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Screening the foods of an endangered parrot, the kakapo (Strigops habroptilus), for oestrogenic activity using a recombinant yeast bioassay.

In recent years the possibility of environmental oestrogens affecting the reproduction of vertebrates has become an issue of both public and scientific interest. Although the significance of such chemicals remains controversial there is clear evidence that, in some contexts, environmental oestrogens can influence the fertility of vertebrates. Highly endangered species represent a situation in which even modest reductions in the fertility of key individuals may have implications for the survival of the entire species. This paper reports the screening of both natural and supplementary foods of the kakapo (Strigops habroptilus), a critically endangered New Zealand nocturnal parrot, for oestrogenic activity using a recombinant yeast based bioassay. Low levels of oestrogenic activity were detected in one of the 'chick-raising' foods, but no oestrogenic activity was detected in the adult supplementary foods. The oestrogenicity of a range of phytochemicals possibly associated with the kakapo natural diet was also examined. Two such phytochemicals, podocarpic acid and its reduced derivative podocarpinol, showed weak oestrogenic activity (approximately 10(-6) and 10(-4) of the activity of 17-beta-oestradiol, respectively).

Abietanes↗

Erysipelas in the critically endangered kakapo (Strigops habroptilus).

This paper describes the effect of a disease outbreak on the success of a translocation for conservation management in a critically endangered species. Three juvenile kakapo from a group of 19 translocated birds died within 72 h of transport between New Zealand offshore islands. Clinical findings, gross necropsy changes, cytology, histopathology and bacterial culture confirmed systemic disease caused by Erysipelothrix rhusiopathiae. On the island from which the kakapo were sourced, positive cultures of E. rhusiopathiae were obtained from the medulla of the ulna from 10 out of 15 seabird carcasses examined, suggesting that this could be the source of infection for the kakapo. Immediately after the diagnosis, all of the translocated birds were re-captured and treated with antibiotics. A vaccination programme has commenced using a commercial killed bacterin developed for turkeys. The disease outbreak has had costly implications for the population and conservation management of the species. This is the first report of erysipelas in wild parrots, and the first report of the management of erysipelas in a critically endangered wild population of birds.

Animals↗

Genetic diversity and geographic differentiation in endangered Ammopiptanthus (Leguminosae) populations in desert regions of northwest China as revealed by ISSR analysis.

BACKGROUND AND AIMS: The desert legume genus Ammopiptanthus comprises two currently endangered species, A. mongolicus and A. nanus. Genetic variability and genetic differentiation between the two species and within each species were examined. METHODS: Inter-simple sequence repeat (ISSR) marker data were obtained and analysed with respect to genetic diversity, structure and gene flow. KEY RESULTS: Despite the morphological similarity between A. mongolicus and A. nanus, the two species are genetically distinct from each other, indicated by 63 % species-specific bands. Low genetic variability was detected for both population level (Shannon indices of diversity Hpop = 0.106, percentage of polymorphic loci P = 18.55 % for A. mongolicus; Hpop = 0.070, P = 12.24 % for A. nanus) and species level (Hsp = 0.1832, P = 39.39 % for A. mongolicus; Hsp = 0.1026, P = 25.89 % for A. nanus). Moderate genetic differentiation was found based on different measures (AMOVA PhiST and Hickory B) in both A. mongolicus (0.3743-0.3744) and A. nanus (0.2162-0.2369). CONCLUSIONS: The significant genetic difference between the two species might be due to a possible vicariant evolutionary event from a single common ancestor through the fragmentation of their common ancestor's range. Conservation strategies for these two endangered species are proposed.

Analysis of Variance↗

Allozyme variation in the endangered insular endemic Castilleja grisea.

