DNA metabarcoding provides great potential for species identification in complex samples such as food supplements and traditional medicines. Such a method would aid Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES) enforcement officers to combat wildlife crime by preventing illegal trade of endangered plant and animal species. The objective of this research was to develop a multi-locus DNA metabarcoding method for forensic wildlife species identification and to evaluate the applicability and reproducibility of this approach across different laboratories. A DNA metabarcoding method was developed that makes use of 12 DNA barcode markers that have demonstrated universal applicability across a wide range of plant and animal taxa and that facilitate the identification of species in samples containing degraded DNA. The DNA metabarcoding method was developed based on Illumina MiSeq amplicon sequencing of well-defined experimental mixtures, for which a bioinformatics pipeline with user-friendly web-interface was developed. The performance of the DNA metabarcoding method was assessed in an international validation trial by 16 laboratories, in which the method was found to be highly reproducible and sensitive enough to identify species present in a mixture at 1% dry weight content. The advanced multi-locus DNA metabarcoding method assessed in this study provides reliable and detailed data on the composition of complex food products, including information on the presence of CITES-listed species. The method can provide improved resolution for species identification, while verifying species with multiple DNA barcodes contributes to an enhanced quality assurance.

• Klíčová slova
• CITES, COI, Customs agencies, DNA metabarcoding, Endangered species, Traditional medicines, cyt b, matK, mini-barcodes, rbcL,
• MeSH
• DNA rostlinná genetika   MeSH
• ohrožené druhy * MeSH
• reprodukovatelnost výsledků MeSH
• rostliny klasifikace   genetika   MeSH
• taxonomické DNA čárové kódování * MeSH
• výpočetní biologie MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• DNA rostlinná MeSH

Reintroduction of captive-bred individuals into the wild is an important conservation activity. However, environmental conditions can influence developmental programming, potentially causing metabolic disorders in adults. These effects are investigated here for the first time in an endangered species. Using body weight and feed intake data for Iberian lynx (Lynx pardinus) (n = 22), we compared the growth of captive versus wild born and/or reared individuals. Captive-born individuals gained weight as a function of calorie intake, unlike wild-born individuals. When compared with females reared in the wild, captive-reared females achieved a larger body size, without evidence of obesity. Captivity-associated changes to metabolic programming may compromise survival in the wild if an increased body size incurs a greater energy requirement. Large body size may also confer a competitive advantage over smaller, wild-born individuals, disrupting the social organisation of existing wild populations, and inferring long-term implications for the phenotypic composition of wild populations.

• MeSH
• chování zvířat MeSH
• energetický příjem MeSH
• Lynx růst a vývoj   metabolismus   psychologie   MeSH
• ohrožené druhy MeSH
• sociální chování MeSH
• sociální izolace MeSH
• tělesná hmotnost MeSH
• zachování přírodních zdrojů MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Several steps of sturgeon somatic cell nuclear transfer (SCNT) have been recently established, but improvements are needed to make it a feasible tool to preserve the natural populations of this group of endangered species. The donor cell position inside the recipient egg seems to be crucial for its reprogramming; therefore by injecting multiple donor somatic cells instead of a single cell with a single manipulation, we increased the potential for embryo development. Using the Russian sturgeon Acipenser gueldenstaedtii as a multiple cell donor and sterlet Acipenser ruthenus as the non-enucleated egg recipient, we obtained higher proportion of eggs developing into embryos than previously reported with single-SCNT. Molecular data showed the production of a specimen (0.8%) contained only the donor genome with no contribution from the recipient, while two specimens (1.6%) showed both recipient and donor genome. These findings are the first report of donor DNA integration into a sturgeon embryo after interspecific cloning. In all, we provide evidence that cloning with the multiple donor somatic cells can be feasible in the future. Despite the fact that the sturgeon cloning faces limitations, to date it is the most promising technique for their preservation.

