A Chitinolytic Actinomycete Complex in Chernozem Soil

A chitinolytic actinomycete complex in chernozem soil has a specific taxonomic composition, which differs from that of the actinomycete complex typically isolated on standard nutrient media containing sugars and organic acids as carbon sources. The actinomycete complex that was isolated by using nutrient media with chitin as the source of carbon and nitrogen was dominated by representatives of the genus Streptosporangium, and the actinomycete complex that was isolated by using nutrient media with sugars and organic acids as the carbon sources was dominated by representatives of the genus Streptomyces. The confirmation of the ability of actinomycetes to utilize chitin as a sole source of carbon and nitrogen came from the augmented length and biomass of the mycelium, the increased number and biomass of the actinomycete spores, the production of carbon dioxide, and the accumulation of NH₄ ⁺ ions in the culture liquid of the actinomycetes grown in the nutrient media with chitin.     

Muddy odour: a problem associated with extreme eutrophication

The results of a 4-year study in a hypereutrophic bay, Kaupunginselkä Bay at Porvoo on the south coast of Finland, indicated a significant correlation between muddy odour in bream (Abramis brama) and the amount of the blue-green alga Oscillatoria agardhii in the phytoplankton. This algal strain has previously been shown to produce the muddy-smelling compound geosmin. The numbers of muddy-smelling actinomycetes in the water and sediments of the study area were not clearly related to muddy odour in fish, nor to phytoplankton biomass. In the hypereutrophic L. Tuusulanjärvi, muddy odour in bream and pikeperch (Stizostedion lucioperca) was also related to the amounts of blue-green algae in the phytoplankton.     

Bioactive compounds from marine actinomycetes

Actinomycetes are one of the most efficient groups of secondary metabolite producers and are very important from an industrial point of view. Among its various genera, Streptomyces, Saccharopolyspora, Amycolatopsis, Micromonospora and Actinoplanes are the major producers of commercially important biomolecules. Several species have been isolated and screened from the soil in the past decades. Consequently the chance of isolating a novel actinomycete strain from a terrestrial habitat, which would produce new biologically active metabolites, has reduced. The most relevant reason for discovering novel secondary metabolites is to circumvent the problem of resistant pathogens, which are no longer susceptible to the currently used drugs. Existence of actinomycetes has been reported in the hitherto untapped marine ecosystem. Marine actinomycetes are efficient producers of new secondary metabolites that show a range of biological activities including antibacterial, antifungal, anticancer, insecticidal and enzyme inhibition. Bioactive compounds from marine actinomycetes possess distinct chemical structures that may form the basis for synthesis of new drugs that could be used to combat resistant pathogens.     

Peculiarities of Growth and Morphological Differentiation of Acidophilic and Neutrophilic Soil Streptomycetes

Acidophilic actinomycetes are shown to possess a special mechanism of adaptation to low pH conditions, which shows up in their ability to grow faster on acidified than on neutral media and to adjust the pH of the medium to a level favorable for the formation of aerial mycelium and spores. On nutrient media with pH below 5, neutrophilic actinomycetes either fail to grow or grow much slower than on neutral media; they do not alkalize the medium and do not form aerial mycelium.     

Evidence of a rheotactic component in the odour search behaviour of freshwater eels

The detection of food odour by the freshwater eels, Anguilla australis and Anguilla dieffenbachii released a behavioural response to flow that resulted in direct upstream movement toward the odour source. Changes in various orientation parameters were observed as eels neared the source. Eels had substantially lower swimming velocities and considerably more variable heading and course angles close to the odour source (≤0·9 m) than further away (>0·9 m). Observed changes in orientation parameters were primarily due to changes in the behaviour of a searcher following odour loss. Cross-stream movements were initiated when the eel moved beyond the lateral margins of the odour plume. The behavioural switch from odour-conditioned rheotaxis to cross-stream casting following odour loss occurred more frequently close to the odour source where the plume was most narrow. Odour-conditioned rheotaxis enables the searcher to move quickly and efficiently toward the odour source without the need to extract directional information from a highly intermittent and complex chemical signal.     

