In vitro fertilization program in white rhinoceros.

In brief: To save endangered rhinoceros species, assisted reproductive technologies are warranted. We here report in vitro blastocyst generation of the Near-Threatened Southern white rhinoceros and, for the first time, also of the technically Extinct Northern white rhinoceros. Abstract: The Anthropocene is marked by a dramatic biodiversity decline, particularly affecting the family Rhinocerotidae. Three of five extant species are listed as Critically Endangered (Sumatran, Javan, black rhinoceros), one as Vulnerable (Indian rhinoceros), and only one white rhino (WR) subspecies, the Southern white rhinoceros (SWR), after more than a century of successful protection is currently classified as Near Threatened by the IUCN, while numbers again are declining. Conversely, in 2008, the SWR's northern counterpart and second WR subspecies, the Northern white rhinoceros (NWR), was declared extinct in the wild. Safeguarding these vanishing keystone species urgently requires new reproductive strategies. We here assess one such strategy, the novel in vitro fertilization program in SWR and - for the first-time NWR - regarding health effects, donor-related, and procedural factors. Over the past 8 years, we performed 65 procedures in 22 white rhinoceros females (20 SWR and 2 NWR) comprising hormonal ovarian stimulation, ovum pick-up (OPU), in vitro oocyte maturation, fertilization, embryo culture, and blastocyst cryopreservation, at an efficiency of 1.0 ± 1.3 blastocysts per OPU, generating 22 NWR, 19 SWR and 10 SWR/NWR hybrid blastocysts for the future generation of live offspring.

Ethical reasoning and participatory approach towards achieving regulatory processes for animal-visitor interactions (AVIs) in South Africa.

South Africa's wide range of animal facilities offers many different types of Animal-Visitor Interactions, wild animal encounters where animals and visitors come closer than in normal circumstances. The aim of this study was to provide a map of the ethically relevant aspects involved in AVIs in South Africa as a first step towards regulating these activities. A participative approach based on the ethical matrix, a tool which organizes the ethical standings of the stakeholders by three bearing ethical principles (wellbeing, autonomy, fairness), was applied. The matrix was populated through a top-down approach and refined by engaging stakeholders in a workshop and two online self-administrated surveys. The outcome is a map of the value demands concerning Animal Visitor Interactions. This map shows how the ethical acceptability of AVIs is linked to different relevant issues like animal welfare, education, biodiversity conservation, sustainability, human competency, facility mission, impact on scientific research and socio-economic outcomes. In addition, results highlighted the importance of cooperation among stakeholders and suggested that attention for animal welfare can inform decision making and inspire a multidisciplinary approach in implementing a regulatory frame for South African wildlife facilities.

Application of decision tools to ethical analysis in biodiversity conservation.

Achieving ethically responsible decisions is crucial for the success of biodiversity conservation projects. We adapted the ethical matrix, decision tree, and Bateson's cube to assist in the ethical analysis of complex conservation scenarios by structuring these tools so that they can implement the different value dimensions (environmental, social, and animal welfare) involved in conservation ethics. We then applied them to a case study relative to the decision-making process regarding whether or not to continue collecting biomaterial on the oldest of the two remaining northern white rhinoceroses (Ceratotherium simum cottoni), a functionally extinct subspecies of the white rhinoceros. We used the ethical matrix to gather ethical pros and cons and as a starting point for a participatory approach to ethical decision-making. We used decision trees to compare the different options at stake on the basis of a set of ethical desiderata. We used Bateson's cube to establish a threshold of ethical acceptability and model the results of a simple survey. The application of these tools proved to be pivotal in structuring the decision-making process and in helping reach a shared, reasoned, and transparent decision on the best option from an ethical point of view among those available.

