An airborne actinobacteria Nocardiopsis alba isolated from bioaerosol of a mushroom compost facility

Actinobacteria are widely distributed in many environments and represent the most important trigger to the occupant respiratory health. Health complaints, including hypersensitivity pneumonitis of the workers, were recorded in a mushroom compost facility (MCF). The studies on the airborne bacteria were carried out to find a possible microbiological source of these symptoms. Culture analysis of compost bioaerosols collected in different location of the MCF was performed. An assessment of the indoor microbial exposure revealed bacterial flora of bioaerosol in the mushroom compost facility represented by Bacillus, Geobacillus, Micrococcus, Pseudomonas, Staphylococcus spp., and actinobacterial strain with white aerial mycelium. The thermotolerant actinobacterial strain of the same morphology was repeatedly isolated from many locations in MCF: air, compost sample, and solid surface in production hall. On the base of complex morphological, chemotaxonomic, and phylogenetic characteristics, the isolate has been classified as Nocardiopsis alba. Dominant position of N. alba in microbial environment of the mushroom compost facility may represent an indicator microorganism in compost bioaerosol. The bioavailability of N. alba in mushroom compost facility creates potential risk for the health of workers, and the protection of respiratory tract and/or skin is strongly recommended.     

<Emphasis Type="Italic">Nocardiopsis yanglingensis</Emphasis> sp. nov., a thermophilic strain isolated from a compost of button mushrooms

A strain named A18 was recovered from a compost of button mushrooms. It was characterized using a polyphasic approach. On the basis of 16S rRNA gene sequence comparison, it belonged to the genus Nocardiopsis and was most closely related to the type strains of Nocardiopsis flavescens (sequence similarity 98.0%), Nocardiopsis prasina (97.5%), Nocardiopsis metallicus (97.4%), Nocardiopsis alba (97.3%). The combination of phylogenetic analysis, DNA–DNA hybridization, phenotypic characteristics and chemotaxonomic data supported the proposal that strain A18 represents a new species of the genus Nocardiopsis, for which the name Nocardiopsis yanglingensis sp. nov. was proposed (type strain A18^T = KCTC 19723^T = CCTCC 209063^T).     

Production and characterization of lipopeptide biosurfactant by a sponge-associated marine actinomycetes <Emphasis Type="Italic">Nocardiopsis alba</Emphasis> MSA10

A sponge-associated marine actinomycetes Nocardiopsis alba MSA10 was screened and evaluated for the production of biosurfactant. Biosurfactant production was confirmed by conventional screening methods including hemolytic activity, drop collapsing test, oil displacement method, lipase production and emulsification index. The active compound was extracted with three solvents including ethyl acetate, diethyl ether and dichloromethane. The diethyl ether extract was fractionated by TLC and semi-preparative HPLC to isolate the pure compound. In TLC, a single discrete spot was obtained with the R _f 0.60 and it was extrapolated as valine. Based on the chemical characterization, the active compound was partially confirmed as lipopeptide. The optimum production was attained at pH 7, temperature 30°C, and 1% salinity with glucose and peptone supplementation as carbon and nitrogen sources, respectively. Considering the biosurfactant production potential of N. alba, the strain could be developed for large-scale production of lipopeptide biosurfactant.     

New structures and composition of cell wall teichoic acids from Nocardiopsis synnemataformans,Nocardiopsis halotolerans,Nocardiopsis composta and Nocardiopsis metallicus: a chemotaxonomic value

The structures of the cell wall teichoic acids (TA) from some species of the genus Nocardiopsis were established by chemical and NMR spectroscopic methods. The cell walls of Nocardiopsis synnemataformans VKM Ac-2518^T and Nocardiopsis halotolerans VKM Ac-2519^T both contain two TA with unique structures—poly(polyol phosphate-glycosylpolyol phosphate)—belonging to the type IV TA. In both organisms, the minor TA have identical structures: poly(glycerol phosphate-N-acetyl-β-galactosaminylglycerol phosphate) with the phosphodiester bond between C-3 of glycerol and C-4 of the amino sugar. This structure is found for the first time. The major TA of N. halotolerans has a hitherto unknown structure: poly(glycerol phosphate-N-acetyl-β-galactosaminylglycerol phosphate), the N-acetyl-β-galactosamine being acetalated with pyruvic acid at positions 4 and 6. The major TA of N. synnemataformans is a poly(glycerol phosphate-N-acetyl-β-galactosaminylglycerol phosphate) with the phosphodiester bond between C-3 of glycerol and C-3 of the amino sugar. The cell walls of Nocardiopsis composta VKM Ac-2520 and N. composta VKM Ac-2521^T contain only one TA, namely 1,3-poly(glycerol phosphate) partially substituted with N-acetyl-α-glucosamine. The cell wall of Nocardiopsis metallicus VKM Ac-2522^T contains two TA. The major TA is 1,5-poly(ribitol phosphate), each ribitol unit carrying a pyruvate ketal group at positions 2 and 4. The structure of the minor TA is the same as that of N. composta. The results presented correlate well with the phylogenetic grouping of strains and confirm the species and strain specific features of cell wall TA in members of the genus Nocardiopsis.     

