Taxonomic study of twoProteocephalus secies (Cestoda: Proteocephalidae) parasitising coregonid fishes: the synonymy ofP. fallax La Rue, 1911 withP. exiguus La Rue, 1911

Type-specimens (syntypes) and comparative material of twoProteocephalus species,P. exiguus La Rue, 1911 andP. fallax La Rue, 1911, parasitic in coregonid fishes from the Holarctic Region, were evaluated taxonomically. Morphological and biometrical study revealed conspecificity of the taxa.P. fallax is therefore, considered to be a synonym ofP. exiguus. Supplementary information is added to the species diagnosis ofP. exiguus and a lectotype is designated herein.     

A Short Region from the LEU2 Gene of Saccharomyces cerevisiae Functions as an ARS in the Yeast Saccharomyces exiguus Yp74L-3

We examined the autonomously replicating sequence (ARS) activity of some fragments derived from the LEU2 region of Saccharomyces cerevisiae onto Saccharomyces exiguus Yp74L-3. A DNA fragment functioning as an ARS in S. exiguus, but not in S. cerevisiae, was shown to exist. The ARS activity for S. exiguus was reduced by the 2-μm plasmid origin of S. cerevisiae when both elements coexisted on a single circular plasmid. Analysis of ARS activity with the PCR products from the fragment revealed that the ARS-acting sequence was located in the 3′-terminal area of the transcribed region of the LEU2 gene of S. cerevisiae. It is suggested that the ARS recognition system in S. exiguus is significantly different from that of S. cerevisiae. Received: 16 June 1998 / Accepted: 3 August 1998     

The Autonomously Replicating Sequence (ARS) of the Yeast Saccharomyces exiguus Yp74L-3

Fragments containing ARSes were cloned from the genomic DNA of the yeast Saccharomyces exiguus Yp74L-3, and the essential regions for ARSes were restricted for these fragments. Mapping studies of ARS-acting sequences in one of these fragments suggested that S. exiguus recognizes a sequence as an ARS that is different from that recognized by Saccharomyces cerevisiae. Two ARS essential regions of S. exiguus were sequenced, and an ARS core consensus sequence of S. exiguus was deduced to be MATTAMWAWWTK. This sequence differs significantly from that of S. cerevisiae in two positions, suggesting that these nucleotide substitutions cause the difference in the ARS-recognition modes between S. exiguus and S. cerevisiae. Received: 3 August 1998 / Accepted: 18 September 1998     

Taxonomical classification of yeasts isolated from kefir based on the sequence of their ribosomal RNA genes

Yeast strains present in 10 samples of kefir of different commercial and domestic origins have been isolated and classified taxonomically on the basis of the internal transcribed sequences (ITS) of their ribosomal RNA genes. A total of 18 yeast strains representing 10 different species have been characterized. Of the three commercial kefir samples analyed, two contained the well characterized yeast Kluyveromyces lactis while no yeast was found in the other one. A broader spectrum of yeast species was found among the home-made kefir samples, of which Issatchenkia orientalis, Saccharomyces unisporus, Saccharomyces exiguus and Saccharomyces humaticus were the most representative species.     

Saccharomyces barnetti andSaccharomyces spencerorum: two new species ofSaccharomyces sensu lato (van der Walt)

In the course of a study of DNA base sequence homology, 19 strains labelled asSaccharomyces exiguus, its imperfect state,Candida holmii, orC. milleri were examined. Results confirmed the separation ofC. milleri as a separate species. The remaining strains can be divided into three distinct groups of genomic relatedness. The type cultures ofS. exiguus andC. holmii form a cluster with 10 other strains showing variable reassociation values ranging from 100 to 40%. The remaining four strains comprise two separate species showing no nucleotide relatedness between themselves nor to either theS. exiguus complex or toC. milleri. Physiological analyses demonstrate that it is possible to separate these four taxa on the basis of a few simple tests. The two new species are described respectively asSaccharomyces barnetti honoring James Barnett in recognition of his invaluable work in the field of yeast taxonomy andSaccharomyces spencerorum in honor of J.F.T. and Dorothy M. Spencer who have made innumerable contributions to the genetics and biotechnological application of yeasts.     

Saccharomyces bulderi sp. nov., a yeast that ferments gluconolactone

An unknown yeast species was isolated from maize silage and was determined to be novel on the basis of morphological and physiological characteristics, nucleotide sequence of domain D1/D2 of LSU rDNA and from its electrophoretic karyotype. The name for the proposed new species is Saccharomyces bulderi Middelhoven, Kurtzman et Vaughan-Martini (type strain CBS 8638, NRRL Y-27203, DBVPG 7127). S. bulderi is closely related to S. barnettii and S. exiguus from which it can be distinguished by having a double vitamin requirement of biotin and thiamine and by no or slow aerobic growth on raffinose, a sugar that on the contrary is fermented rapidly. Gluconolactone is rapidly fermented with ethanol, glycerol and carbon dioxide being the main products.     

