Hanseniaspora occidentalis sp. nov.

An ascigerous state was observed in the type strain of Kloeckera occidentalis, which is described as Hanseniaspora occidentalis sp. n.     

Description of Tropicibacter mediterraneus sp. nov. and Tropicibacter litoreus sp. nov.

Four strains (M15∅_3, M17^T, M49 and R37^T) were isolated from Mediterranean seawater at Malvarrosa beach, Valencia, Spain. Together with an older preserved isolate (strain 2OM6) from cultured oysters at Vinaroz, Castellón, Spain, the strains were thoroughly characterized in a polyphasic study and were placed phylogenetically within the Roseobacter clade in the family Rhodobacteraceae. Highest 16S rRNA sequence similarities of the five strains to the types of any established species corresponded to Tropicibacter multivorans (95.8–96.4%), Phaeobacter inhibens (95.9–96.3%) and Phaeobacter gallaeciensis (95.9–96.2%). On the other hand, whole genome (ANI) and protein fingerprinting (MALDI-TOF) data proved: (i) non clonality among the strains, and (ii) the existence of two genospecies, one consisting of strains M15∅_3, M17^T, M49 and 2OM6 and another one consisting of strain R37^T. Phenotypic traits determined allow differentiating both genospecies from each other and from closely related taxa. In view of all data collected we propose to accommodate these isolates in two species as members of the genus Tropicibacter, Tropicibacter mediterraneus sp. nov. (type strain M17^T = CECT 7615^T = KCTC 23058^T) and Tropicibacter litoreus sp. nov. (type strain R37^T = CECT 7639^T = KCTC 23353^T).     

Aspergillus insulicola sp. nov.

A strain of Aspergillus sp. is described and proposed as a new species under the name ‘Aspergillus insulicola sp. nov.’ Montemayor & Santiago, 1973. This strain was isolated from soil samples taken in ‘Aves Island’ during a scientific expedition. — Aves Island, situated at 15°, 40′, 42″ N and 63°, 36′, 47″ W, about 665 Km of the coast of Venezuela, has very special ecological conditions. Due to its smallness: 550 m long and 40 to 120 m across and to its low profile only 3 m over sea level, it is swept by the sea during the periodical storms and hurricanes in the area. It has thus a very interesting fauna and flora. We took a series of soil samples to study its mycological flora. Forty samples were inoculated by dilution method. In this first paper a species is described and proposed as a new species because of its macroscopic and microscopic characteristics, as well as by its biological properties, under the name ‘Aspergillus insulicola sp. nov.’. In its study we have tried to follow as closely as possible the methods recommended by Kennet B. Raper & Dorothy Fenell, world authorities on the genera Aspergillus and Penicillium. The strain is being kept in USB under the number T1, and has been sent to ATCC & CBSC to be incorporated in their collections.     

Diaminobutyricibacter tongyongensis gen. nov., sp. nov. and Homoserinibacter gongjuensis gen. nov., sp. nov. Belong to the Family Microbacteriaceae

Two bacterial strains, KIS66-7^T and 5GH26-15^T, were isolated from soil samples collected in the South Korean cities of Tongyong and Gongju, respectively. Both strains were aerobic, Gram-stain-positive, mesophilic, flagellated, and rodshaped. A phylogenetic analysis revealed that both strains belonged to the family Microbacteriaceae of the phylum Actinobacteria. The 16S rRNA gene sequence of strain KIS66-7^T had the highest similarities with those of Labedella gwakjiensis KSW2-17^T (97.3%), Cryobacterium psychrophilum DSM 4854T (97.2%), Leifsonia lichenia 2Sb^T (97.2%), Leifsonia naganoensis JCM 10592^T (97.0%), and Cryobacterium mesophilum MSL-15^T (97.0%). Strain 5GH26-15^T showed the highest sequence similarities with Leifsonia psychrotolerans LI1T (97.4%) and Schumannella luteola KHIAT (97.1%). The 16S rRNA gene sequence from KIS66-7^T exhibited 96.4% similarity with that from 5GH26-15^T. Strain KIS66-7^T contained a B2γ type peptidoglycan structure with D-DAB as the diamino acid; MK-13, MK-12, and MK-14 as the respiratory quinones; ai-C_15:0, ai-C_17:0, and i-C_16:0 as the major cellular fatty acids; and diphosphatidylglycerol, phatidylglycerol, and glycolipids as the predominant polar lipids. Strain 5GH26-15T had a B2β type peptidoglycan structure with D-DAB as the diamino acid; MK-14 and MK-13 as the respiratory quinones; ai-C_15:0, i-C_16:0, and ai-C{vn17:0} as the major cellular fatty acids; and diphosphatidylglycerol, phatidylglycerol, and glycolipids as the predominant polar lipids. Both strains had low DNA-DNA hybridization values (<40%) with closely related taxa. Based on our polyphasic taxonomic characterization, we propose that strains KIS66-7^T and 5GH26-15^T represent novel genera and species, for which we propose the names Diaminobutyricibacter tongyongensis gen. nov., sp. nov. (type strain KIS66-7^T=KACC 15515^T=NBRC 108724^T) and Homoserinibacter gongjuensis gen. nov., sp. nov. (type strain 5GH26-15^T=KACC 15524^T=NBRC 108755^T) within the family Microbacteriaceae.     

