential food additives due to their high levels of potential bioactive compounds. In this study, the stilbene contents of five unexplored Carex species were investigated: Carex capillacea, Carex hirta, Carex buchananii, Carex cuprina, Carex glauca. High-performance liquid chromatography coupled to mass spectrometry (HPLC–MS and NMR spectroscopy (NMR were used to identify the structures. A novel stilbene oligomer, carexinol A, was isolated together with five previously known stilbenes: resveratrol-diglucoside, miyabenol A and C, kobophenol A and α-viniferin. Furthermore, this is the first report of resveratrol diglucoside in Carex genus.
Pinheiro, Guilherme L; de Azevedo-Martins, Allan C; Albano, Rodolpho M; de Souza, Wanderley; Frases, Susana
The giant snail Achatina fulica is considered an invasive species in most territories in which it was introduced, due to its ability to process a large amount of lignocellulose as a consequence of the presence of a cellulolytic-associated microflora. Streptomyces are well known as crucial agents in the decomposition of complex polymers in soil environments and also as cellulolytic symbionts commonly associated with herbivore insects. Here, we employed a combination of genomic and biochemical tools for a detailed evaluation of the cellulolytic potential of Streptomyces sp. I1.2, an aerobic bacterium isolated from the intestinal lumen of A. fulica in a screening for cellulolytic bacteria. Genomic analysis revealed that the ratio and diversity of CAZy domains and GH families coded by Streptomyces sp. I1.2 are comparable to those present in other highly cellulolytic bacteria. After growth on crystalline cellulose or sugarcane bagasse as sole carbon sources, the functionality of several genes encoding endoglucanases, cellobiohydrolases, xylanases, CBMs, and one β-glucosidase were confirmed by the combination of enzymatic activity measurements, zymography, TLC, and cellulose-binding assays. The endoglucanases secreted by this isolate were stable at 50 °C and exhibited activity over a broad pH range between 4.0 and 8.0. The endoglucanases and cellobiohydrolases secreted by Streptomyces sp. I1.2 exhibited specific activities that were similar to the levels present in a commercial cellulase preparation from Trichoderma reesei, while I1.2 xylanase levels were even 350 % higher. The results presented here show that Streptomyces sp. I1.2 is promising for future biotechnological applications, since it is able to produce endoglucanases, cellobiohydrolases, and xylanases in appreciable amounts when grown on a low-cost residue such as sugarcane bagasse.
Clothier, Kristin A; Kinyon, Joann M; Frana, Timothy S; Naberhaus, Nadine; Bower, Leslie; Strait, Erin L; Schwartz, Kent
Typhlocolitis and dysentery due to Brachyspira hyodysenteriae infection represent an economically important disease syndrome in growing pigs. Largely disappearing from U.S. swine herds in the late 1990 s and early 2000s, Brachyspira-associated disease and bacterial isolation from swine with clinical disease has increased in the last several years, and non-B. hyodysenteriae isolates are commonly identified. Antimicrobial resistance has been demonstrated in Brachyspira spp. isolates from Europe and Asia, and may be the reason for the resurgence in U.S. herds. Seventy-nine clinical isolates identified at the Iowa State University Veterinary Diagnostic Lab were tested with multiple polymerase chain reaction assays to establish species identity, and evaluated for minimum inhibitory concentrations (MICs) using an agar dilution method against lincomycin, gentamicin, valnemulin, tiamulin, salinomycin, and carbadox. Only 38.0% of isolates could be confirmed as the known pathogens B. hyodysenteriae (30.4%) or Brachyspira pilosicoli (7.6%). Twenty of the 79 isolates (25.3%) were identified as Brachyspira murdochii, and 13.9% could not be identified to species. The MIC values were consistently high against lincomycin and moderately high against gentamicin. The remaining antimicrobials had MICs that were at the low end of the test ranges. Brachyspira murdochii and Brachyspira spp. had significantly greater MIC values against several of these drugs than other Brachyspira spp. examined. The increased incidence of these less definitively characterized Brachyspira species with increased MIC values to commonly prescribed antimicrobials may, at least in part, explain the increased prevalence and severity of this disease complex in recent years. Further research is necessary to understand these changes.
Full Text Available Gram positive bacteria produce cell envelope macroamphiphile glycopolymers, i.e. lipoteichoic acids or lipoglycans, whose functions and biosynthesis are not yet fully understood. We report for the first time a detailed structure of lipoteichoic acid isolated from a Streptomyces species, i.e. Streptomyces hygroscopicus subsp. hygroscopicus NRRL 2387T. Chemical, MS and NMR analyses revealed a polyglycerolphosphate backbone substituted with α-glucosaminyl and α-N-acetyl-glucosaminyl residues but devoid of any amino-acid substituent. This structure is very close, if not identical, to that of the wall teichoic acid of this organism. These data not only contribute to the growing recognition that lipoteichoic acid is a cell envelope component of gram positive Actinobacteria but also strongly support the recently proposed hypothesis of an overlap between the pathways of lipoteichoic acid and wall teichoic acid synthesis in these bacteria. S. hygroscopicus lipoteichoic acid induced signalling by human innate immune receptor TLR2, confirming its role as a microbe-associated molecular pattern. Its activity was partially dependant on TLR1, TLR6 and CD14. Moreover, it stimulated TNF-α and IL-6 production by a human macrophage cell line to an extent similar to that of Staphylococcus aureus lipoteichoic acid. These results provide new clues on lipoteichoic acid structure/function relationships, most particularly on the role of the polyglycerolphosphate backbone substituents.
Lee, Learn-Han; Zainal, Nurullhudda; Azman, Adzzie-Shazleen; Eng, Shu-Kee; Goh, Bey-Hing; Yin, Wai-Fong; Ab Mutalib, Nurul-Syakima; Chan, Kok-Gan
The aim of this study was to isolate and identify Actinobacteria from Malaysia mangrove forest and screen them for production of antimicrobial secondary metabolites. Eighty-seven isolates were isolated from soil samples collected at 4 different sites. This is the first report to describe the isolation of Streptomyces, Mycobacterium, Leifsonia, Microbacterium, Sinomonas, Nocardia, Terrabacter, Streptacidiphilus, Micromonospora, Gordonia, and Nocardioides from mangrove in east coast of Malaysia. Of 87 isolates, at least 5 isolates are considered as putative novel taxa. Nine Streptomyces sp. isolates were producing potent antimicrobial secondary metabolites, indicating that Streptomyces isolates are providing high quality metabolites for drug discovery purposes. The discovery of a novel species, Streptomyces pluripotens sp. nov. MUSC 135(T) that produced potent secondary metabolites inhibiting the growth of MRSA, had provided promising metabolites for drug discovery research. The biosynthetic potential of 87 isolates was investigated by the detection of polyketide synthetase (PKS) and nonribosomal polyketide synthetase (NRPS) genes, the hallmarks of secondary metabolites production. Results showed that many isolates were positive for PKS-I (19.5%), PKS-II (42.5%), and NRPS (5.7%) genes, indicating that mangrove Actinobacteria have significant biosynthetic potential. Our results highlighted that mangrove environment represented a rich reservoir for isolation of Actinobacteria, which are potential sources for discovery of antimicrobial secondary metabolites.
Full Text Available The aim of this study was to isolate and identify Actinobacteria from Malaysia mangrove forest and screen them for production of antimicrobial secondary metabolites. Eighty-seven isolates were isolated from soil samples collected at 4 different sites. This is the first report to describe the isolation of Streptomyces, Mycobacterium, Leifsonia, Microbacterium, Sinomonas, Nocardia, Terrabacter, Streptacidiphilus, Micromonospora, Gordonia, and Nocardioides from mangrove in east coast of Malaysia. Of 87 isolates, at least 5 isolates are considered as putative novel taxa. Nine Streptomyces sp. isolates were producing potent antimicrobial secondary metabolites, indicating that Streptomyces isolates are providing high quality metabolites for drug discovery purposes. The discovery of a novel species, Streptomyces pluripotens sp. nov. MUSC 135T that produced potent secondary metabolites inhibiting the growth of MRSA, had provided promising metabolites for drug discovery research. The biosynthetic potential of 87 isolates was investigated by the detection of polyketide synthetase (PKS and nonribosomal polyketide synthetase (NRPS genes, the hallmarks of secondary metabolites production. Results showed that many isolates were positive for PKS-I (19.5%, PKS-II (42.5%, and NRPS (5.7% genes, indicating that mangrove Actinobacteria have significant biosynthetic potential. Our results highlighted that mangrove environment represented a rich reservoir for isolation of Actinobacteria, which are potential sources for discovery of antimicrobial secondary metabolites.
Zhang, Renwen; Han, Xiaoxue; Xia, Zhanfeng; Luo, Xiaoxia; Wan, Chuanxing; Zhang, Lili
A novel actinomycete strain, designated TRM 49605(T), was isolated from a desert soil sample from Lop Nur, Xinjiang, north-west China, and characterised using a polyphasic taxonomic approach. The strain exhibited antifungal activity against the following strains: Saccharomyces cerevisiae, Curvularia lunata, Aspergillus flavus, Aspergillus niger, Fusarium oxysporum, Penicillium citrinum, Candida albicans and Candida tropicalis; Antibacterial activity against Bacillus subtilis, Staphylococcus epidermidis and Micrococcus luteus; and no antibacterial activity against Escherichia coli. Phylogenetic analysis based on 16S rRNA gene sequences affiliated strain TRM 49605(T) to the genus Streptomyces. Strain TRM 49605(T) shows high sequence similarities to Streptomyces roseolilacinus NBRC 12815(T) (98.62 %), Streptomyces flavovariabilis NRRL B-16367(T) (98.45 %) and Streptomyces variegatus NRRL B-16380(T) (98.45 %). Whole cell hydrolysates of strain TRM 49605(T) were found to contain LL-diaminopimelic acid as the diagnostic diamino acid and galactose, glucose, xylose and mannose as the major whole cell sugars. The major fatty acids in strain TRM 49605(T) were identified as iso C16:0, anteiso C15:0, C16:0 and Summed Feature 5 as defined by MIDI. The main menaquinones were identified as MK-9(H4), MK-9(H6), MK-9(H8) and MK-10(H6). The polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine, phosphatidylinositol and phosphatidylinositol mannoside. The G+C content of the genomic DNA was determined to be 71.2 %. The DNA-DNA relatedness between strain TRM 49605(T) and the phylogenetically related strain S. roseolilacinus NBRC 12815(T) was 60.12 ± 0.06 %, which is lower than the 70 % threshold value for delineation of genomic prokaryotic species. Based on the phenotypic, chemotaxonomic and phylogenetic data, strain TRM 49605(T) (=CCTCC AA2015026(T) = KCTC 39666(T)) should be designated as the type strain of a novel species of the genus
Zhao, Fei; Qin, Yu-Hua; Zheng, Xin; Zhao, Hong-Wei; Chai, Dong-Yan; Li, Wei; Pu, Ming-Xiang; Zuo, Xing-Sheng; Qian, Wen; Ni, Ping; Zhang, Yong; Mei, Han; He, Song-Tao
The genus Streptomyces is a widespread genus within the phylum Actinobacteria and has been isolated from various environments worldwide. However, little is known about whether biogeography affects distributional pattern of Streptomyces in salty environments. Such information is essential for understanding the ecology of Streptomyces. Here we analyzed four house-keeping genes (16S rRNA, rpoB, recA and atpD) and salty-tolerance related genes (ectA-ectD) of 38 Streptomyces strains isolated from saline environments in Yunnan and Xinjiang Provinces of western China. The obtained Streptomyces strains were classified into three operational taxonomic units, each comprising habitat-specific geno- and ecotype STs. In combination with expressional variations of salty-tolerance related genes, the statistical analyses showed that spatial distance and environmental factors substantially influenced Streptomyces distribution in saline environments: the former had stronger influence at large spatial scales (>700 km), whereas the latter was influential at large (>700 km) and small spatial scales (ectoine and hydroxyectoine than strains from salt mines, which could help them resist to salinity in the hypersaline environments. PMID:27596681
Anti-phytopathogen potential of endophytic actinobacteria isolated from tomato plants (Lycopersicon esculentum) in southern Brazil, and characterization of Streptomyces sp. R18(6), a potential biocontrol agent.
de Oliveira, Margaroni Fialho; da Silva, Mariana Germano; Van Der Sand, Sueli T
Tomato plants (Lycopersicon esculentum) are highly susceptible to phytopathogen attack. The resulting intensive application of pesticides on tomato crops can affect the environment and health of humans and animals. The objective of this study was to select potential biocontrol agents among actinobacteria from tomato plants, in a search for alternative phytopathogen control. We evaluated 70 endophytic actinobacteria isolated from tomato plants in southern Brazil, testing their antimicrobial activity, siderophore production, indoleacetic acid production, and phosphate solubility. The actinomycete isolate with the highest antimicrobial potential was selected using the agar-well diffusion method, in order to optimize conditions for the production of compounds with antimicrobial activity. For this study, six growth media (starch casein-SC, ISP2, Bennett's, Sahin, Czapek-Dox, and TSB), three temperatures (25 degrees C, 30 degrees C, and 35 degrees C) and different pH were tested. Of the actinobacteria tested, 88.6% showed antimicrobial activity against at least one phytopathogen, 72.1% showed a positive reaction for indoleacetic acid production, 86.8% produced siderophores and 16.2% showed a positive reaction for phosphate solubility. Isolate R18(6) was selected due to its antagonistic activity against all phytopathogenic microorganisms tested in this study. The best conditions for production were observed in the SC medium, at 30 degrees C and pH 7.0. The isolate R18(6) showed close biochemical and genetic similarity to Streptomyces pluricolorescens.
Candida albicans isoladas da cavidade bucal de crianças com síndrome de Down: ocorrência e inibição do crescimento por Streptomyces sp Candida albicans isolated from buccal cavity of children with Down's syndrome: occurrence and growth inhibition by Streptomyces sp
José Daniel Gonçalves Vieira
Full Text Available Comparação entre a presença de leveduras de Candida na cavidade bucal de crianças sem e com síndrome de Down mostrou-se estatisticamente significante no caso de crianças afetadas por esta cromossomopatia, tornando-as mais predispostas à candidíase bucal, provavelmente favorecida pelas alterações anátomo-fisiológicas da boca em decorrência da trissomia do cromossomo 21. Recidivas constantes de candidíase bucal em crianças portadoras desta alteração cromossômica levou a busca de prováveis alternativas terapêuticas. Visando determinar a atividade antifúngica de Streptomyces sp isolados de diferentes solos brasileiros, 5 cepas foram testadas frente a Candida albicans, oriundas da cavidade bucal de crianças com síndrome de Down. Observou-se que os isolados apresentaram uma diversidade de tamanho dos halos (9-31mm de diâmetro de inibição de crescimento das leveduras, sugerindo uma possível utilização em terapêutica antifúngica.Comparison of the presence of Candida yeasts in the buccal cavity of children without and with Down's syndrome showed a statistically significant difference in the case of children that were affected by this chromosomopathy, rendering them more predisposed to buccal candidiasis, probably due to anatomicophysiologic alterations of the mouth resulting from trisomy of chromosome 21. Constant recurrence of buccal candidiasis in children with this chromosomal alteration lead to the search for a possible therapeutic alternative. Seeking to determine the antifungal activity of Streptomyces sp isolated from various Brazilian soils, 5 strains have been tested for Candida albicans isolated from the buccal cavity of children with Down's syndrome. It was observed that the isolate presented a diversity in the size of the halos (9-31mm in diameter of growth inhibition of the yeasts, suggesting a possible use as a therapeutic antifungal.
Neesen, K; Volckaert, G.
Shuttle vectors for gene transfer between Streptomyces spp. and Escherichia coli have been constructed by fusion of an artificial multicopy E. coli replicon and DNA fragments of pIJ702. Stable transfer to Streptomyces lividans was obtained. Marked differences in transformation efficiency were observed when plasmid DNA isolated from E. coli GM119 was used instead of that from strain HB101.
Turton, Jane F; Shah, Jayesh; Ozongwu, Chika; Pike, Rachel
Six hundred ninety nonduplicate isolates of Acinetobacter species were identified using a combination of detection of bla(OXA-51-like) and rpoB sequence cluster analysis. Although most isolates were identified as A. baumannii (78%), significant numbers of other species, particularly A. lwoffii/genomic species 9 (8.8%), A. ursingii (4%), genomic species 3 (1.7%), and A. johnsonii (1.7%), were received, often associated with bacteremias.
Bröker, Daniel; Dietz, David; Arenskötter, Matthias; Steinbüchel, Alexander
The latex-clearing protein (Lcp(K30)) from the rubber-degrading bacterium Streptomyces sp. strain K30 is involved in the cleavage of poly(cis-1,4-isoprene), yielding isoprenoid aldehydes and ketones. Lcp homologues have so far been detected in all investigated clearing-zone-forming rubber-degrading bacteria. Internal degenerated oligonucleotides derived from lcp genes of Streptomyces sp. strain K30 (lcp(K30)), Streptomyces coelicolor strain A3(2), and Nocardia farcinica strains IFM10152 and E1 were applied in PCR to investigate whether lcp homologues occur also in the non-clearing-zone-forming rubber-utilizing bacteria Gordonia polyisoprenivorans strains VH2 and Y2K, Gordonia alkanivorans strain 44187, and Gordonia westfalica strain Kb1, which grow adhesively on rubber. The 1,230- and 1,224-bp lcp-homologous genes from G. polyisoprenivorans strain VH2 (lcp(VH2)) and G. westfalica strain Kb1 (lcp(Kb1)) were obtained after screening genomic libraries by degenerated PCR amplification, and their translational products exhibited 50 and 52% amino acid identity, respectively, to Lcp(K30). Recombinant lcp(VH2) and lcp(Kb1) harboring cells of the non-rubber-degrading Streptomyces lividans strain TK23 were able to form clearing zones and aldehydes on latex overlay-agar plates, thus indicating that lcp(VH2) and lcp(Kb1) encode functionally active proteins. Analysis by gel permeation chromatography demonstrated lower polymer concentrations and molecular weights of the remaining polyisoprenoid molecules after incubation with these recombinant S. lividans strains. Reverse transcription-PCR analysis demonstrated that lcp(VH2) was transcribed in cells of G. polyisoprenivorans strain VH2 cultivated in the presence of poly(cis-1,4-isoprene) but not in the presence of sodium acetate. Anti-Lcp(K30) immunoglobulin Gs, which were raised in this study, were rather specific for Lcp(K30) and did not cross-react with Lcp(VH2) and Lcp(Kb1). A lcp(VH2) disruption mutant was still able to grow
Arias, Anthony Argüelles; Lambert, Stéphany; Martinet, Loïc; Adam, Delphine; Tenconi, Elodie; Hayette, Marie-Pierre; Ongena, Marc; Rigali, Sébastien
Due to the necessity of iron for housekeeping functions, nutrition, morphogenesis and secondary metabolite production, siderophore piracy could be a key strategy in soil and substrate colonization by microorganisms. Here we report that mutants of bacterium Streptomyces coelicolor unable to produce desferrioxamine siderophores could recover growth when the plates were contaminated by indoor air spores of a Penicillium species and Engyodontium album. UPLC-ESI-MS analysis revealed that the HPLC fractions with the extracellular 'resuscitation' factors of the Penicillium isolate were only those that contained siderophores, i.e. Fe-dimerum acid, ferrichrome, fusarinine C and coprogen. The restored growth of the Streptomyces mutants devoid of desferrioxamine is most likely mediated through xenosiderophore uptake as the cultivability depends on the gene encoding the ABC-transporter-associated DesE siderophore-binding protein. That a filamentous fungus allows the growth of desferrioxamine non-producing Streptomyces in cocultures confirms that xenosiderophore piracy plays a vital role in nutritional interactions between these taxonomically unrelated filamentous microorganisms.
Full Text Available OBJECTIVE: To describe the antimicrobial activity of several antimicrobial agents against 97 clinical significant isolates of Enterococcus spp. MATHERIAL AND METHODS: During a 2-year prospective study at Instituto Nacional de Pediatria (National Institute of Pediatrics in Mexico City. Ninety seven strains of Enterococcus spp. (60 E. faecalis and 37 E. faecium were tested against 11 antibiotics. Susceptibility tests were performed with agar, according to the standards of the sNational Committee for Clinical Laboratory Standards (NCCLS. Isolates were screened for high-level resistance (HLR to beta-lactams, aminoglycosides, glycopeptides and other antibiotics, as well as for vancomycin-phenotypes. Differences between proportions were evaluated with chi2 of Fisher exact fest. RESULTS: Overall resistance rates to the antibiotics tested were: 17/97 (17.5% to penicillin, ampicillin, amoxicillin-clavulanate and imipenem. There was neither HLR nor beta-lactamase production; 74/97 (48.4% were resistant to erythromycin; 60% to ciprofloxacin; 31/97 (32% to gentamicin, and 55/97 (56.7% to streptomycin. Seven strains were vancomycin-resistant enterococci (VRE, all of them identified as E. faecium; 5/7 with Van A and 2/7 with Van B phenotypes. All the isolates were susceptible to linezolid. The difference in susceptibility among species was significant. CONCLUSIONS: Mutidrug-resistant enterococci is a real problem and continuous surveillance is necessary. The microbiology laboratory is the first line of defense against the spread of multiantibiotic-resistan enterococci in the hospital environment . All the strains recovered should be tested for susceptibility to ampicillin, streptomycin, gentamicin and glycopeptides.
El-Naggar, Noura El-Ahmady; Abdelwahed, Nayera A.M.; Saber, Wesam I.A.; Mohamed, Asem A.
The use of low cost agro-industrial residues for the production of industrial enzymes is one of the ways to reduce significantly production costs. Cellulase producing actinomycetes were isolated from soil and decayed agricultural wastes. Among them, a potential culture, strain NEAE-J, was selected and identified on the basis of morphological, cultural, physiological and chemotaxonomic properties, together with 16S rDNA sequence. It is proposed that strain NEAE-J should be included in the species Streptomyces albogriseolus as a representative of a novel sub-species, Streptomyces albogriseolus subsp. cellulolyticus strain NEAE-J and sequencing product was deposited in the GenBank database under accession number JN229412. This organism was tested for its ability to produce endoglucanase and release reducing sugars from agro-industrial residues as substrates. Sugarcane bagasse was the most suitable substrate for endoglucanase production. Effects of process variables, namely incubation time, temperature, initial pH and nitrogen source on production of endoglucanase by submerged fermentation using Streptomyces albogriseolus subsp. cellulolyticus have been studied. Accordingly optimum conditions have been determined. Incubation temperature of 30 °C after 6 days, pH of 6.5, 1% sugarcane bagasse as carbon source and peptone as nitrogen source were found to be the optimum for endoglucanase production. Optimization of the process parameters resulted in about 2.6 fold increase in the endoglucanase activity. Therefore, Streptomyces albogriseolus subsp. cellulolyticus coud be potential microorganism for the intended application. PMID:25242966
Noura El-Ahmady El-Naggar
Full Text Available The use of low cost agro-industrial residues for the production of industrial enzymes is one of the ways to reduce significantly production costs. Cellulase producing actinomycetes were isolated from soil and decayed agricultural wastes. Among them, a potential culture, strain NEAE-J, was selected and identified on the basis of morphological, cultural, physiological and chemotaxonomic properties, together with 16S rDNA sequence. It is proposed that strain NEAE-J should be included in the species Streptomyces albogriseolus as a representative of a novel sub-species, Streptomyces albogriseolus subsp. cellulolyticus strain NEAE-J and sequencing product was deposited in the GenBank database under accession number JN229412. This organism was tested for its ability to produce endoglucanase and release reducing sugars from agro-industrial residues as substrates. Sugarcane bagasse was the most suitable substrate for endoglucanase production. Effects of process variables, namely incubation time, temperature, initial pH and nitrogen source on production of endoglucanase by submerged fermentation using Streptomyces albogriseolus subsp. cellulolyticus have been studied. Accordingly optimum conditions have been determined. Incubation temperature of 30 ºC after 6 days, pH of 6.5, 1% sugarcane bagasse as carbon source and peptone as nitrogen source were found to be the optimum for endoglucanase production. Optimization of the process parameters resulted in about 2.6 fold increase in the endoglucanase activity. Therefore, Streptomyces albogriseolus subsp. cellulolyticus coud be potential microorganism for the intended application.
El-Naggar, Noura El-Ahmady; Abdelwahed, Nayera A M; Saber, Wesam I A; Mohamed, Asem A
The use of low cost agro-industrial residues for the production of industrial enzymes is one of the ways to reduce significantly production costs. Cellulase producing actinomycetes were isolated from soil and decayed agricultural wastes. Among them, a potential culture, strain NEAE-J, was selected and identified on the basis of morphological, cultural, physiological and chemotaxonomic properties, together with 16S rDNA sequence. It is proposed that strain NEAE-J should be included in the species Streptomyces albogriseolus as a representative of a novel sub-species, Streptomyces albogriseolus subsp. cellulolyticus strain NEAE-J and sequencing product was deposited in the GenBank database under accession number JN229412. This organism was tested for its ability to produce endoglucanase and release reducing sugars from agro-industrial residues as substrates. Sugarcane bagasse was the most suitable substrate for endoglucanase production. Effects of process variables, namely incubation time, temperature, initial pH and nitrogen source on production of endoglucanase by submerged fermentation using Streptomyces albogriseolus subsp. cellulolyticus have been studied. Accordingly optimum conditions have been determined. Incubation temperature of 30 °C after 6 days, pH of 6.5, 1% sugarcane bagasse as carbon source and peptone as nitrogen source were found to be the optimum for endoglucanase production. Optimization of the process parameters resulted in about 2.6 fold increase in the endoglucanase activity. Therefore, Streptomyces albogriseolus subsp. cellulolyticus coud be potential microorganism for the intended application.
Drautz, H; Reuschenbach, P; Zähner, H; Rohr, J; Zeeck, A
Elloramycin (1), a new antibiotic produced by Streptomyces olivaceus strain Tü 2353, was detected by chemical screening. The dark yellow compound, molecular formula C32H36O15, is weakly active against a variety of Gram-positive bacteria, especially streptomycetes and against stem cells of L-1210 leukemia. Acidic hydrolysis of the antibiotic liberated elloramycinone (3) as aglycone and 2,3,4-tri-O-methyl-L-rhamnose, which was identified as methyl glycoside 5b. The structure of elloramycin was established by comparison of the spectra (UV, 1H NMR, 13C NMR) with those of the known tetracenomycin C (2), 3 and the fact that 2 and 3 gave the same tetramethyl ether after permethylation. Elloramycin is an anthracycline-like antibiotic, the aglycone resembles tetracenomycin C, the sugar is connected in a phenolic alpha-glycosidic linkage.
Stapley, E. O.; Jackson, M.; Hernandez, S.; Zimmerman, S. B.; Currie, S. A.; Mochales, S.; Mata, J. M.; Woodruff, H. B.; Hendlin, D.
A number of actinomycetes isolated from soil were found to produce one or more members of a new family of antibiotics, the cephamycins, which are structurally related to cephalosporin C. The cephamycins were produced in submerged fermentation in a wide variety of media by one or more of eight different species of Streptomyces, including a newly described species, S. lactamdurans. These antibiotics exhibit antibacterial activity against a broad spectrum of bacteria which includes many that are resistant to the cephalosporins and penicillins. PMID:4790552
Hayes, Sabrina R; Hudon, Michael; Park, Hyun-Woo
Two novel mosquitocidal bacteria, VB17 and VB24, identified as new Bacillus species were isolated from dead mosquito larvae obtained in Florida aquatic habitats. Gas chromatographic analysis of fatty acid methyl esters (GC-FAME) and 16S rRNA sequencing indicated that VB24 is closely related to Bacillus sphaericus whereas VB17 does not have a close relationship with either Bacillus thuringiensis or B. sphaericus. Both isolates were significantly more active than B. sphaericus 2362 against Aedes taeniorhynchus, Anopheles quadrimaculatus, Culex quinquefasciatus larvae, and as active as B. sphaericus 2362 against Anopheles gambiae. Interestingly, however, both were not active against Aedes aegypti larvae, indicating some level of insecticidal specificity. Copyright © 2011 Elsevier Inc. All rights reserved.
Boudjeko, Thaddée; Tchinda, Romaric Armel Mouafo; Zitouni, Mina; Nana, Joëlle Aimée Vera Tchatchou; Lerat, Sylvain; Beaulieu, Carole
The taxonomy of an actinobacterial strain, designated JJY4(T), was established using a polyphasic approach. JJY4(T) was isolated from the rhizosphere of Chromolaena odorata in Yaoundé (Cameroon) during a project for the selection of biological control agents. Strain JJY4(T) exhibited antimicrobial activities against bacteria, fungi, and oomycetes. Strain JJY4(T) also exhibited the traits of plant growth-promoting rhizobacteria such as the solubilization of inorganic phosphate, production of siderophores and indole-3-acetic acid, and 1-aminocyclopropane-1-carboxylate deaminase activity. In planta assays performed on cocoa plantlets confirmed that strain JJY4(T) exhibited strong abilities to promote plant growth and protect against Phytophthora megakarya, the main causal agent of cocoa pod rot. The formation of rugose-ornamented spores in spiral spore chains by strain JJY4(T) is a typical feature of members found in the Streptomyces violaceusniger clade and, similar to some members of the clade, strain JJY4(T) produces geldanamycin. A phylogenetic analysis based on 16S rRNA gene sequences confirmed this classification and suggests that strain JJY4(T) be added to the subclade constituted of the type strains Streptomyces malaysiensis DSM 41697(T) and Streptomyces samsunensis DSM 42010(T). However, DNA-DNA relatedness and physiological characteristics allowed for the differentiation of strain JJY4(T) from its closest phylogenetic relatives. Based on these results, strain JJY4(T) (=NRRL B-65369, =NBRC 112705) appears to represent a novel species in the S. violaceusniger clade for which the proposed name is Streptomyces cameroonensis sp. nov.
Full Text Available Carbendazim (methyl-1H-benzimidazol-2-ylcarbamate, or MBC is a benzimidazole fungicide which is used to protect crops against the attack of fungi. MBC has a half-life of about 3-12 months and remain persistent in the environment which may lead to many harmful consequences. Besides chemical and photo-catalytic degradation of pesticides, microbial degradation has now been evolved as a much effective and safer way to eliminate these harmful compounds from the environment. However, in the literature very few reports are available where microbial community is involved in degrading MBC. Hence, the present study was planned to investigate the role of microbes isolated from the field soils for the bioremediation of MBC. Soil samples were collected from wheat fields of northern regions of India. Enrichment culture technique was employed to isolate the bacterium which was found to be growing at higher concentrations of MBC up to 500µg/ml. After biochemical and morphological analysis, the bacterium was identified as Streptomyces albogriseolus. Streptomyces albogriseolus was found to degrade MBC in a time-dependent manner from the initial concentration of 29 ppm to 285.67ppb and 62.73ppb in 24hrs and 48hrs respectively. LCMS-MS analysis was carried out to detect 2-aminobenzimidazole, a metabolite formed after degradation in 10 hrs of growth which eventually disappeared after 24hrs of growth. The strain Streptomyces albogriseolus holds a promising potential to be an efficient MBC bioremediation agent.