BACKGROUND AND AIMS: Genetic diversity in Castilleja grisea, an endangered, perennial herb endemic to San Clemente Island, California was investigated. Subsequent to the elimination of goats from the island in 1992, many populations of C. grisea have reappeared and have been increasing in size. METHODS: Nineteen populations were surveyed for their genotype at 19 allozyme loci. KEY RESULTS: At the taxon level, 57.9 % of loci are polymorphic with A(P) = 3.09 and H(E) = 0.137. Populations averaged 33.0 % polymorphic loci with A(P) = 2.43 and H(E) = 0.099. Most variation is found within rather than among populations (G(ST) = 0.128), although differentiation among populations is significant. Genetic identities range from I = 0.960 to I = 1.000 with mean I = 0.990. There is no significant relationship between genetic and geographic distance. Gene flow among populations is Nm = 2.50 based on private alleles and Nm = 1.70 based on F(ST). Outcrossing rates based on fixation indices average t = 1.01, indicating a primarily out-crossed mating system. CONCLUSIONS: The observed genetic variation is moderately high, unusually so for an insular endemic species, suggesting that C. grisea may not have lost substantial genetic variation during 150 years of overgrazing, and indicating that it is unlikely to be endangered by genetic factors.

Acanthaceae↗

Genetic fingerprinting of germplasm accessions as an aid for species conservation: a case study with Borderea chouardii (Dioscoreaceae), one of the most critically endangered Iberian plants.

BACKGROUND AND AIMS: Molecular markers have changed previous expectations about germplasm collections of endangered plants, as new perspectives aim at holding a significant representation of all the genetic diversity in the studied species to accomplish further conservation initiatives successfully. Borderea chouardii is a critically endangered allotetraploid dioecious member of Dioscoreaceae, known from a single population in the Iberian pre-Pyrenees. This population was reported to be highly structured into two genetically distinct groups of individuals corresponding to their spatial separation along the vertical cliff where it grows. In 1999, the Spanish Government of Aragón launched the first conservation programme for the ex situ preservation of this species, and since then a seed collection has been conserved at the Germplasm Bank of the Universidad Politécnica de Madrid. However, as some seed samples had not been labelled clearly at the time of collection, their origin was uncertain. METHODS: Genetic variation in germplasm accessions of B. chouardii was investigated using microsatellite (simple sequence repeat; SSR) markers. KEY RESULTS: The 17 primer pairs used detected 62 SSR alleles in the 46 samples analysed from five different germplasm stocks. Eight alleles scored from the wild population were not detected in the germplasm samples analysed. The relatedness of the germplasm samples to the wild subpopulations through neighbour-joining clustering, principal coordinates analysis (PCO) and assignment tests revealed a biased higher representation of the genetic diversity of the lower cliff (43 samples) subpopulation than that of the upper cliff (three samples). CONCLUSIONS: The collection of additional samples from the upper cliff is recommended to achieve a better representation of the genetic diversity of this subpopulation. It is also recommended that these stocks should be managed separately according to their distinct microspatial origin in order to preserve the genetic substructuring of the wild population.

Alleles↗

Research in nondomestic species: experiences in reproductive physiology research for conservation of endangered felids.

Tremendous strides have been made in recent years to broaden our understanding of reproductive processes in nondomestic felid species and further our capacity to use this basic knowledge to control and manipulate reproduction of endangered cats. Much of that progress has culminated from detailed scientific studies conducted in nontraditional laboratory settings, frequently at collaborating zoological parks but also under more primitive conditions, including in the field. A mobile laboratory approach is described, which incorporates a diverse array of disciplines and research techniques. This approach has been extremely useful, especially for conducting gamete characterization and function studies as well as reproductive surveys, and for facilitating the development of assisted reproductive technology. With continuing advances in assisted reproduction in rare felids, more procedures are being conducted primarily as service-related activities, targeted to increase effectiveness of species propagation and population management. It can be a challenge for both investigators and institutional animal care and use committees (IACUCs) to differentiate these service-based procedures from traditional research studies (that require IACUC oversight). For research with rare cat species, multi-institutional collaboration frequently is necessary to gain access to scientifically meaningful numbers of study subjects. Similarly, for service-based efforts, the ability to perform reproductive procedures across institutions under nonstandard laboratory conditions is critical to applying reproductive sciences for managing and preserving threatened cat populations. Reproductive sciences can most effectively assist population management programs (e.g., Species Survival Plans) in addressing conservation priorities if these research and service-related procedures can be conducted "on the road" at distant national and international locales. This mobile laboratory approach has applications beyond endangered species research, notably for other scientific fields (e.g., studies of hereditary disease in domestic cat models) in which bringing the laboratory to the subject is of value.

