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1.
Environmental DNA (eDNA) analysis has recently been used as a new tool for estimating intraspecific diversity. However, whether known haplotypes contained in a sample can be detected correctly using eDNA‐based methods has been examined only by an aquarium experiment. Here, we tested whether the haplotypes of Ayu fish (Plecoglossus altivelis altivelis) detected in a capture survey could also be detected from an eDNA sample derived from the field that contained various haplotypes with low concentrations and foreign substances. A water sample and Ayu specimens collected from a river on the same day were analysed by eDNA analysis and Sanger sequencing, respectively. The 10 L water sample was divided into 20 filters for each of which 15 PCR replications were performed. After high‐throughput sequencing, denoising was performed using two of the most widely used denoising packages, unoise3 and dada2 . Of the 42 haplotypes obtained from the Sanger sequencing of 96 specimens, 38 (unoise3 ) and 41 (dada2 ) haplotypes were detected by eDNA analysis. When dada2 was used, except for one haplotype, haplotypes owned by at least two specimens were detected from all the filter replications. Accordingly, although it is important to note that eDNA‐based method has some limitations and some risk of false positive and false negative, this study showed that the eDNA analysis for evaluating intraspecific genetic diversity provides comparable results for large‐scale capture‐based conventional methods. Our results suggest that eDNA‐based methods could become a more efficient survey method for investigating intraspecific genetic diversity in the field.  相似文献   

2.
A cell-counting algorithm, developed in Matlab®, was created to efficiently count migrated fluorescently-stained cells on membranes from migration assays. At each concentration of cells used (10,000, and 100,000 cells), images were acquired at 2.5 ×, 5 ×, and 10 × objective magnifications. Automated cell counts strongly correlated to manual counts (r2 = 0.99, P < 0.0001 for a total of 47 images), with no difference in the measurements between methods under all conditions. We conclude that our automated method is accurate, more efficient, and void of variability and potential observer bias normally associated with manual counting.  相似文献   

3.
Perithecia of Erysiphe cichoracearum (powdery mildew) were found on several, but not all, accessions of Lactuca serriola, L. saligna, L. aculeata and L. serriola × L. sativa hybrid. Their occurrence varied between Lactuca species and different accessions of the same species. Perithecia were globular, had a mean diameter of 125.5 μm and contained an average 13–14 asci. Asci had mean dimensions of 55.0 × 26.1 μm and each contained two ascospores which measured an average 17.4 × 10.6 μm. There were however significant differences in the dimensions of spores and other structures obtained from different Lactuca accessions.  相似文献   

4.
Scanning electron microscopy has become an increasingly useful tool for anthropologists, particularly because of the development of improved methods of replicating specimens. One of the best replication techniques involves silicone-based dental impression materials to make negative impressions, in conjunction with epoxy resins, which are used to make positives or casts. The technique outlined here is particularly useful for anthropologists. Using this technique allows the examination of bone surfaces, teeth, and fossils for taphonomic, microwear, and experimental studies. Reproduction of detail is faithful at magnifications of × 1,500 to × 2,000, routinely giving resolutions of .1 to .25 μm.  相似文献   

5.
doi:10.1111/j.1741‐2358.2009.00332.x
Effect of thermal cycling on microleakage between hard chairside relines and denture base acrylic resins Objectives: Microleakage is a pre‐stage of debonding between hard chairside relines and denture base acrylic resins. Therefore, it is important to assess them with regard to the longevity of the relined denture. This study investigated the effect of thermal cycling on the microleakage at the interface of three hard chairside reline resins and three denture base resins. Material and methods: Rectangular bars (12 mm × 3 mm × 3 mm) of Lucitone 550, Acron MC and QC 20 were made and relined with Kooliner, Tokuyama Rebase Fast II and Ufi Gel Hard, Lucitone 550, Acron MC and QC 20 resins. Specimens were divided into one control and two test groups (n = 10). In specimens of the control group, the microleakage was performed after the reline procedure. In Test Group 1, the specimens were stored for 24 h in distilled water at room temperature and in Test Group 2; the specimens were thermal cycled from 5 to 55°C for 5000 cycles with a 30‐s dwell time. Subsequently, all specimens were immersed in 50% silver nitrate solutions for 24 h. All specimens were sectioned longitudinally into three fractions and the lateral sections were examined (n = 20). Silver nitrate stain penetration was examined under a stereoscopic lens with ×30 magnification, and the images were captured. Leica Qwin image analysis software was used to determine microleakage at the interface of the materials. Data were analysed using the Kruskal–Wallis test at a 95% level of significance. Results: For all cycles, there were no statistically significant differences between thermal cycled and non‐thermal cycled groups (p > 0.05). Conclusion: It can be concluded that thermal cycling had no effect on the microleakage.  相似文献   

