bluuebunny/crossref_metadata_2025 huggingface dataset:
Techniques for rapid quantitative assessment of activity levels in small‐group tutorials
Authors: Prinz  , Yip  , Tipoe  , Lucas  , Lenstrup
Published: March 2003

Two techniques for the rapid quantitative analysis of student participation in small‐group teaching were investigated. In the first approach an observer, who also acted as a `critical friend', recorded the length of individual contributions using a computer keyboard as a simple timing device. In the second approach, small‐group sessions were recorded with a portable stereophonic audiotape recorder. The teacher was recorded on one channel, all students on the other. A computer program produced automated analysis of these small group interactions by computing relative amount of speech on each channel. Simple analysis produced automatically by the programs revealed the overall style of the tutorial – variably `mini‐lectures' by teachers with very little participation by the student body, rapid `question and answer' sessions with about equal teacher/student body involvement or `mini‐presentations' by students with the teacher offering sparse comments in the manner of a facilitator. By presenting results in a graphic format, teachers can be given rapid objective feedback on their teaching style. Coupled with short verbal/non‐verbal quizzes at the end of tutorials and information from other assessments, the value of using levels of participation as a measure of the efficiency of such small‐group sessions can itself be assessed.
Course in massage therapy for medical students
Authors: Falkenbach  , Blumenthal  , Bühring
Published: March 2003

Massage courses for medical students have been held at Frankfurt University Medical School since 1987. To evaluate the motives for participation and to record possible changes in the attitude towards massage therapy, the students were asked to fill out a standardized questionnaire in 1990, 1993 and 1995/96. The results show that the motive for participation and the attitude towards massage therapy remained widely unchanged during these years. Summarizing all data (n = 199) the motives for participation were: (1) to practise massage therapy (86%), (2) to be better able to (later) prescribe massage therapy (66%), (3) to improve palpation skills (75%), (4) to do `something practical' (56%), and (5) to (later) practise massage therapy as a medical doctor (23%). On average, the proportion of theory and practical instruction of 1:3·2 was considered suitable.
Psychiatric training for family doctors: what do GP registrars want and can a brief course provide this?
Authors: Ratcliffe  , Gask  , Creed  , Lewis
Published: March 2003

Context

About 40% of British General Practitioners (GPs) train formally in a psychiatric post as part of their general practice training, but such training may not fully meet the needs of future GPs. A specific course in psychiatry for family doctors has run in Manchester for more than a decade.

Method

Semi‐structured interviews conducted with GP registrars before attending the Manchester course in psychiatry with questionnaire follow‐up afterwards to ascertain (a) the training ‘wants’ of GP registrars and (b) whether the course was providing them.

Results

GP registrars most frequently wanted training in communication skills, how to access the resources that are available to GPs, the detection of psychiatric illness, drug treatment and the management of aggression. The course was successful in satisfying the first three but failed in the last two. There was trend for those who attended Manchester Medical School, which scored significantly higher on number of topics covered at undergraduate level, to perceive a greater need for training than those who attended other medical schools. However, there was no evidence to link self‐perception of greater need with having already worked in general practice during postgraduate training.

Conclusions

More attention needs to be paid to how to address the specific mental health skills training requirements of GP registrars both within the attachment in psychiatry and during the practice year. Preliminary research is required to devise teaching packages before they are entirely satisfactory for GP education.
Portfolios in continuing medical education – effective and efficient?
Authors: Mathers N J , Challis M C , Howe A C , Field N J
Published: March 2003

Introduction

A cross over comparison between ‘traditional’ continuing medical education (CME) activities and portfolio‐based learning in general practice is described.

Method

Thirty‐two volunteer general practitioners (GPs) were divided into two cohorts; each cohort spent six months following a ‘traditional’ route to postgraduate educational accreditation (PGEA) and six months following a portfolio‐based learning route supported by three CME tutors.

Outcome Measures

These were the submission of a completed portfolio with evidence of the completion of learning cycles and participants reflections on the educational process. Qualitative and quantitative evaluation data were collected by questionnaire, semi‐structured interview, participant observation and review of completed portfolios.

Results

The themes identified by GPs as hopes for the portfolios were largely fulfilled and the anxieties generally confounded. The flexibility of the portfolio learning process was particularly important to the participants. The breadth of topics covered by the portfolios was extremely wide and comparison with the submissions for ‘traditional’ PGEA showed a much smaller spread of learning activities and fewer subjects of study.

Effectiveness

The use of the portfolios of critical incidents and the completion of learning cycles with application to practice provided evidence of the effectiveness of such learning.

Efficiency

The mean number of hours spent by GPs preparing the portfolios was 24.5 ± 12 (SD) which was significantly more than the 15 hours of PGEA awarded.