Proposal of Carbonactinosporaceae fam. nov. within the class Actinomycetia. Reclassification of Streptomyces thermoautotrophicus as Carbonactinospora thermoautotrophica gen. nov., comb. nov

Streptomyces thermoautotrophicus UBT1^T has been suggested to merit generic status due to its phylogenetic placement and distinctive phenotypes among Actinomycetia. To evaluate whether ‘S. thermoautotrophicus’ represents a higher taxonomic rank, ‘S. thermoautotrophicus’ strains UBT1^T and H1 were compared to Actinomycetia using 16S rRNA gene sequences and comparative genome analyses. The UBT1^T and H1 genomes each contain at least two different 16S rRNA sequences, which are closely related to those of Acidothermus cellulolyticus (order Acidothermales). In multigene-based phylogenomic trees, UBT1^T and H1 typically formed a sister group to the Streptosporangiales-Acidothermales clade. The Average Amino Acid Identity, Percentage of Conserved Proteins, and whole-genome Average Nucleotide Identity (Alignment Fraction) values were ≤58.5%, ≤48%, ≤75.5% (0.3) between ‘S. thermoautotrophicus’ and Streptosporangiales members, all below the respective thresholds for delineating genera. The values for genomics comparisons between strains UBT1^T and H1 with Acidothermales, as well as members of the genus Streptomyces, were even lower. A review of the ‘S. thermoautotrophicus’ proteomic profiles and KEGG orthology demonstrated that UBT1^T and H1 present pronounced differences, both tested and predicted, in phenotypic and chemotaxonomic characteristics compared to its sister clades and Streptomyces. The distinct phylogenetic position and the combination of genotypic and phenotypic characteristics justify the proposal of Carbonactinospora gen. nov., with the type species Carbonactinospora thermoautotrophica comb. nov. (type strain UBT1^T, = DSM 100163^T = KCTC 49540^T) belonging to Carbonactinosporaceae fam. nov. within Actinomycetia.     

Three new freshwater species of centrohelid heliozoans: Acanthocystis crescenta sp. nov., A. kirilli sp. nov., and Choanocystis minima sp. nov.

Three new species of centrohelid heliozoans Acanthocystis crescenta, A. kirilli, and Choanocystis minima from two freshwater lakes of Valamo Island (North-Western Russia) were studied using light- and scanning electron microscopy. The main apomorphy of A. kirilli are slightly branched radial scales. Acanthocystis crescenta has characteristic radial scales with crescent-like structures on their tips. Cell diameter of these two species does not exceed 10 μm. Radial scales of C. minima have truncated tips and lack apical teeth; plate scales of this species are oviform, without lateral notches or central rib. The latter organism is the smallest heliozoan species known; the diameter of its body is about 3 μm. The possible domination of small heliozoans in the cryptic diversity of centrohelids is discussed.     

