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.     

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.     

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.     

A Novel Hyaluronidase Produced by Bacillus sp. A50

Hyaluronidases are a family of enzymes that degrade hyaluronic acid (hyaluronan, HA) and widely used in many fields. A hyaluronidase producing bacteria strain was screened from the air. 16S ribosomal DNA (16S rDNA) analysis indicated that the strain belonged to the genus Bacillus, and the strain was named as Bacillus sp. A50. This is the first report of a hyaluronidase from Bacillus, which yields unsaturated oligosaccharides as product like other microbial hyaluronate lyases. Under optimized conditions, the yield of hyaluronidase from Bacillus sp. A50 could reach up to 1.5×10⁴ U/mL, suggesting that strain A50 is a good producer of hyaluronidase. The hyaluronidase (HAase-B) was isolated and purified from the bacterial culture, with a specific activity of 1.02×10⁶ U/mg protein and a yield of 25.38%. The optimal temperature and pH of HAase-B were 44°C and pH 6.5, respectively. It was stable at pH 5–6 and at a temperature lower than 45°C. The enzymatic activity could be enhanced by Ca²⁺, Mg²⁺, or Ni²⁺, and inhibited by Zn²⁺, Cu²⁺, EDTA, ethylene glycol tetraacetic acid (EGTA), deferoxamine mesylate salt (DFO), triton X-100, Tween 80, or SDS at different levels. Kinetic measurements of HAase-B towards HA gave a Michaelis constant (K _m) of 0.02 mg/mL, and a maximum velocity (V _max) of 0.27 A ₂₃₂/min. HAase-B also showed activity towards chondroitin sulfate A (CSA) with the kinetic parameters, K _m and V _max, 12.30 mg/mL and 0.20 A ₂₃₂/min respectively. Meanwhile, according to the sequences of genomic DNA and HAase-B’s part peptides, a 3,324-bp gene encoding HAase-B was obtained.     

Aspergillus osmophilus sp. nov., and a new teleomorph for A. proliferans

A new species of Aspergillus and a new teleomorph for A. proliferans, both isolated from cereals in Iran, are described using morphological and molecular data. A combined sequence dataset of the ITS region, partial β-tubulin and partial calmodulin genes resolved the relationships of members of section Aspergillus largely in concordance with morphological traits of ascospores. Aspergillus osmophilus sp. nov. is differentiated from the closest species, A. xerophilus by possessing larger ascospores, conidia and associated fruiting bodies. Both species are strongly xerophilic and possess ascospores with lobate-reticulate convex surfaces. The newly discovered teleomorph for A. proliferans is characterized by delicately roughened ascospores with a shallow or distinct furrow and finely roughened to irregular equatorial crests.     

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.     

Presence of OXA-Type Enzymes in Achromobacter insuavis and A. dolens

The accurate species identification of Achromobacter isolates is difficult and the clinical isolates of this genus are mostly referred as A. xylosoxidans. Here, we report new OXA variants in 2 isolates identified as A. insuavis (A114, A79) and 1 isolate identified as A. dolens (A336). These results suggest that different bla _OXA genes are ubiquitous in the different species of Achromobacter spp. The role of the other species of Achromobacter in clinical samples needs to be reevaluated, and the proper identification is absolutely necessary to understand the epidemiology of this genus.     

Glossopetalon (Crossosomataceae) and a new variety of G. spinescens from the Great Basin,U.S.A.

A new variety from the eastern Great Basin of western U.S.A.,Glossopetalon spinescens var.microphyllum (Crossosomataceae), is described and illustrated, and its relationship within the species is indicated by a key. The nameGlossopetalon A. Gray is used rather than the substituteForsellesia Greene, because there is little chance of confusion with the earlier nameGlossopetalum Schreber, which was illegitimately published and remains obscure.     

Microfluorometry analysis II. A Bayesian approach.

In flow microfluorometry (FMF) analysis cells stained with a fluorescent dye that binds specifically to DNA are passed through the instrument. The number of cells in the population having a fluorescence intensity is recorded in a single channel of a multichannel pulse height analyzer. The result is a DNA fluorescence histogram for the population.A method for decomposing an FMF histogram into its G₁, S and G₂ + M components, corresponding to similarly designated phases of the cell cycle is given. This technique can also be applied to find the parameters in all of the previous approaches. The parameters are calculated by iteration which eliminates the need for non-linear optimization procedures.     

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.     

A new species of the Weevil genus Auleutes Dietz (Coleoptera, Curculionidae: Ceutorhynchinae) from the U. S. A.

A new species, Auleutes alexanderi sp. n. formerly confused with and similar to the Holarctic A. epilobii (Pk.) is described from the Middle West and Southeast United States. The new species differs from A. epilobii in the rostrum densely covered with broad white scales in males and longer, less strongly curved and matte, not striate, and less flattened in females and in the structure of the aedeagus with a broad membranous medial part along the entire length.     

Two new species ofTrichogramma [Hym.: Trichogrammatidae] from the U.S.A.