6.
An archaeal ether-linked lipid, archaeol, was determined to be a biomass indicator for methanogens both in the laboratory enriched culture and in marine sediments. The archaeol measurement method described by Ohtsubo et al. in 1993 was modified and applied to marine sediments. We compared the amount of archaeol with the cell number of methanogens or methane concentration in laboratory enriched culture of methanogens from marine sediment. Good correlations were obtained as follows: (Methane, mmol) = 11.2 × (Archaeol, mg): r= .996 or (Cell number) = 1.13 × 1011× (Archaeol, mg): r= .995, respectively. In the sediments of Tokyo Bay, archaeol was measured from approximately 46 to 561 ng/dry g sediment at the entrance to 267 to 4160 ng/dry g sediment at the innermost area. Using the coefficient from the laboratory experiment, these data corresponded to cell numbers of 5.2 × 106 to 4.7 × 108/dry g sediment. These values were 1 or 2 orders of magnitude higher than those obtained by culture methods in previous studies. Although dead or decomposed cells might be detected, archaeol measurement is useful for estimating the biomass of methanogens because of the good correlation between methane concentration and archaeol content in marine environments. In this study, we found a correlation of (Methane, mmol) = 0.012 × (Archaeol, mg): r= .932, n= 17 in marine sediments. Received December 21, 1998; accepted June 16, 1999  相似文献   

7.
A sample composed of 4973 specimens of the spotted goatfish, Pseudupeneus maculatus (4.5–29.2 cm fork length) was collected off northeastern Brazil from 1999 to 2002. Sagittae (398) were analyzed for age and growth estimation using both annuli and microincrements. For the first approach, the mean monthly marginal increment ratio analysis suggested that one band is formed annually from May to June. With the microincrements analysis, 15 specimens were reared (3–71 days) for validation of periodicity using oxytetracycline. Fluorescent dyes were observed in 10 specimens (7.5–16.9 cm). After testing, one microincrement was found to form daily. Otolith sections were polished and analyzed using a light microscope (100–1200× magnifications). Individuals larger than 22.6 cm (4 years) showed heavily overlaid increments on the otolith edge, which hampered the counts. There was agreement on the size from counts on an annual and daily basis. The von Bertalanffy function provided the following parameters: L = 33.23 cm; K = 0.265; t0 = 0.0883 year. The sample was composed of 0+ to >5‐year‐old specimens. Age at first maturity was 3.6 years (20 cm), whereas the recruitment age for fisheries was 3.9 years. Juveniles represented about half of the catch (48.7%), which may have implications for population equilibrium.  相似文献   

8.
A flow injection method with chemiluminescence detection is reported for the determination of vitamin A. The method is based on the enhancement effect of vitamin A on chemiluminescence of tris(2,2′‐bipyridyl)Ru(II)–Ce(IV) in acidic medium. The proposed procedure is used to quantitate vitamin A in the range 1.0–100 × 10?6 mol/L with a correlation coefficient of 0.9991 (n = 9) and relative standard deviation in the range 1.2–2.3% (n = 4). The limit of detection (3 × blank) was 8.0 × 10?8 mol/L with a sample throughput of 100/h. The effect of common excipients used in pharmaceutical formulations and some clinically important compounds was also studied. The method was applied to determine vitamin A in pharmaceutical formulations and the results obtained were in reasonable agreement with the amount quoted. The results were compared using spectrophotometric method and no significant difference was found between the results of the two methods at 95% confidence limit. Copyright © 2009 John Wiley & Sons, Ltd.  相似文献   

