On antigenic O-relationships between the groupsSalmonella,Arizona, Escherichia andShigella

Summary An account is given of O-relationships between the groupsSalmonella (O-antigens 1–52),Arizona (O-antigens 1,2–1,33)Escherichia (O-antigens 1–142 and OX 1–OX 13) andShigella (O-antigens A1–A10, B 1a–B6, C1–C15 and D). Through cross agglutination studies and absorption tests the instances of O identity in antigens of the different groups were determined. This work has been performed during a stay in the Enteric Bacteriology Unit of the Communicable Disease Centre, Atlanta (Ga.), U.S.A. It was sponsored by the Fulbright Organisation and the Department of Social Affairs and Public Health, The Netherlands.     

Antigenic relationship betweenHaemophilus influenzae andHaemophilus isolated from porcine pleuropneumonia

Comparative double immunodiffusion techniques were used to study capsular and O antigenic relationships betweenHaemophilus influenzae types a-fandH. pleuropneumoniae types 1–5 and a strain (202) closely related toH. pleuropneumoniae. Culture fluids or culture supernatants were used as antigens and rabbit antisera were produced against cell suspensions of the strains tested. A reaction of identity was obtained between the capsular precipitate ofH. influenzae c and a precipitate formed by strain 202, when developed with anti-H. influenzae c serum or the serum produced against strain 202. Mutual cross-absorption of capsular antibodies was also demonstrable. No other capsular or O antigenic cross-reactivity was demonstrable between the strains tested.     

All-electron scalar relativistic calculation of water molecule adsorption onto small gold clusters

An all-electron scalar relativistic calculation was performed on Au _n H₂O (n = 1–13) clusters using density functional theory (DFT) with the generalized gradient approximation at PW91 level. The calculation results reveal that, after adsorption, the small gold cluster would like to bond with oxygen and the H₂O molecule prefers to occupy the single fold coordination site. Reflecting the strong scalar relativistic effect, Au _n geometries are distorted slightly but still maintain a planar structure. The Au–Au bond is strengthened and the H–O bond is weakened, as manifested by the shortening of the Au–Au bond-length and the lengthening of the H–O bond-length. The H–O–H bond angle becomes slightly larger. The enhancement of reactivity of the H₂O molecule is obvious. The Au–O bond-lengths, adsorption energies, VIPs, HLGs, HOMO (LUMO) energy levels, charge transfers and the highest vibrational frequencies of the Au–O mode for Au _n H₂O clusters exhibit an obvious odd-even oscillation. The most favorable adsorption between small gold clusters and the H₂O molecule takes place when the H₂O molecule is adsorbed onto an even-numbered Au _n cluster and becomes an Au _n H₂O cluster with an even number of valence electrons. The odd–even alteration of magnetic moments is observed in Au _n H₂O clusters and may serve as material with a tunable code capacity of “0” and “1” by adsorbing a H₂O molecule onto an odd or even-numbered small gold cluster.     

Pathogen specific carbohydrate antigen microarrays: a chip for detection of Salmonella O-antigen specific antibodies

A Salmonella O-antigen microarray was developed by covalent coupling of oligosaccharide antigens specific for serogroups Salmonella enterica sv. Paratyphi (group A), Typhimurium (group B) and Enteritidis (group D). Antibodies were correctly detected in sera from patients with culture verified salmonellosis. High serogroup-specificity was seen with the disaccharide antigens. With the larger antigens, containing the backbone sequence Manα1–2Rhaα1–2Gal (MRG), common backbone-specific antibodies (O-antigen 12) were also detected. This is “proof of principle” that pathogen-specific carbohydrate antigen microarrays constitute a novel technology for rapid and specific serological diagnosis in either individual patients or larger sero-epidemiological and vaccine studies.     

