Four newSalmonella types (S. tilburg,S. bunnk,S. overschie andS. sterrenbos) isolated in the Netherlands

Summary Four newSalmonella types isolated in the Netherlands are described.S.tilburg, 1,3,19; d∶1, w, was isolated from the lymphnodes of a swine.S.bunnik, 43; z42:-, was isolated from a reptile.S.overschie, 51; 1,v∶1,5 was isolated from a tortoise.S.sterrenbos, 6,8;d: enx, was isolated from the lymphnodes of a patient.     

Six newSalmonella types,isolated in ghana (S. volta,S. agona,S. wa,S. techimani,S. mampong andS. tafo)

Summary Six newSalmonella types isolated in Ghana are described.S. volta, 11: 4: 1,z₁₃, z₂₈ was isolated from a swine;S.agona 4,12: fgs:—,S.wa, 16: b: 1,5S.technimani, 28: c: z₆ andS.tafo, 1, 4 12, 27: z₃₅: 1,7 were isolated from cattle;S.mampong, 13,22: z₃₅: 1,6, was isolated from a lizzard.     

Three newSalmonella types (S. Bilthoven,S. wassenaar andS. bonaire

Summary Two newSalmonella types,S. bilthoven, 47ac: a:—andS. wassenaar, 50; gp—were isolated from reptiles in the Netherlands. A newSalmonella type,S. bonaire 50; z₄z₃₂:— was isolated from a cow on the Island Bonaire.     

Twelve newSalmonella types

Twelve hitherto unrecognizedSalmonella types are described. S. heerlen (11 : i : 1,6) andS. sloterdijk (1, 4, 12, 27 : z 35 : z 6) were isolated from patients in the Netherlands,S. maartensdijk (40 : g, p) was isolated from a healthy calf,S. maastricht (11 : z 41 : 1, 2(7)) from imported fishmeal,S. parera (11 : z 4, z 23 : -.) from water on the island Bonaire,S. putten (13, 23, 36 : d : 1, w),S. hoograven (50 : z 10 : z 6 : z 42),S. schalkwijk ((6), 14, (24) : i : e, n, ...),S. hilversum (30 : k : 1, 2) andS. harmelen (51 : z 4, z 23 : -.) from reptiles,S. breukelen (6, 8 : 1, z 13 : e, n, z 15) from a cuscus andS. maarssen (9, 46 : z 4, z 24 : z 39 : z 42) from a lizard.     

Plant regeneration from the protoplasts of Solanum tuberosum, S. nigrum and S. bulbocastanum

The mesophyll protoplasts were isolated from the Solanum tuberosum (S. tbr) clones of different ploidy level (4x Bzura cv., 2x H-8105, and 2x ZEL-1136) as well as from the wild species: S. bulbocastanum (S. blb, 2x) and two accessions of S. nigrum (S. ngr, 6x). Additionally, the protoplasts were isolated from the cell suspensions of Bzura cv. and H-8105 clone. The conditions of protoplast isolation as well as the media for their culturing and regeneration, were selected and optimized for the studied genotypes. For mesophyll protoplasts, the shooting calli were produced by all the cultured protoclones except that of S. bulbocastanum. The shoots excised from the protoplast-derived calli developed into whole plants in all the studied potato clones but only in one accession of S. nigrum, i.e. S. ngr var. gigantea. As for suspension-cell-derived protoplasts, only H-8105 clone produced the regenerative type of calli, though normal shoots could not be obtained. The regenerative capacity of the protoplasts isolated from leaves and cell suspensions is compared and discussed. We regret to report the death of M. Sc. Maria Borkowska after the completion of this work.     

Isolation and characterization of polymorphic microsatellite DNA markers in two shrew species,Sorex unguiculatus and S. caecutiens

We isolated six microsatellite markers from the partial genomic libraries of two Sorex shrews, S. unguiculatus and S. caecutiens, and examined their allelic variation. All loci showed high allelic variation ranging from 15 to 19 alleles and all but one locus conformed to Hardy–Weinberg expectations in the species where the loci were isolated. Cross-species amplifications showed that all primers derived from S. unguiculatus were useful for S. caecutiens, while among primer sets derived from S. caecutiens only one was useful for S. unguiculatus. Accordingly, at least five microsatellite markers were useful in S. caecutiens and three in S. unguiculatus.     

