School of BioSciences - Research Publications
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ItemEthical Analysis of the Application of Assisted Reproduction Technologies in Biodiversity Conservation and the Case of White Rhinoceros (Ceratotherium simum) Ovum Pick-Up Procedures.(Frontiers Media SA, 2022)Originally applied on domestic and lab animals, assisted reproduction technologies (ARTs) have also found application in conservation breeding programs, where they can make the genetic management of populations more efficient, and increase the number of individuals per generation. However, their application in wildlife conservation opens up new ethical scenarios that have not yet been fully explored. This study presents a frame for the ethical analysis of the application of ART procedures in conservation based on the Ethical Matrix (EM), and discusses a specific case study-ovum pick-up (OPU) procedures performed in the current conservation efforts for the northern white rhinoceros (Ceratotherium simum cottoni)-providing a template for the assessment of ART procedures in projects involving other endangered species.
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ItemThe Mating Pattern of Captive Naked Mole-Rats Is Best Described by a Monogamy Model(Frontiers Media SA, 2022-05-11)Naked mole-rats form colonies with a single reproductively active female surrounded by subordinate workers. Workers perform offspring care, construction and defense of the burrow system, and food supply. Such division of labor, called “cooperative breeding,” is strongly associated with the evolution of monogamous mating behavior, as seen in several mammalian lineages. This association is explained by the evolutionary theory of kin selection, according to which a subordinate adult may help to raise other’s offspring if they are in full sibling relationship. In conflict with this theory, the naked mole-rat is widely considered to be polyandrous, based on reports on multiple males contributing to a colony’s progeny. In order to resolve this contrast, we undertook an in-depth microsatellite-based kinship analysis on captive colonies. Four independent colonies comprising a total of 265 animals were genotyped using a panel of 73 newly established microsatellite markers. Our results show that each mole-rat colony contains a single monogamous breeder pair, which translates to a reproductive skew of 100% for both sexes. This finding, also in conjunction with previously published parental data, favors monogamy as the best-fitting model to describe naked mole-rat reproduction patterns. Polyandry or other polygamous reproduction models are disfavored and should be considered as exceptional. Overall, the empirical genetic data are in agreement with the kin selection theory.
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ItemTrigeminal ganglion and sensory nerves suggest tactile specialization of elephants.(Elsevier BV, 2022-02-28)Sensory nerves are information bottlenecks giving rise to distinct sensory worlds across animal species.1 Here, we investigate trigeminal ganglion2,3 and sensory nerves4 of elephants. The elephant trigeminal ganglion is very large. Its maxillary branch, which gives rise to the infraorbital nerve innervating the trunk, has a larger diameter than the animal's spinal cord, i.e., trunk innervation is more substantive than connections of the brain to the rest of the body. Hundreds of satellite cells surround each trigeminal neuron, an indication of exceptional glial support to these large projection neurons.5-7 Fiber counts of Asian elephant infraorbital nerves of averaged 4,00,000 axons. The infraorbital nerve consists of axons that are ∼10 μm thick and it has a large diameter of 17 mm, roughly 3 times as thick as the optic and 6 times as thick as the vestibulocochlear nerve. In most mammals (including tactile specialists) optic nerve fibers8-10 greatly outnumber infraorbital nerve fibers,11,12 but in elephants the infraorbital nerve fiber count is only slightly lower than the optic nerve fiber count. Trunk innervation (nerves and ganglia) weighs ∼1.5 kg in elephant cows. Our findings characterize the elephant trigeminal ganglion as one of the largest known primary sensory structures and point to a high degree of tactile specialization in elephants.
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ItemLacunae rostralis: A new structure on the rostrum of sailfish Istiophorus platypterus.(Wiley, 2022-05)Recent comparative studies of billfishes (Istiophoridae and Xiphiidae) have provided evidence of differences in the form and function of the rostra (bill) among species. Here, we report the discovery of a new structure, lacuna rostralis, on the rostra of sailfish Istiophorus platypterus, which is absent on the rostra of swordfish Xiphias gladius, striped marlin Kajikia audax and blue marlin Makaira nigricans. The lacunae rostralis are small cavities that contain teeth. They were found on the ventral rostrum surface of all I. platypterus specimens examined and dorsally in half of them. Ventrally, the lacunae rostralis were most prominent in the mid-section of the rostrum. Dorsally, they occurred closer to the tip. The density of lacunae rostralis increased towards the rostrum tip but, because they are smaller in size, the percentage of rostrum coverage decreased. The teeth located within the lacunae rostralis were found to be different in size, location and orientation from the previously identified micro-teeth of billfish. We propose two potential functions of the lacunae rostralis that both relate to the use of the bill in feeding: mechanoreception of prey before tapping it with the bill and more efficient prey handling via the creation of suction, or physical grip.