Development of an intergeneric conjugal transfer system for rimocidin‐producing Streptomyces rimosus

Aims: To develop an intergeneric conjugation system for rimocidin‐producing Streptomyces rimosus. Methods and Results: High efficiencies of conjugation [10^?2–10^?3 transconjugants/recipient colony forming units (CFU)] were obtained when spores of S. rimosus were heat treated at 40°C for 10 min prior to mixing with E. coli ET12567(pUZ8002/pIJ8600) as donor. Mycelium from liquid grown cultures of S. rimosus could also be used as recipient instead of spores, with 24‐h cultures giving optimal results. TSA (Oxoid) medium containing 10 m mol l^?1 MgCl₂ was the preferred medium for conjugation. Southern hybridization was used to confirm that transconjugants of S. rimosus contained a single copy of pIJ8600 integrated at a unique chromosomal attachment site (attB). The transconjugants exhibited a high stability of plasmid integration and showed strong expression of green fluorescent protein when using pIJ8655 as the conjugative vector. Conclusion: Intergeneric conjugation between E. coli and S. rimosus was achieved at high efficiency using both spores and mycelium. Significance and Impact of the Study: The conjugation system developed provides a convenient gene expression system for S. rimosus R7 and will enable the genetic manipulation of the rimocidin gene cluster.     

Isolation and characterization of alachlor-degrading actinomycetes from soil

Alachlor (2-cloro-N-(methoxymethyl)-N-(2,6-diethylphenyl)-acetamide) is an extremely toxic and highly mobile herbicide that is widely used for pre-emergence control of grasses and weeds in many commercial crops in Brazil. In order to select soil actinomycetes able to degrade this herbicide, fifty-three actinomycete strains were isolated from soil treated with alachlor using selective conditions and subjected to in vitro degradation assays. Sixteen isolates were shown to be tolerant to high concentrations of the herbicide (up to 720 mg L^-1), and six of these were able to grow and degrade 50 alachlor (72 mg L^-1) in mineral salts medium. Morphological and phylogenetic analysis enabled the assignment of the alachlor-degrading strains to the genus Streptomyces. Strain LS151 was related to the type strains of Streptomyces capoamus/Streptomyces galbus, whereas strains LS143 and LS153 were related to Streptomyces bikiniensis. The remaining strains, LS166, LS177 and LS182, were similar in morphological features and recovered in a single cluster based on 16S rDNA sequence analysis, but shown to be distinct on the basis of genomic fingerprint data (rep-PCR). Though a definitive taxonomic assignment of alachlor-degrading strains was not possible, these data indicate that ability to degrade this pesticide was detected in different Streptomyces taxa.     

灯台树内生放线菌

放线菌一直是用于生产抗生素最多的微生物类群,临床和农牧业上应用的抗生素有60%以上就是由放线菌生产的[1],但随着从来源于普通土壤环境的放线菌中筛选到新化合物几率的不断下降,人们已经逐步将注意力转向了一些生活在特殊生境的微生物[2]。由于植物内生菌在植物体内经过与植物宿主的长期协同进化,使植物内生菌能够产生结构新颖、功能     

Colonization of plant rhizosphere by actinomycetes of different genera

The survival of environmental isolates of actinomycetes introduced with the seeds of agricultural plants in root-free soil and in the rhizosphere and rhizoplane was studied. Different strategies of colonization of the rhizosphere were revealed for the representatives of the genera Streptomyces, Micromonospora, and Streptosporangium, organisms typical for the moderate climate rhizosphere. The plants of winter rye (Secale cereale L.) inoculated with actinomycetes were shown to have growth advantages, while the cow clover plants (Trifolium pratense L.) had no growth advantages compared to uninoculated plants. The role of the plant component in the interaction with mycelial prokaryotes is discussed.     