Sequences of the cDNAs and genomic DNAs encoding the S1, S7, S8, and Sf alleles from almond, Prunus dulcis

Partial genomic and cDNA sequences of the RNase alleles S1, S7, S8 and Sf were obtained from Prunus dulcis cvs ’Ne Plus Ultra’, ’Ferragnes’ and ’Nonpareil’ 15–1, and IRTA Selection 12–2. Total DNA was extracted from leaves, and cDNA was prepared from total RNA extracted from styles. The partial cDNA sequences of the S1 allele from ’Ferragnes’, and the S7 and S8 alleles from ’Nonpareil’ 15–1, matched those reported in the literature for the alleles Sb, Sc and Sd respectively. The sequences of the S1, S7, S8 and Sf alleles found in genomic DNA contained introns of 562, 1,530, 2,208 and 689 bp respectively. The exon/intron splice junction sites of all alleles followed the GT/AG consensus sequence rule, and the sequences were found to be highly conserved. Received: 18 October 2000 / Accepted: 8 March 2001     

Plant regeneration from protoplasts of Solanum species with potential agricultural value (S. hjertingii,S. polyadenium,S. capsicibaccatum)

Protoplasts have been isolated from three tuber-bearing Solanum species, S. hjertingii, S. polyadenium and S. capsicibaccatum, that are sexually incompatible with S. tuberosum, but possess potentially useful characters. For isolating protoplasts from leaves of in vitro shoot cultures of S. hjertingii and S. capsicibaccatum growth was improved by including silver thiosulfate in the medium. However, for S. polyadenium, leaves of pot-grown plants were the best source for protoplasts. Following protoplast division and culture, plants were regenerated from protoplasts of each of the species. The pattern of chromosome variation in regenerants was similar to that observed for other diploid and tetraploid Solanum species. The results indicate that it should be possible to introduce the potentially useful germplasm from these wild species into somatic hybrids with S. tuberosum by protoplast fusion.Abbreviations STS silver thiosulfate - BAP benzylaminopurine - GA₃ gibberellic acid - NAA naphthalene acetic acid - IAA indole-3-acetic acid     

Chromatographic and Biological Screening of Chosen Species of Schisandraceae Family: Schisandra chinensis,S. rubriflora,S. sphenanthera,S. henryi and Kadsura japonica

HPLC and TLC profiling was carried out for leaf and fruit extracts of five Schisandraceae species: Schisandra chinensis, S. rubriflora, S. spehenanthera, S. henryi and Kadsura japonica. HPLC measurements confirmed presence of lignans and phenolic compounds in fruits and leaves of all tested species. The most abundant in lignans was S. chinensis fruit extract in which 15 compounds were detected (e. g.: schisandrol A, schisanhenol, γ-schisandrin, gomisin N). The effect-directed detection, i. e., TLC-direct bioautography against Bacillus subtilis, showed exceptionally high activity for S. chinensis and S. rubriflora fruit extracts. On the other hand, TLC-DB enzyme tests (α-glucosidase, lipase, tyrosinase and acetylcholinesterase (AChE) inhibition assays) showed that all fruit and leaf extracts have ability to inhibit the above-mentioned enzymes (except for the K. japonica fruit). The leaf extracts showed much stronger antioxidant activity than the fruit ones, which were assessed and compared using both TLC-direct bioautography and spectrophotometric measurements based on ABTS, DPPH and FRAP tests.     

Sequence Analysis of 18S-28S rRNA Spacer Regions from Saccharomyces kunashirensis, S. martiniae, S. rosinii, and S. transvaalensis

Sequences of two internal transcribed spacer regions between 18S and 28S rRNA for recently described yeasts species, Saccharomyces kunashirensis, S. martiniae, S. rosinii, and S. transvaalensis, were determined to assess their phylogenetic relationship to the other Saccharomyces species. In the two phylogenetic trees constructed by the neighbor-joining method, independent branches reflected that delimitation of the four new species was valid. Received: 17 June 1998 / Accepted: 7 August 1998     

A newSalmonella type (S. Benguella)

Summary A newSalmonella typeS. benguella, with the antigenic formula 40: b:z ₆, is described. The culture was isolated together withS. meleagridis from fish-meal of Portuguese Angola origin.     