9.
One of the most common and ubiquitous methods to age mammals is by counting the cementum annuli in molars, premolars, incisors, or canines. Despite the ubiquity and perceived simplicity of the method, cementum annuli analysis can be time-consuming, expensive, inaccurate, and imprecise, and require specialized equipment. Using beavers (Castor canadensis) as a test species, we developed a straightforward method to age mammals that requires little specialized equipment. The method consists of: (1) digitizing longitudinally sectioned teeth and measuring the proportion of tooth surface area comprised of cementum (“proportion cementum”), (2) evaluating the relationship between proportion cementum and specimen age (determined from either known-age samples or cementum annuli analysis), and (3) using the modeled relationship to estimate the age of other individuals based solely on proportion cementum. The relationship between proportion cementum and age was strongly correlated (R2 = .97–.98 depending on observer), similar between observers, and similar between known-age specimens and those aged via cementum annuli analysis. Using this proportion cementum method, two independent observers accurately predicted the age of 80%–84% of specimens within 0.5 year and 96%–98% within 1 year. We suggest this aging method will likely work with most mammal species given the relatively consistent deposition of cementum throughout mammals' lives and has promise to be a simple and quick alternative to cementum annuli analysis regardless of whether one develops proportion cementum models using known-age specimens or those aged via alternative methods.  相似文献   

10.
The method of polymerase chain reaction with random primers (RAPD PCR) was used to identify the progeny of the crosses between three sturgeon species, Amur sturgeon (Acipenser schrenckii Brandt, 1869), Siberian sturgeon (A. baerii Brandt, 1869), and sterlet (A. ruthenus Linnaeus, 1758). Using ten primers, genetic variation in 70 yearlings, produced in seven individual crosses: Acipenser schrenckii × A. schrenckii, A. baerii × A. baerii, A. ruthenus × A. ruthenus, A. schrenckii × A. baerii, A. baerii × A. schrenckii, A. schrenckii × A. ruthenus, and A. ruthenus × A. schrenckii was described and evaluated. It was demonstrated that the samples composed of hybrids from individual crosses were more variable than the samples of parental species. On the other hand, pooled samples of hybrids from two cross directions were genetically less variable than the pooled samples of their parents. The three main features of the hybrid RAPD profiles identified included: (1) preservation of marker DNA fragments of both parents in one genome; (2) presence of specific DNA fragments, absent from both parents; and (3) dependence of the frequency of some DNA fragments from the cross direction. Multidimensional scaling clearly distinguishes in the space of three coordinates the individuals of original species and the hybrid progeny with differentiation in the groups of direct and backcross hybrids. Analysis of relationships (UPGMA and NJ) pointed to substantial differentiation between the species, as well as between the species and hybrid progeny. Close genetic relationships between direct and backcross hybrids were demonstrated. Multilocus RAPD markers in association with statistical methods are considered to be the useful tool for discrimination of interspecific hybrids of sturgeon. Possible reasons for the differences in the hybrid RAPD profiles are discussed. Original Russian Text ? K.V. Rozhkovan, G.N. Chelomina, E.I. Rachek, 2008, published in Genetika, 2008, Vol. 44, No. 11, pp. 1453–1460.  相似文献   

11.
Crown and cusp areas of mandibular molars were measured and analyzed on a sample of 249 specimens attributed to Australopithecus afarensis, A. africanus, A. (Paranthropus) robustus, A. (P.) boisei, and early Homo. In addition to intertaxon comparisons, we compared data that had been collected independently by two of the authors using methods that differ slightly in technique of measurement. Interobserver differences were evaluated by the t-test of paired comparisons, method error statistic, percent differences, and principal component analysis. Results suggest that between-technique error of measurement of overall crown area is small. Error estimates for individual cusp area measurements were of larger relative magnitude. However, these were not sufficient to detract from the conclusions derived from comparative analyses. Our results are in general agreement with previous assessments of early hominid dental size. Crown areas of A. africanus, however, exhibit a mosaic pattern, with M1 similar in size to that of A. afarensis and early Homo, and M2 and M3 similar in size to that of A. robustus. Intertaxon comparisons of relative cusp area were undertaken by univariate statistics and principal component analysis. These analyses revealed that while A. (P.) robustus and A. (P.) boisei both possess mandibular molars with cusp proportions significantly different from the ‘non-robust’ taxa, these differences are substantially greater in A. (P.) boisei. © 1994 Wiley-Liss, Inc.  相似文献   

