Genomic Markers for Differentiation of Francisella tularensis subsp. tularensis A.I and A.II Strains

Tularemia is caused by two subspecies of Francisella tularensis, F. tularensis subsp. tularensis (type A) and F. tularensis subsp. holarctica (type B). F. tularensis subsp. tularensis is further subdivided into two genetically distinct populations (A.I and A.II) that differ with respect to geographical location, anatomical source of recovered isolates, and disease outcome. Using two human clinical isolates, suppression subtractive hybridization was performed to identify 13 genomic regions of difference between A.I and A.II strains. Two PCR assays, one to identify A.I and A.II as well as to discriminate between F. tularensis subsp. holarctica and F. novicida and another specific for A.I, were developed. This is the first report to identify and characterize conserved genomic differences between A.I and A.II.     

Taxonomy and Morphology of Axonchium (Nematoda: Belondiroidea), and a description of A. thornei n. sp.

Useful diagnostic characters in the nematode genus A xonchium include: lip shape, styler length, shape of the esophageal constriction, presence or absence of spiral musculature in the esophageal sheath, proportion of the esophageal length occupied by the esophageal expansion, length and shape of cardia, shape of the vulva and vaginal cuticularization, development of the anterior gonad, shape of the posterior uterus, subcuticle thickness at mid-body, tail shape, number and arrangement of supplements and caudal pores, and body measurements. A. thornei n. sp. is separated from A. choristum by its thinner subcuticle at mid-body, number of supplements, and shorter spicules, from A. solitare by presence of males, and from both species by the female tail shape and shorter stylet. A. saccatum is synonymized with A. gossypii and A. nitidum is synonymized with A. bulbosum. A. leptocephalum, A. Iongicollis, A. magnicollis, and A. tenuicollis are made species inquirendae. A key to 25 species of Axonchium is given.     

Reclassification of Aeromonas hydrophila subsp. dhakensis Huys et al. 2002 and Aeromonas aquariorum Martínez-Murcia et al. 2008 as Aeromonas dhakensis sp. nov. comb nov. and emendation of the species Aeromonas hydrophila

Previous studies indicate that Aeromonas aquariorum and Aeromonas hydrophila subsp. dhakensis are the same taxon and suggest that they should be synonymized. Using a polyphasic approach, the phenotypic and phylogenetic relationship of A. aquariorum with the 3 defined A. hydrophila subspecies (i.e. dhakensis, hydrophila, ranae) was investigated. Phylogenetic trees derived from the 16S rRNA, rpoD or gyrB genes and a multilocus phylogenetic analysis (with the concatenated sequences of gyrB, rpoD, recA, dnaJ and gyrA) confirmed that both A. aquariorum and A. hydrophila subsp. dhakensis are a unique taxon, different from the other A. hydrophila subspecies, corroborating the phenotypic and DNA–DNA hybridization (DDH) results. A formal synonymization of A. aquariorum and A. hydrophila subsp. dhakensis and a reclassification of both as Aeromonas dhakensis sp. nov. comb nov. is therefore proposed.     

Comparative chemical composition of Agastache mexicana subsp. mexicana and A. mexicana subsp. xolocotziana

A comparative chemical analysis of Agastache mexicana subsp. mexicana and A. mexicana subsp. xolocotziana reveals that their methanol extracts constituents were very similar, with acacetin and (2-acetyl)-7-O-glucosyl acacetin being the most abundant compounds obtained. These results are consistent with the information reported for other species of Agastache. However, GS-MS analyses showed that methyl chavicol, limonene and linalool were the main constituents of the essential oils of A. mexicana subsp. mexicana, while pulegone, menthone and isopulegone were the major constituents found in A. mexicana subsp. xolocotziana. Furthermore, a different composition was found in their respective hexane extracts. These chemical composition dissimilarities between the two taxa support their recognition as distinct subspecies.     