Functional characterization of sex pheromone receptors in Spodoptera frugiperda,S. exigua,and S. litura moths

Moths possess an extremely sensitive and diverse sex pheromone processing system, in which pheromone receptors (PRs) are essential to ensure communication between mating partners. Functional properties of some PRs are conserved among species, which is important for reproduction. However, functional differentiation has occurred in some homologous PR genes, which may drive species divergence. Here, using genome analysis, 17 PR genes were identified from Spodoptera frugiperda, S. exigua, and S. litura, which belong to 6 homologous groups (odorant receptor [OR]6, 11, 13, 16, 56, and 62); of which 6 PR genes (OR6, OR11, OR13, OR16, OR56, and OR62) were identified in S. frugiperda and S. exigua, and 5 PR genes were identified in S. litura, excluding OR62. Using heterologous expression in Xenopus oocytes, we characterized the functions of PR orthologs including OR6, OR56, and OR62, which have not been clarified in previous studies. OR6 orthologs were specifically tuned to (Z,E)-9,12-tetradecadienyl acetate (Z9,E12-14:OAc), and OR62 orthologs were robustly tuned to Z7-12:OAc in S. frugiperda and S. exigua. The optimal ligand for OR56 was Z7-12:OAc in S. frugiperda, but responses were minimal in S. exigua and S. litura. In addition, SfruOR6 was male antennae-specific, whereas SfruOR56 and SfruOR62 were male antennae-biased. Our study further clarified the functional properties of PRs in 3 Spodoptera moth species, providing a comprehensive understanding of the mechanisms of intraspecific communication and interspecific isolation in Spodoptera.     

A newSalmonella type (S. Haarlem)

Summary A newSalmonella type (S. haarlem) with the antigenic formula 9,12: z: e,n,x, is described. *** DIRECT SUPPORT *** A1365006 00016     

A set of polymorphic SSR loci for subarctic willow (Salix lanata,S. lapponum and S. herbacea)

We report the development of seven microsatellite markers from a library enriched with [CA]₁₅, [GA]₁₅, [AAG]₈ and [ATG]₈. The library was constructed from Icelandic Salix lanata leaf DNA, and seven highly polymorphic single locus products were tested on a range of Scottish and Icelandic S. lanata samples. These seven loci also amplify polymorphic products when tested for cross amplification in a range of Scottish and Icelandic samples of two other subarctic willows (Salix lapponum and Salix herbacea). The microsatellites developed in this study will provide the framework for comparisons with other ecosystem components, and will ultimately inform conservation strategies.     

Seven newSalmonella types

Seven hitherto unrecognizedSalmonella types are described.S.leiden (13, 22, 36 : z 38),S. enschede (35 : z 10 : l,w) andS. hillegersberg (9, 46 : z 35 : 1,5) were isolated from patients in the Netherlands.S. lawra (44 : k : e,n,z 15) was isolated from a healthy student in Ghana.S. overvecht (30 : a : 1,2),S. woerden (17 : c : z 39) andS. odijk (30 : a : z 39) were isolated from animals kept in zoological gardens in the Netherlands.     

Nonsense suppression in Schizosaccharomyces pombe: the S. pombe Sup3-e tRNASerUGA gene is active in S. cerevisiae

Summary The gene encoding the efficient UGA suppressor sup3-e of Schizosaccharomyces pombe was isolated by in vivo transformation of Saccharomyces cerevisiae UGA mutants with S. pombe sup3-e DNA. DNA from a clone bank of EcoRI fragments from a S. pombe sup3-e strain in the hybrid yeast vector YRp17 was used to transform the S. cerevisiae multiple auxotroph his4-260 leu2-2 trp1-1 to prototrophy. Transformants were isolated at a low frequency; they lost the ability to grow in minimal medium after passaging in non-selective media. This suggested the presence of the suppressor gene on the non-integrative plasmid. Plasmid DNA, isolated from the transformed S. cerevisiae cells and subsequently amplified in E. coli, transformed S. cerevisiae his4-260 leu2-2 trp1-1 to prototrophy. In this way a 2.4 kb S. pombe DNA fragment carrying the sup3-e gene was isolated. Sequence analysis revealed the presence of two tRNA coding regions separated by a spacer of only seven nucleotides. The sup3-e tRNA _Ser ^UGA tRNA gene is followed by a sequence coding for the initiator tRNA^Met. The transformation results demonstrate that the cloned S. pombe UGA suppressor is active in S. cerevisiae UGA mutant strains.     

