Strong Population Genetic Structure and Cryptic Diversity in the Florida Bonneted Bat (Eumops Floridanus)

Knowledge of genetic structure is essential for the long-term management and conservation of endangered species. We report the results from a genetic examination of the federally endangered Florida bonneted bat (Eumops floridanus) sampled from its range in southern Florida, USA. Bonneted bats are primarily found in four regions separated by approximately 100 to 250 kms, including three western natural areas (BW, PC, and CC) and one urban population on the east coast [Miami-Dade County (MD)]. We used 22 microsatellite loci and cytochrome b sequences to assess the extent of connectivity and levels of genetic diversity. Regional populations were highly differentiated (FST = 0.178) and model-based and multivariate analyses showed that MD was the most distinct among pairwise comparisons. Regional populations are small (i.e., Ne < 100) but demographically stable. Estimates of contemporary migration and historic gene flow suggest that regional populations do not frequently exchange migrants, but simulations suggest that the divergence among western regions is likely a result of recent genetic drift rather than long-term isolation. Significantly, mitochondrial DNA revealed that haplotypes from MD were similar or shared with those recognized as Eumops ferox from Cuba and Jamaica, and divergent (1.5%) from the remainder of bonneted bats in Florida. Our data support the management of each of the four populations as distinct population segments, and that BW, PC and CC combined are on an independent evolutionary trajectory from bats in MD. Critically, bonneted bats in Florida appear to harbor cryptic diversity that will require a reassessment of their taxonomy.

Vitrification preserves murine ovarian follicular cell transcriptome in a 3D encapsulated in vitro follicle growth system

Vitrification is a method for long-term biological sample cryopreservation without causing intra- and extra-cellular ice formation. We recently established a novel closed vitrification system to cryopreserve mouse ovarian follicles. Using the 3D alginate hydrogel encapsulated in vitro follicle growth (eIVFG) method, we demonstrated that compared to freshly-harvested follicles, vitrified follicles had normal follicle and oocyte reproductive outcomes. However, it is unknown whether vitrification preserves molecular signatures of folliculogenesis, which is the primary research focus in this study. Six fresh and six vitrified antral follicles grown from eIVFG were collected on day 8 for the whole single-follicle RNA sequencing. Principal component analysis (PCA) and Pearson’s correlation analysis revealed that vitrified follicles had similar transcriptomic profiles to fresh follicles. There were 35 differentially expressed genes between vitrified and fresh follicles, however, none of those genes have been shown to be critical to folliculogenesis and oogenesis. Meanwhile, gene ontology (GO) and KEGG pathway analysis revealed that no GO terms or signaling pathways were significantly enriched. Furthermore, the expression of genes essential for the gonadotropin-dependent folliculogenesis and oogenesis were comparable between vitrified and fresh follicles. Taken together, these results demonstrate that vitrification preserves follicular cell transcriptome and molecular signatures of gonadotropin-dependent folliculogenesis in the eIVFG system, providing a robust model for fertility preservation, conservation of endangered species, and also establishing a high-content ovarian follicle biobank for studying ovarian biology and female reproductive toxicology.

Interspecies Reconstructed Embryonic Cell Interaction between Campbell Hamster (Phodopus campbelli) and Mice (Mus musculus)

Interspecies embryo transfer is a seldom-used method to increase the successful conservation of endangered species. The study aimed to determine the potential development of interspecies reconstructed embryos. The present study used two animal models, Campbell hamsters (Phodopus campbelli) and mice (Mus musculus). The isolated inner cell mass (ICM) of hamster embryos were injected into the mice embryos. The embryos were transferred to the pseudopregnant mice using non-surgery embryos transfer methods. The fetuses were collected at day 13.5 of gestation for morphometric measurement and cytochrome b (Cyt b) analysis which used to determine the species of obtained fetuses. The results showed that the viability, pregnancy rate, and embryonic implantation ability of the interpecies reconstructed embryos did not differ significantly (p>0.05) compared to non-reconstructed embryos. Morphometric measurement showed that the crown-rump (CR) and the weight of fetuses in the reconstructed group were significantly higher than non-reconstructed group (p<0.05). According to Cytb analysis, the species of obtained fetuses were mice, while the population of hamster cells were found only in the blighted ovum (resorption). Therefore, it can be concluded that interspecies reconstructed embryos are able to implant. However, the population of mice cells are only found to develop.