Conclusions

This study demonstrates that a portfolio‐based learning scheme can meet the needs of GPs relevant to their professional practice; it can give learners control over how, what and when they learn and encourage active and peer‐supported learning; it can build personal and professional confidence and be thought both valid and reliable by participants. Learning outcomes can also be reliably assessed by PGEA within the context of an individually created learning plan.
Educational portfolios in the assessment of general practice trainers: reliability of assessors
Authors: Pitts John , Coles Colin , Thomas Peter
Published: March 2003

Objectives

This paper reports a project that assessed a series of portfolios assembled by a cohort of participants attending a course for prospective general practice trainers.

Design

The reliability of judgements about individual ‘components’, together with an overall global judgement about performance were studied.

Setting

NHSE South & West, King Alfred’s College, Winchester and Institute of Community Studies, Bournemouth University.

Subjects

Eight experienced general practice trainers recruited from around Wessex, which incorporates Hampshire, Dorset, Wiltshire and the Isle of Wight.

Results

The reliability of individual assessor’s judgements (i.e. their consistency) was moderate, but inter‐rater reliability did not reach a level which could support making a safe summative judgement. The levels of reliability reached were similar to other subjective assessments and perhaps reflected individuality of personal agendas of both the assessed and the assessors, and variations in portfolio structure and content.

Conclusions

Suggestions for approaches in future are made.
Gene flow among Tetranychus urticae (Acari: Tetranychidae) populations in Greece
Authors: TSAGKARAKOU ANASTASIA , NAVAJAS MARIA , PAPAIOANNOU‐SOULIOTIS PAGONA , NICOLE PASTEUR†
Published: March 2003

Polymorphism of four enzymatic loci has been examined in 27 populations of Tetranychus urticae in relation to their geographical distribution and to two ecological parameters: open field vs. greenhouse habitats, and species of the colonized host plant. Genetic differentiation was significantly correlated to geographical distance in both types of habitat. Mite density and distribution of infested plants appear to be important factors for the population structure of T. urticae. In open field, T. urticae specimens from citrus trees were genetically more similar to other ‘citrus’ samples collected in different localities than they were from mites collected in the same locality on other plant species.
Phylogeographic studies in plants: problems and prospects
Authors: SCHAAL B. A. , HAYWORTH D. A. , OLSEN K. M. , RAUSCHER J. T. , SMITH W. A.
Published: March 2003

Genetic structuring of plant populations is strongly influenced by both common ancestry and current patterns of interpopulation genetic exchange. The interaction of these two forces is particularly confounding and hence interesting in plants. This complexity of plant genetic structures is due in part to a diversity of reproductive ecologies affecting genetic exchange and the fact that reproductive barriers are often weak between otherwise morphologically well‐defined species. Phylogeographic methods provide a means of examining the history of genetic exchange among populations, with the potential to distinguish biogeographic patterns of genetic variation caused by gene flow from those caused by common ancestry. With regard to plants, phylogeography will be most useful when applied broadly across the entire spectrum of potential genetic exchange. Although current phylogeographic studies of plants show promise, widespread application of this approach has been hindered by a lack of appropriate molecular variation; this problem is discussed and possible solutions considered.
Comparative mtDNA phylogeography of neotropical freshwater fishes: testing shared history to infer the evolutionary landscape of lower Central America
Authors: BERMINGHAM E. , MARTIN A. P.
Published: March 2003

Historical biogeography seeks to explain contemporary distributions of taxa in the context of intrinsic biological and extrinsic geological and climatic factors. To decipher the relative importance of biological characteristics vs. environmental conditions, it is necessary to ask whether groups of taxa with similar distributions share the same history of diversification. Because all of the taxa will have shared the same climatic and geological history, evidence of shared history across multiple species provides an estimate of the role of extrinsic factors in shaping contemporary biogeographic patterns. Similarly, differences in the records of evolutionary history across species will probably be signatures of biological differences. In this study, we focus on inferring the evolutionary history for geographical populations and closely related species representing three genera of primary freshwater fishes that are widely distributed in lower Central America (LCA) and northwestern Colombia. Analysis of mitochondrial gene trees provides the opportunity for robust tests of shared history across taxa. Moreover, because mtDNA permits inference of the temporal scale of diversification we can test hypotheses regarding the chronological development of the Isthmian corridor linking North and South America. We have focused attention on two issues. First, we show that many of the distinct populations of LCA fishes diverged in a relatively brief period of time thus limiting the phylogenetic signal available for tests of shared history. Second, our results provide reduced evidence of shared history when all drainages are included in the analysis because of inferred dispersion events that obscure the evolutionary history among drainage basins. When we restrict the analysis to areas that harbour endemic mitochondrial lineages, there is evidence of shared history across taxa. We hypothesize that there were two to three distinct waves of invasion into LCA from putative source populations in northwestern Colombia. The first probably happened in the late Miocene, prior to the final emergence of the Isthmus in the mid‐Pliocene; the second was probably coincident with the rise of the Isthmus in the mid‐Pliocene, and the third event occurred more recently, perhaps in the Pleistocene. In each case the geographical scale of the dispersion of lineages was progressively more limited, a pattern we attribute to the continuing development of the landscape due to orogeny and the consequent increase in the insularization of drainage basins. Thus, the fisheye view of LCA suggests a complex biogeographic history of overlaid cycles of colonization, diversification, sorting and extinction of lineages.
Host‐based genetic differentiation in the aphid Aphis gossypii Glover, evidenced from RAPD fingerprints
Authors: VANLERBERGHE‐MASUTTI FLAVIE , CHAVIGNY PASCAL
Published: March 2003

