Artificial insemination for the propagation of CANDES: the reality!

Conservation is about protecting and nurturing species so that they can survive, not only now, but also into the future. Ideally this means protecting genetically diverse populations and not simply breeding a few individuals. Unfortunately, this point is often overlooked by reproductive technologists, especially if they are more accustomed to working with humans, companion animals or agricultural species, where the goals are more usually directed towards obtaining offspring from particular individuals. This approach has tended to antagonise the conservation community, who are quick to develop an unreasonable suspicion of technological solutions, partly because they are unfamiliar with the scientific principles that underpin the reproductive technology. Unfortunately, this mutual failure to recognise that all parties are actually well meaning, has led to separate cultures that barely communicate with each other and thus fail to capitalise on the potential benefits that would come from a good working relationship. Notable successes with reproductive technology have only emerged where such relationships have been forged. In this review, we highlight, mainly for the benefit of the technologist community, the need to foster good working relationships with conservation managers and to recognise that the latest hi-tech approach to animal breeding is more likely to engender suspicion than enthusiasm.     

Artificial insemination in deer and non-domestic bovids

Artificial insemination technology has revolutionized the domestic cattle breeding industry and allowed for the dissemination of valuable genetics worldwide. This technology has been adapted for use in many other taxa for the conservation of threatened and endangered species, but its use for the genetic management of small populations of deer, antelope and other non-domestic bovids has met numerous challenges and limited success. In practice, adaptation of domestic bovine AI protocols to other artiodactylids for genetic management has been limited by: (1) a lack of understanding of species-specific reproductive characteristics; (2) the inability to minimize handling stress; (3) pregnancy losses; and (4) regulatory challenges in semen importation. To date, AI protocols have been developed for seven species of cervid and 14 species of non-domestic bovids; recent developments in this technology has allowed greater use of AI for dissemination of genetics in farmed deer species. However, despite decades of research in the use of assisted reproduction for the conservation of antelope and other non-domestic bovids, even this simplest technique has not been used repeatedly for genetic management.     

Implementing artificial insemination as an effective tool for ex situ conservation of endangered avian species

Approximately 503 of the known species of birds are classified as ‘endangered’ or ‘critical’. Captive propagation programs have proven useful in maintaining genetic diversity and restoring wild populations of certain species, including the Peregrine falcon, California condor and Whooping crane. Artificial insemination (AI) has the potential of solving problems inherent to reproductive management of small, closed populations of endangered birds, including dealing with demographic instability, physical and behavioral disabilities, sexual incompatibility, lack of synchrony, and need to maintain gene diversity. In this review, we address the necessary methods and factors that allow AI to be applied effectively to manage rare bird populations. It is clear that semen availability and quality are the greatest limiting factors to implementing consistently successful AI for birds. Behavioral sensitivity to animal handling and the ability to minimize stress in individual birds also are keys to success. Multiple, deep vaginal inseminations can improve fertility, particularly when semen quality is marginal. Laparoscopic methods of semen transfer also have produced fertile eggs. All of these practices leading to successful AI remain dependent on having adequate basic knowledge on female reproductive status, copulatory behavior, endocrine profiles and duration of fertility, especially as related to oviposition. The overall greatest challenge and highest priority is defining these normative traits, which are highly species-specific.     

Comparison of conventional and computer-assisted semen analysis in cockatiels (Nymphicus hollandicus) and evaluation of different insemination dosages for artificial insemination

Many psittacine species are threatened in the wild and also rare in captivity. Therefore, successful conservation and breeding programs are important to save these species. Unfortunately, clutches in conservation programs are frequently infertile. Semen evaluation is beneficial to investigate the causes of infertility and is advisable before artificial insemination (AI). In this study, we analyzed the semen of cockatiels (Nymphicus hollandicus) using two different methods and investigated different insemination dosages for AI. Cockatiels (n = 30) were divided into two groups (group A: nine males; group B: six males). The males in group B were endoscopically sterilized, whereas the males in group A were used as semen donors. In the first part of the study, the semen of males in group A was evaluated by semen analysis. Semen samples were collected by the massage technique and examined using a conventional light microscope and a computer-assisted semen analyzer for comparison. Results demonstrated that the evaluations of motility, progressive motility, and sperm concentration, but not of live/dead ratio, correlated strongly for both methods. However, the results for sperm concentration, progressive motility, and live/dead ratio differed significantly. In the second part of our study, the volume and quantity of spermatozoa of the semen samples were adjusted and used for AI of females of group B. Intravaginal insemination with 250,000 spermatozoa resulted in five of 17 (29%) eggs fertilized; however, intracloacal insemination resulted in only four of 57 (7%) eggs fertilized at 232,000 and 250,000 spermatozoa but none at higher or lower dosages.     

