Trophic flexibility of the otter (Lutra lutra) in southern Italy

Diet composition of otters (Lutra lutra) was investigated in 2001 by spraints analysis (N=1323) on five rivers of southern Italy, with the aim of assessing the influence of fish availability, elevation and discharge on the consumption of food resources alternative to fish. Data were expressed as per cent frequency of occurrence (%FO) and per cent volume (%V). The study confirmed the great feeding adaptability of the otter that, in rivers partially interconnected and flowing on a small area, showed a strong fish eating habit in some rivers (Sinni and Mercure-Lao) and a diet mainly constituted by alternative resources in other ones (amphibians in the rivers Cogliandrino and Frido, crustaceans in the River Battendiero). Fish consumption for the five rivers was significantly correlated with fish biomass and with mean summer discharge, while it was inversely correlated with the mean altitude of the five rivers. The lack of a clear seasonality in the consumption of food sources alternative to fish together with the correlation between fish use and fish biomass for each river indicated fish availability as the main factor affecting otter relying to non-fish preys. Otter diet seemed influenced by the characteristics of river habitats (altitude, discharge and consequently fish biomass) more than by summer drought, typical of Mediterranean regions. The %FO and the %V allowed to drawn a similar picture of otter diet. Nonetheless the %V was useful for better illustrating diet variation among the different rivers and we argue that it could be useful in habitats where the otter feeds on preys with different proportions of indigestible remains.     

Relative spraint density and genetic structure of otter (Lutra lutra) along the Drava River in Hungary

In this study, we used genetic-based approaches to estimate population size and structure of Eurasian otter along the Drava River in Hungary, and compared these results to traditional survey-based methods. The relative spraint density of otter was estimated based on the number of fresh (D_f) and total number (D_t) of spraints collected on standard routes over a 2-year period. Nine microsatellite loci were screened, generating 17 individual otter genotypes composed of 45 different alleles. The expected heterozygosity ranged from 0.53 to 0.89 and observed heterozygosity from 0.25 to 0.92. The mean density (D_g) estimated over six different sites was 0.17 individuals per km of shoreline. A close correlation was found between the number of genotypes and spraint counts along a standard route (fresh spraints: r_P=0.85, P<0.01; total spraints r_P=0.76, P<0.05). All genotypes found within the 50 km-long study area were closely related (D_m ranged between 0.08 and 0.21).     

Isolation and characterization of tetramicrosatellite DNA markers in the Eurasian otter (Lutra lutra)

Eight new tetramicrosatellite loci for Eurasian otter (Lutra lutra) were designed. Polymorphism of these eight loci in 29 otter individuals was tested. The results indicated that the allele numbers of each of loci ranged from three to five and the observed heterozygosity from 0.483 to 0.828. These new loci can be useful for population genetic research on otters and help improve the resolution of individual identification using noninvasive method.     

Genetic structure of Eurasian otter (Lutra lutra,Carnivora: Mustelidae) populations from the western Baltic sea region and its implications for the recolonization of north‐western Germany

We analysed 229 Eurasian otters from eastern and north‐western Germany, Denmark and southern Sweden based on sequences of the mtDNA control region and genotypes at 12 polymorphic nuclear microsatellite loci. The main focus of the study lay on the north‐west German otters from Schleswig‐Holstein, which represent a newly established and expanding population recolonizing a formerly inhabited region, thereby closing the distributional gap between the large eastern population and the long‐isolated and genetically depleted Danish population. As found before in this species, mtDNA variability was very low but we identified a hitherto unknown haplotype in Sweden. Expected heterozygosities were between 0.46 and 0.69 and thus within the range known from the literature. There were only weak signs of a founder effect in the Schleswig‐Holstein otters with respect to allelic diversity (but not heterozygosity). Nevertheless, there was a statistically significant bottleneck signal based on deviations from mutation‐drift equilibrium in Schleswig‐Holstein. All statistical approaches (amova s, factorial correspondence analysis, assignment tests and Bayesian structure analysis) unequivocally showed that north‐western Germany has so far been predominantly recolonized by otters of east German origin, but we also found evidence of admixture through immigration from Denmark. Both the Danish and the north‐west German otter population will benefit from further exchange in the wake of the ongoing range expansion.     

Sex identification of Eurasian otter (<Emphasis Type="Italic">Lutra lutra</Emphasis>) non-invasive DNA samples using ZFX/ZFY sequences

We developed a new PCR/RFLP system targeted to amplify and cut a segment of the ZFX/ZFY gene in non-invasive otter (Lutra lutra) samples. This assay produced one sex-specific fragment in females (XX genotypes) and two fragments in males (XY genotypes), and is intrinsically more reliable than alternative systems (e.g., SRY genes) that are based on the amplification of a single Y-linked fragment. The new primer pair correctly identified the sex of 23 DNA extracted from sexed otter tissues. This procedure was used to sex unknown DNA extracted from otter scats. A multi-tube approach led to identify the sex of 72/91 (79%) samples, using a minimum of two PCR replicates. This procedure is currently used in non-invasive genetic monitoring of Italian otter populations.     