Irfan; Ahmad; Mir; Sudhir; Kumar; Kashyap; Sunil; Maherchandani
Objective: To study the occurrence and serotype diversity of Salmonella isolates in different species of poultry(chicken, emu and duck) and determine their resistance pattern against various antibiotics of different classes.Methods: About 507 samples comprising 202 caecal contents and 305 fecal samples from chicken, emu and duck were processed for isolation of Salmonella enterica. Salmonellae were isolated and detected by standard protocol of ISO 6579 Amendment 1: Annex D. Genetic confirmation was also made by using 16 S r RNA genus specific PCR. Serotype specific PCR was also done to detect the most common serovars viz. Salmonella Enteritidis, Salmonella Typhimurium and Salmonella Gallinarum. All obtained isolates were subjected to a set of 25 antibiotics to study their antibiogram by using Baeur-Kirby disk diffusion method.Results: Out of 507 samples processed, 32 isolates of Salmonella enterica(18 from caecal contents and 14 from faecal samples) were obtained, of which 24 belonged to 6 different serovars, 6 were untypeable and 2 were rough strains. Salmonella Enteritidis was the most predominant serotype(9), followed by Salmonella Typhimurium(5), Salmonella Virchow(4), Salmonella Gallinarum(3), Salmonella Reading(2) and Salmonella Altona(1). Antibiotic resistance pattern was maximum(100%) to oxacillin, penicillin and clindamycin, followed by ampicillin(68.75%), tetracycline(65.62%), nalidixic acid(56.25%) and colistin(46.87%). High sensitivity of isolates was recorded for chloramphenicol(96.87%) followed by meropenem(84.37%). Conclusions: Occurrence of high proportion of serovars in our study which can cause serious gastroenteritis in humans is a matter of concern. Salmonella Altona has been detected for the first time in India from poultry. This serotype is known to cause serious outbreaks of gastroenteritis in humans. Multidrug resistant isolates were recovered at high percentage which can be attributed to non-judicious use of antibiotics both in prophylaxis
Irfan Ahmad Mir; Sudhir Kumar Kashyap; Sunil Maherchandani
Objective:To study the occurrence and serotype diversity of Salmonella isolates in different species of poultry (chicken, emu and duck) and determine their resistance pattern against various antibiotics of different classes. Methods:About 507 samples comprising 202 caecal contents and 305 fecal samples from chicken, emu and duck were processed for isolation of Salmonella enterica. Salmonellae were isolated and detected by standard protocol of ISO 6579 Amendment 1:Annex D. Genetic confirmation was also made by using 16S rRNA genus specific PCR. Serotype specific PCR was also done to detect the most common serovars viz. Salmonella Enteritidis, Salmonella Typhimurium and Salmonella Gallinarum. All obtained isolates were subjected to a set of 25 antibiotics to study their antibiogram by using Baeur-Kirby disk diffusion method. Results:Out of 507 samples processed, 32 isolates of Salmonella enterica (18 from caecal contents and 14 from faecal samples) were obtained, of which 24 belonged to 6 different serovars, 6 were untypeable and 2 were rough strains. Salmonella Enteritidis was the most predominant serotype (9), followed by Salmonella Typhimurium (5), Salmonella Virchow (4), Salmonella Gallinarum (3), Salmonella Reading (2) and Salmonella Altona (1). Antibiotic resistance pattern was maximum (100%) to oxacillin, penicillin and clindamycin, followed by ampicillin (68.75%), tetracycline (65.62%), nalidixic acid (56.25%) and colistin (46.87%). High sensitivity of isolates was recorded for chloramphenicol (96.87%) followed by meropenem (84.37%). Conclusions:Occurrence of high proportion of serovars in our study which can cause serious gastroenteritis in humans is a matter of concern. Salmonella Altona has been detected for the first time in India from poultry. This serotype is known to cause serious outbreaks of gastroenteritis in humans. Multidrug resistant isolates were recovered at high percentage which can be attributed to non-judicious use of antibiotics both in
Full Text Available Penicillium is a large genus of common molds with over 400 described species; however, identification of individual species is difficult, including for those species that cause postharvest rots. In this study, blue rot fungi from stored apples and pears were isolated from a variety of hosts, locations, and years. Based on morphological and cultural characteristics and partial amplification of the β-tubulin locus, the isolates were provisionally identified as several different species of Penicillium. These isolates were investigated further using a suite of molecular DNA markers and compared to sequences of the ex-type for cognate species in GenBank, and were identified as P. expansum (3 isolates, P. solitum (3 isolates, P. carneum (1 isolate, and P. paneum (1 isolate. Three of the markers we used (ITS, internal transcribed spacer rDNA sequence; benA, β-tubulin; CaM, calmodulin were suitable for distinguishing most of our isolates from one another at the species level. In contrast, we were unable to amplify RPB2 sequences from four of the isolates. Comparison of our sequences with cognate sequences in GenBank from isolates with the same species names did not always give coherent data, reinforcing earlier studies that have shown large intraspecific variability in many Penicillium species, as well as possible errors in some sequence data deposited in GenBank.
Said Mohamed Daboor
Full Text Available Objective: To find the best actinomycete that has potential application value in the heavy metal remediation due to its special morphological and physiological metabolism. Methods: In some areas of River Nile, Egypt, a total of 67 actinomycete isolates (17 isolates from surface water and 50 from sediment were identified. In addition, the studied area was characterized by a large amount of submerged macrophyte species Ceratophyllum demersum, one free floating species Eichhornia crassipes and two emergent species Polygonum tomentosum and Saccharum spontaneum with the highest biomass production values. Many methods are used in this research like qualitative evaluation of heavy metals, minimum inhibitory concentration of heavy metal determination, metal binding assay, heavy metal assessment, etc. Results: Many actinomycetes isolates were isolated from River Nile, Egypt, the absorbent efficiency of one isolate Streptomyces chromofuscusK101 showed the most efficient metal binding activity. The adsorption process of Zn2+ , Pb2+ and Fe 2+ single or mixture metal ions was investigated, where the order of adsorption potential ( Zn2+ >Pb2+ >Fe 2+ was observed in single metal reaction. The adsorption in mixed metal reactions was the same order as in single-metal ion with a significant decrease in Fe 2+ and Pb2+ adsorption. Conclusions: In conclusion the metal adsorption reactions were very fast, pH dependent and culture age-independent, suggestive of a physicochemical reaction between cell wall components and heavy metal ions. The absorbent removal efficiency was determined as a function of ion concentration, pH and temperature.
Kitani, Shigeru; Miyamoto, Kiyoko T; Takamatsu, Satoshi; Herawati, Elisa; Iguchi, Hiroyuki; Nishitomi, Kouhei; Uchida, Miho; Nagamitsu, Tohru; Omura, Satoshi; Ikeda, Haruo; Nihira, Takuya
Gram-positive bacteria of the genus Streptomyces are industrially important microorganisms, producing >70% of commercially important antibiotics. The production of these compounds is often regulated by low-molecular-weight bacterial hormones called autoregulators. Although 60% of Streptomyces strains may use γ-butyrolactone-type molecules as autoregulators and some use furan-type molecules, little is known about the signaling molecules used to regulate antibiotic production in many other members of this genus. Here, we purified a signaling molecule (avenolide) from Streptomyces avermitilis--the producer of the important anthelmintic agent avermectin with annual world sales of $850 million--and determined its structure, including stereochemistry, by spectroscopic analysis and chemical synthesis as (4S,10R)-10-hydroxy-10-methyl-9-oxo-dodec-2-en-1,4-olide, a class of Streptomyces autoregulator. Avenolide is essential for eliciting avermectin production and is effective at nanomolar concentrations with a minimum effective concentration of 4 nM. The aco gene of S. avermitilis, which encodes an acyl-CoA oxidase, is required for avenolide biosynthesis, and homologs are also present in Streptomyces fradiae, Streptomyces ghanaensis, and Streptomyces griseoauranticus, suggesting that butenolide-type autoregulators may represent a widespread and another class of Streptomyces autoregulator involved in regulating antibiotic production.
Three new aflatoxin-producing species belonging to Aspergillus section Flavi are described, Aspergillus mottae, Aspergillus sergii and Aspergillus transmontanensis. These species were isolated from Portuguese almonds and maize. An investigation examining morphology, extrolites and molecular data was...
Mani, A.; Anil, A.C.; Leliaert, F.; Delany, J.; Mesbahi, E.
A new species of Tetraselmis, T. indica Arora & Anil, was isolated from nanoplankton collected from salt pans in Goa (India) and is described based on morphological, ultrastructural, 18S rRNA gene sequence and genome size data. The species...
Kishii, Kozue; Kikuchi, Ken; Yoshida, Atsushi; Okuzumi, Katsuko; Uetera, Yushi; Yasuhara, Hiroshi; Moriya, Kyoji
Carbapenem-resistant Acinetobacter baumannii has rapidly spread worldwide. This study investigated antibiotic susceptibility and genotypic resistance of 123 consecutive blood culture isolates of Acinetobacter species collected between 2003 and 2011 in two Japanese hospitals. The isolates were assigned to 13 species. Carbapenem resistance was detected in four isolates. Only one A. baumannii isolate had blaOXA-23 together with ISAba1; the remaining three isolates had IMP-1 metallo-β-lactamase. Quinolone resistance was detected in five isolates that had point mutations in the quinolone resistance-determining region. The predominance of various non-A. baumannii species and low prevalence of carbapenem resistance among blood culture isolates of Acinetobacter species in two Japanese hospitals were confirmed.
Matsushima, P; Baltz, R H
We explored different methods of introducing DNA into 'Streptomyces toyocaensis' and Streptomyces virginiae to construct stable recombinant strains. Plasmid pIJ702 isolated from Streptomyces lividans transformed protoplasts of 'S. toyocaensis' at a frequency of 7 x 10(3) transformants (mu g DNA)-1. pIJ702 prepared from 'S. toyocaensis' transformed 'S. toyocaensis' protoplasts at a frequency of 1 center dot 5 x 10(5) (mu g DNA)-1, suggesting that 'S. toyocaensis' expresses restriction and modification. Plasmid pRHB126 was transduced by bacteriophage FP43 into 'S. toyocaensis' at a frequency of 1.2 x 10(-6) (p.f.u)-1. Plasmids pOJ436 and pRHB304 were introduced into 'S. toyocaensis' by conjugation from Escherichia coli S17-1 at frequencies of about 2 x 10(-4) and 1 x 10(-4) per recipient, respectively. Analysis of several exconjugants indicated that pOJ436 and pRHB304 inserted into a unique phiC31 attB site and that some of the insertions had minimal deleterious effects on glycopeptide A47934 production. The results indicate that 'S. toyocaensis' is a suitable host for gene cloning, whereas S. virginiae does not appear to be.
Kim, Ki Hyun; Ramadhar, Timothy R.; Beemelmanns, Christine
We report a preliminary functional and complete structural characterization of a highly unusual geldanamycin analog, natalamycin A, that was isolated from Streptomyces strain M56 recovered from a South African nest of Macrotermes natalensis termites. Bioassay-guided fractionation based on antifun......We report a preliminary functional and complete structural characterization of a highly unusual geldanamycin analog, natalamycin A, that was isolated from Streptomyces strain M56 recovered from a South African nest of Macrotermes natalensis termites. Bioassay-guided fractionation based...
García Bernal, M; Trabal Fernández, N; Saucedo Lastra, P E; Medina Marrero, R; Mazón-Suástegui, J M
To determine the composition and diversity of the microbiota associated to Crassostrea sikamea treated during 30 days with Streptomyces strains N7 and RL8. DNA was extracted from oysters followed by 16S rRNA gene amplification and pyrosequencing. The highest and lowest species diversity richness was observed in the initial and final control group, whereas Streptomyces-treated oysters exhibited intermediate values. Proteobacteria was the most abundant phylum (81·4-95·1%), followed by Bacteroidetes, Actinobacteria and Firmicutes. The genera Anderseniella, Oceanicola, Roseovarius, Ruegeria, Sulfitobacter, Granulosicoccus and Marinicella encompassed the core microbiota of all experimental groups. The genus Bacteriovorax was detected in all groups except in the final control and the depurated N7, whereas Vibrio remained undetected in all Streptomyces-treated groups. RL8 was the only group that harboured the genus Streptomyces in its microbiota. Principal component analysis showed that Streptomyces strains significantly changed oyster microbiota with respect to the initial and final control. Crassostrea sikamea treated with Streptomyces showed high species diversity and a microbiota composition shift, characterized by keeping the predator genus Bacteriovorax and decreasing the pathogenic Vibrio. This is the first culture-independent study showing the effect of Streptomyces over the oyster microbiota. It also sheds light about the potential use of Streptomyces to improve mollusc health and safety for consumers after the depuration process. © 2016 The Society for Applied Microbiology.
Rangarajan, M; Ravindran, A D; Hariharan, K
A lysogenic Streptomyces sp., strain NS.A4, which was isolated from the nodule surface of black gram (Vigna mungo (L.) Hepper), was found to inhibit rhizobia of fast-and slow-growing strains of cowpeas and soybeans. It exhibited plaques when there was a change in cultural conditions. Repeated culturing of the organism in nutrient agar and broth confirmed the infection of Streptomyces sp. strain NS.A4 by an actinophage. Addition of the culture filtrate of Streptomyces sp. strain NS.A4 to shaken broth cultures of three other Streptomyces spp. resulted in phage infection.
Bi, Shu-Feng; Guo, Zhi-Kai; Jiang, Nan; Jiao, Rui-Hua; Ge, Hui-Ming; Tan, Ren-Xiang
A new alkaloid was isolated from the ethyl acetate extract of the culture of a termite-associated Streptomyces koyangensis BY-4. The structure of 1 was elucidated by using MS, NMR, electronic circular dichroism data, and computational approaches. Compound 1 showed weak antimicrobial activities against a panel of test microbes.
Xiu Chao XIE; Wen Li MEI; You Xing ZHAO; Kui HONG; Hao Fu DAI
A new degraded sesquiterpene was isolated from the marine actinomycete Streptomyces sp. 0616208. Its structure was elucidated as (1α, 4aα, 5α, 7β, 8aβ)-5, 8a-dimethyl-decahydrona-phthalene-1, 4a, 7-triol on the basis of spectroscopic data.
Full Text Available We propose a new species, Christensenella timonensis, strain Marseille-P2437T (CSUR P2437T, which was isolated from gut microbiota of a 66-year-old patient as a part of culturomics study. C. timonensis represents the second species isolated within the Christensenella genus.
Joseph, Noyal Mariya; Harish, Belgode Narasimha; Sistla, Sujatha; Thappa, Devinder Mohan; Parija, Subhash Chandra
A 29-year-old woman presented with multiple painful swelling with discharging sinuses over the scalp. Histopathological examination of the biopsy tissue was suggestive of actinomycotic mycetoma. Streptomyces spp. was isolated from blood culture. The patient was successfully treated with trimethoprim-sulfamethoxazole and crystalline penicillin. This case is reported because of the rare occurrence of bacteremia by Streptomyces spp. secondary to subcutaneous actinomycotic mycetoma. Moreover, an interesting association between successive two pregnancies and occurrence of mycetoma of the scalp was observed in this case.
Eddouzi, J.; Hofstetter, V.; Groenewald, M.; Manai, M.; Sanglard, S.
From a collection of yeast isolates isolated from patients in Tunisian hospitals between September 2006 and July 2010, the yeast strain JEY63 (CBS 12513), isolated from a 50-year-old male that suffered from oral thrush, could not be identified to the species level using conventional methods used in
A polyphasic study was carried out to establish the taxonomic status of an Atacama Desert isolate, Streptomyces strain C34T, which synthesises novel antibiotics, the chaxalactins and chaxamycins. The organism was shown to have chemotaxonomic, cultural, and morphological properties consistent with it...
Full Text Available Objectives: Demand for new antibiotic is rising up due to continuous resistance risk against conventional antibiotic.This attempt was taken to find out a novel antimicrobial metabolite.Methods: Chili field antagonistic actinomycetes Streptomyces albolongus was isolated and tested for optimum antimicrobialmetabolite production. Primary screening was done by selective media and antibiotic assay was done by agarcup plate method. Fermented product was recovered by separating funnel using suitable solvent.Results: Maximum antimicrobial metabolite production was found at temperature 35°C and pH 9.0 and on 6th day ofincubation. The medium consisting of corn steep liquor (0.2%, glucose (1.0%, NaCl (0.5%, K2HPO4 (0.1% was screenedout as suitable medium for maximum antimicrobial production. Sucrose was found as the best carbon source amongfour sources. The antimicrobial metabolite was found to be stable at pH and temperature up to 11.0 and 100°C respectively.The active agent was best extracted with chloroform. The antimicrobial spectrum of the metabolite was wideand shows activity against Shigella dysenteriae (AE14612, Shigella sonnei (CRL, ICDDR, B, Salmonella typhi (AE14296,Vibrio cholerae (AE14748, Pseudomonas aeruginosa (CRL, ICDDR, B, Bacillus cereus (BTCC19, Staphylococcus aureus(ATCC6538, Bacillus subtilis (BTTC17 and Bacillus megaterium (BTTC18.Conclusions: The findings of antibacterial activity of S. albolongus against several species of human pathogens includingboth Gram-positive and Gram-negative bacteria indicated that our produced material might be an alternative antimicrobialsubstance to control human diseases. J Microbiol Infect Dis 2013; 3(2: 75-82Key words: Streptomyces albolongus, antimicrobial metabolite, optimum production, antimicrobial spectrum
Bajić Bojana Ž.
Full Text Available Numerous species of the genus Streptomyces, on the appropriate cultivation medium in the process of submerged biosynthesis, as a product of the secondary metabolism, and under aerobic conditions synthesize pharmacologically active compounds. The aim of presented study was optimization of different nitrogen sources in the cultivation medium for the production of antimycotics using a strain of Streptomyces spp. isolated from the environment. Experiments were carried out in accordance with Box-Behnken design with three factors at three levels (peptone: 3.0 g/l, 7.0 g/l and 11.0 g/l; yeast extract: 1.0 g/l, 3.0 g/l and 5.0 g/l; soybean meal: 5.0 g/l, 15.0 g/l and 25.0 g/l and three repetitions in the central point. Cultivation mediums were analyzed for determination of residual sugar, residual nitrogen, pellet diameter and RNA. Also, antimycotic activity of the obtained cultivation mediums was determined using diffusion disc method on the Aspergillus spp. as the test microorganism. For the optimization of selected parameters, a Response Surface Methodology was used and the obtained data were analyzed using the software package DESIGN EXPERT 8.1. Achieved model with a coefficient of determination (R of 0.952 predicted that the maximum inhibition zone diameter (24.0 mm against microorganism Aspergillus spp. and the minimum amount of residual sugar (0.551528 g/l under applied experimental conditions was produced when the contents of varied nitrogen sources were: peptone 11.0 g/l, yeast extract 4.32 g/l and soybean meal 25.00 g/l.
Siitonen, Vilja; Räty, Kaj; Metsä-Ketelä, Mikko
The "Streptomyces genetics and secondary metabolism" laboratory course gives an introduction to the versatile soil dwelling Gram-positive bacteria Streptomyces and their secondary metabolism. The course combines genetic modification of Streptomyces; growing of the strain and protoplast preparation, plasmid isolation by alkaline lysis and phenol precipitation, digestions, and ligations prior to protoplast transformation, as well as investigating the secondary metabolites produced by the strains. Thus, the course is a combination of microbiology, molecular biology, and chemistry. After the course the students should understand the relationship between genes, proteins, and the produced metabolites. © 2016 by The International Union of Biochemistry and Molecular Biology, 44(5):492-499, 2016. © 2016 The International Union of Biochemistry and Molecular Biology.
Heras-Cañas, Victor; Ros, Luis; Sorlózano, Antonio; Gutiérrez-Soto, Blanca; Navarro-Marí, José María; Gutiérrez-Fernández, José
Candiduria detection in hospitalized or immunocompromised patients is of great clinical significance. The aim of our study was to describe the isolation frequency of significant species of yeasts in urine samples processed in our hospital during the period 2010- 2013, and to analyze their susceptibility to commonly used antifungal agents. Species identification was performed by seeding on a chromogenic medium, the filamentation test and automated systems (ASM Vitek and MALDI Biotyper), while susceptibility was determined using the ASM Vitek system. Of the 632 yeast isolates in urine, 371 were Candida albicans species and 261 non-C. albicans Candida spp. The species with the highest number of resistant isolates were Candida glabrata and Candida krusei. Based on the results obtained, we believe that species identification and the susceptibility study should be current practice in the laboratories when species other than C. albicans are isolated.
Small signalling molecules called γ-butyrolactones are mainly produced by Streptomyces species in which they regulate antibiotic production and morphological differentiation. Their molecular mechanism of action has recently been unravelled in several streptomycetes, revealing a diverse and complex s
Chai, Yen Yen; Kahar, Ummirul Mukminin; Md Salleh, Madihah; Md Illias, Rosli; Goh, Kian Mau
Two thermophilic bacteria (SK3-4 and DT3-1) were isolated from the Sungai Klah (SK) and Dusun Tua (DT) hot springs in Malaysia. The cells from both strains were rod-shaped, stained Gram positive and formed endospores. The optimal growth of both strains was observed at 55 degrees C and pH 7. Strain DT3-1 exhibited a higher tolerance to chloramphenicol (100 microg ml(-1)) but showed a lower tolerance to sodium chloride (2%, w/v) compared to strain SK3-4. Phylogenetic analysis based on 16S rRNA gene sequences revealed that both strains belong to the genus Anoxybacillus. High concentrations of 15:0 iso in the fatty acid profiles support the conclusion that both strains belong to the genus Anoxybacillus and exhibit unique fatty acid compositions and percentages compared to other Anoxybacillus species. The DNA G + C contents were 42.0 mol% and 41.8 mol% for strains SK3-4 and DT3-1, respectively. Strains SK3-4 and DT3-1 were able to degrade pullulan and to produce maltotriose and glucose, respectively, as their main end products. Based on phenotypic and chemotaxonomic characteristics, 16S rRNA gene sequences, and the DNA G + C content, we propose that strains SK3-4 and DT3-1 are new pullulan-degrading Anoxybacillus strains.
Murad Nur Baiti Abd
Full Text Available Fruit rot of tomato is a serious disease caused by Fusarium species. Sampling was conducted throughout Selangor, Malaysia and fungal species identification was conducted based on morphological and gene encoding translation elongation factor 1-α (tef1-α sequence analysis. Five species of Fusarium were discovered namely F. oxysporum (including F. oxysporum f. sp. lycopersici, F. solani, F. equiseti, F. proliferatum and F. verticillioides. Our results provide additional information regarding the diversity of Fusarium species associated with fruit rot disease of tomato.
Fernandes, E.K.K.; Moraes, A.M.L.; Pacheco, R.S.; Rangel, D.E.N.; Miller, M.P.; Bittencourt, V.R.E.P.; Roberts, D.W.
Aims: The genetic diversity of Beauveria bassiana was investigated by comparing isolates of this species to each other (49 from different geographical regions of Brazil and 4 from USA) and to other Beauveria spp. Methods and Results: The isolates were examined by multilocus enzyme electrophoresis (MLEE), amplified fragment length polymorphism (AFLP), and rDNA sequencing. MLEE and AFLP revealed considerable genetic variability among B. bassiana isolates. Several isolates from South and Southeast Brazil had high similarity coefficients, providing evidence of at least one population with clonal structure. There were clear genomic differences between most Brazilian and USA B. bassiana isolates. A Mantel test using data generated by AFLP provided evidence that greater geographical distances were associated with higher genetic distances. AFLP and rDNA sequencing demonstrated notable genotypic variation between B. bassiana and other Beauveria spp. Conclusion: Geographical distance between populations apparently is an important factor influencing genotypic variability among B. bassiana populations in Brazil. Significance and Impact of the Study: This study characterized many B. bassiana isolates. The results indicate that certain Brazilian isolates are considerably different from others and possibly should be regarded as separate species from B. bassiana sensu latu. The information on genetic variation among the Brazilian isolates, therefore, will be important to comprehending the population structure of B. bassiana in Brazil. ?? 2009 The Society for Applied Microbiology.
tions like HIV/AIDS.13 On the other hand, HIV negative individuals may ... didiasis in HIV patients, the relationship between faecal .... while 40 (65.6%) were non-albicans species. ... dominant species in patients that were not on ARV. Other.
Castro-Melchor, Marlene; Charaniya, Salim; Karypis, George; Takano, Eriko; Hu, Wei-Shou
Background: The onset of antibiotics production in Streptomyces species is co-ordinated with differentiation events. An understanding of the genetic circuits that regulate these coupled biological phenomena is essential to discover and engineer the pharmacologically important natural products made b
The bacterial pathogen Streptomyces scabies produces lesions on potato tubers, reducing their marketability and profitability. M6 and 524-8 are two closely related inbred diploid lines of the wild potato species Solanum chacoense. After testing in both field and greenhouse assays, it was found that ...
Full Text Available Fusarium proliferatum has been reported on garlic in the Northwest USA, Spain and Serbia, causing water-soaked tan-colored lesions on cloves. In this work, Fusarium proliferatum was isolated from 300 symptomatic garlic bulbs. Morphological identification of Fusarium was confirmed using species-specific PCR assays and EF-1α sequencing. Confirmation of pathogenicity was conducted with eighteen isolates. Six randomly selected F. proliferatum isolates from garlic were tested for specific pathogenicity and screened for fusaric acid production. Additionally, pathogenicity of each F. proliferatum isolate was tested on healthy seedlings of onion (Allium cepa, leek (A. porrum, scallions (A. fistulosum, chives (A. schoenoprasum and garlic (A. sativum. A disease severity index (DSI was calculated as the mean severity on three plants of each species with four test replicates. Symptoms on onion and garlic plants were observed three weeks after inoculation. All isolates tested produced symptoms on all varieties inoculated. Inoculation of F. proliferatum isolates from diseased garlic onto other Allium species provided new information on host range and pathogenicity. The results demonstrated differences in susceptibility with respect to host species and cultivar. The F. proliferatum isolates tested all produced fusaric acid (FA; correlations between FA production and isolate pathogenicity are discussed. Additionally, all isolates showed the presence of the FUM1 gene suggesting the ability of Spanish isolates to produce fumonisins.
Full Text Available Fusarium head blight (FHB caused by the filamentous fungus Fusarium graminearum is one of the most severe diseases threatening the production of small grains. Infected grains are often contaminated with mycotoxins such as zearalenone and trichothecences. During survey of contamination by FHB in rice grains, we found a bacterial isolate, designated as BN1, antagonistic to F. graminearum. The strain BN1 had branching vegetative hyphae and spores, and its aerial hyphae often had long, straight filaments bearing spores. The 16S rRNA gene of BN1 had 100% sequence identity with those found in several Streptomyces species. Phylogenetic analysis of ITS regions showed that BN1 grouped with S. sampsonii with 77% bootstrap value, suggesting that BN1 was not a known Streptomyces species. In addition, the efficacy of the BN1 strain against F. graminearum strains was tested both in vitro and in vivo. Wheat seedling length was significantly decreased by F. graminearum infection. However, this effect was mitigated when wheat seeds were treated with BN1 spore suspension prior to F. graminearum infection. BN1 also significantly decreased FHB severity when it was sprayed onto wheat heads, whereas BN1 was not effective when wheat heads were point inoculated. These results suggest that spraying of BN1 spores onto wheat heads during the wheat flowering season can be efficient for plant protection. Mechanistic studies on the antagonistic effect of BN1 against F. graminearum remain to be analyzed.
Scatamburlo, T M; Yamazi, A K; Cavicchioli, V Q; Pieri, F A; Nero, L A
Pseudomonas spp. are usually associated with spoilage microflora of dairy products due to their proteolytic potential. This is of particular concern for protein-based products, such as goat milk cheeses and fermented milks. Therefore, the goal of the present study was to characterize the proteolytic activity of Pseudomonas spp. isolated from goat milk. Goat milk samples (n=61) were obtained directly from bulk tanks on dairy goat farms (n=12), and subjected to a modified International Organization for Standardization (ISO) protocol to determine the number and proteolytic activity of Pseudomonas spp. Isolates (n=82) were obtained, identified by PCR, and subjected to pulsed-field gel electrophoresis with XbaI macro-restriction. Then, the isolates were subjected to PCR to detect the alkaline protease gene (apr), and phenotypic tests were performed to check proteolytic activity at 7°C, 25°C, and 35°C. Mean Pseudomonas spp. counts ranged from 2.9 to 4.8 log cfu/mL, and proteolytic Pseudomonas spp. counts ranged from 1.9 to 4.6 log cfu/mL. All isolates were confirmed to be Pseudomonas spp., and 41 were identified as Pseudomonas fluorescens, which clustered into 5 groups sharing approximately 82% similarity. Thirty-six isolates (46.9%) were positive for the apr gene; and 57 (69.5%) isolates presented proteolytic activity at 7°C, 82 (100%) at 25°C, and 64 (78%) at 35°C. The isolates were distributed ubiquitously in the goat farms, and no relationship among isolates was observed when the goat farms, presence of apr, pulsotypes, and proteolytic activity were taken into account. We demonstrated proteolytic activity of Pseudomonas spp. present in goat milk by phenotypic and genotypic tests and indicated their spoilage potential at distinct temperatures. Based on these findings and the ubiquity of Pseudomonas spp. in goat farm environments, proper monitoring and control of Pseudomonas spp. during production are critical.
Full Text Available Streptomyces sp was conducted from December 2010 - June 2011 at the Laboratoryof Microbiology, Biology Department, Math and Science Faculty, UdayanaUniversity Bukit Jimbaran-Bali. Implementation stages of the research consisted ofisolation and testing of the antibiotic activity Streptomyces sp to inhibit growthbacterial pathogens Erwinia sp as a cause of disease in plants fallen foul (Soft rot ofAloe barbadensis Mill.The results of this study have eight isolates of Streptomyces spwith macroscopic and microscopic characters are varied. Furthermore, all isolateswere obtained and then tested against antibiotic activity to inhibit growth the bacteriaErwinia sp. Test results obtained by Streptomyces sp that has the most effective ininhibiting the ability of the bacteria Erwinia sp isolates are Streptomyces sp2for (45%.