Chemical constituents of the Korean endangered species Rhododendron brachycarpum

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The impact of climate change and potential distribution of the endangered white winged wood duck (Asarcornis scutulata, 1882) in Indian eastern Himalaya

The White-Winged Wood duck (Asarcornis scutulata) is an endangered forest wetland bird currently on the verge of extinction due to an array of anthropogenic pressures. It has been reported that global climate change could affect the distribution of many bird species globally. Therefore, an understanding the potential distribution of the White-Winged Wood duck in future climate scenarios could facilitate the creation of immediate conservation plans and the mitigation of subsequent threats. This is the first ever study on the distribution of White-Winged Wood Duck (WWWD) where Representative Concentration Pathway (RCP) 8.5 scenario was used to forecast the distribution of the WWWD in the Indian Eastern Himalayan region in the 2050s and 2070s. The study revealed that 1.87 % of the total area of IEH has the high potential distribution of WWWD. The state of Assam alone includes 1.68 % of the highly potential habitat in the region. It was predicted that 436.61 km² of highly potential habitat would be lost by 2070. Changes in the annual temperature range, precipitation in the wettest months (June to September), and precipitation decrease in the warmest quarter (October to December) would result in the loss of highly potential habitats. Under the influence of climate change, the habitat of WWWD in the eastern part of the region is likely to shift towards the western part. It was found that there will be a decline in potential habitat in the Indian states of Arunachal Pradesh, Assam, Nagaland, and Tripura located in the IEH under future climate scenarios. The potential of areas located at the Bhutan and Assam border would increase for supporting WWWD as this species' requires the average annual precipitation about 1000–1200 mm. However, the simultaneous anthropogenic activity would further destroy potential habitats in the future. The current study has provided baseline data on the potential distribution of WWWD in the IEH region for immediate conservation management plans.     

Genetic diversity of two endemic and endangered Plantago species

Plantago algarbiensis and Plantago almogravensis are two endangered and endemic species from Portugal. Due to the rarity and endangered nature of these species as well as the lack of molecular data, their genetic variation was evaluated using ISSR and RAPD markers. P. algarbiensis species showed higher genetic variability (73.9% of polymorphism) than P. almogravensis (61.2%). The two species revealed a high level of genetic diversity, with a Nei's genetic diversity of 0.1965 and 0.2309 and a Shannon's diversity index of 0.2975 and 0.3520, for P. almogravensis and P. algarbiensis, respectively. A low level of genetic differentiation was observed (Gst = 0.1873) among the species. However, the cluster and PCA analyses, based on genetic similarity, revealed two main, clearly separate clusters, which directly corresponded to the plants isolated from each species. In situ and ex situ measures should be applied in order to preserve both species but, based on these results, P. almogravensis population should be a priority for conservation.     

Measuring progress in US endangered species conservation

Since passage of the Endangered Species Act in 1973, over 1300 endangered and threatened species have been protected in the USA and its territories. Most species continue to face a significant risk of extinction, but the status of many species is improving. Here we present analyses of federal agency reports to the United States Congress (1988–2002) that describe differences in species status and show which variables are correlated with improving or declining status. We found that 52% of species showed repeated improvements or were not declining over this time. Species status improves over time, with only 35% still declining 13 years or more after protection. Taxonomy, funding by US Fish and Wildlife Service and National Oceanic and Atmospheric Administration, and agency assessment of risk of extinction and potential to recover were significantly correlated with status.     

Molecular evidence on the species status and phylogenetic relationships of Spiranthes parksii, an endangered orchid from Texas

New genetic information on the taxonomic status of an endangered species may have strong implications for its continued protection and that of other listed taxa with questionable species status. While analyzing a more comprehensive project on the molecular phylogeny of North American Spiranthes (Orchidaceae), lack of genetic differentiation between endangered S. parksii, endemic only to Texas, and a common local congener, S. cernua, was discovered. To confirm these results in a comparative context, we focused parsimony analyses on a subset of data from local Spiranthes and the same congeners found elsewhere, expanded by additional local samples of the two focal species. Four DNA sequence segments (two chloroplast, one mitochondrial, one nuclear ribosomal) totaling 3191 base pairs were used separately and together to verify that S. parksii is nested within the same clade as S. cernua, and thus likely to be the same species. Our results concur with another recent investigation using AFLP and microsatellite data that also suggests S. parksii is not unique genetically. Therefore, based on empirical data and the phylogenetic species concept, endangered S. parksii is merely an aberrant local phenotype of and a synonym for S. cernua sensu lato. Our results should be considered during the current review of this orchid’s listing status, despite some recent policy changes on consideration of genetic data after listing.     