Source and sink dynamics of density-dependent otter (<Emphasis Type="Italic">Lutra lutra</Emphasis>) populations in rivers of central Finland

Long-term studies were carried out in central Finland between 1985 and 2003 to examine the temporal and spatial variation in the density of otter populations. Snow tracking was used to estimate the total population and the number of litters in the study area. In total 52 otters, including 16 cubs in 11 litters, lived in the study area (1,650 km²) in 2002–2003. The otter population clearly increased during the study period. The increase in density of the otter population was sigmoid, indicating that the population had reached the local carrying capacity. The density of the population was 0.12 individuals per river ha in 1985 and 0.29 individuals per river ha in 2002. The number of cubs per litter decreased when the density of the population increased. Density-dependent offspring production, together with the auto-correlation function of growth rate, indicates intraspecific competition in otter populations. Otters in a few river systems produced most of the cubs, creating several small source populations in the entire study area. Otters in secondary (sink) habitats had a low reproduction rate. Most otters lived in river systems with large lake surfaces. The number or area of lakes within the river system correlated positively with the total number of otters, litters and cubs in the river system. The six river systems (out of 16) with the largest water area of lakes produced 81.2% of all cubs born in the study area. However, the population growth rate per river hectare or per river kilometre was equal in all kinds of river systems. Thus, among local otter populations in central Finland, a source–sink system between different habitats seems to be prevalent.     

Estimating population size of endangered brush-tailed rock-wallaby (Petrogale penicillata) colonies using faecal DNA

The brush-tailed rock-wallaby (Petrogale penicillata) is an endangered species in southeastern Australia and many of the remaining populations are declining. The steep rocky habitat and shy nature of the species make it difficult to obtain data on population parameters such as abundance and recruitment. Faecal pellet counts from scat plots are commonly used to monitor population trends but these are imprecise and difficult to relate to absolute population size. We conducted a noninvasive genetic sampling 'mark-recapture' study over a 2-year period to identify individuals from faecal DNA samples and estimate the population size of four brush-tailed rock-wallaby colonies located in Wollemi National Park, New South Wales. Scat plots in rock-wallaby colonies were used as sample collection points for this study. Two separate population estimates were carried out for three of the colonies to determine if we could detect recruitment and changes in population size. We determined that there was one large colony of an estimated 67 individuals (95% confidence interval: 55-91) and three smaller colonies. Monitoring of the smaller colonies also detected possible population size increases in all three. Our results indicate that faecal DNA analysis may be a promising method for estimating and monitoring population trends in this species particularly when used with a traditional field survey method.     

DNA “fingerprinting” and the genetic management of a captive chimpanzee population (Pan troglodytes)

DNA fingerprinting probes are cloned sequences which simultaneously detect a large number of similar hypervariable loci in the target DNA. The resulting highly polymorphic pattern visualized on an autoradiograph allows resolution of questions concerning individual identification and parentage. M13 bacteriophage has been used as a DNA fingerprinting probe for paternity ascertainment among captive chimpanzees housed in multi-male groups as part of the National Chimpanzee Breeding and Research Program. In 31 cases of unknown paternity where DNA samples for mother, offspring, and all potential sires were available, DNA fingerprinting with M13 resulted in the unambiguous assignment of paternity for all 31 infants. Knowledge of pedigrees among the captive-born animals is used to address several issues important in the genetic management of captive breeding colonies, including estimation of effective population size and of the rate of decline in genetic variability, variance in male and female reproduction, and the effect of social dominance on male reproductive success. Our analysis demonstrates the beneficial effects of genetic management by comparing the managed dedicated cohort to the Bastrop colony as a whole.     

Selected urinary metal reference concentrations of the Viennese population – urinary metal reference values (Vienna)

Reference concentrations of metals are the basis for risk assessment studies and for estimation of type and magnitude of environmental and occupational exposure. Urine is often used as a specimen for monitoring studies, as it plays an important role in the elimination of various substances from the body and in addition it can be collected easily. Therefore, the urinary levels of seven trace elements (Al, Co, Cr, Mo, Nb, Ni, Ti) were determined by atomic spectrometric methods (ICP-MS and GFAAS) in 100 urine samples of the Viennese population. The obtained creatinine adjusted concentrations (medians) are in μg/g 6.1, 1.5, 1.1, 46.2, 0.4, 0.1, and 8.0 for Al, Co, Cr, Mo, Nb, Ni, and Ti, respectively.     