Que se logren decisiones éticamente responsables es crucial para el éxito de los proyectos de conservación de la biodiversidad. Adaptamos la matriz ética, el árbol de decisión y el cubo de Bateson para apoyar con el análisis ético de escenarios de conservación compleja mediante la estructuración de estas herramientas de tal manera que puedan ejecutar las diferentes dimensiones de valor (ambiental, social y bienestar animal) involucradas en la ética de la conservación. Después aplicamos las herramientas a un estudio de caso relacionado con el proceso de toma de decisiones respecto a si se debe seguir o no recolectando material biológico del rinoceronte blanco del norte (Ceratotherium simum cottoni) más viejo (una subespecie funcionalmente extinta) de los dos que existen. Usamos la matriz ética como un punto de partida para una estrategia participativa para la toma ética de decisiones y para recopilar los pros y contras éticos. Usamos el árbol de decisión para comparar las diferentes opciones en juego con base en un conjunto de deseos éticos. Usamos el cubo de Bateson para establecer un umbral de aceptación ética y modelar los resultados de una encuesta simple. La aplicación de estas herramientas demostró ser central en la estructuración del proceso de toma de decisiones y en el apoyo para lograr una decisión compartida, razonada y transparente sobre la mejor opción a partir de un punto de vista ético entre aquellos disponibles.

Development of A Tool for Assessing the Reputation of Zoos: The Zoo Ethical Reputation Survey (ZERS).

Nowadays, most zoos have taken prominent and active positions in endangered species conservation and educating visitors about the value of biodiversity. However, to be effective and trusted in their mission, they must act ethically and have a good reputation. Yet, the drivers that can influence their reputation are still little investigated, and there are still few studies focused on assessing the reputation of these institutions. In the present work, we report the development of a tool, the Zoo Ethical Reputation Survey (ZERS), and its pilot application to assess the opinions of the visitors of two zoos, one in Italy and one in Germany, on drivers that may influence the ethical reputation of zoos. Preliminary results based on the answers of 274 respondents show that visitors' opinions on zoos acting with ethical responsibility are correlated with emotional appeal and familiarity with these institutions. The application of ZERS can help zoos identify weaknesses in their reputation and develop new strategies to improve people's attitudes towards them, bringing many benefits to the individual zoo and zoological institutions in general.

The Animal-Visitor Interaction Protocol (AVIP) for the assessment of Lemur catta walk-in enclosure in zoos.

Animal-Visitor Interactions (AVI) are activities offered by zoos and other tourism facilities, in which visitors come into close contact with animals. These activities can promote conservational and educational content, raise conservation mindedness and responsibility for the environment and animal welfare, but if not properly managed can jeopardize visitors' and animals' well-being and conservation efforts. The Animal-Visitor Interaction assessment Protocol (AVIP) has been designed to perform an integrated and multidisciplinary assessment of these activities, encompassing the "One Health, One Welfare" approach. AVIP throughout six different steps allows to assess the effects of AVIs both on animals, visitors, and the staff involved. Results can assist zoos to improve management decisions, ensure a transparent evaluation of their activities and promote conservation education goals. Lemurs walk-in enclosures have become increasingly popular among zoos, nevertheless studies focused on their assessment are still scarce. To validate AVIP to this particular AVI, we applied it to assess a walk-in enclosure hosting five Lemur catta in an Italian zoo. Results of behavioural and physiological analyses suggested no changes in animal welfare level and the Animal Welfare Risk Assessment showed low animal welfare risks. Two Visitor Experience Surveys were used to interview 291 visitors, showing that the assessed AVI could help promote the zoo's conservation objectives and visitor education. Risk Assessment found low and medium risks to the health and safety of visitors. Results were then combined to perform a final ethical assessment. Some potential ethical concerns were detected, but the outcomes indicated that these conflicts were well managed. In the context of recent findings AVIP demonstrated its potential for application also in assessing AVIs involving primates. Our findings confirmed the usefulness of AVIP in assessing and monitoring AVIs, allowing to gain key information in a single process on multiple welfare-related parameters, educational impact, safety of the main stakeholders involved, and ethical concerns.

Global response of conservationists across mass media likely constrained bat persecution due to COVID-19.