Rubrimonas shengliensis sp. nov. and Polymorphum gilvum gen. nov., sp. nov., novel members of Alphaproteobacteria from crude oil contaminated saline soil

Four bacterial strains were isolated from a crude oil contaminated saline soil in Shengli Oilfield, China. Strains SL014B-28A2^T and SL014B-80A1 were most closely related to Rubrimonas cliftonensis OCh 317^T, while strains SL003B-26A1^T and SL003B-26A2 were most closely related to but readily different from the species in the Pannonibacter-Labrenzia-Roseibium-Stappia cluster. The major fatty acids were C_18:1ω7c, C_16:0, C_18:0 and 11-Methyl C_18:1ω7c, and C_18:1ω7c, 11-Methyl C_18:1ω7c and C_18:0, respectively, for these two groups of isolates. Q-10 was the predominant ubiquinone. The G + C contents of genomic DNA of the four isolates were 67.9, 69.7, 65.6 and 65.6 mol%. Based on the polyphasic taxonomic characteristics, strains SL014B-28A2^T and SL014B-80A1 represented a novel species of the genus Rubrimonas, for which the name Rubrimonas shengliensis sp. nov. is proposed, with strain SL014B-28A2^T (=LMG 26072^T = CGMCC 1.9170^T) as the type strain. Isolates SL003B-26A1^T and SL003B-26A2 represented a novel genus and species of the family Rhodobacteraceae, for which the name Polymorphum gilvum gen. nov., sp. nov. is proposed, with strain SL003B-26A1^T (=LMG 25793^T = CGMCC 1.9160^T) as the type strain.     

Dual Roles of Cadaverine-Producing Pseudomonas sp. on Microcystis spp. in Hyper-Eutrophic Water

A bacterium isolated from Lake Taihu was identified as Pseudomonas sp. A3CT, which performed different effects on Microcystis spp. Growth of Microcystis flos-aquae and Microcystis aeruginosa was assessed in co-culture with A3CT to determine the stimulatory or inhibitory effects on these toxic, bloom-forming Microcystis strains. Results demonstrated that the impacts of A3CT were species specific. A3CT promoted the growth of M. aeruginosa but inhibited growth of M. flos-aquae. To investigate the cause of this phenomenon, the chemical composition of A3CT exudates and the impact of exposure to A3CT exudates on the two Microcystis species were determined. Results suggested that the observed differential growth responses of the two microalgae to A3CT exposure might be related to two components in A3CT exudates NH₄ ⁺ and cadaverine. Growth stimulation of M. aeruginosa by A3CT was significantly related to NH₄ ⁺ concentration. Cadaverine possibly acted as a growth inhibitor of M. flos-aquae. The different effects of cadaverine on growth of the two Microcystis strains suggested that A3CT might play a role in intrageneric succession patterns observed during Microcystis blooms in Lake Taihu.     

Description of the new phototrophic dinoflagellate Alexandrium pohangense sp. nov. from Korean coastal waters

The planktonic phototrophic dinoflagellate Alexandrium pohangense sp. nov. isolated from the coastal waters off Korea is described from living and fixed cells by light and scanning electron microscopy (SEM). DNA sequence data were collected from the small subunit (SSU), the large subunit (LSU), internal transcribed spacer regions (ITS1 and ITS2), and 5.8S of the ribosomal DNA (rDNA). The SSU and LSU rDNA sequences of the new dinoflagellate were 4–7% and 14–17%, respectively, different from those of Alexandrium minutum, Alexandrium ostenfeldii, Alexandrium tamutum, Alexandrium margalefii, and Alexandrium pseudogonyaulax, the most closely related species. In addition, the 5.8S rDNA sequence of the new dinoflagellate was also 12% different from those of A. minutum, A. ostenfeldii, A. tamutum, and Alexandrium peruvianum. In a phylogenetic tree based on LSU rDNA sequences, A. pohangense formed a clade with A. margalefii, and this clade was clearly distinct from the clade of A. minutum, Alexandrium diversaporum, A. tamutum, Alexandrium leei, A. ostenfeldii, and Alexandirum andersoni. Moreover, in a phylogenetic tree based on SSU rDNA sequences, A. pohangense was positioned at the base of the clade containing A. leei and A. diversaporum. Morphological analysis showed that A. pohangense has a Kofoidian plate formula of Po, 4′, 6′′, 6c, 8s, 5′′′, and 2′′′′, which confirmed its assignment to the genus Alexandrium. This dinoflagellate has a wide rectangular 1′ plate, the upper left side of which is slightly bent, protruding, and touching the 2′ plate, unlike A. margalefii, which has a wide rectangular 1′ plate that does not touch the 2′ plate, or A. pseudogonyaulax and Alexandrium camurascutulum, which have a narrower elongated pentagonal 1′ plate that touches the 2′ plate. Furthermore, the 1′ plate of A. pohangense meets the 1′′ plate as a straight vertical line, whereas that of A. camurascutulum meets the 1′′ plate as an inclined line because it is lifted by the intrusion of the 1′′ plate. In addition, A. pohangense had a relatively small ventral pore whose majority was located on the 4′ plate, unlike A. margalefii or A. pseudogonyaulax, which have a relatively large ventral pore whose majority is located on the 1′ plate. Furthermore, A. pohangense had pores of two different sizes on the cell surface, unlike A. margalefii and A. pseudogonyaulax, which have similar pores of only one size. On the basis of morphological and phylogenetic criteria, it is proposed that this is a new species of the genus Alexandrium.     