Animals↗

Establishing a captive broodstock for the endangered bonytail chub (Gila elegans).

It is crucial for endangered species to retain as much genetic variation as possible to enhance recovery. Bonytail chub (Gila elegans) is one the most imperiled freshwater fish species, persisting as a declining population of large and old individuals primarily in Lake Mohave on the lower Colorado River. Establishment of a new captive broodstock from the 1981 F1 progeny of at most 10 wild fish plus any newly captured wild fish is evaluated and reviewed. The effective number of founders contributing to the 1981 F1 progeny appears quite small, varying from approximately 3.5, based on F1 allozyme data and supported by mtDNA data, to approximately 8.5, based on the original production records. Using a sample of these progeny to initiate a new broodstock further reduces the effective number of founders. With even the most optimistic evaluation of the amount of genetic variation in F1 progeny, it is obvious that including wild fish in the broodstock is essential to increase the amount of genetic variation. The approach given here could be applied to retain genetic variation in other endangered species in a captive broodstock until they have stable natural populations of adequate size.

Animals↗

Cross-species amplification of Bovidae microsatellites and low diversity of the endangered Korean goral.

The Korean goral (Nemorhaedus caudatus) is an endangered species of wild goat. The conservation and management of this species could benefit from a better understanding of its genetic diversity and structure. Cross-species amplification of 34 Bovidae microsatellite loci was tested on a panel of 6 Korean gorals and 10 domestic goats. After polymerase chain reaction (PCR) optimization, 29 (85.3%) microsatellite loci amplified successfully for the Korean gorals and 27 (79.4%) for the domestic goats. Of the amplified products, 16 (55.2%) were polymorphic in the Korean goral and 22 (81.5%) in domestic goats. Nei's unbiased mean heterozygosity and mean allele number per locus were, respectively, 0.356 and 2.6 in the Korean goral and 0.636 and 4.8 in domestic goats. Low genetic diversity in the Korean gorals observed in this preliminary microsatellite survey suggests an urgent need for further detailed study of genetic diversity in Korean goral populations and a population management strategy based on these studies. Current results of cross-species amplification of domestic Bovidae microsatellites could be employed for the necessary population genetic studies on the Korean goral and other endangered Caprinae species.

Animals↗

Acoustic detection and satellite-tracking leads to discovery of rare concentration of endangered North Pacific right whales.

The North Pacific right whale, Eubalaena japonica, is one of the most endangered species of whale in the world. On 10 August 2004, two right whales were located in the Bering Sea using headings to right whale calls provided by directional sonobuoys. A satellite-monitored radio tag attached to one of these whales functioned for 40 days. Over the 40-day period, this whale moved throughout a large part of the southeast Bering Sea shelf, including areas of the outer-shelf where right whales have not been seen in decades. In September, multiple right whales were acoustically located and subsequently sighted by another survey vessel approaching a near-real-time position from the tag. An analysis of photographs confirmed at least 17 individual whales (not including the tagged whales). Genetic analysis of biopsy samples identified 17 individuals: 10 males and 7 females. The discovery of seven females was significant, as only one female had been identified in the past. Genetics also confirmed the presence of at least two calves. Although the future of this population is highly uncertain, the discovery of additional females and calves gives some hope that this most critically endangered of all whale populations may still possess the capacity to recover.

Acoustics↗

Initial stages of reproductive isolation in two species of the endangered Sonoran topminnow.

