Candida easanensis sp. nov., Candida pattaniensis sp. nov. and Candida nakhonratchasimensis sp. nov., three new species of yeasts isolated from insect frass in Thailand

Six strains of anamorphic yeasts isolated from insect frass collected in several regions of Thailand were assigned to the genus Candida based on the conventional taxonomic criteria used for yeast classification. These strains have Q-7 as the major ubiquinone and are suggested to have close relationships to the genus Pichia. Three strains, ST-225, ST-228 and ST-229, have identical nucleotide sequences in the D1/D2 domain of 26S rDNA and are closely related to Pichia japonica, but differ by six nucleotides (1.1% ) from this species. These three strains are considered to represent a single new species, which is described as Candida easanensis sp. nov. Two strains, ST-311 and ST-320, have identical sequences in the D1/D2 domain and resemble Pichia veronae and Pichia fabianii but differ from them by nine nucleotides (1.6%) in D1/D2 sequences. The two strains are described as Candida pattaniensis sp. nov. The remaining strain, ST-37, is related to Pichia americana and Pichia bimundalis but differs by six(1.1%) and seven (1.2%) nucleotides from these species, respectively. This strain is described as Candida nakhonratchasimensis sp. nov.     

Heat Resistance Studies on Yeast spp. Causing Spoilage in Soft Drinks

A test method to investigate the heat resistance of yeasts has been developed. The method was used to study the heat resistance of 120 yeast strains, representative of the fungal flora in soft drinks and certain acid food products: 35 asporogenous yeast strains ( Brettanomyces, Candida, Kloeckera, Rhodotorula and Torulopsis ) and 85 ascomycetous strains ( Debaryomyces, Hansenula, Kluyveromyces, Lodderomyces, Pichia, Saccharomyces and Saccharomycopsis ) were tested. Generally, asporogenous yeasts were found to be less heat resistant than ascomycetous types. The genus Saccharomyces showed the highest heat resistance, especially strains of the species Sacch. cerevisiae and Sacch. chevalieri . For an evaluation of the practical implications of these results additional studies on environmental factors influencing the heat resistance of ascomycetous yeast species are required.     

Quantitative and qualitative seasonal changes in the microbial community from the phyllosphere of sugar beet (Beta vulgaris)

Bacteria, yeasts and filamentous fungi colonizing immature, mature and senescing primary leaves of field grown Beta vulgaris (sugar beet) were analysed over a complete growing season. Greatest microbial numbers were detected on senescing primary leaves and these numbers increased over most of the season. The number of colonizers detected on mature leaves was found to be stable over most of the study.Filamentous fungi and yeasts were identified to the genus level and the communities found to have greatest diversity during the summer months. There was no consistent pattern of diversity according to leaf type. Two genera of filamentous fungi, Cladosporium and Alternaria and two yeast genera, Cryptococcus and Sporobolomyces were the most numerous fungal populations isolated. Only 8 filamentous fungi and 3 yeast genera were commonly isolated on PDA (potato dextrose agar).Bacterial strains (1236) were isolated on Tryptic Soy Broth (TSB) agar and identified to species, or in some cases sub-species level, by analysis of their fatty acid methyl ester (FAME) profiles. Isolated bacteria were grouped into 78 named and 37 unnamed species clusters. Greatest number of bacterial species were isolated from young plants and leaves, sampled during the autumn months. Bacterial community diversity was lowest in mid-summer and winter months. Pseudomonas was the most commonly isolated genus and Erwinia herbicola the most common species. P. aureofaciens was the only species isolated from soil that was also isolated from the phyllosphere of B. vulgaris throughout the season.     

Quantification of different yeasts associated with the bark beetle,Ips typographus,during its attack on a spruce tree

There were different amounts and types of yeasts associated with individuals ofIps typographus spruce bark beetles during different phases of their attack on a healthy spruce tree. The yeasts were isolated on Sabouraud agar medium in order to identify them and estimate their numbers.Hansenula holstii andCandida diddensii type yeasts were most frequently isolated. The increase in number of these two yeast types probably accounted for most of the total yeast increase found during the later attack phases of the bark beetles. Lesser amounts ofHansenula capsulata, Pichia pinus, Candida nitratophila, and twoCryptococcus type yeasts were also found.     

