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.     

Reliability of odour–genes covariance despite diet changes: a test in mound‐building mice

Odour–genes covariance (i.e. that individual odour similarities covary with genetic similarity) allows animals to assess their genetic relatedness to one another by comparing the similarities between their individual odours. The reliability of the odour–gene covariance phenomenon under environmental changes has not yet been explored. Using an innovative habituation‐generalization procedure, we tested whether the modifications of body odours induced by a change in diet interfered with the perceived similarity of individual odours of more genetically similar individuals. The effects of a change in diet, by adding aniseed flavour to the drinking water, was assessed in male mound‐building mice, a species in which odour–genes covariance has already been demonstrated. The results obtained indicate that mice perceive more similarities in the odours of sibling males than in those of nonrelatives, despite the effects of diet on body odours. We suggest that the reliability of signals arising from odour–genes covariance is likely a result of the independence between traits indicating genetic relatedness and indicator signals of biological states (e.g. diet) in body odours. © 2012 The Linnean Society of London, Biological Journal of the Linnean Society, 2012, 106 , 682–688.     

Close encounters: contributions of carbon dioxide and human skin odour to finding and landing on a host in Aedes aegypti

In a wind‐tunnel study, the upwind flight and source location of female Aedes aegypti to plumes of carbon dioxide (CO₂) gas and odour from human feet is tested. Both odour sources are presented singly and in combination. Flight upwind along the plumes is evident for both CO₂ and odour from human feet when the odours are presented alone. Similarly, both odour sources are located by more than 70% of mosquitoes in less than 3 min. When both CO₂ and odour from human feet are presented simultaneously in two different choice tests (with plumes superimposed or with plumes separated), there is no evidence that females orientate along the plume of CO₂ and only a few mosquitoes locate its source. Rather, the foot odour plume is navigated and the source of foot odour is located by over 80% of female Ae. aegypti. When a female is presented a plume of CO₂ within a broad plume of human foot odour of relatively low concentration, the source of CO₂ is not located; instead, flight is upwind in the diffuse plume of foot odour. Although upwind flight by Ae. aegypti at long range is presumably induced by CO₂ and the threshold of response to skin odours is lowered, our findings suggest that, once females have arrived near a prospective human host, upwind orientation and landing are largely governed by the suite of human odours, whereas orientation is no longer influenced by CO₂.     

ATP Modulates the Growth of Specific Microbial Strains

The regulatory function of extracellular ATP (exATP) in bacteria is unknown, but recent studies have demonstrated exATP induced enhanced secondary metabolite production and morphological differentiation in Streptomyces coelicolor. The growth of Streptomyces coelicolor, however, was unaffected by exATP, although changes in growth are common phenotypes. To identify bacteria whose growth is altered by exATP, we measured exATP-induced population changes in fast-growing microbes and actinomycetes in compost. Compared with the water-treated control, the addition of 10 ml 100 μM ATP to 10 g of compost enhanced the actinomycetes population by 30% and decreased fast-growing microbial numbers by 20%. Eight microbes from each group were selected from the most populated colony, based on appearance. Of the eight isolated fast-growing microbes, the 16S rRNA sequences of three isolates were similar to the plant pathogens Serratia proteamaculans and Sphingomonas melonis, and one was close to a human pathogen, Elizabethkingia meningoseptica. The growth of all fast-growing microbes was inhibited by ATP, which was confirmed in Pseudomonas syringae DC3000, a pathogenic plant bacterium. The growth of six of eight isolated actinomycetes strains, all of which were identified as close to Streptomyces neyagawaensis, was enhanced by ATP treatment. This study suggests that exATP regulates bacterial physiology and that the exATP response system is a target for the control of bacterial ecology.     

Production of lytic enzymes and siderophores, and inhibition of germination of basidiospores of Moniliophthora (ex Crinipellis) perniciosa by phylloplane actinomycetes

Streptomyces albovinaceus, Streptomyces caviscabies, Streptomyces griseus, Streptomyces setonii, and Streptomyces virginiae selected as antagonists of Moniliophthora (ex Crinipellis) perniciosa, the causal agent of cacao Witches’ broom, were examined in vitro to detect production of chitinases, β-1,3-glucanases, and cellulases. All the species produced chitinases, but not β-1,3-glucanases or cellulases, when grown on a liquid mineral medium containing glucose, colloidal chitin, or cell walls of M. perniciosa as a carbon source. There were no quantitative differences among species in the production of chitinase, however, the germination inhibition of basidiospores of M. perniciosa was higher when they were cultivated using glucose as a carbon source, followed by colloidal chitin and cell walls. All the species also produced hydroxymate type siderophores in similar quantities, and the quantity of siderophores did not correlate with the inhibition of basidiospore germination. The germination inhibition was more pronounced when S. albovinaceus, S. griseus, and S. virginiae were cultivated on iron-deficient medium, suggesting involvement of siderophores in the antagonism by these species of actinomycetes.     

