The distribution of Oligochaeta on an exposed rocky shore in southeast Ireland

The distribution of oligochaetes was studied on a wave-exposed, granite shore at Carnsore Point, County Wexford. Habitats sampled were crevices and shallow surface cracks in rock, Lichina pygmaea turf, a Mytilus edulis bed, barnacles, Laurencia pennatifida turf, Corallina officinalis turf and pools. Lichina, mussels, barnacles greater than 8 mm in height, Laurencia and Corallina were sampled quantitatively.Three species of Tubificidae and nine species of Enchytraeidae were recorded including two new species described from this locality and five which are undescribed. Oligochaetes were present in all the habitats sampled but were rare in pools and numbers were low on barnacle-covered rock except where barnacles were more than 8 mm in height. The number of species increased with decreasing tide level and was highest in lower shore crevices. Highest densities were recorded from Corallina turf and in barnacles > 8 mm. Lumbricillus semifuscus was the most widespread species, occurring in crevices and mats at all tide levels; other species had more restricted distributions. Grania species were confined to Corallina turf. Only three species of oligochaete were recorded from cracks where they were almost the only fauna present. Species diversity and density were influenced by physical structure of the habitat, particularly the amount of retained sediment.The oligochaetes are members of a rich cryptofauna in habitats which provide them with organic matter and moisture and protect them from environmental extremes and wave damage and from predators during immersion. Reasons for the scarcity of oligochaete records from exposed rocky shores and the high proportion of new species in this study are discussed.     

Soil mycoflora from trufflè native areas of Saudi Arabia

Soil fungi of areas in the North-Eastern region of Saudi Arabia where truffles are native were surveyed. Forty-three species of fungi belonging to twenty genera were isolated. Most were recovered from soils underneath or around truffle ascocarps: thirty species from soil under the surface of Tirmania nivea ascocarps, twenty-four from Terfezia boudieri soil and twenty species each from Tirmania pinoyi and Terfezia claveryi soils. Rhizosphere soil of Helianthemum lippi, on the other hand, yielded twenty-four fungal species while only fourteen fungal species were found in soil without vegetation. The total counts of fungi/g soil were highest in soils from the under surface of truffles, followed by rhizosphere soil, with the lowest in soils without vegetation.     

Assessment of Water Pollution using Diatom Community Structure and Species Distribution — A Case Study in a Tropical River Basin

A study on biological assessment of water pollution using diatom community structure and species distribution was carried out in the Linggi River Basin, Malaysia which was polluted by various urban, industrial and agricultural wastes. A total of 86 diatom taxa belonging to 21 genera were recorded from all eight sampling stations located in the basin, of which 70 species were found on artificial substrates; the remaining 16 species were recorded exclusively on natural substrates. The number of diatom species observed between the stations varied from 22-47 species. The dominant diatom species in decreasing order of abundance were Eunotia vanheurckii, Nitzschia palea, Achnanthes saxonica, Gomphonema parvulum and Achnanthes minutissima. The most common clean water species were Achnanthes minutissima, A. linearis and Synedra rumpens. The most tolerant species were Nitzschia palea followed by Gomphonema parvulum and Pinnularia braunii. Eunotia vanheurckii and Navicula cryptocephala occurred in high densities at both unpolluted and polluted stations and can be considered as the common facultative or indifferent species. Although a large number of species were recorded at the unpolluted stations, equivalent number of species were also found at the moderately polluted stations. However, the number of species was reduced at grossly polluted stations. Nevertheless, a marked variation in species association exists between the unpolluted and polluted stations, but not among the polluted stations to distinguish the type and degree of pollution.     

Diversity and Coexistence of Ectoparasites in Small Rodents in a Tropical Dry Forest