• MeSH
• embryonální vývoj * MeSH
• genom * MeSH
• klonování DNA metody   MeSH
• ohrožené druhy statistika a číselné údaje   MeSH
• ryby embryologie   genetika   MeSH
• techniky jaderného přenosu * MeSH
• zachování přírodních zdrojů metody   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Ice ages within Europe forced many species to retreat to refugia, of which three major biogeographic basic types can be distinguished: "Mediterranean", "Continental" and "Alpine / Arctic" species. However, this classification often fails to explain the complex phylogeography of European species with a wide range of latitudinal and altitudinal distribution. Hence, we tested for the possibility that all three mentioned faunal elements are represented within one species. Our data was obtained by scoring 1,307 Euphydryas aurinia individuals (46 European locations) for 17 allozyme loci, and sequencing a subset of 492 individuals (21 sites) for a 626 base pairs COI fragment. Genetic diversity indices, F statistics, hierarchical analyses of molecular variance, individual-based clustering, and networks were used to explore the phylogeographic patterns. The COI fragment represented 18 haplotypes showing a strong geographic structure. All but one allozyme loci analysed were polymorphic with a mean FST of 0.20, supporting a pronounced among population structure. Interpretation of both genetic marker systems, using several analytical tools, calls for the recognition of twelve genetic groups. These analyses consistently distinguished different groups in Iberia (2), Italy, Provence, Alps (3), Slovenia, Carpathian Basin, the lowlands of West and Central Europe as well as Estonia, often with considerable additional substructures. The genetic data strongly support the hypothesis that E. aurinia survived the last glaciation in Mediterranean, extra-Mediterranean and perialpine refugia. It is thus a rare example of a model organism that combines attributes of faunal elements from all three of these sources. The observed differences between allozymes and mtDNA most likely result from recent introgression of mtDNA into nuclear allozyme groups. Our results indicate discrepancies with the morphologically-based subspecies models, underlining the need to revise the current taxonomy.

• MeSH
• fylogeneze MeSH
• fylogeografie MeSH
• genetická variace * MeSH
• haplotypy MeSH
• mitochondriální DNA genetika   MeSH
• motýli genetika   MeSH
• ohrožené druhy * MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Evropa MeSH
• Názvy látek
• mitochondriální DNA MeSH

Assessing genetic diversity within populations of rare species and understanding its determinants are crucial for effective species protection. While a lot is known about the relationships between genetic diversity, fitness, and current population size, very few studies explored the effects of past population size. Knowledge of past population size may, however, improve our ability to predict future population fates. We studied Gentianella praecox subsp. bohemica, a biennial species with extensive seed bank. We tested the effect of current, past minimal and maximal population size, and harmonic mean of population sizes within the last 15 years on genetic diversity and fitness. Maximum population size over the last 15 years was the best predictor of expected heterozygosity of the populations and was significantly related to current population size and management. Plant fitness was significantly related to current as well as maximum population size and expected heterozygosity. The results suggested that information on past population size may improve our understanding of contemporary genetic diversity across populations. They demonstrated that despite the strong fluctuations in population size, large reductions in population size do not result in immediate loss of genetic diversity and reduction of fitness within the populations. This is likely due to the seed bank of the species serving as reservoir of the genetic diversity of the populations. From a conservation point of view, this suggests that the restoration of small populations of short-lived species with permanent seed bank is possible as these populations may still be genetically diverse.

• Klíčová slova
• Bottleneck effect, Expected heterozygosity, Microsatellites, Seed bank longevity, Storage effect,
• MeSH
• genetická variace MeSH
• hustota populace MeSH
• ohrožené druhy * MeSH
• populační genetika * MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Pulsatilla patens s.s. is a one of the most endangered plant species in Europe. The present range of this species in Europe is highly fragmented and the size of the populations has been dramatically reduced in the past 50 years. The rapid disappearance of P. patens localities in Europe has prompted the European Commission to initiate active protection of this critically endangered species. The aim of this study was to estimate the degree and distribution of genetic diversity within European populations of this endangered species. We screened 29 populations of P. patens using a set of six microsatellite primers. The results of our study indicate that the analyzed populations are characterized by low levels of genetic diversity (Ho = 0.005) and very high levels of inbreeding (FIS = 0.90). These results suggest that genetic erosion could be partially responsible for the lower fitness in smaller populations of this species. Private allelic richness was very low, being as low as 0.00 for most populations. Average genetic diversity over loci and mean number of alleles in P. patens populations were significantly correlated with population size, suggesting severe genetic drift. The results of AMOVA point to higher levels of variation within populations than between populations.The results of Structure and PCoA analyses suggest that the genetic structure of the studied P. patens populations fall into three clusters corresponding to geographical regions. The most isolated populations (mostly from Romania) formed a separate group with a homogeneous gene pool located at the southern, steppic part of the distribution range. Baltic, mostly Polish, populations fall into two genetic groups which were not fully compatible with their geographic distribution.Our results indicate the serious genetic depauperation of P. patens in the western part of its range, even hinting at an ongoing extinction vortex. Therefore, special conservation attention is required to maintain the populations of this highly endangered species of European Community interest.