Long-term geographic isolation can result in reproductive incompatibilities due to forces such as mutation, genetic drift, and differential selection. In the Sonoran topminnow, molecular genetic studies of mtDNA, microsatellites, and MHC genes have shown that the endangered Gila and Yaqui topminnows are substantially different, suggesting that divergence took place approximately two million years ago. Here we examined hybrid crosses and backcrosses between these two allopatric taxa to evaluate the accumulation of postmating barriers to reproduction. These results are then compared with results from a previous study where male topminnows were shown to mate assortatively with conspecific females. Despite their preference for conspecific mates, both types of interspecific crosses successfully produced offspring. There was evidence of reduced hybrid fitness, including smaller mean brood size and male-biased sex ratio, for some classes of backcrosses. Brood sizes and interbrood intervals varied significantly when hybrids were subdivided into different cross categories. Our results illustrate the importance of distinctly defining hybrid classes in studies of reproductive isolation. To our knowledge, this is the first such detailed evolutionary analysis in endangered fish taxa.

Animals↗

Patterns of variability and gene flow in Medicago citrina, an endangered endemic of islands in the western Mediterranean, as revealed by amplified fragment length polymorphism (AFLP).

Medicago citrina is an endangered western Mediterranean endemic that grows only on small islets of the Balearic archipelago and off the eastern Spanish coast. Only 10 isolated subpopulations are currently known (four from Ibiza, three from Cabrera, two from Columbretes and one from an offshore islet in northern Alicante province), constituting a severely fragmented genetic system. Data were analysed with the unweighted pair-group method using arithmetic averages (UPGMA) and principle coordinates analysis (PCOA), revealing several distinct groups. Genetic diversity indices indicated that Ibizan subpopulations had the highest genetic variability (Nei's index: 0.1463; Shannon's index: 0.228), whereas the lowest variability was found in Alicante (Nei's index: 0.035; Shannon's index: 0.050) and Cabrera (Nei's index: 0.068; Shannon's index: 0.104). These latter populations show the highest FST values (FST = 0.548) revealing high differentiation between them. Columbretes subpopulations formed a defined single group, although it also included some Ibizan samples. The smallest FST values, obtained between Ibiza and Columbretes (FST = 0.185), are not correlated with geographical proximity, but appear to be related to the geologically recent volcanic origin of the Columbretes islands (300,000 years ago). According to the distribution of the Ibizan samples in the dendrogram and the FST values, the best hypothesis is to regard the Ibizan subpopulations as the centre of genetic diversity of the currently known subpopulations. Our results suggest migratory scenarios from Ibiza to Columbretes based mainly on zoochory probably by seabirds. Finally, recommendations are provided for management strategies to facilitate the conservation of this endangered species.

Cluster Analysis↗

Conservation genetics and demographic history of the endangered Cape Fear shiner (Notropis mekistocholas).

We examined allelic variation at 22 nuclear-encoded markers (21 microsatellites and one anonymous locus) and mitochondrial (mt)DNA in two geographical samples of the endangered cyprinid fish Notropis mekistocholas (Cape Fear shiner). Genetic diversity was relatively high in comparison to other endangered vertebrates, and there was no evidence of small population effects despite the low abundance reported for the species. Significant heterogeneity (following Bonferroni correction) in allele distribution at three microsatellites and in haplotype distribution in mtDNA was detected between the two localities. This heterogeneity may be due to reduced gene flow caused by a dam built in the early 1900 s. Bayesian coalescent analysis of microsatellite variation indicated that effective population size of Cape Fear shiners has declined in recent times (11-25 435 years ago, with highest posterior probabilities between 126 and 2007 years ago) by one-two orders of magnitude, consistent with the observed decline in abundance of the species. A decline in effective size was not indicated by analysis of mtDNA, where sequence polymorphism appeared to carry the signature of an older expansion phase that dated to the Pleistocene ( approximately 12 700 > 1 million years ago). Cape Fear shiners thus appear to have undergone an expansion phase following a glacial cycle but to have declined significantly in more recent times. These results suggest that rapidly evolving markers such as microsatellites may constitute a suitable tool when inferring recent demographic dynamics of populations.

Animals↗

Phylogeography of the endangered Cathaya argyrophylla (Pinaceae) inferred from sequence variation of mitochondrial and nuclear DNA.