To understand the mechanisms driving species diversity is central to community ecology. Here, we explored if habitat partitioning is associated with a species‐rich ectoparasite community in small rodents from a tropical dry forest in western Mexico. We trapped 199 mice in three 0.5 ha‐plots from eight small rodent species for every two months, from July 2011 to April 2012, and collected their ectoparasites. We identified 17 species of mites, two sucking lice species, two phoretic species, and one commensal species. The most abundant ectoparasite species was Steptolaelaps liomydis, representing 42 percent of all ectoparasites collected; seven ectoparasite species had < 10 individuals. Eighteen ectoparasite species (of 22 species) were collected from the most abundant rodent Liomys pictus. C‐score and the number of checkerboard species pairs were significantly higher against a random expectation. Ectoparasite species in L. pictus mice showed host microhabitat partitioning; Fahrenholzia ehrlichi and Fahrenholzia texana were found only in the anterior dorsal area, Ornithonysus sp. occurred along the dorsal part, Ixodes species were restricted to the ears, and Steptolaelaps liomydis was found throughout the body. We also identified ectoparasite communities with distinct species composition in two rodent species that use contrasting macrohabitats (L. pictus, strictly terrestrial; Peromyscus perfulvus, mostly arboreal). The remaining and low abundant rodent species showed a species‐poor ectoparasite community composition. We conclude that habitat partitioning at both macro and microhabitat scales appeared to characterize the species‐rich ectoparasite community. Conversely, most rodent host species with low abundances showed a species‐poor ectoparasite community.     

Development of novel microsatellite markers for the grassland species Lolium multiflorum,Lolium perenne and Festuca pratensis

Twelve microsatellite markers were isolated from Lolium multiflorum. Allelic variability and cross‐species amplification were assessed on 16 individuals of each of the three grassland species L. multiflorum, Lolium perenne and Festuca pratensis. Cross‐species amplification success was 100% for L. perenne and 83% for F. pratensis. The number of alleles detected ranged from one to 14 with an average of 3.4. While three microsatellite loci were polymorphic in all three species, one marker produced species‐specific alleles in all three species. These microsatellite markers provide a valuable tool for population genetic studies within and among species of the Festuca–Lolium complex.     

LYCOSPORA FROM CARBONIFEROUS LEPIDOSTROBUS COMPRESSIONS

Spores were extracted from Carboniferous Lepidostrobus compressions in order to associate in situ microspores with dispersed species of Lycospora. Two hundred twenty-six cones were examined, of which 61 contained spores. Fertile cones came from the Westphalian D of England, Namurian B through Westphalian D of the Appalachian and Illinois basins, and the Westphalian D of the Western Interior. Cones were separated into species based on microspore and cone morphology. Lycospora trigonoreticulata was produced by Lepidostrobus princeps from Westphalian C-D rocks from Missouri, the Illinois Basin, and the Appalachian Basin. Lycospora rotunda was produced by Lepidostrobus sp. A from Westphalian A rocks of Alabama. Two cone species produced Lycospora torquifer: Lepidostrobus praelongus from the Westphalian D of Pennsylvania and Lepidostrobus variabilis from the Westphalian A and C of the Illinois and Appalachian basins. Lycospora punctata was produced by Lepidostrobus cf. squarrosus from the Westphalian D of England, the Appalachian Basin, and Illinois Basin. Lycospora noctuina was produced by Lepidostrobus haslingdenensis from the Namurian B/C of Illinois. Microspore species are differentiated primarily on the basis of size, cingulum structure and width, and ornamentation. Cone species differ in width and distal lamina size, shape, and attitude. Lycospora species isolated from clastic species of Lepidostrobus differ completely from those of coal-swamp species, confirming that lycopod trees from clastic environments represent biologically different species from those centered in coal swamps.     