Most people lack direct experience with wildlife and form their risk perception primarily on information provided by the media. The way the media frames news may substantially shape public risk perception, promoting or discouraging public tolerance towards wildlife. At the onset of the COVID-19 pandemic, bats were suggested as the most plausible reservoir of the virus, and this became a recurrent topic in media reports, potentially strengthening a negative view of this ecologically important group. We investigated how media framed bats and bat-associated diseases before and during the COVID-19 pandemic by assessing the content of 2651 online reports published across 26 countries, to understand how and how quickly worldwide media may have affected the perception of bats. We show that the overabundance of poorly contextualized reports on bat-associated diseases likely increased the persecution towards bats immediately after the COVID-19 outbreak. However, the subsequent interventions of different conservation communication initiatives allowed pro-conservation messages to resonate across the global media, likely stemming an increase in bat persecution. Our results highlight the modus operandi of the global media regarding topical biodiversity issues, which has broad implications for species conservation. Knowing how the media acts is pivotal for anticipating the propagation of (mis)information and negative feelings towards wildlife. Working together with journalists by engaging in dialogue and exchanging experiences should be central in future conservation management.

Ethical Analysis of the Application of Assisted Reproduction Technologies in Biodiversity Conservation and the Case of White Rhinoceros (Ceratotherium simum) Ovum Pick-Up Procedures.

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.

The ART of bringing extinction to a freeze - History and future of species conservation, exemplified by rhinos.

The ongoing mass extinction of animal species at an unprecedented rate is largely caused by human activities. Progressive habitat destruction and fragmentation is resulting in accelerated loss of biodiversity on a global scale. Over decades, captive breeding programs of non-domestic species were characterized by efforts to optimize species-specific husbandry, to increase studbook-based animal exchange, and to improve enclosure designs. To counter the ongoing dramatic loss of biodiversity, new approaches are warranted. Recently, new ideas, particularly the application of assisted reproduction technologies (ART), have been incorporated into classical zoo breeding programs. These technologies include semen and oocyte collection, artificial insemination, and in-vitro embryo generation. More futuristic ideas of advanced ART (aART) implement recent advances in biotechnology and stem-cell related approaches such as cloning, inner cell mass transfer (ICM), and the stem-cell-associated techniques (SCAT) for the generation of gametes and ultimately embryos of highly endangered species, such as the northern white rhinoceros (Ceratotherium simum cottoni) of which only two female individuals are left. Both, ART and aART greatly depend on and benefit from the rapidly evolving cryopreservation techniques and biobanking not only of genetic, but also of viable cellular materials suitable for the generation of induced pluripotent stem cells (iPSC). The availability of cryopreserved materials bridges gaps in time and space, thereby optimizing the available genetic variability and enhancing the chance to restore viable populations.

Emotional States of African Elephants (Loxodonta africana) Kept for Animal-Visitor Interactions, as Perceived by People Differing in Age and Knowledge of the Species.

This study aimed to investigate how three groups of people of differing ages, and with differing knowledge of the species, perceived the emotional state of African elephants (Loxodonta africana) managed in captive and semi-captive environments. Fifteen video-clips of 18 elephants, observed during three different daily routines (release from and return to the night boma; interactions with visitors), were used for a free choice profiling assessment (FCP) and then analyzed with quantitative methods. A general Procrustes analysis identified two main descriptive dimensions of elephant behavioral expression explaining 27% and 19% of the variability in the children group, 19% and 23.7% in adults, and 21.8% and 17% in the expert group. All the descriptors the observers came up with showed a low level of correlation on the identified dimensions. All three observers' groups showed a degree of separation between captive and semi-captive management. Spearman analyses showed that stereotypic "trunk swirling" behavior correlated negatively with first dimension (free/friendly versus sad/bored) in the children's group; second dimension (agitated/confident versus angry/bored) amongst the adults; and first dimension (active/excited versus agitated/bored) amongst the experts. More studies are needed to investigate other potential differences in assessing elephants' emotional states by visitors of different ages and backgrounds.

An Ethical Assessment Tool (ETHAS) to Evaluate the Application of Assisted Reproductive Technologies in Mammals' Conservation: The Case of the Northern White Rhinoceros (Ceratotherium simum cottoni).