Amendments suggested for zoo medical research strategies with focus on the D-A-CH region

Objective Scientific exploration of zoo animals is one of the main missions of modern zoos. As scientific achievements are best reflected within scientific literature, we screened appropriate literature search outcomes from the 5 highest ranked zoos in the D-A-CH region (Germany-Austria-Switzerland) in order to deduce suggestions for optimizations of future research strategies. Material and methods Literature search was done by entering ”Zoo Vienna”, “Tiergarten Schönbrunn”, “Zoo Berlin”, “Tiergarten Berlin”, “Tierpark Hellabrunn”, “Tierpark Hagenbeck”, and “Zoo Zurich” on PubMed and Scopus for the period 2000–2020. These 5 European zoos were chosen due to their broad public recognition and international importance. Inclusion criterion for the literature list was the description/mentioning of analysed animals or samples with a clear affiliation to the respective zoo in the materials and methods or acknowledgements sections. Search hits were then allocated to the following 7 areas: animal nutrition, biology, ethology, infectiology, reproduction, phylogenetics, and clinical medicine. Also, portions of higher animal taxa (and species, if appropriate) were recorded. Results and conclusions A total of 142 papers has been published. Mammals, especially large ones, were clearly over-represented in literature with 2 thirds of analysed papers dealing with them. Sauropsids (birds and reptiles) were treated in 28 % and non-amniotic vertebrates (amphibians and fish) as well as invertebrates in only 3 % each. This apportionment is in no relation to the species numbers of the respective higher animal taxa. The predominating research areas (covered by approximately 75 % of papers) were ethological studies, followed by papers on infectious diseases, and finally papers on biology with morphological, physiological, and molecular biological themes. Research on reproductive biology/medicine, which is considered to be of tremendous importance for the establishment of ex-situ populations and thus for the conservation of endangered species, has been covered by only 6 % of papers. Future research should more intensely keep an eye on that discipline, especially in non-mammalian vertebrates.

Larval cryopreservation as new management tool for threatened clam fisheries

Cryopreservation is the only reliable method for long-term storage of biological material that guarantees genetic stability. This technique can be extremely useful for the conservation of endangered species and restock natural populations for declining species. Many factors have negatively affected the populations of high economical value shellfish in Spain and, as a result, many are declining or threatened nowadays. This study was focused on early-life stages of Venerupis corrugata, Ruditapes decussatus and Ruditapes philippinarum to develop successful protocols to enhance the conservation effort and sustainable shellfishery resources. Firstly, common cryoprotecting agents (CPAs) were tested to select the suitable permeable CPA attending to toxicity. Cryopreservation success using different combinations of CPA solutions, increasing equilibrium times and larval stages was evaluated attending to survival and shell growth at 2 days post-thawing. Older clam development stages were more tolerant to CPA toxicity, being ethylene-glycol (EG) and Propylene-glycol (PG) the least toxic CPAs. CPA solution containing EG yielded the highest post-thawing survival rate and the increase of equilibration time was not beneficial for clam larvae. Cryopreservation of trochophores yielded around 50% survivorship, whereas over 80% of cryopreserved D-larvae were able to recover after thawing.

Rancang Bangun Alat Batik Kulit Kepala Harimau Barongan Reyog Ponorogo Menggunakan Arduino