基于可培养方法分析云南腾冲小空山火山谷芽胞杆菌分布特征

【目的】为了解云南腾冲小空山火山谷土壤中可培养芽胞杆菌种类分布特征。【方法】采用可培养手段对小空山火山谷阳坡、谷底和阴坡土壤中的芽胞杆菌进行分离培养,根据16S rRNA基因序列同源性对分离菌株进行鉴定,并分析系统发育地位。利用Canoco5软件分析采样点芽胞杆菌种类分布特征与土壤样品理化性质的相关性。【结果】从火山谷土壤样品中共分离获得180株芽胞杆菌,16S rRNA基因测序鉴定结果表明分离菌株隶属于芽胞杆菌纲2个科(芽胞杆菌科和类芽胞杆菌科)、6个属、34个种,其中芽胞杆菌属(Bacillus) 11个种,类芽胞杆菌属(Paenibacillus) 14个种,短芽胞杆菌属(Brevibacillus)3个种,赖氨酸芽胞杆菌属(Lysinibacillus)4个种,嗜冷芽胞杆菌属(Psychrobacillus)1个种和绿芽胞杆菌属(Viridibacillus)2个种,其中7个菌株与其最近模式菌株16SrRNA相似性低于种的界定阈值(98.65%),为芽胞杆菌潜在新物种。优势属为芽胞杆菌属和类芽胞杆菌属,优势种为蕈状芽胞杆菌(Bacillusmycoides),图瓦永芽胞杆菌(Bacillustoyonensis),蜡状芽胞杆菌(Bacilluscereus),解木糖赖氨酸芽胞杆菌(Lysinibacillusxylanilyticus),蜂房类芽胞杆菌(Paenibacillusalvei)和沙地绿芽胞杆菌(Viridibacillus arenosi)。其中16个种分离自阳坡,29个种分离自阴坡,9个种分离自谷底,三者共同种类为6种。阳坡、谷底和阴坡的芽胞杆菌种群分布Bray-Curtis相似性为62.4%,多样性分析结果表明,Shannon指数(H′)大小次序为阴坡阳坡谷底。环境因子分析发现,芽胞杆菌种群分布多样性特征与其土壤的海拔高度、碳氮比和硫含量呈负相关,而和碳源和氮源含量呈正相关。【结论】从以上结果得出,云南腾冲火山谷有着较为丰富的芽胞杆菌资源,且还存在可分离培养的芽胞杆菌的潜在新物种,为利用火山微生物资源提供了保障。     

Best linear unbiased prediction of host-range of the facultative parasite Colletotrichum gloeosporioides f. sp. salsolae, a potential biological control agent of Russian thistle

Russian thistle or tumbleweed (Salsola tragus L.) is an introduced invasive weed in N. America. It is widely distributed in the US and is a target of biological control efforts.The fungus Colletotrichum gloeosporioides (Penz.) Penz. & Sacc. in Penz. f. sp. salsolae (CGS) is a facultative parasite under evaluation for classical biological control of this weed. Host-range tests were conducted with CGS in quarantine to determine whether the fungus is safe to release in N. America. Ninetytwo accessions were analyzed from 19 families: Aizoaceae, Alliaceae, Amaranthaceae, Apiaceae, Asteraceae, Brassicaceae, Cactaceae, Campanulaceae, Chenopodiaceae, Cucurbitaceae, Cupressaceae, Fabaceae, Malvaceae, Nyctaginaceae, Phytolaccaceae, Poaceae, Polygonaceae, Sarcobataceae, and Solanaceae and 10 tribes within the Chenopodiaceae: Atripliceae, Beteae, Camphorosmeae, Chenopodieae, Corispermeae, Halopepideae, Polycnemeae, Salicornieae, Salsoleae, and Suaedeae. These included 62 genera and 120 species. To facilitate interpretation of results, disease reaction data were combined with a relationship matrix derived from internal transcribed spacer DNA sequences and analyzed with mixed model equations to produce Best Linear Unbiased Predictors (BLUPs) for each species. Twenty-nine species (30 accessions) from seven closely-related Chenopodiaceae tribes had significant levels of disease severity as indicated by BLUPs, compared to six species determined to be susceptible with least squares means estimates. The 29 susceptible species were: 1 from Atripliceae, 4 from Camphorosmeae, 1 from Halopepideae, 2 from Polycnemeae, 6 from Salicornieae, 8 from Salsolae, and 7 from Suaedeae. Most species in the genus Salsola, which are all introduced and weedy, were very susceptible and damaged by CGS. Statistical comparisons and contrasts of BLUPs indicated that these Salsola species were significantly more susceptible than non-target species, including 15 species from relatives in the closely-related genera Bassia (=Kochia), Nitrophila, Salicornia, Sarcocornia, and Suaeda. Of the 29 susceptible species, 10 native or commercially important species in N. America were identified as needing additional tests to determine the extent of any damage caused by infection.     

Incidence of dermatophytes and cyclohexamide resistantfungi on healthy children hairs and nails in nurseries

In order to estimate the prevalence of dermatophytes and other fungi on healthy children hairs and nails, 92 hair samples and 85 nail samples (groups of 10 finger nails from each child) were collected from 5 nurseries (children aged 9 months up to 4 years) in Assiut city. From hair samples 22 species were collected, Trichophyton (2 species) and Microsporum (2 species) were the only recovered dermatophytes in addition to well known keratinophilic genus Chrysosporium (4 species). From nail samples, 18 species were identified, Trichophyton was represented by 4 species, Microsporum, 2 species and Chrysosporium, 4 species. Also, several other saprophytes and cycloheximide resistant fungi were isolated. This revised version was published online in June 2006 with corrections to the Cover Date.     