Islet cell antigens and autoantigen(s): Monoclonal antibodies and further characterization

A novel series of murine monoclonal antibodies to islet cells (1–45, 1–51, 1–52 and 1–39) have been generated using human insulinoma homogenate as the immunogen in order to characterize pathogenetically relevant islet cell autoantigen(s). Differentiation antigens recognized by these islet cell monoclonal antibodies displayed varied cytological distribution (pan-islet or peripheral mantle only). Monoclonal antibody 1–45 reacted with all endocrine subsets of the pancreatic islet, similar to the reactivity of islet cell autoantibody positive sera from type I diabetes subjects. Preexposure to pH2 abolished the immunoreactivity of the autoantigen; 1–45 antigen was also sensitive to low pH. Preexposure to 100° C for 1 h did not significantly alter the immunoreactivity of islet antigens recognized by ICAb positive patient sera and monoclonal antibody 1–39, thus demonstrating the extraordinary heat stability of the corresponding epitopes; those recognized by 1–45 were less heat stable. Islet cells were found to share 1–45 differentiation antigen(s)/epitope(s) with other neuroendocrine cells,viz. amerior pituitary, adrenal medulla and gut endocrine cells.     

Microsolvation effect and hydrogen-bonding pattern of taurine-water TA-(H2O)n (n = 1-3) complexes

The microsolvation of taurine (TA) with one, two or three water molecules was investigated by a density functional theory (DFT) approach. Quantum theory of atoms in molecules (QTAIM) analyses were employed to elucidate the hydrogen bond (H-bond) interaction characteristics in TA-(H₂O)_n (n = 1–3) complexes. The results showed that the intramolecular H-bond formed between the hydroxyl and the N atom of TA are retained in most TA-(H₂O)_n (n = 1–3) complexes, and are strengthened via cooperative effects among multiple H-bonds from n = 1–3. A trend of proton transformation exists from the hydroxyl to the N atom, which finally results in the cleavage of the origin intramolecular H-bond and the formation of a new intramolecular H-bond between the amino and the O atom of TA. Therefore, the most stable TA-(H₂O)₃ complex becomes a zwitterionic complex rather than a neutral type. A many-body interaction analysis showed that the major contributors to the binding energies for complexes are the two-body energies, while three-body energies and relaxation energies make significant contributions to the binding energies for some complexes, whereas the four-body energies are too small to be significant.     

Detection of immunomagnetically captured Escherichia coli O157:H7 by antibody-conjugated alkaline phosphatase

A rapid and sensitive detection process for Escherichia coli O157:H7 was developed using alkaline phosphatase (APase)-labeled anti-E. coli O157 antibodies to tag the targeted bacteria. Immunomagnetic beads or antibody-labeled streptavidin-coated magnetic beads were then used to capture the APase-tagged E. coli. Immunomagnetically captured bacteria were washed and distributed into microplates or optical cuvettes. The enzyme-catalyzed hydrolysis of p-nitro-phenol phosphate in alkaline solutions was then followed. Less than 1000 cfu/ml of E. coli O157:H7 could be detected. This approach was applied to detect the bacteria artificially spiked in beef hamburgers. Less than 1 cfu/g of E. coli O157:H7 produced a significant response after cultural enrichment for 4–6 h at 37°C. Journal of Industrial Microbiology & Biotechnology (2001) 26, 345–349. Received 30 August 2000/ Accepted in revised form 17 April 2001     

Further reports of Acanthocolpidae Lühe, 1906 (Digenea) from fishes off New Caledonia, with descriptions of two new species