Systematic studies on the Zygophyllaceae of Saudi Arabia: A new variety and new variety combination in Tetraena

During 2013–2014, fresh material of Tetraena alba was collected from two localities in Saudi Arabia. Detailed morphological examination of these specimens revealed that there is a new variety of this species, for which the name T. alba var. arabica is proposed. Fresh material of Zygophyllum amblyocarpum was also collected from a different locality, and based on its characteristics is transferred to Tetraena with the new combination T. alba var. amblyocarpa. T. alba and its varieties are illustrated and morphological characters supplied to differentiate among them.     

Proteolytic activity in Plagiodera versicolora Laicharting (Coleoptera: Chrysomelidae): Characterization of digestive proteases and effect of host plants

Proteolytic profiles in the midgut of Plagiodera versicolora were studied using biochemical approaches, and the effects of host plants on possible changes in their activity were determined. Morphology of the alimentary canal revealed several areas of sections, namely bucca, pharynx, esophagus, crop, midgut, ileum, rectum and anus. A pH of 6 and 11 was found to be optimal for soluble and membrane-bound fractions, by using azocasein 2% as a substrate. Determination of specific proteases demonstrates the presence of trypsin-like, chymotrypsin-like, elastase, cathepsin B, cathepsin L and cathepsin D, as well as two exopeptidases. Regarding site of activity for each specific protease, it was found that the major activity of cathepsin B and cathepsin L was in the soluble fraction, chymotrypsin, cathepsin D and two exopeptidases in membrane-bound fraction. Additionally, trypsin-like and elastase activities had no significant differences between fractions. The presence of the above mentioned specific proteases was verified using the specific inhibitors PMSF, TLCK, TPCK, cystatin, phenanthroline and DTT. Feeding of the beetle on four host plants: including Salix aegyptica, S. alba, Populus alba and P. caspica, from the 1st larval instar to adult, revealed the highest trypsin-, chymotrypsin-like and elastase activities in the individuals fed on S. aegyptica and S. alba, respectively. Regarding cathepsins B and L, the highest activities were observed on S. alba and S. aegypticum but cathepsin D was higher in S. Alba and P. alba. Feeding on S. alba and S. aegypticum showed the highest activities of amino- and carboxy-peptidases, respectively.     

Halophilic and salts tolerant protoplast cultures of mangrove plants, Sonneratia alba and Avicennia alba

The effects of sea salts, NaCl, KCl, MgCl₂, MgSO₄, and CaCl₂, on the growth of protoplast cultures of two mangrove species, Sonneratia alba and Avicennia alba, were investigated using 96-well culture plates. Plants of these two species naturally grow at the seaward side of a mangrove forest. Cotyledon protoplasts of S. alba showed halophilic nature to NaCl, KCl, and MgCl₂ at low concentrations (10–50 mM) when cultured in Murashige and Skoog’s (MS) medium containing 0.6 M mannitol. CaCl₂ at a concentration higher than 25 mM was inhibitory to cell growth. On the other hand, in protoplast culture of A. alba suspension cells, which were induced from cotyledon tissues, in the modified amino acid (mAA) medium containing 1.2 M sorbitol, tolerance to NaCl, MgCl₂ and MgSO₄ were observed at a wide range of concentrations up to 400 mM. CaCl₂ was always inhibitory for cell divisions in A. alba, but stimulatory for spherical enlargement of cells. However, no difference in cell enlargement was observed among other salts. Similarity and difference in reactivity to salts between protoplasts and suspension cells from our previous studies were discussed in relation to the site of salt tolerance or halophilic adaptation within mangrove cells. For protoplast cultures, the site(s) for response of S. alba and A. alba are located in the cytoplasm and/or the cell membrane.     