Increased heterologous production of the antitumoral polyketide mithramycin A by engineered Streptomyces lividans TK24 strains

Mithramycin A is an antitumor compound used for treatment of several types of cancer including chronic and acute myeloid leukemia, testicular carcinoma, hypercalcemia and Paget’s disease. Selective modifications of this molecule by combinatorial biosynthesis and biocatalysis opened the possibility to produce mithramycin analogues with improved properties that are currently under preclinical development. The mithramycin A biosynthetic gene cluster from Streptomyces argillaceus ATCC12956 was cloned by transformation assisted recombination in Saccharomyces cerevisiae and heterologous expression in Streptomyces lividans TK24 was evaluated. Mithramycin A was efficiently produced by S. lividans TK24 under standard fermentation conditions. To improve the yield of heterologously produced mithramycin A, a collection of derivative strains of S. lividans TK24 were constructed by sequential deletion of known potentially interfering secondary metabolite gene clusters using a protocol based on the positive selection of double crossover events with blue pigment indigoidine-producing gene. Mithramycin A production was evaluated in these S. lividans strains and substantially improved mithramycin A production was observed depending on the deleted gene clusters. A collection of S. lividans strains suitable for heterologous expression of actinomycetes secondary metabolites were generated and efficient production of mithramycin A with yields close to 3 g/L, under the tested fermentation conditions was achieved using these optimized collection of strains.

     

西沙群岛海域海洋放线菌的分离及其抗菌活性

分离西沙群岛海域放线菌并研究其抗菌活性。采用干燥、辐射、冷冻及加热处理等11种样品预处理方式和10种培养基对海洋放线菌进行分离,对代表性菌株进行鉴定,并考察分离放线菌生长的海水依赖性。进一步以金黄色葡萄球菌、大肠埃希菌、啤酒酵母和扩展青霉为指示菌考察分离放线菌的抗菌活性。从西沙群岛海域样品中分离获得放线菌383株,其中专性海洋放线菌23株。选定93株代表菌株进行鉴定,93株菌隶属于9个科,11个属。不同培养基对分离放线菌菌株的数量及种类影响显著。6株放线菌对4种指示菌均有抑菌活性,其中4株为专性海洋放线菌,表明海洋环境具有丰富的放线菌资源,这些放线菌特别是专性海洋放线菌有望为新型抗菌物质的发现与开发提供菌种来源。     

Mutagenesis by ethidium bromide and N-methyl-N′-nitro-N-nitrosoguanidine in off-flavour compound producing strains ofStreptomyces

Various species of actinomycetes and cyanobacteria can impart earthy/musty off-flavours to drinking water supplies and to pond-raised fish and other aquatic food animals. The genetic determinants for production of the most common off-flavour compounds [geosmin and 2-methylisoborneol (MIB)] have not been extensively studied. An attempt has been rrlade to study the genetics of production of these compounds was demonstrated by DNA-curing analysis. The effects of two curing agents [ethidium bromide (EB) and N-methyl-N′-nitro-N-nitrosoguanidine (NTG)] on tha loss of linear plasmid DNA and generation of bald mutants (no aerial mycelia) inStreptomyces halstedii andStereptomyces violaceusniger which produce geosmin and MIB, respectively, were observed. Production of earthy/musty odour was not eliminated, but was reduced by 55–95% in the plasmid cured strain. Data suggested that off-flavour production is likely chromosomally-encoded in theseStreptomyces isolates.     