中国外来动物物种组成及跨境风险

[目的] 明确全球变化背景下我国外来动物物种的发生规律,进一步掌握我国生物入侵现状及发生动态。[方法] 利用公开数据库等系统收集和整理我国外来动物的物种组成、可利用性、原产地及国内分布地等信息。[结果] 截至2020年,我国已有外来动物844种,隶属8门22纲87目291科591属,脊索动物门、节肢动物门、软体动物门是我国外来动物物种组成数量最多的3大类群。我国外来动物中,有益物种的数量多于有害物种的数量,还有少部分中性物种。不同分类阶元外来动物的可利用性不同,脊索动物门中有益物种的数量更多,节肢动物门中有害物种的数量更多。我国外来动物主要来源于亚洲、北美洲、欧洲,外来动物的国内分布与区域经济发展水平及对外交流程度相关,东南沿海地区数量多,西北内陆地区数量少,与他国相邻的广西和云南也有数量较多的外来动物。2010—2020年间记录的典型外来动物也符合此分布模式。[结论] 我国外来动物数量众多,组成多样,表现出不同的可利用性,其来源及分布表现出明显的空间格局,在全球变化背景下,应警惕外来物种的跨境入侵风险。     

Population characteristics of endangered species Taxus chinensis var. mairei and its conservation strategy in Shanxi, China

The conservation of the endemic, endangered, and first-class protected tree species, Taxus chinensis var. mairei (Lem. & H. Lev.) W. C. Cheng & L. K. Fu, is urgent in China. To assist the protection of this important medicinal plant species, the biological and ecological characteristics of its populations were investigated and analyzed in Shanxi. The study area, southeastern Shanxi, is a special region suitable for this species growth and development. The floristic composition, life form spectrum, and species diversity of T. chinensis var. mairei forests reflected the features of both warm-temperate and subtropical regions. Size classes in DBH and static life table analysis showed that the population structure of T. chinensis var. mairei was of the increase-type. Plant density, survivorship, and mortality decreased with increase in age. Distribution patterns of individuals were basically clumped and saplings tended to be distributed randomly. Disturbance from collection of leaves, branches, and stems for medicine was an important factor affecting population structure, dynamics, and distribution. The sex ratio of female/male and the efficiency of sexual reproduction were very low. The main way of reproduction for this species is vegetative propagation. Based on these characteristics, some suggestions, such as establishing nature reserves, improving population structure by cultivation and transplantation, developing seed gardens, and plantations, etc., for the conservation management of T. chinensis var. mairei and its forests in Shanxi are discussed.     

Artificial insemination of captive European brown hares (Lepus europaeus PALLAS, 1778) with fresh and cryopreserved semen derived from free-ranging males

This study aimed to establish artificial insemination (AI) protocols to predictably initiate pregnancy during the breeding season in the European brown hare (EBH) (Lepus europaeus PALLAS, 1778). Semen was collected from seven captive and eight free-ranging males by means of electroejaculation. Semen from the free-ranging males was cryopreserved using directional freezing. Total motility/integrity of fresh and frozen-thawed semen was 91.6%/87.7% and 46.9%/53.8%, respectively. Ovulation was induced in ultrasonographically preselected females using a gonadotropin-releasing hormone analogue. Each female was inseminated with 1 mL fresh (Group A, n = 16) or frozen-thawed semen (Group B, n = 9) at a concentration of 100 × 10⁶ spermatozoa/mL. The use of ultrasonography (10 to 22 MHz) confirmed the intracervical semen deposit, the success of artificial ovulation induction (formation of postovulatory corpus luteum), and permitted the monitoring of individual pregnancies. Although sperm motility/integrity was significantly different between groups, no significant difference was detected in conception rates (A, 87.50%; B, 77.78%). Because of embryonic resorption, there was a slight difference in fertility rate between groups (A, 62.5%; B, 77.78%). Overall, AI in captive EBH using fresh and frozen-thawed semen achieved successful fertility rates. Long-term cryopreserved semen was used to bring new genetic material from the wild into a genetically limited captive population without extensive animal transport. Therefore, AI has the potential to enhance breeding programs for EBH especially when cryopreserved semen from wild donors is used.     

Characterization of male reproductive anatomy of the endangered Arabian oryx (Oryx leucoryx)

Reproductive tracts of four male Arabian oryx (Oryx leucoryx) from Shaumari Nature Reserve in Jordan were examined to characterize their reproductive anatomy. Animals were allocated into two groups based on their age: Group 1 (n = 2, males were 12 and 14 mo old) and Group 2 (n = 2, males were 7 and 9 yrs old). Observations regarding the morphology, position and orientation of different reproductive organs were made. The external and internal genital organs of male oryx were similar to other domestic ruminant species with minor differences.     