Niche segregation among sympatric Amazonian teiid lizards

We examined standard niche axes (time, place, and food) for three sympatric teiid lizards in the Amazon rain forest. Activity times during the day were similar among species. Ameiva ameiva were in more open microhabitats and had higher body temperatures compared with the two species of Kentropyx. Microhabitat overlaps were low and not significantly different from simulations based on Monte Carlo analysis. Grasshoppers, crickets, and spiders were important in the diets of all three species and many relatively abundant prey were infrequently eaten (e.g., ants). Dietary overlaps were most similar between the two species of Kentropyx even though microhabitat overlaps were relatively low. A Monte Carlo analysis on prey types revealed that dietary overlaps were higher at all ranks than simulated overlaps indicating that use of prey is not random. Although prey size was correlated with lizard body size, there were no species differences in adjusted prey size. A. ameiva ate more prey items at a given body size than either species of Kentropyx. Body size varies among species, with A. ameiva being the largest and K. altamazonica the smallest. The two species of Kentropyx are most distant morphologically, with A. ameiva intermediate. The most distant species morphologically are the most similar in terms of prey types. A morphological analysis including 15 species from four genera revealed patterns of covariation that reflected phylogenetic affinities (i.e., taxonomic patterns are evident). A cluster analysis revealed that A. ameiva, K. pelviceps, and K. altamazonica were in the same morphological group and that within that group, A. ameiva differed from the rest of the species. In addition, K. pelviceps and K. altamazonica were distinguishable from other species of Kentropyx based on morphology. Received: 26 December 1998 / Accepted: 15 September 1999     

Food selection by freshwater snails in the Gezira irrigation canals,Sudan

Stomach content analysis was carried out on samples of the freshwater snail species Biomphalaria pfeifferi, Bulinus truncatus, Bulinus forskalii (Pulmonata, Planorbidae), Lymnaea natalensis (Pulmonata, Lymnaeidae), Melanoides tuberculata, Cleopatra bulimoides (Prosobranchia, Thiaridae) and Lanistes carinatus (Prosobranchia, Ampullariidae) from different irrigation canals in Sudan. In order to evaluate overlap in diet selection among these species, sites with two or more of the above-mentioned species present were selected. For some species food choice was examined in relation to size groupings. In addition, samples of Marisa cornuarietis (Prosobranchia, Ampullariidae) from small ponds in Sudan, samples of Biomphalaria pfeifferi and Helisoma duryi (Pulmonata, Planorbidae) from drainage canals in an irrigation scheme in northern Tanzania, and samples of H. duryi from fish ponds in the coastal area of Kenya were also analysed. The results indicate a great similarity in the food choice of these species, especially among the pulmonate species. All species feed on fine detritus, epiphytic algae and decaying macrophytes. No fresh fragments of aquatic macrophytes were found and animal remains were found only on a few occasions. However, the stomach contents of the ampullarid species were characterized by large fragments of dead macrophyte tissue, while the composition of the finer particles showed a great resemblance to that of the pulmonate species. The diet of the thiarid species is essentially the same as that of the pulmonate species, although in one site Cleopatra bulimoides showed a greater preference for green algae. Apart from the avoidance of blue-green algae, there was little evidence of selection of certain algal components of the Aufwuchs for the pulmonate species. Detritus constitutes the major component of the stomach content of all these snail species.     

中国新记录种黑毛地蜂记述（膜翅目：地花蜂科：毛地蜂亚科）

对近来记述的古北区东部地区的毛地蜂属Panurginus Nylander全部14个种作了补充研究。中国对该属的研究较少,仅知2种：黑足毛地蜂Panurginus nigripes Morawitz 和黄跗毛地蜂Panurginus flavotarsus ? Wu。黑毛地蜂Panurginus niger Nylander为该属的模式种,分布于俄罗斯和蒙古,本文报道该种为中国新记录种。目前,中国已知该属3种,分属于2个组即niger-group和herzi-group。文中提供了中国已知种类检索表。     