Divergence time uncertainty and historical biogeography reconstruction – an example from Urophylleae (Rubiaceae)

Aim When hypotheses of historical biogeography are evaluated, age estimates of individual nodes in a phylogeny often have a direct impact on what explanation is concluded to be most likely. Confidence intervals of estimated divergence times obtained in molecular dating analyses are usually very large, but the uncertainty is rarely incorporated in biogeographical analyses. The aim of this study is to use the group Urophylleae, which has a disjunct pantropical distribution, to explore how the uncertainty in estimated divergence times affects conclusions in biogeographical analysis. Two hypotheses are evaluated: (1) long‐distance dispersal from Africa to Asia and the Neotropics, and (2) a continuous distribution in the boreotropics, probably involving migration across the North Atlantic Land Bridge, followed by isolation in equatorial refugia. Location Tropical and subtropical Asia, tropical Africa, and central and southern tropical America. Methods This study uses parsimony and Bayesian phylogenetic analyses of chloroplast DNA and nuclear ribosomal DNA data from 56 ingroup species, beast molecular dating and a Bayesian approach to dispersal–vicariance analysis (Bayes‐DIVA) to reconstruct the ancestral area of the group, and the dispersal–extinction–cladogenesis method to test biogeographical hypotheses. Results When the two models of geographic range evolution were compared using the maximum likelihood (ML) tree with mean estimates of divergence times, boreotropical migration was indicated to be much more likely than long‐distance dispersal. Analyses of a large sample of dated phylogenies did, however, show that this result was not consistent. The age estimate of one specific node had a major impact on likelihood values and on which model performed best. The results show that boreotropical migration provides a slightly better explanation of the geographical distribution patterns of extant Urophylleae than long‐distance dispersal. Main conclusions This study shows that results from biogeographical analyses based on single phylogenetic trees, such as a ML or consensus tree, can be misleading, and that it may be very important to take the uncertainty in age estimates into account. Methods that account for the uncertainty in topology, branch lengths and estimated divergence times are not commonly used in biogeographical inference today but should definitely be preferred in order to avoid unwarranted conclusions.     

Cynanchum and the Cynanchinae (Apocynaceae – Asclepiadoideae): a molecular, anatomical and latex triterpenoid study

The phylogeny of the genus Cynanchum s. str. is studied using cpDNA spacers and ITS. Morphological, anatomical and latex triterpenoid data are interpreted in light of the molecular results, and discrepancies are discussed. Vegetative characters are better indicators of relationship than floral characters, especially corona characters. The monophyly of all Malagasy species and, nested within the latter, of all stem-succulent taxa is ascertained and the genera Folotsia, Karimbolea, Platykeleba and Sarcostemma are subsumed under Cynanchum. One African species, C. galgalense, is excluded from Cynanchum.     

Phylogeny of Caragana (Fabaceae) based on DNA sequence data from rbcL, trnS–trnG, and ITS

Phylogenetic relationships of 48 species of Caragana (Fabaceae: tribe Hedysareae) and one representative each of Astragalus, Calophaca, Halimodendron, and Hedysarum are estimated from DNA sequences of the rbcL gene, trnS–trnG intron and spacer, and ITS region. At least one representative of all five sections and 12 series within Caragana are included. Analyses yielded strongly supported clades corresponding to sections Caragana, Bracteolatae, and Frutescentes. The species of section Jubatae are distributed among three strongly supported clades, i.e., one with the species of section Bracteolatae, another with two species of section Spinosae, and a third as sister to section Frutescentes. All but the last of these six clades are corroborated by at least one unambiguously traced morphological character. The placement of the other four species of section Spinosae are not well supported and lack unambiguous morphological synapomorphies, and the samples of Calophaca and Halimodendron nest within Caragana with weak support.     

Microsatellite and mitochondrial DNA analyses of Atlantic bluefin tuna (Thunnus thynnus thynnus) population structure in the Mediterranean Sea

Genetic variation was surveyed at nine microsatellite loci and the mitochondrial control region (868 bp) to test for the presence of genetic stock structure in young-of-the-year Atlantic bluefin tuna (Thunnus thynnus thynnus) from the Mediterranean Sea. Bluefin tuna were sampled over a period of 5 years from the Balearic and Tyrrhenian seas in the western basin of the Mediterranean Sea, and from the southern Ionian Sea in the eastern basin of the Mediterranean Sea. Analyses of multilocus microsatellite genotypes and mitochondrial control region sequences revealed no significant heterogeneity among collections taken from the same location in different years; however, significant spatial genetic heterogeneity was observed across all samples for both microsatellite markers and mitochondrial control region sequences (FST=0.0023, P=0.038 and PhiST=0.0233, P=0.000, respectively). Significant genetic differentiation between the Tyrrhenian and Ionian collections was found for both microsatellite and mitochondrial markers (FST=0.0087, P=0.015 and PhiST=0.0367, P=0.030, respectively). These results suggest the possibility of a genetically discrete population in the eastern basin of the Mediterranean Sea.     