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ItemDynamic three-dimensional computed tomographic imaging facilitates evaluation of the equine cervical articular process joint in motion.(Wiley, 2022-01-19)BACKGROUND: Dynamic computed tomography (CT) imaging has been introduced in human orthopaedics and is continuing to gain popularity. With dynamic CT, video sequences of anatomical structures can be evaluated in motion. OBJECTIVES: To investigate the feasibility of dynamic CT for diagnostic imaging of the equine cervical articular process joints (APJs) and to give a detailed description of the APJ movement pattern. STUDY DESIGN: Descriptive cadaver imaging. METHODS: Cervical specimens of twelve Warmblood horses were included. A custom-made motorised testing device was used to position and manipulate the neck specimens and perform dynamic 2D and 3D CT imaging. Images were obtained with a 320-detector-row CT scanner with a 160 mm wide-area (2D) solid-state detector design that allows image acquisition of a volumetric axial length of 160 mm without moving the CT couch. Dynamic videos were acquired and divided into four phases of movement. Three blinded observers used a subjective scale of 1 (excellent) to 4 (poor) to grade the overall image quality in each phases of motion cycle. RESULTS: With an overall median score of 1 the image quality, a significantly lower score was observed in the dynamic 3D videos over the four phases by the three observers compared with the 2D videos for both flexion (3D 95% CI: 1-2 and 2D 95% CI: 1-3; P = .007) and extension movement (3D 95% CI: 1-2 and 2D 95% CI: 1-3; P = .008). Median Translational displacement of the APJ surface was significantly greater in flexion than in extension movement (P = .002). MAIN LIMITATIONS: The small number of specimens included. Excision of spines and removal of musculature. CONCLUSIONS: The study is a first step in the investigation of the potential of dynamic 3D CT in veterinary medicine, a technique that has only begun to be explored and leaves much room for refinement prior to its introduction in routine practice. CT with a detector coverage of 16 cm and a rotation speed of 0.32 seconds provides high-quality images of moving objects and gives new insight into the movement pattern of equine cervical APJs.
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ItemCryopreservation of Giraffe Epidydimal Spermatozoa Using Different Extenders and Cryoprotectants(MDPI, 2022-04-01)Giraffe numbers have plummeted over the last 30 years by 30-40%. Thus, their conservation status has been raised from least concern to vulnerable. Efforts to manage in situ and ex situ populations are increasing. Assisted reproduction techniques (ART) such as sperm cryopreservation could help preserve the genetic diversity of giraffe subspecies and, when used for artificial inseminations, enhance genetic exchange between isolated populations. However, to date, the post-thaw motility of recovered sperm has been low and inconsistent. In this study, epididymal sperm collected from the testes of giraffes (n = 7) was frozen in three different extenders, namely, BotuCrio, Steridyl, and test egg yolk (TEY), each supplemented with one of two different cryoprotectants (5% glycerol or a mix of 1% glycerol and 4% methylformamide) and frozen over liquid nitrogen vapor. Across all three extenders, sperm showed significantly better post-thaw results when frozen with a mix of glycerol and methylformamide compared with glycerol alone. Sperm frozen with TEY and a mix of glycerol and methylformamide achieved superior post-thaw total and progressive sperm motility of 57 ± 3% and 45 ± 3%, respectively. These results show the benefit of using alternative cryoprotectants for freezing giraffe spermatozoa and could aid in the application of ARTs for giraffe subspecies or the closely related endangered Okapi.
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ItemNaïve-like pluripotency to pave the way for saving the northern white rhinoceros from extinction.(Springer Science and Business Media LLC, 2022-03-08)The northern white rhinoceros (NWR) is probably the earth's most endangered mammal. To rescue the functionally extinct species, we aim to employ induced pluripotent stem cells (iPSCs) to generate gametes and subsequently embryos in vitro. To elucidate the regulation of pluripotency and differentiation of NWR PSCs, we generated iPSCs from a deceased NWR female using episomal reprogramming, and observed surprising similarities to human PSCs. NWR iPSCs exhibit a broad differentiation potency into the three germ layers and trophoblast, and acquire a naïve-like state of pluripotency, which is pivotal to differentiate PSCs into primordial germ cells (PGCs). Naïve culturing conditions induced a similar expression profile of pluripotency related genes in NWR iPSCs and human ESCs. Furthermore, naïve-like NWR iPSCs displayed increased expression of naïve and PGC marker genes, and a higher integration propensity into developing mouse embryos. As the conversion process was aided by ectopic BCL2 expression, and we observed integration of reprogramming factors, the NWR iPSCs presented here are unsuitable for gamete production. However, the gained insights into the developmental potential of both primed and naïve-like NWR iPSCs are fundamental for in future PGC-specification in order to rescue the species from extinction using cryopreserved somatic cells.