Auffret, Marc; Pilote, Alexandre; Proulx, Emilie; Proulx, Daniel; Vandenberg, Grant; Villemur, Richard
Geosmin and 2-methylisoborneol (MIB) have been associated with off-flavour problems in fish and seafood products, generating a strong negative impact for aquaculture industries. Although most of the producers of geosmin and MIB have been identified as Streptomyces species or cyanobacteria, Streptomyces spp. are thought to be responsible for the synthesis of these compounds in indoor recirculating aquaculture systems (RAS). The detection of genes involved in the synthesis of geosmin and MIB can be a relevant indicator of the beginning of off-flavour events in RAS. Here, we report a real-time polymerase chain reaction (qPCR) protocol targeting geoA sequences that encode a germacradienol synthase involved in geosmin synthesis. New geoA-related sequences were retrieved from eleven geosmin-producing Actinomycete strains, among them two Streptomyces strains isolated from two RAS. Combined with geoA-related sequences available in gene databases, we designed primers and standards suitable for qPCR assays targeting mainly Streptomyces geoA. Using our qPCR protocol, we succeeded in measuring the level of geoA copies in sand filter and biofilters in two RAS. This study is the first to apply qPCR assays to detect and quantify the geosmin synthesis gene (geoA) in RAS. Quantification of geoA in RAS could permit the monitoring of the level of geosmin producers prior to the occurrence of geosmin production. This information will be most valuable for fish producers to manage further development of off-flavour events.
Dec 17, 2015 ... Aspergillus species) in feeds used in poultry farms in Sokoto metropolis. ... potential exists for the production of mycotoxins that may be of both veterinary public health ... mycoflora can negatively affect feed quality; these.
Full Text Available The exploration of the human microbiome was recently revolutionized by microbial culturomics and taxonogenomics. Thanks to this approach, we report here the main characteristics of Romboutsia timonensis strain Marseille-P326, a new bacterium isolated from the right human colon by colonoscopy in a 63-year-old French man with severe anaemia with melaena.
Illnait-Zaragozí, M T; Martínez-Machín, G F; Fernández-Andreu, C M; Perurena-Lancha, M R; Theelen, B; Boekhout, T; Meis, J F; Klaassen, C H
Cryptococcus isolates from Cuban patients were identified as C. neoformans var. grubii. Although this species has since long been associated with bird droppings, a recent genotyping study provided strong evidence for additional origins of exposure. We sampled different species of trees in Havana, Cuba to identify other potential sources of exposure to this fungus. A total of 662 samples were collected from 331 trees and cacti from Havana, Cuba. Initial selection of the isolates was carried out by conventional techniques. Isolates were further characterised using a combination of AFLP analysis and DNA sequence analysis. Identification by conventional methods yielded 121 C. neoformans and 61 C. gattii isolates. Molecular analyses showed that none of these isolates was C. gattii and only one isolate proved to be C. neoformans var. grubii. A total of 27 different other species were identified. The most prevalent species was C. heveanensis (33%). Sixty-five unidentifiable isolates segregated into ten potentially novel species. Conventional cultivation methods have a low specificity for C. neoformans complex and molecular analyses need to be applied to confirm identification of isolates from environmental sources. Environmental niches responsible for most of human cryptococcal infections in Cuba remain to be identified. © 2012 Blackwell Verlag GmbH.
Prigent, Gwénolé; Aït-Ammar, Nawel; Levesque, Eric; Fekkar, Arnaud; Costa, Jean-Marc; El Anbassi, Sarra; Foulet, Françoise; Duvoux, Christophe; Merle, Jean-Claude; Dannaoui, Eric; Botterel, Françoise
Liver transplant recipients are at risk of invasive fungal infections, especially candidiasis. Echinocandin is recommended as prophylactic treatment but is increasingly associated with resistance. Our aim was to assess echinocandin drug resistance in Candida spp. isolated from liver transplant recipients treated with this antifungal class. For this, all liver-transplanted patients in a University Hospital (Créteil, France) between January and June of 2013 and 2015 were included. Susceptibilities of Candida isolates to echinocandins were tested by Etest and the EUCAST reference method. Isolates were analyzed by FKS sequencing and genotyped based on microsatellites or multilocus sequence typing (MLST) profiles. Ninety-four patients were included, and 39 patients were colonized or infected and treated with echinocandin. Echinocandin resistance appeared in 3 (8%) of the treated patients within 1 month of treatment. One patient was colonized by resistant Candida glabrata, one by resistant Candida dubliniensis, and one by resistant Candida albicans Molecular analysis found three mutations in FKS2 HS1 (F659S, S663A, and D666E) for C. glabrata and one mutation in FKS1 HS1 (S645P) for C. dubliniensis and C. albicans Susceptible and resistant isolates belonged to the same genotype. To our knowledge, this is the first study on echinocandin resistance in Candida spp. in a liver transplant population. Most resistant isolates were found around/in digestive sites, perhaps due to lower diffusion of echinocandin in these sites. This work documents the risk of emergence of resistance to echinocandin, even after short-term treatment. Copyright © 2017 Prigent et al.
任迁琪; 宋星林; 吴洪生; 李季; 陈素云; 徐亚; 姚东良; 赵月; 陈健; 李美颖
旨在筛选出能高效抑制西瓜枯萎病病原菌尖孢镰刀菌（Fusarium oxysporum f. sp. niveum）的天然拮抗菌。采用梯度稀释法分离连作辣椒根际土中的放线菌，纯化后用平板对峙法筛选出有拮抗作用的菌株，采用盆钵育苗基质拌菌法接种，进行盆栽抗病试验，并对拮抗效果最佳的菌株进行鉴定。结果显示，从连作辣椒根际土中，分离筛选出10株对西瓜枯萎病有拮抗作用的放线菌，其中菌株 F22、F23、F32的拮抗作用显著。96 h 抑菌圈直径分别达到1.70、1.17和1.47 cm。盆栽试验结果表明，拮抗菌株 F32对西瓜枯萎病30 d 的防治效果最佳，防效率达到66.7%，且抗菌谱性广。根据表型特征、生理生化特性和16S rDNA 分子鉴定结果可知，菌株 F32为链霉菌属（Streptomyces sp.），且对西瓜枯萎病有良好的防治作用，是一株具有应用价值的生防菌株。%The objective is to select a natural antagonistic bacterium efficiently controlling Fusarium oxysporum f. sp. niveum that is the pathogenic bacterium of the fusarium wilt of watermelon. Using the method of gradient dilution to isolate actinomycetes from rhizosphere soil of continuous cropping pepper,it was purified and the plate confrontation method was applied to select an antagonistic strain. Then the method of basin nursery substrates mixed with bacteria was for inoculation,a potted plant disease-resistant experiment was carried out,and the strains with the optimal antagonistic effects were identified. The results showed that 10 strains of actinomycetes with the antagonistic effects to fusarium wilt of watermelon were isolated from the growing pepper rhizosphere soil,and 3 of them(F22,F23 and F32)presented the significantly antagonistic effects. After 96 hours the diameter of the inhibition zone was 1.70 cm,1.17 cm and 1.47 cm respectively. The result of the potted plant experiments demonstrated that the control effect of the
Munguía-Rosas, Miguel A; Montiel, Salvador
Studies of the effects of patch size and isolation on plant species density have yielded contrasting results. However, much of the available evidence comes from relatively recent anthropogenic forest fragments which have not reached equilibrium between extinction and immigration. This is a critical issue because the theory clearly states that only when equilibrium has been reached can the number of species be accurately predicted by habitat size and isolation. Therefore, species density could be better predicted by patch size and isolation in an ecosystem that has been fragmented for a very long time. We tested whether patch area, isolation and other spatial variables explain variation among forest patches in plant species density in an ecosystem where the forest has been naturally fragmented for long periods of time on a geological scale. Our main predictions were that plant species density will be positively correlated with patch size, and negatively correlated with isolation (distance to the nearest patch, connectivity, and distance to the continuous forest). We surveyed the vascular flora (except lianas and epiphytes) of 19 forest patches using five belt transects (50×4 m each) per patch (area sampled per patch = 0.1 ha). As predicted, plant species density was positively associated (logarithmically) with patch size and negatively associated (linearly) with patch isolation (distance to the nearest patch). Other spatial variables such as patch elevation and perimeter, did not explain among-patch variability in plant species density. The power of patch area and isolation as predictors of plant species density was moderate (together they explain 43% of the variation), however, a larger sample size may improve the explanatory power of these variables. Patch size and isolation may be suitable predictors of long-term plant species density in terrestrial ecosystems that are naturally and anthropogenically fragmented.
Miguel A Munguía-Rosas
Full Text Available Studies of the effects of patch size and isolation on plant species density have yielded contrasting results. However, much of the available evidence comes from relatively recent anthropogenic forest fragments which have not reached equilibrium between extinction and immigration. This is a critical issue because the theory clearly states that only when equilibrium has been reached can the number of species be accurately predicted by habitat size and isolation. Therefore, species density could be better predicted by patch size and isolation in an ecosystem that has been fragmented for a very long time. We tested whether patch area, isolation and other spatial variables explain variation among forest patches in plant species density in an ecosystem where the forest has been naturally fragmented for long periods of time on a geological scale. Our main predictions were that plant species density will be positively correlated with patch size, and negatively correlated with isolation (distance to the nearest patch, connectivity, and distance to the continuous forest. We surveyed the vascular flora (except lianas and epiphytes of 19 forest patches using five belt transects (50×4 m each per patch (area sampled per patch = 0.1 ha. As predicted, plant species density was positively associated (logarithmically with patch size and negatively associated (linearly with patch isolation (distance to the nearest patch. Other spatial variables such as patch elevation and perimeter, did not explain among-patch variability in plant species density. The power of patch area and isolation as predictors of plant species density was moderate (together they explain 43% of the variation, however, a larger sample size may improve the explanatory power of these variables. Patch size and isolation may be suitable predictors of long-term plant species density in terrestrial ecosystems that are naturally and anthropogenically fragmented.
Sørensen, Annette; Lübeck, Peter S.; Lübeck, Mette
A novel species, Aspergillus saccharolyticus sp. nov., belonging to the Aspergillus section Nigri group is described. This species was isolated in Denmark from treated hardwood. Its taxonomic status was determined using a polyphasic taxonomic approach including phenotypic (morphology and extrolite...... Aspergillus species that is morphologically similar to Aspergillus japonicus and Aspergillus aculeatus, but has a totally different extrolite profile compared to any known Aspergillus species. The type strain of A. saccharolyticus sp. nov. is CBS 127449T ( = IBT 28509T)....
Sørensen, Annette; Lübeck, Peter S.; Lübeck, Mette
A novel species, Aspergillus saccharolyticus sp. nov., belonging to the Aspergillus section Nigri group is described. This species was isolated in Denmark from treated hardwood. Its taxonomic status was determined using a polyphasic taxonomic approach including phenotypic (morphology and extrolite...... Aspergillus species that is morphologically similar to Aspergillus japonicus and Aspergillus aculeatus, but has a totally different extrolite profile compared to any known Aspergillus species. The type strain of A. saccharolyticus sp. nov. is CBS 127449T ( = IBT 28509T)....
Atta, H.M.; A.S. El-Sayed; M.A. El-Desoukey; Hassan, M.; M. El-Gazar
Natamycin “polyene” antibiotic was isolated from the fermentation broth of a Streptomyces strain No. AZ-55. According to the morphological, cultural, physiological and biochemical characteristics, and 16S rDNA sequence analysis, strain AZ-55 was identified as Streptomyces lydicus. It is active in vitro against some microbial pathogens viz: Staphylococcus aureus, NCTC 7447; Bacillus subtilis, NCTC 1040; Bacillus pumilus, NCTC 8214 ; Micrococcus luteus, ATCC 9341; Escherichia coli, NCTC 10416; ...
Biofouling causes huge economic loss and a recent global ban on organotin compounds as antifouling agents has increased the need for safe and effective antifouling compounds. Five structurally similar compounds were isolated from the crude extract of a marine Streptomyces strain obtained from deep-sea sediments. Antifouling activities of these five compounds and four other structurally-related compounds isolated from a North Sea Streptomyces strain against major fouling organisms were compared to probe structure-activity relationships of compounds. The functional moiety responsible for antifouling activity lies in the 2-furanone ring and that the lipophilicity of compounds substantially affects their antifouling activities. Based on these findings, a compound with a straight alkyl side-chain was synthesized and proved itself as a very effective non-toxic, anti-larval settlement agent against three major fouling organisms. The strong antifouling activity, relatively low toxicity, and simple structures of these compounds make them promising candidates for new antifouling additives. © 2009 Elsevier Ltd. All rights reserved.
Liu, Gang; Chater, Keith F; Chandra, Govind; Niu, Guoqing; Tan, Huarong
Streptomycetes are the most abundant source of antibiotics. Typically, each species produces several antibiotics, with the profile being species specific. Streptomyces coelicolor, the model species, produces at least five different antibiotics. We review the regulation of antibiotic biosynthesis in S. coelicolor and other, nonmodel streptomycetes in the light of recent studies. The biosynthesis of each antibiotic is specified by a large gene cluster, usually including regulatory genes (cluster-situated regulators [CSRs]). These are the main point of connection with a plethora of generally conserved regulatory systems that monitor the organism's physiology, developmental state, population density, and environment to determine the onset and level of production of each antibiotic. Some CSRs may also be sensitive to the levels of different kinds of ligands, including products of the pathway itself, products of other antibiotic pathways in the same organism, and specialized regulatory small molecules such as gamma-butyrolactones. These interactions can result in self-reinforcing feed-forward circuitry and complex cross talk between pathways. The physiological signals and regulatory mechanisms may be of practical importance for the activation of the many cryptic secondary metabolic gene cluster pathways revealed by recent sequencing of numerous Streptomyces genomes.
Dolka, Beata; Olsen, Rikke Heidemann; Thøfner, Ida;
Here, we report five draft genome sequences of Enterococcus cecorum strains that were isolated from different bird species of affected poultry flocks (commercial broilers [CB], broiler breeders [BB], commercial layers [CL], ducks [D], and geese [G]) in Poland....
M. Tidjani Alou; S. Khelaifia; B. La Scola; Cassir, N.
We report the main characteristics of “Lachnoclostridium touaregense” strain Marseille-P2415T (= CSUR P2415 = DSM 102219), a new bacterial species isolated from the gut microbiota of a healthy young girl from Niger.
M. Tidjani Alou; J.-C. Lagier; B. La Scola; Cassir, N.
We report the main characteristics of ‘Lachnoclostridium massiliosenegalense’ strain mt23T (=CSUR P299 =DSM 102084), a new bacterial species isolated from the gut microbiota of a healthy young girl from Senegal.
Samira Kiani; Nafiseh Sadat Naghavi; Alireza Nazari
.... Materials and methods: In the present study Vibrio species isolated from ornamental guppy fish in Kashan, Isfahan, Iran fish ponds and were detected according to molecular detection and genetic alignment...
Zalacain, M; Cundliffe, E
Ribosomes from Streptomyces griseofuscus expressing tlrA, a resistance gene isolated from the tylosin producer Streptomyces fradiae, are resistant to macrolide and lincosamide antibiotics in vitro. The tlrA product was found to be a methylase that introduces two methyl groups into a single base within 23S rRNA, generating N6,N6-dimethyladenine at position 2058. This activity is therefore similar to the ermE resistance mechanism in Saccharopolyspora erythraea (formerly Streptomyces erythraeus). Images PMID:2753855
Olumide Adedokun Odeyemi; Asmat Ahmad
Objective: To investigate antibiotics and heavy metals resistance profile of Aeromonas species isolated from Kuala Gula and Kuala Sepetang, Perak. Methods: Isolated Aeromonas species were subjected to 12 antibiotics: penicillin G 10 units, tetracycline 30 µg, novobiocin 5 µg, nitrofurantoin 100 µg, ampicillin 10 µg, chloramphenicol 10 µg, fluconazole 25 µg, gentamicin 10 µg, streptomycin 10 µg and 25 µg, kanamycin 30 µg, and vancomycin 30 µg and 5 heavy metals: MnSO4.H2O, Cr(NO...
Ariane Bruder-Nascimento; Carlos Henrique Camargo; Alessandro Lia Mondelli; Maria Fátima Sugizaki; Terue Sadatsune; Eduardo Bagagli
Over the last decades, there have been important changes in the epidemiology of Candida infections. In recent years, Candida species have emerged as important causes of invasive infections mainly among immunocompromised patients. This study analyzed Candida spp. isolates and compared the frequency and biofilm production of different species among the different sources of isolation: blood, urine, vulvovaginal secretions and peritoneal dialysis fluid. Biofilm production was quantified in 327 Ca...
Tashiro, Masato; Murakami, Hinako; Yoshizawa, Sadako; Tateda, Kazuhiro; Yamaguchi, Keizo
We evaluated species distribution and antifungal susceptibility of Candida isolates during 2002-2008. Of 177 Candida isolates from blood, species distribution was 90 (51%) Candida albicans, 30 (17%) C. parapsilosis, 22 (12%) C. glabrata, 6 (3%) C. tropicalis and 29 (16%) other Candida spp.. Of 162 Candida isolates from vascular catheter, species distribution was 87 (54%) C. albicans, 14 (9%) C. parapsilosis, 36 (22%) C. glabrata, 5 (3%), C. tropicalis, 2 (1%) C. krusei and 18 (11%) other Candida spp.. Of 1889 Candida isolates from urine, species distribution was 1165 (62%) C. albicans, 22 (1%) C. parapsilosis, 484 (26%) C. glabrata, 83 (4%) C. tropicalis, 26 (1%) C. krusei and 109 (6%) other Candida spp.. Of 782 Candida isolates from stool, species distribution was 425 (54%) C. albicans, 3 (1%) C. parapsilosis, 103 (13%) C. glabrata, 28 (4%) C. tropicalis, 5 (1%), C. krusei and 218 (28%) other Candida spp. Both C. albicans and non-Candida spp. isolated from urine increased slightly over the past 7 years. Flucytosine, fluconazole, itraconazole and micafungin still have strong activity against Candida isolates.
de Vries, J. J. C.; Arents, N. L. A.; Manson, W. L.
Campylobacter species are frequently isolated from fecal specimens of patients with diarrheal illness. Several Campylobacter species are commonly isolated from the oral cavity. In contrast, Campylobacter species are rarely isolated from extra-oro-intestinal abscesses. Reported here are four cases of
Talaromyces species isolated from maize in the U.S., primarily between 1970 and 2014 were grown up from lyophilized storage to identify potential seed endophytes. These isolates had been predominantly identified as Penicillium funiculosum following the taxonomic system of Raper & Thom (1949), althou...
Prakasham Reddy Shetty
Full Text Available A highly potent secondary metabolite producing actinomycetes strain is isolated from marine soil sediments of Visakhapatnam sea coast, Bay of Bengal. Over all ten strains are isolated from the collected soil sediments. Among the ten actinomycetes strains the broad spectrum strain RSPSN2 was selected for molecular characterization, antibiotic production and its purification. The nucleotide sequence of the 1 rRNA gene (1261 base pairs of the most potent strain evidenced a 96% similarity with Streptomyces parvulus 1044 strain, Streptomyces parvulus NBRC 13193 and Streptomyces parvulus BY-F. From the taxonomic features, the actinomycetes isolate RSPSN2 matches with Streptomyces parvulus in the morphological, physiological and biochemical characters. Thus, it was given the suggested name Streptomyces parvulus RSPSN2. The active metabolite was extracted using ethyl acetate (1:3, v/v at pH 7.0. The separation of active ingredient and its purification was performed by using both thin layer chromatography (TLC and column chromatography (CC techniques. Spectrometric studies such as UV-visible, FTIR, and NMR and mass were performed. The antibacterial activity of pure compound was performed by cup plate method against some pathogenic bacteria including of streptomycin resistant bacteria like (Pseudomonas mirabilis. Pseudomonas putida and Bacillus cereus. In conclusion, the collected data emphasized the fact that a polypeptide antibiotic (Actinomycin D was produced by Streptomyces parvulus RSPSN2.
Johnson, Melissa A; Price, Donald K; Price, Jonathan P; Stacy, Elizabeth A
Recent reviews of reproductive isolation (RI) in plants propose that boundaries between closely related species are maintained predominantly through prezygotic mechanisms. However, few experimental studies have explored how boundaries are maintained in long-lived species. Hawaiian Cyrtandra presents an intriguing challenge to our understanding of RI, as it comprises 60 shrub or small tree species that are almost exclusively restricted to wet forests, where sympatry of multiple species is common. We assessed the relative strengths of pre- and postzygotic barriers among four species of Cyrtandra occurring at the extremes of the main Hawaiian Island's natural island-age gradient, Kaua'i (4.7 Myr) and Hawai'i Island (0.6 Myr), to contrast the strengths and stages of reproductive isolation among species at different stages of divergence. A combination of F1 seed germination, F1 seedling survival, and F1 seedling growth isolated (61-91%) three of the species from sympatric relatives. In contrast, the fourth species was isolated (59%) from its sympatric relative through phenological differences alone. Significant postzygotic barriers in between-island crosses were also observed in one species. Results suggest that boundaries between sympatric Cyrtandra species in Hawaii are maintained predominantly through postzygotic barriers. Observations from between-island crosses indicate that postzygotic barriers can arise in allopatry, which may be important in the initial divergence of populations. Future studies of RI in Cyrtandra should include a broader range of species to determine if postzygotic isolating barriers are foremost in the maintenance of species boundaries in this large genus. © 2015 Botanical Society of America.
Full Text Available During the ten year study i.e. 1978 to 1987, out of 53,251 cases of suspected enteric fever, S.typhi was isolated from 4005 cases (7% and S.para typhi ′A′ from 1108 cases (2.5%. A total of 52 strains of S. typhi and 4 strains of S.para typhi ′A′ were found to be resistant to chloramphenicol. Minimal inhibitory concentration was greater than 256 micrograms/ml of all the strains of S.typhi and S.para typhi ′A′.
Braña, Alfredo F; Rodríguez, Miriam; Pahari, Pallab; Rohr, Jurgen; García, Luis A; Blanco, Gloria
Activation and silencing of antibiotic production was achieved in Streptomyces albus J1074 and Streptomyces lividans TK21 after introduction of genes within the thienamycin cluster from S. cattleya. Dramatic phenotypic and metabolic changes, involving activation of multiple silent secondary metabolites and silencing of others normally produced, were found in recombinant strains harbouring the thienamycin cluster in comparison to the parental strains. In S. albus, ultra-performance liquid chromatography purification and NMR structural elucidation revealed the identity of four structurally related activated compounds: the antibiotics paulomycins A, B and the paulomenols A and B. Four volatile compounds whose biosynthesis was switched off were identified by gas chromatography-mass spectrometry analyses and databases comparison as pyrazines; including tetramethylpyrazine, a compound with important clinical applications to our knowledge never reported to be produced by Streptomyces. In addition, this work revealed the potential of S. albus to produce many others secondary metabolites normally obtained from plants, including compounds of medical relevance as dihydro-β-agarofuran and of interest in perfume industry as β-patchoulene, suggesting that it might be an alternative model for their industrial production. In S. lividans, actinorhodins production was strongly activated in the recombinant strains whereas undecylprodigiosins were significantly reduced. Activation of cryptic metabolites in Streptomyces species might represent an alternative approach for pharmaceutical drug discovery.
JIANGChengying; LIUHuizhou; 等
Five bacterial strains,which are able to grow and to disintegrate dibenzothiophene (DBT) and dibenzothiophene sulfone (DBTO2) in fossil fuels,are isolated.Analysis of products of DBT metabolized by these strains shows that different bacteria strains oxidize DBT by different pathways.The isolated strains R-6,R-16,R-9 and R-8 can metabolize DBT to DBTO2 and 2-hydroxybiphenyl(HBP),which are identified as Bacillus brevis,Bacillus sphaericus,Nocardia globerula and Pseudomonas delafieldii respectively.Another strain R-12 identified as Pseudomonas sp. can degrade DBT completely but it cannot produce DBTO2 and HBP. The optimum temperature and initial pH for desulfurization by R-8 are 32℃ and 7.02 respectively and pH of the broth decreases during biodegradation.The growth of strain R-8 with different sulfur-sources indicates that this strain in DBT medium has an induction period of 3 days,which is longer than those with dimethylsulfoxide and MgSO4 media,but the growth rate of the bacterial strain in DBT is higher after the induction.Higher growth and desulfurization rates are observed in the DBT-hexadecane system than in both DBT-ethanol and DBT-dimethylformamide systems.Both strains of R-8 and R-9 also show higher desulfurization activities toward other sulfur-substrates,indicating that they have greater desulfurization potential in application.
Stanley Irobekhian Reuben Okoduwa
Full Text Available The quest for an effective alternative means for effluent treatment is a major concern of the modern-day scientist. Fungi have been attracting a growing interest for the biological treatment of industrial wastewater. In this study, Saccharomycescerevisiae and Torulasporadelbrueckii were isolated from spoiled watermelon and inoculated into different concentrations of effluent. The inoculants were incubated for 21-days to monitor the performance of the isolates by measurement of biochemical oxygen demand (BOD, chemical oxygen demand (COD, nitrates, conductivity, phosphates, sulphates and turbidity. The results showed that Saccharomycescerevisiae had the highest percentage decrease of 98.1%, 83.0%, 60.7%, 60.5%, and 54.2% for turbidity, sulphates, BOD, phosphates and COD, respectively, of the tannery effluent. Torulasporadelbrueckii showed the highest percentage decrease of 92.9%, 90.6%, and 61.9% for sulphates, COD, and phosphates, respectively, while the syndicate showed the highest percentage reduction of 87.4% and 70.2% for nitrate and total dissolve solid (TDS, respectively. The least percentage decrease was displayed by syndicate organisms at 51.2%, 48.1% and 40.3% for BOD, COD and conductivity, respectively. The study revealed that Saccharomycescerevisiae and Torulasporadelbrueckii could be used in the biological treatment of tannery-effluent. Hence, it was concluded that the use of these organisms could contribute to minimizing the adverse environmental risks and health-hazards associated with the disposal of untreated tannery-effluents.
Rainbow trout (Oncorhynchus mykiss) grown in recirculating aquaculture systems (RAS) can acquire “earthy” and “musty” taints due to bioaccumulation of geosmin and 2-methyisoborneol (MIB), respectively, in the fish flesh. Certain species of actinomycetes which produce these compounds are attributed a...
Illnait-Zaragozi, M.T.; Martínez-Machin, G.F.; Fernandez-Andreu, C.M.; Perurena-Lancha, M.R.; Theelen, B.; Boekhout, T.; Meis, J.F.G.M.; Klaassen, C.H.
Cryptococcus isolates from Cuban patients were identified as C. neoformans var. grubii. Although this species has since long been associated with bird droppings, a recent genotyping study provided strong evidence for additional origins of exposure. We sampled different species of trees in Havana,
Illnait-Zaragozi, M.T.; Martínez-Machin, G.F.; Fernandez-Andreu, C.M.; Perurena-Lancha, M.R.; Theelen, B.; Boekhout, T.; Meis, J.F.G.M.; Klaassen, C.H.
Cryptococcus isolates from Cuban patients were identified as C. neoformans var. grubii. Although this species has since long been associated with bird droppings, a recent genotyping study provided strong evidence for additional origins of exposure. We sampled different species of trees in Havana, Cu
Talaromyces sect. Islandici was reexamined to determine the prevalence of isolates from maize and the built environment. Using phenotypic analysis, DNA sequencing and phylogenetic and concordance analysis we discovered and described ten new species in section Islandici and one new species in section...
Full Text Available Aim: Salmonella infections are the leading cause of food-borne infections and can cause gastroenteritis outbreaks worldwide. Salmonella species is defined as inability to lactose fermentation, using citrate as a carbon source, using lysine as nitrate source and forming Hydrogen sulfide (H2S in TSI agar. However, confirmation of false positive results is time consuming and lead to increased costs. The aim of this study is to evaluate the performance of CHROMagar Salmonella (CHROMagar Microbiology, France which is developed for isolation and detection of Salmonella species. Material and Method: For this purpose, among a total of 148 isolates which were isolated from various clinical specimens and stocked at the Central Laboratory of Akdeniz University Hospital, 65 were Salmonella spp., 10 were Pseudomonas aeruginosa, 10 were E. coli, 10 were Acinetobacter baumannii, 10 were Klebsiella pneumoniae, 18 were Morganella morganii, 11 were Citrobacter spp., 5 were Providencia spp., 4 were Aeromonas spp., 5 were Proteus spp. were included in this study. All of the 65 Salmonella spp. isolates apperared with mauve colonies at the CHROMagar Salmonella. Results: E. coli and Klebsiella pnemoniae species were seen as blue, Providencia species were seen as pale-blue; Morganella morganii species were seen as pale-pink, mauve; and Pseudomonas aeruginosa species were seen as pale. Acinebacter baumannii and Aeromonas spp. species were also seen as mauve colonies. Dicussion: CHROMagar Salmonella medium can detect Salmonella species with %100 sensitivity, however there is a need to biochemical or serological confirmation.
Rebecca J Lawton
Full Text Available Freshwater macroalgae from the genus Oedogonium have recently been targeted for biomass applications; however, strains of Oedogonium for domestication have not yet been identified. Therefore, the objective of this study was to compare the performance of isolates of Oedogonium collected from multiple geographic locations under varying environmental conditions. We collected and identified wild-type isolates of Oedogonium from three geographic locations in Eastern Australia, then measured the growth of these isolates under a range of temperature treatments corresponding to ambient conditions in each geographic location. Our sampling identified 11 isolates of Oedogonium that could be successfully maintained under culture conditions. It was not possible to identify most isolates to species level using DNA barcoding techniques or taxonomic keys. However, there were considerable genetic and morphological differences between isolates, strongly supporting each being an identifiable species. Specific growth rates of species were high (>26% day-1 under 7 of the 9 temperature treatments (average tested temperature range: 20.9-27.7°C. However, the variable growth rates of species under lower temperature treatments demonstrated that some were better able to tolerate lower temperatures. There was evidence for local adaptation under lower temperature treatments (winter conditions, but not under higher temperature treatments (summer conditions. The high growth rates we recorded across multiple temperature treatments for the majority of species confirm the suitability of this diverse genus for biomass applications and the domestication of Oedogonium.
Lawton, Rebecca J; de Nys, Rocky; Skinner, Stephen; Paul, Nicholas A
Freshwater macroalgae from the genus Oedogonium have recently been targeted for biomass applications; however, strains of Oedogonium for domestication have not yet been identified. Therefore, the objective of this study was to compare the performance of isolates of Oedogonium collected from multiple geographic locations under varying environmental conditions. We collected and identified wild-type isolates of Oedogonium from three geographic locations in Eastern Australia, then measured the growth of these isolates under a range of temperature treatments corresponding to ambient conditions in each geographic location. Our sampling identified 11 isolates of Oedogonium that could be successfully maintained under culture conditions. It was not possible to identify most isolates to species level using DNA barcoding techniques or taxonomic keys. However, there were considerable genetic and morphological differences between isolates, strongly supporting each being an identifiable species. Specific growth rates of species were high (>26% day-1) under 7 of the 9 temperature treatments (average tested temperature range: 20.9-27.7°C). However, the variable growth rates of species under lower temperature treatments demonstrated that some were better able to tolerate lower temperatures. There was evidence for local adaptation under lower temperature treatments (winter conditions), but not under higher temperature treatments (summer conditions). The high growth rates we recorded across multiple temperature treatments for the majority of species confirm the suitability of this diverse genus for biomass applications and the domestication of Oedogonium.