Gut-associated yeast in bark beetles of the genus Dendroctonus Erichson (Coleoptera: Curculionidae: Scolytinae)

Scolytine bark beetles are the most destructive pests of conifers; they sometimes aggregate in such large numbers that they actually kill their hosts. They maintain close relationships with yeasts and fungi, in particular those that are assumed to aid in digestive, detoxification processes and pheromone production. In this study, 403 yeast strains were isolated from the guts, ovaries, eggs and frass of nine bark beetle species in the genus Dendroctonus Erichson. The beetles were collected from 10 conifer species at 34 locations in Mexico, Guatemala and the USA. Yeast identification was based on partial DNA sequences from 18S rDNA, 26S rDNA and internal transcribed spacer (ITS1), as well as morphological and physiological characteristics. A combined phylogenetic analysis delimited 11 clades with sequences similar to Candida arabinofermentans , C. ernobii , C. membranifaciens (including C. lessepsii , Pichia mexicana and P. scolyti ), C. oregonensis , C. piceae , Kuraishia capsulata (including K. capsulata and K. cf. molischiana ), Pichia americana , P. canadensis , P. glucozyma , P. guilliermondii and an undescribed species of Candida . Nucleotide divergences between the major clades were at least 5% while, with the exception of 30 isolates, yeasts within clades differed from named reference species at fewer than 1% of the nucleotide sites. There do not appear to be obligate relationships between particular yeasts and specific anatomical partitions, nor between particular yeasts and bark beetle species. Some yeasts do appear to be preferentially associated with bark beetles feeding on different conifer genera and therefore host plant defences may limit yeast community diversity in Dendroctonus .  © 2009 The Linnean Society of London, Biological Journal of the Linnean Society , 2009, 98 , 325–342.     

A taxonomic study of soil yeasts

Summary Eighty-four samples of Minnesota soils were collected in the spring of the year. All samples yielded yeasts, and a total of 180 cultures were isolated of which 117 were studied taxonomically. Twenty-five cultures were black yeasts. Approximately one-third of the isolates were spore forming yeasts and members of the oxidative and film forming generaHansenula, Pichia, andDebaryomyces, of which a new species (D. mrakii) has been described. Not a single culture ofSaccharomyces was isolated. Perhaps later in the season when fruit was abundant members of this genus would have been present.The greatest number of imperfects belonged to the genusCandida (32 cultures), althoughPullularia (25 cultures),Rhodotorula (20 cultures), andTorulopsis (16 cultures) were well represented.The cultures ofPullularia darkened very slowly if at all, in the asbence of excess sugar. Under these circumstances they were very similar toT. pullulans.A single culture ofTrichisporon cerebriformis was isolated. The culture isolated formed both blastospores and arthrospores.I would like to express my appreciation to Dr.C. E. Skinner of Washington State College, for his help and guidance throughout this entire problem. I would also like to thank Dr.E. M. Mark, of the University of California, Berkeley, for the privilege of working under his direction, and in his laboratory during the study.     

Les levures des Drosophiles de savane d'Afrique intertropicale (Savanes De Lamto,Côte d'ivoire)

Yeasts from crops of Drosophilidae (154 flies belonging to 22 species) of Lamto savannas (Ivory Coast) were studied. 50 yeast strains (belonging to 19 species) were isolated and studied. The most frequently isolated yeasts wereHanseniaspora andPichia membranaefaciens s.l.; these yeasts ferment and assimilate few of the carbon compounds. Some crops contained until 100 000 yeast cells. In 25% of the flies, these yeasts belonged to several different species.     

Diversity of Soil Yeasts Isolated from South Victoria Land,Antarctica

Unicellular fungi, commonly referred to as yeasts, were found to be components of the culturable soil fungal population in Taylor Valley, Mt. Discovery, Wright Valley, and two mountain peaks of South Victoria Land, Antarctica. Samples were taken from sites spanning a diversity of soil habitats that were not directly associated with vertebrate activity. A large proportion of yeasts isolated in this study were basidiomycetous species (89%), of which 43% may represent undescribed species, demonstrating that culturable yeasts remain incompletely described in these polar desert soils. Cryptococcus species represented the most often isolated genus (33%) followed by Leucosporidium (22%). Principle component analysis and multiple linear regression using stepwise selection was used to model the relation between abiotic variables (principle component 1 and principle component 2 scores) and yeast biodiversity (the number of species present at a given site). These analyses identified soil pH and electrical conductivity as significant predictors of yeast biodiversity. Species-specific PCR primers were designed to rapidly discriminate among the Dioszegia and Leucosporidium species collected in this study.     

Microflora of partially processed lettuce

Bacteria, yeasts, and molds isolated from partially processed iceberg lettuce were taxonomically classified. The majority of bacterial isolates were gram-negative rods. Pseudomonas, Erwinia, and Serratia species were commonly found. Yeasts most frequently isolated from lettuce included members of the genera Candida, Cryptococcus, Pichia, Torulaspora, and Trichosporon. Comparatively few molds were isolated; members of the genera Rhizopus, Cladosporium, Phoma, Aspergillus, and Penicillium were identified.     