Samples of the aphid Aphis gossypii (Glover) were collected from different host plants at 18 locations in southern France, La Réunion, Portugal and Laos. RAPD (random amplified polymorphic DNA) patterns of the 480 aphids were obtained using three random primers. A large number of RAPD bands were shared by all aphids of the 18 populations, but some RAPD bands appeared to be population specific. Over all aphids, a total of 37 polymorphic bands were identified and defined 142 RAPD phenotypes. A cluster analysis based on genetic distance revealed that the 18 aphid populations were divided into two groups, depending on whether they were collected on a cucurbit host plant. An analysis of molecular variance (AMOVA) was also performed and confirmed the differentiation into two groups. Several RAPD bands that were obtained using random primer A11 could be considered as diagnostic loci as they were fixed in populations collected on cucurbits and were always absent in those collected on noncucurbit host plants. These results represent the first evidence for genetic structuring within the species A. gossypii, according to host‐plant type.
Gene flow between insular, coastal and interior populations of brown bears in Alaska
Authors: PAETKAU DAVID , SHIELDS GERALD F. , STROBECK CURTIS
Published: March 2003

The brown bears of coastal Alaska have been recently regarded as comprising from one to three distinct genetic groups. We sampled brown bears from each of the regions for which hypotheses of genetic uniqueness have been made, including the bears of the Kodiak Archipelago and the bears of Admiralty, Baranof and Chichagof (ABC) Islands in southeast Alaska. These samples were analysed with a suite of nuclear microsatellite markers. The ‘big brown bears’ of coastal Alaska were found to be part of the continuous continental distribution of brown bears, and not genetically isolated from the physically smaller ‘grizzly bears’ of the interior. By contrast, Kodiak brown bears appear to have experienced little or no genetic exchange with continental populations in recent generations. The bears of the ABC Islands, which have previously been shown to undergo little or no female‐mediated gene flow with mainland populations, were found not to be genetically isolated from mainland bears. The data from the four insular populations indicate that female and male dispersal can be reduced or eliminated by water barriers of 2–4 km and 7km in width, respectively.
Polymorphic microsatellite markers for population analysis of a tephritid pest species, Bactrocera tryoni
Authors: KINNEAR M. W. , BARIANA H. S. , SVED J. A. , FROMMER M.
Published: March 2003

To obtain a set of microsatellite markers for the Queensland fruit fly Bactrocera tryoni, a genomic library was screened with a number of simple repeat oligonucleotide probes. Sequencing recovered 22 repeat loci. The microsatellite sequences were short, with repeat numbers ranging from five to 11. Of these, 16 polymerase chain reaction (PCR) primer sets yielded amplifiable products, which were tested on 53 flies from five widely separated sites. All loci showed polymorphism in the population sample, with the number of alleles ranging from two to 16. Several dinucleotide repeats showed alleles separated by single‐base differences and multiple steps, suggesting a mutation process more complex than the stepwise mutation model.
Queen mating and paternity variation in the antLasius niger
Authors: BOOMSMA J. J. , VAN DER HAVE T. M.
Published: March 2003

We have electrophoretically analysed the variation in queen mating and worker paternity across and within seven populations of the antLasius nigerin northwestern Europe. Populations were panmictic and not genetically differentiated (FST= 0.003 ± 0.004; rangec.1000 km). Queens (n= 535) were shown to mostly mate with a single male, but double mating occurred in all populations and triple mating was found in one case (the total number of worker offspring analysed for paternity was 4825). The genetically effective queen mating frequency was 1.16 on average across populations (range 1.04–1.42). Double sampling of six out of the seven populations showed that most of the variation in queen mating occurred among populations and not within populations among years. Also, paternity skew in colonies with double‐mated queens was relatively constant per site but varied across populations. Paternity skew was high in populations with low frequencies of double queen‐mating, low in populations with intermediate frequencies of double queen mating and ambiguous in a single population where more than half of the queens mated multiply. Double‐mated queens were only collected halfway through a nuptial flight, suggesting that double mating is time consuming and that the mating swarm sex ratio may affect the likelihood of multiple mating towards the end of a flight. No difference in fresh weight between single‐ and double‐mated queens from the same population was found.
Stability of population structure and genetic diversity across generations assessed by microsatellites among sympatric populations of landlocked Atlantic salmon (Salmo salar L.)
Authors: TESSIER NATHALIE , BERNATCHEZ LOUIS
Published: March 2003