Singh, Satyendra K; Prasad, Kashi N; Singh, Aloukick K; Gupta, Kamlesh K; Chauhan, Ranjeet S; Singh, Amrita; Singh, Avinash; Rai, Ravi P; Pati, Binod K
Taenia solium is the major cause of taeniasis and cysticercosis/neurocysticercosis (NCC) in the developing countries including India, but the existence of other Taenia species and genetic variation have not been studied in India. So, we studied the existence of different Taenia species, and sequence variation in Taenia isolates from human (proglottids and cysticerci) and swine (cysticerci) in North India. Amplification of cytochrome c oxidase subunit 1 gene (cox1) was done by polymerase chain reaction (PCR) followed by sequencing and phylogenetic analysis. We identified two species of Taenia i.e. T. solium and Taenia asiatica in our isolates. T. solium isolates showed similarity with Asian genotype and nucleotide variations from 0.25 to 1.01 %, whereas T. asiatica displayed nucleotide variations ranged from 0.25 to 0.5 %. These findings displayed the minimal genetic variations in North Indian isolates of T. solium and T. asiatica.
Razafinarivo, Josiane; Jany, Jean-Luc; Crous, Pedro W; Looten, Rachelle; Gaydou, Vincent; Barbier, Georges; Mounier, Jerôme; Vasseur, Valérie
The fungal genus Cladosporium (Cladosporiaceae, Dothideomycetes) is composed of a large number of species, which can roughly be divided into three main species complexes: Cladosporium cladosporioides, Cladosporium herbarum, and Cladosporium sphaerospermum. The aim of this study was to characterize strains isolated from contaminated milk bread rolls by phenotypic and genotypic analyses. Using multilocus data from the internal transcribed spacer ribosomal DNA (rDNA), partial translation elongation factor 1-α, actin, and beta-tubulin gene sequences along with Fourier-transform infrared (FTIR) spectroscopy and morphological observations, three isolates were identified as a new species in the C. sphaerospermum species complex. This novel species, described here as Cladosporium lebrasiae, is phylogenetically and morphologically distinct from other species in this complex.
I. O. Nwankwo
Full Text Available Aim: The study was conducted to determine the prevalence and viability of Campylobacter species isolates from different species of poultry and humans in Sokoto State, Nigeria. Materials and Methods: A cross-sectional study was performed in the live birds markets, humans on admission and at outpatient clinics in the randomly selected hospitals in Sokoto State. Isolation and characterization of Campylobacter species were performed using standard culture isolation techniques and biochemical characterization. A total of 798 (506 cloacal and 292 fecal swabs from poultry and humans, respectively, were collected and analyzed. The viability of 307 isolates stored in 15% glycerol and 85% tryptone broth at −20°C was determined after 7-13 months. Results: A total of 312 (39% were positive for Campylobacter species which comprises 119 (30%, 20 (30%, 3 (14%, 9 (56%, 1 (50%, and 160 (55% in chicken, guinea fowls, pigeons, ducks, turkey, and humans, respectively. The total of 38 (24%, 63 (39%, and 59 (37% humans and 29 (19%, 79 (52%, and 44 (29% poultry isolates were positive for Campylobacter jejuni, Campylobacter Coli, and Campylobacter Lari, respectively. A total of 261 (85% of the stored isolates were still viable on re-isolation with the viability rates of 41 (95%, 67 (85%, and 17 (59% at 7, 9, and 13 months of storage, respectively. There was a negative correlation between months of storage and viability rates. However, there was no significant statistical association (p>0.05 between prevalence rate and species of poultry. Conclusion: Campylobacter species have been detected with varying degree of prevalence in both poultry and humans and their ability to survive freezing at −20°C (95% for up to 7 months has been revealed in the study. This is not only a concern to food and livestock industries but also a concern to the public health at large, especially, in view of the study area being considered one of the largest livestock producers in Nigeria
Full Text Available In this study, attempts were made to isolate Streptomyces sp. from soil samples of two different regions of Bangladesh and evaluate their antagonistic activity against fish and human pathogenic bacteria. A total of 10 isolates were identified as Streptomyces sp. based on several morphological, physiological and biochemical tests. Cross streak method was used to observe the antagonistic activity of the Streptomyces sp. isolates against different fish pathogens belonging to the genus Aeromonas, Pseudomonas and Edwardsiella and human clinical isolates belonging to the genus Klebsiella, Salmonella and Streptococcus. Seven Streptomyces sp. isolates showed antagonism against both fish and human pathogenic bacteria. Four isolates viz., N24, N26, N28 and N47 showed broad spectrum of antagonistic activity (80-100% against all genera of fish and human pathogenic bacteria. The isolate N49 exhibited highest spectrum of antagonism against all fish pathogens (90-100% but comparatively lower degree of antagonism against human pathogens (50-60%. Rest of the two isolates (N21 and N23 showed variability in their antagonism. Results showed that broad spectrum antibiotic(s could be developed from the isolates N24, N26, N28 and N47against several human and fish pathogens. The isolate N49 could be a potential source of antibiotic, especially for fish pathogenic bacteria.
Chung, Yeon Soo; Kwon, Ka Hee; Shin, Sook; Kim, Jae Hong; Park, Yong Ho; Yoon, Jang Won
Possible cross-transmission of hospital-associated enterococci between human patients, medical staff, and hospital environments has been extensively studied. However, limited information is available for veterinary hospital-associated Enterococcus isolates. This study investigated the possibility of cross-transmission of antibiotic-resistant enterococci between dog patients, their owners, veterinary staff, and hospital environments. Swab samples (n =46 5) were obtained from five veterinary hospitals in Seoul, Korea, during 2011. Forty-three Enterococcus strains were isolated, representing seven enterococcal species. E. faecalis and E. faecium were the most dominant species (16 isolates each, 37.2%). Although slight differences in the antibiotic resistance profiles were observed between the phenotypic and the genotypic data, our antibiogram analysis demonstrated high prevalence of the multiple drug-resistant (MDR) isolates of E. faecalis (10/16 isolates, 62.5%) and E. faecium (12/16 isolates, 75.0%). Pulsed-field gel electrophoretic comparison of the MDR isolates revealed three different clonal sets of E. faecalis and a single set of E. faecium, which were isolated from different sample groups or dog patients at the same or two separate veterinary hospitals. These results imply a strong possibility of cross-transmission of the antibiotic-resistant enterococcal species between animal patients, owners, veterinary staff, and hospital environments.
Full Text Available One hundred thirty-six isolates, 88 human and 48 environmental, that met the requirements to belong to the genus Paenibacillus were identified using a polyphasic taxonomic approach known as 16S rRNA plus phenotypic traits. Thirty-seven Paenibacillus species were identified; some had not been previously reported from clinical samples. The main species were P. pabuli (13 isolates, P. provencensis (11, P. phoenicis (9 and P. lautus (8. P. pabuli (11/13 and P. provencensis (8/11 were mainly environmental isolates, while P. phoenicis (9/9 and P. lautus (6/8 were mainly human isolates. Despite the difficulties in assigning to human Paenibacillus isolates a role as a pathogen or contaminant, here 25% of the isolates were involved in true infections, especially in those cases that affected abscesses, wound exudates, ocular infections and diverse fluids. In addition, 15 isolates were identified as 11 ‘Candidatus’ to a new species, all of them from human specimens except one that was obtained from laboratory air. The antimicrobial susceptibility testing showed 95.6% of isolates were resistant to ampicillin, 44% were resistant to cotrimoxazole, 20 to 30% were resistant to cefotaxime and vancomycin and 13% were resistant to rifampicin and erythromycin.
R. B. Faden
Full Text Available The reproductive biology and ecology of five non-hybridizing, sympatric species of Aneilema in Kenya is investigated. The high number of species at the study site is believed to be due to the ecological diversity at this locality. Differences in ecology and seasonal flowering periods appear to be inadequate to reproductively isolate these species. However, differences in daily flowering times, pollinators, floral morphology and opening behaviour, and in chromosome number, alone or in combination (perhaps also coacting with other factors not fully evaluated, are effective isolating mechanisms.
Safar, Silvana Vilas Boas; Gomes, Fátima C O; Marques, Andréa R; Lachance, Marc-André; Rosa, Carlos A
Two isolates of a novel yeast species were obtained from water tanks (phytotelmata) of the bromeliad Vriesea minarum collected in a tableland ('campo rupestre') ecosystem in Brazil. The sequences of the D1/D2 domains of the large-subunit rRNA gene showed that this species is related to Kazachstania exigua and others, from which it differs by 8-10 nucleotide substitutions. The novel species Kazachstania rupicola sp. nov. is proposed to accommodate these isolates. The type strain is UFMG-BRO-80(T) ( = CBS 12684(T) = CBMAI 1466(T)).
Manteca, Angel; Ye, Juanying; Sánchez, Jesús
Streptomycetes are bacterial species that undergo a complex developmental cycle that includes programmed cell death (PCD) events and sporulation. They are widely used in biotechnology because they produce most clinically relevant secondary metabolites. Although Streptomyces coelicolor is one...... events were detected during the presporulation and sporulation stages (80%). Most of these phosphorylations were not reported before in Streptomyces, and included sporulation factors, transcriptional regulators, protein kinases and other regulatory proteins. Several of the identified phosphorylated...
Rajib Kumar Shil
Full Text Available The cellulolytic bacteria from the gut of three different phytophagous insects were studied to isolate novel cellulolytic organism for biofuel industry. Among the threse, gut of P. quatuordecimpunctata larvae contained both highest no of total bacterial count (6.8x107CFU/gut and cellulolytic bacteria (5.42x103CFU/gut. Fifteen different isolates were obtained from the gut of O. velox, A. miliarisand P. quatuordecimpunctata. All the isolates produced clear zone in CMC medium staining with Congo red. The isolates included Gram positive Enterococcus, Microbacterium and Gram negative Aeromonas, Erwinia, Serretia, Flavobacterium, Acenitobacter, Klebsiella, Yersinia, Xenorhabdus, Psedomonas and Photorhabdus. Out of the fifteen isolated and identified bacterial species, twelve bacterial species were novel being reported for first time as having cellulase activity.
Jamuna, M; Jeevaratnam, K
Lactic acid bacteria have received increased attention as a potential food preservative due to their strong antagonistic activity against many food-spoilage and pathogenic organisms. Three Pediococcus species, P. acidilactici NCIM 2292 , P. pentosaceous. NCIM 2296 and P. cervisiae NCIM 2171, were evaluated for bacteriocin production. Inhibitory substance were produced during the late growth phase and maximum production occurred at 37 degrees after 36-48 h of incubation. Bacteriocins partially purified from these species by cold-acetone precipitation at 0 degrees C and cell adsorption desorption techniques have a broad inhibitory spectrum against microorganisms, including gram-negative bacteria such as Escherichia coli and Pseudomonas. Proteolytic enzymes inactivated these peptides, but amylase and lipase did not show any effect. The bacteriocins were stable over a wide pH range (3-8) and apparently most active at pH 4.0-5.0. They were heat-stable (1 h at approximately 80 degrees C and autoclaving) at pH 5.0. No loss in activity was observed when stored under refrigeration (4-8 degrees C). Tris-Tricine SDS-PAGE revealed the molecular masses of these peptides to be between 3.5 and 5.0 kDa.
Stepanova, T F; Bukharin, O V; Kataeva, L V; Perunova, N B; Karpukhina, N F
The purpose of this investigation was to study the species composition and biological properties of Aeromonas bacteria isolated from Bithyniidae mollusks and their habitat (a water reservoir). The Bithyniidae mollusks and water from their habitat were the material to be studied. A total of 176 Aeromonas strains were isolated from the mollusks and water. A. veronii, A. hydrophila, and A. ichthiosmia were most common in the mollusks and A. veronii and A. ichthiosmia were in the water. All the strains isolated had hemolytic activity and no lysozyme or plasma coagulase activity. The magnitude of lecithinase and antilysozymic activities and biofilm formation of the Aeromonas bacteria varied with the isolation source of their strains.
Mustafa Altay Atalay
[¤]CONCLUSION[|] In our hospital, C. albicans was the most frequently isolated Candida species from urine cultures, however, C. glabrata was found as the second most frequent species. As a result, in parallel to the increase of patient population who are at risk for Candida infections, the necessity of doing epidemiological studies for identification of species and susceptibility tests including new antifungal agents was concluded.[¤
Sang, Hyunkyu; An, Tae-Jin; Kim, Chang Sun; Choi, Young Phil; Deng, Jian-Xin; Paul, Narayan Chandra; Sung, Gi-Ho; Yu, Seung Hun
Two isolates of monoverticillate Penicillium species were collected from a grape and schisandra fruit in Korea. Multigene phylogenetic analyses with the nuclear ribosomal internal transcribed spacer (ITS) region and genes encoding β-tubulin (benA) and calmodulin (cmd), as well as morphological analyses revealed that the two isolates are members of the P. sclerotiorum complex in Penicillium subgenus Aspergilloides, but different from species of the P. sclerotiorum complex. The isolates are closely related to P. cainii, P. jacksonii, and P. viticola in terms of their multigene phylogeny, but their colony and conidiophore morphologies differ from those of closely related species. The name P. daejeonium is proposed for this unclassified new species belonging to the P. sclerotiorum complex in subgenus Aspergilloides.
Alenzi, Faris Q.B.
Objective: Fungal urinary tract infections due to Candida have increased significantly in recent years. Our research objective was to study Candida species in urine samples of patients with urinary tract infections (UTIs) associated with obstructive uropathy and to investigate the virulence factors of the isolated Candida. Methods: Patients were divided into two groups: Group I (cases): 50 patients with UTIs and obstructive uropathy. Group II (control): 50 patients with UTIs but with no functional or anatomical obstruction of their urinary tract. Clinical histories and physical examinations, together with laboratory investigations of urine samples were carried out in all patients in this study. Mid stream urine samples were examined microscopically and by fungal cell culture. The isolated Candida species were identified by analytical profile index (API). Candida Virulence factors were determined for the isolated Candida. The susceptibility to fluconazole was evaluated. Results: This study revealed an overall isolation rate of 27% of Candida species among all patient groups. The rate was 36% in cases, and 18% in controls, a difference found to be statistically significant (P<0.05). By API, C.albicans was detected in 44% of Candida species in cases, and in 33% in controls. While C.glabrata was detected in 28% of Candida species in cases, and in 22% in controls. C.tropicalis was detected in 17% of Candida species in cases, and in 22% in controls. Both C.krusei and C.kyfr were detected in 5.5% of Candida species in cases, and in 11% in controls. In terms of virulence factors the study showed that 11 out of 27 (40.5%) of Candida isolates were biofilm positive by tube adherence. Phospholipase activity was demonstrated in 12 out of 27 (44.5%) of Candida isolates. Secretory aspartic proteinase activity was demonstrated in 13 out of 27 (48%) of the Candida isolates. Conclusion: Candida is an important cause of UTIs and obstructive uropathy is a major predisposing factor
Rasimus-Sahari, Stiina; Mikkola, Raimo; Andersson, Maria A; Jestoi, Marika; Salkinoja-Salonen, Mirja
Reasons for mammalian cell toxicity observed in barley and spring wheat grains were sought. Streptomyces sp. isolates from wheat and barley produced heat-stable methanol-soluble substances which inhibited the motility of exposed porcine spermatozoa used as a toxicity indicator. Several barley isolates produced antimycin A (2 to 5 ng/mg wet wt of biomass), a macrolide antibiotic known to block oxygen utilization in mitochondria. The antimycin-producing isolates were members of the Streptomyces albidoflavus group. In in vitro assays with porcine kidney tubular epithelial cells, the specific toxicity of antimycin A towards mitochondria was higher than that of the mycotoxin enniatin B but lower than that of the mitochondriotoxins cereulide and paenilide, produced by food-related Bacillus cereus and Paenibacillus tundrae, respectively. The toxic wheat isolates, related to Streptomyces sedi, did not produce antimycin A and or any other known toxin. Our results suggest that the presence of toxin-producing streptomycetes in stored cereal grains may pose a thus far unrecognized threat for food and feed safety.
Wang, Ching-Min; Lai, Chih-Cheng; Tan, Che-Kim; Huang, Yu-Chuang; Chung, Kuei-Pin; Lee, Meng-Rui; Hwang, Kao-Pin; Hsueh, Po-Ren
Human infections due to Roseomonas species are uncommon and the vast majority of reported infections are opportunistic and easy to treat. We retrospectively reviewed the computerized database of the Bacteriology Laboratory at the National Taiwan University Hospital to identify patients with infections caused by Roseomonas species during the period January 2000 to December 2010. Isolates of Roseomonas species were confirmed by 16S rRNA gene sequencing analysis. During the study period, 20 patients had cultures positive for Roseomonas species. R. mucosa was the most prevalent isolate (n = 18), followed by 1 each of R. gilardii and Roseomonas genomospecies 5. True infection caused by Roseomonas species was confirmed in 17 (85%) patients. Most (n = 12, 71%) of these infections were health care-associated infection. The majority of the patients (n = 12, 71%) had underlying diseases. Malignancy was the most common underlying disease, and catheter-related bloodstream infection was the most common type of infection. The antimicrobial susceptibility patterns varied among the different Roseomonas species. In conclusion, Roseomonas species can cause infection in children and adults regardless of immune status. Because different Roseomonas species may have different clinical features and susceptibility profiles, molecular studies are necessary to identify Roseomonas isolates to the species level.
Gehring, I; Geider, K
Many pathogenic and epiphytic bacteria isolated from apples and pears belong to the genus Erwinia; these include the species E. amylovora, E. pyrifoliae, E. billingiae, E. persicina, E. rhapontici and E. tasmaniensis. Identification and classification of freshly isolated bacterial species often requires tedious taxonomic procedures. To facilitate routine identification of Erwinia species, we have developed a PCR method based on species-specific oligonucleotides (SSOs) from the sequences of the housekeeping genes recA and gpd. Using species-specific primers that we report here, differentiation was done with conventional PCR (cPCR) and quantitative PCR (qPCR) applying two consecutive primer annealing temperatures. The specificity of the primers depends on terminal Single Nucleotide Polymorphisms (SNPs) that are characteristic for the target species. These PCR assays enabled us to distinguish eight Erwinia species, as well as to identify new Erwinia isolates from plant surfaces. When performed with mixed bacterial cultures, they only detected a single target species. This method is a novel approach to classify strains within the genus Erwinia by PCR and it can be used to confirm other diagnostic data, especially when specific PCR detection methods are not already available. The method may be applied to classify species within other bacterial genera.
Tulumoglu, Sener; Yuksekdag, Zehra Nur; Beyatli, Yavuz; Simsek, Omer; Cinar, Berat; Yaşar, Esra
In the present research, the 20 lactobacilli isolated from children feces aged 4-15 years old were investigated for their capabilities to survive at pH 2.0, 2.5, 3.0 and in the presence of 0.25, 0.50 and 0.75% bile salts, their effect on the growth of pathogens, in addition to their sensitivity against 13 selected antibiotics. All the lactobacilli strains were able to survive in low pH and bile salt conditions at pH 2.0 and 0.25% bile salt for 2 h. Moreover, all lactobacilli strains exhibited inhibitory activity against Escherichia coli ATCC 11229, Pseudomonas aeruginosa ATCC 27853 and Staphylococcus aureus ATCC 29213. In addition, all lactobacilli strains indicated resistance to teicoplanin, vancomycin, and bacitracin. The amount of exopolysaccharide (EPS) produced by the strains was 70 and 290 mg/L. The capabilities to autoaggregation and coaggregate with E. coli ATCC 11229 of the strains were also evaluated. High EPS-producing strains indicated significant autoaggregation and coaggregation capability with test bacteria (p lactobacilli could be utilized for preliminary screening in order to identify potentially probiotic bacteria suitable for human.
Zarchi, Ali Akbar Karimi; Vatani, Hadi
The presence of cockroaches has health implications, such as nosocomial infection, as the insects move freely from areas within and around hospitals that may harbor pathogenic organisms. The goals of the present study were to determine species of bacteria isolated from cockroaches found in three Tehran hospitals, and to determine distribution of potential vectors by species and sex. The study is descriptive laboratory research. A total of 305 cockroaches from three species were trapped and identified: Periplaneta americana (65.6%), Blatella germanica (12.1%), and Blatta orientalis (22.3%). From these potential vectors, 19 species of bacteria were isolated and identified. After transportation of cockroaches to the laboratory, separation of the whole-homogenized suspension of each species was carried out. Identification of the isolated bacteria was performed according to Burgey's Manual of Determinative Bacteriology, 9th Edition. The most common species of bacteria isolated from cockroaches were Escherichia coli, Streptococcus Group D, Bacillus spp., Klebsiella pneumoniae, and Proteus vulgaris. No statistical significance was found between sex and species of cockroach carrying bacteria (p > 0.05), but significance was found for sex in Citrobacter freundii, Staphylococcus aureus, and Streptococcus non-group A&B (p < 0.05).
Isoken H. Igbinosa
Full Text Available This study assessed the prevalence of antibiotic-resistant Aeromonas species isolated from Alice and Fort Beaufort wastewater treatment plant in the Eastern Cape Province of South Africa. Antibiotic susceptibility was determined using the disc diffusion method, and polymerase chain reaction (PCR assay was employed for the detection of antibiotics resistance genes. Variable susceptibilities were observed against ciprofloxacin, chloramphenicol, nalidixic acid, gentamicin, minocycline, among others. Aeromonas isolates from both locations were 100% resistant to penicillin, oxacillin, ampicillin, and vancomycin. Higher phenotypic resistance was observed in isolates from Fort Beaufort compared to isolates from Alice. Class A pse1 β-lactamase was detected in 20.8% of the isolates with a lower detection rate of 8.3% for blaTEM gene. Class 1 integron was present in 20.8% of Aeromonas isolates while class 2 integron and TetC gene were not detected in any isolate. The antibiotic resistance phenotypes observed in the isolates and the presence of β-lactamases genes detected in some isolates are of clinical and public health concern as this has consequences for antimicrobial chemotherapy of infections associated with Aeromonas species. This study further supports wastewater as potential reservoirs of antibiotic resistance determinants in the environment.
Full Text Available Over the last decades, there have been important changes in the epidemiology of Candida infections. In recent years, Candida species have emerged as important causes of invasive infections mainly among immunocompromised patients. This study analyzed Candida spp. isolates and compared the frequency and biofilm production of different species among the different sources of isolation: blood, urine, vulvovaginal secretions and peritoneal dialysis fluid. Biofilm production was quantified in 327 Candida isolates obtained from patients attended at a Brazilian tertiary public hospital (Botucatu, Sao Paulo. C. albicans ALS3 gene polymorphism was also evaluated by determining the number of repeated motifs in the central domain. Of the 198 total biofilm-positive isolates, 72 and 126 were considered as low and high biofilm producers, respectively. Biofilm production by C. albicans was significantly lower than that by non-albicans isolates and was most frequently observed in C. tropicalis. Biofilm production was more frequent among bloodstream isolates than other clinical sources,in urine, the isolates displayed a peculiar distribution by presenting two distinct peaks, one containing biofilm-negative isolates and the other containing isolates with intense biofilm production. The numbers of tandem-repeat copies per allele were not associated with biofilm production, suggesting the evolvement of other genetic determinants.
Bruder-Nascimento, Ariane; Camargo, Carlos Henrique; Mondelli, Alessandro Lia; Sugizaki, Maria Fátima; Sadatsune, Terue; Bagagli, Eduardo
Over the last decades, there have been important changes in the epidemiology of Candida infections. In recent years, Candida species have emerged as important causes of invasive infections mainly among immunocompromised patients. This study analyzed Candida spp. isolates and compared the frequency and biofilm production of different species among the different sources of isolation: blood, urine, vulvovaginal secretions and peritoneal dialysis fluid. Biofilm production was quantified in 327 Candida isolates obtained from patients attended at a Brazilian tertiary public hospital (Botucatu, Sao Paulo). C. albicans ALS3 gene polymorphism was also evaluated by determining the number of repeated motifs in the central domain. Of the 198 total biofilm-positive isolates, 72 and 126 were considered as low and high biofilm producers, respectively. Biofilm production by C. albicans was significantly lower than that by non-albicans isolates and was most frequently observed in C. tropicalis. Biofilm production was more frequent among bloodstream isolates than other clinical sources, in urine, the isolates displayed a peculiar distribution by presenting two distinct peaks, one containing biofilm-negative isolates and the other containing isolates with intense biofilm production. The numbers of tandem-repeat copies per allele were not associated with biofilm production, suggesting the evolvement of other genetic determinants.
廖昌珑; 李文鹏; 刘士清; 王启方
从三七根际分离到一株产木聚糖酶链霉菌Streptomyces sp.YNUCC0233.该菌所产生的木聚糖酶的最适反应温度为67℃,最适pH为6.0,在pH 5.0 ～ 9.0和60℃以下时相对稳定.Ba2+和K+对该木聚糖酶有较强的促进作用,Hg2+、Cu2+、 Mn2+、Co2+、Ni2+和EDTA对该酶有较强的抑制作用.对Streptomyces sp.YNUCC0233 16S rDNA的1105bp片段的序列分析结果表明,Streptomyces sp.YNUCC0233的16S rDNA片段与GenBank中所有已注册的链霉菌分类单位均不同.
Kuznetsov, V D; Filippova, S N; Poltorak, V A
Culture 78 was proposed as a neotype of Streptomyces ruber. It was isolated from the soils of the Baikal region and was closest, in its taxonomic properties, to the original description of the species  whose representative had been lost. Cultures from different microbial collections designated as S. ruber were shown to be unlike the original description. The neotype had the following taxononic properties: the cell wall of type I; spiral sporophores with extended spirals having 2-3 coils; oval spores with a smooth envelope; greyish pink aerial and dark-red substrate mycelia; a red pigment not passing into the medium; slow gelatin liquefaction and milk peptonization; weak starch hydrolysis; assimilation of glucose, xylose, rammose, fructose, and inositol; weak growth on arabinose, raffinose and mannitol, but not on sucrose; no formation of melanoid pigments; synthesis of riboflavin and prodigiosin pigments; inhibition of Gram-positive bacterial and acid-resistant mycobacterial growth; no inhibition of yeast and fungal growth. The culture was sensitive to streptomycin, neomycin, gentamycin, monomycin, tetracycline,erythromycin, oleandomycin, lincomycin, ristomycin, levomycetin, polymyxin and fusidin, but resistant in penicillin. The population was composed of six variants : main, faded, asporogenic red, asporogenic yellow, asporogenic white and nocardia-like. The latter two were not capable of riboflavin and prodigiosin formation. The asporogenic yellow variant was a monosynthetic organism: it formed riboflavin, but could not synthesize prodigiosin. The neotype of S. ruber 78 is deposited withthe national Collection of Microorganisms (the reference number is VKM A-611).
Cuesta, Gonzalo; García-de-la-Fuente, Rosana; Abad, Manuel; Fornes, Fernando
The search for new biocontrol strategies to inhibit the growth of phytopathogenic microorganisms has become widely widespread due to environmental concerns. Among actinomycetes, Streptomyces species have been extensively studied since they have been recognized as important sources of antibiotics. Actinomycete strains were isolated from a calcareous soil, 2 two-phase olive mill waste ('alperujo') composts, and the compost-amended soil by using selective media, and they were then co-cultured with 5 phytopathogenic fungi and 1 bacterium to perform an in vitro antagonism assay. Forty-nine actinomycete strains were isolated, 12 of them showing a great antagonistic activity towards the phytopathogenic microorganisms tested. Isolated strains were identified by 16S rDNA sequence analysis and phenotypic procedures. Eleven isolates concerned the genus Streptomyces and 1 actinomycete with chitinolytic activity belonged to the genus Lechevalieria.
Priscilla de Laet Sant'Ana
Full Text Available Oropharyngeal candidiasis continues to be considered the most common opportunistic disease in Aids patients. This study was designed to investigate species distribution, serotype and antifungal susceptibility profile among Candida spp. isolated from the oral cavity of Aids patients recruited from six Brazilian university centers. Oral swabs from 130 Aids patients were plated onto CHROMagar Candida medium and 142 isolates were recovered. Yeast isolates were identified by classical methods and serotyped using the Candida Check® system-Iatron. Antifungal susceptibility testing was performed according to the NCCLS microbroth assay. C. albicans was the most frequently isolated species (91%, and 70% of the isolates belonged to serotype A. We detected 12 episodes of co-infection (9%, including co-infection with both serotypes of C. albicans. Non-albicans species were isolated from 12 episodes, 50% of them exhibited DDS or resistance to azoles. Otherwise, only 8 out 130 isolates of C. albicans exhibited DDS or resistance to azoles. Brazilian Aids patients are infected mainly by C. albicans serotype A, most of them susceptible to all antifungal drugs.
Toledo, A.; Anda, P.; Escudero, R.; Larsson, C.; Bergstrom, S.; Benach, J. L.
Multilocus sequence analysis (MLSA) was used to clarify the taxonomic status of a virulent Borrelia organism previously isolated from patients with relapsing fever and from ticks in Spain that is designated the Spanish relapsing fever (SRF) Borrelia. This species has been used extensively in experimental infection models because of its continued virulence. Seven genes were amplified to analyze the phylogenetic relationships among several Spanish isolates of SRF Borrelia and other relapsing fever Borrelia species. The genes targeted in this study included rrs and flaB, which have commonly been used in phylogenetic studies; the rrf-rrl intergenic spacer (IGS), which is highly discriminatory; and four additional genes, p66, groEL, glpQ, and recC, which are located on the chromosome and which have therefore evolved in a clonal way. The species included in this study were Borrelia duttonii, B. recurrentis, B. crocidurae, and B. hispanica as Old World Borrelia species and B. turicatae and B. hermsii as New World Borrelia species. The results obtained by MLSA of the SRF Borrelia on the basis of 1% of the genomic sequence data analyzed confirmed that the SRF Borrelia isolates are B. hispanica. However, the prototype isolates of B. hispanica used in this study have an uncertain history and display unique phenotypic characteristics that are not shared with the SRF Borrelia. Therefore, we propose to use strain SP1, isolated from a relapsing fever patient in 1994 in southern Spain, as the type strain for B. hispanica. PMID:20463158
Shaykh Baygloo, Nima; Bouzari, Majid; Rahimi, Fateh; Abedini, Fereydoon; Yadegari, Sima; Soroushnia, Mohsen; Beigi, Fahimeh
The worldwide emergence of multi-drug resistant (MDR) bacteria in recent years has caused many problems for hospitals and patients, especially intensive care unit patients. Among these clinically important MDR bacteria are Acinetobacter baumannii complex species (A. baumannii, Acinetobacter genomic species 3 and Acinetobacter genomic species 13TU) that cause a wide range of infections. The sequencing and bioinformatics analysis of a part of the Zone 1 of rpoB gene was performed for species identification of Acinetobacter isolates obtained from ICU patients with infected burns hospitalized in a hospital in Isfahan, Iran, over a 9-month period. Antibiotic sensitivity of Acinetobacter isolates was investigated using the disk diffusion method and different classes of antibiotics including amikacin, cefotaxime, ceftriaxone, ciprofloxacin, imipenem and piperacillin. Acinetobacter spp. were isolated from 10 of 80 (12.5%) investigated patients. All of the 10 Acinetobacter isolates were identified as Acinetobacter baumannii and multi-drug resistant according to antibiotic susceptibility tests. Of the Acinetobacter baumannii complex members, only A. baumannii species was identified among the isolates obtained from patients with infected burns in an Isfahan hospital over a 9-month period.