Yeasts in some Netherlands soils

Summary In 12 Netherlands soil samples, in two successive analyses carried out in the autumn of 1952, the author has isolated up to 105 cultures of yeasts to be classified under the following species:Kloeckera apiculata, Torulopsis inconspicua, Torulopsis pulcherrima, Candida Guilliermondii, Rhodotorula mucilaginosa, Aureobasidium pullulans, Hansenula saturnus, Hansenula californica, Saccharomyces mangini, Saccharomyces ellipsoideus andPichia membranefaciens. In all the soils examined the quantity of yeasts found has been very small. The results obtained do not offer evidence for any relationship between manurial treatment and species and number of yeasts occurring. Sporeforming yeasts were less numerous than non-sporeforming ones.

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Riassunto Da 12 terreni Olandesi, in due successive analisi, eseguite nell'autunno 1952, l'A. riferisce di avere isolato ben 105 colture pure di lieviti da riportare alle seguenti specie:Kloeckera apiculata, Torulopsis inconspicua, Torulopsis pulcherrima, Candida Guilliermondii, Rhodotorula mucilaginosa, Aureobasidium pullulans, Hansenula saturnus, Hansenula californica, Saccharomyces mangini, Saccharomyces ellipsoideus ePichia membranefaciens. In tutti i terreni esaminati la quantità di lieviti riscontrati è stata molto esigua; non è stata posta in evidenza alcuna influenza in rapporto alle diverse concimazioni ed è stata rilvata la scarsissima diffusione delle specie sporigene su quelle incapaci di formare spore.

     

The yeasts of British fresh sausage and minced beef

Three hundred and eighty three yeasts isolated from samples of unsulphited or sulphited sausages and skinless sausages and minced beef were characterized in detail.Debaryomyces hansenii was the most commonly isolated yeast from most samples followed byCandida zeylanoides andPichia membranaefaciens. The presence of sulphite in sausages did not appear to affect the numbers and range of yeasts present but did affect their relative proportions. A survey of one factory showed that meat intended for sausage production and equipment harboured the same range of yeasts that are found in the finished products.     

Linear mitochondrial DNAs of yeasts: frequency of occurrence and general features.

In most yeast species, the mitochondrial DNA (mtDNA) has been reported to be a circular molecule. However, two cases of linear mtDNA with specific termini have previously been described. We examined the frequency of occurrence of linear forms of mtDNA among yeasts by pulsed-field gel electrophoresis. Among the 58 species from the genera Pichia and Williopsis that we examined, linear mtDNA was found with unexpectedly high frequency. Thirteen species contained a linear mtDNA, as confirmed by restriction mapping, and labeling, and electron microscopy. The mtDNAs from Pichia pijperi, Williopsis mrakii, and P. jadinii were studied in detail. In each case, the left and right terminal fragments shared homologous sequences. Between the terminal repeats, the order of mitochondrial genes was the same in all of the linear mtDNAs examined, despite a large variation of the genome size. This constancy of gene order is in contrast with the great variation of gene arrangement in circular mitochondrial genomes of yeasts. The coding sequences determined on several genes were highly homologous to those of the circular mtDNAs, suggesting that these two forms of mtDNA are not of distant origins.     

Yeast communities of the cactusPilosocereus arrabidae as resources for larval and adult stages ofDrosophila serido

The feeding behavior ofDrosophila serido on the yeast communities of necrotic stem tissue ofPilosocereus arrabidae were studied in a sand dune ecosystem of Rio de Janeiro, Brazil. The prevalence of cactophilic yeasts includingPichia barkeri, Candida sonorensis andGeotrichum sp. in the crops and external surfaces ofD. serido reflected its association with the cactus habitat. The effective number of yeasts vectored on the surface of flies was higher than that in the crops. Also overlap between the yeasts from stems and from crops was partial suggesting selective feeding by the flies in the substrates visited. The females had a higher effective number of yeast species and a lower similarity than males with the yeast community ofP. arrabidae. This was probably related to the search for oviposition sites by females. The presence ofPichia thermotolerans-like andPichia amethionina varpachycereana in the flies, but not inP. arrabidae stems, indicated thatD. serido was not limited to this cactus species. The larvae and adults lived in different patches with the adults feeding in patches with higher yeast species richness. The larvae had a narrower feeding niche and higher overlap withP. arrabidae, and preferredP. barkeri andPichia cactophila as food. Adult flies fed on patches with the most frequent yeasts except forP. cactophila. Pichia caribaea was found in higher frequency in the adult crops than in the stems. Our data suggested that there was food selection and diet partitioning between adult and larval stages ofD. serido.     