Aspergillus bertholletius sp. nov. from Brazil Nuts

During a study on the mycobiota of brazil nuts (Bertholletia excelsa) in Brazil, a new Aspergillus species, A. bertholletius, was found, and is described here. A polyphasic approach was applied using morphological characters, extrolite data as well as partial β-tubulin, calmodulin and ITS sequences to characterize this taxon. A. bertholletius is represented by nineteen isolates from samples of brazil nuts at various stages of production and soil close to Bertholletia excelsa trees. The following extrolites were produced by this species: aflavinin, cyclopiazonic acid, kojic acid, tenuazonic acid and ustilaginoidin C. Phylogenetic analysis using partial β-tubulin and camodulin gene sequences showed that A. bertholletius represents a new phylogenetic clade in Aspergillus section Flavi. The type strain of A. bertholletius is CCT 7615 ( = ITAL 270/06 = IBT 29228).     

Analysis of the interaction of Aeromonas caviae, A. hydrophila and A. sobria with mucins

Aeromonas species are known to be involved in human gastrointestinal diseases. These organisms colonize the gastrointestinal tract. Aeromonas hydrophila, A. caviae, and A. sobria have been demonstrated microscopically to adhere to animal cell lines that express mucous receptors, but quantitative studies of adherence to mucosal components such as mucin have not been published to date. Purified bovine submaxillary gland, hog gastric mucin, and fish skin mucin were used as a model to study mucin-binding activity among A. caviae, A. hydrophila, and A. sobria strains. Our findings revealed that binding of radiolabeled and enzyme-conjugated mucins to Aeromonas cells varied depending on the labeling procedure. The highest binding was observed when the three mucin preparations were labeled with horseradish peroxidase. Binding of the various horseradish peroxidase-labeled mucins by A. caviae, A. hydrophila, and A. sobria cells is a common property among Aeromonas species isolated from human infections, diseased fish, and from environmental sources. The proportion of Aeromonas strains which bind the various horseradish peroxidase-labeled mucins was significantly higher for A. hydrophila than for A. caviae and A. sobria. Bacterial cell-surface extracts containing active mucin-binding components recognized the horseradish peroxidase-labeled mucins. The molecular masses of the mucin-binding proteins were estimated by SDS-PAGE and Western blot as follows: A. caviae strain A4812 (95 and 44 kDa); A. hydrophila strain 48748 (97, 45, 33 and 22 kDa); and A. sobria strain 48739 (95 and 43 kDa). Mucin interaction with Aeromonas cells was also studied in terms of growth in mucin-rich media. The culture conditions greatly influence the expression of A. hydrophila mucin-binding activity.     

The genus Aleurodiscus s.l. (Stereaceae, Russulales) in the Patagonian Andes

The genus Aleurodiscus s.l. in Patagonia is revised and ten species are reported, commented on, and illustrated. Aleurodiscus bernicchiae, A. corticola, A. hallenbergii, A. quilae, and A. stratosus, are proposed as new. Aleurodiscus parmuliformis is recorded for the first time out of New Zealand. A key to the species of Aleurodiscus from Patagonia is included.     

