Journal of Zoological and Botanical Gardens

The species of the Bufonidae family exhibit a great diversity of habitats, diurnal or nocturnal habits, a complex evolutionary history, and a wide distribution, which makes this group suitable for morphological studies. In this work, we aimed to identify the existence of morphological patterns related to the habitat use and diurnal or nocturnal habits of Bufonidae in the Brazilian Amazon. To achieve this, we studied the morphological measurements of 210 specimens from three zoological collections and characterized the type of habitat and diurnality/nocturnality of the species. The morphological patterns and habitat use were investigated through principal component analysis (PCA) and multiple correspondence analysis (MCA), respectively. The evaluation of the relationships between morphological variation, habitat use, and diurnality/nocturnality was performed via redundancy analysis (RDA). Accordingly, Amazonian bufonids were divided into three morphological groups associated with different vegetation types and environments, demonstrating that body size is closely linked to diurnal or nocturnal life habits and habitat. Species with large body sizes are associated to anthropized areas, while intermediate and smaller species are associated with primary forests. Full article

Amphibian welfare, particularly pain assessment in anurans, remains understudied despite their ecological and biomedical significance. This study aimed to identify and validate operational pain indicators for adult anurans under professional care. A four-phase approach was used: a systematic literature review, expert validation with risk analysis, field validation in a zoological facility, and development of a preliminary pain index. From 158 publications, 16 potential indicators were identified, encompassing behavioural, clinical, and physiological signs. Expert evaluation by 28 professionals from 12 institutions refined this to seven indicators, achieving over 60% consensus: feeding behaviour changes, abnormal behaviour, impaired locomotion, oedema, reduced movement, retained skin post-moulting, and altered respiration. Field validation in 53 anurans confirmed high observability and ease of measurement, with feeding behaviour changes and oedema scoring highest for practicality (93.5% and 93.0%, respectively). These validated indicators provide a science-based foundation for routine welfare monitoring, enabling timely interventions. Their integration into husbandry protocols can enhance ethical standards, improve conservation outcomes, and increase public confidence in amphibian care, paving the way for a standardised anuran pain index. Full article

In recent years, animal welfare scientists working in professionally managed settings have increasingly focused on promoting resilience to enhance the quality of life of individual animals. Resilience—defined as an animal’s capacity to be minimally affected by a disturbance or to rapidly return to the physiological, behavioral, cognitive, health, affective, and production states that pertained before exposure to a disturbance—involves various systems and dynamic processes. There is evidence that resilience can be measured using a suite of species-specific indicators, including both behavioral measures and physiological biomarkers. These indicators should be tracked for individuals of the same species over time and across various conditions, events, and experiences. Large-scale, multi-institutional studies allow welfare scientists to collect cross-sectional data to identify “resilient phenotypes” for the species of interest. Ultimately, the focus should be on improving outcomes for individual animals as they face particular stressors, challenges, and environmental disturbances over their lifetime. Animal care specialists play a crucial role in helping animals build resilience by providing opportunities to engage in cognitive challenges, stimulating environments, and species-appropriate social interactions. This review defines resilience for animal welfare scientists, as well as discusses how to measure and promote resilience in animals residing in zoos and aquariums. Full article

Although long neglected, sound is now an increasing topic of interest in zoo and aquarium science. Research has examined the impact of sounds, from various sources, on zoo-housed species, noting that the influence of sound is varied and very context specific. The zoo’s sound environment is influenced by the animals, the built environment, vegetation, climatic, temporal and seasonal factors, equipment use, husbandry practices, and human presence. Different sounds can dominate an enclosure at certain times. This article discusses a workshop involving 12 zoo professionals, held in March 2020, that explored how sound is considered or overlooked in zoo animal management. Although insights are based on a small group, limiting generalisability, the findings highlight areas where further understanding is required and should encourage research extension to other groups of stakeholders. Delegates emphasised that the auditory needs of animals are often underappreciated and that the influence of sound depends on the species involved and how the sound may be perceived. Delegates highlighted the importance of species- and individual-specific approaches, predictability, and how animals have (any) control over sounds experienced in their enclosure. Routine operational sounds, such as closing gates or doors, may inadvertently stress animals, suggesting the need to consider sound in enclosure design and husbandry schedules. Outputs also stated that sound, when carefully managed, can act as enrichment through (for example) structured auditory cues or naturalistic sounds if ecologically relevant. Overall, our findings support integrating sound measurement into broader welfare assessment frameworks and enclosure planning, and they identify practical applications including sound mitigation, enrichment strategies, and staff training. Future research should include a wider range of species (especially understudied taxa), consider the experiences of a wider stakeholder demographic, and evaluate how sound is management in complex or high-traffic areas of the zoo. Full article