Mohan das, Vinitha; Ballal, Mamatha
The number of nosocomial blood stream infections due to Candida species has increased over the past few decades. In order to establish an infection, opportunistic pathogens have to evade the immune system, survive, divide in the host environment, and spread to other tissues. Proteinase and phospholipase secretion has been implicated as potential virulence factors for some Candida species responsible for catheter related candidemia in intensive care unit (ICU) patients with indwelling devices. We therefore have aimed at demonstrating the secretion of proteinase and phospholipase enzymes as virulent factors by Candida species isolated from blood samples collected from ICUs, dialysis units and oncology units. One hundred and fourteen isolates of Candida species were obtained from the blood samples and the isolates include 37 Candida albicans, 7 Candida glabrata, 5 Candida guilliermondii, 3 Candida kefyr, 45 Candida krusei, 5 Candida parapsilosis, and 12 Candida tropicalis. Proteinase assay was performed by using the Staib et al method. Phospholipase assay was performed by using the method of Samaranayake et al. Precipitation zone (Pz value) was determined. The percentage of isolates which produced detectable amounts of proteinase is 74.56% and 44.73% of isolates produced detectable amounts of phospholipase. We believe that production of both phospholipase and proteinase enzimes could be an important virulence factor for several Candida species.
Nasr, Shaghayegh; Soudi, Mohammad Reza; Nasrabadi, Seyyedeh Maryam Zamanzadeh; Nikou, Mahdi Moshtaghi; Salmanian, Ali Hatef; Nguyen, Hai D T
A novel species of basidiomycetes was isolated from kitchen garden soil in Shahryar city, Tehran province, Iran. Molecular and conventional methods were employed to identify and classify this single isolate. Morphologically, the isolate was considered yeast-like with hyaline and oval cells reproducing by monopolar budding, forming ballistoconidia, hyphae, arthroconidia and didymospores. Basidia and basidiospores resembling those produced by Basidioascus species were observed. Sequencing and Bayesian phylogenetic analysis of rRNA genes and the internal transcribed spacer region revealed its sister relationship to described species of the genus Basidioascus. Assimilation and fermentation tests, cell-wall carbohydrate analysis and enzyme activity tests were performed to provide insight into the metabolism of the isolate. Based on morphology, physiology and phylogeny of rRNA gene sequences, the isolate was shown to represent a novel species of the genus Basidioascus, described as Basidioascus persicus sp. nov. (holotype IBRC P1010180(T) = ex-type IBRC M30078(T) = isotype CBS 12808(T)). The MycoBank number of the novel species is MB 804703. An emended description of the genus Basidioascus is also provided.
Zhonglai Luo; Tingting Duan; Shuai Yuan; Shi Chen; Xiufeng Bai; Dianxiang Zhang
Reproductive isolation defines the biological species concept and plays a key role in the formation and maintenance of species. The relative contributions of different isolating stages has been suggested to be closely associated with phylogenetic relatedness. Few studies have focused on the relative contributions of pre- versus post-zygotic mechanisms, and even fewer have been conducted under strict phylogenetic frameworks. Pre-and post-zygotic reproductive isolation stages have been investigated in the sister species Mussaenda kwangtungensis and M. pubescens var. alba. The two species have partly overlapping distribu-tion ranges and flowering times, while the principal pollinators differed strikingly for them, demonstrating strong pre-zygotic isolations. Natural hybrids were detected by simple sequence repeat markers and their maternal parents were identified based on chloroplast gene sequen-ces. Five out of 81 individuals were suggested to be hybrids that fall into the categories F2, BC1, and BC2 by the NewHybrids analysis. Interspecific crossings resulted in significantly reduced fruit set and seed germination rates. Phylogenetic analysis revealed short Kimura-2-parameter distance between M. kwangtungensis and M. pubescens var. alba. These findings strongly supported the hypothesis that for species with a closer phylogenetic relationship, pre-zygotic isolation plays an important part in limiting gene exchange in sympatric areas.
邢莹莹; 吕典秋; 魏琪; 万书明; 董学志; 邱彩玲; 金光辉
为明确黑龙江省马铃薯疮痂病病原菌的种类及其特征,2012-2013年从黑龙江省克山县、绥化市、哈尔滨市、杜尔伯特蒙古族自治县采集具有疮痂病病斑的马铃薯块茎,从中分离纯化病原菌,根据16S rRNA基因的差异采用分子手段对所分离的菌株进行种类和致病性鉴定,并对txtAB阳性菌株采用萝卜幼苗法或马铃薯致病性试验测定其致病性.共分离出74株菌株,鉴定出致病性菌株26株,其中Streptomyces scabies或S.europaeiscabiei 21株,S.turgidiscabies 3株和S.acidiscabies 2株.所有的致病性菌株中共有4种致病岛基因型,即nec1 +/tomA+、nec1-/tomA+、nec 1+/tomA-和nec1-/tomA.
McDonald, Bradon R; Currie, Cameron R
Lateral gene transfer (LGT) profoundly shapes the evolution of bacterial lineages. LGT across disparate phylogenetic groups and genome content diversity between related organisms suggest a model of bacterial evolution that views LGT as rampant and promiscuous. It has even driven the argument that species concepts and tree-based phylogenetics cannot be applied to bacteria. Here, we show that acquisition and retention of genes through LGT are surprisingly rare in the ubiquitous and biomedically important bacterial genus Streptomyces Using a molecular clock, we estimate that the Streptomyces bacteria are ~380 million years old, indicating that this bacterial genus is as ancient as land vertebrates. Calibrating LGT rate to this geologic time span, we find that on average only 10 genes per million years were acquired and subsequently maintained. Over that same time span, Streptomyces accumulated thousands of point mutations. By explicitly incorporating evolutionary timescale into our analyses, we provide a dramatically different view on the dynamics of LGT and its impact on bacterial evolution.IMPORTANCE Tree-based phylogenetics and the use of species as units of diversity lie at the foundation of modern biology. In bacteria, these pillars of evolutionary theory have been called into question due to the observation of thousands of lateral gene transfer (LGT) events within and between lineages. Here, we show that acquisition and retention of genes through LGT are exceedingly rare in the bacterial genus Streptomyces, with merely one gene acquired in Streptomyces lineages every 100,000 years. These findings stand in contrast to the current assumption of rampant genetic exchange, which has become the dominant hypothesis used to explain bacterial diversity. Our results support a more nuanced understanding of genetic exchange, with LGT impacting evolution over short timescales but playing a significant role over long timescales. Deeper understanding of LGT provides new
Xing Shan TIAN; Shuang Da XIE; Xue Bing JIANG; Xiao Mao ZHOU; Li Mei YANG; Ding Jun XIAO
A new cyclic octapeptide, bandunamide, was isolated from the acetone extracts of streptomyces griseovariabilis bandungensis. This cyclic octapeptide exhibits strong antimicrobial activity against Phytophthora drechsleri (IC50=15 ng/mL), Colletotrchum higginsiannum (IC50=15.6 ng/mL), Piricularia oxyzae (IC50=0.2 (g/mL), and Fusarium oxysporum f. Sp. (IC50=100 (g/mL). The structure elucidation of bandunamide is herein reported.
Stabili, L; Gravili, C; Tredici, S M; Piraino, S; Talà, A; Boero, F; Alifano, P
Luminous bacteria are isolated from both Hydrozoa and Bryozoa with chitinous structures on their surfaces. All the specimens of the examined hydroid species (Aglaophenia kirchenpaueri, Aglaophenia octodonta, Aglaophenia tubiformis, Halopteris diaphana, Plumularia setacea, Ventromma halecioides), observed under blue light excitation, showed a clear fluorescence on the external side of the perisarc (chitinous exoskeleton) around hydrocladia. In the bryozoan Myriapora truncata, luminous bacteria are present on the chitinous opercula. All the isolated luminous bacteria were identified on the basis of both phenotypic and genotypic analysis. The isolates from A. tubiformis and H. diaphana were unambiguously assigned to the species Vibrio fischeri. In contrast, the isolates from the other hydroids, phenotypically assigned to the species Vibrio harveyi, were then split into two distinct species by phylogenetic analysis of 16S rRNA gene sequences and DNA-DNA hybridization experiments. Scanning electron microscopy analysis and results of culture-based and culture-independent approaches enabled us to establish that luminous vibrios represent major constituents of the bacterial community inhabiting the A. octodonta surface suggesting that the interactions between luminous bacteria and the examined hydrozoan and bryozoan species are highly specific. These interactions might have epidemiological as well as ecological implications because of the opportunistic pathogenicity of luminous Vibrio species for marine organisms and the wide-distribution of the hydrozoan and bryozoan functioning as carriers.
Olumide Adedokun Odeyemi; Asmat Ahmad
Objective: To investigate antibiotics and heavy metals resistance profile of Aeromonas species isolated from Kuala Gula and Kuala Sepetang, Perak. Methods: Isolated Aeromonas species were subjected to 12 antibiotics: penicillin G 10 units, tetracycline 30 µg, novobiocin 5 µg, nitrofurantoin 100 µg, ampicillin 10 µg, chloramphenicol 10 µg, fluconazole 25 µg, gentamicin 10 µg, streptomycin 10 µg and 25 µg, kanamycin 30 µg, and vancomycin 30 µg and 5 heavy metals: MnSO4.H2O, Cr(NO3).9H2O, CuSO4, Pb(NO3)2, HgCl2. Results: Results from this study revealed that isolates from the two sources-displayed varied resistance. Isolates from Kuala Sepetang showed 91%, 66%, 61%, and 52% resistance to vancomycin, Novobiocin, streptomycin and gentamycin while Kuala Gula isolates showed 76%, 93%, 28% and 14% respectively. High rate of resistance was shown to be against lead (Pb), Chromium (Cr), copper (Cu) and Manganese (Mn) with minimum inhibitory concentration ranging from 2 560 µg/mL to>5120 µg/mL. Low resistance to mercury was observed among isolates from Kuala Gula. Conclusions: This study showed that there was widespread of both heavy metals and antibiotics resistance among Aeromonas isolates from the two marine sources with isolates from Kuala Sepetang showing more resistance thereby posing danger to both aquatic animals and human health.
Olumide Adedokun Odeyemi
Full Text Available Objective: To investigate antibiotics and heavy metals resistance profile of Aeromonas species isolated from Kuala Gula and Kuala Sepetang, Perak. Methods: Isolated Aeromonas species were subjected to 12 antibiotics: penicillin G 10 units, tetracycline 30 µg, novobiocin 5 µg, nitrofurantoin 100 µg, ampicillin 10 µg, chloramphenicol 10 µg, fluconazole 25 µg, gentamicin 10 µg, streptomycin 10 µg and 25 µg, kanamycin 30 µg, and vancomycin 30 µg and 5 heavy metals: MnSO4.H2O, Cr(NO3.9H2O, CuSO4, Pb(NO32, HgCl2. Results: Results from this study revealed that isolates from the two sources-displayed varied resistance. Isolates from Kuala Sepetang showed 91%, 66%, 61%, and 52% resistance to vancomycin, Novobiocin, streptomycin and gentamycin while Kuala Gula isolates showed 76%, 93%, 28% and 14% respectively. High rate of resistance was shown to be against lead (Pb, Chromium (Cr, copper (Cu and Manganese (Mn with minimum inhibitory concentration ranging from 2 560 µg/mL to >5 120 µg/mL. Low resistance to mercury was observed among isolates from Kuala Gula. Conclusions: This study showed that there was widespread of both heavy metals and antibiotics resistance among Aeromonas isolates from the two marine sources with isolates from Kuala Sepetang showing more resistance thereby posing danger to both aquatic animals and human health.
Jose C Huguet-Tapia
Full Text Available PAISt is a large genomic island located in the chromosome of the plant pathogen Streptomyces turgidiscabies Car8. The island carries clustered virulence genes, transfers to other Streptomyces species, and integrates by site-specific recombination at the 8 bp palindrome TTCATGAA. The palindrome is located at the 3' end of the bacitracin resistance gene (bacA. We demonstrate that PAISt is able to excise in modules by recombination of one internal and two flanking palindromic direct repeats. The gene intSt located at the 3( end of PAISt encodes a tyrosine recombinase. Site-specific recombination activity of intSt was tested and confirmed by heterologous expression in Streptomyces coelicolor. Comparative analysis of PAISt homologues in Streptomyces scabies 87-22 and Streptomyces acidiscabies 84-104 indicates that these islands have been fixed by sequence erosion of intSt and the recombination sites.
Mikkelsen, Deirdre; Milinovich, Gabriel J; Burrell, Paul C; Huynh, Sharnan C; Pettett, Lyndall M; Blackall, Linda L; Trott, Darren J; Bird, Philip S
Porphyromonas species are frequently isolated from the oral cavity and are associated with periodontal disease in both animals and humans. Black, pigmented Porphyromonas spp. isolated from the gingival margins of selected wild and captive Australian marsupials with varying degrees of periodontal disease (brushtail possums, koalas and macropods) were compared phylogenetically to Porphyromonas strains from non-marsupials (bear, wolf, coyote, cats and dogs) and Porphyromonas gingivalis strains from humans using 16S rRNA gene sequence analysis. The results of the phylogenetic analysis identified three distinct groups of strains. A monophyletic P. gingivalis group (Group 1) contained only strains isolated from humans and a Porphyromonas gulae group (Group 2) was divided into three distinct subclades, each containing both marsupial and non-marsupial strains. Group 3, which contained only marsupial strains, including all six strains isolated from captive koalas, was genetically distinct from P. gulae and may constitute a new Porphyromonas species.
Perrone, Giancarlo; Varga, János; Susca, Antonia
A novel species, Aspergillus uvarum sp. nov., is described within Aspergillus section Nigri. This species can be distinguished from other black aspergilli based on internal transcribed spacers (ITS), beta-tubulin and calmodulin gene sequences, by AFLP analysis and by extrolite profiles. Aspergillus...... uvarum sp. nov. isolates produced secalonic acid, common to other Aspergillus japonicus-related taxa, and geodin, erdin and dihydrogeodin, which are not produced by any other black aspergilli. None of the isolates were found to produce ochratoxin A. The novel species is most closely related to two...... atypical strains of Aspergillus aculeatus, CBS 114.80 and CBS 620.78, and was isolated from grape berries in Portugal, Italy, France, Israel, Greece and Spain. The type strain of Aspergillus uvarum sp. nov. is IMI 388523(T)=CBS 127591(T)= ITEM 4834(T)= IBT26606(T)....
Partha Pratim Das
Results: From the 59 culture positive HIV seropositive cases, 61 Candida isolates were recovered; Candidaalbicans (n=47, 77.0%, C. dubliniensis (n=9, 14.7%, C. parapsilosis (n=2, 3.2%, C. glabrata (n=2, 3.2%, and C. famata (n=1, 1.6%. Candida colonization in HIV-seropositive individuals was significantly higher than that of HIV-seronegative (control group. Antifungal susceptibility testing revealed (n=6, 9.3% C. albicans isolates resistant to voriconazole and fluconazole by disk-diffusion method whereas no resistance was seen by Fungitest method. Interpretation & conclusions: C. albicans was the commonest Candida species infecting or colonizing HIV seropositive individuals. Oropharyngeal Candida isolates had high level susceptibility to all the major antifungals commonly in use. Increased level of immunosuppression in HIV-seropositives and drug resistance of non-albicans Candida species makes identification and susceptibility testing of Candida species necessary in different geographical areas of the country.
Clark, Laura C.; Seipke, Ryan F.; Prieto, Pilar; Willemse, Joost; van Wezel, Gilles P.; Hutchings, Matthew I.; Hoskisson, Paul A.
Understanding the evolution of virulence is key to appreciating the role specific loci play in pathogenicity. Streptomyces species are generally non-pathogenic soil saprophytes, yet within their genome we can find homologues of virulence loci. One example of this is the mammalian cell entry (mce) locus, which has been characterised in Mycobacterium tuberculosis. To investigate the role in Streptomyces we deleted the mce locus and studied its impact on cell survival, morphology and interaction with other soil organisms. Disruption of the mce cluster resulted in virulence towards amoebae (Acanthamoeba polyphaga) and reduced colonization of plant (Arabidopsis) models, indicating these genes may play an important role in Streptomyces survival in the environment. Our data suggest that loss of mce in Streptomyces spp. may have profound effects on survival in a competitive soil environment, and provides insight in to the evolution and selection of these genes as virulence factors in related pathogenic organisms. PMID:23346366
Schulte Fischedick, Frederique B; Stuckey, Matthew J; Aguilar-Setién, Alvaro; Moreno-Sandoval, Hayde; Galvez-Romero, Guillermo; Salas-Rojas, Mónica; Arechiga-Ceballos, Nidia; Overgaauw, Paul A M; Kasten, Rickie W; Chomel, Bruno B
Bartonella species are highly endemic among wild rodents in many parts of the world. Blood and/or blood clot cultures from 38 rodents, including 27 Yucatan deer mouse (Peromyscus yucatanicus), 7 Gaumer's spiny pocket mouse (Heteromys gaumeri), 2 black rats (Rattus rattus) and 2 big-eared climbing rats (Ototylomys phyllotis) captured near Merida, Yucatan, Mexico, led to the isolation in 3-4 days of small gram-negative bacilli, which were identified as Bartonella spp. based on colony morphology. DNA extraction and PCR testing were also performed from heart samples of 35 of these 38 rodents. Overall, Bartonella spp. were isolated from the blood/blood clots of 22 (58%) rodents. All Bartonella-positive rodents were Yucatán deer mice from San José Pituch. Sequencing of a fragment of the gltA gene showed that all but one rodent isolates were closest to B. vinsonii subsp. vinsonii and one isolate was intermediate between B. vinsonii subsp. berkhoffii and B. vinsonii subsp. arupensis. Further analysis of concatenated housekeeping genes (gltA, ftsZ, rpoB, and 16S rRNA) suggests that this outlier isolate is a new subspecies within the B. vinsonii genogroup, for which we proposed the name B. vinsonii subsp. yucatanensis.
Wilson, K E; Kempf, A J; Liesch, J M; Arison, B H
Two new carbapenem antibiotics, northienamycin and 8-epi-thienamycin have been isolated from culture broth of Streptomyces cattleya grown under conditions for thienamycin production. The isolation, structure elucidation and in vitro antibacterial spectra of the new carbapenems are reported. In addition, comparison of the in vitro potency of the corresponding formamidine derivatives to that of MK787 is presented.
Iroha Ifeanyichukwu; Ejikeugwu Chika; Anioketta Ogonna; Iroha Chidinma; Ajah Monique; Moses Ikechukwu; Eluu Stanley; Nwakaeze Emmanuel; Afiukwa Ngozi; Nwuzo Agabus
Objective: This study evaluated the occurrence and antimicrobial susceptibility profile of Salmonella species isolated from various poultry products including chicken meat, poultry eggs, poultry bird's drinking water, and poultry feed. Materials and methods: A total of 79 samples comprising of chicken meat (n=20), egg shell (n=15), poultry egg contents (n=18), drinking water (n=14), and poultry feed (n=12) were bacteriologically and microscopically analyzed for the isolation of Salmonell...
Habu A. Kalshingi
Full Text Available Biochemical and molecular analysis were conducted on 34 strains of Mycoplasma species isolated between 2003 and 2009 from the genital tract of clinically healthy Dorper sheep and sheep with ulcerative vulvitis and balanitis. Earlier publications identified the causative agent as Mycoplasma mycoides mycoides large colony (MmmLC and Arcanobacterium pyogenes. The aims of the study were to characterise Mycoplasma species isolated from the genital tract of Dorper sheep with polymerase chain reaction assay, cloning and gene sequencing. Basic Local Alignment Search Tool (BLAST results revealed six predominant Mycoplasma species: Mycoplasma arginini, Mycoplasma bovigenitalium, Arcanobacterium laidlawii, MmmLC, Mycoplasma sp. ovine/caprine serogroup II and M. canadense. Sequencing of the 34 isolates were analysed using phylogenetic methods, and 18 (50% were identified as M. arginini with 99% – 100% similarity to M. arginini from England and Sweden. Six isolates showed 99% similarity to M. bovigenitalium strains from Turkey and Germany. Two isolates had 99% similarity to an M. sp. ovine/caprine sero group II from the United Kingdom. BLAST for two isolates revealed 99% similarity to Acholeplasma laidlawii from India, another two were 99% similar to MmmLC strain from Sweden, two showed 98% similarity to Mycoplasma sp. Usp 120 from Brazil, and two isolates have a 97% – 99% similarity to M. mm. Jcv1 strain from the United States of America. Finally, one isolate showed similarity of 99% to Mycoplasma canadense strain from Italy. The findings support the hypothesis that ulcerative vulvitis and balanitis of Dorper sheep in South Africa (SA is a multifactorial disease with involvement of different Mycoplasma species.
Full Text Available Background and objectives: Bacteria, most prevalently the Pseudomonas species possess high capacity to utilize and degrade"npetroleum hydrocarbons and are classified as the hydrocarbonoclastic microorganisms. Many species of the genus"nPseudomonas are notorious for their aerobic degradation capacity, extracellular enzyme production and are metabolically"nversatile organisms capable of utilizing a wide range of hydrocarbons and other compounds. In this study, the ability of diesel"nutilization by some locally isolated Pseudomonas species was tested."nMaterials and Methods: From a local red laterite soil, four different Pseudomonas species were isolated on King’s B medium,"ncharacterized, identified and tested their potential in utilizing diesel, a petroleum hydrocarbon. At the same time, production"nof protease and urease enzymes during the utilization of diesel was also assayed following the standard procedures."nResults: The isolates were grown well on diesel and subsequently produced the extracellular enzymes protease and urease"nat significant levels when compared to their production in the absence of diesel. Optimum temperature and pH for increased"ngrowth by four isolates was found to be 37oC and pH 8.0 indicating the maximum utilization of diesel. All the isolates showed"nmaximum growth in medium with 100% diesel than 100% glycerol as carbon source, when tested with different proportions"nof diesel and glycerol as carbon sources. Plasmid profile of the isolates revealed that, all four Pseudomonas isolates harbored"ntwo low molecular weight plasmids; one with 3 Kb size and the other with 10 kb to 12 Kb size."nConclusion: The four Pseudomonas isolates of the present study were found to have potential in diesel degradation and can"nbe recommended for bioremediation of sites that are contaminated with diesel.
Carter, Gerald G.; Logsdon, Ryane; Arnold, Bryan D.; Menchaca, Angelica; Medellin, Rodrigo A.
Background Bat pups produce individually distinct isolation calls to facilitate maternal recognition. Increasing evidence suggests that, in group-living bat species, adults often use similar calls to maintain contact. We investigated if isolated adults from all three species of the highly cooperative vampire bats (Phyllostomidae: Desmodontinae) would produce vocally distinct contact calls when physically isolated. Methods/Principal Findings We assessed variation in contact calls recorded from isolated captive and wild-caught adult common vampire bats (Desmodus rotundus), white-winged vampire bats (Diaemus youngi) and hairy-legged vampire bats (Diphylla ecaudata). We compared species-typical contact call structure, and used information theory and permuted discriminate function analyses to examine call structure variation, and to determine if the individuality of contact calls is encoded by different call features across species and populations. We found that isolated adult vampire bats produce contact calls that vary by species, population, colony, and individual. However, much variation occurred within a single context and individual. We estimated signature information for captive Diaemus (same colony), captive Desmodus (same colony), and wild Desmodus (different colonies) at 3.21, 3.26, and 3.88 bits, respectively. Contact calls from a captive colony of Desmodus were less individually distinct than calls from wild-caught Desmodus from different colonies. Both the degree of individuality and parameters encoding individuality differed between the bats from a single captive colony and the wild-caught individuals from different groups. This result is consistent with, but not sufficient evidence of, vocal convergence in groups. Conclusion Our results show that adult vampire bats of all three species produce highly variable contact calls when isolated. Contact calls contain sufficient information for vocal discrimination, but also possess more intra-individual variation
Gerald G Carter
Full Text Available BACKGROUND: Bat pups produce individually distinct isolation calls to facilitate maternal recognition. Increasing evidence suggests that, in group-living bat species, adults often use similar calls to maintain contact. We investigated if isolated adults from all three species of the highly cooperative vampire bats (Phyllostomidae: Desmodontinae would produce vocally distinct contact calls when physically isolated. METHODS/PRINCIPAL FINDINGS: We assessed variation in contact calls recorded from isolated captive and wild-caught adult common vampire bats (Desmodus rotundus, white-winged vampire bats (Diaemus youngi and hairy-legged vampire bats (Diphylla ecaudata. We compared species-typical contact call structure, and used information theory and permuted discriminate function analyses to examine call structure variation, and to determine if the individuality of contact calls is encoded by different call features across species and populations. We found that isolated adult vampire bats produce contact calls that vary by species, population, colony, and individual. However, much variation occurred within a single context and individual. We estimated signature information for captive Diaemus (same colony, captive Desmodus (same colony, and wild Desmodus (different colonies at 3.21, 3.26, and 3.88 bits, respectively. Contact calls from a captive colony of Desmodus were less individually distinct than calls from wild-caught Desmodus from different colonies. Both the degree of individuality and parameters encoding individuality differed between the bats from a single captive colony and the wild-caught individuals from different groups. This result is consistent with, but not sufficient evidence of, vocal convergence in groups. CONCLUSION: Our results show that adult vampire bats of all three species produce highly variable contact calls when isolated. Contact calls contain sufficient information for vocal discrimination, but also possess more intra
Ser, Hooi-Leng; Palanisamy, Uma Devi; Yin, Wai-Fong; Chan, Kok-Gan; Goh, Bey-Hing; Lee, Learn-Han
Actinobacteria from the unique intertidal ecosystem of the mangroves are known to produce novel, bioactive secondary metabolites. A novel strain known as MUSC 136(T) (=DSM 100712(T) = MCCC 1K01246(T)) which was isolated from Malaysian mangrove forest soil has proven to be no exception. Assessed by a polyphasic approach, its taxonomy showed a range of phylogenetic and chemotaxonomic properties consistent with the genus of Streptomyces. Phylogenetically, highest similarity was to Streptomyces misionensis NBRC 13063(T) (99.6%) along with two other strains (>98.9% sequence similarities). The DNA-DNA relatedness between MUSC 136(T) and these type strains ranged from 22.7 ± 0.5% to 46.5 ± 0.2%. Overall, polyphasic approach studies indicated this strain represents a novel species, for which the name Streptomyces malaysiense sp. nov. is proposed. The potential bioactivities of this strain were explored by means of antioxidant and cytotoxic assays. Intriguingly, MUSC 136(T) exhibited strong antioxidative activities as evaluated by a panel of antioxidant assays. It was also found to possess high cytotoxic effect against HCT-116 cells, which probably mediated through altering p53 protein and intracellular glutathione levels. Chemical analysis of the extract using GC-MS further affirms that the strain produces chemopreventive related metabolites.
Sultan, Suandi Pratama; Kitani, Shigeru; Miyamoto, Kiyoko T; Iguchi, Hiroyuki; Atago, Tokitaka; Ikeda, Haruo; Nihira, Takuya
Streptomyces hormones, sometimes called as autoregulators, are important signaling molecules to trigger secondary metabolism across many Streptomyces species. We recently identified a butenolide-type autoregulator (termed avenolide) as a new class of Streptomyces hormone from Streptomyces avermitilis that produces important anthelmintic agent avermectin. Avenolide triggers the production of avermectin with minimum effective concentration of nanomolar. Here, we describe the characterization of avaR1 encoding an avenolide receptor in the regulation of avermectin production and avenolide biosynthesis. The disruption of avaR1 resulted in transcriptional derepression of avenolide biosynthetic gene with an increase in avenolide production, with no change in the avermectin production profile. Moreover, the avaR1 mutant showed increased transcription of avaR1. Together with clear DNA-binding capacity of AvaR1 toward avaR1 upstream region, it suggests that AvaR1 negatively controls the expression of avaR1 through the direct binding to the promoter region of avaR1. These findings revealed that the avenolide receptor AvaR1 functions as a transcriptional repressor for avenolide biosynthesis and its own synthesis.
Full Text Available From March 1999 to March 2000, we conducted a prospective multicenter study of candidemia involving five tertiary care hospitals from four countries in Latin America. Yeast isolates were identified by classical methods and the antifungal susceptibility profile was determined according to the National Committee for Clinical Laboratory Standards microbroth assay method. During a 12 month-period we were able to collect a total of 103 bloodstream isolates of Candida spp. C. albicans was the most frequently isolated species accounting for 42% of all isolates. Non-albicans Candida species strains accounted for 58% of all episodes of candidemia and were mostly represented by C. tropicalis (24.2% and C. parapsilosis (21.3%. It is noteworthy that we were able to identify two cases of C. lusitaniae from different institutions. In our casuistic, non-albicans Candida species isolates related to candidemic episodes were susceptible to fluconazole. Continuously surveillance programs are needed in order to identify possible changes in the species distribution and antifungal susceptibility patterns of yeasts that may occurs after increasing the use of azoles in Latin American hospitals.
Fernando José Vicenzi
Full Text Available Cystic fibrosis (CF patients with Burkholderia cepacia complex (Bcc pulmonary infections have high morbidity and mortality. The aim of this study was to compare different methods for identification of Bcc species isolated from paediatric CF patients. Oropharyngeal swabs from children with CF were used to obtain isolates of Bcc samples to evaluate six different tests for strain identification. Conventional (CPT and automatised (APT phenotypic tests, polymerase chain reaction (PCR-recA, restriction fragment length polymorphism-recA, recAsequencing, and matrix-assisted laser desorption ionization-time of flight (MALDI-TOF were applied. Bacterial isolates were also tested for antimicrobial susceptibility. PCR-recA analysis showed that 36 out of the 54 isolates were Bcc. Kappa index data indicated almost perfect agreement between CPT and APT, CPT and PCR-recA, and APT and PCR-recA to identify Bcc, and MALDI-TOF and recAsequencing to identify Bcc species. The recAsequencing data and the MALDI-TOF data agreed in 97.2% of the isolates. Based on recA sequencing, the most common species identified were Burkholderia cenocepacia IIIA (33.4%,Burkholderia vietnamiensis (30.6%, B. cenocepaciaIIIB (27.8%, Burkholderia multivorans (5.5%, and B. cepacia (2.7%. MALDI-TOF proved to be a useful tool for identification of Bcc species obtained from CF patients, although it was not able to identify B. cenocepacia subtypes.