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ItemAssessing Coagulation Parameters in Healthy Asian Elephants (Elephas maximus) from European and Thai Populations(MDPI, 2022-02-01)The Asian elephant population is continuously declining due to several extrinsic reasons in their range countries, but also due to diseases in captive populations worldwide. One of these diseases, the elephant endotheliotropic herpesvirus (EEHV) hemorrhagic disease, is very impactful because it particularly affects Asian elephant calves. It is commonly fatal and presents as an acute and generalized hemorrhagic syndrome. Therefore, having reference values of coagulation parameters, and obtaining such values for diseased animals in a very short time, is of great importance. We analyzed prothrombin time (PT), activated partial thromboplastin time (aPTT), and fibrinogen concentrations using a portable and fast point-of-care analyzer (VetScan Pro) in 127 Asian elephants from Thai camps and European captive herds. We found significantly different PT and aPTT coagulation times between elephants from the two regions, as well as clear differences in fibrinogen concentration. Nevertheless, these alterations were not expected to have biological or clinical implications. We have also sequenced the coagulation factor VII gene of 141 animals to assess the presence of a previously reported hereditary coagulation disorder in Asian elephants and to investigate the presence of other mutations. We did not find the previously reported mutation in our study population. Instead, we discovered the presence of several new single nucleotide polymorphisms, two of them being considered as deleterious by effect prediction software.
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ItemGenomic basis for skin phenotype and cold adaptation in the extinct Steller's sea cow.(American Association for the Advancement of Science (AAAS), 2022-02-04)Steller's sea cow, an extinct sirenian and one of the largest Quaternary mammals, was described by Georg Steller in 1741 and eradicated by humans within 27 years. Here, we complement Steller's descriptions with paleogenomic data from 12 individuals. We identified convergent evolution between Steller's sea cow and cetaceans but not extant sirenians, suggesting a role of several genes in adaptation to cold aquatic (or marine) environments. Among these are inactivations of lipoxygenase genes, which in humans and mouse models cause ichthyosis, a skin disease characterized by a thick, hyperkeratotic epidermis that recapitulates Steller's sea cows' reportedly bark-like skin. We also found that Steller's sea cows' abundance was continuously declining for tens of thousands of years before their description, implying that environmental changes also contributed to their extinction.
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ItemIncreasing the Yield and Cryosurvival of Spermatozoa from Rhinoceros Ejaculates Using the Enzyme Papain.(MDPI AG, 2022-01-18)The preservation of rhinoceros semen is vital for captive breeding programs. While successful collection and cryopreservation of rhinoceros semen has been reported, the volume and quality of semen produced is often low due to the high viscosity associated with ejaculates collected via electroejaculation. Reducing semen viscosity would enable access to previously unusable spermatozoa from viscous fractions and could improve quality post-thaw. The enzyme papain successfully reduced the viscosity of camelid semen but has yet to be tested in wildlife species. This study assessed the influence of papain on the in vitro quality of rhinoceros spermatozoa during cryopreservation using advanced semen assessment. In experiment 1, the motility of spermatozoa from the viscous fraction of an ejaculate, either untreated or treated with papain and its inhibitor E-64 prior to cryopreservation, was assessed post-thaw. In experiment 2, spermatozoa from papain-treated viscous fractions were compared to spermatozoa frozen from untreated sperm-rich fractions pre-freeze, as well as after 0, 1.5 and 3 h of incubation post-thaw (37 °C). Papain significantly increased the quantity of spermatozoa collected from ejaculates, as well as the motility prior to freezing. Papain also improved the post-thaw motility, velocity, linearity and straightness of samples compared to sperm-rich samples, with no detriment to sperm viability, lipid membrane disorder, production of ROS or DNA integrity (p < 0.05). Results show the benefit of supplementing rhinoceros spermatozoa with papain prior to cryopreservation on sperm cryosurvival and demonstrates the potential of using papain to improve the success of cryopreservation protocols, not only for the rhinoceros, but also for other wildlife species.