It may often be necessary to perform genetic analyses of temporal replicates to estimate the significance of spatial variation independently from that of temporal variation in order to ensure the reliability of estimates of a defined population structure. Nevertheless, temporal studies of genetic diversity remain scarce in the literature relative to the plethora of empirical studies of population structure. In vertebrates, a limited number of studies have specifically assessed the temporal stability of population structure for more than one generation. In this study, we performed a microsatellite analysis of DNA obtained from archived scales to compare the population structure among four sympatric landlocked populations of Atlantic salmon (Salmo salar) over a time frame of three to five generations. The same patterns of allele frequency distribution, θ, RST and genetic distance estimates were observed among populations for two time periods, confirming the temporal stability of the population structure. Despite population declines and stocking during this period, no statistically significant changes in intrapopulation genetic diversity were apparent. This study illustrates the feasibility and usefulness of microsatellite analysis of temporal samples, not only to infer changes of intrapopulation genetic diversity, but also to assess the stability of population structure over a time frame of several generations.
Do assemblages of Coregonus (Teleostei: Salmoniformes) in the Central Alpine region of Europe represent species flocks?
Authors: Douglas M. R. , Brunner P. C. , Bernatchez L.
Published: March 2003

To examine models of evolution for Coregonus from the Central Alpine region of Europe, 20 populations from nine lakes were assessed for variation at six microsatellite DNA loci. Patterns of variation were tested against three evolutionary models: phenotypic plasticity, multiple invasions of lakes by divergent forms, and within‐lake radiation of species flocks. All sympatric and all but one allopatric pairs of populations were significantly divergent in allele frequencies. Pairwise F‐statistics indicated reduced gene flow among phenotypically divergent sympatric populations. These results reject the hypothesis that within‐lake morphological and ecological diversity reflects phenotypic plasticity within a single gene pool. Genetic similarity was higher among forms within lakes than between populations of the same form in different lakes. Among‐lake divergence was primarily a product of allele size differences. Mantel tests contrasting patterns of genetic divergence against patterns predicted from the multiple invasions and species flocks models indicated that the latter is the best explanation of the observed genetic variation. Thus, reproductively isolated species diverged within lakes, with similar patterns repeatedly emerging among lakes. While this study argues for a particular mode of evolution in Central Alpine Coregonus, the taxonomy of these forms remains unresolved.
Promiscuity in populations of the cushion plant Saxifraga oppositifolia in the Swiss Alps as inferred from random amplified polymorphic DNA (RAPD)
Authors: GUGERLI F. , EICHENBERGER K. , SCHNELLER J. J.
Published: March 2003

Overviews on patterns of genetic variation within and among plant populations show that widespread, outcrossing species should have a high proportion of the total genetic variation within populations and a low proportion among populations, which results in little population differentiation. However, in Alpine areas, large–scale distribution barriers as well as small‐scale habitat heterogeneity could lead to geographical and temporal isolation, respectively. We investigated the genetic variation of Saxifraga oppositifolia from 10 populations of the Alps in southeastern Switzerland using random amplified polymorphic DNA (RAPD). Based on the banding patterns of four RAPD primers, 84 polymorphic markers identified all 189 sampled individuals as being genetically different. The genetic variation was mainly found within populations (95%), whereas less than 5% was found among populations and among regions. Analyses of molecular variance (AMOVA) suggested that population differentiation was highly significant. However, grouping populations differently into regions did not appear to result in a clear correspondence of genetic and geographical relatedness. Genetic variation did not significantly differ between populations of two elevational levels. This coincides with results of former pollination experiments that revealed a breeding system of S. oppositifolia which remains the same irrespective of the elevation. We assume that the high outcrossing rate, rare clonal reproduction, and some long‐distance dispersal even among topographically separated populations are the crucial determinants for the pattern of genetic variation found in the investigated area.
Mitochondrial DNA variation in bull trout (Salvelinus confluentus) from northwestern North America: implications for zoogeography and conservation
Authors: Taylor Eric B. , Pollard Susan , Louie Derek
Published: March 2003

Bull trout, Salvelinus confluentus (Salmonidae), are distributed in northwestern North America from Nevada to Yukon Territory, largely in interior drainages. The species is of conservation concern owing to declines in abundance, particularly in southern portions of its range. To investigate phylogenetic structure within bull trout that might form the basis for the delineation of major conservation units, we conducted a mitochondrial DNA (mtDNA) survey in bull trout from throughout its range. Restriction fragment length polymorphism (RFLP) analysis of four segments of the mtDNA genome with 11 restriction enzymes resolved 21 composite haplotypes that differed by an average of 0.5% in sequence. One group of haplotypes predominated in ‘coastal’ areas (west of the coastal mountain ranges) while another predominated in ‘interior’ regions (east of the coastal mountains). The two putative lineages differed by 0.8% in sequence and were also resolved by sequencing a portion of the ND1 gene in a representative of each RFLP haplotype. Significant variation existed within individual sample sites (12% of total variation) and among sites within major geographical regions (33%), but most variation (55%) was associated with differences between coastal and interior regions. We concluded that: (i) bull trout are subdivided into coastal and interior lineages; (ii) this subdivision reflects recent historical isolation in two refugia south of the Cordilleran ice sheet during the Pleistocene: the Chehalis and Columbia refugia; and (iii) most of the molecular variation resides at the interpopulation and inter‐region levels. Conservation efforts, therefore, should focus on maintaining as many populations as possible across as many geographical regions as possible within both coastal and interior lineages.
Molecular anatomy of an onychophoran: compartmentalized sperm storage and heterogeneous paternity
Authors: Curach N. , Sunnucks P.
Published: March 2003