Taxonomic revision of European Apium L. s.l.: Helosciadium W.D.J.Koch restored

European representatives of Apium sensu lato (Apiaceae), and Apium prostratum and Naufraga balearica, were studied with morphological, fruit anatomical, and palynological methods. Morphometric data were compared with phylogenetic results from previous molecular studies. This confirms that most of the European Apium species belong to a separate group corresponding to the previously named genus Helosciadium. All these species had previously been formally named as Helosciadium species, except for the new combination Helosciadium bermejoi, which is formally described here. Molecular studies place Apium prostratum and Naufraga balearica close to Apium graveolens, the type species of Apium. Our morphometric results show similarities of Naufraga with H. bermejoi, but fruit anatomy distinguishes it both from Helosciadium and from A. graveolens/prostratum. The placement of Cyclospermum leptophyllum in a separate genus is confirmed. Diagnostic keys to the genera and Helosciadium species, and an annotated checklist are given.     

A new putrescine bisamide phenolic glycoside from the seeds of Lallemantia iberica (M. Bieb.) Fisch. & C. A. Mey

A new putrescine bisamide phenolic glycoside, N-(trans-feruloyl)-N′-(para-hydroxybenzoyl) putrescine bisamide-4′-O-α-l-rhamnopyranoside (1), designated as lallenmantoside (1), together with one known phenolic glycoside, cucurbitoside d (2), were isolated from the seeds of Lallemantia iberica (M. Bieb.) Fisch. & C. A. Mey. Their structures were elucidated by spectroscopic analysis, and by comparison with literature data. This is the first report of phenolic glycosides from genus Lallemantia.     

Parablastocatena tetracerae gen. et sp. nov. and Corynesporella licualae sp. nov. from Hainan, China

Parablastocatena tetracerae gen. et sp. nov. and Corynesporella licualae sp. nov., collected on dead branches of Tetracera asiatica and Licuala fordiana, respectively, in tropical forests of China, are described and illustrated. Parablastocatena tetracerae is the type species for a new monotypic genus in possessing macronematous conidiophores forming distinct synnemata with holoblastic conidiogenesis and euseptate, short-chained conidia ending in a paler brown rostrum, whereas C. licualae is distinguished from described species by the smaller conidia with long appendages. A key to currently accepted species of Corynesporella is provided.     

Differential hygienic behavior of Apis cerana F. and Apis mellifera L. to Sacbrood virus infection

Beekeeping with Apis cerana of Korean apiculture is facing with serious colony collapse caused by invasive Sacbrood virus (SBV) disease. This fatal brood disease was the main reason of more than 90% colony lost in Korea leading almost the extinct crisis. Sacbrood virus can infect either larvae or adult honeybees, with a higher sensibility of larvae to the infection. Since SBV has spread to all over the country, efforts have been made to treat and prevent this devastating disease although no effective results have so far been obtained. Several studies have demonstrated that Apis mellifera bee colonies that express an efficient hygienic behavior exhibit a higher resistance to the brood disease. In this study we demonstrated that the differences of hygienic behavior between A. cerana and A. mellifera. A. cerana more efficiently removed the pin-killed brood than A. mellifera. On the other hand, A. mellifera more efficiently removed SBV-infected larvae and SBV-dead brood than A. cerana. However, it remains unclear whether the advantage of hygienic bee could have efficacy against Sacbrood disease on A. cerana colonies.     

Molecular phylogeny and new taxa in the Archaeosporales (Glomeromycota): Ambispora fennica gen. sp. nov., Ambisporaceae fam. nov., and emendation of Archaeospora and Archaeosporaceae