Cathaya argyrophylla is an endangered conifer restricted to subtropical mountains of China. To study phylogeographical pattern and demographic history of C. argyrophylla, species-wide genetic variation was investigated using sequences of maternally inherited mtDNA and biparentally inherited nuclear DNA. Of 15 populations sampled from all four distinct regions, only three mitotypes were detected at two loci, without single region having a mixed composition (G(ST) = 1). Average nucleotide diversity (theta(ws) = 0.0024; pi(s) = 0.0029) across eight nuclear loci is significantly lower than those found for other conifers (theta(ws) = 0.003 approximately 0.015; pi(s) = 0.002 approximately 0.012) based on estimates of multiple loci. Because of its highest diversity among the eight nuclear loci and evolving neutrally, one locus (2009) was further used for phylogeographical studies and eight haplotypes resulting from 12 polymorphic sites were obtained from 98 individuals. All the four distinct regions had at least four haplotypes, with the Dalou region (DL) having the highest diversity and the Bamian region (BM) the lowest, paralleling the result of the eight nuclear loci. An AMOVA revealed significant proportion of diversity attributable to differences among regions (13.4%) and among populations within regions (8.9%). F(ST) analysis also indicated significantly high differentiation among populations (F(ST) = 0.22) and between regions (F(ST) = 0.12-0.38). Non-overlapping distribution of mitotypes and high genetic differentiation among the distinct geographical groups suggest the existence of at least four separate glacial refugia. Based on network and mismatch distribution analyses, we do not find evidence of long distance dispersal and population expansion in C. argyrophylla. Ex situ conservation and artificial crossing are recommended for the management of this endangered species.

Cell Nucleus↗

High levels of variation despite genetic fragmentation in populations of the endangered mountain pygmy-possum, Burramys parvus, in alpine Australia.

In endangered mammals, levels of genetic variation are often low and this is accompanied by genetic divergence among populations. The mountain pygmy-possum (Burramys parvus) is an endangered marsupial restricted to the alpine region of Victoria and New South Wales, Australia. By scoring variation at eight microsatellite loci, we found that B. parvus populations exhibit high levels of genetic divergence and fall into three distinct groups from the northern, central and southern areas of the distribution of this species, consistent with previous assessments of mitochondrial DNA variation. F(ST) values between populations from these regions ranged from 0.19 to 0.54. Within the central area, there was further genetic fragmentation, and a linear association between genetic and geographical distance. This pattern is likely to reflect limited dispersal across barriers despite the fact that individual B. parvus can move several kilometres. Levels of genetic variation within populations were high with the exception of a southern population where there was evidence of inbreeding. From a conservation perspective, all three areas where B. parvus are found should be considered as separate gene pools; management of populations within these areas needs to take into account the low gene flow between populations, as well as threats posed by roads, resorts and other developments in the alpine region. The low genetic variability and inbreeding in the southern population is of particular concern given the high levels of variability in other B. parvus populations.

Animals↗

Inbreeding and reproduction in endangered ungulates: preservation of genetic variation through the Organization of Genetic Resource Banks.

There is a constant increase in the number of species suffering marked reductions in population size. This reduction in size and the lack of genetic flow may lead to a decrease in genetic variability and to matings between close relatives (i.e. inbreeding) with an ensuing reduction in fitness. It is thus important to understand the mechanism underlying the deleterious effects of inbreeding and to develop reproductive biotechnologies that will allow the reduction of inbreeding depression by facilitating gene exchange between populations. The study of three endangered species of gazelles, Cuvier's gazelle (Gazella cuvieri), Mohor gazelle (Gazella dama mhorr) and dorcas gazelle (Gazella dorcas neglecta) has revealed that inbreeding negatively affects several semen parameters (motility, sperm morphology, acrosome integrity). Semen cryopreservation has been achieved in the three species but success varies depending on the diluent employed and the level of inbreeding. Artificial insemination of Mohor gazelles have led to the birth of the first gazelle born using frozen-thawed semen but improvements are needed before this technology can be applied on a routine basis for the genetic management of the populations. Collection of oocytes after ovarian stimulation, followed by in vitro maturation, fertilization and culture has met with some initial success in the Mohor gazelle. These, together with other reproductive technologies, will offer an invaluable help in preserving the maximum of genetic diversity of these and related endangered ungulate species.