Onychophorans (peripatus or velvet worms) show extraordinarily high local endemism, and cryptic species are common. As part of a programme addressing issues of endemicity at hierarchical spatial scales, we investigated reproduction in Euperipatoides rowelli (Onychophora: Peripatopsidae) using microsatellite analysis. This species is ovoviviparous, and females have up to 70 embryos in their uteri simultaneously. Batches of undeveloped and well‐developed embryos may be present in the uteri of a female. Paired ovaries lead via a common oviduct into paired uteri, each of which has a spermatheca (sperm storage organ). Insemination in E. rowelli is dermal‐haemocoelic: spermatophores are placed on the skin of the female, the body wall is breeched, and sperm are released into the haemocoel through which they migrate to the spermathecae. There is no obvious mechanism to prevent sperm mixing, yet microsatellite analysis indicated that offspring in a female’s paired reproductive tracts can be sired by different males, and that the paired spermathecae can contain sperm from different males. More than 70% of females had broods with multiple paternity. The data are consistent with the potential for female postcopulatory influence over fertilizations: in particular, compartmentalization of sperm from different males into different spermathecae. Female control of fertilizations could lead to benefits including increased diversity of offspring, minimization of maternal–paternal genetic incompatibility, and influence on offspring genotypes. Multiple mating alone may increase the genetic diversity of offspring: this could be of importance in E. rowelli, which has very small genetic neighbourhoods and low genetic marker diversity.
Amplified fragment length polymorphism (AFLP) markers reveal extra‐pair parentage in a bird species: the bluethroat (Luscinia svecica)
Authors: Questiau S , Eybert M.‐C , Taberlet P
Published: March 2003

We tested the use of amplified fragment length polymorphism (AFLP) to assess the frequency of extra‐pair parentage in a bluethroat (Luscinia svecica namnetum) population. Thirty‐six families totalling 162 nestlings were analysed. Using a combination of three primer pairs, we reached an exclusion probability of 93% for the population. This probability can reach 99% considering families independently. We revealed that extra‐pair fertilizations are very common: 63.8% of all broods contain at least one extra‐pair young, totalling 41.9% of all young analysed. However, with the technique and the three primer pairs used it was not possible to attribute the parentage exclusions to extra‐pair paternity, maternity or both. As brood parasitism has never been reported in this species, it seems likely that the exclusions are due to extra‐pair males. This study shows that dominant AFLP markers can be useful for studying the mating system of taxa for which no microsatellite primers are available. This technique allows the approximate estimation of parentage exclusions despite the fact that it is not possible to know which parent has to be excluded.
Development of sequence amplified characterized region (SCAR) markers of Helicoverpa armigera: a new polymerase chain reaction‐based technique for predator gut analysis
Authors: Agustí N. , De Vicente M. C. , Gabarra R.
Published: March 2003

A method is described for the development of DNA markers for detection of Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) in predator gut analysis, based on sequence characterized amplified regions (SCARs) derived from a randomly amplified polymorphic DNA (RAPD) band. A 1200‐bp DNA fragment of H. armigera, absent in the predator band pattern and in other closely related prey species, was identified by RAPD analysis. This fragment was cloned and its extremes sequenced to design extended strand‐specific 20‐mer oligonucleotide primers. Three pairs of SCAR primers, which amplified three different DNA fragments, were used to study the effect of fragment length on detection of prey in the predator gut. Using the pair of primers that amplified the longest fragment of H. armigera DNA, a single band of 1100 bp was obtained, but its detection was not possible in the predator gut. Detection of the ingested prey was possible with the other two pairs of SCAR primers, obtaining bands of 600 and 254 bp, respectively. Detection of H. armigera DNA in the gut of the predator Dicyphus tamaninii was evaluated immediately after ingestion (t = 0) and after 4 h. Detection of H. armigera DNA after 4 h was only possible using the pair of primers that amplified the shortest fragment (254 bp). The test for specificity, using these last pair of primers, showed that H. armigera was the only species detected. The detection threshold was defined at a 1:8192 dilution of a H. armigera whole egg in all samples.
Identification of random amplified polymorphic DNA (RAPD) markers highly linked to sex determination in the red alga Gracilaria gracilis
Authors: Martinez E. A. , Destombe C. , Quillet M. C. , Valero M.
Published: March 2003