川楝内生放线菌的分离及多样性研究

【目的】分离四川省各个地区川楝内生放线菌并研究其物种多样性。【方法】应用7种选择性分离培养基分离样品根、茎、叶、树皮和果实中的内生放线菌,采用16SrRNA基因RFLP分析代表菌株多样性。【结果】研究共获得403株内生放线菌。不同地点、不同植株部位、不同培养基分离得到的内生放线菌数目均有差异。广元采集的样品分离得到的数目最多,为86株;最少的是绵阳,仅有12株。从植物表皮中分离到148株放线菌,占获得菌株总数的36.7%;而从果中分离到31株,仅占获得菌株总数的7.6%;虽然从根部分离到的数量也很少,但是其出菌率却是最高的。5号和3号培养基的分离效果最为理想。16S rRNA基因RFLP分析结果显示所有供试菌株在68%的相似性上聚在一起,在84%的相似水平上分成了10个遗传类型。代表菌株的16SrRNA基因序列测定及系统发育分析结果表明:分离得到的放线菌包括4个属,分别是链霉菌属(Streptomyces)、北里孢菌属(Kitasatospora)、节杆菌属(Arthrobacter)、克里布所菌属(Kribbella)。其中,链霉菌是优势类群,占代表菌株数目的比例高达91%,而稀有放线菌的比例只有9%。【结论】研究发现的川楝内生放线菌主要属于链霉菌属(Streptomyces)、北里孢菌属(Kitasatospora)、节杆菌属(Arthrobacter)、克里布所菌属(Kribbella)。     

The attractiveness of different odour sources from the fruit–host complex on Leptopilina boulardi, a larval parasitoid of frugivorous Drosophila spp.

In parasitoid insects, successful offspring development depends on the female’s ability to find a suitable host. Specific recognition is often based on responses to olfactory cues, but their source and nature have rarely been determined. –This paper deals with the recognition of odours involved in host location by Leptopilina boulardi[Barbotin, Carton & Kelner-Pillault] (Hymenoptera: Eucoilidae), a larval parasitoid of Drosophila species that develops in mature fruits. The nature and origin of volatile stimuli recognized among odours of the host–fruit complex, and the effect of learning on this recognition, were investigated. Oriented responses to these odours were observed in a four-armed olfactometer and were analysed with the observer software (Noldus Information Technology). Fruit odours alone (banana and pear) were not spontaneously attractive to naive parasitoids, whereas naturally-infested bananas were highly attractive. The attraction was related to the odour that adult Drosophila left on the substrate but not to Drosophila oviposition activity or larval development. A synergism between some fruit odours (banana and pear) and the odour left by adult Drosophila on damp filter paper was observed. However, when testing a non-fruit substrate (mushroom), no synergism was observed. Thus, female L. boulardi may innately recognize host–food substrate odours associated with odours from the adult stage of their host. In addition, an oviposition experience on an infested banana allows L. boulardi females to memorise the fruit odour itself through associative learning. The adaptive significance of this process is discussed.     

西沙海藻共附生放线菌的分离鉴定及其抗菌活性评价

【目的】为了探究南海海藻共附生放线菌资源的多样性及潜在的应用价值,对中国西沙群岛来源的海藻进行共附生放线菌的分离鉴定与抗菌活性筛选。【方法】利用稀释涂布平板法,采用2种不同分离培养基对不同采样位点的6种海藻进行放线菌分离;通过16S rRNA基因序列分析、构建系统发育树对分离的放线菌进行鉴定;用打孔法对无乳链球菌(Streptococcus agalactiae)等10种敏感细菌进行抗菌活性筛选;对筛选得到的目标活性菌株HZ014进行全基因组测序,通过AntiSMASH在线工具分析其次级代谢产物生物合成基因簇,预测其产生新型活性物质的潜力。【结果】从6种海藻中分离得到36株共附生放线菌,基于16S rRNA基因序列比对和系统发育分析,鉴定结果为链霉菌属(Streptomyces) 2株、红球菌属(Rhodococcus) 2株、诺卡氏菌属(Nocardia)3株、小单孢菌属(Micromonospora) 5株和盐孢菌属(Salinispora) 24株;抗菌活性筛选结果表明,36株共附生放线菌发酵粗提物对至少1种敏感细菌表现出一定的抑制作用,不同菌株发酵粗提物的抗菌活性存在明显差异,...     