This study investigates the impact of a gamified educational experience on the training of future teachers, focusing on a visit to the Natural History Pavilion at the University of Almería, compared to a traditional instructional approach. A quasi-experimental pre-test–post-test design was employed with both undergraduate and master’s degree teacher education students, combining quantitative analysis with complementary data. The findings indicate that participants in the gamified groups demonstrated significant gains in knowledge acquisition following the visit, as well as higher levels of appreciation for the activity itself. This study concludes that gamification cognitively enriches the museum learning experience and stands out as an innovative and pedagogically relevant strategy in teacher education. Full article

Assessing the well-being of captive brown bears is essential for responsible species management and meeting European and national animal care standards outside their natural habitats. The present study aims to assess the welfare of captive brown bears in Bulgaria, considering environmental parameters, husbandry procedures, and the psychological and physical condition of the animals. The study was conducted on 28 brown bears in 11 zoos in Bulgaria and 19 bears in Bear Sanctuary Belitsa. A specific questionnaire and interviews were conducted with zoo managers and keepers. The welfare of the bears was evaluated based on their living conditions, husbandry, and physical and psychological well-being. The Nature and Animal Protection Center in Dobrich received the highest rating for the welfare of two brown bears among the zoos in the study. The study identified annual veterinary check-ups and enrichment programmes as areas needing improvement. Full article

Many zoos display animals in mixed-species exhibits where multiple different species share the same space and potentially interact. This study analyzes a mixed-species exhibit with three New World monkey species (white-faced saki, black-capped squirrel monkey, and common squirrel monkey) at the Buffalo Zoo to determine the interactions among species and how different species use the exhibit space differently. Data were collected over twelve months using scan sampling. The sakis were more likely to be in proximity (less than 1.5 m apart) with others than were the squirrel monkey species. The sakis spent 26% of the time in contact with another animal, while both squirrel monkey species spent less than 1% of the time in contact with another animal. The squirrel monkeys used significantly more of the exhibit space than the sakis. A small number of observations occurred when only the sakis were on exhibit, and while speculative at best, anecdotally the sakis used much more of the exhibit when the squirrel monkeys were not on exhibit. There are many compelling reasons for zoos to design mixed-species exhibits; however, consideration needs to be given to how mixed-species exhibits impact animal behavior. Full article

The capability to generate induced pluripotent stem cells (iPSCs) from adult somatic cells, enabling them to differentiate into any cell type, has been demonstrated in several studies. In humans and mice, iPSCs have been shown to differentiate into primordial germ cells (PGCs), spermatozoa, and oocytes. However, research on iPSCs in deer is novel. Despite the necessity for establishing germplasm banks from endangered cervid species, the collection and cryopreservation of gametes and embryos have proven complex for this group. Therefore, the focus of this study was to establish protocols for deriving stable iPSC lines from Blastocerus dichotomus (Marsh deer) using primary cells derived from antler, adipose tissue, or skin, with the ultimate goal of producing viable gametes in the future. To achieve this, two main reprogramming approaches were tested: (1) transfection using PiggyBac transposons (plasmid PB-TET-MKOS) delivered via electroporation and (2) lentiviral transduction using the STEMCCA system with either human (hOSKM) or murine (mOSKM) reprogramming factors. Both systems utilized murine embryonic fibroblasts (MEFs) as feeder cells. The PiggyBac system was further supplemented with a culture medium containing small molecules to aid reprogramming, including a GSK inhibitor, MEK inhibitor, ALK/TGF inhibitor, and thiazovivin. Initial colony formation was observed; however, these colonies failed to expand post-selection. Despite these challenges, important insights were gained that will inform and guide future studies toward the successful generation of iPSCs in deer. Full article

Extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae pose a significant public health risk. As zoos grow in popularity, exotic animals come into closer contact with humans, making them potential reservoirs of ESBLs. However, data on ESBL presence in Mexican zoos remains limited. For this reason, this study aimed to isolate and assess the antimicrobial susceptibility of Enterobacteriaceae that colonize avian species in a petting zoo and to identify any ESBL-producing isolates. Cloacal swabs were collected from 34 healthy birds at Miguel Angel de Quevedo Zoo, Veracruz, Mexico. Samples were analyzed microbiologically and molecularly to detect ESBL-encoding genes. A total of seventeen E. coli and one K. pneumoniae strains were isolated from cloacal swabs of bird species, and multidrug resistance (MDR) was found. The most frequently detected genes were bla_CTX-M-1 (16/18) and bla_TEM-1 (12/18). The detection of multidrug-resistant (MDR) strains carrying bla_CTX-M-1, bla_TEM-1, and bla_SHV genes highlights the potential role of birds as reservoirs and disseminators of antimicrobial resistance (AMR) in urban environments. To the best of our knowledge, this is the first study conducted in Mexico. In conclusion, MDR ESBL-producing bacteria were found in the fecal microbiota of bird species at a petting zoo in Mexico. The limitations of this study emphasize the need for a One Health approach to analyze the genome-wide isolates and epidemiology of antimicrobial-resistant bacteria in captive zoo animals in Mexico. This would support targeted surveillance efforts and help reduce the emergence and spread of resistant bacteria among zoo animals and visitors. Full article

This study explores the use of chlorography as a natural photographic developing technique that utilizes the decomposition of chlorophyll and other plant pigments through the action of sunlight. The developed images corresponded to previous research on changes in the iconography of the indigenous Salasaka people. In this context, this experimental project on natural photography is oriented toward the conservation of the ancestral knowledge of this community and the understanding of the native flora of Ecuador. We investigated the application of the contact image transfer technique with positive transparencies on leaves and flowers of 30 different species that grow in the Ecuadorian highlands, including leaves of vascular plants, as well as rose petals. The results showed that the clarity and contrast of chlorography depended on the plant species and exposure time. It was observed that fruit-bearing species produced more visible images than the leaves of other plants and rose petals, with species from the Passifloraceae family proving particularly effective. We interpreted these findings within the framework of plant photophysical mechanisms, proposing an inverse relationship between development efficiency and species’ non-photochemical quenching (NPQ) capacity. Furthermore, we interpreted the findings in relation to the photobleaching of pigments and compared chlorography with other natural photographic processes such as anthotypes. Key factors influencing the process were identified, such as the type of leaf, the intensity and duration of light, and the hydration of the plant material. It is concluded that chlorography is a viable, non-toxic, and environmentally friendly photographic alternative with potential applications in art, education, and research, although it presents challenges in terms of image permanence and reproducibility. Full article

An adult female bottlenose dolphin (Tursiops truncatus) housed at a public oceanarium presented with acute anorexia and lethargy. A blood analysis demonstrated mild leukocytosis, marked azotemia, hyperkalemia, and hyperphosphatemia suggestive of acute kidney injury or renal insufficiency. Ultrasound examination of the dolphin revealed ascites, pleural effusion, bilateral nephrolithiasis, mild hydronephrosis, and bilateral hydroureter consistent with bilateral post-renal obstruction. Initial treatment consisted of antibiotics, oral fluids, and anti-inflammatory treatment. Further imaging diagnosed bilateral obstructing ureteroliths at both ureteral orifice junctions of the urinary bladder. The dolphin’s azotemia and hyperkalemia were nonresponsive to traditional medical management; therefore, peritoneal dialysis was performed for emergent clinical stabilization. Peritoneal dialysis was conducted over 3 days and facilitated the patient to undergo laser lithotripsy of the offending ureteral obstruction. The dolphin made a full recovery following months of intensive medical treatment for complications from peritoneal dialysis and secondary peritonitis. This is the first documented case of successful, though complicated, peritoneal dialysis in a cetacean. Full article