Molecular phylogeny of black flies (Diptera: Simuliidae) from Thailand, using ITS2 rDNA

The sequences of the second internal transcribed spacer (ITS2) of ribosomal DNA (rDNA) were determined for 40 black fly species from Thailand, belonging to 4 subgenera of the genus Simulium, namely Gomphostilbia (12 species), Nevermannia (5 species), Montisimulium (1 species), Simulium sensu stricto (21 species), and an unknown subgenus with one species (Simulium baimaii). The length of the ITS2 ranged from 247 to 308 bp. All black fly species had high AT content, ranging from 71 to 83.8%. Intraindividual variation (clonal variation) occurred in 13 species, ranging from 0.3 to 1.1%. Large intrapopulation and interpopulation heterogeneities exist in S. feuerboni from the same and different locations in Doi Inthanon National Park, northern Thailand. Phylogenetic relationships among 40 black fly species were examined using PAUP (version 4.0b10) and MrBAYS (version 3.0B4). The topology of the trees revealed two major monophyletic clades. The subgenus Simulium and Simulium baimaii were placed in the first monophyletic clade, whereas the subgenera Nevermannia + Montisimulium were placed as the sister group to the subgenus Gomphostilbia in the second monophyletic clade. Our results suggest that S. baimaii belongs to the malyschevi-group or variegatum-group in the subgenus Simulium. The molecular phylogeny generally agrees with existing morphology-based phylogenies.     

Influence of mineral nutrient availability on growth of tree seedlings from the tropical deciduous forest

The effects of different nutrient availabilities on growth and biomass partitioning in seedlings from the tropical deciduous forest in Mexico were compared. The tree species studied were Heliocarpus pallidus, a species associated with disturbed parts of the forest, and Caesalpinia eriotachys, Jacquinia pungens and Recchia mexicana, species from mature, undisturbed habitats. The tropical deciduous tree seedlings were grown in pure silica sand for 50 days inside growth chambers under four nutrient regimes; 5, 20, 100 and 200% Long Ashton nutrient solutions. Data showed contrasting responses among species to different nutrient availabilities. Except Jacquinia pungens, all species had increased growth and productivity as nutrient level increased from 5 to 100%; however, no significant differences in these parameters were detected between 100 and 200% in all species. Compared with mature forest species, pioneer species showed higher variations in biomass production, relative growth rate and net assimilation rate. In contrast to mature forest species, root/ shoot ratios in Heliocarpus pallidus were greater and thus showed higher biomass allocation to roots when nutrient supply was limited. This response suggests higher phenotypic plasticity in pioneer species. Species from mature parts of the forest (Caesalpinia eriostachys, Recchia mexicana) showed less dependency on nutrient supply than pioneer species. These responses appear to support observations from studies with temperate plants investigating growth responses to soil fertility.     

Capacity of Japanese Asobara species (Hymenoptera; Braconidae) to parasitize a fruit pest Drosophila suzukii (Diptera; Drosophilidae)

Drosophila suzukii is a pest of soft fruit such as cherry, strawberry or blueberry and recently colonized North America and Europe from Asia. In this study, we assessed the utility of Japanese Asobara species as agents for the biological control of this pest species by examining their host use in nature and their capacity to parasitize this pest species. From Japan, eight Asobara species including putative three undescribed species were found; Asobara japonica was recorded from all over Japan, Asobara tabida from high altitude areas in central and northern Japan, Asobara rossica and Asobara rufescens from central to northern Japan, three undescribed species from central to western or southern Japan and Asobara pleuralis from a subtropical island of Japan. Among them, an undescribed species Asobara sp. TK1 may be useful as an agent for the biological control because it was recorded only from D. suzukii. All of the remaining seven species were generalists mainly using drosophilid species associated with fermenting fruits, and four of them had no or very low abilities to parasitize D. suzukii, indicating that these seven species are not or less appropriate as agents for biological control.     