Phylogenetic classification of ten novel species belonging to the genus Bifidobacterium comprising B. phasiani sp. nov., B. pongonis sp. nov., B. saguinibicoloris sp. nov., B. colobi sp. nov., B. simiiventris sp. nov., B. santillanense sp. nov., B. miconis sp. nov., B. amazonense sp. nov., B. pluvialisilvae sp. nov., and B. miconisargentati sp. nov

Ten Bifidobacterium strains, i.e., 6T3, 64T4, 79T10, 80T4, 81T8, 82T1, 82T10, 82T18, 82T24, and 82T25, were isolated from mantled guereza (Colobus guereza), Sumatran orangutan (Pongo abeli), silvery marmoset (Mico argentatus), golden lion tamarin (Leontopithecus rosalia), pied tamarin (Saguinus bicolor), and common pheasant (Phaisanus colchinus). Cells are Gram-positive, non-motile, non-sporulating, facultative anaerobic, and fructose 6-phosphate phosphoketolase-positive. Phylogenetic analyses based on the core genome sequences revealed that isolated strains exhibit close phylogenetic relatedness with Bifidobacterium genus members belonging to the Bifidobacterium bifidum, Bifidobacterium longum, Bifidobacterium pullorum, and Bifidobacterium tissieri phylogenetic groups. Phenotypic characterization and genotyping based on the genome sequences clearly show that these strains are distinct from each of the type strains of the so far recognized Bifidobacterium species. Thus, B. phasiani sp. nov. (6T3 = LMG 32224^T = DSM 112544^T), B. pongonis sp. nov. (64T4 = LMG 32281^T = DSM 112547^T), B. saguinibicoloris sp. nov. (79T10 = LMG 32232^T = DSM 112543^T), B. colobi sp. nov. (80T4 = LMG 32225^T = DSM 112552^T), B. simiiventris sp. nov. (81T8 = LMG 32226^T = DSM 112549^T), B. santillanense sp. nov. (82T1 = LMG 32284^T = DSM 112550^T), B. miconis sp. nov. (82T10 = LMG 32282^T = DSM 112551^T), B. amazonense sp. nov. (82T18 = LMG 32297^T = DSM 112548^T), pluvialisilvae sp. nov. (82T24 = LMG 32229^T = DSM 112545^T), and B. miconisargentati sp. nov. (82T25 = LMG 32283^T = DSM 112546^T) are proposed as novel Bifidobacterium species.     

Minimelanolocus bicolorata sp. nov., Paradendryphiopsis elegans sp. nov. and Corynesporella bannaense sp. nov. from southern China

Minimelanolocus bicolorata sp. nov., Paradendryphiopsis elegans sp. nov. and Corynesporella bannaense sp. nov., are described and illustrated. Minimelanolocus bicolorata is unique in possessing the schizolytic conidial secession and solitary, acropleurogenous, holoblastic conidia that are ellipsoidal, 3-euseptate, 30–35 × 7.5–9.5 μm, verruculose, pale brown, with an appendage at each end. Paradendryphiopsis elegans is characterized by monoblastic conidiogenous cells producing short chains of conidia that are 13–30.5 × 4–6.5 μm, 2–3-euseptate and fusiform to obclavate. Corynesporella bannaense is distinguished by terminal and integrated or discrete, monotretic conidiogenous cells that arise as lateral branches from the tip of the conidiophore and solitary conidia that are 12–16-distoseptate, smooth, obclavate, thick-walled, pale brown, and 100–140 × 10–14 μm. Keys to species of Minimelanolocus, Paradendryphiopsis and Corynesporella are provided.     

Prescottia equi gen. nov., comb. nov.: a new home for an old pathogen

The taxonomic status of Rhodococcus equi, originally isolated from foal specimens, has been the subject of discussion for a number of years. The chequered history of the taxon has prompted this polyphasic analysis of R. equi strains, close members of the genus Rhodococcus and representatives of other genera classified in the order Corynebacteriales, to establish the taxonomic position of this taxon. Thirty one R. equi strains, including the type strain, were examined for genotypic and numerical taxonomic properties. The resultant data are consistent with their classification in the order Corynebacteriales but the R. equi strains formed a distinct phyletic clade away from representatives of other members of the genus Rhodococcus in the 16S rRNA gene tree. Representatives of this clade shared their highest pairwise 16S rRNA gene sequence similarities with the type strain of Rhodococcus kunmingensis (95.2–98.1 %). However, the R. equi taxon was readily distinguished from R. kunmingensis and from the other members of the order Corynebacteriales using a combination of genotypic, chemotypic and phenotypic properties. On the basis of these data the R. equi strains are considered to represent a new genus. The name proposed for this taxon is Prescottia gen. nov., with Prescottia equi comb. nov. as the type species containing the type strain, C 7^T (= ATCC 25729^T = ATCC 6939^T = CCUG 892^T = CIP 54.72^T = DSM 20307^T = HAMBI 2061^T = NBRC 14956^T = JCM 1311^T = JCM 3209^T = LMG 18452^T = NBRC 101255^T = NCTC 1621^T = NRRL B-16538^T = VKM Ac-953^T).     