The AM fungal family Archaeosporaceae and the genus Archaeospora are rendered paraphyletic by the relationship with the Geosiphonaceae. This problem led to a more detailed study of the Archaeosporales. Members of the Archaeosporaceae were described as forming both glomoid and acaulosporoid spores, or solely acaulosporoid spores. However, we found that Glomus callosum fell into the same phylogenetic clade as A. leptoticha and A. gerdemannii, but exclusively formed glomoid spores. To resolve these inconsistencies, a genus, Ambispora gen. nov., typified by Ambispora fennica sp. nov., is erected based on morphological evidence and SSU and ITS region rDNA data. Ambispora contains three species known to produce both acaulosporoid and glomoid spores: A. fennica, A. leptoticha comb. nov. (basionym G. leptotichum), and A. gerdemannii comb. nov. (basionym G. gerdemannii). Another species, A. callosa comb. nov. (basionym G. callosum), is known only from glomoid spores. Ambispora is placed in a new family, the Ambisporaceae fam. nov. The Archaeosporaceae is maintained with the type species, Archaeospora trappei (basionym Acaulospora trappei), along with Intraspora schenckii (basionym Entrophospora schenckii). Acaulospora nicolsonii, known only from acaulosporoid spores, is discussed and is considered likely to belong in the Ambisporaceae, but is retained within its present genus because of inadequate morphological information and a lack of molecular data.     

Antennal sensilla in adult Agathiphaga vitiensis dumbl. and A. queenslandensis dumbl. (Lepidoptera : Agathiphagidae)

The antennae of both sexes of Agathiphaga vitiensis and the antenna of the female of A. queenslandensis (Lepidoptera : Agathiphagidae) were studied with a scanning electron microscope. In both sexes, 10 types of sensilla were observed on the antennae of A. vitiensis: Böhm's bristles, chaetica (types I and II), squamiformia, trichodea, biforked basiconica, short basiconica and coeloconica (types I, II and III). The female of A. queenslandensis possesses the same types of sensilla, except that the biforked basiconica are flattened. Biforked sensillum basiconicum is being reported in Lepidoptera for the first time. Their presence supports Kristensen's hypothesis on the phylogeny of Lepidoptera (1984). The female of A. vitiensis possesses more sensilla than that of A. queenslandensis. The male of A. vitiensis has a greater number of s. trichodea than the female, but the biforked sensilla basiconica are more numerous on the female antenna than on the male one.     

Variability of Actinocyclus kuetzingii (A.S.) Sim. in the Littorina sediments of the Polish Baltic Coast

The variability of Actinocyclus kuetzingii (A.S.) Sim. in the Littorina deposits of the Vistula Lagoon and Lake Druzno were studied. During the time of the Littorina Lake Druzno was a part of the Vistula Lagoon. In the different parts of the Lagoon both the abundance of A. kuetzingii and the diameter of its valves change in a manner associated with phases of the Littorina transgression. Coscinodiscus (Actinocyclus) kuetzingii fo. subsalsa Brockmann is probably an ecotype of A. kuetzingii (A.S.) Sim.     

Comparative Morphology of Atalodera spp. and Thecavermiculatus spp. (Heteroderidae) with Scanning Electron Microscopy

SEM examination of second-stage juveniles (J2) and adults of Atalodera ucri, A. lonicerae (syn. Sherodera lonicerae), Thecavermiculatus sp. (undescribed new species), T. andinus, and T. crassicrustatus revealed new characters. A primitive en face pattern with six separate lips occurs in J2 of Thecavermiculatus spp. examined and in about half the polymorphic A. lonicerae. A derived en face pattern with fused adjacent submedial lips occurs in the other half of A. lonicerae and all A. ucri. Posteriorly, the J2 head of all species is annulated. The primitive en face pattern also occurs in males of A. lonicerae and Thecavermiculatus spp., and posteriorly the head of these species consists of plates. Fewer plates occur rarely in males of A. ucri. Males of A. ucri have a derived en face pattern where lips are fused and the head is annulated. Fusion of lips occurs rarely in males of A. lonicerae. Females of all species have similar derived en face patterns. En face patterns of J2 and males o f Atalodera and Thecavermiculatus may aid in species identification and to elucidate intergeneric relationships, but en face characters shared by the two genera are primitive and are not useful for demonstrating monophyly. Perineal region of females indicates the closeness of the vulval-anal distance, as a derived character, which is shared by Atalodera and most Thecavermiculatus spp. suggesting possible monophyly. T. andinus, while having a similar en face pattern to J2 of other Thecavermiculatus species, lacks the derived character of the perineal region. Phasmid openings were not observed in adults of any of the species examined.