• MeSH
• frekvence genu genetika   MeSH
• genetická variace genetika   MeSH
• genetický drift MeSH
• hustota populace MeSH
• mikrosatelitní repetice genetika   MeSH
• ohrožené druhy * MeSH
• populační genetika * MeSH
• Pulsatilla genetika   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• východní Evropa MeSH

The mega river ecosystem of the Yangtze River was once home to diverse aquatic megafauna but is increasingly affected by various anthropogenic stressors that have resulted in continuous loss of biodiversity, such as the probable extinction of Yangtze River Dolphin. The Chinese paddlefish, Psephurus gladius, was one of only two extant members of a relict lineage that was most diverse and widespread 34-75 million years ago. It is also one of the largest freshwater fish species, reaching up to 7 m in length. The Chinese paddlefish was once common in the Yangtze River, with c.25 t being harvested per annum during the 1970s. Populations have, however, declined drastically since the late 1970s as a result of overfishing and habitat fragmentation. Here, a basin-wide capture survey during 2017-2018 found 332 fish species, but did not find a single specimen of Chinese paddlefish. Furthermore, 140 historically reported fish species have not been found and most of them are considered highly endangered. Based on 210 sightings of Chinese paddlefish during the period 1981-2003, we estimated the timing of extinction to be by 2005, and no later than by 2010. In addition, the paddlefish probably became functionally extinct (i.e. it was unable to reproduce) by 1993, before it went extinct. It is likely that the lack of reproduction was among the major causes of extinction. As no individuals exist in captivity, and no living tissues are conserved for potential resurrection, the fish should be considered extinct according to the IUCN Red List criteria. The delayed extinction of Chinese paddlefish resulted from multiple threats, suggesting that optimizing conservation efforts on endangered Yangtze fauna is urgently needed.

• Klíčová slova
• Chinese paddlefish, Endangered fish, Extinction debt, Functional extinction, Sighting record, Yangtze River,
• MeSH
• biodiverzita MeSH
• extinkce biologická MeSH
• ohrožené druhy MeSH
• rybářství MeSH
• ryby MeSH
• sladká voda MeSH
• zachování přírodních zdrojů * MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Achorovermis testisinuosus gen. et sp. n. (Digenea: Aporocotylidae) infects the heart of the smalltooth sawfish, Pristis pectinata Latham (Rhinopristiformes: Pristidae), in the eastern Gulf of Mexico. Specimens of the new genus, along with the other blood flukes that infect batoids are similar by having an inverse U-shaped intestine and a curving testis as well as by lacking tegumental spines. The new genus differs from all of the other blood flukes infecting batoids by having an elongate body (>50 × longer than wide), a testis having >100 curves, and an ovary wholly anterior to the uterus. It differs from Ogawaia glaucostegi Cutmore, Cribb et Yong, 2018, the only other blood fluke infecting a rhinopristiform, by having a body that is >50 × (vs <30 ×) longer than wide, a testis that is >75 × (vs <40 ×) longer than wide and has >100 (vs <70) curves, an ovary wholly anterior to (vs lateral and dorsal to) the seminal vesicle, a uterus wholly posterior to (vs overlapping and lateral to both) the testis and ovary, and a sinuous (vs convoluted) uterus. The new species joins a small group of chondrichthyan blood flukes that lack tegumental spines: O. glaucostegi, Orchispirium heterovitellatum Madhavi et Rao, 1970, Myliobaticola richardheardi Bullard et Jensen, 2008, Electrovermis zappum Warren et Bullard, 2019. Blood flukes infecting batoids are further unique by having a curving testis. That is, the blood flukes infecting species within Selachii are morphologically distinct from those infecting species within the Batoidea (excluding Gymnurahemecus bulbosus Warren et Bullard, 2019). Based on the morphological similarity, we suspect that the new species shares a recent common ancestor with O. glaucostegi. The discovery of the new species brings the total number of chondrichthyan blood flukes to 11 species assigned to nine genera.