Novel marine Nocardiopsis dassonvillei-DS013 mediated silver nanoparticles characterization and its bactericidal potential against clinical isolates

The sediment marine samples were obtained from several places along the coastline of the Tuticorin shoreline, Tamil Nadu, India were separated for the presence of bioactive compound producing actinobacteria. The actinobacterial strain was subjected to 16Sr RNA sequence cluster analysis and identified as Nocardiopsis dassonvillei- DS013 NCBI accession number: KM098151. Bacterial mediated synthesis of nanoparticles gaining research attention owing its wide applications in nonmedical biotechnology. In the current study, a single step eco-friendly silver nanoparticles (AgNPs) were synthesized from novel actinobacteria Nocardiopsis dassonvillei- DS013 has been attempted. The actinobacterial mediated silver nanoparticles were characterized by TEM, UV–Visible, XRD, FT-IR spectroscopy. The initial detection of AgNPs was identified using UV–Vis spectrum and confirmed by the appearance of absorbance peak at 408 nm. A Fourier transform infrared spectroscopy (FT-IR) result reveals the presence of protein component in the culture supernatant may act as protecting agents. The XRD pattern indicated that the typical peaks reveal the presence of nanoparticles. The TEM morphology confirms the formation of circular and non uniform distributions of AgNPs with the size range from 30 to 80 nm. The antibacterial activity of both isolated actinobacterial (IA) and silver nanoparticles mediated actinobacterial (SNA) of Nocardiopsis dassonvillei- DS013 were done by well diffusion method against selected clinical isolates of bacteria, namely Escherichia coli, Enterococcus sp., Pseudomonas sp., Klebsiella sp., Proteus sp., Shigella sp., Bacillus subtilis, and Streptococcus sp. When compared to isolated actinobacteria, the SNA shows the better antibacterial activity against clinical isolates.     

Molecular phylogenetic analyses based on the nuclear rRNA genes and the intron–exon structures of the nuSSU rRNA gene in Dictyocatenulata alba (anamorphic Ascomycota)

Molecular phylogenies inferred from the nuclear small subunit rRNA gene (nuSSU), nuclear large subunit rRNA gene D1/D2 region (nuLSU), and ITS-5.8S rRNA gene (ITS) indicated that five cultures of the lichenized hyphomycete Dictyocatenulata alba isolated from Japan form a monophyletic clade with high bootstrap support, and a close relationship to the Ostropomycetidae (Lecanoromycetes, Pezizomycotina, Ascomycota). Insertion sequences were found in the nuSSU of all isolates [e.g., nine insertions in the strain JCM 5358 (Japan Collection of Microorganisms)], some of which were group I introns. Five new insertion positions were found among the D. alba isolates. Using BLAST, none of the insertion sequences of D. alba were closely related to those of fungi or other organisms deposited in public DNA databases. Insertion positions were similar in some isolates, and two positions were common to all isolates. Although all phylogenetic analyses based on nuSSU, nuLSU, and ITS revealed the monophyly of D. alba, the isolates were divided into two (in the nuSSU tree) or three (in the nuLSU and ITS trees) groups. Based on the phylogenetic analyses and the intron–exon structures, the five isolates identified as D. alba belong to three cryptic species and therefore D. alba should be considered a species complex. The very slow-growing, tough agar colonies of the isolates, the occurrence of the species on both slightly lichenized and nonlichenized surfaces of trees, or pebbles (rarely on soil), suggest that the members of the D. alba complex may be lichenized. The photobiont was not clearly identified in this study.     

Predation-risk effects of predator identity on the foraging behaviors of frugivorous bats

Predators directly and indirectly affect the density and the behavior of prey. These effects may potentially cascade down to lower trophic levels. In this study, we tested the effects of predator calls (playbacks of bird vocalizations: Tyto alba, Speotyto cunicularia, and Vanellus chilensis), predator visual stimuli (stuffed birds) and interactions of visual and auditory cues, on the behavior of frugivore phyllostomid bats in the field. In addition, we tested if the effects of predation risk cascade down to other trophic levels by measuring rates of seed dispersal of the tree Muntingia calabura. Using video recording, we found that bats significantly decreased the foraging frequency on trees when a visual cue of T. alba was present. However, no stimuli of potential predatory birds, including vocalization of T. alba, affected bat foraging frequency. There was a change in bat behavior during 7 min, but then their frequency of activity gradually increased. Consequently, the presence of T. alba decreased by up to ten times the rate of seed removal. These results indicate that risk sensitivity of frugivorous phyllostomid bats depends on predator identity and presence. Among the predators used in this study, only T. alba is an effective bat predator in the Neotropics. Sound stimuli of T. alba seem not to be a cue of predation risk, possibly because their vocalizations are used only for intraspecific communication. This study emphasizes the importance of evaluating different predator stimuli on the behavior of vertebrates, as well as the effects of these stimuli on trait-mediated trophic cascades.     