Ophiotaenia bungari n. sp. (Cestoda: Proteocephalidea) is described from the intestine of the banded krait Bungarus fasciatus (Schneider) (Ophidia: Elapidae) in Vietnam. The new species differs from all but three Ophiotaenia species parasitic in Asian reptiles in the possession of a glandular apical organ. It differs from O. andersoni Jensen, Schmidt & Kuntz, 1983 in the position of the vagina in relation to the cirrus-sac (anterior and posterior in O. bungari versus anterior only in the latter species), in the cirrus-sac/proglottis width ratio (29–38 versus 50%) and by having more testes (100–150 versus 42–116 in O. andersoni); from O. chattoraji Srivastava, 1980 in the number of uterine diverticula (50–65 versus 10–26) and in the cirrus-sac/proglottis width ratio (29–38 versus 22%); and from O. rhabdophidis (Burt, 1937) by having more uterine diverticula (50–65 versus 30–45), by the cirrus-sac/proglottis width ratio (29–38 versus 20–25%) and by the width of the scolex (360–420 versus 130–187 μm). The taxonomic importance of the relative size of the ovary (i.e. the ratio of the ovarian size in relation to that of the entire proglottis), a character previously not used in the systematics of proteocephalidean cestodes, is discussed. Comparison of measurements of all of the nominal species of Ophiotaenia La Rue, 1911 and Proteocephalus Weinland, 1858 (c.135 species) has shown that the ovary of species parasitic in snakes in the Americas, Africa, Asia and Australia is not only considerably smaller than that of congeneric species from European hosts, but also smaller than in all species of Proteocephalus parasitic in teleost fishes throughout the world.     

Theoretical study of the hydroxylation of phenolates by the Cu2O2(N,N′-dimethylethylenediamine)2 2+ complex

Tyrosinase catalyzes the ortho hydroxylation of monophenols and the subsequent oxidation of the diphenolic products to the resulting quinones. In efforts to create biomimetic copper complexes that can oxidize C–H bonds, Stack and coworkers recently reported a synthetic μ-η²:η²-peroxodicopper(II)(DBED)₂ complex (DBED is N,N′-di-tert-butylethylenediamine), which rapidly hydroxylates phenolates. A reactive intermediate consistent with a bis-μ-oxo-dicopper(III)-phenolate complex, with the O–O bond fully cleaved, is observed experimentally. Overall, the evidence for sequential O–O bond cleavage and C–O bond formation in this synthetic complex suggests an alternative mechanism to the concerted or late-stage O–O bond scission generally accepted for the phenol hydroxylation reaction performed by tyrosinase. In this work, the reaction mechanism of this peroxodicopper(II) complex was studied with hybrid density functional methods by replacing DBED in the μ-η²:η²-peroxodicopper(II)(DBED)₂ complex by N,N′-dimethylethylenediamine ligands to reduce the computational costs. The reaction mechanism obtained is compared with the existing proposals for the catalytic ortho hydroxylation of monophenol and the subsequent oxidation of the diphenolic product to the resulting quinone with the aim of gaining some understanding about the copper-promoted oxidation processes mediated by 2:1 Cu(I)O₂-derived species. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Bacterial siderophores: the solution stoichiometry and coordination of the Fe(III) complexes of pyochelin and related compounds

Pyochelin, its analog 3′′-nor-NH-pyochelin, and the related methyl hydroxamate, 2-(2′-hydroxyphenyl)-4,5-dihydrothiazol-4-carboxylic acid methoxymethyl amide, have been prepared together with their Fe(III) complexes. The solution stoichiometry and the coordination of the three Fe(III) complexes in methanol or buffered (pH∼2) 50:50 (v/v) methanol–water mixtures were determined using various spectroscopic methods: UV–vis absorption, X-ray absorption, extended X-ray absorption fine structure and electron paramagnetic resonance. All three systems showed both a 1:1 and 2:1 ligand–Fe(III) stoichiometry, but presented different coordination properties. Conditional formation constants (pH∼2) were determined for both the 1:1 and 2:1 complexes in all three systems. Computation of the coordination-conformational energies by semiempirical methods indicated that the coordination in the case of the 2:1 complexes of pyochelin–Fe(III) and 3′′-nor-NH-pyochelin–Fe(III) was asymmetrical, with one molecule of pyochelin (or 3′′-nor-NH-pyochelin) tetradentately coordinated (O1, N1, N2 and O3) to the Fe(III), and the second molecule bound bidentately (O1, N1 or N2, O3), to complete the octahedral geometry. In contrast, two molecules of the methyl hydroxamate each provided a set of tridentate ligand atoms in the formation of the 2:1 ligand–Fe(III) complex. These results are consistent with the role of pyochelin in the uptake of iron by the FptA receptor in the outer membrane of Pseudomonas aeruginosa and in several gram-negative bacteria.     