WANG Zifeng; XIAO Tian; PANG Shaojun; LIU Min; YUE Haidong
We conducted this study to assess the diversity of bacteria associated with the surfaces of algae based on 16S rDNA sequence analyses. Twelve strains of bacteria were obtained from the surfaces of the following four species of algae: Gracilaria textorii, Ulva pertusa, Laminaria japonica, and Polysiphonia urceolata. The isolated strains of bacteria can be divided into two groups: Halomonas and Vibrio, in physiology, biochemical characteristics and 16S rDNA sequence analyses. The phylogenetic tree constructed based on 16S rDNA sequences of the isolates shows four obvious clusters, Halomonas venusta, Vibrio tasmaniensis, Vibrio lentus, and Vibrio splendidus. Isolates from the surface of P. Urceolata are more abundant and diverse, of which strains P9 and P28 have a 16S rDNA sequence very similar (97.5%-99.8%) to that of V. Splendidus. On the contrary, the isolates from the surfaces of G. Textorii, U. Pertusa and L. Japonica are quite simple and distribute on different branches of the phylogenetic tree. In overall, the results of this study indicate that the genetic relationships among the isolates are quite close and display a certain level of host species specificity, and alga-associated bacteria species are algal species specific.
da Silva, Krisle; Florentino, Ligiane Aparecida; da Silva, Karina Barroso; de Brandt, Evie; Vandamme, Peter; de Souza Moreira, Fatima Maria
The aim of the present study was to identify a collection of 35 Cupriavidus isolates at the species level and to examine their capacity to nodulate and fix N(2). These isolates were previously obtained from the root nodules of two promiscuous trap species, Phaseolus vulgaris and Leucaena leucocephala, inoculated with soil samples collected near Sesbania virgata plants growing in Minas Gerais (Brazil) pastures. Phenotypic and genotypic methods applied for this study were SDS-PAGE of whole-cell proteins, and 16S rRNA and gyrB gene sequencing. To confirm the ability to nodulate and fix N(2), the presence of the nodC and nifH genes was also determined, and an experiment was carried out with two representative isolates in order to authenticate them as legume nodule symbionts. All 35 isolates belonged to the betaproteobacterium Cupriavidus necator, they possessed the nodC and nifH genes, and two representative isolates were able to nodulate five different promiscuous legume species: Mimosa caesalpiniaefolia, L. leucocephala, Macroptilium atropurpureum, P. vulgaris and Vigna unguiculata. This is the first study to demonstrate that C. necator can nodulate legume species. Copyright Â© 2012 Elsevier GmbH. All rights reserved.
Vicenzi, Fernando José; Pillonetto, Marcelo; de Souza, Helena Aguilar Peres Homem de Mello; Palmeiro, Jussara Kasuko; Riedi, Carlos Antônio; Rosario-Filho, Nelson Augusto; Dalla-Costa, Libera Maria
Cystic fibrosis (CF) patients with Burkholderia cepacia complex (Bcc) pulmonary infections have high morbidity and mortality. The aim of this study was to compare different methods for identification of Bcc species isolated from paediatric CF patients. Oropharyngeal swabs from children with CF were used to obtain isolates of Bcc samples to evaluate six different tests for strain identification. Conventional (CPT) and automatised (APT) phenotypic tests, polymerase chain reaction (PCR)-recA, restriction fragment length polymorphism-recA, recAsequencing, and matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) were applied. Bacterial isolates were also tested for antimicrobial susceptibility. PCR-recA analysis showed that 36 out of the 54 isolates were Bcc. Kappa index data indicated almost perfect agreement between CPT and APT, CPT and PCR-recA, and APT and PCR-recA to identify Bcc, and MALDI-TOF and recAsequencing to identify Bcc species. The recAsequencing data and the MALDI-TOF data agreed in 97.2% of the isolates. Based on recA sequencing, the most common species identified were Burkholderia cenocepacia IIIA (33.4%),Burkholderia vietnamiensis (30.6%), B. cenocepaciaIIIB (27.8%), Burkholderia multivorans (5.5%), and B. cepacia (2.7%). MALDI-TOF proved to be a useful tool for identification of Bcc species obtained from CF patients, although it was not able to identify B. cenocepacia subtypes. PMID:26814642
Full Text Available Context: Candida species are opportunistic yeasts that cause infections ranging from simple dermatosis to potentially life-threatening fungemia. The emergence of resistance to antifungal drugs has been increased in the past two decades. Aim: the present study we determined to find out the susceptibility profiles of clinical isolates of Candida species against four antifungal drugs, including amphotericin B, ketoconazole, fluconazole and itraconazole. Materials and Methods: Antifungal susceptibility testing of the yeasts was done in accordance with the proposed guidelines for antifungal disk diffusion susceptibility testing of yeasts based on the CLSI document M44-A. Results: A total of 206 yeast isolates were assessed. Among the evaluated Candida species, the highest rates of resistance to ketoconazole were seen in Candida glabrata (16.6% and Candida albicans (3.2%. Susceptibility and intermediate response to fluconazole were seen in 96.6% and 3.4% of the Candida isolates, respectively. A total of 19 (9.2% yeast isolates showed petite phenomenon including 11 C. glabrata, 3 C. albicans, 2 Candida dubliniensis and one isolate of each Candida krusei and Candida parapsilosis. Conclusion: The high number of petite mutation in the isolated yeasts should be seriously considered since it may be one of the reasons of antifungal treatment failure.
Full Text Available The study was carried out to compare the efficacy of three media namely Modified Thayer Martin medium, McClung′s carbon free broth with paraffin bait and paraffin agar in isolating Nocardia species from clinical specimens. Two hundred and seventy six clinical specimens from 245 cases were studied which included cases of bronchopulmonary and systemic infections and cases of mycetoma. Paraffin agar was found to be an inexpensive and selective medium for isolation of Nocardia species when compared with Modified Thayer Martin medium and paraffin bait techniques.
Olumide A. Odeyemi; Ahmad, Asmat
This study aimed to investigate antibiotics resistance pattern and phenotyping of Aeromonas species isolated from different aquatic sources in Melaka, Malaysia. A total of 53 Aeromonas species were isolated from the following sources: sediment (n = 13), bivalve (n = 10), sea cucumber (n = 16) and sea water (n = 14) and resistance to 12 antibiotics – Tetracycline (30 μg), Kanamycin (30 μg), Oxytetracycline (30 μg), Ampicillin (10 μg), Streptomycin (10 μg), Gentamicin (10 μg), Sulphamethoxazole...
Olumide A. Odeyemi; Asmat Ahmad
This study aimed to investigate antibiotics resistance pattern and phenotyping of Aeromonas species isolated from different aquatic sources in Melaka, Malaysia. A total of 53 Aeromonas species were isolated from the following sources: sediment (n = 13), bivalve (n = 10), sea cucumber (n = 16) and sea water (n = 14) and resistance to 12 antibiotics – Tetracycline (30 μg), Kanamycin (30 μg), Oxytetracycline (30 μg), Ampicillin (10 μg), Streptomycin (10 μg), Gentamicin (10 μg), Sulphamethoxazole...
Lorusso, Alessio; Teodori, Liana; Leone, Alessandra; Marcacci, Maurilia; Mangone, Iolanda; Orsini, Massimiliano; Capobianco-Dondona, Andrea; Camma', Cesare; Monaco, Federica; Savini, Giovanni
A novel member of the Pteropine Orthoreovirus species has been isolated and sequenced for the whole genome from flying foxes (Pteropus vampyrus) imported to Italy from Indonesia. The new isolate named Indonesia/2010 is genetically similar to Melaka virus which has been the first virus of this species to be shown to be responsible for human respiratory disease. Our findings highlight the importance of flying foxes as vectors of potentially zoonotic viruses and the biological hazard that lies in the import of animals from geographical areas that are ecologically diverse from Europe.
Vorimore, Fabien; Hsia, Ru-Ching; Huot-Creasy, Heather; Bastian, Suzanne; Deruyter, Lucie; Passet, Anne; Sachse, Konrad; Bavoil, Patrik; Myers, Garry; Laroucau, Karine
Investigations conducted on feral African Sacred Ibises (Threskiornisaethiopicus) in western France led to the isolation of a strain with chlamydial genetic determinants. Ultrastructural analysis, comparative sequence analysis of the 16S rRNA gene, ompA, and of a concatenate of 31 highly conserved genes, as well as determination of the whole genome sequence confirmed the relatedness of the new isolate to members of the Chlamydiaceae, while, at the same time demonstrating a unique position outside the currently recognized species of this family. We propose to name this new chlamydial species Chlamydiaibidis .
Full Text Available Objective: This study evaluated the occurrence and antimicrobial susceptibility profile of Salmonella species isolated from various poultry products including chicken meat, poultry eggs, poultry bird's drinking water, and poultry feed. Materials and methods: A total of 79 samples comprising of chicken meat (n=20, egg shell (n=15, poultry egg contents (n=18, drinking water (n=14, and poultry feed (n=12 were bacteriologically and microscopically analyzed for the isolation of Salmonella species. Results: Overall, this study reported a high prevalence of Salmonella species (62% from various poultry products especially in poultry (chicken meat and poultry egg contents where the percentage occurrence of Salmonella species was 100% and 20.4% respectively. The antibiogram conducted on the Salmonella species isolated from the various poultry samples reveal that all the isolates were multi-drug resistant to more than 50% of the tested antibiotics especially to tetracycline, gentamicin, tobramycin, nitrofurantoin and imipenem. However, most of the Salmonella species were also found to be highly susceptible to ceftriaxone, cefotaxime, ertapenem and ceftazidime. It was also observed in this study that the highest level of resistance to the tested antibiotics was recorded in Salmonella species isolated from poultry meat samples. Conclusion: Salmonellosis due to the consumption of contaminated or infected poultry products could pose serious public health problem to the general public if allowed. Thus, poultry farms and other poultry product outlets should be operated under sanitized conditions that ward-off the incidence of foodborne pathogens such as Salmonella. The use of antibiotics as growth promoting agents and prophylaxis in the production of poultry birds in this region should be discouraged and ndash; since such practices allowed drug-resistant bacteria to emerge and spread in the community. [J Adv Vet Anim Res 2016; 3(4.000: 353-359
Irfan Ahmad Mir
Conclusions: Occurrence of high proportion of serovars in our study which can cause serious gastroenteritis in humans is a matter of concern. Salmonella Altona has been detected for the first time in India from poultry. This serotype is known to cause serious outbreaks of gastroenteritis in humans. Multidrug resistant isolates were recovered at high percentage which can be attributed to non-judicious use of antibiotics both in prophylaxis and treatment regimen. This observation draws serious attention as poultry serves as an important source of transmission of these multidrug resistant Salmonella serovars to humans.
de Folter Stefan
Full Text Available Abstract Background Small RNAs emerged over the last decade as key regulators in diverse biological processes in eukaryotic organisms. To identify and study small RNAs, good and efficient protocols are necessary to isolate them, which sometimes may be challenging due to the composition of specific tissues of certain plant species. Here we describe a simple and efficient method to isolate small RNAs from different plant species. Results We developed a simple and efficient method to isolate small RNAs from different plant species by first comparing different total RNA extraction protocols, followed by streamlining the best one, finally resulting in a small RNA extraction method that has no need of first total RNA extraction and is not based on the commercially available TRIzol® Reagent or columns. This small RNA extraction method not only works well for plant tissues with high polysaccharide content, like cactus, agave, banana, and tomato, but also for plant species like Arabidopsis or tobacco. Furthermore, the obtained small RNA samples were successfully used in northern blot assays. Conclusion Here we provide a simple and efficient method to isolate small RNAs from different plant species, such as cactus, agave, banana, tomato, Arabidopsis, and tobacco, and the small RNAs from this simplified and low cost method is suitable for downstream handling like northern blot assays.
Li, Xingpeng; Kerrigan, Julia; Chai, Wenxuan; Schnabel, Guido
Blackberry fruits symptomatic for gray mold were collected from three commercial blackberry fields in northwestern South Carolina. Single-spore isolates were generated and two distinct phenotypes were discovered in each location; one sporulated on PDA and one did not. One isolate of each phenotype and location (six isolates total) were selected for in depth molecular and morphological characterization. Glyceraldehyde-3-phosphate dehydrogenase (G3PDH), heat-shock protein 60 (HSP60) and DNA-dependent RNA polymerase subunit II (RPB2) coding sequence alignment revealed Botrytis cinerea as the sporulating phenotype and a new yet undescribed species as the non-sporulating phenotype. The new Botrytis sp., described herein as Botrytis caroliniana, was most closely related genetically to B. fabiopsis and B. galanthina, the causal agents of gray mold disease of broad bean and snowdrop, respectively. It produces smaller conidia than either B. fabiopsis or B. galanthina, and sequence analysis of genes encoding necrosis and ethylene-inducing proteins (NEPs) also indicated that the Botrytis isolates represent a separate and distinct species. The new species is pathogenic on blackberry fruits and broad bean leaves, which distinguishes it further from B. galanthina. The new species formed white to pale gray colonies with short, tufted aerial mycelium and produced black sclerotia on PDA at 20 C. To our knowledge this is only the third Botrytis species discovered to cause disease on blackberry in the United States.
The results of analysis of DNA-DNA hybridization confirmed that these isolates constitute a new and differentiated cluster within the genus Sinobacterium. The predominant fatty acids were C16: 0, C16: 1 ω7c/C16: 1 ω6c and C18: 1 ω7c. The G + C content of the new isolates is 52.2 ± 1 mol%. Based on phenotypic and phylogenetic characteristics, the seven isolates represent a new species for which the name Sinobacterium norvegicum sp. nov. is proposed, with strain 3CM4T (= CECT 8267T = CAIM 1884T as the type strain.
Morais, Camila G; Lara, Carla A; Oliveira, Evelyn S; Peter, Gábor; Dlauchy, Dénes; Rosa, Carlos A
Three strains of a new xylanase-producing yeast species were isolated from rotting wood samples collected in the Atlantic Rain Forest of Brazil. The sequences of the ITS region and D1/D2 domains of the large subunit of the rRNA gene showed that this new yeast species belongs to the genus Spencermartinsiella, and its closest relatives among the recognized species are S. europaea and S. ligniputridi. The novel species Spencermartinsiella silvicola sp. nov. is proposed to accommodate these isolates. The type strain is UFMG-CM-Y274T (= CBS 13490T). The MycoBank number is MB 813053. In addition, Candida cellulosicola is reassigned to the genus Spencermartinsiella as a new combination.
James, Stephen A; Cadet, Geneviève M; Barriga, Enrique Javier Carvajal; Barahona, Patricia Portero; Cross, Kathryn; Bond, Christopher J; Roberts, Ian N
A single strain, CLQCA-10-114(T), representing a novel yeast species belonging to the genus Saturnispora was isolated from the fruit of an unidentified species of bramble (Rubus sp.), collected from the Maquipucuna cloud forest reserve, near Quito, in Ecuador. Sequence analyses of the D1/D2 domains of the large-subunit rRNA gene and ribosomal internal transcribed spacer region indicated that the novel species is most closely related to the recently described species Saturnispora gosingensis, isolated from the fruiting body of a mushroom collected in Taiwan, and Saturnispora hagleri, a Drosophila-associated yeast found in Brazil. The name Saturnispora quitensis sp. nov. is proposed to accommodate this strain; the type strain is CLQCA-10-114(T) (=CBS 12184(T)=NCYC 3744(T)).
Mori, M; Ohta, M; Agata, N; Kido, N; Arakawa, Y; Ito, H; Komatsu, T; Kato, N
In the 77 reference strains for Klebsiella K types, there are 17 strains (22.1%) of Klebsiella planticola, 6 strains (7.8%) of Klebsiella oxytoca, 1 strain (1.3%) of Klebsiella terrigena, and 53 strains (68.8%) of Klebsiella pneumoniae. The species K. planticola, which was originally isolated from botanical and aquatic environments and hence thus named, was also identified at high incidence (81 strains, 18.5%) among the 439 recent clinical isolates of Klebsiella species. Among these K. planticola strains of hospital origin, 52 (64%) were isolated from sputum, 17 (21%) from urine, and the remaining 12 (15%) from other sources. The capsular types of these isolates were determined by the gel precipitation reaction. Seventy of 81 K. planticola isolates (86.4%) were typable by antisera to Klebsiella reference strains for K types and the K types of the clinical isolates distributed to 35 kinds of K types. The proportion of typable strains among clinical isolates of K. planticola was very similar to those in K. pneumoniae (87.5%) and K. oxytoca (86.0%).
Magalhães, Yankee C; Bomfim, Maria Rosa Q; Melônio, Luciane C; Ribeiro, Patrícia C S; Cosme, Lécia M; Rhoden, Cristianne R; Marques, Sirlei G
In this study, we isolated and phenotypically identified 108 yeast strains from various clinical specimens collected from 100 hospitalized patients at three tertiary hospitals in São Luís-Maranhão, Brazil, from July to December 2010. The isolates were analyzed for their susceptibility to four of the most widely used antifungal agents in the surveyed hospitals, amphotericin B, fluconazole, 5-flucytosine and voriconazole. The species identified were Candida albicans (41.4%), Candida tropicalis (30.1%), C. glabrata (7.4%), Candida parapsilosis (5.5%), Candida krusei (4.6%), Cryptococcus neoformans (4.6%), Trichosporon spp . (3.7%), Candida norvegensis (0.9%), Rhodotorula glutinis (0.9%) and Pichia farinosa (0.9%). A higher isolation rate was observed in the following clinical specimens: urine (54 isolates; 50%), respiratory tract samples (21 isolates; 19.4%) and blood (20 isolates; 18.6%). Candida albicans isolates were 100% sensitive to all antifungal agents tested, whereas Candida krusei and Crytococcus neoformans displayed intermediate resistance to 5-flucytosine, with Minimal Inhibitory Concentration (MIC) values of 8 mg/mL and 16 mg/mL, respectively. Both strains were also S-DD to fluconazole with an MIC of 16 mg/mL. C. tropicalis was resistant to 5-flucytosine with an MIC of 32 μg/mL. This study demonstrates the importance of identifying the yeast species involved in community and nosocomial infections.
Siitonen, Vilja; Räty, Kaj; Metsä-Ketelä, Mikko
The "'Streptomyces' genetics and secondary metabolism" laboratory course gives an introduction to the versatile soil dwelling Gram-positive bacteria "Streptomyces" and their secondary metabolism. The course combines genetic modification of "Streptomyces"; growing of the strain and protoplast preparation, plasmid…
Results: In present study, from a total of 118 isolates of lactobacilli were determined. Lactobacillus plantarum (24%, Lactobacillus casei (20% and Lactobacillus agillis (18% from facultative heterofermentative Lactobacilli and Lactobacillus delbrueckii (21%, Lactobacillus helveticus (14% and Lactobacillus salvariu s (3% from obligative homofermentative Lactobacilli were found to be more dominant species.Conclusions: So for achievement to organoleptic characteristics of traditional cheeses in industrial productions, mixed starters including dominant Lactobacillus species identified in cheeses can be employed.
Paula Matos Oliveira
Full Text Available BACKGROUND: Vulvovaginal candidiasis (VVC is the second most common vaginal infection. HIV-infection is a risk factor for this infection. OBJECTIVE: To determine the frequency of VVC and to describe the main Candida species isolated and their susceptibility to antifungal drugs in HIV-infected patients, compared to HIV-uninfected women in Salvador, Brazil. METHODS: Cross-sectional study including a group of 64 HIV-infected women and 76 uninfected women, followed up at the AIDS reference center and at the Gynecological Clinic of Escola Bahiana de Medicina e Saúde Pública (Salvador, Bahia, Brazil. RESULTS: Frequency of Candida spp. was higher in HIV-infected women (29.7% than in HIV-uninfected controls (14.5% (p = 0.02. The odds ratio value for vulvovaginal candidiasis in HIV-infected patients was 2.6 (95% CI: 1.07 - 6.32 p = 0.03. Candida albicans was the most commonly isolated species in both HIV-infected (52.3% and uninfected women (85.7%, followed by C. parapsolis in 17.6% and 14.3%, respectively. In HIV-infected women, C. glabrata, C. parapsilosis, and a coinfection of C. albicans and C. glabrata were also identified. There was no significant difference between Candida species isolated from the vaginal mucosa of women with VVC and colonization of the vaginal mucosa of HIV-infected and HIV-uninfected women. One C. glabrata isolate from an HIV-infected patient was resistant to fluconazole and other two isolates exhibited a dose-dependent susceptibility. CONCLUSION: Our results confirm a higher frequency of Candida spp. isolated from the vaginal mucosa of HIV-infected women and a broader spectrum of species involved. Only Candida glabrata isolates showed decreased susceptibility to fluconazole.
Iweriebor, Benson C; Obi, Larry C; Okoh, Anthony I
The genus Enterococcus is known to possess the capacity to acquire and disseminate antimicrobial resistant determinants alongside the ability to produce various virulence genes that enables it to establish infections. We assessed the prevalence and antibiogram profiles of Enterococcus spp. in faecal samples of dairy cattle. Faecal swab samples were collected from 400 dairy cattle from two commercial cattle farms in two rural communities in the Eastern Cape, South Africa. Confirmation of enterococci isolates was carried out by PCR targeting of the tuf gene. Species delineation was by species-specific primers targeting the superoxide dismutase (sod A) gene in a multiplex PCR assay. Isolates were screened for the presence of the following virulence genes (ace, gel E, esp, efa A, cyl A and hyl E) and antimicrobial resistance determinants to erythromycin, vancomycin and streptomycin were evaluated molecularly. A total of 340 isolates were confirmed as belonging to the genus Enterococcus . Species distribution among the isolates consisted of Enterococcus faecium (52.94 %) and Enterococcus durans (23.53 %) in preponderance compared to the three other species, namely Enterococcus faecalis (8.8 %), Enterococcus hirae (8.6 %) and Enterococcus casseliflavus (5.9 %). All were resistant to vancomycin, while 99 % showed resistance to aminoglycoside and 94 % to macrolide. Three virulence genes (ace, gel E and esp) were detected in almost all the confirmed isolates. The resistance determinants van B (19.7 %), van C1 (25 %), van C2/3 (26.3 %) erm B (40.29 %) and str A (50.88 %) were detected among the isolates. A high prevalence of multidrug-resistant enterococci isolates was detected in this study and the genetic repertoire to survive in the presence of antimicrobial agents was present in these organisms.
Paula Matos Oliveira
Full Text Available BACKGROUND: Vulvovaginal candidiasis (VVC is the second most common vaginal infection. HIV-infection is a risk factor for this infection. OBJECTIVE: To determine the frequency of VVC and to describe the main Candida species isolated and their susceptibility to antifungal drugs in HIV-infected patients, compared to HIV-uninfected women in Salvador, Brazil. METHODS: Cross-sectional study including a group of 64 HIV-infected women and 76 uninfected women, followed up at the AIDS reference center and at the Gynecological Clinic of Escola Bahiana de Medicina e Saúde Pública (Salvador, Bahia, Brazil. RESULTS: Frequency of Candida spp. was higher in HIV-infected women (29.7% than in HIV-uninfected controls (14.5% (p = 0.02. The odds ratio value for vulvovaginal candidiasis in HIV-infected patients was 2.6 (95% CI: 1.07 - 6.32 p = 0.03. Candida albicans was the most commonly isolated species in both HIV-infected (52.3% and uninfected women (85.7%, followed by C. parapsolis in 17.6% and 14.3%, respectively. In HIV-infected women, C. glabrata, C. parapsilosis, and a coinfection of C. albicans and C. glabrata were also identified. There was no significant difference between Candida species isolated from the vaginal mucosa of women with VVC and colonization of the vaginal mucosa of HIV-infected and HIV-uninfected women. One C. glabrata isolate from an HIV-infected patient was resistant to fluconazole and other two isolates exhibited a dose-dependent susceptibility. CONCLUSION: Our results confirm a higher frequency of Candida spp. isolated from the vaginal mucosa of HIV-infected women and a broader spectrum of species involved. Only Candida glabrata isolates showed decreased susceptibility to fluconazole.
Schmidt, E M; Pfennig, K S
A fundamental goal of biology is to understand how new species arise and are maintained. Female mate choice is potentially critical to the speciation process: mate choice can prevent hybridization and thereby generate reproductive isolation between potentially interbreeding groups. Yet, in systems where hybridization occurs, mate choice by hybrid females might also play a key role in reproductive isolation by affecting hybrid fitness and contributing to patterns of gene flow between species. We evaluated whether hybrid mate choice behaviour could serve as such an isolating mechanism using spadefoot toad hybrids of Spea multiplicata and Spea bombifrons. We assessed the mate preferences of female hybrid spadefoot toads for sterile hybrid males vs. pure-species males in two alternative habitat types in which spadefoots breed: deep or shallow water. We found that, in deep water, hybrid females preferred the calls of sterile hybrid males to those of S. multiplicata males. Thus, maladaptive hybrid mate preferences could serve as an isolating mechanism. However, in shallow water, the preference for hybrid male calls was not expressed. Moreover, hybrid females did not prefer hybrid calls to those of S. bombifrons in either environment. Because hybrid female mate choice was context-dependent, its efficacy as a reproductive isolating mechanism will depend on both the environment in which females choose their mates as well as the relative frequencies of males in a given population. Thus, reproductive isolation between species, as well as habitat specific patterns of gene flow between species, might depend critically on the nature of hybrid mate preferences and the way in which they vary across environments.
Yamaguchi, Sayaka; Sano, Ayako; Hiruma, Midori; Murata, Michiko; Kaneshima, Takashi; Murata, Yoshiteru; Takahashi, Hideo; Takahashi, Sana; Takahashi, Yoko; Chibana, Hiroji; Touyama, Hidemi; Ha, Nguyen Thi Thanh; Nakazato, Yasutomo; Uehara, You; Hirakawa, Morihiko; Imura, Yoshimi; Terashima, Yoshie; Kawamoto, Yasuhiro; Takahashi, Keji; Sugiyama, Kazutoshi; Hiruma, Masataro; Murakami, Masaru; Hosokawa, Atsushi; Uezato, Hiroshi
We investigated 793 bird combs [645 chickens and 148 fighting cocks (Shamo)] to determine the prevalence of dermatophytes and their related fungal species. The targeted fungal species were recovered from 195 of the 793 examined birds (24.6 %). Prevalence ratios were compared in temperate (the mainland) and subtropical (Nansei Islands) areas, genders, strains, breeding scale (individual and farm), and housing system (in cage and free ranging). The frequency of the fungal species in the mainland, males, fighting cocks, breeding scale by individual nursing, and free-range housing system exhibited significantly higher positive ratios than that in the other groups. A total of 224 dermatophytes and related species were isolated, including 101 Arthroderma (Ar.) multifidum, 83 Aphanoascus (Ap.) terreus, five Uncinocarpus queenslandicus, two U. reesii, two Ap. pinarensis, one Amauroascus kuehnii, one Ar. simii, one Gymnoascus petalosporus, one Microsporum gallinae, and 28 Chrysosporium-like (Chrysosporium spp.) isolates, which were identified using internal transcribed spacer regions of ribosomal RNA gene sequences. The predominant fungal species in the mainland was Ap. terreus and that in the Nansei Islands was Ar. multifidum. Pathogenic fungal species to humans and animals were limited to M. gallinae and Ar. simii, which corresponded to 0.025 % of the isolates in this study.
Medema, Marnix H.; Breitling, Rainer; Takano, Eriko
Actinomycete bacteria of the genus Streptomyces are major producers of bioactive compounds for the biotechnology industry. They are the source of most clinically used antibiotics, as well as of several widely used drugs against common diseases, including cancer . Genome sequencing has revealed that
Medema, Marnix H.; Breitling, Rainer; Takano, Eriko; Voigt, C
Actinomycete bacteria of the genus Streptomyces are major producers of bioactive compounds for the biotechnology industry. They are the source of most clinically used antibiotics, as well as of several widely used drugs against common diseases, including cancer. Genome sequencing has revealed that t
Jorjandi, M.; G. H.S. Bonjar; A. Baghizadeh; G. R. S. Sirchi; H. Massumi; F. Baniasadi; S. Aghighi; P. R. Farokhi
Problem statement: Soil actinomycetes particularly Streptomyces spp. showed antagonistic activity against wide range of plant pathogens. In the recent decades they have attracted high interests as biocontrol agents. Onion neck rot or gray mold caused by Botrytis allii have imposed economic post harvest damages to onion bulbs and decreased its storage durability and market value. Approach: To investigate for biocontrol means against the pathogen, antagonisti...
Mirajkar, Nandita S.; Davies, Peter R.
Outbreaks of swine dysentery, caused by Brachyspira hyodysenteriae and the recently discovered “Brachyspira hampsonii,” have reoccurred in North American swine herds since the late 2000s. Additionally, multiple Brachyspira species have been increasingly isolated by North American diagnostic laboratories. In Europe, the reliance on antimicrobial therapy for control of swine dysentery has been followed by reports of antimicrobial resistance over time. The objectives of our study were to determine the antimicrobial susceptibility trends of four Brachyspira species originating from U.S. swine herds and to investigate their associations with the bacterial species, genotypes, and epidemiological origins of the isolates. We evaluated the susceptibility of B. hyodysenteriae, B. hampsonii, Brachyspira pilosicoli, and Brachyspira murdochii to tiamulin, valnemulin, doxycycline, lincomycin, and tylosin by broth microdilution and that to carbadox by agar dilution. In general, Brachyspira species showed high susceptibility to tiamulin, valnemulin, and carbadox, heterogeneous susceptibility to doxycycline, and low susceptibility to lincomycin and tylosin. A trend of decreasing antimicrobial susceptibility by species was observed (B. hampsonii > B. hyodysenteriae > B. murdochii > B. pilosicoli). In general, Brachyspira isolates from the United States were more susceptible to these antimicrobials than were isolates from other countries. Decreased antimicrobial susceptibility was associated with the genotype, stage of production, and production system from which the isolate originated, which highlights the roles of biosecurity and husbandry in disease prevention and control. Finally, this study also highlights the urgent need for Clinical and Laboratory Standards Institute-approved clinical breakpoints for Brachyspira species, to facilitate informed therapeutic and control strategies. PMID:27252458
Full Text Available Actinomycetes isolated from marine sediments along the southeast coast of Bay of Bengal were investigated for amylolytic activity. Marine actinomycete BTSS 1001 producing an alkaline amylase was identified from marine sediment of Diviseema coast, Bay of Bengal. The isolate produced alkaline amylase with maximum amylolytic activity at pH 9.5 at 50°C. The organism produced white to pale grey substrate mycelium and grayish aerial mycelium with pinkish brown pigmentation. A comprehensive study of morphological, physiological parameters, cultural characteristics, and biochemical studies was performed. The presence of iso-C15 : 0, anteiso-C15 : 0, iso-C16 : 0, and anteiso-C17 : 0 as the major cellular fatty acids, LL-diaminopimelic acid as the characteristic cell wall component, and menaquinones MK-9H(6 and MK-9H(8 as the major isoprenoid quinones is attributed to the strain BTSS 1001 belonging to the genus Streptomyces. Comparison of 16S rRNA gene sequences showed that strain BTSS 1001 exhibited the highest similarities to the type strains of Streptomyces rochei (99%, Streptomyces plicatus (99%, and Streptomyces enissocaesilis (99%. Using the polyphasic taxonomical approach and phenotypic characteristic studies, the isolate BTSS 1001 was characterized as marine actinomycete Streptomyces rochei.