Characterization of actinomycetes isolated from the indoor air of the church of Saint Katherine Monastery, Egypt

Actinomycetes were isolated from the indoor air of the church of Saint Katherin Monastery during different visiting hours. Fifty air samples collected over one year by using automated air sampler were plated on four different media. The low nutrient-content medium 1/10 SC was significantly effective in recovery of actinomycetes compared to the other formula of rich media. Average counts of bacteria, actinomycetes and fungi were 487, 65 and 90 cfu/m³, respectively. Fifty-six morphologically different actinomycetes isolates were recovered during this study assigned into five different genera, in addition to three unidentified isolates. Tentative identification of the isolates indicated predominance of genus Streptomyces, representing 59% of the isolates. Isolates were screened for resistance to 11 antibiotics, antimicrobial activities towards seven microbial strains, Growth on 12 different carbon source, acid production and pigmentation. About 77% of actinomycete isolates were resistant to the antibiotics with different resistance patterns. 12.5% of the airborne actinomycetes exhibited antinmicrobial activities. The isolates showed wide variation in carbon source usage. Forty percent of the isolates were able to utilize all tested carbon sources while 80% were acid producers. Melanin production was produced by 34% of the isolates. While 45% of the isolates were able to produce other diffusible pigments, the majority of the pigments were brown in color however; yellow, orange and green pigments were produced by a number of isolates. The impact of these activities on the historical objects of the church and the tourist’s health is discussed.     

Foraging behavioural changes induced by conspecific and heterosubspecific odours in two strains of wild mice

Mice in wild populations of the two subspecies Mus musculus domesticus and Mus musculus musculus may potentially compete for food. Because of the importance of olfaction in mice, we hypothesised that the presence of unfamiliar conspecific or heterosubspecific chemical cues could play a role in access to and use of food resources. We used an experimental design that tested this assumption with males from two strains, originated from wild populations of these subspecies, as subjects. Exploratory activity, latency of food approach, time and frequency on the food area, number of seeds eaten and foraging rate (number of seeds eaten/time on the food areax100) were compared for three different categories of odours (own, same strain and other strain odours) in both strains. In a foraging context, unfamiliar odours induced behavioural changes in male mice, especially an increase in exploratory activity from the more (same strain) to the less similar odour (different strain), and a reduction of time spent in the food area. Odour similarity related to genetic proximity in Mus and the cost-benefit ratio of an encounter are two possible explanations for the different processes involved in the treatment of odours in these two strains of mice.     

A chitinolytic actinomycete complex in black soil

A chitinolytic actinomycete complex in chernozem soil has a specific taxonomic composition, which differs from that of the actinomycete complex which is 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 to 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 that are grown in the nutrient media with chitin.     

Divergent responses of two cereal aphids to previous infestation of their host plant

The role of pollen odour in resource location by the pollen beetle, Meligethes aeneus (Fabricius) (Coleoptera: Nitidulidae), a pollen-feeding insect regarded as a pest of oilseed rape, Brassica napus L., (Brassicaceae) crops, was investigated in a linear track olfactometer. Both male and female beetles were attracted to the odour of whole oilseed rape flowers, indicating that these insects can locate their host plants using floral odours as cues. The attractive odour of flowers was found to emanate from all floral parts tested: the petals/sepals, the anthers, and from pollen itself. Therefore, at least part of the attractive odour of oilseed rape flowers emanates from pollen. Beetles were more attracted to floral samples containing anthers than those without anthers when these odours were directly compared in a choice-test, and this indicates that there were detectable differences between them. Anthers and pollen may therefore release distinctive odours that are quantitatively and/or qualitatively different from the odour of the rest of the flower. These experiments support the hypothesis that pollen-seeking insects use pollen odour cues to locate this food source.     

Lakewide odours in Ontario and New Hampshire caused by Chrysochromulina breviturrita Nich. (Prymnesiophyceae)

At densities of 500–9 000 cells/ml, the planktonic alga Chrysochromulina breviturrita Nich. produced obnoxious lake-wide odours in five lakes between 1978 and 1980. In one case, CuSO₄ was used to kill the organism and control the odour, but in the other four lakes, the species persisted for 4–6 weeks before populations declined and odours disappeared. This is the first recorded instance of odour production by any member of the Prymnesiophyceae.     