Fibrophrys columna gen. nov., sp. nov: A member of the family Amphifilidae

A novel Diplophrys-like organism, Fibrophrys columna, was isolated from Hiuchigaike Pond in Japan. F. columna showed a nearly orbicular or broadly elliptical cell shape and has fine filamentous, branching ectoplasmic elements emanating from both polar ends of the cell. Cells also contain orange, amber, or colorless lipid bodies. Although its whole cell morphology resembles that of the genus Diplophrys, Fibrophrys is clearly distinct from Diplophrys on the basis of 18S rDNA sequences. Molecular phylogenetic analysis showed a close relationship of F. columna with Amphifila marina, and its sequence is similar to many environmental stramenopile sequences. The cells of F. columna measured 5.0–8.3 × 5.6–10.3 μm and sometimes possessed hernia-like prongs instead of filamentous ectoplasmic elements. An axis-like electron-dense body was observed in the mitochondria. We also studied the ultrastructure of another Fibrophrys strain, Fibrophrys sp. E-1, which is different from the type strain of F. columna. A ladder-like pattern was recognized in the outer part of unidentified cytoplasmic membranes connected with the mitochondria. The unidentified cytoplasmic membranes were connected to the nuclear, lipid body, and mitochondrial outer membranes. We propose a new genus, Fibrophrys, and a new species, F. columna, based on these ultrastructural and molecular features.     

Paraherbaspirillum soli gen. nov., sp. nov. isolated from soil

A bacterial strain, designated JS5-2^T, was isolated from soil collected from Jeju Island, Republic of Korea. The cells of the strain were Gram-negative, nonspore forming, catalase- and oxidase-positive, aerobic, nonmotile and rod-shaped. Strain JS5-2^T exhibited 96.2–97.2, 95.1–96.3, and 95.4–95.8% 16S rRNA gene sequence similarities to the genera Herbaspirillum, Oxalicibacterium, and Herminiimonas, respectively. The highest sequence similarities were with Herbaspirillum autotrophicum IAM 14942^T (97.2%) and Herbaspirillum frisingense GSF30^T (97.1%). The major fatty acids of strain JS5-2^T were C_16:0 (35.0%), C_17:0 cyclo (19.9%), C_18:1 ω7c (11.4%), and summed feature 3 (C_16:1 ω7c/C_15:0 iso 2-OH) (15.2%), and the major polar lipids of strain JS5-2^T were diphosphatidylglycerol and an unknown aminophospholipid. The strain contained Q-8 as the predominant ubiquinone. DNA-DNA relatedness values between strain JS5-2^T and H. autotrophicum IAM 14942^T, and H. frisingense GSF30^T were 32 and 35%, respectively. The DNA G+C content of strain JS5-2^T was 59.0 mol%. On the basis of phenotypic, genotypic, and physiological evidence, strain JS5-2^T represents a novel species of a new genus, for which the name Paraherbaspirillum soli gen. nov., sp. nov. is proposed. The type strain JS5-2^T (=KACC 12633^T =NBRC 106496^T) is proposed.     

Position stratigraphique de Pseudolituonella? guerneti nov. sp. en Eubee (Grece)

The stratigraphic position of Pseudolituonella? guerneti n. sp. in the Asprochroma section of Eubee (Greece, pelagonian zone) is discussed.     