Vega-Sánchez, Vicente; Acosta-Dibarrat, Jorge; Vega-Castillo, Fernando; Castro-Escarpulli, Graciela; Aguilera-Arreola, Ma Guadalupe; Soriano-Vargas, Edgardo
In the present study, Aeromonas isolates from diseased and healthy farmed rainbow trout (Oncorhynchus mykiss) in Mexico, were characterized phenotypically and identified to species level by using 16S rDNA RFLP-PCR. A total of 50 isolates were included in the study and 10 Aeromonas species identified. The species A. veronii biovar sobria (22%), A. hydrophila (20%) and A. bestiarum (20%) were the most predominant. All isolates (100%) were resistant to cephalothin.
Sudha; S; Masilamani; Selvam; M
Objective:To investigate the cytotoxic activity of actinomycete isolated from marine sediment.Methods:In the present study the DNA was isolated and the ITS region of 16s rRNA was amplified by polymerase chain reaction,using two universal bacterial primers,1492K（5’-GGTTACCTTG’TTAC GACTT-3’）and Eubac27F（5’-AGAGTTTGATCCTGGCTC AG-3’）.The amplified products were purified using TIANgel mini purification kit,ligated to MD18-T simple vector（TaKaRa）,and transformed into competent cells of Escherichia coli DH5α.16S rRNA gene fragment was sequenced using forward primer M13F（-47）and reverse primer M13R（-48）.Blast search sequence similarity was found against the existing non-redundanl nucleotide sequence database thus,identified as Streptomyces sp SU,Streptomyces rubralavandulae strain SU1,Streptomyces cacaoi strain SU2,Streptomyces cavourensis strain SU3,Streptomyces avidinii strain SU4,Streptomyces globisporus strain SU5,Streptomyces variabilis strain SU6,Streptomyces coelicolor strain SU 7.Among the eight identified isolates,one actinomycete Streptomyces avidinii strain SU4 was selected for further study.Results:Crude extract of the actinomycete isolate exhibited IC50in 64.5μg against Hep-2 cell line,250μg in VERO cell line.This value is very close to the criteria of cytotoxicity activity for the crude extracts,as established by the American National Cancer Institute（NCI）is in IC50<30μg/mL.The CC MS analysis showed that the active principle might be 1,2-benzenedicarboxylic acid,bis（2-methylpropyl）ester（12.17%）,isooctyl phthalate（15.29%）with the retention time 15.642 and 21.612,respectively.Conclusions:This study clearly proves that the marine sediment derived actinomycetes with bioactive metabolites can be expected to provide high quality biological material for high throughout biochemical and anticancer screening programs.These results help us to conclude that the potential of using metabolic engineering and post genomic
Labeda, David P
Multi-locus sequence analysis has been demonstrated to be a useful tool for identification of Streptomyces species and was previously applied to phylogenetically differentiate the type strains of species pathogenic on potatoes (Solanum tuberosum L.). The ARS Culture Collection (NRRL) contains 43 strains identified as Streptomyces scabiei deposited at various times since the 1950s and these were subjected to multi-locus sequence analysis utilising partial sequences of the house-keeping genes atpD, gyrB, recA, rpoB and trpB. Phylogenetic analyses confirmed the identity of 17 of these strains as Streptomyces scabiei, 9 of the strains as the potato-pathogenic species Streptomyces europaeiscabiei and 6 strains as potentially new phytopathogenic species. Of the 16 other strains, 12 were identified as members of previously described non-pathogenic Streptomyces species while the remaining 4 strains may represent heretofore unrecognised non-pathogenic species. This study demonstrated the value of this technique for the relatively rapid, simple and sensitive molecular identification of Streptomyces strains held in culture collections.
Bae, Kyung Sook; Kim, Mi Sun; Lee, Ji Hee; Kang, Joo Won; Kim, Dae In; Lee, Ji Hee; Seong, Chi Nam
To understand the isolation and classification state of actinobacterial species with valid names for Korean indigenous isolates, isolation source, regional origin, and taxonomic affiliation of the isolates were studied. At the time of this writing, the phylum Actinobacteria consisted of only one class, Actinobacteria, including five subclasses, 10 orders, 56 families, and 330 genera. Moreover, new taxa of this phylum continue to be discovered. Korean actinobacterial species with a valid name has been reported from 1995 as Tsukamurella inchonensis isolated from a clinical specimen. In 1997, Streptomyces seoulensis was validated with the isolate from the natural Korean environment. Until Feb. 2016, 256 actinobacterial species with valid names originated from Korean territory were listed on LPSN. The species were affiliated with three subclasses (Acidimicrobidae, Actinobacteridae, and Rubrobacteridae), four orders (Acidimicrobiales, Actinomycetales, Bifidobacteriales, and Solirubrobacterales), 12 suborders, 36 families, and 93 genera. Most of the species belonged to the subclass Actinobacteridae, and almost of the members of this subclass were affiliated with the order Actinomycetales. A number of novel isolates belonged to the families Nocardioidaceae, Microbacteriaceae, Intrasporangiaceae, and Streptomycetaceae as well as the genera Nocardioides, Streptomyces, and Microbacterium. Twenty-six novel genera and one novel family, Motilibacteraceae, were created first with Korean indigenous isolates. Most of the Korean indigenous actionobacterial species were isolated from natural environments such as soil, seawater, tidal flat sediment, and fresh-water. A considerable number of species were isolated from artificial resources such as fermented foods, wastewater, compost, biofilm, and water-cooling systems or clinical specimens. Korean indigenous actinobacterial species were isolated from whole territory of Korea, and especially a large number of species were from Jeju
González, Gloria M; Casillas-Vega, Néstor; Garza-González, Elvira; Hernández-Bello, Romel; Rivera, Gildardo; Rodríguez, Jesús Ancer; Bocanegra-Garcia, Virgilio
Cryptococcosis is caused by members of the Cryptococcus neoformans/Cryptococcus gattii species complex. Based on molecular identification, these two species have been further differentiated into molecular types. The aim of this work was to characterize clinical cryptococcal isolates recovered from six hospitals in Northeast Mexico from 1995 to 2011. One hundred and sixty-six isolates, which were characterized by biochemical tests and in vitro susceptibility to amphotericin B, fluconazole, and voriconazole, and M13 PCR fingerprinting, were included in this study. Utilizing phenotypic tests, 153 isolates (92.16 %) were identified as C. neoformans and 13 (7.83 %) as C. gattii. All isolates were susceptible to all antifungals tested. Employing M13 PCR fingerprinting, eight molecular types were detected. VNI was the most common genotype (124 cases; 74.6 %), followed by VNII (15 cases; 9 %), VNIII (8 cases; 4.8 %), VNIV (6 cases; 3.6 %), VGI (6 cases; 3.6 %), VGII (3 cases; 1.8 %), and VGIII and VGIV (2 cases, 1.2 % each). We confirm the presence of C. gattii in clinical isolates in Northeast Mexico, and a high clonal diversity in the studied strains of C. neoformans/C. gattii species complex.
Igbinosa, Isoken H; Igbinosa, Etinosa O; Okoh, Anthony I
This study aimed to assess the antibiogram of Aeromonas strains recovered from cattle faeces and the potential pathogenic status of the isolates. The antibiogram of the Aeromonas isolates demonstrated total resistance to clindamycin oxacillin, trimethoprim, novobiocin and ticarcillin. However, Aeromonas strains were sensitive to cefotaxime, oxytetracycline and tobramycin. The Aeromonas strains from Lovedale and Fort Cox farms were found to possess some virulence genes. The percentage distribution was aer 71.4%, ast 35.7%, fla 60.7%, lip 35.7% and hlyA 25% for Lovedale farm and aer 63.1%, alt 10.5%, ast 55.2%, fla 78.9%, lip 21% and hlyA 35.9% for Fort Cox farm. Class 1 integron was present in 27% of Aeromonas isolates; the bla TEM gene was present in 34.8%, while the blaP1 class A β-lactamase gene was detected in 12.1% of the isolates. Approximately 86% of the isolates formed a biofilm on microtitre plates. The presence of multiple antibiotic resistance and virulence genes in Aeromonas isolates from cattle faeces reveals the pathogenic and infectious importance of these isolates and is of great significance to public health. The possession of a biofilm-forming capability by such isolates may lead to difficulty during the management of infection related to Aeromonas species.
Léon, Fabian; Van Boven, Maurits; de Witte, Peter; Busson, Roger; Cokelaere, Marnix
A mixture of lysophosphatidylcholine (LPC) and phosphatidylcholine (PC) has been isolated by column chromatography from a jojoba meal (Simmondsia chinensis) extract. The molecular species of both classes could be separated and isolated by C18 reversed phase HPLC. The two major compounds were identified by 1D and 2D (1)H and (13)C NMR, by MS, and by GC-MS as 1-oleoyl-3-lysophosphatidylcholine and 1,2-dioleoyl-3-phosphatidylcholine. Eight other molecular species of LPC and four other molecular species of PC could be assigned by comparison of the mass spectra of the isolated compounds with the spectra of the two major compounds. Complete characterization of the individual molecular species was achieved by GC and GC-MS analysis of the fatty acyl composition from the isolated compounds. The PC/LPC proportion in the phospholipid mixture from three different samples is 1.6 +/- 0.1. LPC is considered to be an important bioactive compound; the results of this study suggest further research for the evaluation of potential health benefits of jojoba meal phospholipids.
Full Text Available Herein, we report the main characteristics of a new species named Pacaella massiliensis gen. nov., sp. nov., strain Marseille-P2670T (CSUR P2670 that was isolated from the gut microbiota of a 45-year-old French patient.
Yilmaz, Neriman; López-Quintero, Carlos A.; Vasco-Palacios, Aída Marcela; Frisvad, Jens C.; Theelen, Bart; Boekhout, Teun; Samson, Robert A.; Houbraken, Jos
Various Talaromyces strains were isolated during a survey of fungi involved in leaf litter decomposition in tropical lowland forests in the Caquetá and Amacayacu areas of the Colombian Amazon. Four new Talaromyces species are described using a polyphasic approach, which includes phenotypic character
Full Text Available We report the summary of main characteristics of Tidjanibacter massiliensis strain Marseille-P3084T, a new bacterial species isolated from the liquid sample of the colon of a patient with a history of irritable bowel syndrome.
Schmid, G P; Steere, A C; Kornblatt, A N; Kaufmann, A F; Moss, C W; Johnson, R C; Hovind-Hougen, K; Brenner, D J
Leptospira strain 10, which represents a new Leptospira species, was isolated from a skin biopsy of a patient with Lyme disease. Although pathogenic for laboratory animals, the organism was not considered to have a significant role in the patient's illness. PMID:3760144
Full Text Available The most recent taxonomic revision in Trichinella genus included 10 taxa. Trichinella spiralis (Ts is a thoroughly studied species but most of the isolates came from the Europe or American continents. Few information is available from China about the Trichinella isolates and their diversity. In this report two Chinese isolates were characterized and compared with European strains. The in vitro release of newborn larvae (NBL was determined for the two species. NBL is observed in supernatant of cell culture after adults purification on day 4 post infection (pi with Ts, and on day 5 pi with Trichinella nativa (Tna. A new parameter was thus proposed to characterize Trichinella strains. Restriction fragment length polymorphism (RFLP of genomic DNA and random amplified polymorphic cDNA (RAPD were used to define genetic variability among different isolates. Species specific pattern could be obtained with RAPD but it was far more difficult to get geographical markers for Trichinella using these methods. A low genetic variability in Trichinella species (i.e a strong "clonality" is suggested.
Ottonelli Stopiglia, Cheila Denise; Magagnin, Cibele Massotti; Castrillón, Mauricio Ramírez; Mendes, Sandra Denise Camargo; Heidrich, Daiane; Valente, Patricia; Scroferneker, Maria Lúcia
Sporotrichosis is a subacute or chronic mycosis caused worldwide by the dimorphic species complex, Sporothrix schenckii. We studied 85 isolates recovered in Brazil to verify their identification and evaluate their in vitro antifungal susceptibility patterns. Based on phenotypic tests (microscopic features, ability to grow at 30°C and 37°C, colony diameters, as well as assimilation of sucrose and raffinose) and molecular assays (amplification of a fragment of the calmodulin gene), the strains were identified as S. schenckii, S. brasiliensis and S. globosa, with a predominance of S. schenckii isolates. There was 37.7% disagreement between the phenotypic and genotypic identification methodologies. In general, terbinafine was the most active drug, followed by ketoconazole and itraconazole, and the less active fluconazole and voriconazole. Five isolates (one S. globosa and four S. schenckii) were found to be itraconazole-resistant strains but, in general, there were no differences in the in vitro antifungal susceptibility profiles among the Sporothrix species.
Skall, Helle Frank; Mellergaard, Stig; Olesen, Niels Jørgen
and the Kattegat. Surveillance of aquacultured fish, mainly rainbow trout (Oncorhynchus mykiss), for VHS, IHN and IPN, in Denmark during the previous 30 years, have resulted in Birnavirus serogroup B isolations twice. These isolations were from healthy rainbow trout. Samples sent to the Danish Veterinary......During cruises with Danish research vessels more than 17,000 wild marine fish belonging to 41 different species were sampled and examined virologically. Birnavirus serogroup B was isolated from 7 marine fish species: plaice (Pleuronectes platessa), dab (Limanda limanda), flounder (Platichthys...... Laboratory for virological examination and characterisation have revealed the presence of Birnavirus serogroup B in haddock (ML Melanogrammus aeglefinus), plaice and dab from wild marine fish caught by Irish fishermen and from Icelandic farmed halibut (Hippoglossus hippoglossus)....
Full Text Available A study of 192 strains of Coagulase negative staphylococcus (CONS showed that Staphylococcus epidermidis was the most common species, 158 (82.29% isolated from all clinical specimens followed by S. saprophyticus (30, 15.62% isolated mainly from urine. Slime production was exhibited by 77 (48.7% strains of S. epidermidis and 8 (26.6% of S. saprophyticus and the difference in the slime producing activity was statistically significant (p<0.005. Antibiotic susceptibility testing against 15 commonly used antibiotics showed multidrug resistance with more than 90% resistance to penicillin, more than 50% to cephalexin and ciprofloxacin and more than 20% to methicillin, thus, highlighting the importance of species identification and antibiotic susceptibility testing for clinical isolates of CONS.
Naz, Tayyaba; Khan, Muhammad Daud; Ahmed, Iftikhar; Rehman, Shafiq Ur; Rha, Eui Shik; Malook, Ijaz; Jamil, Muhammad
Heavy metal-resistant bacteria can be efficient bioremediators of metals and may provide an alternative or additional method to conventional methods of metal removal. In this study, 10 bacterial isolates were isolated from soil samples of a sugar industry, located at Peshawar, Pakistan. Morphological, physiological, and biochemical characteristics of these isolates were observed. Sequence analysis (16S ribosomal RNA) revealed that isolated strains were closely related to the species belonging to the genera Pseudomonas, Arthrobacter, Exiguobacterium, Citrobacter, and Enterobacter Bacterial isolates were resistant with a minimum inhibitory concentration (500-900 ppm) to lead ion (Pb(2+)), (500-600 ppm) nickel ion (Ni(2+)), (500-800 ppm) copper ion (Cu(2+)), and (600-800 ppm) chromium ion (Cr(3+)) in solid media. Furthermore, biosorption of metals proved considerable removal of heavy metals by isolated metal-resistant strains. Pseudomonas sp. reduced 37% (Pb(2+)), 32% (Ni(2+)), 29% (Cu(2+)), and 32% (Cr(3+)) and was thus found to be most effective, whereas Enterobacter sp. reduced 19% (Pb(2+)), 7% (Ni(2+)), 14% (Cu(2+)), and 21% (Cr(3+)) and was found to be least effective. While average reduction of Pb(2+), Ni(2+), Cu(2+), and Cr(3+) by Citrobacter sp. was found to be 24%, 18%, 23%, and 27%, respectively, among recognized species. This study revealed that Pseudomonas sp. may provide a new microbial community that can be used for enhanced remediation of contaminated environment.
Holmberg, Mia; Rosendal, Thomas; Engvall, Eva O; Ohlson, Anna; Lindberg, Ann
The aims of this study were to investigate the prevalence of Campylobacter species in Swedish dogs, to identify the species of the Campylobacter isolates and to genotype the C. jejuni isolates. Young and healthy dogs were targeted and the sampling was performed at 11 veterinary clinics throughout Sweden from October 2011 to October 2012. Faecal swab samples were collected and sent to the laboratory at the National Veterinary Institute (SVA) for isolation of Campylobacter, speciation and genotyping. Campylobacter spp. were isolated from 67 of the 180 sampled dogs which yields an overall prevalence of 37%. The most prevalent species of Campylobacter among the participating dogs was C. upsaliensis with 52 of the 67 identified isolates. A lower prevalence was observed for C. jejuni with seven identified isolates and one isolate was identified as C. helveticus. Multi-locus sequence typing (MLST) was carried out on the seven C. jejuni isolates and all sequence types that were found are also commonly found in humans. The dogs were divided into three age groups; 1) under 12 months, 2) 12 to 23 months and 3) 24 months and older. The highest prevalence was found in the two younger age groups. Dogs shedding C. jejuni were between 3-12 months of age while dogs shedding C. upsaliensis were found in all ages. The present investigation finds that Campylobacter spp. known to cause campylobacteriosis in humans are present in Swedish dogs. The results suggest an age predisposition where dogs under 2 years of age are more likely to shed Campylobacter spp. than older dogs. The most commonly isolated species was C. upsaliensis followed by C. jejuni, which was only detected in dogs up to 12 months of age. All C. jejuni isolates identified in the present study were of the same MLST types that have previously been described both in humans and in animals. The awareness of the Campylobacter risk of healthy young dogs may be an important way to reduce the transmission from dogs to infants
Full Text Available Streptomyces sp. IK isolated from compost inoculants, could produce extra cellular chitinase in a medium containing 0.2% (w/v colloidal chitin, fermented for 96 hours at 30oC. The enzyme was purified by a combination of ammonium sulphate precipitation and DEAE-Cellulose anion-exchange chromatography. On SDS-polyacrylamide gel electrophoresis analysis, the purified enzyme showed a mass of 71 kDa. Chitinase was optimally active at pH of 6.7 and at 37oC. Km value and Vmax of the protein for colloidal chitin were 2.92 mg/ml and 4.26 ìg/h, respectively.Key words : chitinase, Streptomyces, purification, characterization
Mustafa, Abu S; Habibi, Nazima; Osman, Amr; Shaheed, Faraz; Khan, Mohd W
Brucellosis is a zoonotic disease of major concern in Kuwait and the Middle East. Human brucellosis can be caused by several Brucella species with varying degree of pathogenesis, and relapses are common after apparently successful therapy. The classical biochemical methods for identification of Brucella are time-consuming, cumbersome, and provide information limited to the species level only. In contrast, molecular methods are rapid and provide differentiation at intra-species level. In this study, four molecular methods [16S rRNA gene sequencing, real-time PCR, enterobacterial repetitive intergenic consensus (ERIC)-PCR and multilocus variable-number tandem-repeat analysis (MLVA)-8, MLVA-11 and MLVA-16 were evaluated for the identification and typing of 75 strains of Brucella isolated in Kuwait. 16S rRNA gene sequencing of all isolates showed 90-99% sequence identity with B. melitensis and real-time PCR with genus- and species- specific primers identified all isolates as B. melitensis. The results of ERIC-PCR suggested the existence of 75 ERIC genotypes of B. melitensis with a discriminatory index of 0.997. Cluster classification of these genotypes divided them into two clusters, A and B, diverging at ~25%. The maximum number of genotypes (n = 51) were found in cluster B5. MLVA-8 analysis identified all isolates as B. melitensis, and MLVA-8, MLVA-11 and MLVA-16 typing divided the isolates into 10, 32 and 71 MLVA types, respectively. Furthermore, the combined minimum spanning tree analysis demonstrated that, compared to MLVA types discovered all over the world, the Kuwaiti isolates were a distinct group of MLVA-11 and MLVA-16 types in the East Mediterranean Region.
Vaz Jauri, Patricia; Kinkel, Linda L
Chemical communication among kin bacteria modulates diverse activities. Despite the general consensus that signaling among non-kin organisms is likely to influence microbial behavior, there is limited information on the potential for microbial interactions to alter microbial phenotypes in natural habitats. We explored patterns of interaction that alter inhibitory phenotypes among Streptomyces isolates from distinct communities. Shifts in inhibition in response to the presence of a partner were evaluated for 861 isolate combinations, and were considered in relation to nutrient use, 16S sequence, inhibition phenotype and community origin. The frequency of inhibition-shifting interactions was significantly higher among isolates from the same (0.40) than from different (0.33) communities, suggesting local selection for inhibition-shifting interactions. Communities varied in the frequency with which Streptomyces isolates responded to a partner but not in the frequency with which isolates induced changes in partners. Streptomyces isolates were more likely to exhibit increased inhibition of a target bacterium in response to isolates that compete for the same nutrients, are closely-related or are strongly inhibited by their antibiotics. This work documents a high frequency of interactions among Streptomyces that shift the capacity of Streptomyces to inhibit other microbes, and suggests significant potential for such interactions to shape microbial community dynamics.
Trindade, Rita C; Resende, Maria A; Pimenta, Raphael S; Lachance, Marc-André; Rosa, Carlos A
Sixteen strains of the new yeast species Candida sergipensis have been isolated from frozen pulps of the tropical fruits umbú ( Spondias tuberosa Avr. Cam.) and mangaba ( Hancornia speciosa Gom.). Candida sergipensis was one of the prevalent species in the yeast community of these substrates. The new asexual ascomycetous yeast is phylogenetically related to Candida spandovensis and Candida sorbophila, species belonging to the Wickerhamiella clade, as evidenced by the sequences of the D1/D2 domains of their large subunit ribosomal DNAs. The species C. sergipensis and C. spandovensis can be separated on the basis of growth on 50% glucose agar, xylose and succinate, negative for the first species and positive for the second. The type culture is strain UFMG-R188 (CBS 9567).
Szakacs G; Nagy V; Kovacs K
@@ The relationship between taxonomic status of Trichoderma spp., chitinase production in solid substrate fermentation (SSF) on four media and mycoparasitism in dual culture (confrontation assay)against four plant pathogenic fungi was studied. Seventy five Trichoderma isolates belonging to 35species have been screened. The plant pathogenic fungi used in confrontation assay were Botrytis cinerea , Fusarium oxysporum f. sp. dianthi , Rhizoctonia solani and Sclerotinia sclerotiorum . The SSF media contained wheat bran, crude chitin (from crab shells, SIGMA) and salt solutions. The best performing isolates in mycoparasitism tests were Trichoderma flavofuscum, T. harzianum, T.inhamatum, T. koningii and T. strigosum. Some isolates exhibiting good mycoparasitism produced chitinase in SSF only at low or medium level. In contrary there were isolates with excellent extracellular chitinase production but their biocontrol potential did not belong to the leading group.Statistical methods have been used to evaluate the data.
Self-resistance has been investigated in Streptomyces caelestis (producer of the lincosamide antibiotic celesticetin), from which a lincosamide resistance determinant (clr) has been isolated on a 1-kilobase DNA fragment and cloned in Streptomyces lividans. The clr product is a specific methylase which produces a single residue of N6-monomethyladenine in 23S rRNA at position 2058, thereby rendering the 50S ribosmal subunit resistant to the action of lincosamides.
Calcutt, M J; Cundliffe, E
Self-resistance has been investigated in Streptomyces caelestis (producer of the lincosamide antibiotic celesticetin), from which a lincosamide resistance determinant (clr) has been isolated on a 1-kilobase DNA fragment and cloned in Streptomyces lividans. The clr product is a specific methylase which produces a single residue of N6-monomethyladenine in 23S rRNA at position 2058, thereby rendering the 50S ribosmal subunit resistant to the action of lincosamides. PMID:2376570
Kocić-Tanackov Sunčica D.
Full Text Available Fungi of the genus Fusarium, known as toxigenic species, are very of- ten parasites and contaminants of brewer's barley. In this paper, the composition of the genus Fusarium species in brewer's barley samples and their potential in the zearalenone synthesis were investigated. The tests were done on different brewer's barley varieties, crop 2003, samples (SSK1, SSK2, SSK3 SSK4, SSK5, SSK6, SSK7, SSK8, SSK9, SSK10 and SSK12 from Kragujevac locality. The isolation and identification of the Fusarium species were done according to the methods described by N e l s o n et al. (1983. The identified Fusarium species (6 were tested for their capacity to synthesise zearalenone. The isolates were cultivated on sterilised barley grains at the temperature of 25°C for 14 days, and then the zearalenone concentration was determined by the fluorometric method on the fluorometer "VI- CAM" series 4. The following seven Fusarium species were isolated from barley samples: F. acuminatum, F. avenaceum, F. culmorum, F. equiseti, F. poae, F. sporotrichioides and F. tricinctum. F. poae was the most distributed species (10.26%. The zearalenone concentration within the range of 12.0 to 430.0 g kg-1 was determined in cultures of barley grain inoculated with F. avenacuem (SSK6 and SSK12, F. culmorum (SSK8, F. tricinctum (SSK1, F. sporotrichioides (SSK7 and SSK12 and F. poae (SSK5, SSK9 and SSK10. Isolates of F. equiseti (SSK2 and F. poae (SSK6 did not express capacity to synthesise this toxic metabolite.
Full Text Available Fusarium species are wide-spread and known to be pathogenic agents to cultivated plants in various agroclimatic areas. During a four year investigation 10 Fusarium species and Microdochium nivale were isolated from wheat, barley, maize and soybean as well as from 10 weeds collected from 10 locations in Slavonia and Baranya. Fusarium graminearum was dominant on wheat and barley, F. moniliforme on maize and F. oxysporum on soybean. Regarding weeds, the presence of the following Fusarium species was established: F. graminearum on Amaranthus hybridus, Capsella bursa-pastoris, Lamium purpureum, Sorghum halepense and Urtica dioica, F. moniliforme on Abutilon theophrasti, F. subglutinans on Polygonum aviculare, F. avenaceum on Capsella bursa-pastoris, Rumex crispus and Matricaria sp., F. culmorum on Abutilon theophrasti, F. sporotrichioides on Polygonum aviculare, F. proliferatum and F. poae on Artemisia vulgaris. Pathogenicity test to wheat seedlings was done in our laboratory on winter wheat cultivars Slavonija and Demetra (totally 146 isolates. The most pathogenic species to wheat seedilings were F. graminearum, F. culmorum and F. sporotrichioides and the least pathogenic F. moniliforme, F. solani, F. oxysporum and F. poae. Pathogenicity test for wheat ears was done on genotypes Osk.8c9/3-94 and Osk.6.11/2 (totally 25 isolates. The results obtained by our investigation showed that there were no significant differences in pathogenicity of Fusarium species isolated from both cultivated plants and weeds. Weeds represent a constant source of inoculum of F. species for cultivated plants and they serve as epidemiologic bridges among vegetations.
Yarbrough, Melanie L; Lainhart, William; Burnham, Carey-Ann D
Nocardia species are the most commonly isolated aerobic actinomycetes from human clinical specimens. Our objective was to assess the identification of clinically relevant actinomycetes using the Bruker Biotyper MALDI-TOF system, including comparison of extraction methods, Biotyper library versions, score cutoffs, and media. Banked Streptomyces (n=10), Tsukamurella (n=2), and Nocardia isolates (n=60) were cultured and extracted using three methods: mycobacterial extraction, ethanol formic acid extraction, or direct on-target extraction. Following MALDI-TOF analysis, spectra were analyzed using versions 5 and 6 of the BDAL Biotyper library. Optimal species-level identifications for Nocardia were achieved using BDAL v6 at a score cutoff of ≥1.8 after direct extraction (49/60, 82%). Overall, the Biotyper platform with BDAL v6 accurately identified 12/16 species of Nocardia, demonstrating the utility of MALDI-TOF for identification of clinically relevant actinomycetes without the need for supplementation of the database. Copyright © 2017 Elsevier Inc. All rights reserved.
Latifa A. Al_husnan
Full Text Available Five strains of Streptomyces, namely S, N, W, E and C (designations should be mentioned in detail here isolated from the rhizosphere soil cultivated with palm Alajua (date, pressed dates, AlMedina city, Saudi Arabia, were induced to produce antibiotics. Antimicrobial activities were determined on solid medium supplemented with starch. The detection was based on the formation of transparent zones around colonies. The results indicated that isolates had antibacterial activities against Staphylococcus aureus, Bacillus cereus, B. subtilis, Pseudomonas aeruginosa and also showed antifungal activity against Candida albicans and Aspergillus niger. DNA extracted from five isolates was used as template for 16s rDNA gene amplification. The expected PCR size was 1.5 kbp;1.6 kbp; 1.25 kbp; 1.25kbp and 1.0 k bp for S, N, W, E and C isolates respectively using universal 16s rDNA gene primers using direct PCR. The isolates varied morphologically on the basis of spore color, aerial and substrate mycelium formation, and production of diffusible pigment. Isolates were tested under a microscope by using slide culture technique. The results indicate that the soil of this region is source of Streptomyces having antibacterial and antifungal activity and thus better utilization of these microorganisms as biological control agents.