Zoological environments aim to promote natural behaviours and optimal welfare conditions. Over the past decade, research on the use of artificial ultraviolet-B (UV-B) exposure has improved vitamin D₃ levels and reduced incidences of metabolic bone disease in diurnal primates; however, this has not been investigated in nocturnals. Aye-ayes (Daubentonia madagascariensis), nocturnal lemurs often housed indoors in zoos with little to no exposure to natural sunlight, have been reported to have low vitamin D₃ levels. This study aims to investigate the impacts of artificial UV-B as a supplemental healthcare strategy for aye-ayes, examining its influences on vitamin D₃ levels and positional sleeping behaviour. The 25-hydroxy-vitamin D₃ (25OHD₃) blood levels were tested before and after exposure to different levels of artificial UV-B and heat sources. Statistical analysis showed no correlation between UV-B and 25OHD₃ at group parameter levels. However, one individual showed a positive correlation. Sleeping position duration analysis showed a potential basking behaviour with the use of increased ear exposure and other thermoregulatory responses. Despite representing 8.06% of the European captive aye-aye population, these findings highlight the need for further research on vitamin D₃ parameters and responses to UV-B to optimise captive conditions and support the species’ long-term health. Full article

Environmental enrichment is essential for promoting species-specific behaviors and enhancing the welfare of zoo-housed animals. This study examined the behavioral responses of two juvenile male binturongs (Arctictis binturong) at the Minnesota Zoo to three enrichment stimuli: lavender oil (olfactory), thawed fish (olfactory and dietary), and hard-boiled egg (olfactory and dietary). Their behaviors were recorded using scan sampling before and after enrichment exposure, focusing on locomotion, foraging, resting, and visitor visibility. Food-based enrichments, particularly the hard-boiled egg, significantly increased foraging behavior, while lavender oil and thawed fish produced minimal behavioral changes. Locomotion and visibility remained stable across the conditions, although a slight increase in resting was observed with lavender oil. No evidence of scent-marking disruption was noted, and individual differences appeared to influence inactivity levels. These findings highlight the potential of biologically relevant, food-based enrichment to stimulate natural behaviors in binturongs and emphasize the importance of species-specific enrichment strategies. Future research should explore a broader range of olfactory cues, assess long-term behavioral responses, and incorporate physiological measures to further evaluate enrichment impacts on binturong welfare. Full article

Botanical gardens serve as vital centers for ex situ conservation, maintaining diverse plant species under controlled conditions. Terrestrial orchids, despite their wide diversity and distribution, often occur in small and declining populations, making their conservation increasingly urgent. This study aimed to examine the potential for establishing the first specialized orchidarium in Serbia, focusing on the native orchid species of the Fruška Gora region. A SWOT analysis, combined with site assessment data, was employed to identify key strengths, weaknesses, opportunities, and threats, informing the development of a functional zoning plan. The results indicate that such an orchidarium would offer a threefold benefit: strengthening ex situ conservation, advancing scientific research and environmental education, and promoting sustainable tourism. The proposed design consists of eight distinct zones, three of which reflect natural habitats of selected orchid species. The planned integration of a seed gene bank in the central zone, along with living plant collections and a nearby in vitro culture laboratory, establishes a comprehensive framework for the sustainable management of orchid genetic resources in the region, forming a foundation for future research and preservation. Full article

This contributed paper presents results from efforts by the Association of Zoos and Aquariums (AZA) to investigate the impact of integrating conservation into AZA members’ organizational cultures. Part of this work included AZA setting goals related to organizational and professional culture, strategic communication, developing communities of practice, and promoting tools and resource-sharing. Prior to implementing the majority of these steps, a baseline assessment was administered to directors plus a random sample of AZA organizations in 2020—assessing how well conservation is integrated into the institutional culture and measures of perceived conservation impact. The same sample of organizations was re-surveyed in 2023 with the intent of a three-year cycle of surveying to monitor change and identify additional ways that AZA could support and strengthen a culture of conservation within the profession. These findings will help the zoological and broader conservation community assess opportunities to integrate conservation into organizational cultures across a broad association for the purpose of achieving the mission and impact. Full article