Molecular Cloning and DNA Analysis of the Orotidine-5′-Phosphate Decarboxylase Gene from the Yeast Saccharomyces exiguus Yp74L-3

The orotidine-5′-phosphate decarboxylase gene of Saccharomyces exiguus Yp74L-3 was cloned as a DNA fragment complementing a ura4 mutation of this yeast. The coding region of the gene is 807 bp in length, and represents 68.7% similarity to the corresponding gene of S. cerevisiae (URA3). The cloned URA4 gene was shown to be located on the 790-kbp Chromosome (chr) VIII of S. exiguus Yp74L-3. The neighbor-joining phylogenetic tree based on the orotidine-5′-phosphate decarboxylase coding sequences indicates that S. exiguus Yp74L-3 is closely related to Kluyveromyces yeasts, as well as to a S. cerevisiae laboratory strain. Received: 4 February 2000 / Accepted: 3 July 2000     

The sourdough microflora. Interactions between lactic acid bacteria and yeasts: metabolism of amino acids

Co-culture of Lactobacillus brevis subsp. lindneri or L. plantarum with Saccharomyces cerevisiae or S. exiguus from sourdough did not modify the yield of the yeasts but gave higher growth rates and final yields of both lactic acid bacteria (LAB) than in their respective mono-cultures. Co-cultures of L. brevis subsp. lindneri with S. cerevisiae or S. exiguus in a medium without valine or leucine, which are essential for growth of the LAB, led to growth of the LAB due to excretion of these amino acids by the yeasts.The authors are with the Institute of Dairy Microbiology, Faculty of Agriculture, University of Perugia, Via S. Costanzo, 06100 Perugia, Italy     

Electrophoretic karyotyping as a taxonomic tool in the genusSaccharomyces

Summary The electrophoretic karyotypes of strains of the ten species of the yeast genusSaccharomyces (sensu Vaughan-Martini & Martini 1992) were determined by the CHEF (contour-clamped homogeneous electric field) system of pulsed field gel electrophoresis. The number of bands was found to vary from 6 to 17 and the calculated molecular weights of haploid genomes ranged from 7.9 to 14.6 Mbp. The type strains ofS. exiguus and the four species of theSaccharomyces sensu stricto complex (S. bayanus, S. cerevisiae, S. paradoxus andS. pastorianus) have genomes comprised of chromosomes of all three size classes: light (< 500 kb), medium (500–1000 kb) and heavy (> 1,000 kb).Saccharomyces kluyveri DNA has only heavy bands, while the remaining species exhibit medium and heavy chromosomes. When more than one strain of each species was examined, it was seen that while the speciesS. bayanus, S. castellii, S. cerevisiae, S. kluyveri, S. paradoxus andS. pastorianus showed uniform karyotypes,S. dairensis, S. exiguus, S. servazzii andS. unisporus comprise heterogeneous taxa.     

Physical Characterization of the Chromosomal DNA of the Yeast Saccharomyces exiguus

The number of chromosomes in the yeast Saccharomyces exiguuswas determined to be thirteen by two-dimensional pulsed-fieldgel electrophoresis. The thirteen chromosomes ranged in DNAsize from 520 to 2,600 kbp, with a total length of approximately14 Mbp. Numbers I to XIII were assigned to the chromosomes indecreasing order of DNA length. Southern hybridization analysisusing total DNAs from S. exiguus and S. cerevisiae as probesshowed that there was no significant homology between the chromosomalDNAs of the two species, except in the case of the chromosomalDNA that included rDNA. When rDNA and genes LEU2, TRP1, URA3and HO of S. cerevisiae were used as hybridization probes, itwas apparent that S. exiguus had DNA sequences homologous tothe rDNA and to the LEU2 and HO genes. In S. exiguus, rDNA-likeand LEU2-like DNAs were located on chromosomes I and IX, respectively,and HO-like DNA was located on chromosome VI or VII. (Received May 17, 1993; Accepted July 15, 1993)     

Diapause of copepods as an element for stabilizing the parasite system of some fish helminths

The analysis of infection dynamics of copepods, which are the intermediate hosts of three helminth species of freshwater fishes (Triaenophorus crassus, Proteocephalus exiguus and P. percae), has shown that diapause in the life cycle of the copepods is favourable for preserving the infection in the waterbody until physiological prerequisites for successful infection of the final host are acquired. The peculiarity of a copepod's life cycle may determine the strategy of a parasite in its preimaginal phase as a waiting stage, and the duration of the residence of helminth larvae in copepods that have an obligatory diapause, is one of the elements the provide stability in parasite systems.     