Artificial hybridization of some Abies species

Crossability relationships between six species of the Mediterranean, North American and Asian firs was tested using Abies alba and A. nordmanniana as female parents and A. alba, A. numidica, A. procera, A. grandis, and A. holophylla as pollen parents. An overwhelming majority of the crosses attempted was found to be compatible. In particular, it is true of the A. alba cross with A. numidica and those of A. nordmanniana with A. alba, A. numidica, A. procera, and A. holophylla. The crossing A. nordmanniana × A. grandis was the only exception producing empty seeds. Cytological study revealed the gametophytic incompatibility to be responsible for reproductive isolation of these species. At seedling level, all the interspecific crosses of A. nordmanniana surpassed in height growth self-pollinated control. The cross A. alba × A. numidica was comparable in this respect with control variants from open and self-pollination. Except for height growth, some characteristics of needle stomata are provided for individual crosses. The crosses A. nordmanniana with A. procera and A. holophylla represent unique interspecific combinations whose existence has not been reported yet. Based on needle stomata characteristics, the potential for increased resistance and drought tolerance of the hybrids with A. numidica involved as parental species is discussed.     

Extensive Clonal Assemblies in Populus alba and Populus x canescens from the Iberian Peninsula

Riparian vegetation in temperate zones holds great environmental importance and thus its conservation should be a priority. Among riverine tree species, genus Populus stands out, being also a model organism in research. In this work, we present a genetic analysis of the hybrid Populus x canescens and its parent species Populus alba and Populus tremula, with special emphasis in P. alba with which backcrosses frequently occur. This study focuses on the river Douro basin (Spain) where the presence of hybrids has been previously reported. Nuclear microsatellite markers and Bayesian statistical analysis have been used for the detection of hybrids and purebred stands. This methodology has also made possible the study of clonality in the taxonomic continuum P. alba–P. x canescens. Our results highlight the existence of a small number of genotypes accounting for most of the individual trees and stands. Possible causes leading to this situation are discussed. Also, the presence of individuals genetically close to ornamental cultivars is reported for the first time. Finally, concern about the present situation of P. alba genetic resources is brought up.     

Phylogeography of Populus alba (L.) and Populus tremula (L.) in Central Europe: secondary contact and hybridisation during recolonisation from disconnected refugia

The central aim of this paper is to clarify the picture of postglacial recolonisation and the reconstruction of refugia of Populus alba (L.) and Populus tremula (L.) in the light of hybridisation of the two species. We focussed our study on Central and Southeastern Europe including reference samples from Spain, Sweden and Northern Africa.We investigated 414 individuals of 26 populations using restriction fragment length polymorphisms (PCR-RFLPs) in six maternally inherited chloroplast markers. Altogether, 57 haplotypes were analysed of which four indicated hybridisation events in the past. Phylogeographic structure was found for P. alba with low diversity in Eastern Europe versus high diversity in Italy and Central Europe. A lack of phylogeographic structure was assessed for P. tremula as expected for a boreal forest tree, and diversity was evenly distributed in the studied populations. Two main refugia were identified for P. alba in Italy and Romania. A previously described hybrid zone between species in Central Europe turned out also to be a zone of contact between southern and eastern chloroplast lineages in P. alba. In contrast, P. tremula recolonised its present habitats in Central Europe from several refugia near the former ice cap. We assume separate disconnected refugia for P. alba and P. tremula and suggest an immigration scenario involving the mixing of colonisation routes and interspecific introgression to be responsible for the observed patterns.     

To be or not to be native: From the discourse on the official status of Salix alba to the recommendation to strengthen the protection of marginal populations in Latvia,with notes to related taxa

In Europe, Salix euxina is considered to be an adventive species, while S. alba is regarded as native. When assessing the protection of species in wild habitats, it is important to know the status of each species so that not to protect unreasonably alien species. Analysing the sources of literature, it was noticed that the views of Latvian authors differed — both on the status of the indigenous S. alba, as well as the adventive S. euxina. Research has shown that the ‘official status’ assigned to both basic species on the national level differs and contradicts the scientifically recognized status. This is an interesting case when certain state institutions responsible for nature protection have officially recognized a native species as an alien one and an alien species as a native one through laws and regulations. Considering that there are marginal populations of S. alba of the natural distribution range in Latvia, the most valuable of which occur in the basin of the River Gauja, the current possible protection of S. alba on the national level was analysed. Analysing the legislation, it has been established that rare habitats with S. alba woods (Salicetum albae) on alluvial soils of river flats are also included among the specially protected habitats. Analysing the individual protection regulations of the two largest specially protected areas, i. e. the Protected Landscape Area of the Northern Gauja and the Gauja National Park, it has been concluded that the current protection regime in the country does not ensure full protection of marginal populations of S. alba in natural habitats. This is because the individual protection regulations do not include the specific protected habitats of Salicetum albae and allow the felling of young willows, which contradicts the provisions of the legal act on the structural elements of the habitat Salicetum albae. In order to ensure full protection of marginal populations of S. alba and special habitats formed by the species in Latvia, it is necessary to update and define more accurately the laws and regulation on its protection.     