Food-based environmental enrichment (EE) is a valuable strategy for stimulating foraging behaviour in fish under human care, as it increases the challenge of food acquisition and encourages prolonged engagement in this activity. Curimbas (Prochilodus argenteus) and pacus (Myleus micans) are fish species for which ex situ maintenance has become an important conservation measure. In this context, providing EE is essential to ensure high welfare standards. This study aimed to assess the effects of food enrichment on the behaviour of these two endemic species from the São Francisco River basin in Brazil. Behavioural data were collected across three experimental phases, including baseline, enrichment, and post-enrichment. Slow-dissolving food items known as “acorns” were introduced during the enrichment phase. Both species exhibited a marked reduction in inactivity throughout the study. For curimbas, the enrichment phase was associated with increased foraging, elevated agonistic interactions, and greater use of specific tank areas. Among pacus, inactivity significantly declined during the enrichment period. Additionally, the presence of visitors influenced behavioural patterns, promoting foraging activity while reducing inactivity and interactions with the enrichment device. These findings reinforce the value of incorporating environmental enrichment to promote the welfare of freshwater fish in public aquariums. Full article

In the present study, we examine the possibilities of planned generation in snakes by controlling the two most important factors for their estrus—ambient temperature and daylight hours. As a result of controlling these environmental parameters in an increasing gradient until reaching optimal values for the species, we observed copulations in late March and early April between a female Vipera ammodytes transcaucasiana and a male V. a. ammodytes. After three months of “pregnancy”, we obtained viable offspring in early July, which is about two months earlier compared to wild populations. The species used in the experimental setup, in natural conditions, usually produce offspring in late August to early October. Another aspect considered in the publication and followed in the experimental setup was to possibly test if in evolutionary and developmental aspect, both subspecies are closely related and interspecies breeding is possible (which might indicate mutual ancestry). The hybrid individuals were monitored during their entire development from newborns to subadults for pathological traits during development to roll-out crossbreeding incompatibility. In our pilot investigation, no acquired or inherited pathological traits have been observed. The individuals were consistent with feeding and exhibited excellent individual development. Future research coupled with genetic investigation can give valuable insight in the field, whether it is valid to regard the genera as a Vipera ammodytes complex or as different subspecies groups. Full article

Marine animals without faces (MAWFs), are some of the most important creatures maintaining the ecological balance in marine environments. How these animals are depicted across conservation organizations may impact public perceptions and conservation efforts. We assessed the online presentation of sea stars, jellies, and corals among all public websites of institutions accredited by the Association of Zoos and Aquariums (AZA) (N = 237). Among the organizations with an aquarium (n = 125), only 55 (44 percent) profiled at least one of the three animals, resulting in 89 total profiles. Five general approaches to characterizing these animals emerged: (1) scientific social distancing, (2) beautiful and eye-catching, (3) grotesque, otherworldly, and strange, (4) brainless beauties, and (5) objects of touch, entertainment, and experience. While some practices, like touch exhibits, can support empathy outcomes among visitors, online profile practices may contribute to the objectification of these animals among visitors, which could ultimately impact conservation attitudes, intentions, and behaviors. Full article

The welfare of animals in zoos has come under increasing scrutiny as public awareness grows around the biological needs of captive species. It is also becoming clear that promoting positive welfare experiences upholds population management and conservation aims. This paper re-evaluates current welfare frameworks in zoological institutions, advocating for evidence-based practices, multi-dimensional welfare metrics, and greater emphasis on species-specific needs, as well as the importance of input-based approaches to assess the welfare of zoo animals. By evaluating the limitations of current welfare practices (e.g., a lack of species-specific assessment protocols or sound husbandry evidence to base measures on) and presenting potential areas for improvement, this paper identifies ways that sound baselines for meaningful zoo animal welfare outputs can be created. Although current welfare policies from large zoo membership organisations stipulate assessment of welfare outputs as key to improving animal welfare standards, such outputs can only be positive if inputs are species-specific and relevant to the animals being housed. Practices such as the use of environmental enrichment (for example) need to be further refined to ensure they provide meaningful outputs (for the individuals) from the inputs that create them. Understanding the animal’s needs to ensure that the goal of enrichment is clear benefits both the animal who is provided with the enrichment and the human caregivers as husbandry and management becomes easier. A focus on welfare outputs is commendable and (especially when considering emotional outputs) is indeed a gold standard to aim for, yet we must not lose sight of striving for improvements to housing, husbandry, and species-specific care. Without such fundamental support from correct inputs, outputs are unlikely to be truly (or meaningfully) positive. Therefore, consistent re-examination of inputs is required to make sure they uphold an individual’s attainment of good welfare. Full article