Animals↗

Centennial decline in the trophic level of an endangered seabird after fisheries decline.

Coastal marine ecosystems worldwide have undergone such profound transformations from over-fishing that trophic interactions observed today might be artifacts of these changes. We determined whether the trophic level of an endangered seabird, the Marbled Murrelet (Brachyramphus marmoratus), has declined over the past 100 years after the collapse of Pacific sardine (Sardinops sadax) fisheries in the late 1940s and the recent declines of similar fisheries in central California. We compared stable-isotope signatures of delta15N and delta13C in feathers of museum specimens collected before fisheries decline with values in murrelet feathers collected recently. Values of delta15N in prebreeding diets declined significantly, 1.4 per thousand or 38% of a trophic level, over the past century during cool ocean conditions and by 0.5 per thousand during warm conditions, whereas postbreeding values of delta15N were nearly constant. The delta13C values in prebreeding diets declined by 0.8 per thousand, suggesting an increased importance of krill in modern compared with historic prebreeding diets, but postbreeding diets did not change. Stable-isotope mixing models indicated that the proportion of energetically superior high-trophic-level prey declined strongly whereas energetically poor low-trophic-level and midtrophic-level prey increased in the prebreeding diet in cool years when murrelet reproduction was likely to be high. Decreased prey resources have caused murrelets to fish further down on the food web, appear partly responsible for poor murrelet reproduction, and may have contributed to its listing under the U.S. Endangered Species Act.

Animals↗

Genetically engineered plants, endangered species, and risk: a temporal and spatial exposure assessment for Karner blue butterfly larvae and Bt maize pollen.

Genetically engineered maize (Zea mays) containing insecticidal endotoxin proteins from Bacillus thuringiensis (Bt) delta-endotoxin proteins has been adopted widely in the Midwestern United States. The proteins are toxic to several lepidopteran species and because a variety of maize tissues, including pollen, may express the endotoxins, the probability of exposure to nontarget species, including endangered species, needs to be understood. The objective of this study was to assess the potential temporal and spatial exposure of endangered Karner blue butterfly larvae (Lycaeides melissa samuelis) to Bt maize pollen in Wisconsin using probabilistic exposure techniques and geographic information systems analysis. Based on degree-day modeling of butterfly phenology and maize pollen shed, there is some potential for temporal exposure of larvae to maize pollen. However, in the majority of years and locations, maize pollen shed most likely will occur after the majority of larval feeding on wild lupine (Lupinus perennis). The spatial analysis indicates that some Karner blue butterfly populations occur in close proximity to maize fields, but in the vast majority of cases the butterfly's host plant and maize fields are separated by more than 500 m. A small number of potential or existing Karner blue butterfly sites are located near maize fields, including sites in two of the four counties where temporal overlap is most likely. The exposure assessment indicates that these two counties should receive the highest priority to determine if Karner blue butterfly larvae are actually at risk and then, if needed, to reduce or prevent exposure.

Animals↗

Potential of assisted breeding techniques for the conservation of endangered mammalian species in captivity: a review.

An alarming worldwide extinction of animal species is taking place as a result of the activities of the increasing global human population. The original ranges of many animal species are being reduced and fragmented and, in some cases, they have been reduced to perilously small relict populations. The adverse genetic consequences of these restrictions are becoming clear, as are possible methods for their alleviation. The concept of ex situ genetic management of small captive populations of endangered species with a view to re-introducing them into the wild is attracting increasing interest. Modern reproductive techniques will play an important role in such programmes, and it is likely that an increasing number of veterinarians will become involved. However, the literature describing the aims and methods of reproductive genetic management is scattered and often not readily available to interested veterinary surgeons. The aim of this review is to deal with this problem by describing some potential approaches to the captive breeding of endangered species.

Animal Husbandry↗