Assisted reproductive technologies (ARTs) can make a difference in biodiversity conservation. Their application, however, can create risks and raise ethical issues that need addressing. Unfortunately, there is a lack of attention to the topic in the scientific literature and, to our knowledge, there is no tool for the ethical assessment of ARTs in the context of conservation that has been described. This paper reports the first applications of the Ethical Assessment Tool (ETHAS) to trans-rectal ovum pick-up (OPU) and in vitro fertilization (IVF) procedures used in a northern white rhinoceros (Ceratotherium simum cottoni) conservation project. The ETHAS consists of two checklists, the Ethical Evaluation Sheet and the Ethical Risk Assessment, and is specifically customized for each ART procedure. It provides an integrated, multilevel and standardized self-assessment of the procedure under scrutiny, generating an ethical acceptability ranking (totally, partially, not acceptable) and a risk rank (low, medium, high), and, hence, allows for implementing measures to address or manage issues beforehand. The application of the ETHAS to the procedures performed on the northern white rhinoceros was effective in ensuring a high standard of procedures, contributing to the acceptability and improved communication among the project's partners. In turn, the tool itself was also refined through an iterative consultation process between experts and stakeholders.

Scientific and Ethical Issues in Exporting Welfare Findings to Different Animal Subpopulations: The Case of Semi-Captive Elephants Involved in Animal-Visitor Interactions (AVI) in South Africa.

Elephants are charismatic, cognitively highly-developed animals, whose management conditions can vary along a "wild-captive continuum." Several protocols have been proposed for the assessment of zoo elephants' welfare. It is important to investigate the possible limitations, if any, of extending findings from zoo elephants to conspecifics in a different dynamic in said "wild-captive continuum." In this paper, findings regarding two issues will be discussed: those regarding the external validity and those regarding the acceptability of management procedures as applied to semi-captive (i.e., able to roam freely for part of the day) elephants involved in visitor-interaction programs in South Africa. In a questionnaire-based survey, half of the responding experts stated that at least some of the welfare issues they ranked as the five most important in captive elephants' management had a different relevance for semi-captive individuals, resulting in 23.6% of the issues being rated differently. Moreover, there was no agreement among the experts on the ethical acceptability of any of the investigated procedures used in the management of semi-captive elephants involved in visitor-interaction programs. Caution is thus needed when exporting findings from one subpopulation of animals to another kept in different conditions and more scientific and ethical research is needed on the topic.

Assessing Animal Welfare in Animal-Visitor Interactions in Zoos and Other Facilities. A Pilot Study Involving Giraffes.

In recent years, awareness of the controversial aspects connected with wild animal-visitor interactions (AVIs) in zoos and other facilities has increased due to cultural changes. Therefore, the need to apply transparent procedures to evaluate AVIs programs in zoos and similar facilities has also increased. This study presents results of animal welfare's assessment of a pilot test of a protocol based on six steps that aim to explore and assess the overall value of AVIs considering the impact both on animals and visitors. In the present paper, we discuss the multifaceted approach to animal welfare assessment during animal-visitor interactions, combining quantitative behavioural observations/analysis and a welfare risk-assessment procedure, which forms the basis of the six-step protocol. Pilot testing of said approach to animal welfare assessment involved giraffes (Giraffa camelopardalis) in an Italian zoo. No change in behaviour, suggestive of an increased welfare risk to the animals, was found. The risk analysis reported overall low risks for welfare, whereas enclosure analysis highlighted that the enclosure was suitable for allowing interactions without jeopardising animal welfare, mainly because it allowed animals to choose whether to interact or withdraw from interactions without decreasing the space available to them.

Distances between individuals in an artificial herd of African elephants (Loxodonta africana africana) during resource utilisation in a semi-captive environment.

Space allowance and resource dispersion is recognised as an important factor affecting the welfare of elephants in captivity. In the present pilot study, we investigated distances kept among individuals in an artificially created semi-captive mixed-sex group of African elephants, when individuals were free to disperse. The study involved a herd of six elephants, three females (aged 11 to 16years), and three males (aged 15 to 23years). They were observed through instantaneous scan sampling in order to assess distances between individuals and body orientation in space and through continuous focal animal sampling to assess inter-specific social behaviour and general activity. A total of 312 suitable scans were collected for evaluation of distances between individuals. While foraging in absence of discernible space constraints, elephants maintained a distance equalling five or more body lengths in 63.9% of the scans, with wide differences between dyads. Little social behaviour, mainly affiliative, was recorded. The results of this pilot study suggest further scientific investigation could help to understand whether placing resources at five body lengths distance or over in a controlled environment could increase their simultaneous utilisation by all members of a group and contribute to decrease aggression. However, caution is warranted when applying results to different groups, environments and management regimes.