Local and systemic antibody response in infants after oral administration of inactivated enteropathogenicE. coli serotype O111 and O55

In ten infants divided into two groups (up to one month of age and at 2–7 months of age) the dynamics and formation of different antibody isotypes produced locally in the intenstine and in serum after orally administered inactivated enteropathogenicE. coli strains O111 and O55 was followed during 30 d after the first and booster dose by using an indirect immunofluorescence method. Infants up to one month of age produced antibodies of IgM isotype in stool together with the IgA isotype after the first and booster dose of the vaccine against both antigens. Serum IgG antibody increased after 2 d following the first and second dose of antigens and remained higher during 5 d. The infants aged 2–7 months expressed predominantly the IgA isotype response in stool after the first and booster dose of antigens. The serum immunoglobulin levels did not change after oral antigen administration.     

BAC/YAC contigs from the H2-M region of mouse Chr 17 define gene order as Znf173-Tctex5-Mog-D17Tu42-M3-M2

 A yeast artificial chromosome (YAC) contig from the C57BL/6 (H2 ^b ) mouse was created from the major histocompatibility complex (Mhc, H2 in mouse) class Ib subregion, H2-M. It spans approximately 1.2 megabase (Mb) pairs and unites the previous >1.5-Mb YAC contigs (Jones et al. 1995) into a single contig, which includes 21 Mhc class I genes distal to H2-T1. A bacterial artificial chromosome (BAC) contig from the 129 (H2 ^bc ) mouse, spanning approximately 600 kilobases, was also built from Znf173 (Afp, a gene for acid finger protein), through Tctex5 (t-complex testis expressed-5) and Mog (myelin oligodendrocyte glycoprotein), to H2-M2. Twenty-four sequence-tagged site (STS) markers were newly developed, and 35 markers were mapped in the YAC/BAC contigs, which define the marker order as Cen –Znf173–Tctex5 – Mog–D17Tu42–D17Mit232–H2-M3–D17Leh525–H2-M2– Tel. The gene order of Znf173 – Tctex5 – Mog – D17Tu42 is conserved between mouse and human, showing that the middle H2-M region corresponds to the subregion of the human Mhc surrounding HLA-A. Received: 25 July 1997 / Revised: 10 September 1997     

Organic matter stabilization in soil microaggregates: implications from spatial heterogeneity of organic carbon contents and carbon forms

This study investigates the spatial distribution of organic carbon (C) in free stable microaggregates (20–250 μm; not encapsulated within macroaggregates) from one Inceptisol and two Oxisols in relation to current theories of the mechanisms of their formation. Two-dimensional micro- and nano-scale observations using synchrotron-based Fourier-transform infrared (FTIR) and near-edge X-ray absorption fine structure (NEXAFS) spectroscopy yielded maps of the distribution of C amounts and chemical forms. Carbon deposits were unevenly distributed within microaggregates and did not show any discernable gradients between interior and exterior of aggregates. Rather, C deposits appeared to be patchy within the microaggregates. In contrast to the random location of C, there were micron-scale patterns in the spatial distribution of aliphatic C–H (2922 cm⁻¹), aromatic C=C and N–H (1589 cm⁻¹) and polysaccharide C–O (1035 cm⁻¹). Aliphatic C forms and the ratio of aliphatic C/aromatic C were positively correlated (r ² of 0.66–0.75 and 0.27–0.59, respectively) to the amount of O–H on kaolinite surfaces (3695 cm⁻¹), pointing at a strong role for organo-mineral interactions in C stabilization within microaggregates and at a possible role for molecules containing aliphatic C-H groups in such interactions. This empirical relationship was supported by nanometer-scale observations using NEXAFS which showed that the organic matter in coatings on mineral surfaces had more aliphatic and carboxylic C with spectral characteristics resembling microbial metabolites than the organic matter of the entire microaggregate. Our observations thus support models of C stabilization in which the initially dominant process is adsorption of organics on mineral surfaces rather than occlusion of organic debris by adhering clay particles.     