Natronogracilivirga saccharolytica gen. nov., sp. nov. and Cyclonatronum proteinivorum gen. nov., sp. nov., haloalkaliphilic organotrophic bacteroidetes from hypersaline soda lakes forming a new family Cyclonatronaceae fam. nov. in the order Balneolales

Two heterotrophic bacteroidetes strains were isolated as satellites from autotrophic enrichments inoculated with samples from hypersaline soda lakes in southwestern Siberia. Strain Z-1702^T is an obligate anaerobic fermentative saccharolytic bacterium from an iron-reducing enrichment culture, while Ca. Cyclonatronum proteinivorum Omega^T is an obligate aerobic proteolytic microorganism from a cyanobacterial enrichment. Cells of isolated bacteria are characterized by highly variable morphology. Both strains are chloride-independent moderate salt-tolerant obligate alkaliphiles and mesophiles. Strain Z-1702^T ferments glucose, maltose, fructose, mannose, sorbose, galactose, cellobiose, N-acetyl-glucosamine and alpha-glucans, including starch, glycogen, dextrin, and pullulan. Strain Omega^T is strictly proteolytic utilizing a range of proteins and peptones. The main polar lipid fatty acid in both strains is iso-C_15:0, while other major components are various C₁₆ and C₁₇ isomers. According to pairwise sequence alignments using BLAST Gracilimonas was the nearest cultured relative to both strains (<90% of 16S rRNA gene sequence identity). Phylogenetic analysis placed strain Z-1702^T and strain Omega^T as two different genera in a deep-branching clade of the new family level within the order Balneolales with genus. Based on physiological characteristics and phylogenetic position of strain Z-1702^T it was proposed to represent a novel genus and species Natronogracilivirga saccharolityca gen. nov., sp. nov. (= DSMZ 109061^T =JCM 32930^T =VKM B 3262^T). Furthermore, phylogenetic and phenotypic parameters of N. saccharolityca and C. proteinivorum gen. nov., sp. nov., strain Omega^T (=JCM 31662^T, =UNIQEM U979^T), make it possible to include them into a new family with a proposed designation Cyclonatronaceae fam. nov..     

Heterogeneity of the yeasts Zygowilliopsis californica: Z. californica var. dimennae comb. nov., stat. nov. and Z. californica var. fukushimae comb. nov., stat. nov.

A significant heterogeneity of the species Zygowilliopsis californica was revealed using RFLP-analysis of the PCR-amplified rDNA fragment spanning the 5.8S rRNA gene and the internal transcribed spacers ITS1 and ITS2. Phylogenetic analysis of the nucleotide sequences of ITS1 and ITS2 rDNA differentiated three varieties: Z. californica var. californica, Z. californica var. dimennae, and Z. californica var. fukushimae. The most variable was the ITS2 region, where 7–26 nucleotide substitutions were revealed. The varieties formed semisterile hybrids with meiotic segregation of control markers. The limits of the phylogenetic species concept are discussed.     

Syntrophomonas wolfei gen. nov. sp. nov., an Anaerobic, Syntrophic, Fatty Acid-Oxidizing Bacterium

An anaerobic, nonphototrophic bacterium that β-oxidizes saturated fatty acids (butyrate through octanoate) to acetate or acetate and propionate using protons as the electron acceptor (H₂ as electron sink product) was isolated in coculture with either a non-fatty acid-degrading, H₂-utilizing Desulfovibrio sp. or methanogens. Three strains of the bacterium were characterized and are described as a new genus and species, Syntrophomonas wolfei. S. wolfei is a gram-negative, slightly helical rod with round ends that possesses between two to eight flagella laterally inserted along the concave side of the cell. It has a multilayered cell wall of the gram-negative type. The presence of muramic acid, inhibition of growth by penicillin, and increased sensitivity of the cells to lysis after treatment with lysozyme indicate that peptidoglycan is present in the cell wall. Cells of S. wolfei contain poly-β-hydroxybutyrate. Isoheptanoate was degraded to acetate, isovalerate, and H₂. Carbohydrates, proteinaceous materials, alcohols, or other tested organic compounds do not support growth. Common electron acceptors are not utilized with butyrate as the electron donor. Growth and degradation of fatty acids occur only in syntrophic association with H₂-using bacteria. The most rapid generation time obtained by cocultures of S. wolfei with Desulfovibrio and Methanospirillum hungatei is 54 and 84 h, respectively. The addition of Casamino Acids but neither Trypticase nor yeast extract stimulated growth and resulted in a slight decrease in the generation time of S. wolfei cocultured with M. hungatei. The addition of H₂ to the medium stopped growth and butyrate degradation by S. wolfei.     

Lacipirellula parvula gen. nov., sp. nov., representing a lineage of planctomycetes widespread in low-oxygen habitats,description of the family Lacipirellulaceae fam. nov. and proposal of the orders Pirellulales ord. nov., Gemmatales ord. nov. and Isosphaerales ord. nov.