12.
Electron micrographs of Physarum polycephalum microplasmodia (LU887 × LU897) reveal cytoplasmic inclusions that appear “striated” at low magnifications; at higher magnifications these exhibit a structure that we have interpreted as microtubule bundles. The light and dark regions in the inclusions are due to the affinity of some microtubules for osmic acid; these appear to have dense regions while other microtubules remain electron lucent. The diameters of the microtubules are about 32–33nm; the subunits forming the tubule walls measure about 8–9nm in diameter. The diameter measurements are slightly larger than the dimensions assigned to vertebrate microtubules (28nm); however, the diameter of the subunits in the microtubule wall measures about 8–9nm which is essentially the same measurement reported for vertebrate tubulin dimers.  相似文献   

13.
The Iberian mountain spiny fescues are a reticulate group of five diploid grass taxa consisting of three parental species and two putative hybrids: F. × souliei (F. eskia × F. quadriflora) and F. × picoeuropeana (F. eskia × F. gautieri). Phenotypic and molecular studies were conducted with the aim of determining the taxonomic boundaries and genetic relationships of the five taxa and disentangling the origins of the two hybrids. Statistical analyses of 31 selected phenotypic traits were conducted on individuals from 159 populations and on nine type specimens. Molecular analyses of random amplified polymorphic DNA (RAPD) markers were performed on 29 populations. The phenotypic analyses detected significant differences between the five taxa and demonstrated the overall intermediacy of the F. × picoeuropeana and F. × souliei between their respective parents. The RAPD analysis corroborated the genetic differentiation of F. eskia, F. gautieri and F. quadriflora and the intermediate nature of the two hybrids; however, they also detected genetic variation within F. × picoeuropeana. These results suggest distinct origins for F. × picoeuropeana in the Cantabrian and Pyrenean mountains, with the sporadic Pyrenean populations having potentially resulted from recent hybridizations and the stabilized Cantabrian ones from older events followed by potential displacements of the parents. © 2013 The Linnean Society of London, Botanical Journal of the Linnean Society, 2013, 173 , 676–706.  相似文献   

14.
An essential pilot study was designed to quantify observer heterogeneity and to compare observation methods for the detectability of forest birds in stands of Eucalyptus and Pinus radiata forest as a basis for a major research project on habitat fragmentation near Tumut, southern New South Wales. Twelve experienced observers participated in the investigation. Point interval counts, zig-zag walks and strip transects were used to count birds in both eucalypt and pine forests. The 65 species of birds recorded in the study were assigned to one of nine groups classified by a set of attributes that characterized bird detection by field observers (e.g. body size, colour and calling patterns). Observer heterogeneity varied between groups of birds and was most apparent for small birds foraging in low shrubs (species such as the white-browed scrub wren, assigned to group 2), frequent calling, active birds (species such as the golden whistler, assigned to group 7), and midstorey, undercanopy foragers with distinctive behaviour (species such as the grey fantail assigned to group 4). For bird groups 2, 4 and 7, additional variability due to observer differences resulted in an average increase of ~ 40% in the width of a 95% confidence interval for the logarithm of bird abundance generated from a 20 minute count. Our analysis shows that taking the average of counts obtained by two or more observers would negate the increase in variance of counts due to observer heterogeneity. Few differences between methods of field observation were found. However, for frequent calling, active birds (group 7) there was evidence that more birds were heard using the point interval count method. Our study clearly demonstrated a need to either control for observer differences or to assign at least two observers to individual sites when designing bird surveys for comparative studies. Failure to do so will result in a decrease in precision of bird counts.  相似文献   