Multi-drug resistant,biofilm-producing high-risk clonal lineage of Klebsiella in companion and household animals

Antimicrobial resistance is a global emergency which needs one health approach to address. The present study was conducted to detect the prevalence of beta-lactamase and biofilm-producing Klebsiella strains in rectal swabs (n = 624) collected from healthy dogs, cats, sheep and goats reared as companion or household animals in India. The dogs and cats were frequently exposed to third- or fourth-generation cephalosporins for therapy. The sheep and goats were occasionally exposed to antibiotics and had environmental exposure. Phenotypical ESBL (n = 93) and ACBL (n = 88)-producing Klebsiella were isolated significantly more (P < 0·05) from companion animals than household animals. Majority of the Klebsiella possessed bla_CTX-M-15. The sequences bla_CTX-M-15.2, bla_CTX-M-197 and bla_CTX-M-225 are reported first time from the companion animals. All ACBL-producing isolates possessed bla_AmpC. The present study detected 65·8% of Klebsiella strains as biofilm producers possessing the studied biofilm associated genes. The isolates showed phenotypical resistance against chloramphenicol, tetracycline, doxycycline, co-trimoxazole, ampicillin, cefotaxime/clavulanic acid. The present study showed that companion and household animals (dogs, cats, sheep, goats) may act as a carrier of ESBL/biofilm-producing, multi-drug resistant, high-risk clonal lineage of Klebsiella.     

Microsatellite markers for the endangered Roanoke logperch, Percina rex (Percidae) and their potential utility for other darter species

The Roanoke logperch (Percina rex Jordan and Evermann), an endangered fish, occurs in only six watersheds in the Roanoke and Chowan river drainages of Virginia, USA. The species’ population genetic structure is poorly known. We developed 16 microsatellite markers that were reliably scorable and polymorphic P. rex. Markers were also screened in seven other darter species of the genus Percina. Most markers exhibited successful amplification and polymorphism in several species. These markers may therefore prove useful for population genetic studies in other darters, a diverse but highly imperiled group.     

Assessing the importance of artificial nest‐sites in the population dynamics of endangered Northern Aplomado Falcons Falco femoralis septentrionalis in South Texas using stochastic simulation models

Habitat availability might be the most important determinant of success for a species reintroduction programme, making investigation of the quality and quantity of habitat needed to produce self‐sustaining populations a research priority for reintroduction ecologists. We used a stochastic model of population dynamics to predict whether attempts to improve existing breeding territories using artificial nest platforms improved the population growth rate and persistence of a reintroduced population of Northern Aplomado Falcons Falco femoralis septentrionalis in South Texas. We further assessed whether the creation of new territories, i.e. conversion of entire areas to suitable habitat and not simply the erection of nest platforms, would lead to a subsequent increase in the nesting population. Our model was able to reproduce several characteristics of the wild population and predicted the number of breeding pairs per year strikingly well (R² = 0.97). Simulations revealed that the addition of nest platforms improved productivity such that the population would decline to extinction without them but is stable since their installation. Moreover, the model predicted that the increase in productivity due to nest platforms would cause the population to saturate available breeding territories, at which point the population would contain a moderate proportion of non‐territorial birds that could occupy territories if new ones become available. Population size would therefore be proportional to the increase in available territories. Our study demonstrates that artificial nest‐sites can be an effective tool for the management of reintroduced species.     

Spatio-temporal changes in habitat potential of endangered freshwater fish in Japan

In recent years, biodiversity conservation and ecosystem restoration have been key issues of watershed management in many countries. To maintain or restore the environmental quality of watersheds, we need to assess the impact of anthropogenic changes on stream ecosystems with accuracy. In addition, watershed conservation planners have to make strategic plans and determine priorities of each conservation activity.

A new monitoring methodology to evaluate the change of habitat condition for freshwater fish based on a predictive habitat model using logistic regression was developed and applied to the whole of Japan. The main contributions of our approach were 1) the construction of a Geographical Information System (GIS) database that integrates many types of data, including freshwater fish species, water quality, habitat fragmentation by damming, geology, and climate; 2) spatial analysis for quantitative assessment and predictive habitat modeling using logistic regression to combine fish survey data and environmental habitat factors to determine critical and major habitat variables for each target fish; and 3) digital mapping and changes detection of fish habitat potential for targeted endangered fish species to show habitat distribution and spatio-temporal changes of habitat potential over a 25-year period (from 1977 to 2002). We found that predicted suitable habitat and actual fish habitat showed high overlap, and that habitat conditions and distribution patterns of target freshwater fish had been affected by major habitat variables to target species respectively.