Evolution of Asian ‘lowland’ taxa in relation to the Alpine–Himalayan Tertiary orogenic belt – Insight from a cladistic analysis of Maladera (Cycloserica) (Coleoptera: Scarabaeidae: Sericini)

Among the south Asian Sericini only a very few endemic monophyletic lineages developed in the lowland areas, such as the taxa belonging to Maladera (subgenus Cycloserica). Twenty species of Sericini (Coleoptera: Scarabaeidae) representing the principal lineages of Maladera occurring in the Palearctic region were included in a parsimony analysis based on 37 morphological characters. Maladera (Cycloserica) was found be monophyletic, composed by a clade comprising (Maladera caspia (Cycloserica excisipes, Leucoserica arenicola)) being distributed in the Middle Asian lowlands and its sister group the Maladera quinquidens-group with four endemic species in the Himalayan and North Indian lowlands. The ranges of these two clades as well as of the sister group of Cycloserica reveal a separating effect by the uplifted Himalayan–Alpine belt. It is hypothesized that these two Cycloserica lineages existed before they dispersed into the two separated areas, the northern Indian subcontinent (Himalayas) and the Middle Asian lowland. Due to the sister group relationship of Cycloserica and Leucoserica, both nested deeply within the species of Maladera, it seems reasonable to consider both synonymous with Maladera. Based on this conclusion one new combination is necessary, Maladera (Cycloserica) arenicola, and Leucoserica Reitter, 1896, is placed in synonymy with Cycloserica Reitter, 1896).     

Urinary hormone analysis assists reproductive monitoring and sex identification of bell frogs (Litoria raniformis)

With the world currently facing a global amphibian extinction crisis, the development of techniques to help meet the needs of conservation managers and researchers studying the reproductive biology of amphibians is needed. Here, we developed enzyme immunoassays to measure estrone, testosterone, and progesterone hormone metabolites in the urine of Litoria raniformis, the southern bell frog. Concentrations of urinary estrone, testosterone, and progesterone increased during the breeding season for females (P < 0.05). Concentrations of urinary testosterone and progesterone increased for males during the breeding season compared with that for months where no reproductive behaviors were observed (P < 0.05). Furthermore, urinary estrone concentrations proved to be a reliable sexing tool for adult frogs, with no overlap between the sexes in 98% of cases, regardless of season. There was no difference in estrone (P = 0.204) or testosterone (P = 0.485) metabolite concentrations between samples taken immediately upon capture and those taken 12 to 24 h later from the same individual. Progesterone metabolite concentrations were lower on Day 2 than upon collection (P = 0.004). This is the first study to show that urinary hormone analysis can be a useful technique for reproductive monitoring in an amphibian. Additionally, hormone metabolite measures offer promise as sex identification tools for monomorphic species and for those whose secondary sex characteristics are visible only during the breeding season.     

异叶泽兰的遗传多样性和居群历史动态研究

异叶泽兰属于菊科泽兰属,是该属分布海拔相对较高的植物,分布在青藏高原东部和横断山海拔1700～3000 m的地区.该文利用ycf6-psbM和rpl32-trnL两个叶绿体DNA(cpDNA)片段以及核DNA片段ITS(nITS)作为分子标记,研究了异叶泽兰的遗传多样性及其分布特征,探讨了其居群历史动态.结果表明:(1...     

单花、有侧花牡丹品种花芽分化特点及内源激素变化

为比较单花牡丹品种‘梨花粉’、有侧花牡丹品种‘姊妹游春’和‘云鄂粉’的花芽分化进程、形态特征差异及花芽分化期内源激素的变化,该研究通过解剖和石蜡切片,观察‘梨花粉’‘姊妹游春’和‘云鄂粉’的花芽分化过程,采用酶联免疫吸附分析法测定不同分化阶段‘梨花粉’和‘姊妹游春’的生长素(IAA)、脱落酸(ABA)、赤霉素(GA₃)及玉米素核苷(ZR)的含量。结果表明：(1)‘梨花粉’花芽分化仅6个时期;有侧花品种‘姊妹游春’和‘云鄂粉’花芽分化为11个时期,顶花原基先分化,侧花原基后分化。(2)叶原基分化期至苞片原基分化期,‘姊妹游春’和‘梨花粉’花芽中ABA、GA₃及ZR的含量均升高,IAA含量降低;‘姊妹游春’侧花原基分化起始阶段,ABA、GA₃含量降低,ZR、IAA含量升高。(3)‘梨花粉’ABA/IAA比值在形态分化期时最低。综上认为,有侧花牡丹品种花芽分化比单花牡丹品种早,分化时间长,顶花和侧花部分花器官分化期重叠;较高水平的ZR、IAA以及较低的ABA、GA₃有利于启动侧花原基分化。该研究结果为牡丹...