15.
The projected structures of two unstained periodic biological specimens, the purple membrane and catalase, have been determined by electron microscopy to resolutions of 7 Å and 9 Å, respectively. Glucose was used to facilitate their in vacuo preservation and extremely low electron doses were applied to avoid their destruction.The information on which the projections are based was extracted from defocussed bright-field micrographs and electron diffraction patterns. Fourier analysis of the micrograph data provided the phases of the Fourier components of the structures; measurement of the electron diffraction patterns provided the amplitudes.Large regions of the micrographs (3000 to 10,000 unit cells) were required for each analysis because of the inherently low image contrast (<1%) and the statistical noise due to the low electron dose.Our methods appear to be limited in resolution only by the performance of the microscope at the unusually low magnifications which were necessary. Resolutions close to 3 Å should ultimately be possible.  相似文献   

16.
Abstract

The geomagnetic field effect hypothesis and the variability among replications in germination and growth tests in Triticum. – During previous eperiments on germination and growth of Triticum, we have often found many cases of heteroscedasticity and of statistically significant differences among contemporary replications (i.e. germinators). In such experiments all the caryopses were oriented in the same way inside the germinators, whilst the germinators were randomly set inside the thermostats: therefore the seeds of different germinators might result differently oriented. Some researchers report that seeds differently oriented according to the lines of force of the geomagnetic field, may respond with different germination, growth rates and growth directions of shoots and roots (magnetotropism). The present investigation was designed to ascertain if these effects might be responsible of the variability among the contemporary replications. The different caryopses orientation within laboratory geomagnetic field, does not give evidence of any effect on germination, growth rate and direction. The caryopses show no germination sensibility neither related to different age, light and temperature conditions, neither to different cultivars, neither to varying orientation from the geomagnetic North every 15° and to varying time of ? preorientation ? in dry conditions. Differences between the variability of randomly and North oriented subsamples are not observed. No statistically significant differences are observed among shoots and roots lenghts of seedlings from differently oriented caryopses neither in different environmental conditions, nor as function of the orientation (24 angles of 15°), nor after different preorientations, nor after repeating the test on 608 subsamples (12.000 seedlings). Therefore the variability among contemporary replications is not imputable to the geomagnetic field. The variability and the distributions structure analysis suggest that both the heteroscedasticity and the significant differences among the replications may be due to an interaction among the caryopses inside the germinators through the substrate. This is probably related to the cultivar characteristics. We emphasize that an accurate evaluation of the variability is particularly necessary in germination and growth tests.  相似文献   

17.
Summary

A study was made in the Cairngorms, Scotland to make recommendations for a monitoring scheme capable of detecting changes in the vegetation caused by recreational pressure following the development of a funicular railway. Four methods were used in field trials to assess percentage cover of plant species and gravel, rock and bare ground, where appropriate, in two vegetation types (open and closed). The methods used were visual estimates in 50 × 40 cm quadrats (Q), the mean of visual estimates in twenty 10 × 10 cm sub-quadrats of the 50 × 40 cm quadrats (Q20), a modified point intercept method (RL) and photography. Variances between observers and between-quadrats were estimated for the different methods. The sampling design for detecting change was based on a model of variance, constructed from field trial data.

Between-observer and between-quadrat variances were related to mean percentage cover and approximated to a binomial distribution. The between-quadrat variance was larger than observer variance. The Q20 method achieved appreciably better precision than the other methods. Analysis of half of the 10 × 10 cmsub-quadrats (1/2Q20) selected in a checker board design achieved a relative efficiency of 78% compared with the Q20. This result suggests that comparable precision to the Q20 method could be achieved by choosing about 14 sub-quadrats in a larger quadrat, thus saving some time. Variation between quadrats also suggested that the Q20 method was the one of choice for maximising precision. The precision of the photographic method was based on fewer data points, so is less accurate than other estimates.

Minimum sample sizes were estimated for detecting a 10% relative change of a species in open vegetation with 30% cover (i.e. a change from 30% to <27 or to >33% cover). With a 10 % Type II error rate and 5 % Type I error rate the minimum sample sizes were 47 quadrats for Q, 18 for Q 20, 43 for RL, and 23 for the means of ten 10 × 10 cm sub-quadrats in open vegetation.