Alkalilactibacillus ikkensis, gen. nov., sp. nov., a novel enzyme-producing bacterium from a cold and alkaline environment in Greenland

Three novel Gram-positive, endospore-forming bacteria were isolated from a cold and alkaline environment. Phylogenetic analysis showed that the strains were almost identical, and that they were related to Natronobacillus azotifigens 24KS-1^T (95.8% identity), Paraliobacillus quinghaiensis YIM-C158^T (95.1%), Paraliobacillus ryukyuensis O15-7^T (94.5%), and Halolactibacillus miurensis M23-1^T (93.9%). The isolates produced amylase, α-galactosidase, β-galactosidase, and β-glucuronidase, and showed optimal growth at pH 10, at 20°C, and at 2–8% (w/v) NaCl. Major fatty acids were C_14:0 (10.6–11.6%), anteiso-C_15:0 (25.7–32.7%), C_16:1 ω11c (12.2–16.0%), and C_16:0 (14.0–20.4%). The major polar lipids were diphosphatidylglycerol and phosphatidylglycerol, and meso-diaminopimelic acid was found in the cell-wall peptidoglycan. The G+C content was 38.4%. DNA–DNA hybridization between strain GCM68^T and H. miurensis M23-1^T was 32.4%, while hybridization to N. azotifigens 24KS-1^T, Amphibacillus tropicus Z-7792^T, and Paraliobacillus ryukyuensis O15-7^T was below 30%. The phylogenetic analysis and G+C content place strain GCM68^T in relation to species belonging to Bacillus rRNA group 1, but phylogenetic and physiologic data combined with chemotaxonomic analyses support our proposal for a new genus, Alkalilactibacillus, gen. nov., with the novel species Alkalilactibacillus ikkensis, sp. nov. (type strain is GCM68^T = DSM 19937 = LMG 24405).     

Climate signals in the ring widths and stable carbon, hydrogen and oxygen isotopic composition of Larix decidua growing at the forest limit in the southeastern European Alps

The southeastern border of the European Alps is not well resourced with high-resolution climate proxies and experiences a distinct climatic regime from the northern and western Alpine zones. Here, we present new high-resolution climatic proxies (AD 1907–2006) from ring widths and stable carbon (δ¹³C), non-exchangeable hydrogen (δ²H) and oxygen (δ¹⁸O) isotope ratios of cellulose extracted from Larix decidua tree rings, growing at the forest limit in the southeastern European Alps (Slovenia). δ¹³C, δ²H and δ¹⁸O are strongly (p < 0.001) and positively correlated with each other. June temperature has the strongest control on tree ring width (TRW), while later summer conditions (July–August) influence the stable isotope composition. All four proxies are strongly correlated (r > 0.4; p < 0.001) with summer temperature and also sunshine hours, while precipitation has less impact. A combination of TRW and δ¹³C provides the greatest potential for reconstructing past temperatures (June–August) with significant (p < 0.001) correlations with gridded temperatures extending across a very large part of southern and western Europe west of the Carpathian Mountains. The water isotopes (oxygen and hydrogen) record conditions in the Adriatic and Mediterranean, which are the source area for the air masses that bring precipitation to this region giving strong correlations with temperatures in southern Italy and the western part of the Balkan Peninsula. Combining proxies with different spatial and temporal signals allows the strength and spatial footprint of climate signals to be enhanced. These findings open new perspectives for climate reconstruction in the southeastern European Alps and Western Balkans.     

Simultaneous detection of Salmonella spp and Escherichia coli O157:H7 by multiplex PCR