Ponorogo identik dengan kesenian dan kerajinan reyog, salah satunya adalah topeng Barongan. Material utama topeng Barongan adalah kayu dadap sebagai kerangka utama dan kulit harimau sebagai pelapis luarnya. Perburuan harimau dilarang keras dalam rangka konservasi satwa langka, sehingga memaksa pengrajin topeng Barongan mencari alternatif lain, salah satunya menggunakan kulit sapi yang diberi pola khas harimau. Namun teknik yang digunakan masih manual sehingga memerlukan ketelitian, ketekunan dan waktu yang relatif lama. Maka dalam penelitian ini dibuat alat batik kulit kepala harimau Barongan Reyog Ponorogo. Gerakan alat batik kulit Barongan diatur oleh G-Code dan diinput melalui Software Grbl sender. Arduino berfungsi sebagai kontroler yang selanjutnya dapat menggerakkan motor Stepper sebagai penggerak sumbu dan Solenoid sebagai penekan penbrush untuk pewarnaan. Hasil penelitian ini menunjukkan alat batik Barongan Reyog Ponorogo mampu membuat pola harimau pada ukuran 30 x 35 cm dan memiliki kualitas yang hampir sama dengan teknik manual dengan waktu pewarnaan 8 menit. Ponorogo has been famous for its Reyog arts and crafts, one of them is the Barongan mask. The main materials for the Barongan mask are Dadap wood as the mainframe and tiger leather as the outer coating. Tiger hunting is strictly prohibited in the context of the conservation of endangered species. To force the Barongan mask craftsmen for other alternatives, one of the alternatives is to use cow leather which is drawn with a typical tiger pattern. However, the technique used is still manual, meanwhile, the manual technique requires a lot of precision, persistence, and time. So the research aim is to design and implement the tool for drawing the Barongan tiger's scalp on Reyog Ponorogo. The movement of the Barongan leather batik tool was regulated by G-Code and input via the Grbl sender Software then accepted by Arduino as a controller which can then move the Stepper motor as an axis drive and the Solenoid as a pen brush press for coloring. The results of this study indicate that the Barongan Reyog Ponorogo batik tool can make a tiger pattern at a size of 30 x 35 cm and almost has the same quality as the manual technique with a coloring time of 8 minutes.

Elephant Genomes Reveal Accelerated Evolution in Mechanisms Underlying Disease Defenses

Disease susceptibility and resistance are important factors for the conservation of endangered species, including elephants. We analyzed pathology data from 26 zoos and report that Asian elephants have increased neoplasia and malignancy prevalence compared with African bush elephants. This is consistent with observed higher susceptibility to tuberculosis and elephant endotheliotropic herpesvirus (EEHV) in Asian elephants. To investigate genetic mechanisms underlying disease resistance, including differential responses between species, among other elephant traits, we sequenced multiple elephant genomes. We report a draft assembly for an Asian elephant, and defined 862 and 1,017 conserved potential regulatory elements in Asian and African bush elephants, respectively. In the genomes of both elephant species, conserved elements were significantly enriched with genes differentially expressed between the species. In Asian elephants, these putative regulatory regions were involved in immunity pathways including tumor-necrosis factor, which plays an important role in EEHV response. Genomic sequences of African bush, forest, and Asian elephant genomes revealed extensive sequence conservation at TP53 retrogene loci across three species, which may be related to TP53 functionality in elephant cancer resistance. Positive selection scans revealed outlier genes related to additional elephant traits. Our study suggests that gene regulation plays an important role in the differential inflammatory response of Asian and African elephants, leading to increased infectious disease and cancer susceptibility in Asian elephants. These genomic discoveries can inform future functional and translational studies aimed at identifying effective treatment approaches for ill elephants, which may improve conservation.

Field testing an “acoustic lighthouse”: Combined acoustic and visual cues provide a multimodal solution that reduces avian collision risk with tall human-made structures

Billions of birds fatally collide with human-made structures each year. These mortalities have consequences for population viability and conservation of endangered species. This source of human-wildlife conflict also places constraints on various industries. Furthermore, with continued increases in urbanization, the incidence of collisions continues to increase. Efforts to reduce collisions have largely focused on making structures more visible to birds through visual stimuli but have shown limited success. We investigated the efficacy of a multimodal combination of acoustic signals with visual cues to reduce avian collisions with tall structures in open airspace. Previous work has demonstrated that a combination of acoustic and visual cues can decrease collision risk of birds in captive flight trials. Extending to field tests, we predicted that novel acoustic signals would combine with the visual cues of tall communication towers to reduce collision risk for birds. We broadcast two audible frequency ranges (4 to 6 and 6 to 8 kHz) in front of tall communication towers at locations in the Atlantic migratory flyway of Virginia during annual migration and observed birds’ flight trajectories around the towers. We recorded an overall 12–16% lower rate of general bird activity surrounding towers during sound treatment conditions, compared with control (no broadcast sound) conditions. Furthermore, in 145 tracked “at-risk” flights, birds reduced flight velocity and deflected flight trajectories to a greater extent when exposed to the acoustic stimuli near the towers. In particular, the 4 to 6 kHz stimulus produced the greater effect sizes, with birds altering flight direction earlier in their trajectories and at larger distances from the towers, perhaps indicating that frequency range is more clearly audible to flying birds. This “acoustic lighthouse” concept reduces the risk of collision for birds in the field and could be applied to reduce collision risk associated with many human-made structures, such as wind turbines and tall buildings.