The random amplified polymorphic DNA (RAPD) technique was employed in the haplo‐diploid dioecious species Gracilaria gracilis to identify sex‐linked PCR markers. Sixty‐nine decamer oligonucleotide primers were tested on two bulks of DNA, one from five haploid males and the other from five haploid females. One of these primers (OPD13) generated a 430‐bp fragment specific to males and a 620‐bp fragment specific to females. The diploid individuals (tetrasporophytes) showed the co‐occurrence of these two fragments. In order to verify the linkage between the sexual phenotypes and these markers, a progeny array of 59 haploid individuals (male and female) born on a diploid individual was analysed, in all of which the two markers produced by the OPD13 primer segregated perfectly with sex.
Comparisons of mitochondrial DNA (mtDNA) sequences (16S rRNA gene) between oviparous and viviparous strains of Lacerta vivipara: a preliminary study
Authors: Heulin B. , Surget‐groba Y. , Guiller A. , Guillaume C. P. , Deunff J.
Published: March 2003

The lizard Lacerta vivipara has allopatric oviparous and viviparous populations. The mitochondrial DNA (mtDNA) gene coding for the 16S rRNA was sequenced for several viviparous lizard populations from France, Switzerland, Bulgaria, Czech Republic, The Netherlands, Sweden, and for oviparous lizard populations from the Pyrenean and Cantabric Mountains. Seven distinct groups (three oviparous and four viviparous) were identified. The net nucleotide divergence between oviparous and viviparous haplotypes was 1.3% ± 0.5 (mean ± standard deviation). These results on mtDNA, together with other data obtained previously, led us to formulate a biogeographical scenario that could be tested by further research.
Species distinction and evolutionary relationships of the Italian hare (Lepus corsicanus) as described by mitochondrial DNA sequencing
Authors: Pierpaoli M. , Riga F. , Trocchi V. , Randi E.
Published: March 2003

The taxonomic status of the Italian hare Lepus corsicanus has been uncertain since its first description by W. E. de Winton in 1898 (de Winton WE. Annual Magazine of Natural History, London, 1898, 1, 149–158). The distribution range of this species has shrunk severely over the last few decades owing to overhunting and restocking with nonindigenous brown hares (L. europaeus) in central and southern Italy and Sicily. Recently, scanty populations of Italian hares were rediscovered, and samples for morphological and molecular analyses were collected. Nucleotide sequences of the mitochondrial DNA (mtDNA) control region and cytochrome b indicate that L. corsicanus is a phylogenetically distinct species, which can be identified by concordant morphological and mtDNA traits. It seems to be reproductively isolated and apparently does not hybridize with sympatric brown hares. Phylogenetic analyses suggest that Italian and brown hares are not closely related sister taxa, but belong to distinct evolutionary lineages that dispersed in western Europe in different periods during the early Pleistocene. The Italian hare probably differentiated in isolated refuges in southern Italy during the last glaciation. Comparative analyses of mismatch distributions suggest that hares have had different demographic histories during the Pleistocene, which resulted in phylogeographical structuring in Italian hares, but not in brown and mountain (L. timidus) hares. The Italian hare is an endangered endemism and needs urgent conservation efforts.
Acetone preservation: a practical technique for molecular analysis
Authors: Fukatsu Takema
Published: March 2003

In attempts to establish a convenient and reliable method for field collection and archival preservation of insects and their endosymbiotic microorganisms for molecular analysis, acetone, ethanol, and other organic solvents were tested for DNA preservability of the pea aphid Acyrthosiphon pisum and its intracellular symbiotic bacterium Buchnera sp. After 6 months’ storage, not only the band of high‐molecular‐size DNA but also the bands of rRNA were well preserved in acetone, ethanol, 2‐propanol, diethyl ether and ethyl acetate. Polymerase chain reaction (PCR) assays confirmed that the DNA of both the insects and their symbionts was well preserved in these solvents. In contrast, methanol and chloroform showed poor DNA preservability. When water‐containing series of acetone and ethanol were examined for DNA preservability, acetone was apparently more robust against water contamination than ethanol. Considering that most biological materials contain high amounts of water, acetone may be a more recommendable preservative for DNA analysis than ethanol which has been widely used for this purpose. The DNA of various insects could be preserved in acetone at room temperature in good condition for several years. In addition to the DNA of the host insects, the DNA of their endosymbionts, including Buchnera and other mycetocyte symbionts, Wolbachia, and gut bacteria, was amplified by PCR after several years of acetone storage. The RNA and protein of the pea aphid and its endosymbiont were also preserved for several years in acetone. After 2 years’ storage in acetone, proteins of A. pisum could be analysed by sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS–PAGE) and immunoblotting, and the endosymbiotic bacteria were successfully detected by immunohistochemistry and in situ hybridization on the tissue sections.
A microsatellite survey of cattle from a centre of origin: the Near East
Authors: Loftus R. T. , Ertugrul O. , Harba A. H. , El‐Barody M. A. A. , MacHugh D. E. , Park S. D. E. , Bradley D. G.
Published: March 2003