Batta, Neha; Subudhi, Sanjukta; Lal, Banwari; Devi, Arundhuti
Lead is one of the four heavy metals that has a profound damaging effects on human health. In the recent past there has been an increasing global concern for development of sustainable bioremediation technologies for detoxification of lead contaminant. Present investigation highlights for lead biosorption by a newly isolated novel bacterial species; Achromobacter sp. TL-3 strain, isolated from activated sludge samples contaminated with heavy metals (collected from oil refinery, Assam, North-East India). For isolation of lead tolerant bacteria, sludge samples were enriched into Luria Broth medium supplemented separately with a range of lead nitrate; 250, 500, 750, 1000, 1250 and 1500 ppm respectively. The bacterial consortium that could tolerate 1500 ppm of lead nitrate was selected further for purification of lead tolerant bacterial isolates. Purified lead tolerant bacterial isolates were then eventually inoculated into production medium supplemented with ethanol and glycerol as carbon and energy source to investigate for bioflocculant production. Bioflocculant production was estimated by monitoring the potential of lead tolerant bacterial isolate to flocculate Kaolin clay in presence of 1% CaCl2. Compared to other isolates, TL-3 isolate demonstrated for maximum bioflocculant activity of 95% and thus was identified based on 16S rRNA gene sequence analysis. TL3 isolate revealed maximum homology (98%) with Achromobacter sp. and thus designated as Achromobacter sp. TL-3. Bioflocculant activity of TL-3 isolate was correlated with the change in pH and growth. Achromobacter sp. TL-3 has significant potential for lead biosorption and can be effectively employed for detoxification of lead contaminated waste effluents/waste waters.
Ergon, M C; Gülay, Z
Candida spp. has been the leading microorganism isolated from the urine specimens of patients hospitalized at the Anesthesiology and Reanimation intensive care unit (ICU) of Dokuz Eylul University Hospital, Izmir, since 1998. This study was undertaken to investigate the clonal relationship of Candida urine isolates in order to find the mode of spread among the patients. Epidemiological surveillance of 38 Candida albicans, 15 Candida tropicalis and 12 Candida glabrata recovered from the urine specimens of patients who were hospitalized in the ICU between June 11, 2000 and October 15, 2001 was carried out by antifungal susceptibility testing and randomly amplified polymorphic DNA (RAPD) analysis. Two short primers [Cnd3 (5'-CCAGATGCAC-3') and Cnd4 (5'-ACGGTACACT-3')] were used for RAPD. None of the isolates had high minimal inhibitory concentration (MIC) values (>1 microg ml(-1)) against amphotericin B with MIC50 values of 0.5 microg ml(-1), 0.5 microg ml(-1) and 0.125 microg ml(-1) for C. albicans, C. tropicalis and C. glabrata isolates, respectively. However, three C. glabrata isolates were resistant and one C. albicans and five C. glabrata isolates were dose-dependent susceptible (D-DS) to fluconazole. Among C. albicans isolates 19 and 20 patterns were detected with primers Cnd3 and Cnd4, respectively. When primers Cnd3 and Cnd4 were evaluated together, three and four genotypes were identified for C. tropicalis and C. glabrata isolates, respectively. Our results suggest that the source of C. albicans isolates was mostly endogenous. It is difficult to interpret the mode of spread of C. tropicalis and C. glabrata urine isolates as we obtained insufficient banding patterns for these species.
A relaxed (rel) mutant was found among 70 thiopeptin-resistant isolates of Streptomyces coelicolor A3(2) which arose spontaneously. The ability of the rel mutant to accumulate ppGpp during Casamino acid deprivation was reduced 10-fold compared to the wild-type. Analysis of the ribosomal proteins by two-dimensional PAGE revealed that the mutant lacked a ribosomal protein, tentatively designated ST-L11. It was therefore classified as a relC mutant. The mutant was defective in producing A-factor and the pigmented antibiotic prodigiosin, in both liquid and agar cultures, but produced agarase normally. Production of actinorhodin, another pigmented antibiotic, was also abnormal; it appeared suddenly in agar cultures after 10 d incubation. Although aerial mycelium still formed, its appearance was markedly delayed. Whereas liquid cultures of the parent strain accumulated ppGpp, agar cultures accumulated only trace amounts. Instead, a substance characterized only as an unidentified HPLC peak accumulated intracellularly in the late growth phase, just before aerial mycelium formation and antibiotic production. This substance did not accumulate in mutant cells. It was found in S. lividans 66 and S. parvulus, but not in seven other Streptomyces species tested. The significance of these observations, and the relationship of the mutant to earlier rel isolates of Streptomyces is discussed.
Keith F. Chater
Full Text Available About 2,500 papers dated 2014–2016 were recovered by searching the PubMed database for Streptomyces, which are the richest known source of antibiotics. This review integrates around 100 of these papers in sections dealing with evolution, ecology, pathogenicity, growth and development, stress responses and secondary metabolism, gene expression, and technical advances. Genomic approaches have greatly accelerated progress. For example, it has been definitively shown that interspecies recombination of conserved genes has occurred during evolution, in addition to exchanges of some of the tens of thousands of non-conserved accessory genes. The closeness of the association of Streptomyces with plants, fungi, and insects has become clear and is reflected in the importance of regulators of cellulose and chitin utilisation in overall Streptomyces biology. Interestingly, endogenous cellulose-like glycans are also proving important in hyphal growth and in the clumping that affects industrial fermentations. Nucleotide secondary messengers, including cyclic di-GMP, have been shown to provide key input into developmental processes such as germination and reproductive growth, while late morphological changes during sporulation involve control by phosphorylation. The discovery that nitric oxide is produced endogenously puts a new face on speculative models in which regulatory Wbl proteins (peculiar to actinobacteria respond to nitric oxide produced in stressful physiological transitions. Some dramatic insights have come from a new model system for Streptomyces developmental biology, Streptomyces venezuelae, including molecular evidence of very close interplay in each of two pairs of regulatory proteins. An extra dimension has been added to the many complexities of the regulation of secondary metabolism by findings of regulatory crosstalk within and between pathways, and even between species, mediated by end products. Among many outcomes from the application of
Sato, Shingo; Kabeya, Hidenori; Miura, Tatsuya; Suzuki, Kazuo; Bai, Ying; Kosoy, Michael; Sentsui, Hiroshi; Kariwa, Hiroaki; Maruyama, Soichi
The prevalence of Bartonella species was investigated among wild carnivores of the suborder Caniformia, including 15 Japanese badgers (Meles anakuma), 8 Japanese martens (Martes melampus), 2 Japanese weasels (Mustela itatsi), 1 Siberian weasel (Mustela sibirica), 171 raccoon dogs (Nyctereutes procyonoides), and 977 raccoons (Procyon lotor) in Japan. Bartonella bacteria were isolated from one Japanese badger (6.7%) and from one Japanese marten (12.5%); however, no Bartonella species was found in other representatives of Caniformia. Phylogenetic analysis was based on concatenated sequences of six housekeeping genes (16S rRNA, ftsZ, gltA, groEL, ribC, and rpoB) and sequence of the 16S-23S internal transcribed spacer region. The sequence analysis indicated that the isolate derived from the Japanese badger (strain JB-15) can represent a novel Bartonella species and the isolate from the Japanese marten (strain JM-1) was closely related to Bartonella washoensis. This is the first report on isolation of Bartonella from badger and marten.
Guerra, Anderson; Filpponen, Ilari; Lucia, Lucian A; Argyropoulos, Dimitris S
Milled wood lignin (MWL), cellulolytic enzyme lignin (CEL), and enzymatic mild acidolysis lignin (EMAL) were isolated from different wood species and characterized by various techniques. The EMAL protocol offered gravimetric lignin yields 2-5 times greater than those of the corresponding MWL and CEL. The purities of the EMALs were 3.75-10.6% higher than those of their corresponding CELs, depending upon the wood species from which they were isolated. Molecular weight analyses showed that the EMAL protocol isolates lignin fractions that are not accessed by the other procedures evaluated, while 31P NMR spectroscopy revealed that MWL is more condensed and bears more phenolic hydroxyl groups than EMAL and CEL. The yields and purities of EMAL, MWL, and CEL from hardwood were greater than those obtained for the examined softwoods. Structural details obtained by DFRC (derivatization followed by reductive cleavage)/31P NMR revealed different contents of condensed and uncondensed beta-O-aryl ether structures, dibenzodioxocins, and condensed and uncondensed phenolic hydroxyl and carboxylic acid groups within lignins isolated from different wood species.
Soares, José C; Marques, M Rosário; Tavaria, Freni K; Pereira, Joana O; Malcata, F Xavier; Pintado, Manuela M
The level and the diversity of the staphylococcal community occurring in the environment and dairy products of a small manufacturing dairy plant were investigated. Species identification was performed using different molecular methods, viz. Multiplex-PCR, amplified ribosomal DNA restriction analysis (ARDRA), and sodA gene sequencing. The main species encountered corresponded to Staphylococcus equorum (41 isolates, 39.0%), S. saprophyticus (28 isolates, 26.7%) and S. epidermidis (15 isolates, 14.3%). Additionally, low incidence of enterotoxin genes was obtained, with only 9 strains (8.6%) being positive for one or more toxin genes. With regard to antimicrobial resistance, 57.1% of the isolates showed at least resistance against one antibiotic, and 28.6% were multi-resistant, which might accomplish resistance for up to 6 antibiotics simultaneously. These results provided evidence that the presence of Staphylococcus species in dairy environment are mostly represented by S. equorum and S. saprophyticus, and illustrate that carrying antimicrobial resistance genes has become reasonably widespread in cheese and dairy environment.
Full Text Available Introduction. Campylobacter is one of the leading bacterial causes of food-borne disease. The prevalence of Campylobacter species resistant to antimicrobial agents is increasing. This study is intended to determine prevalence and antimicrobial susceptibility patterns of Campylobacter species among under-five children with diarrhea. Methodology. A cross-sectional study was conducted among 227 under-five children with diarrhea from July to October 2012 at Jimma town. Isolation and identification of Campylobacter species were performed using standard bacteriological techniques. Antimicrobial susceptibility test was performed following standard protocol. Chi-square and Fisher’s exact tests were used for analysis. Results. From 227 under-five children, 16.7% were positive for Campylobacter spp.; isolates, C. jejuni, C. coli, and C. lari, accounted for 71.1%, 21.1%, and 7.9%, respectively. Higher rate of resistance was observed to ampicillin 76.3%, trimethoprim-sulfamethoxazole (68.4%, tetracycline (39.5%, chloramphenicol (31.6%, clindamycin (26.3%, and doxycycline (23.7%. Erythromycin, ciprofloxacin, gentamicin, norfloxacin, and nalidixic acid were effective for more than 80% of the isolates. Multiple drug resistance was observed among 78.9% of all the three spp. Conclusions. Isolation rate of Campylobacter spp. was high. C. lari was reported for the first time at this study area. Higher rate of resistance was observed to the commonly used drugs.
Feng-Juan Wang; Dai Zhang; Zhao-Hui Liu; Wen-Xiang Wu; Hui-Hui Bai; Han-Yu Dong
Background:Vulvovaginal candidiasis (VVC) was a common infection associated with lifelong harassment of woman's social and sexual life.The purpose of this study was to describe the species distribution and in vitro antifungal susceptibility of Candida species (Candida spp.) isolated from patients with WC over 8 years.Methods:Species which isolated from patients with WC in Peking University First Hospital were identified using chromogenic culture media.Susceptibility to common antifungal agents was determined using agar diffusion method based on CLSI M44-A2 document.SPSS software (version 14.0,Inc.,Chicago,IL,USA) was used for statistical analysis,involving statistical description and Chi-square test.Results:The most common strains were Candida (C.) albicans,80.5％ (n =1775) followed by C.glabrata,18.1％ (n =400).Nystatin exhibited excellent activity against all species (＜4％ resistant [R]).Resistance to azole drugs varied among different species.C albicans:clotrimazole (3.1％ R) ＜ fluconazole (16.6％ R) ＜ itraconazole (51.5％ R) ＜ miconazole (54.0％ R);C.glabrata:miconazole (25.6％ R) ＜ clotrimazole (50.5％ R) ＜ itraconazole (61.9％ R) ＜ fluconazole (73.3％ R);Candida krusei:clotrimazole (0 R) ＜ fluconazole (57.7％ R) ＜ miconazole (73.1％ R) ＜ itraconazole (83.3％ R).The susceptibility offluconazole was noticeably decreasing among all species in the study period.Conclusions:Nystatin was the optimal choice for the treatment of WC at present.The species distribution and in vitro antifungal susceptibility of Candida spp.isolated from patients with VVC had changed over time.
Wang, Feng-Juan; Zhang, Dai; Liu, Zhao-Hui; Wu, Wen-Xiang; Bai, Hui-Hui; Dong, Han-Yu
Background: Vulvovaginal candidiasis (VVC) was a common infection associated with lifelong harassment of woman's social and sexual life. The purpose of this study was to describe the species distribution and in vitro antifungal susceptibility of Candida species (Candida spp.) isolated from patients with VVC over 8 years. Methods: Species which isolated from patients with VVC in Peking University First Hospital were identified using chromogenic culture media. Susceptibility to common antifungal agents was determined using agar diffusion method based on CLSI M44-A2 document. SPSS software (version 14.0, Inc., Chicago, IL, USA) was used for statistical analysis, involving statistical description and Chi-square test. Results: The most common strains were Candida (C.) albicans, 80.5% (n = 1775) followed by C. glabrata, 18.1% (n = 400). Nystatin exhibited excellent activity against all species (<4% resistant [R]). Resistance to azole drugs varied among different species. C. albicans: clotrimazole (3.1% R) < fluconazole (16.6% R) < itraconazole (51.5% R) < miconazole (54.0% R); C. glabrata: miconazole (25.6% R) < clotrimazole (50.5% R) < itraconazole (61.9% R) < fluconazole (73.3% R); Candida krusei: clotrimazole (0 R) < fluconazole (57.7% R) < miconazole (73.1% R) < itraconazole (83.3% R). The susceptibility of fluconazole was noticeably decreasing among all species in the study period. Conclusions: Nystatin was the optimal choice for the treatment of VVC at present. The species distribution and in vitro antifungal susceptibility of Candida spp. isolated from patients with VVC had changed over time. PMID:27174323
Nazemi, A. (PhD
Full Text Available Background and Objective: Recognizing and using of isolated phytase in the soil microorganisms are paramount importance to produce the Phytase enzyme utilized commercially in different industries. This study was conducted to recognize different bacillus species which are Phytase producers and detection of the gene that can produce this enzyme. Material and Methods: Soil samples were gathered through different parts of mountainous areas. The early isolation of bacillus was carried out in Bacillus Medium Agar. After isolating the bacteria and genome extraction, the responsible gene of enzyme producer recognized and amplified by PCR method. The size of this protein and the optimal production situation in supplemental exploitation such as SDS-PAGE and the enzymatic activity of its size were evaluated. Results: Of 40 samples, one bacterium secreting Phytase enzyme was isolated. This bacterium was sequenced and recognized Bacillus Sobtlis species that is classified in STR Genus. The size of protein phytase produced by this gene was about 45 KD and the enzyme activity at 55 degrees was measured about 5.65 in wavelength of 415 NM. The phytase gene with the size of 1200 bp was propagated. Conclusion: the microorganisms, in natural conditions, produce Phytase enzyme in limited amount and with the quality appropriate to microorganisms. Thus, isolating the bacilli producing Phytase enzyme and purifying this protein are highly significant. Key words: Bacillus Subtilis; Phytase; SDS-PAGE; Enzymatic Activity; Polymerization Chain Reaction
Cai, Y; Kumai, S; Ogawa, M; Benno, Y; Nakase, T
Pediococcus species isolated from forage crops were characterized, and their application to silage preparation was studied. Most isolates were distributed on forage crops at low frequency. These isolates could be divided into three (A, B, and C) groups by their sugar fermentation patterns. Strains LA 3, LA 35, and LS 5 are representative isolates from groups A, B, and C, respectively. Strains LA 3 and LA 35 had intragroup DNA homology values above 93.6%, showing that they belong to the species Pediococcus acidilactici. Strain LS 5 belonged to Pediococcus pentosaceus on the basis of DNA-DNA relatedness. All three of these strains and strain SL 1 (Lactobacillus casei, isolated from a commercial inoculant) were used as additives to alfalfa and Italian ryegrass silage preparation at two temperatures (25 and 48 degrees C). When stored at 25 degrees C, all of the inoculated silages were well preserved and exhibited significantly (P ammonia-nitrogen content, gas production, and dry matter loss and significantly (P < 0.05) higher lactate content than the control, but silages inoculated with LS 5 and SL 1 were of poor quality. P. acidilactici LA 3 and LA 35 are considered suitable as potential silage inoculants.
Olumide A. Odeyemi
Full Text Available This study aimed to investigate antibiotics resistance pattern and phenotyping of Aeromonas species isolated from different aquatic sources in Melaka, Malaysia. A total of 53 Aeromonas species were isolated from the following sources: sediment (n = 13, bivalve (n = 10, sea cucumber (n = 16 and sea water (n = 14 and resistance to 12 antibiotics – Tetracycline (30 μg, Kanamycin (30 μg, Oxytetracycline (30 μg, Ampicillin (10 μg, Streptomycin (10 μg, Gentamicin (10 μg, Sulphamethoxazole (25 μg, Nalixidic acid (30 μg, Trimethoprim (1.25 μg, Novobiocin (5 μg, Penicilin (10 μg and Chloramphenicol (10 μg was tested. The results obtained from this study reveal multi drug resistance pattern among the isolates. All the isolates were completely resistant to Ampicillin, Novobiocin, Sulphamethoxazole and Trimethoprim, respectively but susceptible to Tetracycline (100%, Kanamycin (5.7%, Gentamicin (5.7% and Oxytetracycline (24.5%. Antibiotics phenotyping of the bacteria revealed 21 different phenotypes among the isolates.
Ziedins, A C; Chomel, B B; Kasten, R W; Kjemtrup, A M; Chang, C-C
Bartonella spp. are endemic in wild rodents in many parts of the world. A study conducted in two northern California counties (Sonoma and Yolo) sampling California ground squirrels (Otospermophilus beecheyi) and four other rodent species (Peromyscus maniculatus, P. boylii, P. truei and Neotoma fuscipes) led to the isolation of small Gram-negative bacilli which were identified as Bartonella spp. based on colony morphology, polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and partial gene sequencing. Overall, Bartonella spp. were isolated from the blood of 71% (32/45) of the ground squirrels and one third (22/66) of the other rodents. PCR-RFLP analysis of the gltA and 16S rRNA genes yielded seven unique profiles, four for the ground squirrels and three for the other rodents. Isolates from each PCR-RFLP profiles were submitted for partial sequencing. Ground squirrel isolates were most closely related to B. washoensis, whereas the other rodent isolates were closest to B. vinsonii subsp. vinsonii and B. vinsonii subsp. arupensis. Two of these three species or subspecies are known zoonotic agents.
Odeyemi, Olumide A; Ahmad, Asmat
This study aimed to investigate antibiotics resistance pattern and phenotyping of Aeromonas species isolated from different aquatic sources in Melaka, Malaysia. A total of 53 Aeromonas species were isolated from the following sources: sediment (n = 13), bivalve (n = 10), sea cucumber (n = 16) and sea water (n = 14) and resistance to 12 antibiotics - Tetracycline (30 μg), Kanamycin (30 μg), Oxytetracycline (30 μg), Ampicillin (10 μg), Streptomycin (10 μg), Gentamicin (10 μg), Sulphamethoxazole (25 μg), Nalixidic acid (30 μg), Trimethoprim (1.25 μg), Novobiocin (5 μg), Penicilin (10 μg) and Chloramphenicol (10 μg) was tested. The results obtained from this study reveal multi drug resistance pattern among the isolates. All the isolates were completely resistant to Ampicillin, Novobiocin, Sulphamethoxazole and Trimethoprim, respectively but susceptible to Tetracycline (100%), Kanamycin (5.7%), Gentamicin (5.7%) and Oxytetracycline (24.5%). Antibiotics phenotyping of the bacteria revealed 21 different phenotypes among the isolates.
Sullivan, J T; Eardly, B D; van Berkum, P; Ronson, C W
Previously, we found that genetically diverse rhizobia nodulating Lotus corniculatus at a field site devoid of naturalized rhizobia had symbiotic DNA regions identical to those of ICMP3153, the inoculant strain used at the site (J. T. Sullivan, H. N. Patrick, W. L. Lowther, D. B. Scott, and C. W. Ronson, Proc. Natl. Acad. Sci. USA 92:8985-8989, 1995). In this study, we characterized seven nonsymbiotic rhizobial isolates from the rhizosphere of L. corniculatus. These included two from plants at the field site sampled by Sullivan et al. and five from plants at a new field plot adjacent to that site. The isolates did not nodulate Lotus species or hybridize to symbiotic gene probes but did hybridize to genomic DNA probes from Rhizobium loti. Their genetic relationships with symbiotic isolates obtained from the same sites, with inoculant strain ICMP3153, and with R. loti NZP2213T were determined by three methods. Genetic distance estimates based on genomic DNA-DNA hybridization and multilocus enzyme electrophoresis were correlated but were not consistently reflected by 16S rRNA nucleotide sequence divergence. The nonsymbiotic isolates represented four genomic species that were related to R. loti; the diverse symbiotic isolates from the site belonged to one of these species. The inoculant strain ICMP3153 belonged to a fifth genomic species that was more closely related to Rhizobium huakuii. These results support the proposal that nonsymbiotic rhizobia persist in soils in the absence of legumes and acquire symbiotic genes from inoculant strains upon introduction of host legumes.
Khor, Wei Ching; Puah, Suat Moi; Tan, Jin Ai Mary Anne; Puthucheary, S D; Chua, Kek Heng
Gram-negative bacilli of the genus Aeromonas are primarily inhabitants of the aquatic environment. Humans acquire this organism from a wide range of food and water sources as well as during aquatic recreational activities. In the present study, the diversity and distribution of Aeromonas species from freshwater lakes in Malaysia was investigated using glycerophospholipid-cholesterol acyltransferase (GCAT) and RNA polymerase sigma-factor (rpoD) genes for speciation. A total of 122 possible Aeromonas strains were isolated and confirmed to genus level using the API20E system. The clonality of the isolates was investigated using ERIC-PCR and 20 duplicate isolates were excluded from the study. The specific GCAT-PCR identified all isolates as belonging to the genus Aeromonas, in agreement with the biochemical identification. A phylogenetic tree was constructed using the rpoD gene sequence and all 102 isolates were identified as: A. veronii 43%, A. jandaei 37%, A. hydrophila 6%, A. caviae 4%, A. salmonicida 2%, A. media 2%, A. allosaccharophila 1%, A. dhakensis 1% and Aeromonas spp. 4%. Twelve virulence genes were present in the following proportions--exu 96%, ser 93%, aer 87%, fla 83%, enolase 70%, ela 62%, act 54%, aexT 33%, lip 16%, dam 16%, alt 8% and ast 4%, and at least 2 of these genes were present in all 102 strains. The ascV, aexU and hlyA genes were not detected among the isolates. A. hydrophila was the main species containing virulence genes alt and ast either present alone or in combination. It is possible that different mechanisms may be used by each genospecies to demonstrate virulence. In summary, with the use of GCAT and rpoD genes, unambiguous identification of Aeromonas species is possible and provides valuable data on the phylogenetic diversity of the organism.
Wei Ching Khor
Full Text Available Gram-negative bacilli of the genus Aeromonas are primarily inhabitants of the aquatic environment. Humans acquire this organism from a wide range of food and water sources as well as during aquatic recreational activities. In the present study, the diversity and distribution of Aeromonas species from freshwater lakes in Malaysia was investigated using glycerophospholipid-cholesterol acyltransferase (GCAT and RNA polymerase sigma-factor (rpoD genes for speciation. A total of 122 possible Aeromonas strains were isolated and confirmed to genus level using the API20E system. The clonality of the isolates was investigated using ERIC-PCR and 20 duplicate isolates were excluded from the study. The specific GCAT-PCR identified all isolates as belonging to the genus Aeromonas, in agreement with the biochemical identification. A phylogenetic tree was constructed using the rpoD gene sequence and all 102 isolates were identified as: A. veronii 43%, A. jandaei 37%, A. hydrophila 6%, A. caviae 4%, A. salmonicida 2%, A. media 2%, A. allosaccharophila 1%, A. dhakensis 1% and Aeromonas spp. 4%. Twelve virulence genes were present in the following proportions--exu 96%, ser 93%, aer 87%, fla 83%, enolase 70%, ela 62%, act 54%, aexT 33%, lip 16%, dam 16%, alt 8% and ast 4%, and at least 2 of these genes were present in all 102 strains. The ascV, aexU and hlyA genes were not detected among the isolates. A. hydrophila was the main species containing virulence genes alt and ast either present alone or in combination. It is possible that different mechanisms may be used by each genospecies to demonstrate virulence. In summary, with the use of GCAT and rpoD genes, unambiguous identification of Aeromonas species is possible and provides valuable data on the phylogenetic diversity of the organism.
Ahmed M; Singh MN; Bera AK; Bandyopadhyay S; Bhattacharya D
Gastrointestinal(GI)parasitism is the most serious constraint throughout the world in small ruminants which causes significant production loss in animals.GI parasites are major contributor to reduce productivity in terms of meat, milk and wool in animals. Control ofGI parasite is done primarily by anthelmintic treatment where choice and schedule of treatment is done after identification and quantitation of individual parasite. Identification ofGI parasites is done through microscopic method by identifying specific morphological characteristics of egg and larva (L3). Since most of parasite eggs are having similar morphological characteristics, identification up to species level through microscopy is not possible in most of cases. To address this issue, molecular techniques are the viable alternative for identification of species as well as molecular level differences within a species (isolates) of parasites. DifferentDNA based molecular techniquesviz.PCR, AFLP, RAPD, RFLP, PCR-SSCP, real timePCR, DNAmicroarrayetc. have been used for identification and to assess the genetic diversity among parasite population. For identification of species, the characteristic sequence of genomicDNAof different species should differ to allow the delineation of species, but at the same time, no/minor variation within the species should exist. In contrast, for purpose of identifying population variants (strains/isolates), a considerable degree of variation in the sequence should exist within a species. Various target regions, including nuclear ribosomal DNA (rDNA), mitochondrial DNA(mtDNA) or repetitiveDNA elements (microsatellite loci), which show considerable variation in the number of repeats within individuals have been employed to achieve the identification of parasites species or strain.
Fernández-Mazuecos, M; Vargas, P
The lack of a comparative approach makes it impossible to determine the main factors influencing colonization and evolution in plants. Here we conducted the first comparative study of a characteristic Mediterranean lineage (white-flowered Cistus) taking advantage of its well-known phylogenetic relationships. A two-scale approach was applied to address the hypothesis of higher levels of isolation in mountain than in lowland species. First, a time-calibrated phylogeny using plastid sequences of Cistaceae suggested that the origin of Cistus species postdated both the refilling of the Mediterranean Sea (5.59-5.33 Ma) and the onset of the Mediterranean climate (3.2 Ma). Two hundred and sixty-three additional, plastid sequences from 111 populations showed different numbers of haplotypes in C. laurifolius (7), C. monspeliensis (2) and C. salviifolius (7). Although haplotype sharing among disjunct populations was observed in all species, phylogeographic analyses revealed haplotype lineages exclusive to Europe or Africa only in the mountain species (C. laurifolius). Isolation by either geographical distance or sea barriers was not significantly supported for the lowland species (C. monspeliensis; C. ladanifer from a previous study). The same is true for the less habitat-specific species of the lineage (C. salviifolius). Comparative phylogeography of the Cistus species leads us to interpret a general pattern of active colonization surpassing Mediterranean barriers. In contrast, ecological conditions (precipitation, temperature, soils) appear to have determined the distribution of the Cistus species of Mediterranean mountains. This study further provides molecular evidence for multiple colonization patterns in the course of successful adaptation of Cistus species to Mediterranean habitats.
Levican, Arturo; Collado, Luis; Aguilar, Carmen; Yustes, Clara; Diéguez, Ana L; Romalde, Jesús L; Figueras, Maria José
A group of ten Arcobacter isolates (Gram negative, slightly curved motile rods, oxidase positive) was recovered from mussels (nine) and from clams (one). These isolates could not be assigned to any known species using the molecular identification methods specific for this genus (16S rDNA-RFLP and m-PCR). The aim of this study is to establish the taxonomic position of these isolates. The 16S rRNA gene sequence similarity of mussel strain F4(T) to the type strains of all other Arcobacter species ranged from 91.1% to 94.8%. The species most similar to the clams' strain F67-11(T) were Arcobacter defluvii (CECT 7697(T), 97.1%) and Arcobacter ellisii (CECT 7837(T), 97.0%). On the basis of phylogenetic analyses with 16S rRNA, rpoB, gyrB and hsp60 genes, the mussel and clam strains formed two different, new lineages within the genus Arcobacter. These data, together with their different phenotypic characteristics and MALDI-TOF mass spectra, revealed that these strains represent two new species, for which the names Arcobacter bivalviorum (type strain F4(T)=CECT 7835(T)=LMG 26154(T)) and Arcobacter venerupis (type strain F67-11(T)=CECT 7836(T)=LMG 26156(T)) are proposed.
Dahal, Bibha; NandaKafle, Gitanjali; Perkins, Lora; Brözel, Volker S
Biological Nitrogen Fixation is critical for ecosystem productivity. Select members of Bacteria and Archaea express a nitrogenase enzyme complex that reduces atmospheric nitrogen to ammonia. Several nitrogen fixing bacteria form symbiotic associations with plants, but free-living diazotrophs also contribute a substantial amount of nitrogen to ecosystems. The aim of this study was to isolate and characterize free-living diazotrophs in arid lands of South Dakota Badlands. Samples were obtained from sod tables and the surrounding base in spring and fall. Diazotrophs were isolated on solid nitrogen free medium (NFM) under hypoxic conditions, and their16S rRNA and nifH genes sequenced. nifH was also amplified directly from soil DNA extracts. The 16S rRNA gene data indicated a diversity of putative free-living diazotrophs across 4 phyla (Actinomycetes, Proteobacteria, Bacteroidetes, and Firmicutes), but ∼50% of these clustered with Streptomyces. These Streptomyces isolates grew in liquid NFM in an ammonia-depleted environment. Only 5 of these yielded a nifH gene product using the PolF/PolR primer set. Four of these aligned with nifH of the cyanobacteria Scytonema and Nostoc, and the other one aligned with nifH of Bradyrhizobium. Six selected Streptomyces isolates, three of which were nifH positive by PCR, all indicated (15)N2 incorporation, providing strong support of nitrogen fixation. All nifH amplicons from soil DNA extract resembled Cyanobacteria. This is the first known report of diazotrophic Streptomyces, other than the thermophilic, autotrophic S. thermoautotrophicus. nifH genes of these Streptomyces were related to those from Cyanobacteria. It is possible that the cyanobacteria-like nifH amplicons obtained from soil DNA were associated with Streptomyces.
Puławska, Joanna; Willems, Anne; De Meyer, Sofie E; Süle, Sandor
Five Gram-negative, rod-shaped, non-spore-forming bacteria were isolated from galls on different plant species in Hungary: strain 39/7(T) from Prunus cerasifera Myrobalan, s