The most time-efficient field recording appeared to be the use of Q despite the required sample size being 2.6 times higher than that of Q20. The far lower time requirement per quadrat, however, compensated for the higher numbers. The number of quadrats would depend on the specified change in percentage cover and on the statistical significance level used. For example, to detect a 10% absolute change in cover (i.e. from 30% to either <20 % or >40 % cover) at 95 % probability the net effective recording time is estimated at 5 h per vegetation type while to detect a 5 % change at 99 % probability would require c. 25 h. Larger samples may be required for other species or for species with a low initial cover.  相似文献   

18.
The Xenarthra represents an enigmatic clade of placental mammals that includes living tree sloths, armadillos, and their extinct relatives, yet certain aspects of the biology of this group remains poorly understood. Here, we use scanning electron microscopy to test the hypothesis that orthodentine microwear patterns in extant xenarthrans are significantly different among different dietary groups. In a blind analysis, microwear patterns were quantified at a magnification of 500× by two independent observers for extant species from four dietary groups (carnivore–omnivores, folivores, frugivore–folivores, and insectivores). Independent observers recovered the same relative between‐group differences in microwear patterns. Insectivores and folivores have a significantly lower numbers of scratches and greater scar widths than frugivore–folivores and carnivore–omnivores, yet we were neither able to statistically distinguish insectivores from folivores, nor differentiate frugivore–folivores from carnivore–omnivores. Nevertheless, a clear distinction exists between taxa from the same trophic level and habitat, which suggests that orthodentine microwear reflects niche partitioning and habitat more than diet among related forms. We suggest that bite force and chewing mechanics have a strong influence on the formation of orthodentine microwear, which may explain some of the observed overlap between distinct groups (e.g. frugivore–folivores versus carnivore–omnivores). This study serves as a positive step forwards in our understanding of the ecological role of living xenarthrans, and serves as a foundation for using orthodentine microwear to reconstruct palaeoecology in extinct ground sloths, glyptodonts, and pampatheres. © 2012 The Linnean Society of London, Biological Journal of the Linnean Society, 2012, ?? , ??–??.  相似文献   

19.
doi: 10.1111/j.1741‐2358.2010.00379.x
Effectiveness of different cleaning agents on the adherence of Candida albicans to acrylic denture base resin Objective:  To evaluate the ability of three alkaline peroxide‐type (Polident, Efferdent, Fittydent) and two mouth rinse cleaning agents (CloSYSII and Corsodyl) to inhibit Candida albicans on acrylic denture base resin. Background:  Appropriate routine cleaning of dentures is necessary to prevent denture stomatitis and maintenance of healthy supporting tissues. Materials and methods:  A total of 180 acrylic resin specimens (10 × 10 × 2 mm) were prepared and divided into six groups. Candida albicans was incubated on Sabouraud dextrose agar (SDA) at 37°C for 48 h. After dilution, a final yeast suspension of approximately 10 6 C. albicans per millimetre was prepared. Ten acrylic resin specimens for each group were placed in a sterile Petri dish covered with 20 ml of fungal suspension and incubated at 37°C for 90 min. Then, the specimens were immersed in 40 ml of the test solution at 37°C for 15, 30 and 60 min. Fungal cells adhering to acrylic resin surfaces were fixed in formaldehyde and counted microscopically. Results:  Mouth rinses showed the highest removal activity for all the treatment times and completely eliminated the adherence of C. albicans. Conclusions:  The use of mouth rinse may be a suitable method for cleaning dentures.  相似文献   

20.
《Animal behaviour》1986,34(4):1016-1025
Fourteen adult female domestic cats were watched by two observers for 3 months. Ratings of 18 aspects of each cat's behavioural style were obtained independently from each observer. Correlations between observers were statistically significant for 15 of the 18 aspects and seven of the correlation coefficients were greater than 0·7. The ratings were compared with results of direct recording methods, where equivalent measures were available and, in five out of six cases, the results of the ratings and direct methods were significantly correlated. The rating method is, therefore, generally reliable and can be adequately validated. Some assessments of observer ratings which are not obviously and easily related to direct recordings may prove particularly useful in developmental studies of alternative modes of behaviour and the origins of individual differences.  相似文献   

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