Eight humpless cattle breeds from the Near East, three from Europe, one from West Africa and two zebu breeds from India were screened with 20 microsatellite loci. Breeds from the Near East revealed considerable levels of introgression from zebu cattle, which was apparent most in populations from the East and which declined in populations further West. This nonrandom pattern is suggestive of the introduction of zebu cattle from the East. Notwithstanding the overlay of zebu alleles, it was possible to demonstrate that Near Eastern cattle exhibited significantly higher levels of allelic diversity than breeds from other regions, which is consistent with the view that this region represents a primary domestication centre for Bos taurus cattle. The hypothesis that B. taurus and B. indicus cattle have separate domestic origins is also supported by the survey, a large genetic divergence being apparent between the nonhybrid taurine and zebu groups.
Microsatellite diversity and conservation of a relic trout population: McCloud River redband trout
Authors: Nielsen J. L. , Crow K. D. , Fountain M. C.
Published: March 2003

Rainbow trout native to the McCloud River, California, USA (Oncorhynchus mykiss stonei) are thought to represent a relic, nonanadromous trout adapted to harsh, fragmented environments. These fish, commonly named McCloud River ‘redband’ trout, survive in their most primitive form in a small, spring‐fed stream, Sheepheaven Creek, in the upper McCloud River drainage. Turn‐of‐the‐century fisheries records document both coastal anadromous steelhead and freshwater resident trout within the McCloud River drainage. The phylogenetic position of the McCloud River redband trout within O. mykiss has been debated for over 50 years. Based on phenotypic evidence, these fish were first reported as ‘southern Sierra golden trout’ by Wales in 1939.
Behnke (1970) considered them a relic subspecies of nonanadromous, fine‐scaled trout. Allozyme and mitochondrial DNA evidence suggested a coastal lineage. In this study, we examined within‐ and among‐basin genetic associations for Sheepheaven Creek redband trout using 11 microsatellite loci. Within‐basin analyses supported unique genetic characteristics in Sheepheaven Creek’s trout in comparisons with other McCloud River rainbow trout. Microsatellite data supported significant independence between Sheepheaven Creek fish and hatchery rainbow trout. Inter‐basin genetic distance analyses positioned Sheepheaven Creek fish with samples collected from Lassen Creek, a geographically proximate stream containing inland redband trout. California’s redband trout shared a close genetic association with Little Kern River golden trout (O.m. whitei) and isolated rainbow trout from Rio Santo Domingo, Baja, Mexico (O.m. nelsoni), suggesting a vicariant distribution of microsatellite diversity throughout the southern range of this species.
Mitochondrial DNA phylogeography and population history of the grey wolfCanis lupus
Authors: Vilà C. , Amorim I. R. , Leonard J. A. , Posada D. , Castroviejo J. , Petrucci‐Fonseca F. , Crandall K. A. , Ellegren H. , Wayne R. K.
Published: March 2003

The grey wolf (Canis lupus) and coyote (C. latrans) are highly mobile carnivores that disperse over great distances in search of territories and mates. Previous genetic studies have shown little geographical structure in either species. However, population genetic structure is also influenced by past isolation events and population fluctuations during glacial periods. In this study, control region sequence data from a worldwide sample of grey wolves and a more limited sample of coyotes were analysed. The results suggest that fluctuating population sizes during the late Pleistocene have left a genetic signature on levels of variation in both species. Genealogical measures of nucleotide diversity suggest that historical population sizes were much larger in both species and grey wolves were more numerous than coyotes. Currently, about 300 000 wolves and 7 million coyotes exist. In grey wolves, genetic diversity is greater than that predicted from census population size, reflecting recent historical population declines. By contrast, nucleotide diversity in coyotes is smaller than that predicted by census population size, reflecting a recent population expansion following the extirpation of wolves from much of North America. Both species show little partitioning of haplotypes on continental or regional scales. However, a statistical parsimony analysis indicates local genetic structure that suggests recent restricted gene flow.
Genetic relationships in the peregrine falcon (Falco peregrinus) analysed by microsatellite DNA markers
Authors: Nesje M. , Røed K. H. , Lifjeld J. T. , Lindberg P. , Steen O. F.
Published: March 2003