The sourdough microflora. Interactions between lactic acid bacteria and yeasts: metabolism of carbohydrates

Interactions betweenLactobacillus brevis subsp.lindneri CB1,L. plantarum DC400,Saccharomyces cerevisiae 141 andS.exiguus M14 from sourdoughs were studied in a co-culture model system using a synthetic medium. The lack of competition for maltose whenS.exiguus M14 was present in co-culture with each of the lactic acid bacteria (LAB) enhanced the bacterial cell yield and lactic and acetic acid production.L.brevis subsp.lindneri CB1 resting cells hydrolysed maltose and accumulated glucose in the medium, allowing the growth of maltose negative yeast.S.cerevisiae 141 competed greatly with each of the LAB for glucose and only withL.plantarum DC400 for fructose, causing a decrease in the bacterial cell number and in acid production. As a result of the glucose and fructose availability after the invertase activity of both yeasts,L.plantarum DC400 grew optimally in the presence of sucrose as a carbon source. All of the interactions indicated were confirmed by studying the behaviour of the co-cultures in wheat flour hydrolysate.     

Identification of the Orotidine-5′-Phosphate Decarboxylase Gene and Development of a Transformation System in the Yeast Saccharomyces exiguus Yp74L-3

To investigate the uracil biosynthetic pathway of the yeast Saccharomyces exiguus Yp74L-3, uracil auxotrophic mutants were isolated. Using conventional genetic techniques, four mutant genes concerned in uracil biosynthesis were identified and denoted as ura1, ura2, ura3, and ura4. Mutations in the URA3 and URA4 genes were specifically selected with 5-fluoroorotic acid (5-FOA). Vector plasmids containing the URA3 gene and an autonomously replicating sequence (ARS) of S. cerevisiae produced sufficient amounts of Ura⁺ transformants from the ura4 mutant of S. exiguus. This fact indicates that the S. exiguus URA4 gene encodes orotidine-5′-phosphate decarboxylase (OMP decarboxylase) and demonstrates that vector plasmids for S. cerevisiae are also usable in S. exiguus.     

A re-appraisal of two members of the genus Notholca from the Andes,with a note on the fine structure of the lorica of N. foliacea (Ehrenberg)

Rotifers described from the Andes by Murray (1913) and De Beauchamp (1939) as Notholca foliacea (Ehrenberg) are reviewed and re-assessed as Notholca walterkostei De Paggi, 1982. They are compared with N. foliacea and details of the lorica of this species seen with the scanning electron microscope are presented.     

Review of the Indo-Pacific gobiid fish genus Lubricogobius, with description of a new species and a new genus for L. pumilus

The gobiid genus Lubricogobius Tanaka, distinctive in being scaleless, without sensory pores, but with sensory papillae on the head, consists of three small species (largest, 33.7 mm SL): the yellow L. exiguus Tanaka, type species, from Japan, with probable records from Taiwan and New Caledonia; L. ornatus Fourmanoir, orange with blue lines on the head, described from Vietnam, recorded from the Arafura Sea, and the range here extended to the Timor Sea and Northwest Shelf of Western Australia, Ryukyu Islands, and to New Caledonia; and L. dinah, a new species from Papua New Guinea and the Ryukyu Islands, white dorsally and abruptly orange-yellow ventrally, with 10–11 dorsal soft rays. L. pumilus Larson and Hoese, known from one 14.5-mm specimen from off Somalia, is referred to the new genus Larsonella. The new genus differs from Lubricogobius in having scales posteriorly on the body, a more slender body, depressed head, no pelvic frenum, the snout longer than orbit diameter, and a different pattern of sensory papillae on the head. Received: September 30, 1999 / Accepted: June 9, 2000     

Interaction between lactic acid bacteria and yeasts in sour-dough using a rheofermentometer

Rheofermentometer assays were used to characterize the leavening of sour-doughs produced using species of lactic acid bacteria (LAB) and yeasts, alone or in combination. Saccharomyces cerevisiae 141 produced the most CO₂ and ethanol whereas S. exiguus M14 and Lactobacillus brevis subsp. lindneri CB1 contributed poorly to leavening and gave sour-doughs without porosity. In comparison with that seen in sour-dough produced with yeast alone, yeast fermentation with heterofermentative LAB present was faster whereas that with homofermentative LAB (L. plantarum DC400, L. farciminis CF3) present was slower and produced more CO₂. Combining L. brevis subsp. lindneri CB1 with S. cerevisiae 141 decreased bacterial cell numbers and souring activity. However, addition of fructose to the sour-dough overcame these problems as well as activating S. cerevisiae 141.The authors are with the Institute of Dairy Microbiology, Faculty of Agriculture, University of Perugia, S. Costanzo, 06126 Perugia, Italy     

A New Pollen Type in the Winteraceae

Pollen of Zygogynum balansae and Z. pomiferum is described with the aid of the scanning electron microscope. Other members of the primitive ranalean family Winteraceae hitherto investigated have pollen in permanent tetrads, but these two species have solitary grains. Sculpturing is similar on the distal and proximal faces of each grain, excluding the distal apertural zone. The exine pattern resembles that on distal faces of tetrads of other Zygogynum species investigated, including Z. bailloni, the pollen of which is illustrated. Although Zygogynum and Exospermum have rather similar pollen, with sculpturing differing from other Winteraceae in consisting of a less coarse reticulum, pollen of Exospermum stipitatum is clearly distinct from the Zygogynum species investigated, as seen in the scanning electron microscope. It is not clear whether or not the solitary pollen type has evolved from the tetrad form.