Variation in terpene composition of Artemisia alba in relation to environmental conditions

The essential oil from inflorescences of Artemisia alba Turra (Compositae) was analysed by GLC; 33 different components were detected, 21 of which were identified by GC-MS. The composition of the extract changes with environmental and climatic conditions of the sites of collection (Adriatic area-NE Italy). Artemisia alba produces mainly sesquiterpene hydrocarbons when growing in thermophytic plant communities and mostly oxygenated monoterpenes in mesophytic plant communities. The results of ordination and cluster analysis performed on chemical data allowed the identification of four chemical types of A. alba.     

The nature of polyploidy inReseda sect.Leucoreseda (Resedaceae)

We have found two chromosome levels (n = 10 and n = 20) in Spanish species ofReseda sect.Leucoreseda, four species on the first (R. undata, R. paui, R. suffruticosa, R. barrelieri), only one on the second level (R. alba). As already the species with n = 10 apparently behave as polyploids, we propose x = 5 as the original basic chromosome number for this section. The seed protein profile ofR. alba (2n = 40) reveals close relationships withR. undata (2n = 20), while the esterase isozymes suggest affinities withR. paui (2n = 20). Thus,R. alba can be regarded either as an autopolyploid fromR. paui or more likely as an allopolyploid fromR. paui andR. undata. Finally, the DNA values suggest a diploidization process inR. alba since its origin.     

Morphological Characters Useful for the Delimitation of Taxa Within Viola Subsect. Viola (Violaceae): A Morphometric Study from the West Carpathians

Forty-nine morphological characters were scored or measured on 44 populations (376 individuals) of Viola subsect. Viola from the West Carpathians and adjacent areas (Slovakia, Czech Republic, Austria and Hungary). The presence of six species, namely V. alba (represented by subsp. alba), V. ambigua, V. collina, V. hirta, V. odorata and V. suavis s.l. was revealed based on pollen fertility, cytological and morphometric analyses. The morphological characters traditionally used to delimit taxa within the subsection and those revealed by our study as most reliable are widely discussed. A key for identifying the taxa and most common hybrids of subsection Viola occurring in the West Carpathians is presented. Chromosome counting and flow cytometry were used to determine the ploidy levels of the populations studied. All individuals of V. alba subsp. alba, V. collina, V. hirta and V. odorata were tetraploid, while those of V. ambigua and V. suavis s.l. were octoploid.     

Early physiological responses of Abies alba and Rubus fruticosus to ungulate herbivory

For the last 50 years, Chamois (Rupicapra rupicapra) population exponentially increased in the French Alps. This herbivore faces a food shortage in the winter; its diet mainly includes woody species (Abies alba Mill.) and brambles (Rubus fruticosus). The physiological response following simulated herbivory of these two plants was assessed. More precisely, the effects of leaf clipping and ungulate saliva application on the epidermal flavonoids and chlorophyll contents were considered as an induced structural response. The chlorophyll fluorescence (Fv/Fm) was also measured to provide information on the photosynthesis status as an induced functional response. The non-invasive techniques used in this experiment highlighted an induced response, emerging during the days following the simulated herbivory. R. fruticosus was drastically affected, photosynthesis was disturbed (decreased Fv/Fm) and mortality began on day 5 reaching 100 % less than 1 week later. Physiological parameters were also affected in A. alba (slight decrease of Fv/Fm and lower content of flavonoids), but this species recovered and survived no matter what the stressing treatment was. These results did not point out an efficient repulsive-induced response of A. alba and R. fruticosus to herbivory. The existence of constitutive defences such as prickles (R. fruticosus) or low-digestible tannins (A. alba) is no more efficient to escape from ungulates consumption. Nevertheless, in spite of the increase of the ungulates exponential demography, these two plants do not face rarefaction problem in the French Alps. Thus, survival of A. alba stands is probably linked to the recovery ability of saplings, when R. fruticosus stands maintain themselves thanks to the strong resprouting capacity of this species.