Microsatellite DNA markers were developed from a peregrine falcon (Falco peregrinus) and genetic relationships among peregrine falcons in southern Norway were analysed using the markers. The genomic DNA library was screened for the presence of dinucleotide microsatellite repeats. Twelve loci revealed polymorphism through the initial analysis of 24 unrelated peregrine falcons, and Mendelian inheritance was confirmed in two peregrine falcon families bred in captivity. The estimated mean probability of identical genotypes in two unrelated individuals was 3 × 10−8, and the combined exclusion probability for parentage testing was 0.99 and 0.94 for one or both parents unknown, respectively. The markers were used to investigate the parentage of peregrine broods from the same nest site from different breeding seasons, and subsequently the nest‐site fidelity of the breeding peregrines. High nest‐site fidelity was found by studying pairwise comparisons of relatedness (rxy) estimates among chicks at six nest sites from three different breeding seasons. Cross‐species amplifications showed that most loci also appeared to amplify polymorphic products in the gyrfalcon (F. rusticolus), merlin (F. columbarius), hobby (F. subbuteo) and kestrel (F. tinnunculus), demonstrating that the loci will provide powerful genetic markers in these falcons too.
Glacial refugia of limber pine (Pinus flexilisJames) inferred from the population structure of mitochondrial DNA
Authors: Mitton J. B. , Kreiser B. R. , Latta R. G.
Published: March 2003

To make inferences about the glacial refugia that harboured the limber pine,Pinus flexilisJames, we examined the range‐wide population structure of mitochondrial DNA (mtDNA) with eight size variants in the second intron ofnad1. The data consisted of haplotypes from 704 trees collected from 40 localities. The value ofFSTfor these populations was 0.80, which is a much larger value than has been reported for allozymes and chloroplast DNA (cpDNA) in limber pine, and it suggests that the number of seeds moving among localities per generation is ≈ 0.12. Gene flow of this magnitude would allow mutation and subsequent genetic drift to have a substantial impact on the population structure of mtDNA. The majority of the mtDNA haplotypes are restricted to minor portions of the geographical range. The data are consistent with mtDNA differentiation in seven glacial refugia, followed by dispersal out of those refugia.
Geographic distribution and frequency of a taurine Bos taurus and an indicine Bos indicus Y specific allele amongst sub‐Saharan African cattle breeds
Authors: Hanotte O. , Tawah C. L. , Bradley D. G. , Okomo M. , Verjee Y. , Ochieng J. , Rege J. E. O.
Published: March 2003

We report for the first time, and for the whole of sub‐Saharan Africa, the geographical distribution and the frequency of an indicine and a taurine Y specific allele amongst African cattle breeds. A total of 984 males from 69 indigenous African populations from 22 countries were analysed at the microsatellite locus INRA 124. The taurine allele is probably the oldest one on the continent. However, the taurine and the indicine alleles were present in 291 males (30%), and 693 males (70%), respectively. More particularly, 96% of zebu males (n = 470), 50% of taurine males (n = 263), 29% of sanga males (crossbreed Bos taurus × Bos indicus, n = 263) and 95% of zebu × sanga crossbred males (n = 56) had the indicine allele. The Borgou, a breed classified as zebu × taurine cross showed only the zebu allele (n = 12). The indicine allele dominates today in the Abyssinian region, a large part of the Lake Victoria region and the sahelian belt of West Africa. All the sanga males (n = 64) but only one from the Abyssinian region had the indicine allele. The taurine allele is the commonest only among the sanga breeds of the southern African region and the trypanotolerant taurine breeds of West Africa. In West Africa and in the southern Africa regions, zones of introgression were detected with breeds showing both Y chromosome alleles. Our data also reveal a pattern of male zebu introgression in Mozambique and Zimbabwe, probably originating from the Mozambique coast. The sanga cattle from the Lake Victoria region and the Kuri cattle of Lake Chad, cattle populations surrounded by zebu breeds were, surprisingly, completely devoid of the indicine allele. Human migration, phenotypic preferences by the pastoralists, adaptation to specific habitats and to specific diseases are the main factors explaining the present‐day distribution of the alleles in sub‐Saharan Africa.
Do discrepancies between microsatellite and allozyme variation reveal differential selection between sea and lagoon in the sea bass (Dicentrarchus labrax)?
Authors: Lemaire C. , Allegrucci G. , Naciri M. , Bahri‐Sfar L. , Kara H. , Bonhomme F.
Published: March 2003

In the present study the genetic structure of Dicentrarchus labrax (14 samples from the Mediterranean) was analysed at six microsatellite loci, in order to test the hypothesis that some enzymatic loci undergo selection between marine and lagoon habitat. Eight of the 14 samples were analysed at both microsatellite and allozyme markers. The analysis of the genetic variation among the Mediterranean samples showed that (i) FST values obtained with the six microsatellite loci were much smaller than those obtained with the 28 allozymes and (ii) microsatellite loci seemed to reflect more the geographical proximity than an ecological one. Thirteen enzymatic loci exhibited moderate to high values compared with microsatellites. This was interpreted as evidence that these allozymes are non‐neutral. However, only six loci seemed to be implicated in differentiation between marine and lagoon samples, the causes of selection being unknown for the others. A possible scenario of population dynamics of the sea bass between marine and lagoon habitat is suggested.