Two new species ofTrichogramma from the U.S.A. are described.T. parkeri sp. n., a morphologically distinct species, was reared from an egg ofHeliothis zea (Boddie), whileT. platneri sp. n. which is morphologically indistinguishable fromT. minutum Riley, was reared from eggs ofCydia pomonella (L.). Reproductive incompatibility ofT. platneri andT. minutum is discussed.     

Energy-dependent uptake of ochratoxin A by mitochondria.

The interaction of ochratoxin A, a mycotoxin produced by Aspergillus ochraceus, with isolated rat liver mitochondria and plasma membranes has been studied. Cell membranes bind [¹⁴C]ochratoxin A poorly and do not show saturation in the concentration range examined. The uptake of the toxin by mitochondria is saturable, with an apparent K_m at 0 °C of 30 nmol/mg of protein. Sonication or freeze-thawing reduces the extent of incorporation by 88%. Ochratoxin A uptake is energy dependent, resulting in a depletion of intramitochondrial ATP. Uncouplers such as m-chlorocarbonylcyanide phenylhydrazone or the respiratory inhibitors rotenone and antimycin A inhibit uptake 60–85%, while ATP reverses the antimycin and rotenone inhibition. Phosphate transport is sensitive to inhibition by the toxin, as measured by Ca²⁺ plus P_istimulated respiration and [³²P]P_i incorporation. In turn, phosphate inhibits nearly completely [¹⁴C]ochratoxin A uptake at 22 °C and causes a concomitant mitochondrial swelling yet is not incorporated into the matrix space. Thus, the saturable uptake of ochratoxin A is accompanied by a decrease in the energy state and inhibition of P_i transport, which results in deteriorative changes of the mitochondria, as evidenced by large-amplitude swelling.     

A comparative study of sulphur content of someAllium L. Species

Garlic has been used worldwide as a folk medicine to cure a wide range of diseases. AlthoughAllium sativum is the species commonly used, several otherAllium species have been used as a substitute for common garlic, especially in the past. This work compares the extracts from bulbs of common garlic with those of otherAllium species. TLC analysis was used to compare the sulphur compounds inA. sativum, A. subhirsutum, A. pendulinum, A. roseum, andA. triquetrum. Results have showed that the use of the above mentioned wildAllium species is not without scientific support.     

Cell surface differentiation of Chlamydomonas during gametogenesis. I. Mating and concanavalin A agglutinability

Sexual agglutination of opposite gametes and agglutination of like gametes by concanavalin A (Con A) were studied in Chlamydomonas moewusii and C. reinhardtii for the possibility that the surface sites involved in these agglutinating phenomena may be the same. Our data show that the two agglutinating phenomena appeared at different times after the beginning of gametogenesis. Sucrose and mannose block agglutination of the gametes by Con A but do not affect mating ability. Trypsin eliminates mating ability, except in (?) gametes of C. moewusii, while Con A agglutinability remains. Monovalent Con A can selectively bind to Con A-binding sites to block agglutination of gametes by multivalent Con A while mating ability is unaffected. The data indicate that the mating agglutinin and the Con A-binding sites are two different flagellar surface agglutinins that occur coincidentally during gametic differentiation of both mating types of C. moewusii and C. reinhardtii. The function of the Con A-binding site on Chlamydomonas gametes is not known.     

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.     

A melissopalynological study of 54 Louisiana (U.S.A.) honeys

Pollen analyses were made of 54 commercial Louisiana (U.S.A.) honeys collected during 1967–1968. Fifty-eight different pollen types were identified. A direct correlation was assumed between a plant's pollen quantity and its nectar contribution. On this basis, the major Louisiana honey plants that were determined are: Fabaceae, predominantly Trifolium repens (99%), Rubus, Berchemia scandens, and Salix. Plants established as important for their nectar contributions in isolated samples are: Cephalanthus occidentalis, Sapium sebiferum, miscellaneous Compositae (Asteraceae), Glycine max, and Parthenocissus quinquefolia. Native plants such as Rubus, Berchemia scandens, and Salix contribute more to Louisiana's honey sources than do cultivated plants or introduced plants. Most samples originated from six to fifteen different plant types and thus do not appear to be of particularly diverse botanical origin. The pollen types that indicate late spring, summer, or fall honeys include: Glycine max, Cassia fasciculata, Polygonum, Lagerstroemia indica, Lythrum, Ampelopsis, Trachelospermum difforme, and others. Thirty-one samples could be considered unifloral honeys. Most Louisiana honeys were a clear color and from nectar of flowers and not honeydew.     

Cleistanthane diterpenoids from the resin of Vellozia pyrantha A.A.Conc and their chemotaxonomic significance

Vellozia pyrantha A.A.Conc. is endemic to the rupestrian fields in northern Chapada Diamantina National Park (Brazil). This species has the peculiar characteristic of only producing flowers after a massive fire event occurs in the surrounding areas. This is the first report on the assessment of the chemical composition of Vellozia pyrantha resin, where five cleistanthane diterpenoids (1–5) were identified. Their structures were proposed after careful analysis of ¹H and ¹³C NMR, DEPT 135, HMBC, HMQC, and COSY spectra as well as comparison with the literature. Cleistanthane diterpenoids show great the potential to be used as chemosystematics biomarkers for the genus Vellozia and, therefore, their chemotaxonomic significance is discussed.