• Klíčová slova
• Chondrichthyes, Elasmobranchii, Endangered Species Act, North America, systematics, taxonomy,
• MeSH
• infekce červy třídy Trematoda epidemiologie   parazitologie   veterinární   MeSH
• nemoci ryb epidemiologie   parazitologie   MeSH
• ohrožené druhy MeSH
• prevalence MeSH
• rejnokovití * MeSH
• Trematoda klasifikace   fyziologie   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Mexický záliv epidemiologie   MeSH

INTRODUCTION: Sex is a fundamental characteristic of an individual. It is therefore puzzling why in some systems sex is precisely determined by a genotype, while in others it is influenced by the environment or even subtle, not to say random, factors. Some stochasticity in sex determination would be expected if environmental conditions did not have a large sex-specific effect on fitness. Although data are only available for a small fraction of species, geckos show exceptional variability in sex determination. METHODS: We tested the effects of three incubation temperatures on sex ratio and adult body size in the invasive gecko Phelsuma laticauda and the vulnerable gecko Phelsuma nigristriata. RESULTS: We document temperature-dependent sex determination (TSD) in both species. Only females hatched at a low temperature (24°C), whereas male production peaked at an intermediate temperature (28°C) and declined, at least in P. laticauda, again at the highest temperature (31°C). Interestingly, full siblings hatched from eggs glued together during the incubation at temperatures producing both sexes are often of the opposite sex. We found no significant effect of incubation temperature on adult body length. CONCLUSIONS: Documentation of TSD in the day geckos has implications for conservation practice in environmental management of endangered species or eradication of invasive species. However, it appears that a very subtle (random?) factor may also be involved in their sex determination. In line with this, we found no significant effect of incubation temperature on adult body length.

• Klíčová slova
• Environmental sex determination, Invasive species, Reptiles, Temperature,
• MeSH
• ještěři * fyziologie   genetika   MeSH
• ohrožené druhy * MeSH
• poměr pohlaví MeSH
• procesy určující pohlaví * MeSH
• teplota MeSH
• velikost těla MeSH
• zavlečené druhy MeSH
• životní prostředí * MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

The current state-of-the-art creates a sound basis for the preservation of unique species Cupressus dupreziana provided that targeted effort and care is devoted to the accomplishment of multiplication protocols. This review is to summarize known data on Cupressus dupreziana-specific characteristics, including abiotic stress resistance, and natural reproduction, and estimates the possibilities of ex situ conservation with an emphasis on the cultivation in vitro of this endangered species. As there is only limited information about the cultivation of C. dupreziana in vitro, we have included relevant data on the related species Cupressus sempervirens, where micropropagation techniques are well established, along with other information on species with similar fates and life strategies-Saharan olive Olea europaea subsp. laperrinei and myrtle Myrthus nivellei. The aim of this work is to enhance general understanding and to promote an interest in this relict plant species to contribute to more wide-ranging studies and to increase its chance of preservation. Besides others, reducing the number of species threatened with extinction is an essential and immediate task as high genetic variability of ecosystems is crucial for their stability under changing climatic conditions.

• Klíčová slova
• Cultivation in vitro, Duprez´cypress, Micropropagation, Olea europaea subsp. laperrinei, Saharan relict plant species,
• MeSH
• cypřiš (rod) růst a vývoj   fyziologie   MeSH
• ekosystém MeSH
• ohrožené druhy * MeSH
• rozmnožování fyziologie   MeSH
• šlechtění rostlin MeSH
• zeměpis MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH
• Geografické názvy
• severní Afrika MeSH