Contamination of foods with pathogens such as Escherichia coli O157:H7 and Salmonella is a major concern worldwide and rapid, sensitive, and reliable methods are needed for detection of these organisms. Since these pathogens can contaminate similar foods and other types of samples, a multiplex polymerase chain reduction (PCR) was designed to allow simultaneous detection of both E. coli O157:H7 and Salmonella spp directly from enrichment cultures. Samples of apple cider, beef carcass wash water, ground beef, and bovine feces were inoculated with both E. coli O157:H7 and S. typhimurium at various bacterial levels. Following enrichment culturing for 20–24 h at 37°C in modified EC broth or buffered peptone water both containing novobiocin, the samples were subjected to a DNA extraction technique or to immunomagnetic separation then tested by the multiplex PCR assay. Four pairs of primers were employed in the PCR: primers for amplification of E. coli O157:H7 eaeA, stx _1/2 and plasmid sequences and for amplification of a portion of the Salmonella invA gene. Four fragments of the expected sizes were amplified in a single reaction and visualized following agarose gel electrophoresis in all the samples inoculated with ≤ 1 CFU g⁻¹ or ml⁻¹. Results can be obtained in approximately 30 h. The multiplex PCR is a potentially powerful technique for rapid and sensitive co-detection of both pathogens in foods and other types of samples. Received 28 December 1997/ Accepted in revised form 19 March 1998     

Hemimyzon nanensis, a new balitorid fish from the chao phraya basin, thailand

A new balitorid fish,Hemimyzon nanensis, is described from the Mae Nam Nan River basin, a tributary of the Chao Phraya River, in northern Thailand. This species is distinguishable from its congeners by the following unique combination of characters: pectoral fin with 9–11 simple and 10–12 branched, rays; pelvic fin with 3–4 (usually 3) simple and 8–9 (usually 9) branched rays; 59–63 total lateral line scales; distance between pelvic fin bases 8.1–9,6% SL.H. nanensis is the first species of the genus recorded outside China and Taiwan.     

<Emphasis Type="Italic">Hirsutella proturicola</Emphasis> sp. nov. isolated from a proturan, <Emphasis Type="Italic">Baculentulus</Emphasis> densus (Protura,Hexapoda)

A new species of Hirsutella, H. proturicola, isolated from a subterranean proturan (Baculentulus densus; Protura, Hexapoda), is described and illustrated. Hirsutella proturicola is characterized by producing monoblastic phialides of 24–51.5 × 2.5–5 μm with a slightly roughened neck, fusiform and curved conidia of 9–18 × 2.5–4 μm that have a truncate base and a papillate projection often capped with sheath-like mucilage, and pluricellular, globose to subglobose chlamydospores of 21–48 × 21–41.5 μm. This species is morphologically and phylogenetically close to H. rostrata, an acaropathogenic species, but can be distinguished from the size of the phialides and the size and shape of the conidia.     

New O and H antigens ofPlesiomonas shigelloides and their O antigenic relationships toShigella boydii

Twenty six new O (O51-O76) and 24 new H (H18-H41) antigens forPlesiomonas shigelloides serotyping scheme were defined. The O antigens 54 and 57 were closely related to those ofShigella boydii 2 and 9, respectively, in an a, b-a, c type of relationship.     

Species of criconematids (Nematoda: Criconematina) from Haryana,India

Ten known, viz. Criconema (Nothocriconemella) mutabile (Taylor, 1936) Raski & Luc, 1985, Neolobocriconema aberrans (Jairajpuri & Siddiqi, 1963) Andrássy, 1979, Macroposthonia sphaerocephala (Taylor, 1936) De Grisse & Loof, 1965, Nothocriconemoides basili (Jairajpuri, 1963) Loof & De Grisse, 1974, Hemicriconemoides cocophillus Loos, 1949, H. litchi Edward & Misra, 1963, H. mangiferae Siddiqi, 1961, Aulosphora indica (Siddiqi, 1961) Siddiqi, 1980, Paratylenchus obtusicaudatus Raski, 1975, and Tylenchulus semipenetrans Cobb, 1913, plus three new species of criconematids are recorded from Haryana state, India. Larvae of N. aberrans and N. basili are described for the first time. The main distinguishing features of the new species are as follows. M. paronostris n. sp.: R=117–128, R V=7–9, spear=55–62 μm, submedian lobes present, larvae with smooth annules; Criconemoides punicus n. sp.: R=75–85, R V=5–7, spear=73–81 μm, larvae with crenate annules: Hemicycliophora siddiqii n. sp.: R=185–198, R V=33–42, spear=57–59 μm, one incisure plus longitudinal lines on the cuticle. ac]19860702