Marine mycoflora of south India with special emphasis on lignicolous marine fungi

A study dealing with the marine fungi associated with decaying wood samples in the brackish water mangrove ecosystem and shoreline ecosystem was carried out in south India. A total of 19 marine fungi were isolated from the brackish water mangrove ecosystem. They included 13 Ascomycetes, one Basidiomycete and five Mitosporic fungi. In terms of percent frequency of occurrence, the most frequent species obtained from the brackishwater were the Lignincola longirostris (16.60%) and Savoryella lignicola (12.09%). Nine species were found frequently. Five species were occasionally encountered. Aigialus mangrovei, Aniptodera mangrovei and Halosarpheia marina were the rare species recorded. The average number of isolates per wood sample was 1.53. A total of 27 marine fungi including 15 ascomycetes, one basidiomycete and ten mitosporic fungi were recorded from the shoreline ecosystem. In terms of percent frequency of occurrence, the most frequent species obtained from Kanyakumari were the Arenariomyces trifurcates (13.66%), Corollospora maritima (12.44%), and Cirrenalia pygmea (10.98%). Seven species were found frequently. Fourteen species were occasionally encountered. Three species were found to be rare in occurrence. The average number of isolates per wood sample was 1.21.     

Isolation for bacteria and fungi for the hydrolysis of phthalate and terephthalate esters

Bacterial and fungal strains were isolated from enrichment cultures using diethylphthalate, diethylterephthalate, or ethylene glycol dibenzoate as sole carbon sources.Aureobacterium, Flavobacterium, andMicrococcus species were isolated from diethylphthalate enrichments;Rhodococcus andXanthomonas species were isolated from diethylterephthalate enrichments;Rhodococcus andFusarium species were isolated from ethylene glycol dibenzoate enrichments.     

Keratinophilic fungi of poultry farm and feather dumping soil in Tamil Nadu,India

Soils of 10 poultry farms from Namakkal and 12 feather dumping sites from Chennai were studied for the presence of keratinophilic fungi. A total of 34 species belonging to 19 genera and one non-sporulating fungus were recovered. Sixteen species of fungi and one non-sporulating fungi were common to both sites, eight species were specific to Namakkal and nine species were specific to Chennai. Dermatophytes and closely related fungi were represented by six species belonging to five genera. Fungal species commonly found in the soil samples included Chrysosporium keratinophilum (73%), Trichophyton mentagrophytes (68.2%), Microsporum gypseum (64%), Myceliopthora vellerea (32%), Chrysosporium state of Arthroderma tuberculatum (27.3%) and Geomyces pannorum (23%). Non-dermatophyte fungi were represented by 28 species belonging to 14 genera and one non-sporulating fungus.     

A study of fungal spore poupulations in the atmosphere of Kuwait

The fungal air-spora of Kuwait was investigated using the Petri-dish trapping technique. During the period from 1st April, 1974 to 30th June, 1975, a total of 3685 colonies were recorded from 2 % malt agar plates. Fifty-five genera and 116 species were identified. Alternaria occupied the first place in the order of percentage incidence, being represented by 18.3% of the entire catch, followed by Aspergillus (17.1%), Penicillium (14%), Cladosporium (13.6%), Drechslera (13.3%), and Ulocladium (7.1%). The widest spectrum of species was displayed by Aspergillus (possessed 19 species) followed by Penicillium (17 species), Alternaria (6 species), Drechslera (5 species), and Ulocladium (4 species). The monthly total number of fungi showed marked seasonal periodicity with the greatest number of colonies occurring in March–April and November. The lowest counts were recorded in mid-summer (July) and mid-winter (January). Only 220 colonies and 17 species of thermophilous fungi were reported from plates incubated at 45 °C. The most common species were Aspergillus fumigatus and A. terreus.     

A review of the West Palaearctic aphidiines (Hymenoptera: Braconidae: Aphidiinae) parasitic on Uroleucon spp., with the description of a new species

A review of aphidiine wasps (Hymenoptera: Braconidae) parasitizing the Uroleucon species in the West Palaearctic is presented. Eleven species are keyed and illustrated. In addition, a new hymenopteran parasitoid species: Praon nonveilleri n. sp. from Uroleucon inulicola (Hille Ris Lambers) infesting Inula ensifolia L., is described. The new species is diagnosed and illustrated. It belongs to the “dorsale-yomenaé” species group and was collected from the Djetinja canyon in Serbia and Montenegro. The aphidiines presented in this work were identified from 97 aphid taxons occurring on 236 plant taxons. Furthermore, 361 original parasitoid – host aphid – host plant associations of the species mentioned in the key are presented. Finally, phylogenetic relationships inside the “dorsale-yomenae” species group and related species were reconstructed using cladistic distance methods.