Eight newSalmonella types

Summary Eight newSalmonella types are described.S. houten, 43; Z₄, Z₂₃ :−, was isolated from a frog,S.tuindorp, 43: Z₄, Z₃₂: −,S. vleuten, 44: fg: −,S.zeist, 18: Z₁₀: Z₆, andS.soesterberg. 21: Z₄, Z₂₃: −, were isolated from lizards. S. kralingen (8), 20: y: Z₆, was isolated from peanuts andS. assen, 21: a: −, was isolated from a swine. All these strains were isolated in the Netherlands. S.mim, 13, 22: a: 1,6, was isolated from a rat in Ghana.     

Isolation,identification and characterization of Staphylococcus sp. from Indian ethnic fermented fish product

Staphylococci from Sheedal of Northeast India was isolated, identified and characterized. All the isolated staphylococci were found to be coagulase negative. Based on the rpoB gene sequences followed by analysis using NCBI-BLAST software, seven species of Staphylococcus namely, S. piscifermentans, S. condimenti, S. arlettae, S. sciuri, S. warneri, S. nepalensis and S. hominis were recognized. Phylogenetic analyses revealed three major cluster groups. All the seven Staphylococcus showed their NaCl tolerance from 2 to 8%. No species was able to grow at 55°C. Except S. arlettae and S. sciuri, all the isolated staphylococcal species exhibited growth at pH 4–8. No isolated species was able to ferment mannitol, sucrose and arabinose. All the species exhibited moderate to maximum proteolytic and lipolytic activities. All the seven species were found to be sensitive to the antibiotics, namely, erythromycin, norfloxacin, ampicillin, streptomycin and vancomycin, whereas all were resistant to co-trimoxazole. Only S. piscifermentans was found antagonist to Salmonella enterica, Escherichia coli and Bacillus subtilis, although the clear zone was minimal. All the staphylococcal species except S. arlettae and S. sciuri exhibited hydrophobicity ranging from 25 to 66%. The observed characteristics of isolated Staphylococci from Sheedal revealed their role in fish fermentation.     

Sequence Analysis of Small Cryptic Plasmids Isolated from Selenomonas ruminantium S20

Two small cryptic plasmids designated pONE429 and pONE430 were isolated from a rumen bacterium, Selenomonas ruminantium S20. The complete sequence of pONE429 was 2100 bp and contained one open reading frame (ORF) of 201 amino acids. The sequence of pONE430 had 1527 bp and one ORF of 171 amino acids with the similarity of replication protein (Rep protein) of pOM1, pSN2, and pIM13 isolated from Butyrivibrio fibrisolvens, Staphylococcus aureus, and Bacillus subtilis, respectively. In these plasmids, the upstream nucleotide sequence of Rep protein had the conserved nucleotides which could be double-strand origin (DSO) of rolling circle replication (RCR) mechanism. The plasmids of pONE429, pONE430, pJJMI, pJDB21, and pS23 were isolated from S. ruminantium strains and had similar regions that were located within a <450-bp nucleotide. These similar regions may be the location that was recognized by the host strain, S. ruminantium. Received: 10 July 1998 / Accepted: 11 September 1998     

Enterotoxigenicity and invasiveness of Salmonella species

A large number of enterotoxigenic strains was encountered in a group 56 Salmonella cultures belonging to 8 species viz., S. alachua, S. anatum, S. dublin, S. enteritidis, S. hindmarsh, S. newport, S. typhimurium, S. weltevreden, and 5 serotypes of S. arizona (16:z₄:-; 48:1,v:z₅₆; 53:z₅₂:z₅₃; 60:r:z; 61:i:z₅₃). These cultures were isolated mainly from humans and animals suffering from gasteroenteritis. The enterotoxigenic (diarrhoeagenic) Salmonella cultures possess capacities for both skin permeation and epithelial penetration (invasiveness). Preliminary characterization revealed that Salmonella enterotoxin is a heatlabile protein of high molecular weight. It is suggested that enterotoxigenic and invasive propeties play a vital role in the pathogenesis of Salmonella diarrhoea.