植物内生放线菌及其生理活性物质研究进展

植物内生放线菌是一类具有巨大开发潜力的新微生物资源。目前从许多活体植物组织内已分离到种类众多的植物内生放线菌,已有的研究表明植物内生放线菌能产生许多重要的生理活性物质,如抗生素类物质、植物生长促进剂、植物生长抑制剂以及具有新特性的酶类。植物内生放线菌在农业生产、医药新药的筛选上显示出广阔的应用前景。     

Intergeneric conjugal gene transfer from Escherichia coli to the sweet potato pathogen Streptomyces ipomoeae

Aims:  To develop an effective gene transfer system for Streptomyces ipomoeae , the causative agent of soil rot disease of sweet potatoes.
Methods and Results:  Of the several methods investigated, introduction of genes into S. ipomoeae could only be achieved using intergeneric conjugation from Escherichia  coli . However, even results for that method varied greatly and were dependent on using particular media for S. ipomoeae spore preparation and conjugation. Transconjugant to recipient ratios as great as 4·1 × 10⁻⁵ were achieved when International Streptomyces Project Medium 4 was used for both sporulation and conjugation protocols. Both site-specifically integrating and autonomously replicating plasmids could be introduced and maintained in S. ipomoeae, and plasmids could be introduced with approximate equivalent frequencies from either methyl-proficient or methyl- deficient E. coli donors; the latter result indicates a likely absence of relevant methyl-specific restriction in S. ipomoeae .
Conclusions:  Efficient transfer of genes into S. ipomoeae was achieved here by using an optimized intergeneric mating procedure.
Significance and Impact of the Study:  The described protocol will facilitate further genetic manipulation of this agriculturally important pathogen.     

土壤放线菌分离方法的初步研究

本文对土壤放线菌的一种新分离技术进行了研究。结果表明,用加有氟哌酸、青霉素、制霉菌素的培养基,可以选择性地从土壤中分离放线菌,０．０５％ＳＤＳ（十二烷基碘酸钠）在４０℃自理土壤样品,可以促进放线菌的孢子萌发而获得更多的放线菌株,进一步提高放线菌的分离效果。     

Biosystematics and diversity amongst novel carboxydotrophic actinomycetes

Fifty-four carboxydotrophic actinomycetes isolated from soils and composts were compared through 119 unit characters with representative mesophilic and thermophilic streptomycetes. The data were examined using the Jaccard, pattern and simple matching coefficients and clustering achieved using the unweighted pair group method with arithmetic averages algorithm. Acceptable cophenetic correlation and test error values allowed confidence to be placed in the resultant numerical taxonomies. The carboxydotrophic actinomycetes, which were distinct from cluster-groups corresponding to the mesophilic and thermophilic streptomycetes, formed two major cluster-groups the members of which were examined for the presence of diagnostic chemical markers. All but two of the carboxydotrophic actinomycetes had a profile of chemical properties consistent with their assignment to the genusStreptomyces. Quantitative fatty acid data were examined using the SIMCA package and the two statistically significant groups obtained corresponded with the cluster-groups circumscribed in the numerical phenetic analysis. Members of the two groups were also distinguished on the basis of their phospholipid composition. The two strains that containedmeso- as opposed to LL-diaminopimelic acid in their peptidoglycan also showed a distinct chemotaxonomic profile. It was concluded that the carboxydotrophic actinomycetes form a novel and taxonomically diverse group.     

Rapid detection of `rare' actinomycetes in environmental samples

Natural products continue to be a useful source of new leads for the pharmaceutical industry. Actinomycetes are prolific producers of natural products and one strategy to increase the possibility of discovering novel chemical entities is to screen actinomycetes considered 'rare' in the environment and previously under-represented in natural product screening collections. We describe a method using bacteriophage as a marker to detect these actinomycetes in environmental samples. This method allows samples to be pre-screened for the presence of target actinomycetes before lengthy isolation programmes are undertaken.     