An olfactory recognition system in the ferret Mustela furo L. (Carnivora: Mustelidae)

This study demonstrates that ferrets can use variations in odours from anal sac secretions as a communication system. Odour preference tests showed that ferrets can discriminate between male and female ferret anal sac odours, between strange and familiar, familiar and their own, and fresh and 1-day-old odours. They did not discriminate between fresh and 2-h-old odours, nor did male ferrets discriminate between the odours of oestrous and anoestrous females. Ferrets were more attracted to the odours of the opposite sex than to those of their own sex. When faced by an opponent, male ferrets were more aggressive in the presence of their own rather than their opponent's odour, and less aggressive with their opponent's odour than with that of a known, dominant animal's odour. These results are consistent with both a sex attraction role and a territorial defence role for anal sac odours. A scent-matching mechanism for territorial defence is supported, although a neighbour-neighbour recognition/avoidance mechanism cannot be rejected. Gas chromatography revealed sexually and individually distinct profiles of volatile compounds in anal sac extracts, but no consistent seasonal trends. Females had high concentrations of 2,3-dimethylthietane and/or 3,4-dimethyl-1,2-dithiolane. Males usually had high concentrations of indole. 2-Propylthietane was an important constituent in most individuals. These differences in concentration were significant and could provide an olfactory recognition system of sex and individual identity.     

Diversity of Aquatic Actinomycetes in Lakes of the Middle Plateau, Yunnan, China

A total of 749 sediment and water samples were collected from 12 lakes of the Middle Plateau of Yunnan from 1983 to 1993. The diversity and biological characteristics of the aquatic actinomycetes in these lakes were studied. Sixteen genera of actinomycetes were isolated from these samples. Micromonospores assumed a notable dominance (from 39 to 89%) in the actinomycete populations of these lake sediments. Streptomycetes were the second most abundant organisms. The diversity and counts of actinomycetes varied with the season. Thermophilic actinomycetes have a wide distribution in these lakes, but their counts were smaller. The cell wall compositions of certain Micromonospora and Streptomyces strains from an alkaline lake revealed an unusual combination of glycine and isomers of diaminopimelic acid. It seems that aquatic actinomycetes play a significant role in the decomposition of organic substances, including some toxic compounds such as phenol, in these lakes. It also appears that aquatic actinomycetes are one of the important resources for screening useful enzymes and metabolites.     

Differential attraction of malaria mosquitoes to volatile blends produced by human skin bacteria

The malaria mosquito Anopheles gambiae sensu stricto is mainly guided by human odour components to find its blood host. Skin bacteria play an important role in the production of human body odour and when grown in vitro, skin bacteria produce volatiles that are attractive to A. gambiae. The role of single skin bacterial species in the production of volatiles that mediate the host-seeking behaviour of mosquitoes has remained largely unknown and is the subject of the present study. Headspace samples were taken to identify volatiles that mediate this behaviour. These volatiles could be used as mosquito attractants or repellents. Five commonly occurring species of skin bacteria were tested in an olfactometer for the production of volatiles that attract A. gambiae. Odour blends produced by some bacterial species were more attractive than blends produced by other species. In contrast to odours from the other bacterial species tested, odours produced by Pseudomonas aeruginosa were not attractive to A. gambiae. Headspace analysis of bacterial volatiles in combination with behavioural assays led to the identification of six compounds that elicited a behavioural effect in A. gambiae. Our results provide, to our knowledge, the first evidence for a role of selected bacterial species, common on the human skin, in determining the attractiveness of humans to malaria mosquitoes. This information will be used in the further development of a blend of semiochemicals for the manipulation of mosquito behaviour.     

Perceptual and neural olfactory similarity in honeybees

The question of whether or not neural activity patterns recorded in the olfactory centres of the brain correspond to olfactory perceptual measures remains unanswered. To address this question, we studied olfaction in honeybees Apis mellifera using the olfactory conditioning of the proboscis extension response. We conditioned bees to odours and tested generalisation responses to different odours. Sixteen odours were used, which varied both in their functional group (primary and secondary alcohols, aldehydes and ketones) and in their carbon-chain length (from six to nine carbons).The results obtained by presentation of a total of 16 × 16 odour pairs show that (i) all odorants presented could be learned, although acquisition was lower for short-chain ketones; (ii) generalisation varied depending both on the functional group and the carbon-chain length of odours trained; higher generalisation was found between long-chain than between short-chain molecules and between groups such as primary and secondary alcohols; (iii) for some odour pairs, cross-generalisation between odorants was asymmetric; (iv) a putative olfactory space could be defined for the honeybee with functional group and carbon-chain length as inner dimensions; (v) perceptual distances in such a space correlate well with physiological distances determined from optophysiological recordings of antennal lobe activity. We conclude that functional group and carbon-chain length are inner dimensions of the honeybee olfactory space and that neural activity in the antennal lobe reflects the perceptual quality of odours.