Pirellula-like planctomycetes are ubiquitous aquatic bacteria, which are often detected in anoxic or micro-oxic habitats. By contrast, the taxonomically described representatives of these bacteria, with very few exceptions, are strict aerobes. Here, we report the isolation and characterization of the facultatively anaerobic planctomycete, strain PX69^T, which was isolated from a boreal lake. Its 16S rRNA gene sequence is affiliated with the Pirellula-related Pir4 clade, which is dominated by environmental sequences retrieved from a variety of low-oxygen habitats. Strain PX69^T was represented by ellipsoidal cells that multiplied by budding and grew on sugars, some polysaccharides and glycerol. Anaerobic growth occurred by means of fermentation. Strain PX69^T grew at pH 5.5–7.5 and at temperatures between 10 and 30 °C. The major fatty acids were C18:1ω9c, C16:0 and C16:1ω7c; the major intact polar lipid was dimethylphosphatidylethanolamine. The complete genome of strain PX69^T was 6.92 Mb in size; DNA G + C content was 61.7 mol%. Among characterized planctomycetes, the highest 16S rRNA gene similarity (90.4%) was observed with ‘Bythopirellula goksoyri’ Pr1d, a planctomycete from deep-sea sediments. We propose to classify PX69^T as a novel genus and species, Lacipirellula parvula gen. nov., sp. nov.; the type strain is strain PX69^T (=KCTC 72398^T = CECT 9826^T = VKM B-3335^T). This genus is placed in a novel family, Lacipirellulaceae fam. nov., which belongs to the order Pirellulales ord. nov. Based on the results of comparative genome analysis, we also suggest establishment of the orders Gemmatales ord. nov. and Isosphaerales ord. nov. as well as an emendation of the order Planctomycetales.     

Description ofOceanospirillum kriegii sp. nov. andO. jannaschii sp. nov. and assignment of two species ofAlteromonas to this genus asO. commune comb. nov. andO. vagum comb. nov

Immunological studies of Fe-containing superoxide dismutase (FeSOD) and glutamine synthetase (GS) have established a close relationship betweenOceanospirillum linum (the type species of the genus),O. beijerinckii, Alteromonas communis, A. vaga, and two unnamed species of marine bacteria (groups H-1 and I-1). The four latter species have, consequently, been assigned to the genusOceanospirillum asO. commune comb. nov.,O. vagum comb. nov.,O. kriegii sp. nov. (group H-1; type strain 197, ATCC 27133), andO. jannaschii sp. nov. (group I-1; type strain 207, ATCC 27135). The phenotypic properties of these species are presented together with their distinguishing traits.     

Proposal of Ensifer psoraleae sp. nov., Ensifer sesbaniae sp. nov., Ensifer morelense comb. nov. and Ensifer americanum comb. nov

In a survey of rhizobia associated with the native legumes in Yunnan Province, China, seven and nine strains isolated from the root nodules of Psoralea corylifolia, Sesbania cannabina and Medicago lupulina were respectively classified into the novel genomic species groups I and II in the genus Ensifer (former Sinorhizobium) based on the sequence analyses of the 16S rRNA gene. Analyses of concatenated housekeeping genes (atpD, recA and glnII) further revealed that they were distinct lineages in the genus, and group I was most similar to Ensifer terangae and Ensifer garamanticus (both with 94.2% similarity), while group II was most similar to Ensifer adhaerens (94.0%). These groups could be distinguished from closely related species by DNA–DNA relatedness, MALID-TOF MS, cellular fatty acid profiles and a series of phenotypic characters. Therefore, two novel species were proposed: Ensifer psoraleae sp. nov. (seven strains, type strain CCBAU 65732^T = LMG 26835^T = HAMBI 3286^T) and Ensifer sesbaniae sp. nov. (nine strains, type strain CCBAU 65729^T = LMG 26833^T = HAMBI 3287^T). They had a DNA G + C mol% (T_m) of 58.9 and 60.4, respectively. Both of the type strains formed effective nodules on common bean (Phaseolus vulgaris) and their hosts of origin. In addition, the previously described species Sinorhizobium morelense and Sinorhizobium americanum were renamed as Ensifer morelense comb. nov. and Ensifer americanum comb. nov. according to the accumulated data from different studies.