Three new thermotolerant methylotrophic yeasts, Candida krabiensis sp. nov., Candida sithepensis sp. nov., and Pichia siamensis sp. nov., isolated in Thailand

Five strains of thermotolerant methylotrophic yeasts isolated in Thailand were found to represent three new species in the genera Pichia and Candida, based on phylogenetic analysis of D1/D2 domain of 26S rDNA, in addition to the morphological, physiological, biochemical and chemotaxonomic characterization. Three strains, FS96 and FS101 from flowers and M02 from tree flux, were characterized by ubiquinone Q7, multilateral budding, and the formation of hat-shaped ascospores that are liberated at maturation. These strains showed identical nucleotide sequences in the D1/D2 domain and formed a cluster with Candida thermophila, "Pichia salicis" and Pichia angusta. They differed by 1.9% of nucleotide substitutions from Candida thermophila, the nearest species. They were considered to represent a single new species and are described as Pichia siamensis sp. nov. Two strains, N051 and S023, isolated from soil did not produce ascospores, proliferated by multilateral budding, did not demonstrate urease or DBB color reaction, and lacked sexual stages. These characteristics correspond to the genus Candida. Strains N051 and S023 differed by 2.8% and 1.9% of nucleotide substitutions in the D1/D2 domain from the nearest species, Candida nemodendra and Candida ovalis, respectively, and are considered to represent respective new species. N051 and S023 are described as Candida krabiensis sp. nov. and Candida sithepensis sp. nov., respectively.     

Yeast communities as descriptors of habitat use by the Drosophila fasciola subgroup (repleta group) in Atlantic rain forests

Yeast communities associated with four species of the Drosophila fasciola subgroup (repleta group) in tropical rain forests were surveyed in an abandoned orchard, and rain forest sites of Rio de Janeiro and Ilha Grande, State of Rio de Janeiro, Brazil. Adult flies of Drosophila carolinae, Drosophila coroica, Drosophila fascioloides and Drosophila onca frequently carried Candida colliculosa, Geotrichum sp, Kloeckera apiculata and a Pichia membranaefaciens-like species. The most frequent yeasts in the crop of flies included Candida collicullosa, C. krusei, Pichia kluyveri and a P. membranaefaciens-like species. The physiological abilities and species composition of these yeast communities differed from those of other forest-inhabiting Drosophila. The narrow feeding niches of the fasciola subgroup suggested the use of only part of the substrates available to the flies as food in the forest environment, as noted previously for cactophilic Drosophila serido (mulleri subgroup of the repleta group) in a sand dune ecosystem. The cactophilic yeasts that were isolated have not been previously found in forests. The fasciola subgroup probably used epiphytic cactus substrates as breeding and feeding sites in the forest. The physiological profile of yeasts associated with the fasciola flies was broader than that of yeasts associated with the cactophilic Drosophila serido, suggesting that the fasciola subgroup represents an older lineage from which the South American repleta species evolved.     

Pichia spartinae sp. n. from Louisiana marshland habitats

A new species ofPichia has been described.Pichia spartinae was one of the predominant yeasts isolated from the rhizosphere and tissue of oyster grass,Spartina alterniflora, in the Louisiana marshland. The species occurs in the environment in both the homo- and heterothallic state. This is the first species ofPichia in high densities in an estuarine locality.We wish to express our gratitude to Mr. J. C. Raadschelders for correcting the latin diagnosis.     

Morphological and ecological similarities: wood-boring beetles associated with novel xylose-fermenting yeasts, Spathaspora passalidarum gen. sp. nov. and Candida jeffriesii sp. nov.

Ascomycete yeasts that both ferment and assimilate xylose were reported previously as associates of insects living in woody substrates. Most notable have been reports of Pichia stipitis-like yeasts that are widely associated with the wood-boring beetle, Odontotaenius disjunctus (Coleoptera: Passalidae), in the eastern United States. Our continuing investigation of insect gut yeasts has lead to the discovery of two new xylose-fermenting yeasts that phylogenetic analysis places as sister taxa. The beetle hosts, O. disjunctus and Phrenapates bennetti (Coleoptera: Tenebrionidae), are similar in habitat and appearance, and the presence of similar gut yeasts is an additional common feature between them. Here we describe the new yeast genus Spathaspora, the type species S. passalidarum, and its sister taxon Candida jeffriesii and discuss their natural history, including a comparison with Pichia stipitis, another member of a guild of xylose-fermenting yeasts with similar metabolic traits. In addition a morphologically distinct yeast ascospore type is described for Spathaspora.