嗜酸丝状放线菌的选择性分离与多样性

摘要:【目的】针对酸性土壤中的嗜酸丝状放线菌,建立有效的选择性分离方法,并了解其多样性。【方法】用不同的样品预处理方式和分离培养基,并添加不同的抑制剂进行分离;根据放线菌的菌落数和出菌率确定最佳分离方法组合。采用最佳分离方法对从江西采集的17份酸性土壤样品进行分离;根据培养特征对分离菌株进行分群,进一步通过对各类群的显微形态观察和pH梯度生长实验确定代表菌株;对代表菌株进行16S rRNA基因序列分析研究其多样性。【结果】嗜酸丝状放线菌的最佳分离方法为：土壤样品经分散差速离心预处理后,涂布添加了放线菌酮、制霉菌素和萘啶酮酸（各50 mg/L）的GTV培养基。用此方法共分离到放线菌369株,归为10个不同的颜色类群,其中6.6%为严格嗜酸放线菌,72.4%为中度嗜酸放线菌,21.0%为耐酸放线菌。52株嗜酸放线菌代表菌株分布于放线菌目中的12个属：链霉菌属(Streptomyces)、小单孢菌属(Micromonospora) 、诺卡氏菌属(Nocardia)、野野村菌属(Nonomuraea) 、韩国生工属(Kribbella) 、小双孢菌属(Microbispora)、马杜拉菌属(Actinomadura)、拟无枝菌酸菌属(Amycolatopsis)、指孢囊菌属(Dactylosporangium)、伦茨氏菌属(Lentzea)、游动四孢菌属(Planotetraspora) 和链嗜酸菌属(Streptacidiphilus),其中链霉菌分离菌株在系统发育树上形成12个不同的进化类群。【结论】所建立的选择性分离方法可用于土壤嗜酸丝状放线菌的高效分离;江西酸性土壤含有丰富多样的嗜酸丝状放线菌种属。     

Effects of dietary crude protein level on odour from pig manure

The objective of this study was to determine the effects of dietary crude protein (CP) level on odour emission, odour intensity, hedonic tone, and ammonia emission from pig manure and on manure composition (pH, total nitrogen, ammonium, volatile fatty acids, indolic, phenolic and sulphur-containing compounds). An experiment was conducted with growing pigs (n = 18) in a randomised complete-block design with three treatments in six blocks. Treatment groups were 12%, 15% and 18% CP diets. Barley was exchanged for soya-bean meal. Crystalline amino acids (AA) were included in the 12% CP diet up to the level of pigs' requirement; the same amount of AA was added to the 15% and 18% CP diets. Pigs with an initial body weight (BW) of 36.5 ± 3.4 kg (mean ± s.d.) were individually penned in partly slatted floor pens and offered a daily feed allowance of 2.8 × maintenance requirement for net energy (NE: 293 kJ/kg BW^0.75). Feed was mixed with water, 1/2.5 (w/w). Faeces and urine of each pig were accumulated together in a separate manure pit under the slatted floor. After an adaptation period of 2 weeks, the manure pits were cleaned and manure was collected. In the 5th week of the collection period, air samples for odour and ammonia analyses, and manure samples were collected directly from each manure pit. Air samples were analysed for odour concentration and for hedonic value and intensity above odour detection threshold. Manure samples were analysed for volatile fatty acids, and indolic, phenolic and sulphurous compounds, ammonium and total nitrogen concentrations. Reducing dietary CP from 18% to 12% lowered odour emission ( P < 0.05) and ammonia emission ( P = 0.01) from pig manure by 80% and 53%, respectively. Reduced dietary CP decreased total nitrogen, methyl sulphide, carbon disulphide, ethanethiol, phenol, 4-ethyl phenol, indole and 3-methyl indole concentrations in the manure ( P < 0.05). Volatile fatty acids and cresols concentrations in the manure of pigs fed different dietary CP levels were similar. A reduction of dietary CP and at the same time providing essential AA is an option to reduce odour emission as well as ammonia emission from pig manure.